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Find video protocols related to scientific articles indexed in Pubmed.
Internal structure and standardized scores of the Spanish adaptation of the EGRA (Early Grade Reading Assessment) for early reading assessment.
Psicothema
PUBLISHED: 10-24-2014
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The EGRA (Early Grade Reading Assessment) is an assessment tool containing the main predictors of reading learning disabilities based on the National Reading Panel (NRP) (National Institute of Child Health and Human Development, 2000). This study has two main objectives: First, to analyze the internal structure of the EGRA, and second, to examine the validity and normative data for first and second grade primary school students in a Spanish-Speaking population.
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Certolizumab Pegol, a New Anti-TNF-? in the Armamentarium against Ocular Inflammation.
Ocul. Immunol. Inflamm.
PUBLISHED: 10-18-2014
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Abstract Purpose: To study the efficacy and tolerance of certolizumab pegol (CZP) in active uveitis. Methods: Retrospective case series at 4 referral centers. Patients treated with CZP for active uveitis during at least 6 months were eligible. Inflammation by SUN scores, visual acuity (VA) (logMAR), and central macular thickness (CMT) were compared from baseline until final follow-up. Quiescence was defined as 0+ to 0.5+ in anterior chamber and vitreous haze scores and no CMT increase. Results: Four males and 3 females (14 eyes) were included, mean age 42.4?±?8.8 years. All were long-lasting chronic-relapsing uveitis with prior failure to other anti-TNF-?. After a mean follow-up of 10.4?±?4.8 months, 5/7 patients (71.4%) achieved quiescence with CZP. VA improved significantly from +0.52?±?0.68 to +0.45?±?0.68 (p?=?0.032) at 1 month and to +0.44?±?0.64 (p?=?0.035) at 6 months. No adverse events were found. Conclusion: CZP can be an effective alternative in refractory uveitis.
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Cardiac magnetic resonance imaging findings predict major adverse events in apical hypertrophic cardiomyopathy.
J Thorac Imaging
PUBLISHED: 10-15-2014
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The purpose of this study was to determine the prognostic significance of cardiac magnetic resonance imaging (MRI) findings in patients with apical hypertrophic cardiomyopathy (HCM).
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Fully Porous GaN p-n Junction Diodes Fabricated by Chemical Vapor Deposition.
ACS Appl Mater Interfaces
PUBLISHED: 10-13-2014
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Porous GaN based LEDs produced by corrosion etching techniques demonstrated enhanced light extraction efficiency in the past. However, these fabrication techniques require further postgrown processing steps, which increases the price of the final system. Also, the penetration depth of these etching techniques is limited, and affects not only the semiconductor but also the other elements constituting the LED when applied to the final device. In this paper, we present the fabrication of fully porous GaN p-n junctions directly during growth, using a sequential chemical vapor deposition (CVD) process to produce the different layers that form the p-n junction. We characterized their diode behavior from room temperature to 673 K and demonstrated their ability as current rectifiers, thus proving the potential of these fully porous p-n junctions for diode and LEDs applications. The electrical and luminescence characterization confirm that high electronic quality porous structures can be obtained by this method, and we believe this investigation can be extended to other III-N materials for the development of white light LEDs, or to reduce reflection losses and narrowing the output light cone for improved LED external quantum efficiencies.
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Oxidative stress in Cyprinus carpio induced by hospital wastewater in Mexico.
Ecotoxicology
PUBLISHED: 10-11-2014
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The very wide range of activities performed in hospitals (care, diagnosis, hygiene, maintenance, research) require the use of a large variety of potentially ecotoxic substances such as surfactants, metals, disinfectants and pharmaceuticals. This study aimed to determine oxidative stress in the common carp Cyprinus carpio induced by hospital wastewater (HWW) in Mexico. The median lethal concentration (LC50) and subsequently the lowest observed adverse effect level were determined. Carp were exposed to the latter value (0.5 %) for 24, 48, 72 and 96 h, and the following biomarkers were evaluated in gill, brain, liver and blood: hydroperoxide content (HPC), malondialdehyde (MDA) content, protein carbonyl content (PCC) and activity of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). Significant increases in HPC, MDA content and PCC were observed in exposed specimens, particularly in gill, liver and brain. SOD and CAT activity also increased in liver and brain. In conclusion, this particular HWW induces oxidative stress on C. carpio, this damage being most evident in gill, liver and brain.
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Tau protein provides DNA with thermodynamic and structural features which are similar to those found in histone-DNA complex.
J. Alzheimers Dis.
PUBLISHED: 10-07-2014
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Tau protein has been proposed as a trigger of Alzheimer's disease once it is hyperphosphorylated. However, the role that native tau forms play inside the neuronal nucleus remains unclear. In this work we present results concerning the interaction of tau protein with double-stranded DNA, single-stranded DNA, and also with a histone-DNA complex. The tau-DNA interaction results in a structure resembling the beads-on-a-string form produced by the binding of histone to DNA. DNA retardation assays show that tau and histone induce similar DNA retardation. A surface plasmon resonance study of tau-DNA interaction also confirms the minor groove of DNA as a binding site for tau, similarly to the histone binding. A residual binding of tau to DNA in the presence of Distamycin A, a minor groove binder, suggests the possibility that additional structural domains on DNA may be involved in tau interaction. Finally, DNA melting experiments show that, according to the Zipper model of helix-coil transition, the binding of tau increases the possibility of opening the DNA double helix in isolated points along the chain, upon increasing temperature. This behavior is analogous to histones and supports the previously reported idea that tau could play a protective role in stress situations. Taken together, these results show a similar behavior of tau and histone concerning DNA binding, suggesting that post-translational modifications on tau might impair the role that, by modulating the DNA function, might be attributable to the DNA-tau interaction.
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A long-term macroecological analysis of the recovery of a waterbird metacommunity after site protection.
PLoS ONE
PUBLISHED: 08-18-2014
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We used the so called "land-bridge island" or "nested-subsets" theory to test the resilience of a highly fragmented and perturbated waterbird metacommunity, after legal protection of 18 wetlands in the western Mediterranean. Sites were monitored during 28 years and two seasons per year. The metacommunity was composed by 44 species during breeding and 67 species during wintering, including shorebirds, ducks, herons, gulls and divers (Podicipedidae). We identified a strong nested pattern. Consistent with the fact that the study system was to a large extent a spatial biogeographical continuous for thousands of years, fragmented only during the last centuries due to human activities. Non-random selective extinction was the most likely historical process creating the nested pattern, operated by the differential carrying capacity (surface-area) of the remaining sites. We also found a positive temporal trend in nestedness and a decreasing trend in species turnover among sites (?-diversity), indicating that sites are increasingly more alike to each other (i.e. increased biotic homogenization). This decreasing trend in ?-diversity was explained by an increasing trend in local (?) diversity by range expansion of half the study species. Regional (?) diversity also increased over time, indicating that colonization from outside the study system also occurred. Overall our results suggest that the study metacommunity is recovering from historical anthropogenic perturbations, showing a high long-term resilience, as expected for highly vagile waterbirds. However, not all waterbird groups contributed equally to the recovery, with most breeding shorebird species and most wintering duck species showing no geographical expansion.
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Thermodynamics of the interaction between Alzheimer's disease related tau protein and DNA.
PLoS ONE
PUBLISHED: 08-15-2014
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Tau hyperphosphorylation can be considered as one of the hallmarks of Alzheimer's disease and other tauophaties. Besides its well-known role as a microtubule associated protein, Tau displays a key function as a protector of genomic integrity in stress situations. Phosphorylation has been proven to regulate multiple processes including nuclear translocation of Tau. In this contribution, we are addressing the physicochemical nature of DNA-Tau interaction including the plausible influence of phosphorylation. By means of surface plasmon resonance (SPR) we measured the equilibrium constant and the free energy, enthalpy and entropy changes associated to the Tau-DNA complex formation. Our results show that unphosphorylated Tau binding to DNA is reversible. This fact is in agreement with the protective role attributed to nuclear Tau, which stops binding to DNA once the insult is over. According to our thermodynamic data, oscillations in the concentration of dephosphorylated Tau available to DNA must be the variable determining the extent of Tau binding and DNA protection. In addition, thermodynamics of the interaction suggest that hydrophobicity must represent an important contribution to the stability of the Tau-DNA complex. SPR results together with those from Tau expression in HEK cells show that phosphorylation induces changes in Tau protein which prevent it from binding to DNA. The phosphorylation-dependent regulation of DNA binding is analogous to the Tau-microtubules binding inhibition induced by phosphorylation. Our results suggest that hydrophobicity may control Tau location and DNA interaction and that impairment of this Tau-DNA interaction, due to Tau hyperphosphorylation, could contribute to Alzheimer's pathogenesis.
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Antiplatelet therapy before, during, and after extremity revascularization.
J. Vasc. Surg.
PUBLISHED: 08-08-2014
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Peripheral arterial disease (PAD) is well recognized as a marker for systemic atherosclerosis. Platelets play an essential role in all stages of the disease, contributing to both thrombosis and the development of atherosclerosis. Medication regimens to optimize outcomes in both patients who are to undergo revascularization and those who will be managed without interventional therapy must address antiplatelet therapy. Given the common cardiovascular and cerebrovascular comorbidities in patients with PAD, antiplatelet therapy has the potential to decrease thromboembolic events in addition to improving patency after interventions. This clinical update reviews the current literature and recommendations for antiplatelet therapy in patients with PAD.
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Prevalence of and factors associated with increased arterial stiffness in patients with primary Sjögren's syndrome.
Arthritis Care Res (Hoboken)
PUBLISHED: 08-04-2014
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Objectives: To compare the presence of subclinical atherosclerosis measured by means of pulse wave velocity (PWV) in women with primary Sjögren's syndrome (pSS) versus a healthy age and sex-matched control group, and to identify factors independently associated with PWV in pSS. Patients and methods. Forty four pSS women and 78 age-matched healthy women without overt cardiovascular (CV) diseases were assessed for traditional and non-traditional CV risk factors. PWV was also performed. A linear regression analysis was used to identify factors independently associated with PWV in pSS. Results. Women with pSS had significantly higher PWV than controls (p=0.030) and the frequency of increased PWV was significantly higher in this group (25% vs. 8%; p=0.013). The proportion of patients aged ?50 years (ratio 4.6) with increased PWV was almost 2-fold higher than those aged >50 years (ratio 2.4) with respect to controls. Positivity for anti-SSB was more frequent in patients with normal PWV than in those with increased PWV (61% vs. 18%; p=0.034). Women with pSS and increased PWV had lower levels of 25(OH) vitamin D (25(OH)D) (p=0.047) than pSS patients with normal PWV. In addition, 25(OH)D levels tended to correlate inversely with PWV in pSS women (p=0.067) but not in controls (p=0.97). In the multivariate analysis FRS and the Sjögren's syndrome damage index (SSDI) emerged as factors independently correlated with PWV. Conclusions. Women with pSS had higher PWV than controls but similar FRS. FRS and chronic damage were found to be independently associated with PWV. © 2014 American College of Rheumatology.
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[Effect of age on the prevalence of diabetes mellitus in Spain between 2001 and 2012].
Nutr Hosp
PUBLISHED: 06-29-2014
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To assess the effect of age on the increase in DM prevalence in Spain between 2001 and 2012.
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Multiple length peptide-pheromone variants produced by Streptococcus pyogenes directly bind Rgg proteins to confer transcriptional regulation.
J. Biol. Chem.
PUBLISHED: 06-23-2014
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Streptococcus pyogenes, a human-restricted pathogen, accounts for substantial mortality related to infections worldwide. Recent studies indicate that streptococci produce and respond to several secreted peptide signaling molecules (pheromones), including those known as short hydrophobic peptides (SHPs), to regulate gene expression by a quorum-sensing mechanism. Upon transport into the bacterial cell, pheromones bind to and modulate activity of receptor proteins belonging to the Rgg family of transcription factors. Previously, we reported biofilm regulation by the Rgg2/3 quorum-sensing circuit in S. pyogenes. The aim of this study was to identify the composition of mature pheromones from cell-free culture supernatants that facilitate biofilm formation. Bioluminescent reporters were employed to detect active pheromones in culture supernatants fractionated by reverse-phase chromatography, and mass spectrometry was used to characterize their properties. Surprisingly, multiple SHPs that varied by length were detected. Synthetic peptides of each variant were tested individually using bioluminescence reporters and biofilm growth assays, and although activities differed widely among the group, peptides comprising the C-terminal eight amino acids of the full-length native peptide were most active. Direct Rgg/SHP interactions were determined using a fluorescence polarization assay that utilized FITC-labeled peptide ligands. Peptide receptor affinities were seen to be as low as 500 nm and their binding affinities directly correlated with observed bioactivity. Revelation of naturally produced pheromones along with determination of their affinity for cognate receptors are important steps forward in designing compounds whose purpose is positioned for future therapeutics aimed at treating infections through the interference of bacterial communication.
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Allogeneic peripheral blood stem cell transplantation using reduced-intensity conditioning in an outpatient setting in ABO-incompatible patients: are survival and graft-versus-host disease different?
Transfusion
PUBLISHED: 06-06-2014
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Graft-versus-host disease (GVHD) is a major cause of morbimortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Minor ABO incompatibility has been associated with an increased risk of GVHD. We analyzed the impact of ABO matching on patient outcome after peripheral blood, reduced-intensity allo-HSCT in an outpatient setting, and its relationship with GVHD.
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Genetic and biochemical evidence that recombinant Enterococcus spp. strains expressing gelatinase (GelE) produce bovine milk-derived hydrolysates with high angiotensin converting enzyme-inhibitory activity (ACE-IA).
J. Agric. Food Chem.
PUBLISHED: 06-06-2014
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In this work, genes encoding gelatinase (gelE) and serine proteinase (sprE), two extracellular proteases produced by Enterococcus faecalis DBH18, were cloned in the protein expression vector pMG36c, containing the constitutive P32 promoter, generating the recombinant plasmids pCG, pCSP, and pCGSP encoding gelE, sprE, and gelE-sprE, respectively. Transformation of noncaseinolytic E. faecalis P36, E. faecalis JH2-2, E. faecium AR24, and E. hirae AR14 strains with these plasmids permitted detection of caseinolytic activity only in the strains transformed with pCG or pCGSP. Complementation of a deletion (knockout) mutant of E. faecalis V583 for production of gelatinase (GelE) with pCG unequivocally supported that gelE is responsible for the caseinolytic activity of the transformed strain grown in bovine skim milk (BSM). RP-HPLC-MS/MS analysis of hydrolysates of transformed Enterococcus spp. strains grown in BSM permitted the identification of 38 major peptide fragments including peptides with previously reported angiotensin converting enzyme-inhibitory activity (ACE-IA), antihypertensive activity, and antioxidant activity.
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[Mobility, traffic accidents, and associated factors among Guatemalan university students].
Cad Saude Publica
PUBLISHED: 06-05-2014
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The aim of this study was to identify and quantify the association between the amount of driving (km/year), traffic accidents, and other factors among university students in Guatemala. A cross-sectional study was performed during the 2010-2011 school year in a sample of 1,016 drivers who completed a self-administered questionnaire that assessed mobility patterns, use of safety accessories, driving style, and automobile crashes. The results showed a positive association between amount of driving and greater involvement in risky driving (adjusted regression coefficient 3.25, 95%CI: 2.23-4.27, for the highest level of exposure). More frequent involvement in risky driving and older age showed the strongest associations with traffic accidents. Although the amount of driving was positively associated with a higher accident rate, most of this association was found to be mediated by involvement in risky driving practices.
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Metals and nonsteroidal anti-inflammatory pharmaceuticals drugs present in water from Madín Reservoir (Mexico) induce oxidative stress in gill, blood, and muscle of common carp (Cyprinus carpio).
Arch. Environ. Contam. Toxicol.
PUBLISHED: 05-09-2014
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Many toxic xenobiotics that enter the aquatic environment exert their effects through redox cycling. Oxidative stress, which incorporates both oxidative damage and antioxidant defenses, is a common effect induced in organisms exposed to xenobiotics in their environment. The results of the present study aimed to determine the oxidative stress induced in the common carp Cyprinus carpio by contaminants [metals and nonsteroidal anti-inflammatory drugs (NSAIDs)] present in Madín Reservoir. Five sampling stations (SSs), considered to have the most problems due to discharges, were selected. Carp were exposed to water from each SS for 96 h, and the following biomarkers were evaluated in gill, blood, and muscle: hydroperoxide content, lipid peroxidation, protein carbonyl content, and the activity of antioxidant enzymes superoxide dismutase and catalase. Results show that contaminants (metals and NSAIDs) present in water from the different SSs induce oxidative stress. Thus, water in this reservoir is contaminated with xenobiotics that are hazardous to C. carpio, a species consumed by the local human population.
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Contemplating the ultimate sacrifice: identity fusion channels pro-group affect, cognition, and moral decision making.
J Pers Soc Psychol
PUBLISHED: 04-23-2014
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Although most people acknowledge the moral virtue in sacrificing oneself to save others, few actually endorse self-sacrifice. Seven experiments explored the cognitive and emotional mechanisms that underlie such endorsements. Participants responded to 1 of 2 moral dilemmas in which they could save 5 members of their country only by sacrificing themselves. Over 90% of participants acknowledged that the moral course of action was to sacrifice oneself to save others (Experiment 1), yet only those who were strongly fused with the group preferentially endorsed self-sacrifice (Experiments 2-7). The presence of a concern with saving group members rather than the absence of a concern with self-preservation motivated strongly fused participants to endorse sacrificing themselves for the group (Experiment 3). Analyses of think aloud protocols suggested that saving others was motivated by emotional engagement with the group among strongly fused participants but by utilitarian concerns among weakly fused participants (Experiment 4). Hurrying participants' responses increased self-sacrifice among strongly fused participants but decreased self-sacrifice among weakly fused participants (Experiment 5). Priming the personal self increased endorsement of self-sacrifice among strongly fused participants but further reduced endorsement of self-sacrifice among weakly fused participants (Experiment 6). Strongly fused participants ignored utilitarian considerations, but weakly fused persons endorsed self-sacrifice more when it would save more people (Experiment 7). Apparently, the emotional engagement with the group experienced by strongly fused persons overrides the desire for self-preservation and compels them to translate their moral beliefs into self-sacrificial behavior.
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Antibody and cytokine responses to Giardia excretory/secretory proteins in Giardia intestinalis-infected BALB/c mice.
Parasitol. Res.
PUBLISHED: 04-16-2014
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The humoral and cellular responses against excretory/secretory proteins and soluble extracts of Giardia intestinalis were evaluated in the course of experimental G. intestinalis infection in BALB/c mice. Production of IgG1, IgG2a, IgA, and IgE antibodies against excreted/secreted proteins and soluble extract was detected after infection by G. intestinalis. Specific IgA antibody against E/S proteins and soluble extract form intestinal fluids in infected mice was detected by ELISA. The Western blotting identified proteins of 30, 58, 63, and 83 kDa for IgA and IgG, respectively. High proliferation rate in vitro of spleen cell and secretion of interleukin-4 (IL-4) at 21 days p.i. after stimulation with excreted/secreted proteins and low proliferative response in the presence of soluble extract in infected BALB/c mice was observed. High production of interferon gamma (IFN-?) and interleukin-5 (IL-5) at the time of decreasing cyst output (14-21 days p.i.) in infected mice was recorded, suggesting the important role of these cytokines in the control of the infection. Interestingly, progressive and gradual increase of the interleukin-10 after stimulation with both preparations was recorded from 7 days until 28 days after infection, indicating the possible regulatory effect of these antigens on the immune response during Giardia infection. Therefore, the infection by Giardia duodenalis stimulates a mixed response Th1 and Th2, mainly stimulated by excretory/secretory antigens. The immunogenicity of these antigens may be a suitable for identification of the proteins related with the effective immune response in the course of infection by G. duodenalsis.
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Relationship between homocysteine levels and hypertension in systemic lupus erythematosus.
Arthritis Care Res (Hoboken)
PUBLISHED: 03-25-2014
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Homocysteine has been linked to atherosclerosis and hypertension (HT) in the general population. However, there is limited evidence regarding the effect of homocysteine on blood pressure and arterial stiffness in systemic lupus erythematosus (SLE). We examined whether homocysteine is associated with HT and arterial stiffness in women with SLE.
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Non-weight-bearing status compromises the functional level up to 1 yr after hip fracture surgery.
Am J Phys Med Rehabil
PUBLISHED: 03-25-2014
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The aim of this study was to examine the influence of weight-bearing (WB) status after hip fracture surgery on 1-yr functional outcome.
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Computed tomography and magnetic resonance imaging of the coronary sinus: anatomic variants and congenital anomalies.
Insights Imaging
PUBLISHED: 03-10-2014
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The coronary sinus (CS) is an important vascular structure that allows for access into the coronary veins in multiple interventional cardiology procedures, including catheter ablation of arrhythmias, pacemaker implantation and retrograde cardioplegia. The success of these procedures is facilitated by the knowledge of the CS anatomy, in particular the recognition of its variants and anomalies. This pictorial essay reviews the spectrum of CS anomalies, with particular attention to the distinction between clinically benign variants and life-threatening defects. Emphasis will be placed on the important role of cardiac CT and cardiovascular magnetic resonance in providing detailed anatomic and functional information of the CS and its relationship to surrounding cardiac structures. Teaching Points • Cardiac CT and cardiovascular magnetic resonance offer 3D high-resolution mapping of the coronary sinus in pre-surgical planning.• Congenital coronary sinus enlargement occurs in the presence or absence of a left-to-right shunt.• Lack of recognition of coronary sinus anomalies can lead to adverse outcomes in cardiac procedures.• In coronary sinus ostial atresia, coronary venous drainage to the atria occurs via Thebesian or septal veins.• Coronary sinus diverticulum is a congenital outpouching of the coronary sinus and may predispose to cardiac arrhythmias.
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Chronic oral or intraarticular administration of docosahexaenoic acid reduces nociception and knee edema and improves functional outcomes in a mouse model of Complete Freund's Adjuvant-induced knee arthritis.
Arthritis Res. Ther.
PUBLISHED: 02-20-2014
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Clinical and preclinical studies have shown that supplementation with ?-3 polyunsaturated fatty acids (?-3 PUFAs) reduce joint destruction and inflammation present in rheumatoid arthritis (RA). However, the effects of individual ?-3 PUFAs on chronic arthritic pain have not been evaluated to date. Thus, our aim in this study was to examine whether purified docosahexaenoic acid (DHA, an ?-3 PUFA) reduces spontaneous pain-related behavior and knee edema and improves functional outcomes in a mouse model of knee arthritis.
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Oleic acid modulates mRNA expression of liver X receptor (LXR) and its target genes ABCA1 and SREBP1c in human neutrophils.
Eur J Nutr
PUBLISHED: 02-19-2014
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Regulation of liver X receptors (LXRs) is essential for cholesterol homeostasis and inflammation. The present study was conducted to determine whether oleic acid (OA) could regulate mRNA expression of LXR? and LXR?-regulated genes and to assess the potential promotion of oxidative stress by OA in neutrophils.
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Proteome-wide search for PP2A substrates in fission yeast.
Proteomics
PUBLISHED: 02-19-2014
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PP2A (protein phosphatase 2A) is a major phosphatase in eukaryotic cells that plays an essential role in many processes. PP2A mutations in Schizosaccharomyces pombe result in defects of cell cycle control, cytokinesis and morphogenesis. Which PP2A substrates are responsible for these changes is not known. In this work, we searched for PP2A substrates in S. pombe using two approaches, 2D-DIGE analysis of PP2A complex mutants and identification of PP2A interacting proteins. In both cases, we used MS to identify proteins of interest. In the DIGE experiment, we compared proteomes of wild-type S. pombe, deletion of pta2, the phosphoactivator of the PP2A catalytic subunit, and pab1-4, a mutant of B-type PP2A regulatory subunit. A total of 1742 protein spots were reproducibly resolved by 2D-DIGE and 51 spots demonstrated significant changes between PP2A mutants and the wild-type control. MS analysis of these spots identified 27 proteins that include key regulators of glycerol synthesis, carbon metabolism, amino acid biosyntesis, vitamin production, and protein folding. Importantly, we independently identified a subset of these proteins as PP2A binding partners by affinity precipitation, suggesting they may be direct targets of PP2A. We have validated our approach by demonstrating that phosphorylation of Gpd1, a key enzyme in glycerol biogenesis, is regulated by PP2A and that ability of cells to respond to osmotic stress by synthesizing glycerol is compromised in the PP2A mutants. Our work contributes to a better understanding of PP2A function and identifies potential PP2A substrates.
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Effectiveness of an occupational therapy intervention in reducing emotional distress in informal caregivers of hip fracture patients: a randomized controlled trial.
Clin Rehabil
PUBLISHED: 02-17-2014
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To examine the effectiveness of an occupational therapy intervention program in reducing emotional distress in informal caregivers of hip fracture patients.
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Use of synthetic genes for cloning, production and functional expression of the bacteriocins enterocin A and bacteriocin E 50-52 by Pichia pastoris and Kluyveromyces lactis.
Mol. Biotechnol.
PUBLISHED: 02-11-2014
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The use of synthetic genes may constitute a successful approach for the heterologous production and functional expression of bacterial antimicrobial peptides (bacteriocins) by recombinant yeasts. In this work, synthetic genes with adapted codon usage designed from the mature amino acid sequence of the bacteriocin enterocin A (EntA), produced by Enterococcus faecium T136, and the mature bacteriocin E 50-52 (BacE50-52), produced by E. faecium NRRL B-32746, were synthesized. The synthetic entA and bacE50-52 were cloned into the protein expression vectors pPICZ?A and pKLAC2 for transformation of derived vectors into Pichia pastoris X-33 and Kluyveromyces lactis GG799, respectively. The recombinant vectors were linearized and transformed into competent cells selecting for P. pastoris X-33EAS (entA), P. pastoris X-33BE50-52S (bacE50-52), K. lactis GG799EAS (entA), and K. lactis GG799BE50-52S (bacE50-52). P. pastoris X-33EAS and K. lactis GG799EAS, but not P. pastoris X-33BE50-52S and K. lactis GG799BE50-52S, showed antimicrobial activity in their supernatants. However, purification of the supernatants of the producer yeasts permitted recovery of the bacteriocins EntA and BacE50-52. Both purified bacteriocins were active against Gram-positive bacteria such as Listeria monocytogenes but not against Gram-negative bacteria, including Campylobacter jejuni.
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Syntactic Awareness and Arithmetic Word Problem Solving in Children With and Without Learning Disabilities.
J Learn Disabil
PUBLISHED: 02-11-2014
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Arithmetic word problem (AWP) solving is a highly demanding task for children with learning disabilities (LD) since verbal and mathematical information have to be integrated. This study examines specifically how syntactic awareness (SA), the ability to manage the grammatical structures of language, affects AWP solving. Three groups of children in elementary education were formed: children with arithmetic learning disabilities (ALD), children with reading learning disabilities (RLD), and children with comorbid arithmetic and reading learning disabilities (ARLD). Mediation analysis confirmed that SA was a mediator variable for both groups of children with reading disabilities when solving AWPs, but not for children in the ALD group. All groups performed below the control group in the problem solving task. When SA was controlled for, semantic structure and position of the unknown set were variables that affected both groups with ALD. Specifically, children with ALD only were more affected by the place of the unknown set.
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Diagnosis of latent tuberculosis in patients with systemic lupus erythematosus: T.SPOT.TB versus tuberculin skin test.
Biomed Res Int
PUBLISHED: 02-07-2014
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Early studies in patients with systemic lupus erythematosus (SLE) reported increased incidence of tuberculosis. The tuberculin skin test (TST) is the technique of choice to detect latent tuberculosis infection (LTBI) but has several limitations.
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Determination of calcium disodium ethylenediaminetetraacetate (E385) in marketed bottled legumes, artichokes and emulsified sauces by gas chromatography with mass spectrometric detection.
Food Chem
PUBLISHED: 01-22-2014
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An original method to determine the food additive calcium disodium ethylenediaminetetraacetate in bottled food is proposed. The method involves the solid-liquid extraction of a portion of the whole content of legume or artichoke bottles, or the dilution of sauce samples, with water followed by an evaporation step by heating. Finally, ethylenediaminetetraacetic acid is methylated and determined by GC. Recoveries obtained on spiked samples were acceptable (96-108%) with RSDs comprised from 4.3% to 10%. Results suggest that the determination of additive only in the liquid phase of legume or artichoke bottles is not suitable to know its total amount because the additive is distributed between the liquid and solid phases. The contribution of each step of the analytical method to the uncertainty of the measured concentration has been assessed by a "bottom-up" approach, including the heterogeneity of the sample which resulted to be very variable after studying twenty samples.
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A zero-fluoroscopy approach to cavotricuspid isthmus catheter ablation: comparative analysis of two electroanatomical mapping systems.
Pacing Clin Electrophysiol
PUBLISHED: 01-21-2014
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Electroanatomical mapping systems have reduced the amount of fluoroscopy required to ablate the cavotricuspid isthmus. The aims of this study are to evaluate the feasibility and safety of a zero-fluoroscopy approach to cavotricuspid isthmus catheter ablation using the Carto®3 system (Biosense Webster, Diamond Bar, CA, USA) and to compare the results of this approach with those of the zero-fluoroscopy approach using the Ensite-NavX™ system (St. Jude Medical, St. Paul, MN, USA).
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[Immunomodulatory properties of stem mesenchymal cells in autoimmune diseases.]
Med Clin (Barc)
PUBLISHED: 01-09-2014
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Autoimmune diseases are a cluster of disorders characterized by a failure of the immune tolerance and a hyperactivation of the immune system that leads to a chronic inflammation state and the damage of several organs. The medications currently used to treat these diseases usually consist of immunosuppressive drugs that have significant systemic toxic effects and are associated with an increased risk of opportunistic infections. Recently, several studies have demonstrated that mesenchymal stem cells have immunomodulatory properties, a feature that make them candidates to be used in the treatment of autoimmune diseases. In the present study, we reviewed the role of this therapy in the treatment of systemic lupus erythematosus, Sjögren's syndrome, systemic sclerosis, Crohn's disease and multiple sclerosis, as well as the potential risks associated with its use.
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uORF, a regulatory mechanism of the Arabidopsis polyamine oxidase 2.
Mol. Biol. Rep.
PUBLISHED: 01-04-2014
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The translational efficiency of an mRNA can be modulated by elements located in the 5'-untranslated region. The flavin-containing polyamine oxidases catabolize oxidative deamination of spermidine and spermine, thus contributing to polyamine homeostasis as well as diverse biological processes through their reaction products. In this study, we characterized the uORF of AtPAO2 gene using the GUS reporter gene. Transgenic lines harboring the native AtPAO2 promoter or the constitutive CaMV 35S promoter show that the uORF negatively affects GUS expression. Exogenous applications of PAs positively modulate GUS expression, thus alleviating the negative effect of AtPAO2 uORF, while treatments with MGBG inhibitor show an opposite effect. Our data suggest that AtPAO2 uORF regulatory mechanism is modulated by polyamines. In addition, we present a comparative in silico study of the uORFs identified in several plant transcripts encoding polyamine oxidases in both mono- and dicotyledonous plants as well as in the Bryophyte Physcomitrella patens. The polyamine oxidase uORF-encoded peptides are conserved among families and share conserved features such as their position, length, and amino acid sequence. Our findings provide new insights into the regulatory mechanism of polyamine oxidase genes and encourage further exploration to assess the biological significance of uORFs in the polyamine catabolic pathway.
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A dehydrin-dehydrin interaction: the case of SK3 from Opuntia streptacantha.
Front Plant Sci
PUBLISHED: 01-01-2014
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Dehydrins belongs to a large group of highly hydrophilic proteins known as Late Embryogenesis Abundant (LEA) proteins. It is well known that dehydrins are intrinsically disordered plant proteins that accumulate during the late stages of embryogenesis and in response to abiotic stresses; however, the molecular mechanisms by which their functions are carried out are still unclear. We have previously reported that transgenic Arabidopsis plants overexpressing an Opuntia streptacantha SK3 dehydrin (OpsDHN1) show enhanced tolerance to freezing stress. Herein, we show using a split-ubiquitin yeast two-hybrid system that OpsDHN1 dimerizes. We found that the deletion of regions containing K-segments and the histidine-rich region in the OpsDHN1 protein affects dimer formation. Not surprisingly, in silico protein sequence analysis suggests that OpsDHN1 is an intrinsically disordered protein, an observation that was confirmed by circular dichroism and gel filtration of the recombinantly expressed protein. The addition of zinc triggered the association of recombinantly expressed OpsDHN1 protein, likely through its histidine-rich motif. These data brings new insights about the molecular mechanism of the OpsDHN1 SK3-dehydrin.
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Quorum sensing in group A Streptococcus.
Front Cell Infect Microbiol
PUBLISHED: 01-01-2014
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Quorum sensing (QS) is a widespread phenomenon in the microbial world that has important implications in the coordination of population-wide responses in several bacterial pathogens. In Group A Streptococcus (GAS), many questions surrounding QS systems remain to be solved pertaining to their function and their contribution to the GAS lifestyle in the host. The QS systems of GAS described to date can be categorized into four groups: regulator gene of glucosyltransferase (Rgg), Sil, lantibiotic systems, and LuxS/AI-2. The Rgg family of proteins, a conserved group of transcription factors that modify their activity in response to signaling peptides, has been shown to regulate genes involved in virulence, biofilm formation and competence. The sil locus, whose expression is regulated by the activity of signaling peptides and a putative two-component system (TCS), has been implicated on regulating genes involved with invasive disease in GAS isolates. Lantibiotic regulatory systems are involved in the production of bacteriocins and their autoregulation, and some of these genes have been shown to target both bacterial organisms as well as processes of survival inside the infected host. Finally AI-2 (dihydroxy pentanedione, DPD), synthesized by the LuxS enzyme in several bacteria including GAS, has been proposed to be a universal bacterial communication molecule. In this review we discuss the mechanisms of these four systems, the putative functions of their targets, and pose critical questions for future studies.
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Physiological and molecular implications of plant polyamine metabolism during biotic interactions.
Front Plant Sci
PUBLISHED: 01-01-2014
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During ontogeny, plants interact with a wide variety of microorganisms. The association with mutualistic microbes results in benefits for the plant. By contrast, pathogens may cause a remarkable impairment of plant growth and development. Both types of plant-microbe interactions provoke notable changes in the polyamine (PA) metabolism of the host and/or the microbe, being each interaction a complex and dynamic process. It has been well documented that the levels of free and conjugated PAs undergo profound changes in plant tissues during the interaction with microorganisms. In general, this is correlated with a precise and coordinated regulation of PA biosynthetic and catabolic enzymes. Interestingly, some evidence suggests that the relative importance of these metabolic pathways may depend on the nature of the microorganism, a concept that stems from the fact that these amines mediate the activation of plant defense mechanisms. This effect is mediated mostly through PA oxidation, even though part of the response is activated by non-oxidized PAs. In the last years, a great deal of effort has been devoted to profile plant gene expression following microorganism recognition. In addition, the phenotypes of transgenic and mutant plants in PA metabolism genes have been assessed. In this review, we integrate the current knowledge on this field and analyze the possible roles of these amines during the interaction of plants with microbes.
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Feedback Regulation of SIN by Etd1 and Rho1 in Fission Yeast.
Genetics
PUBLISHED: 12-13-2013
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In fission yeast, the septation initiation network (SIN) is thought to promote cytokinesis by downstream activation of Rho1, a conserved GTPase that controls cell growth and division. Here we show that Etd1 and PP2A-Pab1, antagonistic regulators of SIN, are Rho1 regulators. Our genetic and biochemical studies indicate that a C-terminal region of Etd1 may activate Rho1 by directly binding it, whereas an N-terminal domain confers its ability to localize at the growing tips and the division site where Rho1 functions. In opposition to Etd1, our results indicate that PP2A-Pab1 inhibits Rho1. The SIN cascade is upstream regulated by the Spg1 GTPase. In the absence of Etd1, activity of Spg1 drops down prematurely, thereby inactivating SIN. Interestingly, we find that ectopic activation of Rho1 restores Spg1 activity in Etd1-depleted cells. By using a cytokinesis block strategy, we show that Rho1 is essential to feedback activate Spg1 during actomyosin ring constriction. Therefore, activation of Spg1 by Rho1, which in turn is regulated by Etd1, uncovers a novel feedback loop mechanism that ensures SIN activity while cytokinesis is progressing.
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Effect of occupational therapy on functional and emotional outcomes after hip fracture treatment: a randomized controlled trial.
Clin Rehabil
PUBLISHED: 11-22-2013
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Objective:To explore whether an occupational therapy intervention combined with physiotherapy rehabilitation improved hip fracture patient outcomes regarding emotional distress, fatigue, independence and function.Design:Randomized controlled trial.Setting:Inpatient trauma ward in a rehabilitation and trauma hospital.Participants:One hundred and twenty-two patients admitted into hospital for hip fracture.Intervention:Patients were randomly assigned to a standard care group (SC, n = 61) or a combined treatment group (CT, n = 61). The SC group received conventional hospital care for hip fracture patients and the CT group underwent occupational therapy as well.Main measures:Patients emotional distress (GHQ-28), perceived fatigue (the first item of the BASDAI using a 0-100 visual analogue scale scale), level of independence (Modified Barthel Index) and function (Harris Hip Score) were measured at baseline and one, three and six months after the intervention.Results:Patients in the CT group experienced a considerable decrease of emotional distress at three and six months (p = 0.005 and p < 0.001, respectively). A between-group analysis showed significant differences in emotional distress at one, three and six months (p < 0.001). Although fatigue levels decreased in the SC group, the most significant decline was reported by the CT group at six months (p < 0.001, mean difference = 14 points). Regarding independence level, significant differences were found within groups at each stage, but also between groups at one month in favor of the CT group. Function improved in both groups compared with baseline (p < 0.001), but no significant differences were found in functionality between groups.Conclusion:Although both groups reported significant improvements, patients in the CT group had better scores in emotional distress and dependence throughout follow-up and better scores in all measures at six months.
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Erythema nodosum as azathioprine hypersensitivity reaction in a patient with bullous pemphigoid.
Indian J Dermatol
PUBLISHED: 10-02-2013
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A 65-year-old woman with bullous pemphigoid presented with fever and several red-purple nodular subcutaneous lesions on both lower legs 1 week after starting treatment with azathioprine (AZA). Biopsy of a skin nodule was compatible with erythema nodosum (EN) and hypersensitivity reaction to AZA was suspected. AZA was subsequently discontinued, observing complete remission of fever and EN within 2 weeks. This case highlights the importance of recognizing EN as a possible manifestation of hypersensitivity reaction to AZA.
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The value of stress perfusion cardiovascular magnetic resonance imaging for patients referred from the adult congenital heart disease clinic: 5-year experience at the Toronto General Hospital.
Cardiol Young
PUBLISHED: 09-18-2013
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Background: Vasodilator stress perfusion cardiovascular magnetic resonance imaging is a clinically useful tool for detection of clinically significant myocardial ischaemia in adults. We report our 5-year retrospective experience with perfusion cardiovascular magnetic resonance in a large, quarternary adult congenital heart disease centre. Methods: We reviewed all cases of perfusion cardiovascular magnetic resonance in patients referred from the adult congenital heart disease service. Dipyridamole stress perfusion cardiovascular magnetic resonance was undertaken on commercially available 1.5 and 3 T cardiovascular magnetic resonance scanners. Late gadolinium enhancement imaging was performed 8-10 minutes after completion of the rest perfusion sequence. Navigator whole-heart coronary magnetic resonance angiography was also performed where feasible. Results of stress cardiovascular magnetic resonance were correlated with complementary imaging studies, surgery, and clinical outcomes. Results: Over 5 years, we performed 34 stress perfusion cardiovascular magnetic resonance examinations (11 positive). In all, 84% of patients had further investigations for ischaemia in addition to cardiovascular magnetic resonance. Within a subgroup of 19 patients who had definitive alternative assessment of their coronary arteries, stress perfusion cardiovascular magnetic resonance demonstrated a sensitivity of 82% and specificity of 100%. Of the 34 studies, two were false negatives, in which the aetiology of ischaemia was extrinsic arterial compression rather than intrinsic coronary luminal narrowing. Coronary abnormalities were identified in 71% of cases who had coronary magnetic resonance angiography. Conclusion: Stress perfusion cardiovascular magnetic resonance is a useful and accurate tool for investigation of myocardial ischaemia in an adult congenital heart disease population with suspected non-atherosclerotic coronary abnormalities.
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Health-related Internet use by lupus patients in southern Spain.
Clin. Rheumatol.
PUBLISHED: 09-16-2013
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Internet has become a widely used tool by patients seeking information on different diseases. The information regarding lupus patients Internet use is scarce. This study aims to explore the attitudes and practices of lupus patients in southern Spain, regarding Internet use to find health-related information. A survey was carried out including 150 patients from six Andalusian Hospitals. To search for information, 67.3 % of the patients used Internet. The proportion of female Internet users was higher (69.3 vs 46.2 %), particularly those belonging to a patients association (81.8 vs 32.7 %), and are regular users of Internet (80.2 vs 44.4 %); 37.5 % thought the information found in the Internet was of little use or not useful at all, and 58 % of the respondents stated that the information found caused them concern while for 27 %, it was a relief. Most patients preferred the information given by their physicians (63.6 %); 33.9 % considered that the information from both sources was complementary, and 2.5 % preferred the information obtained from the Internet. A percentage of 85.3 of the patients would like their physicians to provide them with information on high-quality sites regarding their illness. Lupus patients make frequent use of the Internet to look for information on their disease. Considering this, and because better-informed patients follow more precisely the indications given by the physician, medical staff should collaborate in the development of high-quality sites for the patient to have additional sources of information.
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Cardiovascular magnetic resonance imaging predictors of pregnancy outcomes in women with coarctation of the aorta.
Eur Heart J Cardiovasc Imaging
PUBLISHED: 09-12-2013
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The aim of this study was to determine associations between aortic morphometry evaluated by cardiovascular magnetic resonance (CMR) and pregnancy outcomes in women with aortic coarctation (CoA).
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An assessment of the importance of subsurface catch basins for Aedes aegypti adult production during the dry season in a neighborhood of Merida, Mexico.
J. Am. Mosq. Control Assoc.
PUBLISHED: 08-08-2013
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We compared the number of adult Aedes aegypti emerging from subsurface catch basins located in the streets against the number of pupae (as a proxy of adults emerging) from the entire container larval habitats found at residential premises within 1 ha of a neighborhood in the Mexican city of Merida during 8 days in the dry season of 2012. Aedes aegypti adults were collected from 60% of the subsurface catch basins. They produced 12 adults/day/ha (95% confidence interval [CI], 6.4 to 17.9), 5 females (95% CI, 2.1 to 7.7), and 7 males (95% CI, 3.8 to 10.7). In contrast, only 7 containers holding water were identified in 30 premises inspected, 1 bucket was positive for Ae. aegypti larvae, but no pupae-positive containers were found. No other mosquito species were found. This study revealed the importance of this type of nonresidential and subterranean aquatic habitat for Ae. aegypti adult production in this neighborhood of Merida during the dry season.
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Change of residence and functional status within three months and one year following hip fracture surgery.
Disabil Rehabil
PUBLISHED: 08-06-2013
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Abstract Purpose: To study the recovery of patients in terms of 18 activities of daily living and change of residence within the year following a hip fracture. Method: This prospective cohort study was carried out in a trauma service of an acute hospital in southern Spain including 159 patients with a hip fracture, 65 years or older and allowed weight-bearing after surgery. Patients or their relatives were interviewed about their residential status and functional level at pre-fracture, three months and one year after surgery, using the Functional Independence Measure. Results: Losses of function for the main activities affected were, at the first month, third month and one year relative to the pre-fracture status, 50%, 25% and 12%, respectively, for locomotion, 40%, 25% and 20%, respectively, for mobility and 27%, 17% and 15%, respectively, for self care (p?
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MicroRNA signatures in hereditary breast cancer.
Breast Cancer Res. Treat.
PUBLISHED: 07-16-2013
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This study aims to identify signatures of miR associated with hereditary, BRCA1 or BRCA2 mutation positive breast cancer (BC), and non-hereditary BC, either sporadic (SBC) or non-informative (BRCAX). Moreover, we search for signatures associated with tumor stage, immunohistochemistry and tumor molecular profile. Twenty formalin fixed paraffin embedded (FFPE) BCs, BRCA1, BRCA2, BRCAX and SBC, five per group were studied. Affymetrix platform miRNA v.3.0 was used to perform miR expression analysis. ER, PR, HER2 and Ki67 protein expression was analyzed by immunohistochemistry. BRCA1, BRCA2 and RASSF1 methylation analysis, AURKA copy number variations, and BRCA1 and BRCA2 deletions, were studied by MLPA. We validated eight of the miR selected by the arrays in 77 BCs by qRT-PCR. The miR profiles associated with tumor features were studied applying the Sparse Partial Least Squares Discriminant Analysis. MiR discrimination capability to distinguish hereditary and non-hereditary BC was analyzed by the discriminant function. With 15 out of 1,733 hsa-miRs, it was possible to differentiate the four groups. BRCA1, BRCA2 and SBC were associated with clusters of hyper-expressed miRs, and BRCAX with hypo-expressed miRs. Hsa-miR-4417 and hsa-miR-423-3p expressions (included among the eight validated miRs) differentiated 70.1 % of hereditary and non-hereditary BCs. We found miR profiles associated with tumor features like node involvement, histological grade, ER, PR and HER2 expression. Regarding molecular parameters, we only found a weak association of miRs in BC harboring losses in AURKA. We conclude that array miR expression profiles can differentiate the four study groups using FFPE BC. However, miRs expression estimated by qRT-PCR differentiates only hereditary and non-inherited BCs. The miR expression array is a simple and rapid approach that could be useful to facilitate the identification of those SBC carrying genetic or epigenetic changes in BRCA genes responsible of BRCA-like phenotype. These patients could benefit from the treatment with PARP inhibitors.
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Anomalous protein-DNA interactions behind neurological disorders.
Adv Protein Chem Struct Biol
PUBLISHED: 06-25-2013
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Aggregation, nuclear location, and nucleic acid interaction are common features shared by a number of proteins related to neurodegenerative diseases, including Alzheimers disease, Parkinsons disease, amyotrophic lateral sclerosis, transmissible spongiform encephalopathy, Huntingtons disease, spinobulbar muscular atrophy, dentatorubro-pallidoluysian atrophy, and several spinocerebellar ataxias. ?-Amyloid peptides, tau protein, ?-synuclein, superoxide dismutase1, prion protein, huntingtin, atrophin1, androgen receptor, and several ataxins are proteins prone to becoming aggregated, to translocate inside cell nucleus, and to bind DNA. In this chapter, we review those common features suggesting that neurological diseases too may share a transcriptional disorder, making it an important contribution to the origin of the disease.
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Association of cyclists age and sex with risk of involvement in a crash before and after adjustment for cycling exposure.
Accid Anal Prev
PUBLISHED: 06-12-2013
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This study aimed to estimate the association of cyclists age and sex with the risk of being involved in a crash with and without adjustment for their amount of exposure. We used the distribution of the entire population and cyclists (total and non-responsible) involved in road crashes in Spain between 1993 and 2009 held by the Spanish National Institute of Statistics and the Spanish General Traffic Directorate to calculate rates of exposure and involvement in a crash. Males aged 45-49 years were used as the reference category to obtain exposure rate ratios (ERR) and unadjusted crash rate ratios (URR). We then used these values in decomposition analysis to calculate crash rate ratios adjusted for exposure (ARR). The pattern of ARR was substantially different from URR. In both sexes the highest values were observed in the youngest age groups, and the values decreased as age increased except for a slight increase in the oldest age groups. In males, a slight increase in the lowest and highest age categories was observed for crashes resulting in severe injury or death, and a decrease was observed for the youngest cyclists who were wearing a helmet. The large differences between age and sex groups in the risk of involvement in a cycling crash are strongly dependent on differences in their exposure rates. Taking exposure rates into account, cyclists younger than 30 years and older than 65 years of age had the highest risk of being involved in a crash.
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Mediterranean diet, kidney function, and mortality in men with CKD.
Clin J Am Soc Nephrol
PUBLISHED: 06-06-2013
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Adherence to a Mediterranean diet may link to a better preserved kidney function in the community as well as a favorable cardiometabolic profile and reduced mortality risk in individuals with manifest CKD.
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Image noise-based dose adaptation in dynamic volume CT of the heart: dose and image quality optimisation in comparison with BMI-based dose adaptation.
Eur Radiol
PUBLISHED: 06-01-2013
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To compare the image quality and radiation dose using image-noise (IN)-based determination of X-ray tube settings compared with a body mass index (BMI)-based protocol during CT coronary angiography (CTCA).
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Simultaneous phenotypic and genetic characterization of single circulating tumor cells from colon cancer patients.
Histol. Histopathol.
PUBLISHED: 05-08-2013
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Since circulating tumor cells (CTCs) have metastatic potential, their genetic and phenotypic characteristics could provide crucial information to establish the most effective therapy. We assessed the clinical utility of a methodology that allows the simultaneous analysis of CTC phenotype and genotype in colon cancer patients and, in addition, whether this methodology could provide complementary information to that obtained by the primary tumor biopsy. Thirty-three non-metastatic (stages 0-III) colon cancer patients and 9 healthy donor samples were evaluated. All peripheral blood samples (10 ml) were analyzed by cytokeratin immunomagnetic enrichment. Eight samples were analyzed by immunocytochemistry and 25 samples were analyzed by FICTION technique for simultaneous cytokeratin expression and chromosome 17 and ERBB2 gene status. A further study was carried out in one patient who showed CTC heterogeneity in chromosomal abnormalities. We analyzed HER2 protein expression on CTCs and FISH and HER2 protein expression in primary tumor of this patient. Our results show that 9.09% of patients had cytokeratin-positive CTCs (CK+/CTCs in peripheral blood). One of the patients showed heterogeneity in chromosomal 17 abnormalities and two different CK expression patterns on CTCs: one CK+/CTCs and one CK-/CTCs. Furthermore, 63.33% of these CTCs overexpressed HER2 protein while the primary tumor of this patient was diploid and did not express HER2 protein. We describe a methodology that allows the simultaneous genetic and phenotypic analysis of CTCs in colon cancer patients, which may provide essential information to select patients who might benefit from specific therapy.
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Simultaneous liquid-liquid extraction and dispersive solid-phase extraction as a sample preparation method to determine acidic contaminants in river water by gas chromatography/mass spectrometry.
Talanta
PUBLISHED: 04-30-2013
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A sample preparation procedure that combines a liquid-liquid extraction (LLE) with a dispersive solid-phase extraction (DSPE) has been devised to determine residues of four phenoxyacid herbicides, two aminopolycarboxylic acids and five acidic anti-inflammatory drugs in small volumes of river water samples. Two aliquots of acetone (3 and 0.5 mL) were used to extract the analytes from a 10 mL water sample at pH 2 containing 5 mg of octadecylsilane (ODS) sorbent and NaCl at a 5.5 M concentration. Acetone was isolated by the salting-out effect, collected, evaporated and the extract was treated with BF3/methanol to obtain the methyl esters of the analytes and determine them by GC with mass spectrometric detection. Recoveries were comprised between 82% and 114% with relative standard deviations about 5-15% (n=5) within a concentration range about 0.03-44 µg L(-1). The amount of ODS added to sample resulted to be a significant factor in the recovery for most of the analytes as deduced from an experimental design; the sample pH was not a so critical factor. A robustness study of the proposed sample preparation was carried out as defined by Youden and Steiner and an estimation of the uncertainties of the measured concentrations was made following the EURACHEM/CITAC guidelines, too.
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Is heparin reversal required for the safe performance of percutaneous endovascular aortic aneurysm repair?
Ann Vasc Surg
PUBLISHED: 04-09-2013
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Percutaneous endovascular aneurysm repair (PEVAR) can be performed with high technical success rates and low morbidity rates. Several peer-reviewed papers regarding PEVAR have routinely combined heparin reversal with protamine before sheath removal. The risks of protamine reversal are well documented and include cardiovascular collapse and anaphylaxis. The aim of this study is to review outcomes of patients who underwent PEVAR without heparin reversal.
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The proposed changes for DSM-5 for SLD and ADHD: international perspectives--Australia, Germany, Greece, India, Israel, Italy, Spain, Taiwan, United Kingdom, and United States.
J Learn Disabil
PUBLISHED: 04-06-2013
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This article presents an international perspective of the proposed changes to the DSM-5 for learning disabilities (LD) and attention-deficit/hyperactivity disorders (ADHD) across ten countries: Australia, Germany, Greece, India, Israel, Italy, Spain, Taiwan, the United Kingdom, and the United States. We provide perspectives of the present situation for youth with LD and youth with ADHD and describe the legislation, prevalence rates, and educational systems that serve students with disabilities in the respective countries. We also present a discussion of the expected impact of the proposed changes for the diagnosis of LD and ADHD in each country.
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Multimodality imaging for resuscitated sudden cardiac death.
J Cardiovasc Med (Hagerstown)
PUBLISHED: 03-16-2013
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We present a case that elegantly illustrates the utility of two novel noninvasive imaging techniques, computed tomography (CT) coronary angiography and cardiac MRI, in the diagnosis and management of a 27-year-old man with exertion-induced cardiac arrest caused by an anomalous right coronary artery. CT coronary angiography with 3D reformatting delineated the interarterial course of an anomalous right coronary artery compressed between the aorta and pulmonary artery, whereas cardiac MRI showed a small myocardial infarction in the right coronary artery territory not detected on echocardiography. This case highlights the value of novel multimodality imaging techniques in the risk stratification and management of patients with resuscitated cardiac arrest.
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Novel and recurrent BRCA1/BRCA2 mutations in early onset and familial breast and ovarian cancer detected in the Program of Genetic Counseling in Cancer of Valencian Community (eastern Spain). Relationship of family phenotypes with mutation prevalence.
Fam. Cancer
PUBLISHED: 03-13-2013
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During the first 6 years of the Program of Genetic Counselling in Cancer of Valencia (eastern Spain), 310 mutations (155 in BRCA1 and 155 in BRCA2) in 1,763 hereditary breast (BC) and ovarian cancer (OC) families were identified. Of the mutations found 105 were distinct (53 in BRCA1 and 52 in BRCA2), eight new and 37 recurrent. Two of the novel mutations were frame-shift placed in exons 2 and 11 of BRCA1 and the remaining six were placed in BRCA2; four frame-shift (three in exon 11 and one in exon 23), one deletion of the entire exon 19 and one in the intervening sequence of exon 22. The BRCA1 mutations with higher recurrence were c.66_68delAG, c.5123C > A, c.1961delA, c.3770_3771delAG and c.5152+5G > A that covered 45.2% of mutations of this gene. The age of onset of BCs of c.68_69delAG mutation carriers occurs later than for the other recurrent mutations of this gene (45 vs. 37 years; p = 0.008). The BRCA2 mutations with higher recurrence were c.9026_9030delATCAT, c.3264insT and c.8978_8991del14 which represented 43.2% of all mutations in this gene, being the most recurrent mutation by far c.9026_9030delATCAT that represents 21.3% of BRCA2 mutations and 10.6% of all mutations. Probands with family histories of BC and OC, or OC and/or BC in at least two first degree relatives, were the more likely to have BRCA1/BRCA2 mutations (35.2% of the total mutations). And that most BRCA1mutations (73.19% mutations) occurred in probands with early-onset BC or with family history of OC.
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Excess podocyte semaphorin-3A leads to glomerular disease involving plexinA1-nephrin interaction.
Am. J. Pathol.
PUBLISHED: 03-07-2013
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Semaphorin-3A (Sema3a), a guidance protein secreted by podocytes, is essential for normal kidney patterning and glomerular filtration barrier development. Here, we report that podocyte-specific Sema3a gain-of-function in adult mice leads to proteinuric glomerular disease involving the three layers of the glomerular filtration barrier. Reversibility of the glomerular phenotype upon removal of the transgene induction provided proof-of-principle of the cause-and-effect relationship between podocyte Sema3a excess and glomerular disease. Mechanistically, excess Sema3a induces dysregulation of nephrin, matrix metalloproteinase 9, and ?v?3 integrin in vivo. Sema3a cell-autonomously disrupts podocyte shape. We identified a novel direct interaction between the Sema3a signaling receptor plexinA1 and nephrin, linking extracellular Sema3a signals to the slit-diaphragm signaling complex. We conclude that Sema3a functions as an extracellular negative regulator of the structure and function of the glomerular filtration barrier in the adult kidney. Our findings demonstrate a crosstalk between Sema3a and nephrin signaling pathways that is functionally relevant both in vivo and in vitro.
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Coronary calcium scan acquisition before coronary CT angiography: limited benefit or useful addition?
AJR Am J Roentgenol
PUBLISHED: 02-21-2013
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This article reviews the role of coronary calcium quantification in symptomatic patients and the pros and cons of acquiring an unenhanced coronary calcium scan in every patient with suspected coronary artery disease referred for coronary CT angiography.
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Lower than expected cytogenetic and molecular response to imatinib in Mexican patients with chronic myelogenous leukemia.
Hematology
PUBLISHED: 02-20-2013
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Imatinib has been considered as the gold standard for drug therapy of chronic myelogenous leukemia (CML) because it offers higher cytogenetic response and better quality of life than traditional drugs. In this study we applied the standard 400 mg dose of imatinib in 37 CML Ph (+) Mexican patients, monitoring their cytogenetic response using fluorescent in situ hybridization and carrying out molecular analyses using reverse transcription polymerase chain reaction. The study included 19 male and 18 female patients with a median age of 41 years. The median follow-up time from diagnosis was 56 months. Thirty-six patients (97%) achieved complete hematologic response in a median time of 29 days. Complete cytogenetic response and complete molecular remission was observed in only five (13%) and three (8.1%) patients, respectively, less than the expected rate (50-90%) reported in other studies.
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Interruption of transmission of Onchocerca volvulus in the Southern Chiapas Focus, México.
PLoS Negl Trop Dis
PUBLISHED: 02-09-2013
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The Southern Chiapas focus of onchocerciasis in Southern Mexico represents one of the major onchocerciasis foci in Latin America. All 559 endemic communities of this focus have undergone semi-annual mass treatment with ivermectin since 1998. In 50 communities of this focus, ivermectin frequency shifted from twice to four times a year in 2003; an additional 113 communities were added to the quarterly treatment regimen in 2009 to achieve a rapid suppression of transmission.
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The type VI secretion system encoded in Salmonella pathogenicity island 19 is required for Salmonella enterica serotype Gallinarum survival within infected macrophages.
Infect. Immun.
PUBLISHED: 01-28-2013
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Salmonella enterica serotype Gallinarum is the causative agent of fowl typhoid, a disease characterized by high morbidity and mortality that causes major economic losses in poultry production. We have reported that S. Gallinarum harbors a type VI secretion system (T6SS) encoded in Salmonella pathogenicity island 19 (SPI-19) that is required for efficient colonization of chicks. In the present study, we aimed to characterize the SPI-19 T6SS functionality and to investigate the mechanisms behind the phenotypes previously observed in vivo. Expression analyses revealed that SPI-19 T6SS core components are expressed and produced under in vitro bacterial growth conditions. However, secretion of the structural/secreted components Hcp1, Hcp2, and VgrG to the culture medium could not be determined, suggesting that additional signals are required for T6SS-dependent secretion of these proteins. In vitro bacterial competition assays failed to demonstrate a role for SPI-19 T6SS in interbacterial killing. In contrast, cell culture experiments with murine and avian macrophages (RAW264.7 and HD11, respectively) revealed production of a green fluorescent protein-tagged version of VgrG soon after Salmonella uptake. Furthermore, infection of RAW264.7 and HD11 macrophages with deletion mutants of SPI-19 or strains with genes encoding specific T6SS core components (clpV and vgrG) revealed that SPI-19 T6SS contributes to S. Gallinarum survival within macrophages at 20 h postuptake. SPI-19 T6SS function was not linked to Salmonella-induced cytotoxicity or cell death of infected macrophages, as has been described for other T6SS. Our data indicate that SPI-19 T6SS corresponds to a novel tool used by Salmonella to survive within host cells.
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Determination of aminopolycarboxylic acids in river water by solid-phase extraction on activated charcoal cartridges and gas chromatography with mass spectrometric detection. Method performance characteristics and estimation of the uncertainty.
Anal. Chim. Acta
PUBLISHED: 01-27-2013
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A new sample preparation procedure to determine aminopolycarboxylic acids (ethylenediaminetetraacetic acid, EDTA, nitrilotriacetic acid, NTA, diethylenetriaminepentaacetic acid, DTPA, and cyclohexanediaminetetraacetic acid, CDTA) in river water is described. The procedure consists of the solid-phase extraction of the aminopolycaroxyllic acids on activated charcoal cartridges after increasing the ionic strength and acidifying the sample. The extract was eluted with methanol and the analytes were methylated in presence of BF3/methanol to determine them by GC with mass spectrometric detection. Recoveries were higher than 90% with good repeatabilities and inter-day precision for concentrations close to quantification limits (about 10 ?g L(-1)) and higher. It has been verified that the proposed method is robust according to the Youden and Steiner test and free of matrix effects arisen from the presence of organic matter and iron(III) as deduced from statistical tests. A bottom-up approach was followed to estimate the uncertainty of the measured concentration. At concentrations close to 10 ?g L(-1) the most relevant step of the method is the calculus of the interpolated concentration which has a high value of relative standard uncertainty.
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Towards a better use of psychoanalytic concepts: a model illustrated using the concept of enactment.
Int J Psychoanal
PUBLISHED: 01-16-2013
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It is well known that there is a lack of consensus about how to decide between competing and sometimes mutually contradictory theories, and how to integrate divergent concepts and theories. In view of this situation the IPA Project Committee on Conceptual Integration developed a method that allows comparison between different versions of concepts, their underlying theories and basic assumptions. Only when placed in a frame of reference can similarities and differences be seen in a methodically comprehensible and reproducible way. We used "enactment" to study the problems of comparing concepts systematically. Almost all psychoanalytic schools have developed a conceptualization of it. We made a sort of provisional canon of relevant papers we have chosen from the different schools. The five steps of our method for analyzing the concept of enactment will be presented. The first step is the history of the concept; the second the phenomenology; the third a methodological analysis of the construction of the concept. In order to compare different conceptualizations we must know the main dimensions of the meaning space of the concept, this is the fourth step. Finally, in step five we discuss if and to what extent an integration of the different versions of enactment is possible.
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Cloning, production, and functional expression of the bacteriocin sakacin A (SakA) and two SakA-derived chimeras in lactic acid bacteria (LAB) and the yeasts Pichia pastoris and Kluyveromyces lactis.
J. Ind. Microbiol. Biotechnol.
PUBLISHED: 01-09-2013
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Mature sakacin A (SakA, encoded by sapA) and its cognate immunity protein (SakI, encoded by sapiA), and two SakA-derived chimeras mimicking the N-terminal end of mature enterocin P (EntP/SakA) and mature enterocin A (EntA/SakA) together with SakI, were fused to different signal peptides (SP) and cloned into the protein expression vectors pNZ8048 and pMG36c for evaluation of their production and functional expression by different lactic acid bacteria. The amount, antimicrobial activity, and specific antimicrobial activity of SakA and its chimeras produced by Lactococcus lactis subsp. cremoris NZ9000 depended on the SP and the expression vector. Only L. lactis NZ9000 (pNUPS), producing EntP/SakA, showed higher bacteriocin production and antimicrobial activity than the natural SakA-producer Lactobacillus sakei Lb706. The lower antimicrobial activity of the SakA-producer L. lactis NZ9000 (pNUS) and that of the EntA/SakA-producer L. lactis NZ9000 (pNUAS) could be ascribed to secretion of truncated bacteriocins. On the other hand, of the Lb. sakei Lb706 cultures transformed with the pMG36c-derived vectors only Lb. sakei Lb706 (pGUS) overproducing SakA showed a higher antimicrobial activity than Lb. sakei Lb706. Finally, cloning of SakA and EntP/SakA into pPICZ?A and pKLAC2 permitted the production of SakA and EntP/SakA by recombinant Pichia pastoris X-33 and Kluyveromyces lactis GG799 derivatives although their antimicrobial activity was lower than expected from their production.
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Oxidized single-walled carbon nanohorns as sorbent for porous hollow fiber direct immersion solid-phase microextraction for the determination of triazines in waters.
Anal Bioanal Chem
PUBLISHED: 01-09-2013
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This paper evaluates the potential of oxidized single-walled carbon nanohorns (o-SWNHs) immobilized on the pores of a hollow fiber (HF) for the direct immersion solid-phase microextraction of triazines from waters. The fabrication of the device requires the oxidation of the nanoparticles by means of microwave irradiation in order to obtain a homogeneous dispersion in methanol. Then, a porous hollow fiber is immersed in the methanolic dispersion of the o-SWNHs under ultrasound stirring. This procedure permits the immobilization of the o-SWNHs in the pores of the hollow fiber. For the extraction, a stainless steel wire was introduced inside the fiber to allow the vertical immersion of the o-SWNHs-HF in the aqueous standard/water sample. The triazines were preconcentrated on the immobilized o-SWNHs and further eluted using 150 ?L of methanol. The solvent was evaporated and the residue reconstituted in 10 ?L of methanol for sensitivity enhancement. Gas chromatography-mass spectrometry was selected as instrumental technique. The limits of detection were between 0.05 and 0.1 ?g L(-1) with an excellent precision (expressed as relative standard deviation) between runs (below 10.2 %) and between fibers (below 12.8 %). Finally, the method was applied to the determination of the triazines in fortified waters, an average recovery value of 90 % being obtained.
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Specific binding of DNA to aggregated forms of Alzheimers disease amyloid peptides.
Int. J. Biol. Macromol.
PUBLISHED: 01-06-2013
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Anomalous protein aggregation is closely associated to age-related mental illness. Extraneuronal plaques, mainly composed of aggregated amyloid peptides, are considered as hallmarks of Alzheimers disease. According to the amyloid cascade hypothesis, this disease starts as a consequence of an abnormal processing of the amyloid precursor protein resulting in an excess of amyloid peptides. Nuclear localization of amyloid peptide aggregates together with amyloid-DNA interaction, have been repeatedly reported. In this paper we have used surface plasmon resonance and electron microscopy to study the structure and behavior of different peptides and proteins, including ?-lactoglobulin, bovine serum albumin, myoglobin, histone, casein and the amyloid-? peptides related to Alzheimers disease A?25-35 and A?1-40. The main purpose of this study is to investigate whether proneness to DNA interaction is a general property displayed by aggregated forms of proteins, or it is an interaction specifically related to the aggregated forms of those particular proteins and peptides related to neurodegenerative diseases. Our results reveal that those aggregates formed by amyloid peptides show a particular proneness to interact with DNA. They are the only aggregated structures capable of binding DNA, and show more affinity for DNA than for other polyanions like heparin and polyglutamic acid, therefore strengthening the hypothesis that amyloid peptides may, by means of interaction with nuclear DNA, contribute to the onset of Alzheimers disease.
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Functional Characterization of a Novel Frameshift Mutation in the C-terminus of the Nav1.5 Channel Underlying a Brugada Syndrome with Variable Expression in a Spanish Family.
PLoS ONE
PUBLISHED: 01-01-2013
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We functionally analyzed a frameshift mutation in the SCN5A gene encoding cardiac Na(+) channels (Nav1.5) found in a proband with repeated episodes of ventricular fibrillation who presented bradycardia and paroxysmal atrial fibrillation. Seven relatives also carry the mutation and showed a Brugada syndrome with an incomplete and variable expression. The mutation (p.D1816VfsX7) resulted in a severe truncation (201 residues) of the Nav1.5 C-terminus.
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Usefulness of bladder-prostate ultrasound in the diagnosis of obstruction/hyperactivity in males with BPH.
Arch. Esp. Urol.
PUBLISHED: 12-14-2011
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To determine the utility of prostate ultrasound in the diagnosis of infravesical obstruction (IVO) and detrusor hyperactivity(DH).
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Isolation of UmRrm75, a gene involved in dimorphism and virulence of Ustilago maydis.
Microbiol. Res.
PUBLISHED: 10-21-2011
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Ustilago maydis displays dimorphic growth, alternating between a saprophytic haploid yeast form and a filamentous dikaryon, generated by mating of haploid cells and which is an obligate parasite. Induction of the dimorphic transition of haploid strains in vitro by change in ambient pH has been used to understand the mechanisms governing this differentiation process. In this study we used suppression subtractive hybridization to generate a cDNA library of U. maydis genes up-regulated in the filamentous form induced in vitro at acid pH. Expression analysis using quantitative RT-PCR showed that the induction of two unigenes identified in this library coincided with the establishment of filamentous growth in the acid pH medium. This expression pattern suggested that they were specifically associated to hyphal development rather than merely acid pH-induced genes. One of these genes, UmRrm75, encodes a protein containing three RNA recognition motifs and glycine-rich repeats and was selected for further study. The UmRrm75 gene contains 4 introns, and produces a splicing variant by a 3-alternative splicing site within the third exon. Mutants deleted for UmRrm75 showed a slower growth rate than wild type strains in liquid and solid media, and their colonies showed a donut-like morphology on solid medium. Interestingly, although ?UmRrm75 strains were not affected in filamentous growth induced by acid pH and oleic acid, they exhibited reduced mating, post-mating filamentous growth and virulence. Our data suggest that UmRrm75 is probably involved in cell growth, morphogenesis, and pathogenicity in U. maydis.
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Species diversity, community dynamics, and metabolite kinetics of the microbiota associated with traditional ecuadorian spontaneous cocoa bean fermentations.
Appl. Environ. Microbiol.
PUBLISHED: 09-16-2011
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Traditional fermentations of the local Ecuadorian cocoa type Nacional, with its fine flavor, are carried out in boxes and on platforms for a short time. A multiphasic approach, encompassing culture-dependent and -independent microbiological analyses of fermenting cocoa pulp-bean samples, metabolite target analyses of both cocoa pulp and beans, and sensory analysis of chocolates produced from the respective fermented dry beans, was applied for the investigation of the influence of these fermentation practices on the yeast and bacterial species diversity and community dynamics during cocoa bean fermentation. A wide microbial species diversity was found during the first 3 days of all fermentations carried out. The prevailing ethanol-producing yeast species were Pichia kudriavzevii and Pichia manshurica, followed by Saccharomyces cerevisiae. Leuconostoc pseudomesenteroides (glucose and fructose fermenting), Fructobacillus tropaeoli-like (fructose fermenting), and Lactobacillus fermentum (citrate converting, mannitol producing) represented the main lactic acid bacterial species in the fermentations studied, resulting in intensive heterolactate metabolism of the pulp substrates. Tatumella saanichensis and Tatumella punctata were among the members of the family Enterobacteriaceae present during the initial phase of the cocoa bean fermentations and could be responsible for the production of gluconic acid in some cases. Also, a potential new yeast species was isolated, namely, Candida sorbosivorans-like. Acetic acid bacteria, whose main representative was Acetobacter pasteurianus, generally appeared later during fermentation and oxidized ethanol to acetic acid. However, acetic acid bacteria were not always present during the main course of the platform fermentations. All of the data taken together indicated that short box and platform fermentation methods caused incomplete fermentation, which had a serious impact on the quality of the fermented dry cocoa beans.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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