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Articles by Anastacia M. Garcia in JoVE

 JoVE Biology

Use of Time Lapse Microscopy to Visualize Anoxia-induced Suspended Animation in C. elegans Embryos

1Department of Biological Sciences, University of North Texas


JoVE 4319

Described here is an in vivo technique to image sub-cellular structures in animals exposed to anoxia using a gas flow through microincubation chamber in conjunction with a spinning disc confocal microscope. This method is straightforward and flexible enough to suit a variety of experimental parameters and model systems.

Other articles by Anastacia M. Garcia on PubMed

Aminopiperidine Sulfonamide Ca(v)2.2 Channel Inhibitors for the Treatment of Chronic Pain

The voltage-gated calcium channel Ca(v)2.2 (N-type calcium channel) is a critical regulator of synaptic transmission and has emerged as an attractive target for the treatment of chronic pain. We report here the discovery of sulfonamide-derived, state-dependent inhibitors of Ca(v)2.2. In particular, 19 is an inhibitor of Ca(v)2.2 that is selective over cardiac ion channels, with a good preclinical PK and biodistribution profile. This compound exhibits dose-dependent efficacy in preclinical models of inflammatory hyperalgesia and neuropathic allodynia and is devoid of ancillary cardiovascular or CNS pharmacology at the doses tested. Importantly, 19 exhibited no efficacy in Ca(v)2.2 gene-deleted mice. The discovery of metabolite 26 confounds further development of members of this aminopiperidine sulfonamide series. This discovery also suggests specific structural liabilities of this class of compounds that must be addressed.

Effects of Chronic Treatment with Corticosterone and Imipramine on Fos Immunoreactivity and Adult Hippocampal Neurogenesis

In a previous study we showed that rats chronically treated with corticosterone (CORT) display anxiogenic behavior, evidenced by facilitation of avoidance responses in the elevated T-maze (ETM) model of anxiety. Treatment with the tricyclic antidepressant imipramine significantly reversed the anxiogenic effects of CORT, while inhibiting ETM escape, a response related to panic disorder. To better understand the neurobiological mechanisms underlying these behavioral effects, analysis of c-fos protein immunoreactivity (fos-ir) was used here to map areas activated by chronic CORT (200mg pellets, 21-day release) and imipramine (15mg/kg, IP) administration. We also evaluated the number of cells expressing the neurogenesis marker doublecortin (DCX) in the hippocampus and measured plasma CORT levels on the 21st day of treatment. Results showed that CORT increased fos-ir in the ventrolateral septum, medial amygdala and paraventricular hypothalamic nucleus and decreased fos-ir in the lateral periaqueductal gray. Imipramine, on the other hand, increased fos-ir in the medial amygdala and decreased fos-ir in the anterior hypothalamus. CORT also decreased the number of DCX-positive cells in the ventral and dorsal hippocampus, an effect antagonized by imipramine. CORT levels were significantly higher after treatment. These data suggest that the behavioral effects of CORT and imipramine are mediated through specific, at times overlapping, neuronal circuits, which might be of relevance to a better understanding of the physiopathology of generalized anxiety and panic disorder.

Safety Profile of Biological Intravenous Therapy in a Rheumatoid Arthritis Patients Cohort. Clinical Nursing Monitoring (Sebiol Study)

INTRODUCTION: The Biologics used in the management of rheumatoid arthritis (RA) in recent years, have comprehensively permitted to understand its security, as shown in registries such as BIOBADASER. The present manuscript represents an observational cohort study to describe the safety perinfusional profile of those intravenous treatments. OBJECTIVES: To confirm the safety profile of biological therapies in routine clinical practice, after the administration of intravenous drugs and 24hours post-administration. MATERIAL AND METHODS: We evaluated a cross-sectional cohort of 114 patients with RA (according to the American College of Rheumatology ACR criteria), attending within one month in 2009 the nursing clinics of day care hospital of 12 Catalonian hospitals. All patients were treated with intravenous biological agents. We recorded the age, sex, current and previous drug treatments, we also collected data about previous vaccination and premedication received and any adverse event occurring at the time of drug administration or within 24hours. If an adverse event occurred, was categorized by MedDRAv11.0 International Dictionary, and categorized in terms of intensity (mild, moderate, severe), relationship to drug administration according to Karch and Lasagna algorithm (unrelated, unlikely, possible, probable, definite) and the further measures taken. RESULTS: 111 patients met the inclusion criteria, with a mean age of 56.06 years (SD: 12.12), 90 of them women (81.1%) and mean time since diagnosis of the disease of 11.97 years (SD: 7.95). 24 patients (21.6%) had a history of allergy. 12 adverse events were observed in 7 patients, 9 of which at the time of administration and 3 in 24hours after. There were no serious adverse events and only one of the adverse events (AEs) was rated as moderate (urticaria). The remaining AA were mild.

Pharmacokinetic Study of Growth Hormone-Releasing Peptide 6 (GHRP-6) in Nine Male Healthy Volunteers

GHRP-6 is a growth hormone secretagogue that also enhances tissue viability in different organs. In the present work, we studied the pharmacokinetics of this short therapeutic hexapeptide (His-(D-Trp)-Ala-Trp-(D-Phe)-Lys-NH(2,) MW=872.44Da) in nine male healthy volunteers after a single intravenous bolus administration of 100, 200 and 400μg/kg of body weight. GHRP-6 was quantified in human plasma by a specific LC-MS method, previously developed and validated following FDA guidelines, using (13)C(3)Ala-GHRP-6 as internal standard (Gil et al., 2012, J. Pharm. Biomed. Anal. 60, 19-25). The Lower Limit of Quantification (5ng/mL) was reached in all subjects at 12h post-administration, which was sufficient for modeling a pharmacokinetic profile including over 85% of the Area under the Curve (AUC). Disposition of GHRP-6 best fitted a bi-exponential function with R(2) higher than 0.99, according to a mathematic modeling and confirmed by an Akaike index (AIC) lower than that of the corresponding one-compartment model for all subjects. Averaging all three dose levels, the distribution and elimination half-life of GHRP-6 were 7.6±1.9min and 2.5±1.1h, respectively. These values are coherent with existing data for other drugs whose disposition also fits this model. Dose dependence analysis revealed a noticeable trend for AUC to increase proportionally with administered dose. Atypical GHRP-6 concentration spikes were observed during the elimination phase in four out of the nine subjects studied.

Phialemoniopsis, a New Genus of Sordariomycetes, and New Species of Phialemonium and Lecythophora

In molecular studies involving numerous clinical isolates of the genera Acremonium, Phialemonium and Lecytophora some of them could not be identified. To clarify the phylogenetic relationships among these fungi and other related taxa, we performed a polyphasic study based on a detailed morphological study and on the analysis of sequences of four loci: the internal transcribed spacer regions, the D1/D2 domains of the 28S rRNA, actin and β-tubulin genes. The combination of the resulting data let us propose the new genus Phialemoniopsis to accommodate the opportunistic fungi Phialemonium curvatum and Sarcopodium oculorum and two new species, Phialemoniopsis cornearis and Phialemoniopsis pluriloculosa. The taxonomy of Phialemoniopsis has not been completely resolved, however, remaining incertae sedis within the Sordariomycetes. In addition, the new species Lecythophora luteorubra, Lecythophora cateniformis and Phialemonium globosum are described and the species Acremonium atrogriseum and Taifanglania inflata are transferred to the genus Phialemonium. Lecythophora and Phialemonium are currently monophyletic genera of the families Coniochaetaceae (Coniochaetales) and Cephalothecaceae (Sordariales) respectively, according to our results. Tables summarizing key morphological features to distinguish the current species of Lecythophora, Phialemonium and Phialemoniopsis are provided.

Evaluation of Four Agrobacterium Tumefaciens Strains for the Genetic Transformation of Tomato (Solanum Lycopersicum L.) Cultivar Micro-Tom

KEY MESSAGE : Agrobacterium tumefaciens strains differ not only in their ability to transform tomato Micro-Tom, but also in the number of transgene copies that the strains integrate in the genome. The transformation efficiency of tomato (Solanum lycopersicum L.) cv. Micro-Tom with Agrobacterium tumefaciens strains AGL1, EHA105, GV3101, and MP90, harboring the plasmid pBI121 was compared. The presence of the nptII and/or uidA transgenes in regenerated T(0) plants was determined by PCR, Southern blotting, and/or GUS histochemical analyses. In addition, a rapid and reliable duplex, qPCR TaqMan assay was standardized to estimate transgene copy number. The highest transformation rate (65 %) was obtained with the Agrobacterium strain GV3101, followed by EHA105 (40 %), AGL1 (35 %), and MP90 (15 %). The mortality rate of cotyledons due to Agrobacterium overgrowth was the lowest with the strain GV3101. The Agrobacterium strain EHA105 was more efficient than GV3101 in the transfer of single T-DNA insertions of nptII and uidA transgenes into the tomato genome. Even though Agrobacterium strain MP90 had the lowest transformation rate of 15 %, the qPCR analysis showed that the strain MP90 was the most efficient in the transfer of single transgene insertions, and none of the transgenic plants produced with this strain had more than two insertion events in their genome. The combination of higher transformation efficiency and fewer transgene insertions in plants transformed using EHA105 makes this Agrobacterium strain optimal for functional genomics and biotechnological applications in tomato.

Field-scale Evaluation of Water Fluxes and Manure Solution Leaching in Feedlot Pen Soils

Accumulation of beef cattle manure on feedlot pen surfaces generates large amounts of dissolved solutes that can be mobilized by water fluxes, affecting surface and groundwater quality. Our objective was to examine the long-term impacts of a beef cattle feeding operation on water fluxes and manure leaching in feedlot pens located on sandy loam soils of the subhumid Sandy Pampa region in Argentina. Bulk density, gravimetric moisture content, and chloride concentration were quantified. Rain simulation trials were performed to estimate infiltration and runoff rates. Using chloride ion as a tracer, profile analysis techniques were applied to estimate the soil moisture flux and manure conservative chemical components leaching rates. An organic stratum was found over the surface of the pen soil, separated from the underlying soil by a highly compacted thin layer (the manure-soil interface). The soil beneath the organic layer showed greater bulk density in the A horizon than in the control soil and had greater moisture content. Greater concentrations of chloride were found as a consequence of the partial sealing of the manure-soil interface. Surface runoff was the dominant process in the feedlot pen soil, whereas infiltration was the main process in control soil. Soil moisture flux beneath pens decreased substantially after 15 yr of activity. The estimated minimum leaching rate of chloride was 13 times faster than the estimated soil moisture flux. This difference suggests that chloride ions are not exclusively transported by advective flow under our conditions but also by solute diffusion and preferential flow.

Novel DNA Damage Checkpoints Mediating Cell Death Induced by the NEDD8-activating Enzyme Inhibitor MLN4924

MLN4924 is an investigational small molecule inhibitor of the NEDD8-activating enzyme (NAE) in Phase I clinical trials. NAE inhibition prevents the ubiquitination and proteasomal degradation of substrates for cullin-RING ubiquitin E3 ligases that support cancer pathophysiology, but the genetic determinants conferring sensitivity to NAE inhibition are unknown. To address this gap in knowledge, we conducted a genome-wide siRNA screen to identify genes and pathways that affect the lethality of MLN4924 in melanoma cells. Of the 154 genes identified, approximately one-half interfered with components of the cell cycle, apoptotic machinery, ubiquitin system and DNA damage response pathways. In particular, genes involved in DNA replication, p53, BRCA1/BRCA2, transcription-coupled repair and base excision repair appeared to be important for MLN4924 lethality. In contrast, genes within the G2/M checkpoint affected sensitivity to MLN4924 in colon cancer cells. Cell cycle analysis in melanoma cells by flow cytometry following RNAi-mediated silencing demonstrated that MLN4924 prevented the transition of cells from S to G2 phase after induction of re-replication stress. Our analysis suggested an important role for the p21-dependent intra-S-phase checkpoint and extensive rereplication, whereas the ATR-dependent intra-S-phase checkpoint appeared to play a less dominant role. Unexpectedly, induction of the p21-dependent intra-S-phase checkpoint appeared to be independent of both Cdt1 stabilization and ATR signaling. Collectively, these data enhance our understanding of the mechanisms by which inhibition of NEDD8-dependent ubiquitination causes cell death, informing clinical development of MLN4924.

Quantitative Measurement of Allergen-specific Immunoglobulin E Levels in Mass Units (ng/mL): an Interlaboratory Comparison

Relationship Between Hospital Surgical Volume, Lobectomy Rates, and Adverse Perioperative Events at US Epilepsy Centers

Object Epilepsy surgery remains significantly underutilized. The authors recently reported that the number of lobectomies for localized intractable epilepsy in the US has not changed despite the implementation of clear evidence-based guidelines 10 years ago supporting early referral for surgery. To better understand why epilepsy surgery continues to be underused, the authors' objective was to carefully examine hospital-related factors related to the following: 1) where patients are being admitted for the evaluation of epilepsy, 2) rates of utilization for surgery across hospitals, and 3) perioperative morbidity between hospitals with low versus high volumes of epilepsy surgery. Methods The authors performed a population-based cohort study of US hospitals between 1990 and 2008 using the Nationwide Inpatient Sample (NIS), stratifying epilepsy surgery rates and trends as well as perioperative morbidity rates by hospital surgical volume. Results The number of lobectomies for epilepsy performed at high-volume centers (> 15 lobectomies/year) significantly decreased between 1990 and 2008 (F = 20.4, p < 0.001), while significantly more procedures were performed at middle-volume hospitals (5-15 lobectomies/year) over time (F = 16.1, p < 0.001). No time trend was observed for hospitals performing fewer than 5 procedures per year. However, patients admitted to high-volume centers were significantly more likely to receive lobectomy than those at low-volume hospitals (relative risk 1.05, 95% CI 1.03-1.08, p < 0.001). Also, the incidence of perioperative adverse events was significantly higher at low-volume hospitals (12.9%) than at high-volume centers (6.1%) (relative risk 1.08, 95% CI 1.03-1.07, p < 0.001). Conclusions Hospital volume is an important predictor of epilepsy surgery utilization and perioperative morbidity. Patients with medically refractory epilepsy should be referred to a comprehensive epilepsy treatment center for surgical evaluation by an experienced clinical team.

Precision Top-Quark Mass Measurement at CDF

We present a precision measurement of the top-quark mass using the full sample of Tevatron sqrt[s]=1.96  TeV proton-antiproton collisions collected by the CDF II detector, corresponding to an integrated luminosity of 8.7  fb^{-1}. Using a sample of tt[over ¯] candidate events decaying into the lepton+jets channel, we obtain distributions of the top-quark masses and the invariant mass of two jets from the W boson decays from data. We then compare these distributions to templates derived from signal and background samples to extract the top-quark mass and the energy scale of the calorimeter jets with in situ calibration. The likelihood fit of the templates from signal and background events to the data yields the single most-precise measurement of the top-quark mass, M_{top}=172.85±0.71(stat)±0.85(syst)  GeV/c^{2}.

Measurement of the Pseudorapidity and Centrality Dependence of the Transverse Energy Density in Pb-Pb Collisions at Sqrt[s_{NN}]=2.76  TeV

The transverse energy (E_{T}) in Pb-Pb collisions at 2.76 TeV nucleon-nucleon center-of-mass energy (sqrt[s_{NN}]) has been measured over a broad range of pseudorapidity (η) and collision centrality by using the CMS detector at the LHC. The transverse energy density per unit pseudorapidity (dE_{T}/dη) increases faster with collision energy than the charged particle multiplicity. This implies that the mean energy per particle is increasing with collision energy. At all pseudorapidities, the transverse energy per participating nucleon increases with the centrality of the collision. The ratio of transverse energy per unit pseudorapidity in peripheral to central collisions varies significantly as the pseudorapidity increases from η=0 to |η|=5.0. For the 5% most central collisions, the energy density per unit volume is estimated to be about 14  GeV/fm^{3} at a time of 1  fm/c after the collision. This is about 100 times larger than normal nuclear matter density and a factor of 2.6 times higher than the energy density reported at sqrt[s_{NN}]=200  GeV at the Relativistic Heavy Ion Collider.

Siderophores of Stenotrophomonas Maltophilia: Detection and Determination of Their Chemical Nature

Stenotrophomonas maltophilia is an emerging nosocomial pathogen. Despite the broad spectrum of syndromes associated with S. maltophilia infections, little is known about its virulence factors, including siderophore production. The aims of this work were to detect S. maltophilia siderophores and to determine their chemical nature. We studied 31 S. maltophilia isolates from device-associated infections, recovered over the period 2006-2011 at Hospital de Clínicas José de San Martín, Buenos Aires, Argentina, and the strain K279a, whose genome has been fully sequenced. The production of siderophores was screened by the chrome azurol S (CAS) agar assay, previously modified to detect siderophores in this species. When grown on modified CAS agar plates, all the clinical isolates and K279a were CAS-positive for siderophore production. In order to determine the chemical nature of siderophores, the Csáky (hydroxamate-type) and Arnow (catechol-type) assays were used. All S. maltophilia isolates produced catechol-type siderophores, but hydroxamate-type siderophores were not detected.

In Vivo and in Vitro Expression of the Plasminogen Activators and Urokinase Type Plasminogen Activator Receptor (u-PAR) in the Pig Oviduct

Plasminogen activator activities have previously been reported in oviductal fluid. At present the question was whether the source of these activities is molecules come from blood plasma or if these activators are synthesized by the oviduct. Gene expression and protein synthesis of urokinase type (u-PA) and tissue type (t-PA) occur in different regions of the pig oviduct. Their relative concentrations do not vary between the ampulla and isthmus regions and are similar throughout the estrous cycle. However, while relative amounts of t-PA mRNA were not different between the different stages of the estrous cycle, u-PA mRNA was greater after ovulation (P<0.05). Regarding the function of u-PA, its receptor (u-PAR) was distinguished by immunohistochemistry at the apical region of the epithelial cells and was more noticeable in the isthmus. Expression of u-PA, t-PA, u-PAR and PAI-1 genes in primary oviductal epithelial cell cultures was studied under 17-β-estradiol (100pg/ml) and progesterone (100ng/ml). u-PA mRNA increased in the presence of progesterone (P<0.05), but not by action of 17-β-estradiol. t-PA, PAI-1 and u-PAR were similar when cultured with the hormones. These results suggest that u-PA could be regulated by progesterone at a transcriptional level, by the balance of their activity for PAI-1 or at the epithelial surface through the binding of u-PAR. In conclusion, plasminogen activation system components might cooperate in the oviductal lumen to control plasmin generation.

New Unipolar Electrogram Criteria to Identify Irreversibility of Nonischemic Left Ventricular Cardiomyopathy

This study sought to assess the value of left ventricular (LV) endocardial unipolar electroanatomical mapping (EAM) in identifying irreversibility of LV systolic dysfunction in patients with left ventricular nonischemic cardiomyopathy (LVCM).

Redirecting Cell-type Specific Cytokine Responses with Engineered Interleukin-4 Superkines

Cytokines dimerize their receptors, with the binding of the 'second chain' triggering signaling. In the interleukin (IL)-4 and IL-13 system, different cell types express varying numbers of alternative second receptor chains (γc or IL-13Rα1), forming functionally distinct type I or type II complexes. We manipulated the affinity and specificity of second chain recruitment by human IL-4. A type I receptor-selective IL-4 'superkine' with 3,700-fold higher affinity for γc was three- to ten-fold more potent than wild-type IL-4. Conversely, a variant with high affinity for IL-13Rα1 more potently activated cells expressing the type II receptor and induced differentiation of dendritic cells from monocytes, implicating the type II receptor in this process. Superkines showed signaling advantages on cells with lower second chain numbers. Comparative transcriptional analysis reveals that the superkines induce largely redundant gene expression profiles. Variable second chain numbers can be exploited to redirect cytokines toward distinct cell subsets and elicit new actions, potentially improving the selectivity of cytokine therapy.

Mechanistic and Structural Insight into the Functional Dichotomy Between IL-2 and IL-15

Interleukin 15 (IL-15) and IL-2 have distinct immunological functions even though both signal through the receptor subunit IL-2Rβ and the common γ-chain (γ(c)). Here we found that in the structure of the IL-15-IL-15Rα-IL-2Rβ-γ(c) quaternary complex, IL-15 binds to IL-2Rβ and γ(c) in a heterodimer nearly indistinguishable from that of the IL-2-IL-2Rα-IL-2Rβ-γ(c) complex, despite their different receptor-binding chemistries. IL-15Rα substantially increased the affinity of IL-15 for IL-2Rβ, and this allostery was required for IL-15 trans signaling. Consistent with their identical IL-2Rβ-γ(c) dimer geometries, IL-2 and IL-15 showed similar signaling properties in lymphocytes, with any differences resulting from disparate receptor affinities. Thus, IL-15 and IL-2 induced similar signals, and the cytokine specificity of IL-2Rα versus IL-15Rα determined cellular responsiveness. Our results provide new insights for the development of specific immunotherapeutics based on IL-15 or IL-2.

HDAC Inhibition Decreases XIST Expression on Female IVP Bovine Blastocysts

During initial development, both X chromosomes are active in females, and one of them must be silenced at the appropriate time in order to dosage compensate their gene expression levels to male counterparts. Silencing involves epigenetic mechanisms, including histone deacetylation. Major X chromosome inactivation (XCI) in bovine occurs between hatching and implantation, although in vitro culture conditions might disrupt the silencing process, increasing or decreasing X-linked gene expression. In the present study, we aimed to address the roles of histone deacetylase inhibition by trichostatin A (TSA) on female preimplantation development. We tested the hypothesis that by enhancing histone acetylation, TSA would reduce percentage of embryos blocked at 8 cell stage, and interfere with XCI in IVF embryos. After TSA treatment, acetylation levels in individual blastomeres of 8-16 cell embryos were increased 2-fold on treated embryos, and the same was detected for blastocysts. Changes among blastomere levels were diminished on TSA group, as low-acetylated blastomeres were no longer detected. The percentage of embryos that reached the 5th cleavage cycle 118 h after IVF, analyzed by HOECHST staining, remained unaltered after TSA treatment. After, we assessed XIST and G6PD expression in individual female bovine blastocysts by real-time -qPCR. Even though G6PD remained unaltered after TSA exposure, XIST expression was 8-fold decreased, and we detected a major decrease in the percentage of blastocysts expressing detectable XIST levels after TSA treatment. Based on these results, we conclude that HDAC is involved on XCI process in bovine embryos, and its inhibition might delay X chromosome silencing and attenuate aberrant XIST expression described for IVF embryos.

Mechanical and Morphological Properties of Trabecular Bone Samples Obtained from Third Metacarpal Bones of Cadavers of Horses with a Bone Fragility Syndrome and Horses Unaffected by That Syndrome

Objective-To determine morphological and mechanical properties of trabecular bone of horses with a bone fragility syndrome (BFS; including silicate-associated osteoporosis). Sample-Cylindrical trabecular bone samples from the distal aspects of cadaveric third metacarpal bones of 39 horses (19 horses with a BFS [BFS bone samples] and 20 horses without a BFS [control bone samples]). Procedures-Bone samples were imaged via micro-CT for determination of bone volume fraction; apparent and mean mineralized bone densities; and trabecular number, thickness, and separation. Bone samples were compressed to failure for determination of apparent elastic modulus and stresses, strains, and strain energy densities for yield, ultimate, and failure loads. Effects of BFS and age of horses on variables were determined. Results-BFS bone samples had 25% lower bone volume fraction, 28% lower apparent density, 18% lower trabecular number and thickness, and 16% greater trabecular separation versus control bone samples. The BFS bone samples had 22% lower apparent modulus and 32% to 33% lower stresses, 10% to 18% lower strains, and 41 % to 52% lower strain energy densities at yield, ultimate, and failure loads, compared with control bone samples. Differences between groups of bone samples were not detected for mean mineral density and trabecular anisotropy. Conclusions and Clinical Relevance-Results suggested that horses with a BFS had osteopenia and compromised trabecular bone function, consistent with bone deformation and pathological fractures that develop in affected horses. Effects of this BFS may be systemic, and bones other than those that are clinically affected had changes in morphological and mechanical properties.

Relationship Between Body Fat and Masticatory Function

Purpose: The aim of this cross-sectional study was to investigate the relationship between body fat and masticatory function. Materials and Methods: One hundred dentate and partially edentulous participants (33 male; mean age, 39.7 ± 16.6 years) were selected. Body fat was established through body mass index (BMI). Masticatory function was evaluated by quantifying occlusal pairs and determining masticatory efficiency and swallowing threshold with the sieving method. During the swallowing threshold test, chewing rate was registered. Masticatory ability was also evaluated with a 5-point Likert scale questionnaire. Data were analyzed with Spearman and chi-square tests, as well as binary logistic regression analysis for the presence of increased BMI (α= 0.05). Results: Age (rho = 0.517), occlusal pairs (chi-square = 26.353), masticatory efficiency (chi-square = 30.935), masticatory ability (chi-square = 25.132; p < 0.001), and swallowing threshold (chi-square = 8.730; p < 0.005) were related to BMI. Age (odds ratio, OR = 1.048, 95% CI = 1.008 to 1.089) and lower masticatory efficiency (OR = 4.792, 95% CI = 1.419 to 16.183) were predictive of increased body fat (p < 0.05). Gender (chi-square = 0.402, p= 0.526) and chewing rate (rho =-0.158, p= 0.117) were not related to BMI. Conclusions: These results suggest that people with lower masticatory efficiency may be at risk for increased body fat.

Characterisation of Bound Volatile Compounds of a Low Flavour Kiwifruit Species: Actinidia Eriantha

Aroma compounds in fruit are known to occur in free and glycosidically bound forms. The bound volatile fraction of a low flavour kiwifruit species, Actinidia eriantha, was studied. The fruit have a bland and grassy flavour. Glycosidic precursors were isolated from juice by adsorption onto an Amberlite XAD-2 column. After enzymatic hydrolysis with Rapidase AR2000, the released aglycones were analysed by GC-MS. Alcohols, terpenoids and phenolics were the most numerously represented compound classes. Alcohols, benzenoids and phenolics showed the highest concentrations. Major compounds were 2-phenylethanol, furfuryl alcohol, (Z)-3-hexen-1-ol, coniferyl alcohol, isoamyl alcohol and linolenic acid. Several of the bound compounds found, including linoleic, linolenic and benzoic acids and coniferyl alcohol, are precursors of odorous volatiles. Many compounds detected as bound volatiles have not been previously reported as free volatiles in A. eriantha. The bound volatile composition of A. eriantha also showed differences with those of other kiwifruit species.

Protein Phosphorylation Regulates In vitro Spinach Chloroplast PetD MRNA 3'-untranslated Region Stability, Processing, and Degradation

RNA-binding proteins (RNPs) participate in diverse processes of mRNA metabolism, and phosphorylation changes their binding properties. In spinach chloroplasts, 24RNP and 28RNP are associated with polynucleotide posphorylase forming a complex on charge of pre-mRNA 3'-end maturation. Here, we tested the hypothesis that the phosphorylation status of 24RNP and 28RNP, present in a spinach chloroplast mRNA 3'-UTR processing extract (CPE), controls the transition between petD precursor stabilization, 3'-UTR processing, and RNA degradation in vitro. The CPE processed or stabilized petD precursor depending on the ATP concentration present in an in vitro 3'-UTR processing (IVP) assay. These effects were also observed when ATP was pre-incubated and removed before the IVP assay. Moreover, a dephosphorylated (DP)-CPE degraded petD precursor and recovered 3'-UTR processing or stabilization activities in an ATP concentration dependent manner. To determine the role 24/28RNP plays in regulating these processes a 24/28RNP-depleted (Δ24/28)CPE was generated. The Δ24/28CPE degraded the petD precursor, but when it was reconstituted with recombinant non-phosphorylated (NP)-24RNP or NP-28RNP, the precursor was stabilized, whereas when Δ24/28CPE was reconstituted with phosphorylated (P)-24RNP or P-28RNP, it recovered 3'-UTR processing, indicating that 24RNP or 28RNP is needed to stabilize the precursor, have a redundant role, and their phosphorylation status regulates the transition between precursor stabilization and 3'-UTR processing. A DP-Δ24/28CPE reconstituted or not with NP-24/28RNP degraded petD precursor. Pre-incubation of DP-Δ24/28CPE with NP-24/28RNP plus 0.03 mM ATP recovered 3'-UTR processing activity, and its reconstitution with P-24/28RNP stabilized the precursor. However, pre-incubation of DP-Δ24/28CPE with 0.03 mM ATP, and further reconstitution with NP-24/28RNP or P-24/28RNP produced precursor stability instead of RNA degradation, and RNA processing instead of precursor stability, respectively. Moreover, in vitro phosphorylation of CPE showed that 24RNP, 28RNP, and other proteins may be phosphorylated. Altogether, these results reveal that phosphorylation of 24RNP, 28RNP, and other unidentified CPE proteins mediates the in vitro interplay between petD precursor stability, 3'-UTR processing, and degradation, and support the idea that protein phosphorylation plays an important role in regulating mRNA metabolism in chloroplast.

Incidence and Predictors of Ovarian Function Recovery (OFR) in Breast Cancer (BC) Patients with Chemotherapy-induced Amenorrhea (CIA) Who Switched from Tamoxifen to Exemestane

BackgroundAromatase inhibitors (AIs) may promote ovarian function recovery (OFR). True incidence, predictors and impact on the outcome of OFR are unknown.Patients and methodsWe carried out a prospective study to assess ovarian function in estrogen receptor (ER)-positive BC patients on tamoxifen who had at least 2 years of chemotherapy-induced amenorrhea (CIA) and postmenopausal E2 levels. Patients switched to exemestane and underwent a series of investigations including vaginal ultrasound, antimullerian hormone, follicle stimulating hormone (FSH), and E2. E2 measurements were made using a clinical assay (direct) and a highly sensitive (indirect) immunoassay for comparison.ResultsBoth E2 assays (indirect versus direct) showed a similar incidence of OFR 32% (95% CI 19.5-44.5) versus 30% (95% CI 17.7-42.3) and median time to OFR 5.4 months (95% CI 1.2-9.6) versus 6.0 months (95% CI 4.8-7.1).On multivariate analysis, the mean age at the start of exemestane treatment was the only marker associated with probability of OFR (OR: 0.44, 0.24-0.78; P = 0.006). According to a receiver operating characteristic (ROC) analysis, age <48 years predicted for OFR (sensitivity: 59%; 1-specificity: 17%; AUC: 0.796; P = 0.001). Patients with OFR had higher mean E2 levels (43.6 versus 5.76 pmol/l; P = 0.001) and a reduced disease-free survival [DFS; HR 9.3 (95% CI 3.3-48.0; P = 0.04)] than those without it.ConclusionEven with a clinical and biochemical profile compatible with menopause, switching from tamoxifen to an AI should be avoided in patients <48 with CIA.

"The 3/3 Strategy": A Successful Multifaceted Hospital Wide Hand Hygiene Intervention Based on WHO and Continuous Quality Improvement Methodology

Only multifaceted hospital wide interventions have been successful in achieving sustained improvements in hand hygiene (HH) compliance.

Analysis of the Daily Changes of Melatonin Receptors in the Rat Liver

Melatonin membrane (MT1 and MT2) and nuclear (RORα) receptors have been identified in several mammalian tissues, including the liver. The mechanisms regulating hepatic melatonin receptors are yet unknown. This study investigated whether these receptors exhibit daily changes and the effects of melatonin on their levels. Our results show that mRNAs for MT1/MT2 receptors exhibit circadian rhythms that were followed by rhythms in their respective protein levels; the acrophases for the two rhythms were reached at 04:00 and 05:00 hr, respectively. Pinealectomy blunted the rhythms in both mRNAs and protein levels. In contrast, mRNA and protein levels of nuclear receptor RORα increased significantly after pinealectomy. The cycles of the latter receptor also exhibited circadian rhythms which peaked at 03:00 and 03:45 hr, respectively. Melatonin administration (10-200 mg/kg) increased in a dose-dependent manner the protein content of MT1/MT2 receptors, with no effects on RORα. Lunzindole treatment, however, did not affect melatonin receptor expression or content of either the membrane or nuclear receptors. Together with previously published findings which demonstrated the intracellular distribution of melatonin in rat liver, the current results support the conclusion that the circadian rhythms of MT1/MT2 and RORα receptors are under the control of the serum and intracellular melatonin levels. Moreover, the induction of MT1/MT2 receptors after the administration of high doses of melatonin further suggests that the therapeutic value of melatonin may not be restricted to only low doses of the indoleamine.

The Natural History of Compensated HCV-Related Cirrhosis; A Prospective Long-Term Study

BACKGROUND & AIMS: The natural history of HCV-related compensated cirrhosis has been poorly investigated in Latin-American countries. Our study evaluated mortality and clinical outcomes in compensated cirrhotic patients followed for 6 years. METHODS: Four hundred and two patients with compensated HCV-related cirrhosis were prospectively recruited in a tertiary care academic center. At the time of admission, patients were stratified as compensated (absence [stage 1], or presence [stage 2] of esophageal varices) as defined by D´Amico et al. Subjects were followed to identify overall mortality or liver transplantation and clinical complication rates. RESULTS: Among 402 subjects, 294 were categorized as stage 1 and 108 as stage 2. Over a median of 176 weeks, 42 deaths occurred (10%), of which 30 were considered liver-related (7%) and 12 non-liver-related (3%); 8 individuals (2%) underwent liver transplantation; 30 patients (7%) developed HCC, 67 individuals in stage 1 (22%) developed varices and any event of clinical decompensation occurred in 80 patients (20%). The 6-year cumulative overall mortality or liver transplantation was 15% and 45%, for stages 1 and 2, respectively (P<0.001). The cumulative 6-year HCC incidence was significantly higher among patients with varices (29%) than those without varices (9%), P<0.001. Similarly, the cumulative 6-year incidence of any clinical liver-related complication was higher in patients with stage 2 (66%) as compared to 26% in those with stage 1, respectively (P<0.001). CONCLUSIONS: Our results indicate significant morbidity and mortality and clinical outcome rates in compensated cirrhotic patients who have varices (stage 2).

Visible-Light Photocatalytic Hydrogen Generation by Using Dye-Sensitized Graphene Oxide As a Photocatalyst

Dye-sensitized graphene oxide is able to generate hydrogen from water/methanol mixtures (80:20) by using visible or solar light. The most efficient photocatalyst tested contained a tris(2,2-bipyridyl) ruthenium(II) complex incorporated in the interlayer spaces of a few layers of graphene oxide with a moderate degree of oxidation. The graphene oxide-based photocatalyst does not contain noble metals and we have determined that it is two orders of magnitude more active than catalysts based on conventional titania.

Gold Nanoparticles Downregulate Interleukin-1β-Induced Pro-Inflammatory Responses

Interleukin 1 beta (IL-1β)-dependent inflammatory disorders, such as rheumatoid arthritis and psoriasis, pose a serious medical burden worldwide, where patients face a lifetime of illness and treatment. Organogold compounds have been used since the 1930s to treat rheumatic and other IL-1β-dependent diseases and, though their mechanisms of action are still unclear, there is evidence that gold interferes with the transmission of inflammatory signalling. Here we show for the first time that citrate-stabilized gold nanoparticles, in a size dependent manner, specifically downregulate cellular responses induced by IL-1β both in vitro and in vivo. Our results indicate that the anti-inflammatory activity of gold nanoparticles is associated with an extracellular interaction with IL-1β, thus opening potentially novel options for further therapeutic applications.

Vision-based Sensor for Early Detection of Periodical Defects in Web Materials

During the production of web materials such as plastic, textiles or metal, where there are rolls involved in the production process, periodically generated defects may occur. If one of these rolls has some kind of flaw, it can generate a defect on the material surface each time it completes a full turn. This can cause the generation of a large number of surface defects, greatly degrading the product quality. For this reason, it is necessary to have a system that can detect these situations as soon as possible. This paper presents a vision-based sensor for the early detection of this kind of defects. It can be adapted to be used in the inspection of any web material, even when the input data are very noisy. To assess its performance, the sensor system was used to detect periodical defects in hot steel strips. A total of 36 strips produced in ArcelorMittal Avilés factory were used for this purpose, 18 to determine the optimal configuration of the proposed sensor using a full-factorial experimental design and the other 18 to verify the validity of the results. Next, they were compared with those provided by a commercial system used worldwide, showing a clear improvement.

Provision of Ubiquitous Tourist Information in Public Transport Networks

This paper outlines an information system for tourists using collective public transport based on mobile devices with limited computation and wireless connection capacities. In this system, the mobile device collaborates with the vehicle infrastructure in order to provide the user with multimedia (visual and audio) information about his/her trip. The information delivered, adapted to the user preferences, is synchronized with the passage of vehicles through points of interest along the route, for example: bus stops, tourist sights, public service centres, etc.

Ontological Representation of Light Wave Camera Data to Support Vision-Based AmI

Recent advances in technologies for capturing video data have opened a vast amount of new application areas in visual sensor networks. Among them, the incorporation of light wave cameras on Ambient Intelligence (AmI) environments provides more accurate tracking capabilities for activity recognition. Although the performance of tracking algorithms has quickly improved, symbolic models used to represent the resulting knowledge have not yet been adapted to smart environments. This lack of representation does not allow to take advantage of the semantic quality of the information provided by new sensors. This paper advocates for the introduction of a part-based representational level in cognitive-based systems in order to accurately represent the novel sensors' knowledge. The paper also reviews the theoretical and practical issues in part-whole relationships proposing a specific taxonomy for computer vision approaches. General part-based patterns for human body and transitive part-based representation and inference are incorporated to an ontology-based previous framework to enhance scene interpretation in the area of video-based AmI. The advantages and new features of the model are demonstrated in a Social Signal Processing (SSP) application for the elaboration of live market researches.

Uric Acid Stimulates Fructokinase and Accelerates Fructose Metabolism in the Development of Fatty Liver

Excessive dietary fructose intake may have an important role in the current epidemics of fatty liver, obesity and diabetes as its intake parallels the development of these syndromes and because it can induce features of metabolic syndrome. The effects of fructose to induce fatty liver, hypertriglyceridemia and insulin resistance, however, vary dramatically among individuals. The first step in fructose metabolism is mediated by fructokinase (KHK), which phosphorylates fructose to fructose-1-phosphate; intracellular uric acid is also generated as a consequence of the transient ATP depletion that occurs during this reaction. Here we show in human hepatocytes that uric acid up-regulates KHK expression thus leading to the amplification of the lipogenic effects of fructose. Inhibition of uric acid production markedly blocked fructose-induced triglyceride accumulation in hepatocytes in vitro and in vivo. The mechanism whereby uric acid stimulates KHK expression involves the activation of the transcription factor ChREBP, which, in turn, results in the transcriptional activation of KHK by binding to a specific sequence within its promoter. Since subjects sensitive to fructose often develop phenotypes associated with hyperuricemia, uric acid may be an underlying factor in sensitizing hepatocytes to fructose metabolism during the development of fatty liver.

Time-of-Flight-Secondary Ion Mass Spectrometry Method Development for High-Sensitivity Analysis of Acid Dyes in Nylon Fibers

A minimally destructive technique for the determination of dyes in finished fibers provides an important tool for crime scene and other forensic investigations. The analytical power and the minimal sample consumption of time-of-flight-secondary ion mass spectrometric (TOF-SIMS) analysis provides the ability to obtain definitive molecular and elemental information relevant to fiber identification, including identification of dyes, from a very small volume of sample. For both fiber surface analysis and, with the aid of cryomicrotomy, fiber cross-section analysis, TOF-SIMS was used to identify various dyes in finished textile fibers. The analysis of C.I. Acid Blue 25 in nylon is presented as a representative example. The molecular ion of C.I. Acid Blue 25 with lower than 3% on weight-of-fiber (owf) dye loading cannot be identified on dyed nylon surfaces by TOF-SIMS using a Bi(3)(+) primary ion beam. Sputtering with C(60)(+) provided the ability to remove surface contamination as well as at least partially remove Bi-induced damage, resulting in a greatly improved signal-to-noise ratio for the Acid Blue 25 molecular ion. The use of C(60)(+) for damage removal in a cyclic manner along with Bi for data acquisition provided the ability to unambiguously identify Acid Blue 25 via its molecular ion at a concentration of 0.1% owf from both fiber surfaces and cross sections.

Phylogeography of Mitochondrial Haplogroup D1: An Early Spread of Subhaplogroup D1j from Central Argentina

We analyzed the patterns of variation of haplogroup D1 in central Argentina, including new data and published information from other populations of South America. Almost 28% (107/388) of the individuals sampled in the region belong to haplogroup D1, whereas more than 52% of them correspond to the recently described subhaplogroup D1j (Bodner et al.: Genome Res 22 (2012) 811-820), defined by the presence of additional transitions at np T152C-C16242T-T16311C to the nodal D1 motif. This lineage was found at high frequencies across a wide territory with marked geographical-ecological differences. Additionally, 12 individuals present the mutation C16187T that defines the recently named subhaplogroup D1g (Bodner et al.: Genome Res 22 (2012) 811-820), previously described in populations of Patagonia and Tierra del Fuego. Based on our results and additional data already published, we postulate that the most likely origin of subhaplogroup D1j is the region of Sierras Pampeanas, which occupies the center and part of the northwestern portion of Argentina. The extensive yet restricted geographical distribution, the relatively large internal diversity, and the absence or low incidence of D1j in other regions of South America suggest the existence of an ancient metapopulation covering the Sierras Pampeanas, being this lineage its genetic signature. Further support for a scenario of local origin for D1j in the Sierras Pampeanas stems from the fact that early derivatives from a putative ancestral lineage carrying the transitions T16311C-T152C have only been found in this region, supporting the hypothesis that it might represent an ancestral motif previous to the appearance of D1j-specific change C16242T. © 2012 Wiley Periodicals, Inc.

Constitutively Active Mitogen-Activated Protein Kinase Versions Reveal Functions of Arabidopsis MPK4 in Pathogen Defense Signaling

Plant mitogen-activated protein kinases (MAPKs) are involved in important processes, including stress signaling and development. In a functional yeast screen, we identified mutations that render Arabidopsis thaliana MAPKs constitutively active (CA). Importantly, CA-MAPKs maintain their specificity toward known activators and substrates. As a proof-of-concept, Arabidopsis MAPK4 (MPK4) function in plant immunity was investigated. In agreement with the phenotype of mpk4 mutants, CA-MPK4 plants were compromised in pathogen-induced salicylic acid accumulation and disease resistance. MPK4 activity was found to negatively regulate pathogen-associated molecular pattern-induced reactive oxygen species production but had no impact on callose deposition, indicating that CA-MPK4 allows discriminating between processes regulated by MPK4 activity from processes indirectly affected by mpk4 mutation. Finally, MPK4 activity was also found to compromise effector-triggered immunity conditioned by the Toll Interleukin-1 Receptor-nucleotide binding (NB)-Leu-rich repeat (LRR) receptors RPS4 and RPP4 but not by the coiled coil-NB-LRR receptors RPM1 and RPS2. Overall, these data reveal important insights on how MPK4 regulates plant defenses and establishes that CA-MAPKs offer a powerful tool to analyze the function of plant MAPK pathways.

Inefficient Vaginal Transmission of Tenofovir Resistant HIV-1

Transmission of drug resistant HIV has been postulated to be a threat to current first line antiretroviral therapy (ART) regimes and the efficacy of several antiretroviral based pre-exposure prophylaxis (PrEP) strategies being tested. Here we evaluated the effect of the common tenofovir (TFV) resistance mutation, K65R on vaginal HIV transmission. Our results demonstrate that despite no overt loss of overall replication competence in vivo, this mutation results in significantly reduced mucosal transmission. When transmitted the mutant virus eventually reverted to wild type in 2 of 3 animals examined.

An Endohedral Metallofullerene As a Pure Electron Donor: Intramolecular Electron Transfer in Donor-Acceptor Conjugates of La(2)@C(80) and 11,11,12,12-Tetracyano-9,10-anthra-p-quinodimethane (TCAQ)

An endohedral metallofullerene, La(2)@C(80), is covalently linked to the strong electron acceptor 11,11,12,12-tetracyano-9,10-anthra-p-quinodimethane (TCAQ) by means of the Prato reaction, affording two different [5,6]-metallofulleropyrrolidines, namely 1a and 2a. 1a and 2a were isolated and fully characterized by means of MALDI-TOF mass, UV-vis-NIR absorption, and NMR spectroscopies. In addition, cyclic voltammetry (CV) and differential pulse voltammetry (DPV) corroborated the unique redox character of 2a, that is, the presence of the electron-donating La(2)@C(80) and the electron-accepting TCAQ. Although a weak electronic coupling dictates the interactions between La(2)@C(80) and TCAQ in the ground state, time-resolved transient absorption experiments reveal that in the excited state (i.e., π-π* centered at La(2)@C(80)) the unprecedented formation of the (La(2)@C(80))(•+)-(TCAQ)(•-) radical ion pair state evolves in nonpolar and polar media with a quantum efficiency of 33%.

Remodeling of the Folding Free Energy Landscape of Staphylococcal Nuclease by Cavity-creating Mutations

The folding of staphylococcal nuclease (SNase) is known to proceed via a major intermediate in which the central OB subdomain is folded and the C-terminal helical subdomain is disordered. To identify the structural and energetic determinants of this folding free energy landscape, we have examined in detail, using high-pressure NMR, the consequences of cavity creating mutations in each of the two subdomains of an ultrastable SNase, Δ+PHS. The stabilizing mutations of Δ+PHS enhanced the population of the major folding intermediate. Cavity creation in two different regions of the Δ+PHS reference protein, despite equivalent effects on global stability, had very distinct consequences on the complexity of the folding free energy landscape. The L125A substitution in the C-terminal helix of Δ+PHS slightly suppressed the major intermediate and promoted an additional excited state involving disorder in the N-terminus, but otherwise decreased landscape heterogeneity with respect to the Δ+PHS background protein. The I92A substitution, located in the hydrophobic OB-fold core, had a much more profound effect, resulting in a significant increase in the number of intermediate states and implicating the entire protein structure. Denaturant (GuHCl) had very subtle and specific effects on the landscape, suppressing some states and favoring others, depending upon the mutational context. These results demonstrate that disrupting interactions in a region of the protein with highly cooperative, unfrustrated folding has very profound effects on the roughness of the folding landscape, whereas the effects are less pronounced for an energetically equivalent substitution in an already frustrated region.

Tuberculin Skin Test Conversion Among Health Sciences Students: A Retrospective Cohort Study

Previous studies have reported that health sciences students are at greater risk for tuberculosis infection, especially in developing countries. The objective of this study was to estimate the prevalence, incidence, and factors associated with latent tuberculosis infection among Health Sciences students in Peru. Students enrolled at private university (in Lima - Peru) are tested annually for tuberculosis infection by tuberculin skin test. Data on tuberculin skin test results between 2002 and 2009 was used in this retrospective cohort study, a total of 4842 students were included. Tuberculin skin test conversion was defined as the change of tuberculin skin test from negative (<10 mm) to positive (≥10 mm) after 48-72 h of inoculation. Baseline tuberculin skin test positivity was 1.0% (95%CI: 0.6%-1.3%), whereas tuberculin skin test conversion incidence was 12.4 per 100 person-years (95%CI: 11.8-13.0). This study showed that students from clinical careers in close contact with patients had an increased risk of tuberculosis infection in the internship, especially Medicine, Dentistry, Medical Technology and Nursing. Administrative, environmental and personal protection measures should be implemented and evaluated periodically in order to reduce the risk of exposure.

Evidence for Chromosome 2p16.3 Polycystic Ovary Syndrome Susceptibility Locus in Affected Women of European Ancestry

Context:A previous genome-wide association study in Chinese women with polycystic ovary syndrome (PCOS) identified a region on chromosome 2p16.3 encoding the LH/choriogonadotropin receptor (LHCGR) and FSH receptor (FSHR) genes as a reproducible PCOS susceptibility locus.Objective:The objective of the study was to determine the role of the LHCGR and/or FSHR gene in the etiology of PCOS in women of European ancestry.Design:This was a genetic association study in a European ancestry cohort of women with PCOS.Setting:The study was conducted at an academic medical center.Participants:Participants in the study included 905 women with PCOS diagnosed by National Institutes of Health criteria and 956 control women.Intervention:We genotyped 94 haplotype-tagging single-nucleotide polymorphisms and two coding single-nucleotide polymorphisms mapping to the coding region of LHCGR and FSHR plus 20 kb upstream and downstream of the genes and test for association in the case control cohort and for association with nine quantitative traits in the women with PCOS.Results:We found strong evidence for an association of PCOS with rs7562215 (P = 0.0037) and rs10495960 (P = 0.0046). Although the marker with the strongest association in the Chinese PCOS genome-wide association study (rs13405728) was not informative in the European populations, we identified and genotyped three markers (rs35960650, rs2956355, and rs7562879) within 5 kb of rs13405728. Of these, rs7562879 was nominally associated with PCOS (P = 0.020). The strongest evidence for association mapping to FSHR was observed with rs1922476 (P = 0.0053). Furthermore, markers with the FSHR gene region were associated with FSH levels in women with PCOS.Conclusions:Fine mapping of the chromosome 2p16.3 Chinese PCOS susceptibility locus in a European ancestry cohort provides evidence for association with two independent loci and PCOS. The gene products LHCGR and FSHR therefore are likely to be important in the etiology of PCOS, regardless of ethnicity.

INFLUENCE OF EPISODES OF INTERMITTENT VIREMIA ("BLIPS") ON IMMUNE RESPONSES AND VIRAL LOAD REBOUND IN SUCCESSFULLY TREATED HIV-INFECTED PATIENTS

Background: Presenting episodes of intermittent viremia (EIV) under cART is frequent, but there exists some controversy about their consequences. They have been described to induce changes in immune responses potentially associated to a better control of HIV infection. Conversely, it has been suggested that EIV increase the risk of virological failure. Methods: A retrospective analysis of a prospective, randomized double blinded placebo-controlled study was performed. Twenty-six successfully treated HIV-infected adults were randomized to receive an immunization schedule or placebo, and after one year of follow-up cART was discontinued. Influence of EIV on T-cell subsets, HIV-1 specific T-cell immune responses, viral load rebound and the risk of developing genotypic mutations were evaluated, taking into account the immunization received. Results: Patients with EIV above 200 copies/mL under cART had a lower proportion of CD4+, CD4+CD45RA+RO- T-cells, a higher proportion of CD8+ and CD4+CD38+HLADR+ T-cells and higher HIV specific CD8+ T-cell responses compared to persistently undetectable patients. After cART interruption, patients with EIV presented a significantly higher viral rebound (p = 0.007), associated with higher increases in HIV specific lymphoproliferative responses and T-cell populations with activation markers. When patients with EIV between 20-200 copies/mL were included, most of the differences disappeared. Conclusion: Patients who present EIV above 200 copies/ml showed a lower CD4+ T-cell count and higher activation markers under cART. After treatment interruption, they showed higher specific immune responses against HIV that did not prevent a higher virologic rebound. EIV between 20 and 200 copies/ml did not have this deleterious effect.

Prognostic Role of MMP-9 Expression in Head and Neck Carcinoma Patients Treated with Radiotherapy or Chemoradiotherapy

OBJECTIVES: The purpose of this study was to evaluate the prognostic significance of the expression of metalloproteinases (MMPs)-2 and -9 at a transcriptional level in patients with head and neck squamous cell carcinoma (HNSCC). PATIENTS AND METHODS: Real-time polymerase chain reaction was used to determine mRNA expression levels of MMP-2 and MMP-9 in 105 consecutive patients with HNSCC treated with radiotherapy or chemoradiotherapy. Continuous values of mRNA expression levels were analyzed using a classification and regression tree (CART) method. RESULTS: Patients were grouped into two categories according to the results from CART analysis; high (n=71) and low (n=34) expression levels of MMP-9. MMP-2 expression was not included in the model. The 5-year adjusted survival rate was 92.9% for patients with low MMP-9 expression level and 61.0% for patients with a high expression level (P=0.006). Overexpression of MMP-9 was associated with a decrease in local control of the disease. In a multivariate analysis, MMP-9 expression was the only variable that was associated with adjusted survival. Considering patients with a low MMP-9 expression level as the reference group, patients with a high MMP-9 expression level had a 6.1 times higher risk of death from HNSCC (CI 95%: 1.4-26.4). CONCLUSION: We found a significant relationship between the transcription of MMP-9 and adjusted survival in HNSCC patients treated with radiotherapy or chemoradiotherapy. These results suggest that MMP-9 transcription may serve as a marker of treatment response to radiotherapy or chemo-radiotherapy in patients with HNSCC.

Renal Consequences of Intraabdominal Hypertension in a Porcine Model. Search for the Choice Indirect Technique for Intraabdominal Pressure Measurement

OBJECTIVE: To study the effects on the renal system in a porcine model of intraabdominal hypertension, and to determine the indirect technique of choice for determination of the intraabdominal pressure. MATERIAL AND METHODS: 30 pigs were used divided in two groups according with increased intraabdominal pressure values (20mmHg and 30mmHg). In both groups pressures were registered 8 times, summing up to 3hours, with a CO(2) insufflator. Three different measures of the intraabdominal pressure were taken: a direct transperitoneal measure, using a catheter of Jackson-Pratt connected to a pressure transducer, and two indirect measures, a transvesical by means of a Foley to manometer system, and a transgastric by introducing in the stomach a catheter connected to a pressure monitor with electronic hardware. Mean arterial pressure was calculated, along with the cardiac index, production of urine and serum creatinine. RESULTS: There was a greater correlation between the transvesical and the transperitoneal intraabdominal pressures (R(2)=0,95). Average transgastric intraabdominal pressure was inferior to the transperitoneal indicator in all taken measurements. The average arterial pressure descended in both groups, with earlier significant differences observed at 30mmHg (p<0,020). Urine production was lower at 30mmHg compared with the 20mmHg group (9,63±1,57 versus 3.26 ml±1,73). Serum creatinine increased in both groups being pathological at 30mmHg after 1h 20min, with existing differences between early pressures (p<0,027). CONCLUSIONS: This study revealed marked renal affectation with higher severity at 30mmHg pressures. The transvesical technique showed a greater correlation with the direct measurement technique used, defining this as the method of choice for determination of intraabdominal pressure.

Miniaturization of Hydrolase Assays in Thermocyclers

We adapted the protocols of reducing sugar measurements with dinitrosalicylic and bicinchoninic acid for thermocyclers, and their use in enzymatic assays for hydrolases such as amylase and β-1,3-glucanase. The use of thermocyclers for these enzymatic assays resulted in a 10 times reduction in the amount of reagent and volume of the sample needed, when compared to conventional microplate protocols. We standardized absorbance readings from the PCR plates which allowed us to make direct readings of the techniques above, and a β-glycosidase assay was also established under the same conditions. Standardization of the enzymatic reaction in thermocyclers resulted in less time-consuming temperature calibrations, and without loss of volume through leakage or evaporation from the microplate. Kinetic parameters were successfully obtained and the use of the thermocycler allowed the measurement of enzymatic activities in biological samples from the field with a limited amount of protein.

Smaller Quantal Size and Faster Kinetics of Single Exocytotic Events in Chromaffin Cells from the APP/PS1 Mouse Model of Alzheimer's Disease

The kinetics of single-amperometric exocytotic events has been measured in chromaffin cells of C57 mice and in an APP/PS1 mouse model of Alzheimer's disease (AD). K(+) depolarisation causes a burst of spikes that indicate the quantal release of the single-vesicle content of catecholamine. The kinetic analysis of 278 spikes from 10 control cells and 520 spikes from 18 APP/PS1 cells shows the following features of the latter compared with the former: (i) 45% lower t(1/2); (ii) 60% smaller quantal size; (iii) 50% lower decay time. Spike feet also showed 60% smaller quantal size. Immunofluorescence and thioflavin staining showed no amyloid beta (Aβ) burden in adrenal medulla slices of APP/PS1 mice that however exhibited dense Aβ plaques in the cortex and hippocampus. Furthermore, acetylcholinesterase staining of adrenal medulla indicated no apparent differences in the innervation by splanchnic cholinergic nerve terminals of chromaffin cells from control and APP/PS1 mice. This is the first report identifying subtle differences in the last steps of exocytosis that could be an indication of synaptic dysfunction of the secretory machinery not linked to Aβ burden in AD.

Micromotor-based Lab-on-chip Immunoassays

Here we describe the first example of using self-propelled antibody-functionalized synthetic catalytic microengines for capturing and transporting target proteins between the different reservoirs of a lab-on-a-chip (LOC) device. A new catalytic polymer/Ni/Pt microtube engine, containing carboxy moieties on its mixed poly(3,4-ethylenedioxythiophene) (PEDOT)/COOH-PEDOT polymeric outermost layer, is further functionalized with the antibody receptor to selectively recognize and capture the target protein. The new motor-based microchip immunoassay operations are carried out without any bulk fluid flow, replacing the common washing steps in antibody-based protein bioassays with the active transport of the captured protein throughout the different reservoirs, where each step of the immunoassay takes place. A first microchip format involving an 'on-the-fly' double-antibody sandwich assay (DASA) is used for demonstrating the selective capture of the target protein, in the presence of excess of non-target proteins. A secondary antibody tagged with a polymeric-sphere tracer allows the direct visualization of the binding events. In a second approach the immuno-nanomotor captures and transports the microsphere-tagged antigen through a microchannel network. An anti-protein-A modified microengine is finally used to demonstrate the selective capture, transport and convenient label-free optical detection of a Staphylococcus aureus target bacteria (containing proteinA in its cell wall) in the presence of a large excess of non-target (Saccharomyces cerevisiae) cells. The resulting nanomotor-based microchip immunoassay offers considerable potential for diverse applications in clinical diagnostics, environmental and security monitoring fields.

Evaluation of Neophobia and Its Potential Impact Upon Predator Control Techniques: A Study on Two Sympatric Foxes in Southern Patagonia

An alternative approach to increase the efficiency of predator control and selectivity is to consider the natural behavioural repertoire of the target species and how such behaviours may increase their vulnerability. Neophobia, or the hesitancy to approach a novel food item, object, or place, is an important factor influencing the investigative behaviour of animals, and its incorporation to predator control techniques may help to reduce losses of livestock to predators. In this study, we simultaneously evaluated the existence and intensity of neophobic responses in two sympatric fox species, the Culpeo (Pseudalopex culpaeus) and the Grey (P. griseus) foxes in southern Patagonia, Argentina. For this purpose, we used bait stations to compare fox behavioural responses in the absence (pre-treatment), presence (treatment) and removal (post-treatment) of a novel stimulus, which consisted of an orange PVC-traffic cone. Both fox species showed a neophobic response: bait-station visitation rates decreased (P=0.005 and P=0.048, for Culpeo and Grey foxes, respectively) in the presence of the novel object. The intensity of the response differed between species being higher for Culpeo foxes (approximately 80% of reduction in visitation rate during treatment for Culpeo foxes vs. 10% for Grey foxes). However, the bait-station visitation pattern after novel object removal indicated that animals probably increased exploration of the station. The high level of neophobia achieved by the Culpeo fox, together with an increase in post-treatment site exploration, suggests that behavioural manipulations (reduction of neophobia and its consequent increase in risk taking) could improve selective and efficient fox control in rural areas where livestock production is a major economic activity.

Synthesis, Antitumor, Antitrypanosomal and Antileishmanial Activities of Benzo[4,5]canthin-6-ones Bearing the N'-(Substituted Benzylidene)-carbohydrazide and N-Alkylcarboxamide Groups at C-2

A series of novel benzo[4,5]canthin-6-ones, bearing the N'-(substituted benzylidene)-carbohydrazide (11a-e) and N-alkylcarboxamide (13a-g) moieties at position-2, were synthesized and screened for their in vitro antitumor activity, against seven human cancer cell lines, and for antitrypanosomal and antileishmanial activities against Trypanosoma cruzi and Leishmania amazonensis. The results indicated that N-methylpiperazyl-6-oxobenzo[4,5]canthine-2-carboxamide (13f) displayed potent antitumor activity with IC(50) values in the range of 1.15-8.46 µM for all cell lines tested. Compounds 13f and 13g bearing an N-methylpiperazylcarboxamide and N-morpholylcarboxamide at C-2, respectively, showed potent activities towards both Trypanosoma cruzi and Leishmania amazonensis parasites, with IC(50) in the range of 0.4 to 16.70 µM.

Viral Encephalitis and Atherothrombotic Stroke

The clinical hallmark of viral encephalitis is an acute febrile illness, but stroke-like presentations have been rarely described. We report a patient who arrived at A&E with an acute right middle cerebral artery syndrome, without fever. Following thrombolytic treatment, clinical deficit almost disappeared and a critical ipsilateral carotid stenosis was identified. Stenting was implemented and 7 days later the patient started with high fever and reappearance of the initial deficit. Hyperperfusion syndrome and pneumonia were initially considered as the diagnosis but herpetic encephalitis (HE) was the final diagnosis. A very good response to antiviral treatment was achieved. We discuss whether the patient suffered from an unusual sudden stroke-like onset HE without encephalitic features or whether a stroke led to delayed HE.

Phosphorylation of Mitogen-activated Protein Kinases Contributes to IFN-γ Production in Response to Mycobacterium Tuberculosis

Immune control of M. tuberculosis depends on IFN-γ-producing CD4(+) lymphocytes, and previous studies have shown that T-cells from tuberculosis patients produce less IFN-γ in response to mycobacterial antigens than healthy donors, although IFN-γ responses to mitogens are preserved. In this work, we found that M. tuberculosis-induced IFN-γ production by human T-cells correlated with phosphorylation of the mitogen-activated protein kinases (MAPKs), extracellular signal-regulated kinase (ERK) and p38. Moreover, the majority of IFN-γ(+) T-cells expressed signaling lymphocyte activation molecule (SLAM), and SLAM activation further increased ERK phosphorylation. Interestingly, tuberculosis patients had delayed activation of ERK and p38, and this was most marked in patients with the poorest IFN-γ responses (low responders). Besides, SLAM signaling failed to phosphorylate ERK in low responder patients. Our findings suggest that activation of p38 and ERK, in part through SLAM, mediates T-cell IFN-γ production in response to M. tuberculosis, a pathway that is defective in tuberculosis patients.

Functional Evaluation Indicates Physical Losses After Hematopoietic Stem Cell Transplantation

To perform a function evaluation of patients before and after hematopoietic stem cell transplantation.

The Role of the Tolerance-fecundity Trade-off in Maintaining Intraspecific Seed Trait Variation in a Widespread Dimorphic Herb

Coexistence of species with different seed sizes is a long-standing issue in community ecology, and a trade-off between fecundity and stress tolerance has been proposed to explain co-occurrence in heterogeneous environments. Here we tested an intraspecific extension of this model: whether such trade-off also explains seed trait variation among populations of widespread plants under stress gradients. We collected seeds from 14 populations of Plantago coronopus along the Atlantic coast in North Africa and Europe. This herb presents seed dimorphism, producing large basal seeds with a mucilaginous coat that facilitates water absorption (more stress tolerant), and small apical seeds without coats (less stress tolerant). We analysed variation among populations in number, size and mucilage production of basal and apical seeds, and searched for relationships between local environment and plant size. Populations under higher stress (higher temperature, lower precipitation, lower soil organic matter) had fewer seeds per fruit, higher predominance of basal relative to apical seeds, and larger basal seeds with thicker mucilaginous coats. These results strongly suggest a trade-off between tolerance and fecundity at the fruit level underpins variation in seed traits among P. coronopus populations. However, seed production per plant showed the opposite pattern to seed production per fruit, and seemed related to plant size and other life-cycle components, as an additional strategy to cope with environmental variation across the range. The tolerance-fecundity model may constitute, under stress gradients, a broader ecological framework to explain trait variation than the classical seed size-number compromise, although several fecundity levels and traits should be considered to understand the diverse strategies of widespread plants to maximise fitness in each set of local conditions.

Efforts to Enhance Catheter Stability Improve Atrial Fibrillation Ablation Outcome

BACKGROUND: Contemporary techniques to enhance anatomical detail and catheter contact during atrial fibrillation ablation include: 1) the integration of pre-acquired tomographic reconstructions with electroanatomical mapping (I-EAM); 2) the use streerable introducers (SI); and 3) high frequency jet ventilation (HFJV). OBJECTIVE: We hypothesized that using these stabilizing techniques during AF ablation would improve 1-year procedural outcome. METHODS: We studied 300 patients undergoing AF ablation at our institution. Patients were assigned to three equal treatment groups (100 patients each) based upon the tools utilized: 1) Group 1- AF ablation performed without I-EAM, SI or HFJV; 2) Group 2- AF ablation performed using I-EAM and SI, but without HFJV; and 3) Group 3- AF ablation performed with I-EAM, SI, and HFJV. The primary outcome was freedom from AF 1-year after a single ablation procedure. The burden of both acute and chronic PV reconnection was also assessed. RESULTS: Patients from Groups 2 and 3 had significantly more non-paroxysmal AF (17% vs. 30% vs. 39%, p=0.002), larger left atria (4.2±0.8 vs. 4.4±0.7 vs. 4.5±0.8 cm, p<0.001), and higher BMI (28.5±5.8 vs. 29.1±4.8 vs. 31.2±5.4, p<0.001). Despite these differences, with adoption of I-EAM, SI, and HFJV we noted a significant improvement in 1-year freedom from AF (52% vs. 66% vs. 74%; p=0.006) as well as fewer acute (1.1±1.2 vs. 0.9±1.1 vs. 0.6±0.9, p=0.03) and chronic (3.5±0.9 vs. 3.2±0.9 vs. 2.4±1.0, p=0.02) PV reconnections. CONCLUSIONS: The incorporation of contemporary tools to enhance anatomical detail and ablation catheter stability significantly improved 1-year freedom from AF after ablation.

Cooperative Spin Transition in a Mononuclear Manganese(III) Complex

Mind the gap: A complete, cooperative spin transition for a mononuclear Mn(III) complex is reported with an 8 K hysteresis window. Raman spectra collected at a single temperature in warming and cooling modes confirm the electronic bistability within the hysteresis loop. The source of the cooperativity is a disconnection in the hydrogen-bonded 1D chains that connect adjacent cations owing to an order-disorder transition in the PF(6) (-) counterion.

A History of and a Vision for CMS Quality Measurement Programs

Tuning the Electronic Properties of Nonplanar ExTTF-Based Push-Pull Chromophores by Aryl Substitution

A new family of π-extended tetrathiafulvalene (exTTF) donor-acceptor chromophores has been synthesized by [2 + 2] cycloaddition of TCNE with exTTF-substituted alkynes and subsequent cycloreversion. X-ray data and theoretical calculations, performed at the B3LYP/6-31G** level, show that the new chromophores exhibit highly distorted nonplanar molecular structures with largely twisted 1,1,4,4-tetracyanobuta-1,3-diene (TCBD) units. The electronic and optical properties, investigated by UV/vis spectroscopy and electrochemical measurements, are significantly modified when the TCBD acceptor unit is substituted with a donor phenyl group, which increases the twisting of the TCBD units and reduces the conjugation between the two dicyanovinyl subunits. The introduction of phenyl substituents hampers the oxidation and reduction processes and, at the same time, largely increases the optical band gap. An effective electronic communication between the donor and acceptor units, although limited by the distorted molecular geometry, is evidenced both in the ground and in the excited electronic states. The electronic absorption spectra are characterized by low- to medium-intense charge-transfer bands that extend to the near-infrared.

Opportunistic Detection of Atrial Fibrillation in Subjects Aged 65 Years or Older in Primare Care: a Randomised Clinical Trial of Efficacy. DOFA-AP Study Protocol

ABSTRACT: BACKGROUND: Clinical Practice Guidelines recommend using peripheral blood pulse measuring as a screening test for Atrial Fibrillation. However, there is no adequate evidence supporting the efficacy of such procedure in primary care clinical practice. This paper describes a study protocol designed to verify whether early opportunistic screening for Atrial Fibrillation by measuring blood pulse is more effective than regular practice in subjects aged 65 years attending primary care centers. METHODS: An cluster-randomized controlled trial conducted in Primary Care Centers of the Spanish National Health Service. A total of 269 physicians and nurses will be allocated to one of the two arms of the trial by stratified randomization with a 3:2 ratio (three practitioners will be assigned to the Control Group for every two practitioners assigned to the Experimental Group). As many as 12 870 patients aged 65 years or older and meeting eligibility criteria will be recruited (8 580 will be allocated to the Experimental Group and 4 290 to the Control Group). Randomization and allocation to trial groups will be carried out by a central computer system. The Experimental Group practitioners will conduct an opportunistic case finding for patients with Atrial Fibrillation, while the Control Group practitioners will follow the regular guidelines. The first step will be finding new Atrial Fibrillation cases. A descriptive inferential analysis will be performed (bivariate and multivariate by multilevel logistic regression analysis). DISCUSSION: If our hypothesis is confirmed, we expect Primary Care professionals to take a more proactive approach and adopt a new protocol when a patient meeting the established screening criteria is identified. Finally, we expect this measure to be incorporated into Clinical Practice Guidelines.Trial registrationThe study is registered as NCT01291953 (ClinicalTrials.gob).

Low Co-existence Rates of Lactobacillus Spp. and Helicobacter Pylori Detected in Gastric Biopsies from Patients with Gastrointestinal Symptoms

Background: bacterial diversity of the stomach includes various species. Among them, Helicobacter pylori, a microorganism which has been associated to gastric diseases, is frequently isolated in this habitat. In addition, Lactobacillus spp., a genus including probiotic strains, has also been documented in this habitat. The co-existenceof these two species in the stomach of symptomatic patient needs to be elucidated.Aims: our goal was to establish if Lactobacillus spp. and H. pylori co-exist in the stomach mucosa of symptomatic patients.Methods: gastric biopsies (antrum and/or the body) from 427 Chilean patients with gastrointestinal discomfort were analyzed. The H. pylori infection and/or Lactobacillus spp. colonization status was determined for each patient by standard culture techniques, and statistical correlations between the presence of those species and the age, gender, or the severity of the gastric disease were also established.Results: only 6.1% of the samples presented co-existence of Lactobacillus spp. and H. pylori. This former species was isolated in 42.6% of the patients as unique species, while Lactobacillus spp. was isolated as single species in 19.4% of the individuals. Chronic non-atrophic gastritis was prevalent in Lactobacillus spp. non colonized individuals, while chronic non-atrophic and chronic atrophic gastritis diagnosis was similar in Lactobacillus spp. harbouring individuals (p < 0.001). The presence of Lactobacillus spp. Significantly increased with age (p = 0.005), independently of gender.Conclusion: the negative Pearson correlation between Lactobacillus spp. and H. pylori (r = -0.112, p = 0.020) indicates that the co-existence of both species is low in human gastric mucosa of symptomatic patients.

Adaptation of a Web-based, Open Source Electronic Medical Record System Platform to Support a Large Study of Tuberculosis Epidemiology

ABSTRACT: BACKGROUND: In 2006, we were funded by the US National Institutes of Health to implement a study of tuberculosis epidemiology in Peru. The study required a secure information system to manage data from a target goal of 16,000 subjects who needed to be followed for at least one year. With previous experience in the development and deployment of web-based medical record systems for TB treatment in Peru, we chose to use the OpenMRS open source electronic medical record system platform to develop the study information system. Supported by a core technical and management team and a large and growing worldwide community, OpenMRS is now being used in more than 40 developing countries.We adapted the OpenMRS platform to better support foreign languages. We added a new module to support double data entry, linkage to an existing laboratory information system, automatic upload of GPS data from handheld devices, and better security and auditing of data changes. We added new reports for study managers, and developed data extraction tools for research staff and statisticians. Further adaptation to handle direct entry of laboratory data occurred after the study was launched. RESULTS: Data collection in the OpenMRS system began in September 2009. By August 2011 a total of 9,256 participants had been enrolled, 102,274 forms and 13,829 laboratory results had been entered, and there were 208 users. The system is now entirely supported by the Peruvian study staff and programmers. CONCLUSIONS: The information system served the study objectives well despite requiring some significant adaptations mid-stream. OpenMRS has more tools and capabilities than it did in 2008, and requires less adaptations for future projects. OpenMRS can be an effective research data system in resource poor environments, especially for organizations using or considering it for clinical care as well as research.

Nucleofection of Whole Murine Retinas

The mouse retina constitutes an important research model for studies aiming to unravel the cellular and molecular mechanisms underlying ocular diseases. The accessibility of this tissue and its feasibility to directly obtain neurons from it has increased the number of studies culturing mouse retina, mainly retinal cell suspensions. However, to address many questions concerning retinal diseases and protein function, the organotypic structure must be maintained, so it becomes important to devise methods to transfect and culture whole retinas without disturbing their cellular structure. Moreover, the postmitotic stage of retinal neurons makes them reluctant to commonly used transfection techniques. For this purpose some published methods employ in vivo virus-based transfection techniques or biolistics, methods that present some constraints. Here we report for the first time a method to transfect P15-P20 whole murine retinas via nucleofection, where nucleic acids are directly delivered to the cell nuclei, allowing in vitro transfection of postmitotic cells. A detailed protocol for successful retina extraction, organotypic culture, nucleofection, histological procedures and imaging is described. In our hands the A-33 nucleofector program shows the highest transfection efficiency. Whole flat-mount retinas and cryosections from transfected retinas were imaged by epifluorescence and confocal microscopy, showing that not only cells located in the outermost retinal layers, but also those in inner retinal layers are transfected. In conclusion, we present a novel method to successfully transfect postnatal whole murine retina via nucleofection, showing that retina can be successfully nucleofected after some optimization steps.

Intergenerational Photovoice Projects: Optimizing This Mechanism for Influencing Health Promotion Policies and Strengthening Relationships

Intergenerational photovoice groups are promising for promoting health through the topic that is explored and through group dynamics that can foster healthy relationships and communication. To investigate the potential benefits of intergenerational photovoice projects, photovoice groups were conducted in urban Minnesota, United States, and in rural Morelos, Mexico, between 2009 and 2012 with Mexican-origin adults and their adolescent relatives. Seven photovoice groups of adult-adolescent dyads met for eight sessions and developed exhibits highlighting their views on health and migration and made policy recommendations, using messages conveyed through their words and photographs. Informal process evaluation and focus groups were used to elicit feedback about photovoice project participation. Photovoice project themes were descriptively analyzed. Focus group evaluation data were thematically summarized, and facilitator reflections were descriptively summarized to identify factors associated with intergenerational photovoice groups. Seventy-five participants were recruited. Photovoice themes represented effects of migration on health, family, and well-being. The following two evaluative themes were identified: (a) participant sentiments about the benefits of photovoice participation and (b) facilitator observations of intergenerational photovoice group benefits and challenges. Participants described opportunities to learn new things and barriers to healthy relationships that the project was eliminating by providing them with time to work together. Used in health promotion, photovoice is a valuable tool that contributes to understanding the complex underlying factors influencing behaviors and health.

Mortality Rates or Sociomedical Indicators? The Work of the League of Nations on Standardizing the Effects of the Great Depression on Health

This article explores the first international effort by the League of Nations Health Organization (LNHO) to standardize the study of the effects of the economic crisis of the 1930s on health. Instead of analysing this effort with the benefit of hindsight, this article takes into account the actors' perspectives and, therefore, it relies on the documents produced by the LNHO and public health experts of the 1930s, as well as on the historical scholarship on this subject. This article shows that, despite the declining death rates in Europe and in the US during the crisis, the LNHO considered that death rates concealed a more subtle effect of the crisis on health; hence, they launched a project aimed at making the effect visible. It describes the LNHO programme and the guidelines and methods set out by the organization in 1932 to observe this subtle effect through sociomedical investigations. The results of these surveys are summarized and the article discusses how the eugenic arguments used to explain them were not accepted by the LNHO. The article also shows how some members of the LNHO considered the results of the sociomedical surveys inconclusive and questioned the usefulness of socioeconomic indicators; in so doing, they raised concerns about the intervention of the LNHO in national matters and about the risks of crossing the established limits between science and politics. This article shows that an historical analysis, which takes into account the points of view of the actors involved, illuminates the factors that led the LNHO to conclude that mortality rates were the best method for measuring the effects of the economic crisis on health and that, as they were declining, the Great Depression was not having any deleterious effect on public health.

Metagenomic Analysis of the Microbiota from the Crop of an Invasive Snail Reveals a Rich Reservoir of Novel Genes

The shortage of petroleum reserves and the increase in CO(2) emissions have raised global concerns and highlighted the importance of adopting sustainable energy sources. Second-generation ethanol made from lignocellulosic materials is considered to be one of the most promising fuels for vehicles. The giant snail Achatina fulica is an agricultural pest whose biotechnological potential has been largely untested. Here, the composition of the microbial population within the crop of this invasive land snail, as well as key genes involved in various biochemical pathways, have been explored for the first time. In a high-throughput approach, 318 Mbp of 454-Titanium shotgun metagenomic sequencing data were obtained. The predominant bacterial phylum found was Proteobacteria, followed by Bacteroidetes and Firmicutes. Viruses, Fungi, and Archaea were present to lesser extents. The functional analysis reveals a variety of microbial genes that could assist the host in the degradation of recalcitrant lignocellulose, detoxification of xenobiotics, and synthesis of essential amino acids and vitamins, contributing to the adaptability and wide-ranging diet of this snail. More than 2,700 genes encoding glycoside hydrolase (GH) domains and carbohydrate-binding modules were detected. When we compared GH profiles, we found an abundance of sequences coding for oligosaccharide-degrading enzymes (36%), very similar to those from wallabies and giant pandas, as well as many novel cellulase and hemicellulase coding sequences, which points to this model as a remarkable potential source of enzymes for the biofuel industry. Furthermore, this work is a major step toward the understanding of the unique genetic profile of the land snail holobiont.

New Ideas in Lung Regeneration: a Personal View from Estoril

10th ERS Lung Science Conference - rebuilding a diseased lung: repair and regeneration Estoril, Portugal, 30 March-1 April 2012 The 10th ERS Lung Science Conference, held in Estoril, Portugal, focused on rebuilding a diseased lung: repair and regeneration, seeking to understand, with some amount of precision, how the processes by which the vastly complex self-assembling, self-repairing machine that is a human lung actually develops from a few cells in the embryo, repairs itself or fails to and succumbs to disease. Thus, the major research themes focused on lung development, lung stem and progenitor cell populations, regenerative signaling mechanisms and tissue engineering and transplantation.

Photochemical Evidence of Electronic Interwall Communication in Double-Wall Carbon Nanotubes

Single- and double-wall carbon nanotubes (CNTs) having dimethylanilino (DMA) units covalently attached to the external graphene wall have been prepared by the reaction of the dimethylaminophenylnitronium ion with the corresponding CNT. The samples have been characterized by Raman and XPS spectroscopies, thermogravimetry, and high-resolution transmission electron microscopy in which the integrity of the single or double wall of the CNT and the percentage of substitution (one dimethylanilino group every 45 carbons of the wall for the single- and double-wall samples) has been determined. Nanosecond laser flash photolysis has shown the generation of transients that has been derived from the charge transfer between the dimethylanilino (as the electron donor) to the CNT graphene wall (as the electron acceptor). Importantly, the lifetime of the double-wall CNT is much shorter than that monitored for the single-wall CNT. Shorter-lived transients were also observed for the pentyl-esterified functionalized double-wall CNT with respect to the single-wall analogue in the presence of hole (CH(3) OH) and electron quenchers (O(2) , N(2) O), which has led to the conclusion that the inner, intact graphene wall that is present in double-wall CNT increases the charge mobility significantly, favoring charge recombination processes. Considering the importance that charge mobility has in microelectronics, our finding suggests that double-wall CNT or two-layer graphene may be more appropriate to develop devices needing fast charge mobility.

Short-term Synaptic Plasticity Compensates for Variability in Number of Motor Neurons at a Neuromuscular Junction

We studied how similar postsynaptic responses are maintained in the face of interindividual variability in the number of presynaptic neurons. In the stomatogastric ganglion of the lobster, Homarus americanus, the pyloric (PY) neurons exist in variable numbers across animals. We show that each individual fiber of the stomach muscles innervated by PY neurons received synaptic input from all neurons present. We performed intracellular recordings of excitatory junction potentials (EJPs) in the muscle fibers to determine the consequences of differences in the number of motor neurons. Despite the variability in neuron number, the compound electrical response of muscle fibers to natural bursting input was similar across individuals. The similarity of total synaptic activation was not due to differences in the spiking activity of individual motor neurons across animals with different numbers of PY neurons. The amplitude of a unitary EJP in response to a single spike in a single motor neuron also did not depend on the number of PY neurons present. Consequently, the compound EJP in response to a single stimulus that activated all motor axons present was larger in individuals with more PY neurons. However, when axons were stimulated with trains of pulses mimicking bursting activity, EJPs facilitated more in individuals with fewer PY neurons. After a few stimuli, this resulted in depolarizations similar to the ones in individuals with more PY neurons. We interpret our findings as evidence that compensatory or homeostatic regulatory mechanisms can act on short-term synaptic dynamics instead of absolute synaptic strength.

Acute Mitral Stenosis After Transcatheter Aortic Valve Implantation

Inhibition of Taq DNA Polymerase by Iridoid Aglycone Derivates

Faithful replication of DNA molecules by DNA polymerases is essential for genome integrity and correct transmission of genetic information in all living organisms. DNA polymerases have recently emerged as important cellular targets for chemical intervention in the development of anti—cancer agents. Herein we report additional synthesis of simplified bicyclic aglycones of iridoids and their biological activity against Taq DNA polymerase with the object to find out some of the likely molecular targets implicated in the biological activity showed for this kind of compounds. The compounds 14, 33 and 34 showed inhibitory activity against Taq DNA polymerase with IC50 values of 13.47, 17.65 and 18.31 &mgr;M, respectively. These results would allow proposing to DNA polymerases as the molecular targets implicated in this bioactivity and enhance the iridoid aglycones as leader molecule to develop new drugs for cancer therapy.

Magnetic and Thermal Responses Triggered by Structural Changes in the Double Perovskite Sr(2)YRuO(6)

Among double perovskites, the interpretation of the magnetic, thermal and transport properties of Sr(2)YRuO(6) remains a challenge. Characterization using different techniques reveals a variety of features that are not understood, described as anomalous, and yields contradictory values for several relevant parameters. We solved this situation through detailed susceptibility, specific heat, thermal expansion and x-ray diffraction measurements, including a quantitative correlation of the parameters characterizing the so-called anomalies. The emergence of short-range magnetic correlations, surviving well above the long-range transition, naturally accounts for the observed unconventional behavior of this compound. High resolution x-ray powder diffraction and thermal expansion results conclusively show that the magnetic and thermal responses are driven by lattice changes, providing a comprehensive scenario in which the interplay between the spin and structural degrees of freedom plays a relevant role.

Florbetapir PET Analysis of Amyloid-β Deposition in the Presenilin 1 E280A Autosomal Dominant Alzheimer's Disease Kindred: a Cross-sectional Study

Fibrillar amyloid-β (Aβ) is thought to begin accumulating in the brain many years before the onset of clinical impairment in patients with Alzheimer's disease. By assessing the accumulation of Aβ in people at risk of genetic forms of Alzheimer's disease, we can identify how early preclinical changes start in individuals certain to develop dementia later in life. We sought to characterise the age-related accumulation of Aβ deposition in presenilin 1 (PSEN1) E280A mutation carriers across the spectrum of preclinical disease.

Compositional Uniformity, Domain Patterning and the Mechanism Underlying Nano-chessboard Arrays

We propose that systems exhibiting compositional patterning at the nanoscale, so far assumed to be due to some kind of ordered phase segregation, can be understood instead in terms of coherent, single phase ordering of minority motifs, caused by some constrained drive for uniformity. The essential features of this type of arrangement can be reproduced using a superspace construction typical of uniformity-driven orderings, which only requires the knowledge of the modulation vectors observed in the diffraction patterns. The idea is discussed in terms of a simple two-dimensional lattice-gas model that simulates a binary system in which the dilution of the minority component is favoured. This simple model already exhibits a hierarchy of arrangements similar to the experimentally observed nano-chessboard and nano-diamond patterns, which are described as occupational modulated structures with two independent modulation wavevectors and simple step-like occupation modulation functions.

Population Genetic Data of 30 Autosomal Indels in Central Spain and the Basque Country Populations

Samples from 71 unrelated Central Spain individuals and 60 Basque Country autochthonous individuals were typed with the Investigator DIPplex kit (30 biallelic autosomal mini-indels and amelogenin) and their allele frequencies were determined. Results demonstrated the assumption of independence within and between the loci analyzed. Different partially silent alleles were observed for the locus HLD97 (rs17238892) produced by a neighboring SNP (A/G), located 61bp downstream from the main indel site as shown by sequencing analysis.

Pre- and Postoperative Quantitative Analysis of Contour Abnormalities in Graves Upper Eyelid Retraction

PURPOSE:: One of the most common problems of the surgical management of Graves upper eyelid retraction is the occurrence of eyelid contour abnormalities. In the present study, the postoperative contour of a large sample of eyelids of patients with Graves orbitopathy was measured. METHODS:: The postoperative upper eyelid contour of 62 eyes of 43 patients with Graves orbitopathy was subjectively classified by 3 experienced surgeons in 3 categories: poor, fair, and good. The shape of the eyelid contours in each category was then measured with a recently developed custom-made software by measuring multiple midpupil eyelid distances each 15° along the palpebral fissure. The upper eyelid contour of 60 normal subjects was also quantified as a control group. RESULTS:: The mean ratio between the sum of the lateral and medial midpupil eyelid distances (lateral/medial ratio) was 1.10 ± 0.11 standard deviation in controls and 1.15 ± 0.13 standard deviation in patients. Postoperatively, the mean midpupil eyelid distance at 90° was 4.16 ± 1.13 mm standard deviation. The distribution lateral/medial ratios of the eyelids judged as having good contours was similar to the distribution of the controls with a modal value centered on the interval between 1.0 and 1.10. The distribution of lateral/medial ratios of the eyelids judged as having poor contour was bimodal, with eyelids with low and high lateral/medial ratios. Low lateral/medial ratios occurred when there was a lateral overcorrection, giving the eyelid a flat or a medial ptosis appearance. High lateral/medial ratios were due to a central or medial overcorrection or a lateral peak maintenance. CONCLUSIONS:: Postoperative upper eyelid contour abnormalities can be quantified by comparing the sum of multiple midpupil eyelid distances of the lateral and medial sectors of the eyelid. Low and high lateral/medial ratios are anomalous and judged as unpleasant.

In Vivo Accuracy of Conventional and Digital Radiographic Methods in Confirming Root Canal Working Length Determination by Root ZX

To compare, in vivo, the accuracy of conventional and digital radiographic methods in determining root canal working length.

Antioxidant and Cytotoxic Activities of Sideritis Perezlarae (Borja) Roselló, Stübing and Peris

Sideritis perezlarae is a plant widely used in folk medicine in the South of Andalusia (Cádiz, Spain). In this work, a phytochemical analysis has led to the isolation and identification of the flavonoid 7-O- β -glucosyl-luteolin from a methanol extract. The total phenol content estimated by Folin-Ciocalteau assay and expressed as gallic acid equivalent per gram of dried fraction, was 102.54 ± 2.10 mg phenols per gram dry residue. The flavonoid content, investigated by AlCl(3) reagent, was 23.49 ± 0.90 mg flavonoids gram dry residue. The methanol extract has been evaluated for antioxidant (DPPH and TEAC assays) and cytotoxic (SRB assay) properties. In the DPPH radical scavenging assay, the IC(50) was 360 µg mL(-1). In the total antioxidant activity, calculated by the Trolox equivalent antioxidant activity (TEAC, mg g(-1) of dried fraction), the extract showed a high antioxidant capacity (TEAC value of 0.59 ± 0.02 mg g(-1)). The cytotoxic activity of the extract against a human adenocarcinoma cell line HT-29 presented an IC(50) = 69.47 ± 4.64 µg mL(-1).

In Vivo Biodistribution of Amino-Functionalized Ceria Nanoparticles in Rats Using Positron Emission Tomography

A variety of nanoparticles have been proposed for several biomedical applications. To gauge the therapeutic potential of these nanoparticles, in vivo biodistribution is essential and mandatory. In the present study, ceria nanoparticles (5 nm average particle size) were labeled with (18)F to study their in vivo biodistribution in rats by positron emission tomography (PET). The (18)F isotope was anchored by reaction of N-succinimidyl 4-[(18)F]fluorobenzoate ((18)F-SFB) with a modified nanoparticle surface obtained by silylation with 3-aminopropylsilyl. Radiolabeled ceria nanoparticles accumulated mainly in lungs, spleen, and liver. Metabolic products of the radiolabeled nanoparticulate material were excreted into the urinary tract.

Minor Myocardial Damage is a Prevalent Condition in Patients With Acute Heart Failure Syndromes and Preserved Systolic Function With Long-Term Prognostic Implications. A Report From the CIAST-HF (Collaborative Italo-Argentinean Study on Cardiac Troponin T in Heart Failure) Study

Half of patients with acute heart failure syndromes (AHFS) have preserved left ventricular ejection fraction (PLVEF). In this setting, the role of minor myocardial damage (MMD), as identified by cardiac troponin T (cTnT), remains to be established.

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Insights into the Function of RifI2: Structural and Biochemical Investigation of a New Shikimate Dehydrogenase Family Protein

The shikimate dehydrogenase (SDH) family consists of enzymes with diverse roles in secondary metabolism. The two most widespread members of the family, AroE and YdiB, function in amino acid biosynthesis and quinate catabolism, respectively. Here, we have determined the crystal structure of an SDH homolog belonging to the RifI class, a group of enzymes with proposed roles in antibiotic biosynthesis. The structure of RifI2 from Pseudomonas putida exhibits a number of distinctive features, including a substantial C-terminal truncation and an atypical mode of oligomerization. The active site of the enzyme contains substrate- and cofactor-binding motifs that are significantly different from those of any previously characterized member of the SDH family. These features are reflected in the novel kinetic properties of the enzyme. RifI2 exhibits much lower activity using shikimate as a substrate than AroE, and a strong preference for NAD(+) instead of NADP(+) as a cofactor. Moreover, the enzyme has only trace activity using quinate, unlike YdiB. Cocrystallization of RifI2 with NAD(+) provided the opportunity to determine the mode of cofactor selectivity employed by the enzyme. We complemented this analysis by probing the role of a strictly conserved residue in the cofactor-binding domain, Asn193, by site directed mutagenesis. This study presents the first crystal structure and formal kinetic characterization of a new NAD(+)-dependent member of the SDH family.

Radiological Evaluation of Acetabular Erosion After Antibiotic-Impregnated Polymethylmethacrylate Spacer (Spacer-G)

Different types of hip spacers have been described (hand-made, custom-molded or prefabricated) for treatment of a chronic hip infection. A potential disadvantage of monoblock prefabricated spacer is that it may cause acetabular bone loss. This study assesses the radiological acetabular erosion using an antibiotic-impregnated pre-fabricated polymethylmethacrylate Spacer-G. We retrospectively reviewed the radiographs of thirty five patients who were managed with Spacer-G to treat chronic hip infection. No acetabular erosion were observed in thirty two patients with a mean time from the first to second stage and from the first to the last radiograph of 5.09 and 3.77months respectively. In three patients the time between the radiographs was more than one year and the second stage was not performed; two developed a protrusion acetabuli whereas the other one a destruction of the acetabular roof. Using a Spacer-G in chronic hip infection treatment for less than one year is not associated with radiological acetabular erosion if the patient is maintained at partial weight bearing.

[Initial Situation and Approach to the Care of Migraine in Neurology Services in Spain. The PRIMERA Study]

INTRODUCTION. Despite the high degree of disability it entails, many patients with migraine have never visited their doctor for this reason. It is necessary to conduct a study to examine the characteristics of first-time visits as a step that must be carried out prior to establishing specific intervention measures for this group of patients. AIM. To determine the profile of the patients with migraine who visit a neurology service for the first time, together with the diagnostic and therapeutic attitudes that neurologists display towards them. PATIENTS AND METHODS. We conducted a cross-sectional, multi-centre study of neurology services across the country. The research included 168 neurologists who recruited 851 patients (74.6% females; mean age: 34.0 ± 10.7 years). Disability was assessed by means of the specific migraine questionnaire (Headache Impact Test) and the generic disability questionnaire (Sheehan Disability Scale). RESULTS. A third (66.5%) of the patients went for consultation following their doctor's advice, while the remaining 33.5% went on their own accord. Only 55.9% had been previously diagnosed with migraine. The main reasons for visiting were ineffective symptomatic treatment (25%) and an increase in the frequency or intensity of the attacks (23.4%). Although 70.3% of the patients had high disability scores on the Headache Impact Test, only 17.4% used specific treatment and only 13.3% were on preventive treatment. CONCLUSIONS. The PRIMERA study confirms, once again, that migraine is an under-diagnosed and under-treated condition in our setting, which means that specific educational interventions and training are still required for this pathology.

Occupational Allergic Rhinoconjunctivitis and Bronchial Asthma Induced by Goat Cheese

Cysteine-Generated Sulfide in the Cytosol Negatively Regulates Autophagy and Modulates the Transcriptional Profile in Arabidopsis

In Arabidopsis thaliana, DES1 is the only identified l-Cysteine desulfhydrase located in the cytosol, and it is involved in the degradation of cysteine (Cys) and the concomitant production of H(2)S in this cell compartment. Detailed characterization of the T-DNA insertion mutants des1-1 and des1-2 has provided insight into the role of sulfide metabolically generated in the cytosol as a signaling molecule. Mutations of L-CYS DESULFHYDRASE 1 (DES1) impede H(2)S generation in the Arabidopsis cytosol and strongly affect plant metabolism. Senescence-associated vacuoles are detected in mesophyll protoplasts of des1 mutants. Additionally, DES1 deficiency promotes the accumulation and lipidation of the ATG8 protein, which is associated with the process of autophagy. The transcriptional profile of the des1-1 mutant corresponds to its premature senescence and autophagy-induction phenotypes, and restoring H(2)S generation has been shown to eliminate the phenotypic defects of des1 mutants. Moreover, sulfide is able to reverse ATG8 accumulation and lipidation, even in wild-type plants when autophagy is induced by carbon starvation, suggesting a general effect of sulfide on autophagy regulation that is unrelated to sulfur or nitrogen limitation stress. Our results suggest that Cys-generated sulfide in the cytosol negatively regulates autophagy and modulates the transcriptional profile of Arabidopsis.

DNA Hypomethylation Affects Cancer-related Biological Functions and Genes Relevant in Neuroblastoma Pathogenesis

Neuroblastoma (NB) pathogenesis has been reported to be closely associated with numerous genetic alterations. However, underlying DNA methylation patterns have not been extensively studied in this developmental malignancy. Here, we generated microarray-based DNA methylation profiles of primary neuroblastic tumors. Stringent supervised differential methylation analyses allowed us to identify epigenetic changes characteristic for NB tumors as well as for clinical and biological subtypes of NB. We observed that gene-specific loss of DNA methylation is more prevalent than promoter hypermethylation. Remarkably, such hypomethylation affected cancer-related biological functions and genes relevant to NB pathogenesis such as CCND1, SPRR3, BTC, EGF and FGF6. In particular, differential methylation in CCND1 affected mostly an evolutionary conserved functionally relevant 3' untranslated region, suggesting that hypomethylation outside promoter regions may play a role in NB pathogenesis. Hypermethylation targeted genes involved in cell development and proliferation such as RASSF1A, POU2F2 or HOXD3, among others. The results derived from this study provide new candidate epigenetic biomarkers associated with NB as well as insights into the molecular pathogenesis of this tumor, which involves a marked gene-specific hypomethylation.

Generation and Characterization of a Defective HIV-1 Virus As an Immunogen for a Therapeutic Vaccine

The generation of new immunogens able to elicit strong specific immune responses remains a major challenge in the attempts to obtain a prophylactic or therapeutic vaccine against HIV/AIDS. We designed and constructed a defective recombinant virus based on the HIV-1 genome generating infective but non-replicative virions able to elicit broad and strong cellular immune responses in HIV-1 seropositive individuals.

Transplant Glomerulopathy: Clinical Course and Factors Relating to Graft Survival

Transplant glomerulopathy (TG) is usually associated with a poor prognosis for kidney graft survival.

Accuracy of Noninvasive Estimation of Pulmonary Wedge Pressure by Echocardiographic Indices in Heart Transplant Recipients

Pulmonary capillary wedge pressure (PCWP) can be estimated from transmitral or color M-mode Doppler flow propagation velocities. However, it has been recommended to not use these indices in heart transplant recipients. Our aim was to compare the accuracy of color M-mode, Doppler, and Doppler tissue imaging (DTI)-derived indices to predict PCWP in heart transplant recipients.

Antipsychotics and Seizures: Higher Risk with Atypicals?

PURPOSE: Almost all antipsychotics have been associated with a risk of epileptic seizure provocation. Among the first-generation antipsychotics (FGA) chlorpromazine appears to be associated with the greatest risk of seizures among the second-generation antipsychotics (SGA) clozapine is thought to be most likely to cause convulsions. This information is largely based on studies that are not sufficiently controlled. Besides, information about the seizure risk associated with newer antipsychotics is scarce. METHOD: The Pharmacovigilance Unit of the Basque Country (network of centers of the Spanish Pharmacovigilance System, SEFV) provided reporting data for adverse reactions (AR) from the whole SEFV to estimate the reporting odds ratio (ROR) for antipsychotics and seizures ("convulsions" as Single MedDra Query). Data was obtained from SEFV database from 1984 to the June 2011. RESULTS: The total number of convulsions reported for SGA was 169 (total reported AR 3.204). The number of convulsions reported for FGA was 35 (total reported AR 2.051). 94 convulsions were reported in association with clozapine (total AR 1.052). The ROR for SGA versus FGA was 3.2 (CI 95%: 2.21-4.63). The ROR for SGA excluding clozapine versus FGA was 2.08 (CI 95%: 1.39-3.12). CONCLUSION: Our results show that SGA may pose a higher risk of seizures than FGA, mainly, but not only due to clozapine. This is line with recent studies suggesting that some SGA carried a higher average risk of electroencephalographic abnormalities than many FGA. Nonetheless, It is well known that spontaneous reports do not allow strong inferences about adverse drug effects, because differences in reporting fractions by time, drug or type of event are difficult or even impossible to distinguish from differences in the occurrence rates of adverse events. Still, we consider that the possibility of SGA carrying a higher risk of seizure induction than FGA warrants further research.

Comparative Cost Models of a Liquid Nitrogen Vapor Phase (LNVP) Cold Chain-distributed Cryopreserved Malaria Vaccine Vs. a Conventional Vaccine

Typically, vaccines distributed through the Expanded Program on Immunization (EPI) use a 2-8°C cold chain with 4-5 stops. The PfSPZ vaccine comprises whole live-attenuated cryopreserved sporozoites stored in liquid nitrogen (LN(2)) vapor phase (LNVP) below -140°C and would be distributed through a LNVP cold chain. The purpose of this study was to model LNVP cold chain distribution for the cryopreserved PfSPZ Vaccine in Tanzania, estimate the costs and compare these costs to those that would be incurred in distributing a 'conventional' malaria vaccine through the EPI. Capital and recurrent costs for storage, transportation, labor, energy usage and facilities were determined for the birth cohort in Tanzania over five years. Costs were calculated using WHO/UNESCO calculators. These were applied to a 2-8°C distribution model with national, regional, district, and health facility levels, and for the cryopreserved vaccine using a 'modified hub-and-spoke' (MH-S) LNVP distribution system comprising a central national store, peripheral health facilities and an intermediate district-level transhipment stop. Estimated costs per fully immunized child (FIC) were $ 6.11 for the LNVP-distributed cryopreserved vaccine where the LN(2) is generated, and $ 6.04 with purchased LN(2) (assuming US $ 1.00/L). The FIC costs for distributing a conventional vaccine using the four level 2-8°C cold chain were $ 6.10, and with a tariff distribution system as occurs in Tanzania the FIC cost was $ 5.53. The models, therefore, predicted little difference in 5-year distribution costs between the PfSPZ vaccine distributed through a MH-S LNVP cold chain and a conventional vaccine distributed through the more traditional EPI system. A LNVP cold chain provides additional benefits through the use of durable dry shippers because no refrigerators, freezers or refrigerated trucks are required. Thus strain at the cold chain periphery, vaccine wastage from cold chain failures and the environmental impact of distribution would all be reduced.

The Chest and Aging: Radiological Findings

In the elderly (conventionally defined as individuals > 60 years of age), it is often difficult to establish what normality is, because of the numerous anatomical and physiological modifications that occur during the aging process. As a result, the greatest challenge is to differentiate between the normal aging process and the onset of disease. Healthy elderly people commonly present borderline findings on chest imaging. We systematically reviewed the medical literature on the subject, covering the period between 1950 and 2011, including articles in Portuguese, English, French, Italian, and Spanish. We searched the PubMed, LILACS, and SciELO databases, using the search terms "age", "aging", "lung", "thorax", "chest", "X-ray", "radiography", "pulmonary", and "computed tomography"-as well as their corresponding translations-in various combinations. We included only original or review articles on aging-related chest imaging findings. In broad terms, aging results in physiological modifications that must be recognized so as not to be erroneously interpreted as pathological.

Efficient Serum-resistant Lipopolyplexes Targeted to the Folate Receptor

In this work, we have developed and evaluated a new targeted lipopolyplex (LPP), by combining polyethylenimine (PEI), 1,2-dioleoyl-3-(trimethylammonium) propane (DOTAP)/Chol liposomes, the plasmids pCMVLuc/pCMVIL-12, and the ligand folic acid (FA), able to transfect HeLa and B16-F10 cells in the presence of very high concentration of serum (60% FBS). These complexes (Fol-LPP) have a net positive surface charge. The combination of folic acid with lipopolyplexes also enhanced significantly the transfection activity of the therapeutic gene interleukin-12 (IL-12), without any significant cytotoxicity. The specificity of the folate receptor (FR)-mediated gene transfer was corroborated by employing a folate receptor deficient cell line (HepG2). This formulation improved gene delivery showed by conventional lipoplexes or polyplexes resulting an efficient, simple, and nontoxic method for gene delivery of therapeutic genes in vitro and very probably in vivo.

[Validation Inspection of Prescription Drugs in Spain: Current Situation from a Health Professional Perspective After Its Introduction in Electronic Form.]

AIMS: To assess the current situation of the inspection validation of prescriptions (IVP) in Spain since the introduction of the electronic procedure (EP) from the healthcare professionals' perspective. DESIGN: Observational, cross sectional study, in two phases; combining qualitative and quantitative techniques. SETTING: Primary Care (PC), Secondary Care (SC) and the health care management sector. PARTICIPANTS: Primary care physicians (PCPs), endocrinologists, medical inspectors, pharmacists and health Authorities of Autonomous Communities (AACC). METHOD: Semi-structured surveys and Computer Assisted Telephone Interviewing. RESULTS: The IVP presents three modalities in Spain: manual, electronics and electronics linked to electronic prescription. The participants of the same level of management perceive in a similar way the purpose of the IVP, and there exist differences between the different levels of interviewed managers. Differences exist in the situation of EP between AACC. It is more implemented in primary care (PC) than in specialized (63% vs 37%), with similar degree in urban and rural areas. Six of 17 AACC presented a public access to the corresponding legislation. CONCLUSION: The IVP is perceived as a tool for the economic control in expenditure on drugs and as additional administrative load in PC. The rhythm of implementation of EP differs between AACC as well as the access to the regulation.

Comparison of PET Metabolic Indices for the Early Assessment of Tumour Response in Metastatic Colorectal Cancer Patients Treated by Polychemotherapy

PURPOSE: To compare the performance of eight metabolic indices for the early assessment of tumour response in patients with metastatic colorectal cancer (mCRC) treated with chemotherapy. METHODS: Forty patients with advanced mCRC underwent two FDG PET/CT scans, at baseline and on day 14 after chemotherapy initiation. For each lesion, eight metabolic indices were calculated: four standardized uptake values (SUV) without correction for the partial volume effect (PVE), two SUV with correction for PVE, a metabolic volume (MV) and a total lesion glycolysis (TLG). The relative change in each index between the two scans was calculated for each lesion. Lesions were also classified as responding and nonresponding lesions using the Response Evaluation Criteria In Solid Tumours (RECIST) 1.0 measured by contrast-enhanced CT at baseline and 6-8 weeks after starting therapy. Bland-Altman analyses were performed to compare the various indices. Based on the RECIST classification, ROC analyses were used to determine how accurately the indices predicted lesion response to therapy later seen with RECIST. RESULTS: RECIST showed 27 responding and 74 nonresponding lesions. Bland-Altman analyses showed that the four SUV indices uncorrected for PVE could not be used interchangeably, nor could the two SUV corrected for PVE. The areas under the ROC curves (AUC) were not significantly different between the SUV indices not corrected for PVE. The mean SUV change in a lesion better predicted lesion response without than with PVE correction. The AUC was significantly higher for SUV uncorrected for PVE than for the MV, but change in MV provided some information regarding the lesion response to therapy (AUC >0.5). CONCLUSION: In these mCRC patients, all SUV uncorrected for PVE accurately predicted the tumour response on day 14 after starting therapy as assessed 4 to 6 weeks later (i.e. 6 to 8 weeks after therapy initiation) using the RECIST criteria. Neither correcting SUV for PVE nor measuring TLG improved the assessment of tumour response compared to SUV uncorrected for PVE. The change in MV was the least accurate index for predicting tumour response.

In Vitro Effects of the CCR5 Inhibitor Maraviroc on Human T Cell Function

BACKGROUND: Several potential immunological benefits have been observed during treatment with the CC chemokine receptor 5 (CCR5) antagonist maraviroc, in addition to its antiviral effect. Our objective was to analyse the in vitro effects of CCR5 blockade on T lymphocyte function and homeostasis. METHODS: Peripheral blood mononuclear cells (PBMCs) from both HIV-negative (n = 28) and treated HIV-positive (n = 27) individuals were exposed in vitro to different concentrations of maraviroc (0.1-100 μM). Effects on T cell activation were analysed by measuring the expression of the CD69, CD38, HLA-DR and CD25 receptors as well as CCR5 density using flow cytometry. Spontaneous and chemokine-induced chemotaxis were measured by transwell migration assays, and polyclonal-induced proliferation was assessed by a lymphoproliferation assay and carboxyfluorescein succinimidyl ester staining. RESULTS: Maraviroc increases CCR5 surface expression on activated T cells, even at low doses (0.1 μM). Slight differences were detected in the frequency and mean fluorescence intensity of activation markers at high concentrations of maraviroc. Expression of CD25, CD38 and HLA-DR tended to decrease in both CD4+ and CD8+ T lymphocytes, whereas expression of CD69 tended to increase. Maraviroc clearly inhibits T cell migration induced by chemokines in a dose-dependent manner. Moreover, at 100 μM, maraviroc tends to inhibit T cell proliferation. CONCLUSIONS: These data showed that in vitro exposure to maraviroc decreases some activation expression markers on T lymphocytes and also migration towards chemoattractants. These results support the additional immunological effects of CCR5 blockade and suggest that maraviroc might have potential capacity to inhibit HIV-associated chronic inflammation and activation, both by directly affecting T cell activation and by reducing entrapment of lymphocytes in lymph nodes.

Molecular Characterization of Chronic Lymphocytic Leukemia Patients with a High Number of Losses in 13q14

Patients with chronic lymphocytic leukemia and 13q deletion as their only FISH abnormality could have a different outcome depending on the number of cells displaying this aberration. Thus, cases with a high number of 13q- cells (13q-H) had both shorter overall survival and time to first therapy. The goal of the study was to analyze the genetic profile of 13q-H patients.

Counteracting Roles of AMP Deaminase and AMP Kinase in the Development of Fatty Liver

Fatty liver (hepatic steatosis) is associated with nucleotide turnover, loss of ATP and generation of adenosine monophosphate (AMP). It is well known that in fatty liver, activity of the AMP-activated kinase (AMPK) is reduced and that its stimulation can prevent hepatic steatosis by both enhancing fat oxidation and reducing lipogenesis. Here we show that another AMP dependent enzyme, AMPD2, has opposing effects on fatty acid oxidation when compared to AMPK. In human hepatocytres, AMPD2 activation -either by overexpression or by lowering intracellular phosphate levels with fructose- is associated with a significant reduction in AMPK activity. Likewise, silencing of AMPK spontaneously increases AMPD activity, demonstrating that these enzymes counter-regulate each other. Furthermore, we show that a downstream product of AMP metabolism through AMPD2, uric acid, can inhibit AMPK activity in human hepatocytes. Finally, we show that fructose-induced fat accumulation in hepatocytes is due to a dominant stimulation of AMPD2 despite stimulating AMPK. In this regard, AMPD2-deficient hepatocytes demonstrate a further activation of AMPK after fructose exposure in association with increased fatty acid oxidation, and conversely silencing AMPK enhances AMPD-dependent fat accumulation. In vivo, we show that sucrose fed rats also develop fatty liver that is blocked by metformin in association with both a reduction in AMPD activity and an increase in AMPK activity. In summary, AMPD and AMPK are both important in hepatic fat accumulation and counter-regulate each other. We present the novel finding that uric acid inhibits AMPK kinase activity in fructose-fed hepatocytes thus providing new insights into the pathogenesis of fatty liver.

Compartmentalization of Mammalian Pantothenate Kinases

The pantothenate kinases (PanK) catalyze the first and the rate-limiting step in coenzyme A (CoA) biosynthesis and regulate the amount of CoA in tissues by differential isoform expression and allosteric interaction with metabolic ligands. The four human and mouse PanK proteins share a homologous carboxy-terminal catalytic domain, but differ in their amino-termini. These unique termini direct the isoforms to different subcellular compartments. PanK1α isoforms were exclusively nuclear, with preferential association with the granular component of the nucleolus during interphase. PanK1α also associated with the perichromosomal region in condensing chromosomes during mitosis. The PanK1β and PanK3 isoforms were cytosolic, with a portion of PanK1β associated with clathrin-associated vesicles and recycling endosomes. Human PanK2, known to associate with mitochondria, was specifically localized to the intermembrane space. Human PanK2 was also detected in the nucleus, and functional nuclear localization and export signals were identified and experimentally confirmed. Nuclear PanK2 trafficked from the nucleus to the mitochondria, but not in the other direction, and was absent from the nucleus during G2 phase of the cell cycle. The localization of human PanK2 in these two compartments was in sharp contrast to mouse PanK2, which was exclusively cytosolic. These data demonstrate that PanK isoforms are differentially compartmentalized allowing them to sense CoA homeostasis in different cellular compartments and enable interaction with regulatory ligands produced in these same locations.

Support for a Fluid-continuum Model of Sexual Orientation: a Large-scale Internet Study

In a study with 17,785 subjects obtained over the Internet from the United States and 47 other countries, Kinsey's hypothesis that sexual orientation lies on a continuum was supported. Self-identifications of subjects as gay, straight, bisexual, and other corresponded to broad, skewed distributions, suggesting that such terms are misleading for many people. Sexual orientation range-roughly, how much flexibility someone has in expressing sexual orientation-was also measured. The results support a fluid-continuum model of sexual orientation, according to which genetic and environmental factors determine both the size of the sexual orientation range and the point at which an individual's sexual orientation is centered on the continuum.

Duplex Destabilization by Four Ribosomal DEAD-box Proteins

DEAD-box proteins are believed to participate in the folding of RNA by destabilizing RNA secondary or tertiary structures. Although these proteins bind and hydrolyze ATP, the mechanism for which nucleotide hydrolysis is coupled to helix destabilization may vary among different DEAD-box proteins. To investigate their abilities to disrupt helices and couple ATP hydrolysis to unwinding, the S. cerevisiae ribosomal DEAD-box proteins, Dbp3p, Dbp4p, Rok1p, and Rrp3p were assayed utilizing a series of RNA substrates containing a short duplex and either a 5' or 3' single-stranded region. All four proteins unwound a 10 base pair helix in vitro in the presence of ATP; however, significant dissociation of longer helices was not observed. While Dbp3p did not require a single-stranded extension to disrupt a helix, the unwinding activities of Dbp4p, Rok1p, and Rrp3p were substantially stimulated by either a 5' or 3' single-stranded extension. Interestingly, these proteins showed a clear length dependency with 3' extensions that was not observed with 5' extensions, suggesting that they bind substrates with a preferred orientation. In the presence of AMPPNP or ADP, all four proteins displayed displacement activity suggesting that nucleotide binding is sufficient to facilitate duplex disruption. Further enhancement of the strand displacement rate in the presence of ATP was observed only for Dbp3p and Rrp3p.

Invasion of the Spinal Column by a Posterior Mediastinal Cavernous Haemangioma: A Combined Surgical Approach

Lignopurines: A New Family of Hybrids Between Cyclolignans and Purines. Synthesis and Biological Evaluation

A new family of hybrids between cyclolignans related to podophyllic aldehyde, a non-lactonic cyclolignan, and purines were prepared and evaluated against several human tumour cell lines. Both fragments, cyclolignan and purine, were linked through aliphatic and aromatic chains. The influence on the cytotoxicity of the purine substitution and the nature of the linker is analyzed. The new family was slightly less cytotoxic than the parent podophyllic aldehyde, although the selectivity is maintained or even improved and among the linkers used, the presence of an aromatic ring gave the most potent and selective derivatives within the new series tested. Cell cycle and confocal studies demonstrate that these derivatives interfere with the tubulin polymerization and arrest cells at the G(2)/M phase, in the same way than the parent compounds podophyllotoxin and podophyllic aldehyde do.

Impact of Demographic Characteristics in Pet Ownership: Modeling Animal Count According to Owners Income and Age

Pet owner characteristics such as age, gender, income/social class, marital status, rural/urban residence and household type have been shown to be associated with the number of owned pets. However, few studies to date have attempted to evaluate these associations in Brazil. Accordingly, the aim of this study was to evaluate the association between age and income of owners and the number of owned dogs and cats in a Brazilian urban center. Pinhais, metropolitan area of Curitiba, Southern Brazil, the seventh largest city in Brazil, was chosen for this study. Questionnaires were administered door-to-door between January and February 2007 and data were analyzed by zero-inflated negative binomial (ZINB) models. A total of 13,555 of 30,380 (44.62%) households were interviewed. The majority (62.43%) of households reported having one or more dogs, with one or two dogs being the most common (29.97% and 19.71%, respectively). Cat ownership per household was much lower (P=0.001) than dog ownership, with 90% of the households reported having no owned cats. ZINB analyses indicated that income is not associated with the number of both dogs and cats among households that have pets. However, households from higher income categories were more likely to have dogs (but not cats) when compared to the lowest income category (P<0.05), contradicting a common belief that the poorer the family, the more likely they have pets. Certain age categories were significantly associated with the number of dogs or cats in households that have pets. In addition, most age categories were significantly associated with having dogs and/or cats (P<0.05). In conclusion, our study has found that age but not household income is associated with the number of dogs or cats in households that have pets; higher income households were more likely to have dogs when compared to low-income households.

Clinical Impact of RehaCom Software for Cognitive Rehabilitation of Patients with Acquired Brain Injury

We describe the clinical impact of the RehaCom computerized cognitive training program instituted in the International Neurological Restoration Center for rehabilitation of brain injury patients. Fifty patients admitted from 2008 through 2010 were trained over 60 sessions. Attention and memory functions were assessed with a pre- and post-treatment design, using the Mini-Mental State Examination, Wechsler Memory Scale and Trail Making Test (Parts A and B). Negative effects were assessed, including mental fatigue, headache and eye irritation. The program's clinical usefulness was confirmed, with 100% of patients showing improved performance in trained functions. KEYWORDS Acute brain injuries; traumatic brain injury; brain injury, vascular; rehabilitation; intellectual disability; neuropsychology; Cuba.

G-Protein Coupled Receptor Family C, Group 5, Member A (gprc5a) Expression Is Decreased in the Adjacent Field and Normal Bronchial Epithelia of Patients with Chronic Obstructive Pulmonary Disease and Non-Small-Cell Lung Cancer

: Understanding oncogenes and tumor suppressor genes expression patterns is essential for characterizing lung cancer pathogenesis. We have previously demonstrated that mGprc5a/hGPRC5A is a lung-specific tumor suppressor evidenced by inflammation-mediated tumorigenesis in Gprc5a-knockout mice. The implication of GPRC5A in human lung cancer pathogenesis, including that associated with inflammatory chronic obstructive pulmonary disease (COPD), a risk factor for the malignancy, remains elusive.

Effects of Energy Concentration of the Diet on Productive Performance and Egg Quality of Brown Egg-laying Hens Differing in Initial Body Weight

The influence of AME(n) concentration of the diet on productive performance and egg quality traits was studied in Hy-Line brown egg-laying hens differing in initial BW from 24 to 59 wk of age. Eight treatments were arranged factorially with 4 diets varying in energy content (2,650, 2,750, 2,850, and 2,950 kcal of AME(n)/kg) and 2 initial BW of the hens (1,733 vs. 1,606 g). Each treatment was replicated 5 times (13 hens per replicate), and all diets had similar nutrient content per unit of energy. No interactions between energy content of the diet and initial BW of the hens were detected for any trait. An increase in energy concentration of the diet increased (linear, P < 0.05; quadratic P < 0.05) egg production, egg mass, energy efficiency (kcal of AME(n)/g of egg), and BW gain (P < 0.05) but decreased ADFI (linear, P < 0.001) and feed conversion ratio per kilogram of eggs (linear, P < 0.01; quadratic P < 0.01). An increase in energy content of the diet reduced Haugh units and the proportion of shell in the egg (P < 0.01). Feed intake (114.6 vs. 111.1 g/hen per day), AME(n) intake (321 vs. 311 kcal/hen per day), egg weight (64.2 vs. 63.0 g), and egg mass (58.5 vs. 57.0 g) were higher for the heavier than for the lighter hens (P < 0.01), but feed conversion ratio per kilogram of eggs and energy efficiency were not affected. Eggs from the heavier hens had a higher proportion of yolk and lower proportion of albumen (P < 0.01) and shell (P < 0.05) than eggs from the lighter hens. Consequently, the yolk-to-albumen ratio was higher (P < 0.001) for the heavier hens. It is concluded that brown egg-laying hens respond with increases in egg production and egg mass to increases in AME(n) concentration of the diet up to 2,850 kcal/kg. Heavy hens had higher feed intake and produced heavier eggs and more egg mass than light hens. However, feed and energy efficiency were better for the lighter hens.

[Pleasure in Nursing Technicians Working at an Emergency Unit of a Public University Hospital]

This study aimed to reveal the main aspects of the work process and feelings of pleasure experienced by nursing technicians who work at an emergency unit in Paraná, Brazil. The theoretical basis is the psychodynamics of work. This is a qualitative and descriptive study. Data were collected and analyzed using a semi-structured interview and the content analysis technique. Subjects were selected using a snowball sampling. Important aspects of the work process were revealed such as the unpredictability of working in an emergency unit, the impact of team work, and the comprehensive care model as a precursor to humanized care. Pleasure originates from the acknowledgement of their work either by the working subject him/herself by patients or society; and from the team work, realized by the cooperation among professionals. Feelings of pleasure are linked to the acknowledgment of their work, which should be valued since gratification contributes to the psychological health of workers.

Supplemental Hydrogen Sulphide Protects Transplant Kidney Function and Prolongs Recipient Survival After Prolonged Cold Ischaemia-reperfusion Injury by Mitigating Renal Graft Apoptosis and Inflammation

What's known on the subject? and What does the study add? Hydrogen sulphide (H(2) S) has recently been classified as a member of the family of small gaseous molecules called gasotransmitters and has been found to have many important physiological functions. Several recent studies have elucidated the protective effects of H(2) S in many models of tissue ischaemia-reperfusion injury (IRI), including hepatic, myocardial, pulmonary, cerebral and renal IRI. It has previously been shown that H(2) S has a number of properties that may contribute to its protection against IRI, including vasodilatory, anti-apoptotic, anti-inflammatory and anti-oxidant effects, although the specific actions appear to vary between tissues. The few studies investigating the effects of H(2) S against renal IRI have only involved clamping of the renal pedicle to induce warm IRI. This study investigated the protective effects of H(2) S in the context of renal transplantation (RTx), which generally involves a more severe period of prolonged cold IRI. A previous study investigated the actions of H(2) S in RTx, but it was performed ex vivo and did not involve actual transplantation of donor kidneys. To our knowledge, this is the first study using a clinically relevant model of RTx to show that treatment of donor kidneys with H(2) S during preservation is protective against prolonged cold IRI. These findings suggest that H(2) S has potential utility in improving clinical organ preservation techniques and increasing the overall success of organ transplantation. OBJECTIVE: •  To characterize the effects of hydrogen sulphide (H(2) S), an endogenously produced molecule recently described to have protective effects against warm ischaemic tissue injury, in mitigating transplantation-associated prolonged cold ischaemia-reperfusion injury (IRI) in a clinically applicable in vivo model of renal transplantation (RTx). MATERIALS AND METHODS: •  After undergoing bilateral native nephrectomy, Lewis rats underwent RTx with kidneys that were flushed with either cold (4 °C) standard University of Wisconsin preservation solution (UW) or cold UW + 150 µM NaHS (H(2) S) solution and stored for 24 h at 4 °C in the same solution. •  Recipient rats were monitored for a 14-day time course using metabolic cages to assess various characteristics of renal graft function. •  Renal grafts were removed at time of death or after the rats were killed for histological, immunohistochemical and quantitative PCR analysis. RESULTS: •  H(2) S-treated rats exhibited immediate and significant (P < 0.05) decreases in serum creatinine levels, increased urine output and increased survival compared with UW-treated rats. •  H(2) S-treated grafts showed significantly reduced glomerular and tubular necrosis and apoptosis, diminished graft neutrophil and macrophage infiltrates and a trend towards improved inflammatory and anti-apoptotic cytokine profiles. CONCLUSION: •  Our results provide the first evidence that supplemental H(2) S can mitigate renal graft IRI incurred during transplantation and prolonged cold storage, improving early graft function and recipient survival in a clinically applicable model of RTx.

Defining Putative Glycan Cancer Biomarkers by MS

For decades, the association between aberrant glycosylation and many types of cancers has been shown. However, defining the changes of glycan structures has not been demonstrated until recently. This has been facilitated by the major advances in MS and separation science, which allows the detailed characterization of glycan changes associated with cancer. MS glycomics methods have been successfully employed to compare the glycomic profiles of different human specimens collected from disease-free individuals and patients with cancer. Additionally, comparing the glycomic profiles of glycoproteins purified from specimen collected from disease-free individuals and patients with cancer has also been performed. These types of glycan analyses employing MS or LC-MS allow the characterization of native, labeled and permethylated glycans. This review discusses the different glycomic and glycoproteomic methods employed for defining glycans as cancer biomarkers of different organs, including breast, colon, esophagus, liver, lung, ovarian, pancreas and prostate.

Graphene in Combination with Cucurbit[n]urils As Electrode Modifiers for Electroanalytical Biomolecules Sensing

Cucurbit[n]urils have been supported on graphene to develop sensitive and selective electrodes. The electrochemical response of modified electrodes containing graphene or graphene plus cucurbiturils has been studied for three probe molecules including hydroxymethylferrocene, ferrocyanide and methylviologen. It was found that the properties of these modified electrodes are derived from an increase in electron mobility and catalytic activity imparted by graphene and the selective complexation and molecular recognition due to cucurbit[n]urils. These properties of the graphene/cucurbit[n]urils modified electrodes have been applied for the electrochemical detection of relevant biomolecules as tryptophan at 0.69×10(-7)M concentration.

Development and Validation of a Simple and Sensitive High Performance Liquid Chromatographic Method for the Simultaneous Determination of Anastrozole, Bicalutamide, Tamoxifen, and Their Synthetic Impurities

A simple and sensitive analytical method for simultaneous determination of anastrozole, bicalutamide, and tamoxifen as well as their synthetic impurities, anastrozole pentamethyl, bicalutamide 3-fluoro-isomer, and tamoxifen e-isomer, was developed and validated by using high performance liquid chromatography (HPLC). The separation was achieved on a Symmetry(®) C-8 column (100×4.6mm i.d., 3.5μm) at room temperature (±24°C), with a mobile phase consisting of acetonitrile/water containing 0.18% N,N dimethyloctylamine and pH adjusted to 3.0 with orthophosphoric acid (46.5/53.5, v/v) at a flow rate of 1.0mLmin(-1) within 20min. The detection was made at a wavelength of 270nm by using ultraviolet (UV) detector. No interference peaks from excipients and relative retention time indicated the specificity of the method. The calibration curve showed correlation coefficients (r) >0.99 calculated by linear regression and analysis of variance (ANOVA). The limit of detection (LOD) and limit of quantitation (LOQ), respectively, were 2.2 and 6.7μgmL(-1) for anastrozole, 2.61 and 8.72μgmL(-1) for bicalutamide, 2.0 and 6.7μgmL(-1) for tamoxifen, 0.06 and 0.22μgmL(-1) for anastrozole pentamethyl, 0.02 and 0.07μgmL(-1) for bicalutamide 3-fluoro-isomer, and 0.002 and 0.007μgmL(-1) for tamoxifen e-isomer. Intraday and interday relative standard deviations (RSDs) were <2.0% (drugs) and <10% (degradation products) as well as the comparison between two different analysts, which were calculated by f test.

Intraosseous Ganglion Cyst Within the L4 Lamina Causing Spinal Stenosis

BACKGROUND CONTEXT: There are rare reports of intraosseous ganglion cysts in the cervical spine. However, to our knowledge, there are no previous reports of these cysts occurring in the lumbar spine. PURPOSE: To report a case of symptomatic lumbar spinal stenosis caused by an intraosseous ganglion cyst of the L4 lamina that communicated with the spinal canal. STUDY DESIGN: Case report. METHODS: An 86-year-old woman was referred to our spine service for a 2-year history of anterior thigh and leg pain. Magnetic resonance imaging revealed a benign-appearing intraosseous cyst in the left L4 lamina communicating with a posterior epidural cyst at L4-L5 causing marked spinal stenosis. The patient was treated successfully with a laminectomy and resection. RESULTS: The patient underwent partial laminectomies of L4 and L5 preserving the interspinous ligaments between L5-S1 and L3-L4. The cyst was removed en bloc without violation of the cyst wall. Histopathologic examination revealed focal myxoid changes without a cellular lining of the cyst wall, confirming the diagnosis of intraosseous ganglion cyst. CONCLUSIONS: This is the first report to describe an intraosseous ganglion cyst occurring in the lumbar spine. Although spinal stenosis is commonly a result of degenerative joint or disc disease, it occasionally may result from more obscure causes. This case illustrates a patient with an intraosseous ganglion cyst within the spinal lamina resulting in spinal stenosis, treated successfully with a laminectomy and resection.

Dissecting T Cell Contraction In Vivo Using a Genetically Encoded Reporter of Apoptosis

Contraction is a critical phase of immunity whereby the vast majority of effector T cells die by apoptosis, sparing a population of long-lived memory cells. Where, when, and why contraction occurs has been difficult to address directly due in large part to the rapid clearance of apoptotic T cells in vivo. To circumvent this issue, we introduced a genetically encoded reporter for caspase-3 activity into naive T cells to identify cells entering the contraction phase. Using two-photon imaging, we found that caspase-3 activity in T cells was maximal at the peak of the response and was associated with loss of motility followed minutes later by cell death. We demonstrated that contraction is a widespread process occurring uniformly in all organs tested and targeting phenotypically diverse T cells. Importantly, we identified a critical window of time during which antigen encounters act to antagonize T cell apoptosis, supporting a causal link between antigen clearance and T cell contraction. Our results offer insight into a poorly explored phase of immunity and provide a versatile methodology to study apoptosis during the development or function of a variety of immune cells in vivo.

Cerebral Desaturation Events in the Intensive Care Unit Following Cardiac Surgery

PURPOSE: Patients may be at high risk for hemodynamic instability in the early postoperative period, with subsequent poor cerebral perfusion and the development of postoperative cerebral oxygen desaturation events (CDEs). Intraoperative CDEs have been associated with postoperative adverse events. However, none of these studies examined the incidence of early postoperative cerebral desaturations. This study was designed to identify the incidence of CDEs (defined as a decrease in SctO(2) to less than 60% for at least 60 seconds) in the immediate postoperative period following cardiac surgery. METHODS: Fifty-three moderate to high-risk patients undergoing elective cardiac surgery were enrolled in this observational study. Cerebral oximeter monitors were placed on all patients prior to induction of anesthesia and remained in place for 6 hours or until the patients were extubated postoperatively, whichever occurred first. Data were recorded from the cerebral oximeter, physiologic monitor and ventilator during the study period. Data were analyzed to identify the incidence of early postoperative CDEs, as well as association with subsequent clinical events. RESULTS: The incidence of early postoperative CDEs was 53%. Sixty-four percent of these CDEs lasted for more than 1 hour. Patients with postoperative CDEs were more likely to have had intraoperative CDEs (P< 0.0001). Five out of 28 patients who experienced CDEs in the intensive care unit died while none of the patients without postoperative CDEs died (P = .053). CONCLUSION: A high incidence of CDEs (53%) was found in the early post-cardiac surgery period. Larger studies are needed to determine whether postoperative CDEs are correlated with various physiologic events or are associated with adverse patient outcomes.

TLR Agonists Are Highly Effective at Eliciting Functional Memory CTLs of Effector Memory Phenotype in Peptide Immunization

Given the importance of memory cytotoxic T lymphocytes (CTLs) in eliminating altered self-cells, including virus-infected and tumor cells, devising effective vaccination strategies for generating memory CTLs is a priority in the field of immunology. Herein, we elaborate upon a novel boosting approach that utilizes synthetic peptides and Toll-like receptor (TLR) agonists as adjuvants to generate sufficient numbers of memory CTLs to protect against infection in mice. Peptide boosting with lipopolysaccharide (LPS), a TLR4-ligand, has been shown to progressively enhance memory CTLs. Whether this result is strictly dependent on activation of TLR4 or can be similarly achieved by signaling through other TLRs is of practical interest in vaccine development but is yet unknown. In this report, we present evidence that intravenous peptide boosting together with TLR3 and TLR9 agonists (Poly IC and CpG, respectively) is highly effective and induces large quantities of memory CTLs of effector memory phenotype after three boosts. Compared to LPS, CpG and Poly IC generate more robust immune responses after the first and second boosts, indicating that a protective level of CTLs might be achieved with fewer boosts when CpG or Poly IC is used. Lastly, the resultant memory CTLs from boosting with different TLR agonists as adjuvant are equally protective against pathogen challenge and are not immune senescent. Therefore, TLR agonists are effective adjuvants in intravenous peptide boosting for the generation of functional memory CTLs.

Seizure Exacerbation in Two Patients with Focal Epilepsy Following Marijuana Cessation

While animal models of epilepsy suggest that exogenous cannabinoids may have anticonvulsant properties, scant evidence exists for these compounds' efficacy in humans. Here, we report on two patients whose focal epilepsy was nearly controlled through regular outpatient marijuana use. Both stopped marijuana upon admission to our epilepsy monitoring unit (EMU) and developed a dramatic increase in seizure frequency documented by video-EEG telemetry. These seizures occurred in the absence of other provocative procedures, including changes to anticonvulsant medications. We review these cases and discuss mechanisms for the potentially anticonvulsant properties of cannabis, based on a review of the literature.

Early Alterations in Plasma Ghrelin Levels in Offspring of Calorie-restricted Rats During Gestation May Be Linked to Lower Sympathetic Drive to the Stomach

Serum ghrelin concentration is generally reduced in obesity. We aimed to assess whether this alteration is present in rats predisposed to obesity because of moderate undernutrition during gestation, and to explore whether this could be related with alterations in stomach sympathetic innervation, which is involved in gastric ghrelin secretion. Offspring of control and 20% gestational calorie-restricted dams (CR) exposed to normal-fat-diet from weaning onward were studied. Circulating ghrelin levels were measured at 25 days and 4 months of age. Morphometry, number of ghrelin-positive (ghrelin(+)) cells, ghrelin mRNA and protein levels, and tyrosine hydroxylase (TH) protein levels in stomach were determined at 25 days. Adult CR male animals, but not females, exhibited greater body-weight (BW) than their controls, but both males and females showed lower circulating ghrelin levels. This alteration in ghrelin levels was already present at 25 days, prior to any difference in BW. At this juvenile age, no differences in gastric morphometry, number of ghrelin(+) cells or ghrelin mRNA/protein levels were found between control and CR animals, however, CR animals showed lower TH stomach content. These results suggest that circulating ghrelin concentration is early altered in rats prenatally programmed to develop obesity. This does not seem to be associated with lower ghrelin production capacity but with specific alterations in sympathetic drive to the stomach.

VE-Cadherin Signaling Induces EB3 Phosphorylation to Suppress Microtubule Growth and Assemble Adherens Junctions

Vascular endothelial (VE)-cadherin homophilic adhesion controls endothelial barrier permeability through assembly of adherens junctions (AJs). We observed that loss of VE-cadherin-mediated adhesion induced the activation of Src and phospholipase C (PLC)γ2, which mediated Ca(2+) release from endoplasmic reticulum (ER) stores, resulting in activation of calcineurin (CaN), a Ca(2+)-dependent phosphatase. Downregulation of CaN activity induced phosphorylation of serine 162 in end binding (EB) protein 3. This phospho-switch was required to destabilize the EB3 dimer, suppress microtubule (MT) growth, and assemble AJs. The phospho-defective S162A EB3 mutant, in contrast, induced MT growth in confluent endothelial monolayers and disassembled AJs. Thus, VE-cadherin outside-in signaling regulates cytosolic Ca(2+) homeostasis and EB3 phosphorylation, which are required for assembly of AJs. These results identify a pivotal function of VE-cadherin homophilic interaction in modulating endothelial barrier through the tuning of MT dynamics.

Candidate Gene Association Study of BMI-Related Loci, Weight, and Adiposity in Old Age

Most genome-wide association studies are confined to middle-aged populations. It is unclear whether associations between single nucleotide polymorphisms (SNPs) and obesity persist in old age. We aimed to relate 10 body mass index (BMI)-associated SNPs to weight, BMI, % fat, visceral and subcutaneous adipose tissue in Health ABC and AGES-Reykjavik comprising 4,846 individuals of European Ancestry, and 1,139 African Americans over age 65. SNPs were scaled using effect estimates from candidate SNPs. In Health ABC, a SNP near GNPDA2 was modestly associated with weight and SAT area (p = .008, p = .001). Risk score (sum of scaled SNPs) was associated with weight, BMI, and SAT area (p < .0001 for all), but neither GNPDA2 nor risk score was associated with weight, BMI, visceral adippose tissue, subcutaneous adipose tissue, or % fat in AGES-Reykjavik. In African Americans, a SNP near SEC16B was weakly associated with weight (p = .04). In this sample of older adults, no BMI-associated SNPs were associated with weight or adiposity.

Twist Buckling Behavior of Arteries

Arteries are often subjected to torsion due to body movement and surgical procedures. While it is essential that arteries remain stable and patent under twisting loads, the stability of arteries under torsion is poorly understood. The goal of this work was to experimentally investigate the buckling behavior of arteries under torsion and to determine the critical buckling torque, the critical buckling twist angle, and the buckling shape. Porcine common carotid arteries were slowly twisted in vitro until buckling occurred while subjected to a constant axial stretch ratio (1.1, 1.3, 1.5 (in vivo level) and 1.7) and lumen pressure (20, 40, 70 and 100 mmHg). Upon buckling, the arteries snapped to form a kink. For a group of six arteries, the axial stretch ratio significantly affected the critical buckling torque ([Formula: see text]) and the critical buckling twist angle ([Formula: see text]). Lumen pressure also significantly affected the critical buckling torque ([Formula: see text]) but had no significant effect on the critical twist angle ([Formula: see text]). Convex material constants for a Fung strain energy function were determined and fit well with the axial force, lumen pressure, and torque data measured pre-buckling. The material constants are valid for axial stretch ratios, lumen pressures, and rotation angles of 1.3-1.5, 20-100 mmHg, and 0-270[Formula: see text], respectively. The current study elucidates the buckling behavior of arteries under torsion and provides new insight into mechanical instability of blood vessels.

Coordinated C-di-GMP Repression of Salmonella Motility Through YcgR and Cellulose

Cyclic di-GMP (c-di-GMP) is a secondary messenger that controls a variety of cellular processes including the switch between a biofilm and a planktonic bacterial lifestyle. This nucleotide binds to cellular effectors in order to exert its regulatory functions. In Salmonella, two proteins, BcsA and YcgR, both of them containing a c-di-GMP binding PilZ domain, are the only known c-di-GMP receptors. BcsA, upon c-di-GMP binding, synthesizes cellulose, the main exopolysaccharide of the biofilm matrix. YcgR is dedicated to c-di-GMP dependent inhibition of motility through its interaction with flagellar motor proteins. However, previous evidences indicate that in the absence of YcgR there is still an additional element that mediates motility impairment under high c-di-GMP levels. Here, we uncover that cellulose per se is the factor that further promotes inhibition of bacterial motility once high c-di-GMP contents drive the activation of a sessile lifestyle. Inactivation of different genes of the bcsABZC operon, mutation of the conserved residues in the RxxxR motif of the BcsA PilZ domain or degradation of the cellulose produced by BcsA rescued the motility defect of ΔycgR strains in which high c-di-GMP levels were reached through the overexpression of diguanylate cyclases. High c-di-GMP levels provoked cellulose accumulation around cells that impeded flagella rotation probably by means of steric hindrance, without affecting flagella gene expression, exportation or assembly. Our results highlight the relevance of cellulose in Salmonella lifestyle switching as an architectural element that is both essential for biofilm development and required, in collaboration with YcgR, for complete motility inhibition.

The Roles of Voltage-gated Potassium Channels Kv2.1 and Kv2.2 in the Regulation of Insulin and Somatostatin Release from Pancreatic Islets

The voltage-gated potassium channels Kv2.1 and Kv2.2 are highly expressed in pancreatic islets, yet their contribution to islet hormone secretion are not fully understood. Here we investigate the role of Kv2 channels in pancreatic islets using a combination of genetic and pharmacological approaches. Pancreatic β-cells from Kv2.1(-/-) mice possess reduced Kv current and display greater glucose-stimulated insulin secretion (GSIS) relative to wild-type β-cells. Inhibition of Kv2.x channels with selective peptidyl (GxTX-1E) or small molecule (RY796) inhibitors enhances GSIS in isolated wild-type mouse and human islets, but not in islets from Kv2.1(-/-) mice. However, in wild-type mice neither inhibitor improved glucose tolerance in vivo. GxTX-1E and RY796 enhanced somatostatin release in isolated human and mouse islets and in situ perfused pancreata from wild-type and Kv2.1-/- mice. Kv2.2 silencing in mouse islets by adenovirus-shRNA specifically enhanced islet somatostatin, but not insulin, secretion. In mice lacking somatostatin receptor 5, GxTX-1E stimulated insulin secretion and improved glucose tolerance. Collectively, these data show that Kv2.1 regulates insulin secretion in β-cells and Kv2.2 modulates somatostatin release in δ-cells. Development of selective Kv2.1 inhibitors without cross inhibition of Kv2.2 may provide new avenues to promote GSIS for the treatment of type 2 diabetes.

In-depth Proteomic Analysis of Banana (Musa Spp.) Fruit with Combinatorial Peptide Ligand Libraries

Musa ssp. is among the world leading fruit crops. Although a strong interest on banana biochemistry exists in the scientific community, focused on metabolite composition, proteins have been scarcely investigated even if they play an important role in food allergy and stability, are a source of biologically active peptides and can provide information about nutritional aspects of this fruit. In this work we have employed the combinatorial peptide ligand libraries after different types of protein extractions, for searching the very-low abundance proteins in banana. The use of advanced mass spectrometry techniques and musa mRNAs database in combination with the Uniprot_viridiplantae database allowed us to identify 1131 proteins. Among this huge amount of proteins we found several already known allergens such as musa a 1, pectinesterase, superoxide dismutase and potentially new allergens. Additionally several enzymes involved in degradation of starch granules and strictly correlated to ripening stage were identified. This is the first in depth exploration of the banana fruit proteome and one of the largest descriptions of the proteome of any vegetable system.

Invasive Aspergillosis Caused by Cryptic Aspergillus Species: a Report of Two Consecutive Episodes in a Patient with Leukemia

We report a case of two consecutive episodes of invasive aspergillosis caused by cryptic Aspergillus species in a patient with leukemia. A first episode of pulmonary infection was caused by Aspergillus calidoustus and A. novofumigatus, and the second episode by A. novofumigatus and A. viridinutans. Fungal isolates were identified to species level using traditional and sequencing-based molecular methods.

Interaction of Biphenyl-Functionalized Eu(2+)-Containing Cryptate with Albumin: Implications to Contrast Agents in Magnetic Resonance Imaging

The influence of albumin on the efficacy of a Eu(2+)-containing complex capable of interacting with human serum albumin (HSA) was investigated at different field strengths (1.4, 3, 7, 9.4, and 11.7 T). Relaxometric measurements indicated that the presence of albumin at higher field strengths (>3 T) did not result in an increase in the relaxivity of the Eu(2+) complex, but a relaxation enhancement of 171 ± 11% was observed at 1.4 T. Titration experiments using different percentages (2, 4.5, 6, 10, 15, and 25% w/v) of HSA and variable-temperature (17)O NMR measurements were performed to understand the effect of albumin on the molecular properties of the biphenyl-functionalized Eu(2+) complex that are relevant to magnetic resonance imaging.

Effectiveness of the Bevacizumab-irinotecan Regimen in the Treatment of Recurrent Glioblastoma Multiforme: Comparison with Other Second-line Treatments Without This Regimen

A retrospective cohort study was conducted to analyse the effectiveness of bevacizumab and irinotecan (BVZ/CPT-11) as a second-line treatment in patients with primary glioblastoma multiforme (GBM) in comparison with a control group that were not administered BVZ/CPT-11 at the first recurrence. The difference in overall survival (OS) between the two groups was used as a predictor of effectiveness. OS was calculated according to prognostic factors and gender. A total of 28 and 32 patients were enrolled in the BVZ/CPT-11 cohort and control group, respectively. The median OS was 17.94 months (95% CI, 14.91-20.96) in the BVZ/CPT-11 treatment cohort and 10.97 months (95% CI, 7.65-14.30) in the control cohort. The results obtained on the effectiveness of BVZ/CPT-11 treatment in patients with primary GBM are consistent with data from previous studies. No significant differences were identified in OS based on prognostic factors; therefore, the latter cannot be used to select patients who would incur the greatest benefits from BVZ/CPT-11 treatment.

Plasma Malondialdehyde Levels and Risk Factors for the Development of Chronic Complications in Type 2 Diabetic Patients on Insulin Therapy

We assessed plasma malondialdehyde (MDA) levels as a biomarker of lipid peroxidation in type 2 diabetic patients on insulin therapy. Associations among MDA levels and some risk factors for the development of chronic complications of diabetes were also evaluated.

Identification and Virulence of Chryseobacterium Indologenes Isolated from Diseased Yellow Perch (Perca Flavescens)

AIM: To identify pathogen of diseased yellow perch and determine their virulence. METHODS AND RESULTS: Fifteen Gram-negative bacteria isolates were recovered from the skin lesions of diseased yellow perch (Perca flavescens). Based on API 20NE test, ten isolates were found to share 67.2% to 99.9% homologies with Chryseobactertium indologenes. Based on fatty acid methyl ester analysis, thirteen isolates were found to share similarities with C. indologenes and other species of Chryseobacterium. Based on sequencing results of partial 16S rRNA gene, thirteen isolates shared 99% identities (e value = 2e-50) with the 16S rRNA sequence of C. indologenes (GenBank HQ259684). Based on the 16S-23S rRNA intergenic spacer region (ISR) sequence, the thirteen isolates shared 88% identity (e value = 1e-165) with the 16S-23S ISR sequence of C. indologenes (GenBank EU014570). T-coffee multiple sequence alignment revealed that the partial 16S rRNA or the 16S-23S ISR sequence of the thirteen isolates shared 100% identity with each other. When healthy yellow perch were exposed to the fifteen isolates by bath immersion (~ 6 × 10(7) CFU/ml for one hour), only C. indologenes isolates killed 10% to 20% of fish, whereas other isolates were avirulent. When yellow perch were exposed to C. indologenes by intraperitoneal injection, mortality was dose-dependent, with LD(50) and LD(95) values of 1.5 × 10(8) and 3.2 × 10(8) CFU/fish, respectively. CONCLUSIONS: C. indologenes could be pathogenic to yellow perch. © No claim to US Government works. Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

Generation of Inner Ear Sensory Cells from Bone Marrow-derived Human Mesenchymal Stem Cells

Aim: Hearing loss is the most common sensory disorder in humans, its main cause being the loss of cochlear hair cells. We studied the potential of human mesenchymal stem cells (hMSCs) to differentiate towards hair cells and auditory neurons. Materials & methods: hMSCs were first differentiated to neural progenitors and subsequently to hair cell- or auditory neuron-like cells using in vitro culture methods. Results: Differentiation of hMSCs to an intermediate neural progenitor stage was critical for obtaining inner ear sensory lineages. hMSCs generated hair cell-like cells only when neural progenitors derived from nonadherent hMSC cultures grown in serum-free medium were exposed to EGF and retinoic acid. Auditory neuron-like cells were obtained when treated with retinoic acid, and in the presence of defined growth factor combinations containing Sonic Hedgehog. Conclusion: The results show the potential of hMSCs to give rise to inner ear sensory cells.

Strategies for Monitoring and Updating Clinical Practice Guidelines: a Systematic Review

ABSTRACT: BACKGROUND: Scientific knowledge is in constant change. The flow of new information requires a frequent re-evaluation of the available research results. Clinical practice guidelines (CPGs) are not exempted from this phenomenon and need to be kept updated to maintain the validity of their recommendations. The objective of the study is to systematically identify, describe and assess strategies for monitoring and updating CPGs. Study design and setting We conducted a systematic review of studies evaluating one or more methods of updating (with or without monitoring) CPGs or recommendations. We searched MEDLINE (PubMed) and The Cochrane Methodology Register (The Cochrane Library) from 1966 to June 2012. Additionally, we hand-searched reference lists of the included studies and the Guidelines International Network book of abstracts. If necessary, we contacted study authors to obtain additional information. RESULTS: We included a total of eight studies. Four evaluated if CPGs were out of date, three updated CPGs, and one continuously monitored and updated CPGs. The most detailed phase of the process reported was the identification of new evidence. As opposed to studies updating guidelines, studies evaluating if CPGs were out of date applied restricted searches. Only one study compared a restricted versus an exhaustive search suggesting that a restricted search is sufficient to assess recommendations' recency. One study analyzed the survival time of CPGs and suggested that these should be reassessed every three years. CONCLUSIONS: There is limited evidence about the optimal strategies for monitoring and updating clinical practice guidelines. A restricted search is likely to be sufficient to monitor new evidence and assess the need to update; however, more information is needed about the timing and type of search. Only the exhaustive search strategy has been assessed for the update of CPGs. The development and evaluation of more efficient strategies is needed to improve the timeliness and reduce the burden of maintaining the validity of CPGs.

Short Communication: The Effect of Premilking with a Teat Cup-like Device, in a Novel Robotic Rotary, on Attachment Accuracy and Milk Removal

This study investigated the effects of premilking teat preparation on attachment accuracy and milk removal characteristics for individual cows in a novel 16-bail prototype robotic rotary (RR; automatic milking rotary system; DeLaval AMR, Tumba, Sweden). The first commercial versions of the RR systems will have the option of purchasing and installing a teat preparation module (TPM) for premilking stimulation and cleaning of teats. It is expected that attachment of teat cups would be faster and more successful with the use of a TPM, and that the efficiency of milk removal, in terms of average and peak milk flow rates, would increase. We observed a significant effect of treatment (no wash vs. wash) and individual quarters on attachment success: cows exposed to the wash treatment had up to 1.5 times higher odds of being successfully attached. The attachment was not only more successful but was also found to be 4.3 s faster after cows were exposed to the wash treatment. Average milk flow rate was not affected by the wash treatment. Nevertheless, a significant interaction was found between wash treatment and milking interval affecting the peak milk flow (kg/min) of individual cows. This interaction showed that cows with a milking interval ≤8 h subjected to the wash treatment had significantly higher peak flow rates (300 g/min increase) compared with cows in the same milking interval category with no wash treatment. The relationship between premilking stimulation and attachment success shown in this study will increase awareness (of both farmers and developers of the technology) of the importance of teat cleaning within the RR. The effects of the improved system performance should be taken into account (alongside the capital investment cost) when deciding to install a RR equipped (or not) with a TPM.

[The Usefulness of Densitometry As a Method of Assessing the Nutritional Status of Athletes. Comparison with Body Mass Index.]

The body mass index (BMI) is used to assess nutritional status. The result in athletes may be overestimated due to increase in muscle mass. OBJECTIVE: To assess the usefulness of fat mass index (FMI) and lean mass index (LMI) determination as indicators of nutritional status and to compare the results with BMI. MATERIAL AND METHODS: We studied 28 amateur rugby players, male. After being subjected to whole body densitometry by dual X-ray absorptiometry, we determined fat and lean body mass together with other parameters. FMI (fat in kg/height in meters(2)), LMI (lean in kg/height in meters(2)) and appendicular muscle mass index (AMMI, arms and legs musculature in kg/height in meters(2)) were calculated. RESULTS: Using BMI, 18 players were overweight and 4 obese type I. Considering FMI, 7 of them had normal values and high LMI and AMMI, one of them changed from overweight to obese and another one from obese to overweight. Of the 6 players with normal BMI, one of them showed fat excess and another one fat defect. The results changed the assessment of nutritional status in 39% of players. CONCLUSIONS: Although BMI is an appropriate parameter in general population for the assessment of nutritional status, in athletes should be taken into account fat and muscle body percentage and their corresponding indexes. The whole body densitometry appears to be a simple and reliable technique for this purpose.

[Impact of Quality Improvement Process Upon the State of Nutritional Support in a Critical Care Unit]

Introduction: In a preceding article the state of Nutritional support (NS) in an Intensive Care Unit (ICU) was documented [Martinuzzi A et al. Estado del soporte nutricional en una unidad de Cuidados críticos. RNC 2011; 20: 5-17]. In this follow-up work we set to assess the impact of several organizational, recording and educational interventions upon the current state of NS processes. Materials and methods: Interventions comprised presentation of the results of the audit conducted at the ICU before the institution's medical as well as paramedical personnel; their publication in a periodical, peerreviewed journal; drafting and implementation of a protocol regulating NS schemes to be carried out at the ICU; and conduction of continuous education activities on Nutrition (such as "experts talks", interactive courses, and training in the implementation of the NS protocol). The state of NS processes documented after the interventions was compared with the results annotated in the preceding article. Study observation window ran between March the 1st, 2011 and May 31th, 2011, both included. Results: Study series differed only regarding overallmortality: Phase 1: 40.0% vs. Phase 2: 20.5%; Difference: 19.5%; Z = 1.927; two-tailed-p = 0.054. Interventions resulted in a higher fulfillment rate of the prescribed NS indication; an increase in the number of patients receiving ≥ 80% of prescribed energy; and a reduction in the number of NS lost days. Mortality was (numerically) lower in patients in which the prescribed NS scheme was fulfilled, NS was early initiated, and whom received ≥ 80% of prescribed energy. Adopted interventions had no effect upon average energy intakes: Phase 1: 574.7 ± 395.3 kcal/24 h-1 vs. Phase 2: 591.1 ± 315.3 kcal/24 h¹; two-tailed-p > 0.05. Conclusions: Educational, recording and organizational interventions might result in a better conduction of NS processes, and thus, in a lower mortality. Hemodynamic instability is still the most formidable obstacle for initiating and completing NS.

First Paramagnetic Pd(II) Complex with a PdN(4)S(2) Coordination Core

Reaction of the potassium salt of N-thiophosphorylated thiourea 2-PyNHC(S)NHP(S)(OiPr)(2) (HL) with PdCl(2) in aqueous MeOH leads to the formation of two mononuclear homoleptic complexes [Pd{2-PyNHC(S)NP(S)(OiPr)(2)-1,5-S,S'}(2)] ([]) and [Pd{2-PyNHC(S)NP(S)(OiPr)(2)-1,5,7-N,N',S}(2)] ([]), while the mechanically induced solid-state reaction of KL and PdCl(2) leads to the exclusive formation of []. The dithiocoordinated desmotrope is square-planar and diamagnetic, while the second desmotrope is unprecedentedly octahedrally configured and paramagnetic.

The Human Microbiome Project Strategy for Comprehensive Sampling of the Human Microbiome and Why It Matters

The Human Microbiome Project used rigorous good clinical practice standards to complete comprehensive body site sampling in healthy 18- to 40-yr-old adults, creating an unparalleled reference set of microbiome specimens. To ensure that specimens represented minimally perturbed microbiomes, we first screened potential participants using exclusion criteria based on health history, including the presence of systemic diseases (e.g., hypertension, cancer, or immunodeficiency or autoimmune disorders), use of potential immunomodulators, and recent use of antibiotics or probiotics. Subsequent physical examinations excluded individuals based on body mass index (BMI), cutaneous lesions, and oral health. We screened 554 individuals to enroll 300 (149 men and 151 women, mean age 26 yr, mean BMI 24 kg/m(2), 20.0% racial minority, and 10.7% Hispanic). We obtained specimens from the oral cavity, nares, skin, gastrointestinal tract, and vagina (15 specimens from men and 18 from women). The study evaluated longitudinal changes in an individual's microbiome by sampling 279 participants twice (mean 212 d after the first sampling; range 30-359 d) and 100 individuals 3 times (mean 72 d after the second sampling; range 30-224 d). This sampling strategy yielded 11,174 primary specimens, from which 12,479 DNA samples were submitted to 4 centers for metagenomic sequencing. Our clinical design and well-defined reference cohort has laid a foundation for microbiome research.-Aagaard, K., Petrosino, J., Keitel, W., Watson, M., Katancik, J., Garcia, N., Patel, S., Cutting, M., Madden, T., Hamilton, H., Harris, E., Gevers, D., Simone, G., McInnes, P., Versalovic, J. The Human Microbiome Project strategy for comprehensive sampling of the human microbiome and why it matters.

C. Elegans Dopaminergic D2-Like Receptors Delimit Recurrent Cholinergic-Mediated Motor Programs During a Goal-Oriented Behavior

Caenorhabditis elegans male copulation requires coordinated temporal-spatial execution of different motor outputs. During mating, a cloacal circuit consisting of cholinergic sensory-motor neurons and sex muscles maintains the male's position and executes copulatory spicule thrusts at his mate's vulva. However, distinct signaling mechanisms that delimit these behaviors to their proper context are unclear. We found that dopamine (DA) signaling directs copulatory spicule insertion attempts to the hermaphrodite vulva by dampening spurious stimulus-independent sex muscle contractions. From pharmacology and genetic analyses, DA antagonizes stimulatory ACh signaling via the D2-like receptors, DOP-2 and DOP-3, and Gα(o/i) proteins, GOA-1 and GPA-7. Calcium imaging and optogenetics suggest that heightened DA-expressing ray neuron activities coincide with the cholinergic cloacal ganglia function during spicule insertion attempts. D2-like receptor signaling also attenuates the excitability of additional mating circuits to reduce the duration of mating attempts with unproductive and/or inappropriate partners. This suggests that, during wild-type mating, simultaneous DA-ACh signaling modulates the activity threshold of repetitive motor programs, thus confining the behavior to the proper situational context.

Conformational Changes of Blood ACE in Chronic Uremia

The pattern of binding of monoclonal antibodies (mAbs) to 16 epitopes on human angiotensin I-converting enzyme (ACE) comprise a conformational ACE fingerprint and is a sensitive marker of subtle protein conformational changes.

Hyperbaric Oxygen Therapy Treatment for the Fixation of Implant Prosthesis in Oncology Patients Irradiated

doi: 10.1111/j.1741-2358.2012.00636.x Hyperbaric oxygen therapy treatment for the fixation of implant prosthesis in oncology patients irradiated Objectives:  This study aimed to present a clinical report of an irradiated oncologic patient who underwent hyperbaric oxygen therapy to be rehabilitated with implant-supported prosthesis. Materials and Methods:  A 67-year-old man was admitted at Oral Oncology Center (FOA-UNESP) presenting a lesion on the mouth floor. After clinical evaluation, incisional biopsy and histopathological exam, a grade II squamous cell carcinoma was diagnosed. The patient was subjected to surgery to remove the lesion and partial glossectomy. Afterwards, the radiotherapy, in the left/right cervicofacial area of the supraclavicular fossa, was conducted. After 3 years of the surgery, the patient was submitted to hyperbaric oxygen therapy. Then, four implants were installed in patient's mandible. Five months later, an upper conventional complete denture and lower full-arch implant-supported prosthesis were fabricated. Conclusion:  The treatment resulted in several benefits such as improving his chewing efficiency, swallowing and speech, less denture trauma on the mucosa and improving his self-esteem.

Equinatoxin II Potentiates Temozolomide- And Etoposide-Induced Glioblastoma Cell Death

Glioblastoma (GBM) is considered incurable due to its resistance to current cancer treatments. So far, all clinically available alternatives for treating GBM are limited, evoking the development of novel treatment strategies that can more effectively manage these tumors. Extensive effort is being dedicated to characterize the molecular basis of GBM resistance to chemotherapy and to explore novel therapeutic procedures that may improve overall survival. Cytolysins are toxins that form pores in target cell membranes, modifying ion homeostasis and leading to cell death. These pore-forming toxins might be used, therefore, to enhance the efficiency of conventional chemotherapeutic drugs, facilitating their entrance into the cell. In this study, we show that a non-cytotoxic concentration of equinatoxin II (EqTx-II), a pore-forming toxin from the sea anemone Actinia equina, potentiates the cytotoxicity induced by temozolomide (TMZ), a first-line GBM treatment, and to etoposide (VP-16), a second- or third-line GBM treatment. We also suggest that this effect is selective to GBM cells and occurs via PI3K/Akt pathway inhibition. Finally, Magnetic resonance imaging (MRI) revealed that a non-cytotoxic concentration of EqTx-II potentiates the VP-16-induced inhibition of GBM growth in vivo. These combination therapies constitute a new and potentially valuable tool for GBM treatment, leading to the requirement of lower concentrations of chemotherapeutic drugs and possibly reducing, therefore, the adverse effects of chemotherapy.

BAMBI (BMP and Activin Membrane-bound Inhibitor) Protects the Murine Heart from Pressure-overload Biomechanical Stress by Restraining TGF-β Signaling

Left ventricular (LV) pressure overload is a major cause of heart failure. Transforming growth factors-β (TGF-βs) promote LV remodeling under biomechanical stress. BAMBI (BMP and activin membrane-bound inhibitor) is a pseudoreceptor that negatively modulates TGF-β signaling. The present study tests the hypothesis that BAMBI plays a protective role during the adverse LV remodeling under pressure overload. The subjects of the study were BAMBI knockout mice (BAMBI(-/-)) undergoing transverse aortic constriction (TAC) and patients with severe aortic stenosis (AS). We examined LV gene and protein expression of remodeling-related elements, histological fibrosis, and heart morphology and function. LV expression of BAMBI was increased in AS patients and TAC-mice and correlated directly with TGF-β. BAMBI deletion led to a gain of myocardial TGF-β signaling through canonical (Smads) and non-canonical (TAK1-p38 and TAK1-JNK) pathways. As a consequence, the remodeling response to pressure overload in BAMBI(-/-) mice was exacerbated in terms of hypertrophy, chamber dilation, deterioration of long-axis LV systolic function and diastolic dysfunction. Functional remodeling associated transcriptional activation of fibrosis-related TGF-β targets, up-regulation of the profibrotic micro-RNA-21, histological fibrosis and increased metalloproteinase-2 activity. Histological remodeling in BAMBI(-/-) mice involved TGF-βs. BAMBI deletion in primary cardiac fibroblasts exacerbated TGF-β-induced profibrotic responses while BAMBI overexpression in NIH-3T3 fibroblasts attenuated them. Our findings identify BAMBI as a critical negative modulator of myocardial remodeling under pressure overload. We suggest that BAMBI is involved in negative feedback loops that restrain the TGF-β remodeling signals to protect the pressure-overloaded myocardium from uncontrolled extracellular matrix deposition in humans and mice.

A New Pentaplex-nested PCR to Detect Five Pathogenic Bacteria in Free Living Amoebae

Changes in water use and anthropogenic activity have major impacts on the quality of natural aquatic ecosystems, water distribution and wastewater plants. One of the main problems is the presence of some pathogenic microorganisms that are resistant to disinfection procedures when they are hosted by free living amoeba and that in many cases are hardly detectable by culture-based procedures. In this work we report a sensitive, low-cost procedure consisting of a pentaplex-nested PCR that allows simultaneous detection of Legionella pneumophila, Mycobacterium spp., Pseudomonas spp., Vibrio cholerae and the microcystin-producing cyanobacteria Microcystis aeruginosa. The method has been used to detect the presence of these pathogenic bacteria in water and inside free living amoeba. Its validation in 72 samples obtained from different water sources from Aragon (Spain) evidences that Mycobacterium and Pseudomonas spp are prevailing as amoeba-resistant bacteria.

Low Hepatitis B Envelope Antigen Seroconversion Rate in Chronic Hepatitis B Patients on Long-term Entecavir 0.5 Mg Daily in Routine Clinical Practice

AIM: Data from registration trials with highly selective patients have shown that hepatitis B envelope antigen (HBeAg)-positive patients with chronic hepatitis B respond well to entecavir (ETV) 0.5 mg daily, with an HBeAg seroconversion rate of 21% at 12 months. However, there are varying data on the treatment outcomes of ETV 0.5 mg daily in routine clinical settings, with seroconversion rates at 12 months ranging from 8 to 48% in studies limited to 44-90 patients from centers in Asia, Europe, and South America. MATERIALS AND METHODS: In the present study, we examined long-term treatment efficacy and tolerability in 136 consecutive treatment-naive HBeAg-positive chronic hepatitis B patients treated between January 2005 and January 2011 with ETV 0.5 mg daily at community clinics and tertiary centers in the USA. The primary study end point was HBeAg seroconversion. RESULTS: Sixty-one percent of HBeAg-positive patients were men, mean age 39±12 years, median hepatitis B virus DNA 7.48 (3.7-9.8) log10 IU/ml, median alanine aminotransferase 67 (14-1077) U/l, and median treatment duration 18 (6-60) months. At months 12, 24, and 36, complete viral suppression rates were 41, 66, and 85% and HBeAg seroconversion rates were 4.8, 20, and 30%, respectively. No patients experienced adverse events or developed genotypic resistance to ETV. CONCLUSION: In clinical settings, ETV is highly tolerable and potent at suppressing hepatitis B viremia; however, the rates of HBeAg seroconversion appear to be much lower than those reported, highlighting the importance of appropriate counseling and planning for long-term therapy.

Microfiltration Applied to Dairy Streams: Removal of Bacteria

Microfiltration applied in the dairy industry for bacteria removal is an important technology for extending the shelf life of milk while maintaining or even improving its organoleptic and nutritional properties. This article reviews the evolution of this technique over recent years and the advances currently being made in the field. The cited literature indicates the strategies used to overcome the main drawbacks of this type of operation, the most common operating conditions employed and the reduction degree of bacteria obtained. © 2012 Society of Chemical Industry.

Host Cell Factors As Antiviral Targets in Arenavirus Infection

Among the members of the Arenaviridae family, Lassa virus and Junin virus generate periodic annual outbreaks of severe human hemorrhagic fever (HF) in endemic areas of West Africa and Argentina, respectively. Given the human health threat that arenaviruses represent and the lack of a specific and safe chemotherapy, the search for effective antiviral compounds is a continuous demanding effort. Since diverse host cell pathways and enzymes are used by RNA viruses to fulfill their replicative cycle, the targeting of a host process has turned an attractive antiviral approach in the last years for many unrelated virus types. This strategy has the additional benefit to reduce the serious challenge for therapy of RNA viruses to escape from drug effects through selection of resistant variants triggered by their high mutation rate. This article focuses on novel strategies to identify inhibitors for arenavirus therapy, analyzing the potential for antiviral developments of diverse host factors essential for virus infection.

Single Low-dose Cyclophosphamide Combined with Interleukin-12 Gene Therapy is Superior to a Metronomic Schedule in Inducing Immunity Against Colorectal Carcinoma in Mice

The use of conventional cytotoxic agents at metronomic schedules, alone or in combination with targeted agents or immunotherapy, is being explored as a promising anticancer strategy. We previously reported a potent antitumor effect of a single low-dose cyclophosphamide and interleukin-12 (IL-12) gene therapy against advanced gastrointestinal carcinoma, in mice. Here, we assessed whether the delivery of IL-12 by gene therapy together with metronomic cyclophosphamide exerts antitumor effects in a murine model of colorectal carcinoma. This combination therapy was able, at least in part, to reverse immunosuppression, by decreasing the number of regulatory T cells (Tregs) as well as of splenic myeloid-derived suppressor cells (MDSCs). However, metronomic cyclophosphamide plus IL-12 gene therapy failed to increase the number of tumor-infiltrating T lymphocytes and, more importantly, to induce a specific antitumor immune response. With respect to this, cyclophosphamide at a single low dose displayed a superior anticancer profile than the same drug given at a metronomic schedule. Our results may have important implications in the design of new therapeutic strategies against colorectal carcinoma using cyclophosphamide in combination with immunotherapy.

A Simple Procedure for the Comparison of Covariance Matrices

ABSTRACT: BACKGROUND: Comparing the covariation patterns of populations or species is a basic step in the evolutionary analysis of quantitative traits. Here I propose a new, simple method to make this comparison in two population samples that is based on comparing the variance explained in each sample by the eigenvectors of its own covariance matrix with that explained by the covariance matrix eigenvectors of the other sample. The rationale of this procedure is that the matrix eigenvectors of two similar samples would explain similar amounts of variance in the two samples. I use computer simulation and morphological covariance matrices from the two morphs in a marine snail hybrid zone to show how the proposed procedure can be used to measure the contribution of the matrices orientation and shape to the overall differentiation. RESULTS: I show how this procedure can detect even modest differences between matrices calculated with moderately sized samples, and how it can be used as the basis for more detailed analyses of the nature of these differences. CONCLUSIONS: The new procedure constitutes a useful resource for the comparison of covariance matrices.It could fill the gap between procedures resulting in a single, overall measure of differentiation, and analytical methods based on multiple model comparison not providing such a measure.

RAGE Deficiency Improves Postinjury Sciatic Nerve Regeneration in Type 1 Diabetic Mice

Peripheral neuropathy and insensate limbs and digits cause significant morbidity in diabetic individuals. Previous studies showed that deletion of the receptor for advanced end-glycation products (RAGE) in mice was protective in long-term diabetic neuropathy. Here, we tested the hypothesis that RAGE suppresses effective axonal regeneration in superimposed acute peripheral nerve injury attributable to tissue-damaging inflammatory responses. We report that deletion of RAGE, particularly in diabetic mice, resulted in significantly higher myelinated fiber densities and conduction velocities consequent to acute sciatic nerve crush compared with wild-type control animals. Consistent with key roles for RAGE-dependent inflammation, reconstitution of diabetic wild-type mice with RAGE-null versus wild-type bone marrow resulted in significantly improved axonal regeneration and restoration of function. Diabetic RAGE-null mice displayed higher numbers of invading macrophages in the nerve segments postcrush compared with wild-type animals, and these macrophages in diabetic RAGE-null mice displayed greater M2 polarization. In vitro, treatment of wild-type bone marrow-derived macrophages with advanced glycation end products (AGEs), which accumulate in diabetic nerve tissue, increased M1 and decreased M2 gene expression in a RAGE-dependent manner. Blockade of RAGE may be beneficial in the acute complications of diabetic neuropathy, at least in part, via upregulation of regeneration signals.

Stabilizers of Neuronal and Mitochondrial Calcium Cycling As a Strategy for Developing a Medicine for Alzheimer's Disease

For the last two decades, most efforts on new drug development to treat Alzheimer's disease have been focused to inhibit the synthesis of amyloid beta (Aβ), to prevent Aβ deposition, or to clear up Aβ plaques from the brain of Alzheimer's disease (AD) patients. Other pathogenic mechanisms such as the hyperphosphorylation of the microtubular tau protein (that forms neurofibrillary tangles) have also been addressed as, for instance, with inhibitors of the enzyme glycogen synthase-3 kinase beta (GSK3β). However, in spite of their proven efficacy in animal models of AD, all these compounds have so far failed in clinical trials done in AD patients. It seems therefore desirable to explore new concepts and strategies in the field of drug development for AD. We analyze here our hypothesis that a trifunctional chemical entity acting on the L subtype of voltage-dependent Ca(2+) channels (VDCCs) and on the mitochondrial Na(+)/Ca(2+) exchanger (MNCX), and having additional antioxidant properties, may efficiently delay or stop the death of vulnerable neurons in the brain of AD patients. In recent years, evidence has accumulated indicating that enhanced neuronal Ca(2+) cycling (NCC) and futile mitochondrial Ca(2+) cycling (MCC) are central stage in activating calpain and calcineurin, as well as the intrinsic mitochondrial pathway for apoptosis, leading to death of vulnerable neurons. An additional contributing factor to neuronal death is the excess free radical production linked to distortion of Ca(2+) homeostasis. We propose that an hybrid compound containing a dihydropyridine moiety (to block L channels and mitigate Ca(2+) entry) and a benzothiazepine moiety (to block the MNCX and slow down the rate of Ca(2+) efflux from the mitochondrial matrix into the cytosol), as well as a polyphenol moiety (to sequester excess free radicals) could break down the pathological enhanced NCC and MCC, thus delaying the initiation of apoptosis and the death of vulnerable neurons. In so doing, such a trifunctional compound could eventually become a neuroprotective medicine capable of delaying disease progression in AD patients.

Is the Scale for Measuring Motivational Interviewing Skills a Valid and Reliable Instrument for Measuring the Primary Care Professionals Motivational Skills?: EVEM Study Protocol

ABSTRACT: BACKGROUND: Lifestyle is one of the main determinants of people's health. It is essential to find the most effective prevention strategies to be used to encourage behavioral changes in their patients. Many theories are available that explain change or adherence to specific health behaviors in subjects. In this sense the named Motivational Interviewing has increasingly gained relevance. Few well-validated instruments are available for measuring doctors' communication skills, and more specifically the Motivational Interviewing. METHODS: The hypothesis of this study is that the Scale for Measuring Motivational Interviewing Skills (EVEM questionnaire) is a valid and reliable instrument for measuring the primary care professionals skills to get behavior change in patients. To test the hypothesis we have designed a prospective, observational, multi-center study to validate a measuring instrument. --Scope: Thirty-two primary care centers in Spain. -Sampling and Size: a) face and consensual validity: A group composed of 15 experts in Motivational Interviewing. b) Assessment of the psychometric properties of the scale; 50 physician- patient encounters will be videoed; a total of 162 interviews will be conducted with six standardized patients, and another 200 interviews will be conducted with 50 real patients (n=362). Four physicians will be specially trained to assess 30 interviews randomly selected to test the scale reproducibility. -Measurements for to test the hypothesis: a) Face validity: development of a draft questionnaire based on a theoretical model, by using Delphi-type methodology with experts. b) Scale psychometric properties: intraobservers will evaluate video recorded interviews: content-scalability validity (Exploratory Factor Analysis), internal consistency (Cronbach alpha), intra-/inter-observer reliability (Kappa index, intraclass correlation coefficient, Bland & Altman methodology), generalizability, construct validity and sensitivity to change (Pearson product--moment correlation coefficient). DISCUSSION: The verification of the hypothesis that EVEM is a valid and reliable tool for assessing motivational interviewing would be a major breakthrough in the current theoretical and practical knowledge, as it could be used to assess if the providers put into practice a patient centered communication style and can be used both for training or researching purposes.Trials Registration Dislip-EM studyNCT01282190 (ClinicalTrials.gov).

Marine Products with Anti-Protozoal Activity: A Review

The marine organisms are a rich source of varied natural products with unique functionality. A variety of natural products of new molecular structures with diverse biological activities have been reported from marine flora and fauna for treatment and/or prevention of human diseases. The present review briefly illustrates current status of marine products as antiprotozoal agents. The in vitro and in vivo studies of marine algae, invertebrates and micoorganisms against different protozoa parasites are included. The marine products studied, according international criterions to selection of more promisory products in the differents models reported, demonstrated their potentialities as antiprozoal agents. Herein, the interest of scientific community to search new alternatives from marine environment has been demonstrated.

Relation of Severe Deficiency of Vitamin D to Cardiovascular Mortality During Acute Coronary Syndromes

Vitamin D deficiency is associated with risk for a first cardiovascular event in the general population, possibly because of inflammation, insulin resistance, and neurohumoral activation. However, its relation with outcomes in acute coronary syndromes has not been reported. To test the hypothesis that severe deficiency of vitamin D is independently associated with cardiovascular mortality during ACS, 206 patients admitted for unstable angina, non-ST-segment elevation myocardial infarction, or ST-segment elevation acute myocardial infarction had 25-hydroxyvitamin D serum levels measured at admission. Severe vitamin D deficiency was defined a priori as a value ≤10 ng/ml. The average concentration of vitamin D was 20 ± 8.2 ng/ml, and 10% of patients were severely deficient (95% confidence interval 6.6% to 15%). Cardiovascular mortality during hospitalization took place in 14 patients, an incidence of 6.8%. Patients with severe vitamin D deficiency had in-hospital cardiovascular mortality of 24%, significantly higher than the 4.9% observed in the remaining patients (relative risk 4.3, 95% confidence interval 1.8 to 10, p = 0.001). After adjustment for Global Registry of Acute Coronary Events (GRACE) score, Gensini angiographic score, and potential confounding variables, severe deficiency of vitamin D remained an independent predictor of in-hospital cardiovascular mortality (odds ratio 14, 95% confidence interval 1.2 to 158, p = 0.03). In conclusion, severe vitamin D deficiency is independently associated with in-hospital cardiovascular mortality in patients with acute coronary syndromes.

A Comparative Study of the Efficacy of Intralesional Verapamil Versus Normal Saline Injection in a Novel Peyronie Disease Animal Model: Assessment of Immunohistopathological Changes and Erectile Function Outcome

PURPOSE: While intralesional injections improve penile curvature and decrease plaque volume, the exact mechanism of action on Peyronie disease is unknown. We evaluated penile curvature, immunohistology and erectile function outcomes after intralesional injections of verapamil and normal saline in a previously described Peyronie disease animal model. MATERIALS AND METHODS: Peyronie plaque was induced in 12 adult male rats using an established Peyronie disease animal model. At 4 weeks the rats were divided into group 1-5 with 0.1 mg/0.1 ml intralesional verapamil injected every second day for 2 weeks, group 2-5 with 0.1 ml intralesional normal saline injection and group 3-2 that served as controls. At weeks 6 and 8 penile pressure was measured and serial immunohistochemical staining of penile tissue sections was done. RESULTS: Intralesional injection of verapamil and normal saline resulted in macroscopic and microscopic changes to penile curvature and Peyronie plaque size. Decreased collagen and elastin fibers were measured with a significant reduction in smooth muscle α-actin (p <0.05). Changes were greater in group 1 than group 2 (p <0.05). Intralesional verapamil injection was associated with greater recovery of electrostimulated penile pressure, a surrogate of erectile function, than in the saline and control groups. CONCLUSIONS: To our knowledge this novel study offers for the first time histological evidence of cellular changes and improvement in penile pressure studies in rats with Peyronie plaque after intralesional verapamil injection therapy in a Peyronie disease animal model.

Chemical Characterization, Antioxidant and Cytotoxic Activities of Brazilian Red Propolis

Propolis is known for a long time for its health benefits and biological activities. Here, the red variety from the northeast of Brazil was chemically analyzed and extracts were investigated regarding their antioxidant and antitumor activity. Hydroalcoholic extracts, obtained from the red propolis, revealed polyphenol content, 2,2-diphenyl-1-picrylhydrazyl scavenging potential and enzymatic activities for catalase-like and superoxide dismutase-like. Cytotoxic activity was evaluated for human laryngeal epidermoid carcinoma cell (Hep-2), human cervical adenocarcinoma (HeLa) and human normal epithelial embryonic kidney (Hek-293). Survival analysis for non-tumor cell line showed greater IC50 compared to tumor cell lines, suggesting an increased sensitivity that may correlate with the higher proliferative index of the tumor vs. normal cells. Our results indicate that the Brazilian red propolis is capable of inhibiting cancer cell growth and constitutes an excellent source of antioxidant and antitumor natural agent.

[Deaths by Homicide in Medellin, 1980-2007]

The Colombian city of Medellin has faced a severe problem of violence and homicide. The goal of this retrospective, descriptive-analytical study was to assess the magnitude, distribution, most salient characteristics and possible explanations for the problem between 1980 and 2007. Data were obtained from official databases, hospital records and the available literature, and a focus group was set up. There was a dramatic rise in homicides in the city in the time interval studied, with a total of 84,863 documented murders. Ninety-three percent of the victims were male. The 20 to 29 year-old age group was the most affected. Males in this age group, during the worst year of the period (1991), registered an astounding murder rate of 1,709 per 100,000. Even males in the 15 to 19 year age group and the older brackets saw high murder rates. The most common victims were from the lower and lower-middle socioeconomic strata, as has been observed elsewhere in the region. Serious problems were identified in information and in the provision of services. The impact of homicide on the delivery of healthcare services is discussed and a series of tasks for the healthcare sector in the control of this type of violence is proposed.

Validation in Spanish of a Screening Questionnaire for the Detection of Psoriatic Arthritis in Patients with Psoriasis

Objective. A patient self-administered questionnaire [PsA Screening and Evaluation (PASE)] has been developed and validated in English, but has not been tried in Spanish speaking populations. This study aimed to adapt and validate PASE in Spanish to screen Spanish speaking psoriasis patients for signs and symptoms of inflammatory arthritis.Methods. Initial translation from English to Spanish (forward translation) was performed by two independent translators and the resulting versions were synthesized during a consensus meeting. The questionnaire was tried in a pilot study and resulted in a change in the agreement scale for a frequency scale with wording adaptation [Spanish PASE (PASE-S)].Results. One hundred and eleven patients were screened with PASE-S; 25 with PsA (without previous treatments), 23 with psoriasis, 22 with psoriasis and OA and 41 with OA without psoriasis. The diagnosis of psoriasis was performed by a dermatologist, and a rheumatologist determined the diagnosis of PsA or OA. Patients with PsA had statistically significant higher symptoms, function and total PASE-S scores compared with those without PsA. Receiver operator curves showed an area under the curve of 0.79 (95% CI 0.69, 0.89) for the total score. A cut-off value ≥34 showed sensitivity of 76%, and specificity of 74.4% for the diagnosis of PsA.Conclusion. The validated PASE questionnaire is a self-administered tool that can be used to screen for PsA among patients with psoriasis in a Spanish speaking population. PASE was able to distinguish between symptoms of PsA and OA.

Modulation of Vigilance in the Primary Hypersomnias by Endogenous Enhancement of GABAA Receptors

The biology underlying excessive daytime sleepiness (hypersomnolence) is incompletely understood. After excluding known causes of sleepiness in 32 hypersomnolent patients, we showed that, in the presence of 10 μM γ-aminobutyric acid (GABA), cerebrospinal fluid (CSF) from these subjects stimulated GABA(A) receptor function in vitro by 84.0 ± 40.7% (SD) relative to the 35.8 ± 7.5% (SD) stimulation obtained with CSF from control subjects (Student's t test, t = 6.47, P < 0.0001); CSF alone had no effect on GABA(A) signaling. The bioactive CSF component had a mass of 500 to 3000 daltons and was neutralized by trypsin. Enhancement was greater for α2 subunit- versus α1 subunit-containing GABA(A) receptors and negligible for α4 subunit-containing ones. CSF samples from hypersomnolent patients also modestly enhanced benzodiazepine (BZD)-insensitive GABA(A) receptors and did not competitively displace BZDs from human brain tissue. Flumazenil-a drug that is generally believed to antagonize the sedative-hypnotic actions of BZDs only at the classical BZD-binding domain in GABA(A) receptors and to lack intrinsic activity-nevertheless reversed enhancement of GABA(A) signaling by hypersomnolent CSF in vitro. Furthermore, flumazenil normalized vigilance in seven hypersomnolent patients. We conclude that a naturally occurring substance in CSF augments inhibitory GABA signaling, thus revealing a new pathophysiology associated with excessive daytime sleepiness.

A Cluster-randomised Clinical Trial Comparing Two Cardiovascular Health Education Strategies in a Child Population: the Saving Hearts Project

ABSTRACT: BACKGROUND: This paper describes a methodology for comparing the effects of an eduentertainment strategy involving a music concert, and a participatory class experience involving the description and making of a healthy breakfast, as educational vehicles for delivering obesity-preventing/cardiovascular health messages to children aged 7--8 years.Methods/designThis study will involve a cluster-randomised trial with blinded assessment. The study subjects will be children aged 7--8 years of both sexes attending public primary schools in the Madrid Region. The participating schools (n=30) will be randomly assigned to one of two groups: 1) Group MC, in which the children will attend a music concert that delivers obesity-preventing/cardiovascular health messages, or 2) Group HB, in which the children will attend a participatory class providing the same information but involving the description and making of a healthy breakfast. The main outcome measured will be the increase in the number of correct answers scored on a knowledge questionnaire and in an attitudes test administered before and after the above interventions. The secondary outcome recorded will be the reduction in BMI percentile among children deemed overweight/obese prior to the interventions. The required sample size (number of children) was calculated for a comparison of proportions with an alpha of 0.05 and a beta of 0.20, assuming that the Group MC subjects would show values for the measured variables at least 10% higher than those recorded for the subjects of Group HB. Corrections were made for the design effect and assuming a loss to follow-up of 10%. The maximum sample size required will be 2107 children. Data will be analysed using summary measurements for each cluster, both for making estimates and for hypothesis testing. All analyses will be made on an intention-to-treat basis. DISCUSSION: The intervention providing the best results could be recommended as part of health education for young schoolchildren.Trial registrationClinicaltrials.gov: NCT01418872.

Design and Data Analysis of Experimental Trials to Test Vaccine Candidates Against Zoonotic Pathogens in Animals: the Case of a Clinical Trial Against Campylobacter in Broilers

The development of effective vaccines against zoonotic pathogens represents a priority in public health protection programs. The design of clinical trials and appropriate data analysis of the experiments results are crucial for the assessment of vaccine effectiveness. This manuscript reviews important issues related to the assessment of the effectiveness of vaccines designed to obtain a quantitative reduction of the pathogen in animals or animal products. An effective vaccine will reduce the risk of human infections and therefore the number of human cases. Important considerations will be illustrated using a vaccination trial of a new campylobacter vaccine candidate developed to reduce the numbers of campylobacter in chickens and consequently the numbers of human campylobacteriosis cases. The design of the author's vaccination trial was based on the use of isolators, a parallel group design and several rotations. The effect of clustering or design effect was considered in the sample size calculations. Chickens were randomly assigned to different isolators (treatments) and challenged with Campylobacter jejuni. Samples were obtained at different intervals and processed in the laboratory. C. jejuni counts were determined as colony-forming unit-per-gram of chicken cecum or fecal mass in order to assess vaccine effectiveness. A desired vaccine effect of 2 logs reduction on the numbers of C. jejuni recovered from vaccinated chickens was selected. Sample-size calculations, desired vaccine effect, biological and epidemiological aspects, experimental design and appropriate statistical analysis of data considering group or clustering effects will be the focus of this manuscript.

[Detection of Pulmonary Tumor Thrombosis by Integrated (18F)FDG PET/CT Scans with Intravenous Contrast.]

Use of Speed Profile As Surrogate Measure: Effect of Traffic Calming Devices on Crosstown Road Safety Performance

Urban road safety management is usually characterized by a lack of quantity and quality of crash data and low budgets. However, fifty four percent of road crashes in Spain take place in urban areas. Moreover, ten percent of urban fatal crashes occur on crosstown roads, which are rural roads that traverse small communities. So, traffic calming measures (TCMs) are often implemented in this part of a rural road that traverses a small community in order to reduce both crash frequency and severity by lowering speeds. The objective of the research was to develop a methodology using continuous speed profile to evaluate safety effectiveness of TCMs on crosstown roads. Given the strong relationship between speed and crash experience, safety performance can be related to speed. Consequently, speed can be used indirectly as a surrogate safety measure. Two indexes were defined as surrogate safety measures based on the continuous speed profile: Ra and Ea. Ra represents absolute accumulated speed variations relative to the average speed, and is inversely related to accumulated speed uniformity; and Ea represents accumulated speed variations above the speed limit, and is directly related to accumulated speeding. Naturalistic data was collected using GPS trackers on twelve scenarios with different TCMs spacing. Then, the indexes were applied to individual observed speed profiles (individual analysis) as well as the operating speed profile (global analysis). The values obtained from individual and global analysis were statistically different. Spacing lower than 110m, which was found optimal on previous research, did not allow drivers to modify their speeds as accumulated speed uniformity was quite similar regardless average operating speed; and, accumulated speeding was also minimized. Consequently, scenarios with implemented TCMs according to technical criteria presented a better design quality. On the other hand, age and gender differences did not seem to affect average speed, nor accumulated speed uniformity or accumulated speeding.

Agreement Between Measured and Self-reported Height, Weight and BMI in Predominantly European Middle-aged New Zealanders: Findings from a Nationwide 1989 Survey

To examine the agreement between self-reported and measured height, weight and BMI in 40-50 year old New Zealand men and women.

Multi-locus Phylogenies of the Genus Barteria (Passifloraceae) Portray Complex Patterns in the Evolution of Myrmecophytism

The four species of the central African genus Barteria show variation in habitat and in degree of association with ants. Whereas B. solida, restricted to submontane forests, attracts opportunistic ants to extrafloral nectar, the three other species, found in lowland rainforests (B. fistulosa, B. dewevrei) and in littoral scrub (B. nigritana), possess stem domatia of varying shapes and degrees of specialisation, hosting either non-specific arboreal ants (B. nigritana, some B. dewevrei) or two large species of ants of the genus Tetraponera Smith, 1852 that are specific to some species of Barteria (B. fistulosa, some B. dewevrei). We aimed to investigate whether this variation represents an evolutionary trend toward increasing specialisation of mutualism or the reduction or loss of myrmecophytic traits. For this, we determined phylogenetic relationships within the genus using DNA sequences (primarily nuclear ITS) and microsatellite genotypes (11 loci) on a large sample of individuals, mostly from Cameroon and Gabon. The two types of markers support an initial dichotomy that groups B. dewevrei with B. nigritana and B. fistulosa with B. solida respectively. Within these pairs, species do not appear reciprocally monophyletic. At microsatellite loci, B. nigritana forms a clade embedded within B. dewevrei; and within both B. solida and B. fistulosa, geographical populations show levels of differentiation similar to that observed between populations of B. solida and B. fistulosa. Geographic distance alone does not account for genetic differentiation between species, which indicates reproductive isolation. Divergence in each of the two pairs implies evolutionary transitions in habitat and in myrmecophytism. Specialised mutualism with specific ant species of the genus Tetraponera has been lost in species found in more marginal habitats.

Metallothioneins I/II Are Involved in the Neuroprotective Effect of Sildenafil in Focal Brain Injury

We recently reported that administration of the non-selective GMP-PDE inhibitor zaprinast to cortically cryoinjured rats results three days post-lesion in reduced neuronal cell death that was associated to decreased macrophage/microglial activation and oxidative stress and increased astrogliosis and angiogenesis. Similar effects have been observed in cryoinjured animals overexpressing metallothioneins I/II (MT-I/II), metal-binding cysteine-rich proteins that are up-regulated in response to injury. In this work we have examined the effect of administration of the selective PDE5 inhibitor sildenafil (10mg/kg, sc) 2h before and 24 and 48h after induction of cortical cryolesion in wild type and MT-I/II-deficient mice. Our results show that in wild-type animals sildenafil induces similar changes in glial reactivity, angiogenesis and antioxidant and antiapoptotic effects in the cryolesioned cortex as those observed in rats with zaprinast, indicating that inhibition of PDE5 is responsible for the neuroprotective actions. However, these effects were not observed in mice deficient in MT-I/II. We further show that sildenafil significantly increases MT-I/II protein levels in homogenates of lesioned cortex and MT-I/II immunostaining in glial cells around the lesion. Taken together these results indicate that cGMP-mediated pathways regulate expression of MT-I/II and support the involvement of these proteins in the neuroprotective effects of sildenafil in focal brain lesion.

Effects of the Expression Level of Epidermal Growth Factor Receptor on the Ligand-induced Restructuring of Focal Adhesions: a QCM-D Study

Epidermal growth factor receptor (EGFR) plays a major role in cell migration and invasion and is considered to be the primary source of activation of various malignant tumors. To gain insight into how elevated levels of EGFR influence cellular function, particularly cell motility, we used a quartz crystal microbalance with dissipation monitoring (QCM-D) to examine restructuring of focal adhesions in MCF-10A cells induced by epidermal growth factor. Engineered cells that overexpress epidermal growth factor receptor (EGFR) exhibited a very different kinetic profile from wildtype MCF-10A cells that have a lower level of EGFR with a higher rate for the initial disassembly of focal adhesion and a much lower rate for the later reassembly of focal adhesions. It is conceivable that these effects exhibited by EGFR-overexpressing cells may promote the initiation and maintenance of a more favorable adhesion state for cell migration. This study has demonstrated the capability of the dissipation monitoring function of the QCM-D to quantitatively assess kinetic aspects of cellular processes with a high temporal resolution and sensitivity.

Effects of Habitual Exercise on the EHsp72-Induced Release of Inflammatory Cytokines by Macrophages from Obese Zucker Rats

Regular exercise is a good non-pharmacological treatment of metabolic syndrome in that it improves obesity, diabetes, and inflammation. The 72 kDa extracellular heat shock protein (eHsp72) is released during exercise, thus stimulating the inflammatory responses. The aim of the present work was to evaluate the effect of regular exercise on the eHsp72-induced release of IL-1β, IL-6, and TNFα by macrophages from genetically obese Zucker rats (fa/fa) (ObZ), using lean Zucker (LZ) rats (Fa/fa) to provide reference values. ObZ presented a higher plasma concentration of eHsp72 than LZ, and exercise increased that concentration. In response to eHsp72, the macrophages from ObZ released less IL-1β and TNFα, but more IL-6, than macrophages from LZ. While eHsp72 stimulated the release of IL-1β, TNFα, and IL-6 in the macrophages from healthy LZ (with respect to the constitutive release), it inhibited the release of IL-1β and IL-6 in macrophages from ObZ. The habitual exercise improved the release of inflammatory cytokines by macrophages from ObZ in response to eHsp72 (it increased IL-1β and TNFα, and decreased IL-6), tending to values closer to those determined in healthy LZ. A deregulated macrophage inflammatory and stress response induced by eHsp72 underlies MS, and this is improved by habitual exercise.

High Incidence of Mammary Intraepithelial Neoplasia Development in Men1-disrupted Murine Mammary Glands

Mutations of the MEN1 tumour suppressor gene predispose patients to the development of multiple endocrine neoplasia type 1 (MEN1) syndrome, which is characterised by multiple endocrine tumours, including prolactinomas. The recent findings of the interaction between menin, encoded by the MEN1 gene, and the oestrogen receptor, as well as the observation of rare cases of mammary carcinomas in our heterozygous Men1 mutant mice, led us to investigate a putative tumour suppressor function of the Men1 gene in mouse mammary cells by disrupting the gene in luminal epithelial cells. A significantly higher incidence of mammary intraepithelial neoplasia (MIN) was observed in mutant WapCre-Men1(F/F) mice (51.5%) than in WapCre-Men1(+/+) (0%) or Men1(F/F) (7.1%) control mice. The majority of MIN observed in the mutant mice displayed complete menin inactivation. Because of the leakage of WapCre transgene expression, prolactinomas were observed in 83.3% of mutant mice, leading to premature death. As there was no correlation between MIN development and elevated serum prolactin levels and phospho-STAT5 expression was decreased in mammary lesions, the increased incidence of MIN lesions was most likely due to Men1 disruption rather than to prolactinoma development. Interestingly, in MIN lesions, we found a decrease in membrane-associated E-cadherin and beta-catenin expression, the latter of which is a menin partner. Finally, reduced menin expression was found in a large proportion of two independent cohorts of patients with breast carcinomas. Taken together, the current work indicates a role of Men1 inactivation in the development of mammary pre-cancerous lesions in mice and a potential role in human mammary cancer. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Strategies for Annotation and Curation of Translational Databases: the ETUMOUR Project

The eTUMOUR (eT) multi-centre project gathered in vivo and ex vivo magnetic resonance (MR) data, as well as transcriptomic and clinical information from brain tumour patients, with the purpose of improving the diagnostic and prognostic evaluation of future patients. In order to carry this out, among other work, a database-the eTDB-was developed. In addition to complex permission rules and software and management quality control (QC), it was necessary to develop anonymization, processing and data visualization tools for the data uploaded. It was also necessary to develop sophisticated curation strategies that involved on one hand, dedicated fields for QC-generated meta-data and specialized queries and global permissions for senior curators and on the other, to establish a set of metrics to quantify its contents. The indispensable dataset (ID), completeness and pairedness indices were set. The database contains 1317 cases created as a result of the eT project and 304 from a previous project, INTERPRET. The number of cases fulfilling the ID was 656. Completeness and pairedness were heterogeneous, depending on the data type involved.

The Pediatric Obsessive Compulsive Disorder Treatment Study for Young Children (POTS Jr): Developmental Considerations in the Rationale, Design, and Methods

This paper presents the rationale, design, and methods of the Pediatric Obsessive Compulsive Disorder (OCD) Treatment Study for young children (POTS Jr). The study is a multi-site randomized controlled trial (RCT) of family-based Cognitive Behavioral Treatment (CBT) vs. family-based Relaxation Therapy (RT) for young children (ages 5-8) with OCD, which examines the effect of treatment on symptom reduction, functional impairment, and quality of life. Secondary aims evaluate: potential moderators and mediators of treatment response, differences in time course of response, retention rates, and maintenance of treatment gains over one year post-treatment. The sample included 127 children (53% female) and their parents. With regard to ethnicity, 89% of the sample described themselves as non-Hispanic, 5% Hispanic/Latino, and 6% did not endorse a category. In terms of race, the sample was predominantly (91%) White. Because the rationale and methods of the multi-site RCT have been well established, we emphasize here the methodological aspects of the study that were tailored to meet the developmental needs of young children with OCD. Aspects that are highlighted include: choice of control group, inclusion/exclusion criteria, assessment/measurement issues, treatment adaptations, training, and recruitment.

Novel Phosphorylation Sites in the S. Cerevisiae Cdc13 Protein Reveal New Targets for Telomere Length Regulation

The S. cerevisiae Cdc13 is a multi-functional protein with key roles in regulation of telomerase, telomere end protection, and conventional telomere replication, all of which are cell cycle-regulated processes. Given that phosphorylation is a key mechanism for regulating protein function, we identified sites of phosphorylation using nano liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS). We also determined phosphorylation abundance on both wild type (WT) and a telomerase deficient form of Cdc13, encoded by the cdc13-2 allele, in both G1 phase cells, when telomerase is not active, and G2/M phase cells, when it is. We identified 21 sites of in vivo phosphorylation, of which only five had been reported previously. In contrast, phosphorylation of two in vitro targets of the ATM-like Tel1 kinase, S249 and S255, was not detected. This result helps resolve conflicting data on the importance of phosphorylation of these residues in telomerase recruitment. Multiple residues showed differences in their cell cycle pattern of modification. For example, phosphorylation of S314 was significantly higher in G2/M compared to G1 phase and in WT versus mutant Cdc13, and a S314D mutation negatively affected telomere length. Our findings provide new targets in a key telomerase regulatory protein for modulation of telomere dynamics.

Nucleophosmin/B26 Regulates PTEN Through Interaction with HAUSP in Acute Myeloid Leukemia

PTEN (phosphatase and tensin homolog deleted in chromosome 10) is a bona fide dual lipid and protein phosphatase with cytoplasmic (Cy) and nuclear localization. PTEN nuclear exclusion has been associated with tumorigenesis. Nucleophosmin (NPM1) is frequently mutated in acute myeloid leukemia (AML) and displays Cy localization in mutated nucleophosmin (NPMc+) AML. Here we show that NPM1 directly interacts with herpes virus-associated ubiquitin specific protease (HAUSP), which is known as a PTEN deubiquitinating enzyme. Strikingly, PTEN is aberrantly localized in AML carrying NPMc+. Mechanistically, NPM1 in the nucleus opposes HAUSP-mediated deubiquitination and this promotes the shuttle of PTEN to the cytoplasm. In the cytoplasm, NPMc+ prevents HAUSP from deubiquitinating PTEN, causing the latter to stay in the cytoplasm where it is polyubiquitinated and degraded. Our findings delineate a new NPM1-HAUSP molecular interaction controlling PTEN deubiquitination and trafficking.Leukemia advance online publication, 27 November 2012; doi:10.1038/leu.2012.314.

Toward Greater Inclusion: Lessons from Peru in Confronting Challenges of Multi-sector Collaboration

Despite widespread enthusiasm for broader participation in health policy and programming, little is known about the ways in which multi-sector groups address the challenges that arise in pursuing this goal. Based on the experience of Peru's National Multi-sector Health Coordinating Body (CONAMUSA), this article characterizes these challenges and identifies organizational strategies the group has adopted to overcome them. Comprising nine government ministries, nongovernmental organizations, academia, religious institutions, and international cooperation agencies, CONAMUSA has faced three principal challenges: 1) selecting representatives, 2) balancing membership and leadership across sectors, and 3) negotiating role transition and conflict. In response, the group has instituted a rotation system for formal leadership responsibiliti es, and professionalized management functions; created electoral systems for civil society; and developed conflict of interest guidelines. This case study offers lessons for other countries trying to configure multi-sector groups, and for donors who mandate their creation, tempering unbridled idealism toward inclusive participation with a dose of healthy realism and practical adaptation.

Interactive Applications for Sketch-based Editable Polycube-map

In this paper we propose a sketch-based editable polycube mapping method that, given a general mesh and a simple polycube that coarsely resembles the shape of the object, plus sketched features indicating relevant correspondences between the two, provides a uniform, regular and user-controllable quads-only mesh that can be used as a basis structure for subdivision. Large scale models with complex geometry and topology can be processed efficiently with simple, intuitive operations. We show that the simple, intuitive nature of the polycube map is a substantial advantage from the point of view of the interface by demonstrating a series of applications, including kit-basing, shape morphing, painting over the parameterization domain, and GPU-friendly tessellated subdivision displacement, where the user is also able to control the number of patches in the base mesh by the construction of the base polycube.

Tonoplast Calcium Sensors CBL2 and CBL3 Control Plant Growth and Ion Homeostasis Through Regulating V-ATPase Activity in Arabidopsis

Plant responses to developmental and environmental cues are often mediated by calcium (Ca(2+)) signals that are transmitted by diverse calcium sensors. The calcineurin B-like (CBL) protein family represents calcium sensors that decode calcium signals through specific interactions with a group of CBL-interacting protein kinases. We report functional analysis of Arabidopsis CBL2 and CBL3, two closely related CBL members that are localized to the vacuolar membrane through the N-terminal tonoplast-targeting sequence. While cbl2 or cbl3 single mutant did not show any phenotypic difference from the wild type, the cbl2 cbl3 double mutant was stunted with leaf tip necrosis, underdeveloped roots, shorter siliques and fewer seeds. These defects were reminiscent of those in the vha-a2 vha-a3 double mutant deficient in vacuolar H(+)-ATPase (V-ATPase). Indeed, the V-ATPase activity was reduced in the cbl2 cbl3 double mutant, connecting tonoplast CBL-type calcium sensors to the regulation of V-ATPase. Furthermore, cbl2 cbl3 double mutant was compromised in ionic tolerance and micronutrient accumulation, consistent with the defect in V-ATPase activity that has been shown to function in ion compartmentalization. Our results suggest that calcium sensors CBL2 and CBL3 serve as molecular links between calcium signaling and V-ATPase, a central regulator of intracellular ion homeostasis.Cell Research advance online publication 27 November 2012; doi:10.1038/cr.2012.161.

Epigenetic Regulation in Particulate Matter-Mediated Cardiopulmonary Toxicities: A Systems Biology Perspective

Particulate matter (PM) air pollution exerts significant adverse health effects in global populations, particularly in developing countries with extensive air pollution. Understanding of the mechanisms of PM-induced health effects including the risk for cardiovascular diseases remains limited. In addition to the direct cellular physiological responses such as mitochondrial dysfunction and oxidative stress, PM mediates remarkable dysregulation of gene expression, especially in cardiovascular tissues. The PM-mediated gene dysregulation is likely to be a complex mechanism affected by various genetic and non-genetic factors. Notably, PM is known to alter epigenetic markers (e.g., DNA methylation and histone modifications), which may contribute to air pollution-mediated health consequences including the risk for cardiovascular diseases. Notably, epigenetic changes induced by ambient PM exposure have emerged to play a critical role in gene regulation. Though the underlying mechanism(s) are not completely clear, the available evidence suggests that the modulated activities of DNA methyltransferase (DNMT), histone acetylase (HAT) and histone deacetylase (HDAC) may contribute to the epigenetic changes induced by PM or PM-related chemicals. By employing genome-wide epigenomic and systems biology approaches, PM toxicogenomics could conceivably progress greatly with the potential identification of individual epigenetic loci associated with dysregulated gene expression after PM exposure, as well the interactions between epigenetic pathways and PM. Furthermore, novel therapeutic targets based on epigenetic markers could be identified through future epigenomic studies on PM-mediated cardiopulmonary toxicities. These considerations collectively inform the future population health applications of genomics in developing countries while benefiting global personalized medicine at the same time.

The Genetic Basis of Natural Variation in Oenological Traits in Saccharomyces Cerevisiae

Saccharomyces cerevisiae is the main microorganism responsible for wine alcoholic fermentation. The oenological phenotypes resulting from fermentation, such as the production of acetic acid, glycerol, and residual sugar concentration are regulated by multiple genes and vary quantitatively between different strain backgrounds. With the aim of identifying the quantitative trait loci (QTLs) that regulate oenological phenotypes, we performed linkage analysis using three crosses between highly diverged S. cerevisiae strains. Segregants from each cross were used as starter cultures for 20-day fermentations, in synthetic wine must, to simulate actual winemaking conditions. Linkage analysis on phenotypes of primary industrial importance resulted in the mapping of 18 QTLs. We tested 18 candidate genes, by reciprocal hemizygosity, for their contribution to the observed phenotypic variation, and validated five genes and the chromosome II right subtelomeric region. We observed that genes involved in mitochondrial metabolism, sugar transport, nitrogen metabolism, and the uncharacterized ORF YJR030W explained most of the phenotypic variation in oenological traits. Furthermore, we experimentally validated an exceptionally strong epistatic interaction resulting in high level of succinic acid between the Sake FLX1 allele and the Wine/European MDH2 allele. Overall, our work demonstrates the complex genetic basis underlying wine traits, including natural allelic variation, antagonistic linked QTLs and complex epistatic interactions between alleles from strains with different evolutionary histories.

Maternal Anaemia at Delivery and Haemoglobin Evolution in Children During Their First 18 Months of Life Using Latent Class Analysis

Anaemia during pregnancy and at delivery is an important public health problem in low- and middle-income countries. Its association with the children's haemoglobin level over time remains unclear. Our goals were to identify distinct haemoglobin level trajectories using latent class analysis and to assess the association between these trajectories and maternal anaemia and other risk factors.

Effect of Clonidine in Mice Injected with Tityus Discrepans Scorpion Venom

A study was conducted to assess the effect of clonidine (α(2)-adrenoceptor selective agonist) on glycemia, serum and urine α-amylase, blood urea nitrogen (BUN), serum creatinine, blood leukocyte count, kidney histology and zymogen granule content in pancreatic acini, in mice under the effect of Tityus discrepans (Td) scorpion venom. BALB/c male mice (20 ± 2 g, n=9-15) were injected with a sublethal dose (1 μg/g) of Td venom via intraperitoneal (ip), and were treated (ip) with 0.1 μg/g of clonidine (Catapresan(®)) or saline 30 min after the venom injection, and then every 2 hours. Six hours later, mice were anesthetized with diethylether and urine and blood samples were withdrawn by cystocentesis and cardiocentesis, respectively. Tissue samples were obtained and fixed immediately in buffered formalin (2 %, pH 7.4) and then processed for H&E. Td venom did not cause hyperglycemia, while clonidine induced hyperglycemia and synergized with Td venom. Although the venom did not produce hyperamylasemia, clonidine significantly diminished serum α-amylase activity in envenomed mice. Td venom did not increase significantly urinary α-amylase activity, which was unaffected by clonidine. Morphometric analysis using microphotographs of pancreata from mice injected with Td venom showed a reduced zymogen granule content as judged by the acidophilic bidimensional area of acini. This effect was significantly reduced by clonidine. Kidney samples showed histological changes which were partially affected by the drug. Clonidine reduced the increase in BUN and serum creatinine concentration in envenomed mice. Td venom produced neutrophilia and lymphopenia, which were clonidine-resistant at the assayed dose. These results suggest that α(2)-adrenoceptor selective agonists would be able to reduce some scorpion venom-induced renal and pancreatic disturbances, possibly through the inhibition of neurotransmitter release from presynaptic cholinergic and noradrenergic terminals, as well as from adrenal medulla.

Role of Computed Tomography and Clinical Findings in Pediatric Blunt Intestinal Injury: A Multicenter Study

PURPOSE: Although computed tomographic (CT) scans are accurate in diagnosing solid-organ injuries, their ability to diagnose a blunt intestinal injury (BII) is limited, occasionally requiring repeated imaging. The purpose of this study was to evaluate the role of clinical findings as well as original and repeated CT imaging in the ultimate decision to operate for BII. METHODS: An 18-institution record review of children (≤15 years) diagnosed with a BII confirmed during surgery between 2002 and 2007 was conducted by the American Pediatric Surgery Association Trauma Committee. The incidence of imaging, repeated imaging, and final reported indications for operative exploration were evaluated. RESULTS: Among 331 patients identified with a BII, 292 (88%) underwent at least 1 abdominal CT scan. Sixty-two (19%) underwent at least 1 repeated scan before operation. Forty-seven percent of children who underwent a CT scan were taken to the operating room based primarily on clinical indications (fever, abdominal pain, shock or elevated white blood cell count), whereas 31% were operated on based on both a clinical and CT indication and 22% were operated on based on a CT indication alone (P < 0.001). Although free air was the most common radiographic indication for surgery, 13% of patients with a repeated scan had free air diagnosed on their first CT. Most children undergoing a repeated CT (84%) had findings on the original scan suggesting a BII. Among the 10 patients whose first CT scan result was normal, only 1 went to the operating room based only on radiographic findings. Children who had their first CT scan at a referring hospital were more likely to have a repeated study compared with those imaged at a trauma center (33% vs 13%, P < 0.0001). CONCLUSIONS: Although abdominal CT imaging may contribute to diagnosing intestinal injury after blunt trauma, most children undergo operation based on clinical findings. Repeated imaging should be limited to select patients with diagnostic uncertainty to avoid unneeded delay and radiation exposure.

Activity of Ceftaroline Against Extracellular (broth) and Intracellular (THP-1 Monocytes) Forms of Methicillin-resistant Staphylococcus Aureus: Comparison with Vancomycin, Linezolid and Daptomycin

BACKGROUND: Ceftaroline fosamil is approved for treatment of acute bacterial skin and skin structure infections caused by methicillin-resistant Staphylococcus aureus (MRSA). We examined the activity of its active metabolite (ceftaroline) against intracellular forms of S. aureus in comparison with vancomycin, daptomycin and linezolid. METHODS: Two methicillin-susceptible S. aureus (MSSA) and 11 MRSA strains with ceftaroline MICs from 0.125 to 2 mg/L [two strains vancomycin- and one strain linezolid-resistant (EUCAST interpretative criteria); VISA and cfr+] were investigated. The activity was measured in broth and after phagocytosis by THP-1 monocytes in concentration-dependent experiments (24 h of incubation) to determine: (i) relative potencies (EC(50)) and static concentrations (C(s)) (mg/L and × MIC); and (ii) relative activities at human C(max) (E(C)(max)) and maximal relative efficacies (E(max)) (change in log(10) cfu compared with initial inoculum). Ceftaroline stability and cellular accumulation (at 24 h) were measured by mass spectrometry. RESULTS: Ceftaroline showed similar activities in broth and in monocytes compared with vancomycin, daptomycin and linezolid, with no impact of resistance mechanisms to vancomycin or linezolid. For all four antibiotics, intracellular E(C)(max) and E(max) were considerably lower than in broth (∼0.5 log(10) versus 4-5 log(10) cfu decrease), but the EC(50) and C(s) showed comparatively little change (all values between ∼0.3 and ∼6× MIC). The mean cellular to extracellular ceftaroline concentration ratios (20 mg/L; 24 h) were 0.66 ± 0.05 and 0.90 ± 0.36 in uninfected and infected cells, respectively. CONCLUSION: In vitro, ceftaroline controls the growth of intracellular MRSA to an extent similar to that of vancomycin, linezolid and daptomycin for strains with a ceftaroline MIC ≤2 mg/L.

Multiple-Pass High Pressure Homogenization of Milk for the Development of Pasteurization-like Processing Conditions

Multiple-pass UHPH was used for reducing microbial population of both indigenous spoilage microflora in whole raw milk and a baroresistant pathogen (Staphylococcus aureus) inoculated in whole sterile milk to define pasteurization-like processing conditions. Response surface methodology was followed and multiple response optimization of UHPH operating pressure (OP) (100, 175, 250 MPa) and number of passes (N) (1-5) was conducted through overlaid contour plot analysis. Increasing OP and N had a significant effect (P <0.05) on microbial reduction of both spoilage microflora and S. aureus in milk. Optimized UHPH processes (five 202-MPa passes; four 232-MPa passes) defined a region where a 5- log(10) reduction of total bacterial count of milk and a baro-resistant pathogen are attainable, as a requisite parameter for establishing an alternative method of pasteurization. Multiple-pass UHPH optimized conditions might help producing safe milk without the detrimental effects associated with thermal pasteurization. © 2012 The Authors Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.

Chemoselective Amide Formation Using O-(4-Nitrophenyl)hydroxylamines and Pyruvic Acid Derivatives

A series of O-(4-nitrophenyl)hydroxylamines were synthesized from their respective oximes using a pulsed addition of excess NaBH(3)CN at pH 3 in 65-75% yield. Steric hindrance near the oxime functional group played a key role in both the ease by which the oxime could be reduced and the subsequent reactivity of the respective hydroxylamine. Reaction of the respective hydroxylamines with pyruvic acid derivatives generated the desired amides in good yields. A comparison of phenethylamine systems bearing different leaving groups revealed significant differences in the rates of these systems and suggested that the leaving group ability of the N-OR substituent plays an important role in determining their reactivity with pyruvic acid. Competition experiments (in 68% DMSO/phosphate buffered saline) using 1 equiv of N-phenethyl-O-(4-nitrophenyl)hydroxylamine and 2 equiv of pyruvic acid in the presence of other nucleophiles such as glycine, cysteine, phenol, hexanoic acid, and lysine demonstrated that significant chemoselectivity is present in this reaction. The results suggest that this chemoselective reaction can occur in the presence of excess α-amino acids, phenols, acids, thiols, and amines.

Discovery of Biomarker Combinations That Predict Periodontal Health or Disease with High Accuracy from GCF Samples Based on High-throughput Proteomic Analysis and Mixed-integer Linear Optimization

AIM: To identify optimal combination(s) of proteomic based biomarkers in gingival crevicular fluid (GCF) samples from chronic periodontitis (CP) and periodontally healthy individuals and validate the predictions through known and blind test sets. MATERIALS AND METHODS: GCF samples were collected from 96 CP and periodontally healthy subjects and analysed using high-performance liquid chromatography, tandem mass spectrometry and the PILOT_PROTEIN algorithm. A mixed-integer linear optimization (MILP) model was then developed to identify the optimal combination of biomarkers which could clearly distinguish a blind subject sample as healthy or diseased. RESULTS: A thorough cross-validation of the MILP model capability was performed on a training set of 55 samples and greater than 99% accuracy was consistently achieved when annotating the testing set samples as healthy or diseased. The model was then trained on all 55 samples and tested on two different blind test sets, and using an optimal combination of 7 human proteins and 3 bacterial proteins, the model was able to correctly predict 40 out of 41 healthy and diseased samples. CONCLUSIONS: The proposed large-scale proteomic analysis and MILP model led to the identification of novel combinations of biomarkers for consistent diagnosis of periodontal status with greater than 95% predictive accuracy.

Artery Buckling: New Phenotypes, Models, and Applications

Arteries are under significant mechanical loads from blood pressure, flow, tissue tethering, and body movement. It is critical that arteries remain patent and stable under these loads. This review summarizes the common forms of buckling that occur in blood vessels including cross-sectional collapse, longitudinal twist buckling, and bent buckling. The phenomena, model analyses, experimental measurements, effects on blood flow, and clinical relevance are discussed. It is concluded that mechanical buckling is an important issue for vasculature, in addition to wall stiffness and strength, and requires further studies to address the challenges. Studies of vessel buckling not only enrich vascular biomechanics but also have important clinical applications.

Eating Disorders After Bariatric Surgery: A Case Series

OBJECTIVE: A significant number of post-bariatric surgery (BS) patients present with eating disorders (EDs) symptoms that require specialized treatment. These cases are thought to be underreported due to their frequent sub-syndromal presentation. This article describes ED syndromes that develop subsequent to BS. METHOD: The clinical charts of 12 individuals who were hospitalized on a specialized inpatient EDs unit were reviewed. RESULTS: Based on the new DSM-5 proposed criteria, six patients would meet criteria for an anorexia nervosa (AN) diagnosis: three with binge eating/purge AN subtype and three with restrictive AN subtype. An additional four met criteria for atypical AN, since they were at a normal weight, and two patients met criteria for bulimia nervosa. DISCUSSION: Several similarities to the classical EDs were found. The findings that most distinguished these patients from those with classical EDs were their ages, and the age of onset of the ED for some patients. © 2012 by Wiley Periodicals, Inc. (Int J Eat Disord 2012).

Histomorphometric and Immunohistochemical Study of the Goat Omasum During Prenatal Development

This work studies the morphological changes taking place in the goat omasum during prenatal development, using scanning electron microscope, light microscopy and immunohistochemical analysis. A total of 140 goat embryos and fetuses were used, from the first stages of prenatal life until birth. Differentiation of the omasum as a separate compartment of the primitive gastric tube was observed at 35 days of prenatal life ([crown-rump length (CRL)] 3 cm, 23% gestation). By 38 days (CRL 4.3 cm, 25% gestation) the omasal wall comprised three layers: an internal epithelial layer, a middle layer of pluripotential blastemic tissue and an external layer or serosa. Omasal laminae appeared in the following order: primary at 38 days (CRL 4.3 cm, 25% gestation), secondary at 50 days (CRL 7.7 cm, 33% gestation), tertiary at 59 days (CRL 12 cm, 39% gestation) and quaternary at 64 days (CRL 13.5 cm, 43% gestation). Neuroendocrine cells were detected by synaptophysin (SYP) at 52 days (CRL 8 cm, 35% gestation), while glial cell markers (glial fibrillary acidic protein - GFAP, and vimentin-VIM) were observed at 64 days (CRL 13.5 cm, 43% gestation) and 38 days (CRL 4.3 cm, 25% gestation), respectively. Sympathetic and parasympathetic nerve fibers and nerve bodies were detected via neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) at 95 days (CRL 20 cm, 63% gestation). In conclusion, prenatal development of the omasum - like that of the rumen - appears to take place somewhat earlier in goats than in sheep or cattle, but at a similar stage to that reported in deer.

SPIHT-Based Echocardiogram Compression: Clinical Evaluation and Recommendations of Use

This paper describes an echocardiogram coding method that takes into account the visualization modes in order to compress efficiently the echocardiogram, a methodology to evaluate compressed echocardiograms and the evaluation of the compression method using the proposed evaluation methodology. The compression method takes advantage of the particular characteristics of each visualization mode and uses different compression techniques for each mode to compress efficiently the echocardiogram. A complete evaluation has been designed in order to recommend a minimum transmission rate for each operation mode that guarantees sufficient clinical quality. The evaluation of the echocaradiograms compressed with the proposed method has been carried out. The recommended transmission rates have been established as the following: 200 kbps for the 2D and the color Doppler modes, and 40 kbps for the M and the pulsed/continuous Doppler modes. These rates, especially the latter, are very low compared to previous results. These recommendations are valid for all devices and images compressed with the proposed method. The evaluation process can be applied to any compression method.

Do the Microshear Test Variables Affect the Bond Strength Values?

Little is known about the effect of specimen preparation and testing protocols on the micro-shear bond strength (μSBS) results. To evaluate whether variations in polyethylene rod use affect (μSBS)). Human dentin disks were randomly distributed into six groups (n = 5): polyethylene tube (3 levels) and adhesive system (2 levels). In Group 1, polyethylene tubes filled with polymerized composite) were placed on adhesive covered surfaces. Tubes were removed 24 h after water storage, leaving the rods only. In Group 2, the same procedure was performed; however, tubes were kept in place during testing. In Group 3, composite rods without tubes were placed on adhesive covered dentin. In all groups, adhesives were photoactivated after positioning filled tubes/rods on adhesive covered surfaces. Specimens were tested under shear mode and the data subjected to a two-way ANOVA and Tukey's tests. Groups 1 and 2 resulted in statistically similar mean μSBS (P > 0.05); however, a greater number of pretest failures were observed for Group 1. Higher μSBS values were detected for Group 3, irrespective of adhesive system used (P < 0.05). Removing the polyethylene tube before composite rod is placed on dentin affects μSBS values.

Aporocotyle Mariachristinae N. Sp., and A. Ymakara Villalba & Fernández, 1986 (Digenea: Aporocotylidae) of the Pink Cusk-eel, Genypterus Blacodes (Ophidiiformes: Ophidiidae) from Patagonia, Argentina

Aporocotyle mariachristinae n. sp. and A. ymakara Villalba & Fernández, 1986 were collected from the bulbus arteriosus and ventral aorta of pink cusk-eels, Genypterus blacodes (Forster, 1801) from Patagonia, Argentina. A. mariachristinae n. sp. can be distinguished from all the species of Aporocotyle by the asymmetrical extension of posterior caeca (right posterior caecum longer, terminating at the area between mid-level of ovary and posterior body end; left posterior caecum shorter, terminating at the area between mid-level of cirrus sac and posterior to reproductive organs), the distribution of spines along the ventro-lateral body margins and the number of testes. The new species clearly differs from A. ymakara, from the same host species, in the esophagus / body length ratio, the absence of distal loops at caeca, the anterior caeca / posterior caeca length ratio, and the number of testes. Additionally, in A. ymakara the left posterior caecum may be longer than right posterior caecum, while in the new species left posterior caecum is always shorter. The specimen of A. ymakara collected from Argentina is also described. We also provide observations of the distribution of spines in different species of Aporocotyle, including new specimens of A. argentinensis Smith, 1969 from Merluccius hubbsi Marini, 1933. Molecular sequence data obtained from partial 18S and 28S rDNA regions were compared between the new species and other two species of Aporocotyle (A. argentinensis and A. spinosicanalis Williams, 1958). This is a new locality record for A. ymakara, extending the known geographical distribution for this species from Chile to Argentina, and the first report of two species of Aporocotyle in the same host species and locality.

[Intravenous Drug Preparation Errors in a Neonatal Intensive Care Unit. A Potential Source of Adverse Events.]

INTRODUCTION: To determine the rate of errors during preparation of intravenous drugs in a regional Neonatal Intensive Care Unit (NICU). METHODS: A prospective observational study was performed on 24 non-consecutive working days. The vancomycin and tobramycin solutions administered were analysed to determine drug concentrations. We defined 2 types of error: 1) calculation error, when the deviation between the dose prescribed by the physician and theoretical dose administered, according to calculations performed by the nurse, was greater than ±10%, and 2) precision error, when the deviation between the theoretical concentration and that determined by the laboratory was greater than ±10%. RESULTS: A total of 91 samples were collected, 52 of vancomycin and 39 of tobramycin. Calculation errors were detected in 4.6% of samples. Precision errors were identified in 37.9% of the total sample. CONCLUSIONS: Although the errors reported did not produce adverse clinical consequences, our findings point out a potential source of severe complications. Better methods in the preparation of intravenous medications in NICU are needed.

Interferon Stimulated Genes Are Associated with Peginterferon Plus Ribavirin Treatment Response Regardless IL28B Alleles in HCV/HIV Coinfected Patients

BACKGROUND:: The interaction between interferon stimulated genes (ISGs) expression, IL28B genotypes and hepatitis C treatment outcomes has been mainly evaluated in the liver tissue from HCV-monoinfected patients but with controversial results. Herein, we examined whether more easily accessible peripheral blood mononuclear cells (PBMCs) could be used for this purpose in HIV-HCV-coinfected patients, a population in whom HCV-induced liver disease progression occurs more rapidly and treatment response is lower. METHODS:: Gene expression profiles were examined using the human whole genome Agilent microarray platform in PBMCs collected from HIV/HCV-coinfected patients that had completed a course of peginterferon/ribavirin therapy with validated outcomes. Patients were split out according to the achievement of sustained virological response (SVR) and IL28B rs12979860 genotypes. The GeneSpringGX software was used to select genes differentially expressed in the different groups. RESULTS:: Nineteen HIV/HCV-coinfected individuals receiving antiretroviral therapy and having undetectable plasma HIV-RNA were examined. Global gene expression profiles showed 42 genes differentially expressed according to treatment outcome and 56 according to IL28B genotype. Common genes were not found and functions differed for genes belonging to either group. Whereas at least 26/37 repressed genes (70.3%) in SVR patients were ISGs, none of the 56 differentially expressed genes in carriers of distinct IL28B variants were ISGs (p < 0.0001). CONCLUSIONS:: Baseline expression of ISGs in PBMCs from HCV/HIV coinfected patients influence the response to peginterferon/ribavirin therapy, regardless IL28B genotypes. PBMC specimens can reliably be used for evaluating ISGs expression in clinical practice.

Excess Dietary Methionine Does Not Affect Fracture Healing in Mice

Background: An elevated serum concentration of homocysteine (hyperhomocysteinemia) has been shown to disturb fracture healing. As the essential amino acid, methionine, is a precursor of homocysteine, we aimed to investigate whether excess methionine intake affects bone repair. Material/Methods: We analyzed bone repair in 2 groups of mice. One group was fed a methionine-rich diet (n=13), and the second group received an equicaloric control diet without methionine supplementation (n=12). Using a closed femoral fracture model, bone repair was analyzed by histomorphometry and biomechanical testing at 4 weeks after fracture. Blood was sampled to measure serum concentrations of homocysteine, the bone formation marker osteocalcin, and the bone resorption marker collagen I C-terminal crosslaps Results: Serum concentrations of homocysteine were significantly higher in the methionine group than in the control group, while serum markers of bone turnover did not differ significantly between the 2 groups. Histomorphometry revealed no significant differences in size and tissue composition of the callus between animals fed the methionine-enriched diet and those receiving the control diet. Accordingly, animals of the 2 groups showed a comparable bending stiffness of the healing bones. Conclusions: We conclude that excess methionine intake causes hyperhomocysteinemia, but does not affect fracture healing in mice.

Cancer Care Planning and Quality Improvement: 2 for the Price of 1?

A Simple Method for the Determination of Bioactive Antioxidants in Virgin Olive Oils

BACKGROUND: The importance of olive polyphenols as bioactive compounds has grown in recent years as a result of intensive research on their anticancer, antiatherosclerotic and antihypertensive activities. However, there is currently no official method for determining the content of polyphenols in olive oils because of the technical difficulties in their determination. Here a simple method for the analysis of extra virgin olive oil o-diphenols by visible spectrometry is proposed and compared with the traditional method of solid phase extraction followed by colorimetric determination using sodium molybdate or Folin-Ciocalteu reagent or by high-performance liquid chromatography (HPLC) analysis using UV detection. This new approach to determining total o-diphenolic compounds exploits the oxidation of o-diphenols to quinones in a basic medium. RESULTS: Preliminary results showed a better correlation between the total o-diphenol determination by HPLC and by the proposed method (R(2) = 0.9229) than between the total o-diphenol determination by HPLC and by the molybdate colorimetric method (R(2) = 0.8689). A good correlation was also observed between the total phenolic content determined by HPLC and by the proposed method (R(2) = 0.8196), but this correlation was a little lower than the one obtained between the HPLC method and the Folin-Ciocalteu method (R(2) = 0.8752). CONCLUSION: The proposed method involves very little sample manipulation, requires inexpensive reagents and can be performed in less than 40 min for several samples at the same time, using olive oil samples of only 1-2 g. © 2012 Society of Chemical Industry.

Azo Monomers Exhibiting Low Layer Shrinkage at the SmA-SmC Transition and Trans-cis Light-Induced Isomerization

Methacrylic monomers containing a (phenylene)azobenzene unit substituted with a lateral cyano group and alkyl chains of different length are synthesized and characterized by NMR techniques. Their liquid-crystalline properties are studied by differential scanning calorimetry, polarizing optical microscopy, and X-ray diffraction. All monomers exhibit a mesomorphic behavior that extends over wide temperature ranges with nematic and orthogonal or tilted smectic-type mesophases, depending on the length of the terminal chain. The smectic structures are determined to be single-layered with a low layer shrinkage (<5 %) at the SmA-SmC transition. This atypical behavior is attributed to the combination of a high smectic order promoted by both π-π and bond dipole-bond dipole interactions between cyano-substituted central cores, and a low correlation between neighboring layers arising from dispersive forces between the end groups (methacrylic group and alkyl chain) of the monomer. On the other hand, the trans-cis isomerization of monomers is induced in solution by irradiating with a UV lamp. High cis-isomer contents (≥96 %) are obtained at the photostationary state, which is reached in a relatively short time (40 s).

Growing Latin American Science

Sex Differences in Fear Conditioning in Posttraumatic Stress Disorder

Women are twice as likely as men to develop Posttraumatic Stress Disorder (PTSD). Abnormal acquisition of conditioned fear has been suggested as a mechanism for the development of PTSD. While some studies of healthy humans suggest that women are either no different or express less conditioned fear responses during conditioning relative to men, differences in the acquisition of conditioned fear between men and women diagnosed with PTSD has not been examined.

Preconcentration and Determination of Metal Ions from Fuel Ethanol with a New 2,2'-dipyridylamine Bonded Silica

A silica surface chemically modified with [3-(2,2'-dipyridylamine) propyl] groups was prepared, characterized, and evaluated for its metal ion preconcentration in fuel ethanol. To our knowledge, we are the first authors who have reported the present modification on silica gel surface. The material was characterized using infrared spectra, scanning electronic microscopy, and (13)C and (29)Si solid-state NMR spectra. Batch and column experiments were conducted to investigate for metal ion removal from fuel ethanol. The results showed that the Langmuir model describes the sorption equilibrium data of the metal ions in a satisfactory way. From the Langmuir isotherms, the following maximum adsorption capacities (in mmolg(-1)) were determined: 1.81 for Fe(III), 1.75 for Cr(III), 1.30 for Cu(II), 1.25 for Co(II), 1.15 for Pb(II), 0.95 for Ni(II), and 0.87 for Zn(II). Thermodynamic functions, the change of free energy (ΔG), enthalpy (ΔH), and entropy (ΔS) showed that the adsorption of metal ions onto Si-Pr-DPA was feasible, spontaneous, and endothermic. The sorption-desorption of the metal ions made possible the development of a preconcentration and quantification method of metal ions in fuel ethanol.

Emerging Organic Contaminant Removal Depending on Primary Treatment and Operational Strategy in Horizontal Subsurface Flow Constructed Wetlands: Influence of Redox

This study aimed at assessing the influence of primary treatment (hydrolytic upflow sludge blanket (HUSB) reactor vs. conventional settling) and operational strategy (alternation of saturated/unsaturated phases vs. permanently saturated) on the removal of various emerging organic contaminants (i.e. ibuprofen, diclofenac, acetaminophen, tonalide, oxybenzone, bisphenol A) in horizontal subsurface flow constructed wetlands. For that purpose, a continuous injection experiment was carried out in an experimental treatment plant for 26 days. The plant had 3 treatment lines: a control line (settler-wetland permanently saturated), a batch line (settler-wetland operated with saturate/unsaturated phases) and an anaerobic line (HUSB reactor-wetland permanently saturated). In each line, wetlands had a surface area of 2.95 m(2), a water depth of 25 cm and a granular medium D(60) = 7.3 mm, and were planted with common reed. During the study period the wetlands were operated at a hydraulic and organic load of 25 mm/d and about 4.7 g BOD/m(2)d, respectively. The injection experiment delivered very robust results that show how the occurrence of higher redox potentials within the wetland bed promotes the elimination of conventional quality parameters as well as emerging microcontaminants. Overall, removal efficiencies were always greater for the batch line than for the control and anaerobic lines, and to this respect statistically significantly differences were found for ibuprofen, diclofenac, oxybenzone and bisphenol A. As an example, ibuprofen, whose major removal mechanism has been reported to be biodegradation under aerobic conditions, showed a higher removal in the batch line (85%) than in the control (63%) and anaerobic (52%) lines. Bisphenol A showed also a great dependence on the redox status of the wetlands, finding an 89% removal rate for the batch line, as opposed to the control and anaerobic lines (79 and 65%, respectively). Furthermore, diclofenac showed a greater removal under a higher redox status (70, 48 and 32% in the batch, control and anaerobic lines). Average removal efficiencies of acetaminophen, oxybenzone and tonalide were almost >90% for the 3 treatment lines. The results of this study indicate that the efficiency of horizontal flow constructed wetland systems can be improved by using a batch operation strategy. Furthermore, we tentatively identified 4-hydroxy-diclofenac and carboxy-bisphenol A as intermediate degradation products. The higher abundance of the latter under the batch operation strategy reinforced biodegradation as a relevant bisphenol A removal pathway under higher redox conditions.

Edible and Non-edible Olive Oils Discrimination by the Application of a Sensory Olfactory System Based on Tin Dioxide Sensors

An array of semiconductor sensors has been developed to discriminate virgin olive oil samples based on their organoleptic characteristics. The multisensor, developed at laboratory, is composed by 14 sensing elements of tin dioxide thin layers (doped with Cr and In, and undoped) deposited by the reactive sputtering technique. The sensors are stable and show good repeatability. Off-flavors and extra-virgin olive oil samples, taken at the outlet of the vertical centrifuge of a small experimental olive oil mill and sensory evaluated, have been used. A good discrimination of edible (extra-virgin and virgin) from non-edible (lampante) olive oils has been obtained through the statistical method of principal component analysis (PCA).

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