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Articles by Gary C.T. Chan in JoVE

 JoVE Immunology and Infection

A Quantitative Evaluation of Cell Migration by the Phagokinetic Track Motility Assay

1Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, 2Center for Molecular and Tumor Virology, Louisiana State University Health Sciences Center, 3Department of Microbiology and Immunology, SUNY Upstate Medical University, 4Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center


JoVE 4165

The phagokinetic motility track assay is a method used to assess the movement of cells. Specifically, the assay measures chemokinesis (random cell motility) over time in a quantitative manner. The assay takes advantage of the ability of cells to create a measurable track of their movement on colloidal gold-coated coverslips.

Other articles by Gary C.T. Chan on PubMed

Nasopharyngeal Cancer: EHNS-ESMO-ESTRO Clinical Practice Guidelines for Diagnosis, Treatment and Follow-up

Methods for High-throughput MethylCap-Seq Data Analysis

Advances in whole genome profiling have revolutionized the cancer research field, but at the same time have raised new bioinformatics challenges. For next generation sequencing (NGS), these include data storage, computational costs, sequence processing and alignment, delineating appropriate statistical measures, and data visualization. Currently there is a lack of workflows for efficient analysis of large, MethylCap-seq datasets containing multiple sample groups.

Dual-action Inhibitors of HIF Prolyl Hydroxylases That Induce Binding of a Second Iron Ion

Inhibition of the hypoxia-inducible factor (HIF) prolyl hydroxylases (PHD or EGLN enzymes) is of interest for the treatment of anemia and ischemia-related diseases. Most PHD inhibitors work by binding to the single ferrous ion and competing with 2-oxoglutarate (2OG) co-substrate for binding at the PHD active site. Non-specific iron chelators also inhibit the PHDs, both in vitro and in cells. We report the identification of dual action PHD inhibitors, which bind to the active site iron and also induce the binding of a second iron ion at the active site. Following analysis of small-molecule iron complexes and application of non-denaturing protein mass spectrometry to assess PHD2·iron·inhibitor stoichiometry, selected diacylhydrazines were identified as PHD2 inhibitors that induce the binding of a second iron ion. Some compounds were shown to inhibit the HIF hydroxylases in human hepatoma and renal carcinoma cell lines.

Of Veins, Valves, and Vascular Access!

Is There a Role for Empiric Gastroduodenal Artery Embolization in the Management of Patients with Active Upper GI Hemorrhage?

PURPOSE: To assess the relative efficacy of empiric gastroduodenal artery (GDA) embolization in reducing recurrent hemorrhage compared to image-guided targeted embolization. METHODS: Data were retrospectively collected for consecutive patients who had catheter angiography for major upper gastrointestinal hemorrhage from May 2008 to November 2010 (n = 40). The total number of cases were divided into two main groups according to angiographic findings: those that demonstrated a site of hemorrhage on catheter angiography (group 1, n = 13), and those where the site of hemorrhage was not identified on catheter angiography (group 2, n = 27). Group 2 was then further divided into patients who received empiric embolization (group 2a, n = 20) and those who had no embolization performed after angiography (group 2b, n = 7). RESULTS: The technical and clinical success rates for embolization in groups 1 and 2a were, respectively, 100 vs. 95 %, and 85 vs. 80 %. There was no statistical significance in the recurrent hemorrhage rate, reintervention rate, or 30 day mortality between targeted and empiric embolization groups. There were no complications attributed to embolization within this study cohort. CONCLUSION: Cases of duodenal-related major upper gastrointestinal hemorrhage where no embolization is performed have poor outcome. Empiric embolization of the GDA in patients with major upper gastrointestinal hemorrhage refractory to endoscopic treatment appears to be a safe and effective treatment, with low reintervention rates and good clinical outcome comparable to patients where the site of hemorrhage is localized and embolized with computed tomographic angiography or catheter angiography and embolized.

Novel Mutations in GB and GH Circumvent the Requirement for Known GD Receptors in HSV-1 Entry and Cell-to-cell Spread

Both HSV entry and cell-to-cell spread involve a cascade of cooperative interactions among the essential glycoproteins gD, gB and gH/gL initiated by the binding of gD to a cognate HSV entry receptor. Previously we reported that a variant of glycoprotein B (gB:NT) enabled primary virus entry into cells that are devoid of typical HSV entry receptors. Here, we compare the activities of the gB:NT variant with those of a newly selected variant of glycoprotein H (gH:KV) and a frequently co-selected gB variant, gB:S668N. In combination, gH:KV and gB:S668N enabled primary virus entry as efficiently as gB:NT into cells that lack established HSV entry receptors, but separately each variant effected only limited entry. Remarkably, gH:KV uniquely facilitated secondary virus spread between cells that lack canonical entry receptors. Transient expression of the four essential entry glycoproteins revealed that gH:KV, but not gB:NT, induced fusion between cells lacking the standard receptors. Because gD's involvement remained essential for virus spread and cell fusion, we propose that gH:KV mimics a transition state of gH that responds efficiently to weak signals from gD to reach the active state. Computational modeling of the structures of wild-type gH and gH:KV revealed relatively subtle differences that may account for our experimental findings. Our study shows that (i) the dependence of HSV-1 entry and spread on specific gD receptors can be reduced by sequence changes in the downstream effectors gB and gH, and (ii) the relative roles of gB and gH are different in entry and spread.

SOX4 Enables Oncogenic Survival Signals in Acute Lymphoblastic Leukemia

The SOX4 transcription factor mediates early B cell differentiation. Compared to normal pre-B cells, SOX4 promoter regions in Ph(+) ALL cells are significantly hypo-methylated. Loss- and gain-of-function experiments identified SOX4 as a critical activator of PI3K/AKT and MAPK signaling in ALL cells. ChIP experiments confirmed that SOX4 binds to and transcriptionally activates promoters of multiple components within the PI3K-AKT and MAPK signaling pathways. Cre-mediated deletion of SOX4 had little effect on normal pre-B cells but compromised proliferation and viability of leukemia cells, which was rescued by BCL2L1 and constitutively active AKT and p110 PI3K. Consistent with these findings, high levels of SOX4 expression in ALL cells at the time of diagnosis predicted poor outcome in a pediatric clinical trial (COG P9906). Collectively, these studies identify SOX4 as a central mediator of oncogenic AKT/PI3K and MAPK signaling in ALL.

Clinical Spectrum of Exophiala Infections and a Novel Exophiala Species, Exophiala Hongkongensis

We characterized 12 Exophiala strains isolated over a 15-year period to the species level using phenotypic tests and ITS and Rpb1 sequencing and described the clinical spectrum of the 12 patients. Eight patients had nails or skin infections, two had invasive infections and two had colonization of the gastrointestinal tract. ITS and Rpb1 sequencing showed that 11 of the 12 strains were known Exophiala species [E. oligosperma (n=3), E. jeanselmei (n=2), E. lecanii-corni (n=2), E. bergeri (n=1), E. cancerae (n=1), E. dermatitidis (n=1), and E. xenobiotica (n=1)], which included the first reported cases of onychomycosis caused by E. bergeri and E. oligosperma. The twelfth one (HKU32(T)), isolated from the nail clipping of the right big toe from a 68-year old female patient with onychomycosis, possessed unique morphological characteristics distinct from other Exophiala species. It grew very slowly and had velvety colony texture after 28 days, short conidiophores of the same olivaceous color as supporting hyphae, numerous spores and no chlamydospore-like cells. ITS, Rpb1, β-tubulin and β-actin genes sequencing unambiguously showed that HKU32(T) was clustered with, but formed distinct branches from, other Exophiala species. We propose the new species Exophiala hongkongensis to describe this novel fungus.

A Randomised Comparison of InnoScope and Macintosh Laryngoscope in Simulated Difficult Tracheal Intubation in Manikins

We conducted a crossover randomised study to evaluate the performance of a novel optical stylet, the InnoScope, for tracheal intubation in simulated normal and difficult airways. Twenty-five anaesthetists attempted tracheal intubation on a SimMan 3G simulator using the InnoScope first followed by the Macintosh laryngoscope or vice versa. Three airway scenarios were tested: (1) normal airway; (2) difficult airway with swollen pharynx; and (3) limited neck movement. In each scenario, the laryngeal view, duration of and success rate for tracheal intubation were recorded. Compared with the Macintosh laryngoscope, the use of InnoScope increased the percentage of glottic opening seen by 17% in normal airway, 23% in the difficult airway and 32% with limited neck movement, p < 0.01. Despite this better laryngeal view, successful tracheal intubation achieved with the InnoScope (88.0%) was lower than that for the Macintosh laryngoscope (98.7%), p = 0.008. Using the InnoScope, tracheal intubation during the first attempt was only successful in 48% of cases with difficult airway. In this scenario, the median (interquartile range [range]) duration of tracheal intubation was significantly longer with the InnoScope (53 (20-100 [15-120]) s) compared with the Macintosh laryngoscope (27 (20-62 [15-120]) s), p = 0.01. We conclude that an improved laryngeal view with the use of the InnoScope did not translate into better conditions for tracheal intubation.

Evolution and Phylogeny of the Mud Shrimps (Crustacea: Decapoda) Revealed from Complete Mitochondrial Genomes

ABSTRACT: BACKGROUND: The evolutionary history and relationships of the mud shrimps (Crustacea: Decapoda: Gebiidea and Axiidea) are contentious, with previous attempts revealing mixed results. The mud shrimps were once classified in the infraorder Thalassinidea. Recent molecular phylogenetic analyses, however, suggest separation of the group into two individual infraorders, Gebiidea and Axiidea. Mitochondrial (mt) genome sequence and structure can be especially powerful in resolving higher systematic relationships that may offer new insights into the phylogeny of the mud shrimps and the other decapod infraorders, and test the hypothesis of dividing the mud shrimps into two infraorders. RESULTS: We present the complete mitochondrial genome sequences of five mud shrimps, Austinogebia edulis, Upogebia major, Thalassina kelanang (Gebiidea), Nihonotrypaea thermophilus and Neaxius glyptocercus (Axiidea). All five genomes encode a standard set of 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes and a putative control region. Except for T. kelanang, mud shrimp mitochondrial genomes exhibited rearrangements and novel patterns compared to the pancrustacean ground pattern. Each of the two Gebiidea species (A. edulis and U. major) and two Axiidea species (N. glyptocercus and N. thermophiles) share unique gene order specific to their infraorders and analyses further suggest these two derived gene orders have evolved independently. Phylogenetic analyses based on the concatenated nucleotide and amino acid sequences of 13 protein-coding genes indicate the possible polyphyly of mud shrimps, supporting the division of the group into two infraorders. However, the infraordinal relationships among the Gebiidea and Axiidea, and other reptants are poorly resolved. The inclusion of mt genome from more taxa, in particular the reptant infraorders Polychelida and Glypheidea is required in further analysis. CONCLUSIONS: Phylogenetic analyses on the mt genome sequences and the distinct gene orders provide further evidences for the divergence between the two mud shrimp infraorders, Gebiidea and Axiidea, corroborating previous molecular phylogeny and justifying their infraordinal status. Mitochondrial genome sequences appear to be promising markers for resolving phylogenetic issues concerning decapod crustaceans that warrant further investigations and our present study has also provided further information concerning the mt genome evolution of the Decapoda.

Spatial Morphological and Molecular Differences Within Solid Tumors May Contribute to the Failure of Vascular Disruptive Agent Treatments

ABSTRACT: BACKGROUND: Treatment of solid tumors with vascular disrupting agent OXi4503 results in over 90% tumor destruction. However, a thin rim of viable cells persists in the tumor periphery following treatment, contributing to subsequent recurrence. This study investigates inherent differences in the microenvironment of the tumor periphery that contribute to treatment resistance. METHODS: Using a murine colorectal liver metastases model, spatial morphological and molecular differences within the periphery and the center of the tumor that may account for differences in resistance to OXi4503 treatment were investigated. H&E staining and immunostaining were used to examine vessel maturity and stability, hypoxia and HIF1alpha levels, accumulation of immune cells, expression of proangiogenic factors/receptors (VEGF, TGF-beta, b-FGF, and AT1R) and expression of EMT markers (ZEB1, vimentin, E-cadherin and beta-catenin) in the periphery and center of established tumors. The effects of OXi4503 on tumor vessels and cell kinetics were also investigated. RESULTS: Significant differences were found between tumor periphery and central regions, including association of the periphery with mature vessels, higher accumulation of immune cells, increased growth factor expression, minimal levels of hypoxia and increased evidence of EMT. OXi4503 treatment resulted in collapse of vessels in the tumor center; however vasculature in the periphery remained patent. Similarly, tumor apoptosis and proliferation were differentially modulated between centre and periphery after treatment. CONCLUSIONS: The molecular and morphological differences between tumor periphery and center may account for the observed differential resistance to OXi4503 treatment and could provide targets for drug development to totally eliminate metastases.

Knowledge and Practice Among Hong Kong Oncology Nurses in the Management of Chemotherapy-induced Nausea and Vomiting

PURPOSE: To examine nurses' roles in the prevention and management of chemotherapy-induced nausea and vomiting (CINV), and to identify their related educational needs. METHODS: This was a descriptive cross-sectional study with a self-reported survey completed by 103 oncology nurses caring for and administering chemotherapy to cancer patients in the department of oncology in three Hong Kong public hospitals. The survey was developed to identify key issues pertinent to the role of nurses in managing CINV. Data were collected from the following areas (a) demographics, (b) assessment of CINV, (c) CINV management and (d) barriers and facilitators to good CINV practice. RESULTS: Only a third of respondents performed a CINV assessment before starting chemotherapy, and more than 40% reported that the use of a standardised assessment tool was uncommon. Nearly half recognised that they had inadequate knowledge of different aspects of CINV, but the majority could clearly state the most common pharmacological agents used to treat chemotherapy-induced nausea (88.3%) and vomiting (87.4%). The barriers respondents most frequently encountered in CINV prevention and management were lack of time and a heavy workload. Adopting a standardised CINV assessment tool and management protocol together with further professional training were identified as the major facilitators in improving CINV prevention and management. CONCLUSIONS: Respondents perceived their knowledge of CINV prevention and management as inadequate. There is a need to adopt a standardised assessment tool, to develop a management protocol and to introduce further professional training to meet the expanding needs of both patients and nurses.

Hospitalisation Associated with the 2009 H1N1 Pandemic and Seasonal Influenza in Hong Kong, 2005 to 2010

Aberrant Overexpression of IL-15 Initiates Large Granular Lymphocyte Leukemia Through Chromosomal Instability and DNA Hypermethylation

How inflammation causes cancer is unclear. Interleukin-15 (IL-15) is a pro-inflammatory cytokine elevated in human large granular lymphocyte (LGL) leukemia. Mice overexpressing IL-15 develop LGL leukemia. Here, we show that prolonged in vitro exposure of wild-type (WT) LGL to IL-15 results in Myc-mediated upregulation of aurora kinases, centrosome aberrancies, and aneuploidy. Simultaneously, IL-15 represses miR-29b via induction of Myc/NF-κBp65/Hdac-1, resulting in Dnmt3b overexpression and DNA hypermethylation. All this is validated in human LGL leukemia. Adoptive transfer of WT LGL cultured with IL-15 led to malignant transformation in vivo. Drug targeting that reverses miR-29b repression cures otherwise fatal LGL leukemia. We show how excessive IL-15 initiates cancer and demonstrate effective drug targeting for potential therapy of human LGL leukemia.

Prevalence of Papillary Changes and Folliculosis of the Palpebral Conjunctiva in Asymptomatic Chinese Children

PURPOSE: To determine the prevalence of papillary changes of the upper palpebral conjunctiva and folliculosis of the lower palpebral conjunctiva in Chinese children with no history of contact lens wear. METHOD: Ninety-nine subjects (aged 6-15 years old) who were interested in a myopia control study were screened for papillary changes and folliculosis of the palpebral conjunctiva. Photodocumentation was performed under white and blue light (after the application of fluorescein) with a yellow filter and the photographs were graded by a group of practitioners according to a pre-set grading scale. Analysis was performed with the subjects divided into groups according to gender and age. RESULTS: More than 48% of the subjects had clinically significant (≥Grade 3) papillary changes in the upper palpebral conjunctiva. The prevalence of significant folliculosis in the lower lid was about 33%. The prevalence of significant papillary changes and folliculosis were similar between genders. No differences were observed between younger (age≤10 years old) and older (age>10 years old) in papillary changes but younger subjects showed a higher prevalence of folliculosis. CONCLUSIONS: The prevalences of clinically significant papillary changes and folliculosis of unknown aetiology are high in Chinese children.

The Use of High-resolution Melting Analysis for Rapid Spa Typing on Methicillin-resistant Staphylococcus Aureus Clinical Isolates

Methicillin-resistant Staphylococcus aureus (MRSA) has been endemic in Hong Kong for three decades. This study evaluated the practical use of high-resolution melting (HRM) real-time PCR analysis on MRSA staphylococcal Protein A (spa) typing on local MRSA isolates. Among 55 clinical MRSA isolates collected in 2011, 12 different spa types were observed by the conventional PCR-sequencing method including the locally predominant spa type t1081 and two locally predominant community acquired MRSA spa types t019 and t437. By using the HRM method, it could differentiate all 12 spa genotypes by distinct melting curves and HRM difference plot analysis. These two methods demonstrated 100% concordance whereas the HRM method required only 3h of turnaround time and one-fifth of reagent cost compared to the conventional method. Our study confirmed that the cost effective and rapid HRM typing approach is practically useful for MRSA community transmission monitoring and nosocomial outbreak control in Hong Kong.

Standardized Outpatient Management of Klebsiella Pneumoniae Liver Abscesses

OBJECTIVES: Community-acquired Klebsiella pneumoniae has emerged as a major cause of liver abscess in Asia. Using a standardized protocol, we conducted a prospective cohort study of all cases of K. pneumoniae liver abscess treated from 2005 to 2011 at two outpatient parenteral antimicrobial therapy (OPAT) centers in Singapore, to assess the safety and efficacy of treatment. METHODS: We included all OPAT eligible patients with radiologically confirmed (computed tomography or ultrasound) liver abscesses and K. pneumoniae-positive microbiological cultures obtained from abscess fluid and/or blood at two university teaching hospitals. The endpoints investigated were cure, clinical response, readmission, and mortality. RESULTS: All 109 patients enrolled in the study successfully completed treatment in OPAT. Nine patients required a short-term readmission due to clinical deterioration. There were no deaths or relapses at 30 days post cessation of antibiotics. Abscess size greater than 5cm was associated with a delayed clinical response (odds ratio 5.34, 95% confidence interval 1.25-22.91, p=0.02). CONCLUSION: The management of K. pneumoniae liver abscesses via OPAT using a standardized protocol is a safe and effective alternative to inpatient intravenous antibiotics.

The Effect of Hyperbaric Oxygen and Air on Cartilage Tissue Engineering

ABSTRACT: There is an urgent need to develop tissue-engineered cartilage for patients experiencing joint malfunction due to insufficient self-repairing capacity of articular cartilage. The aim of this research was to explore the effect of hyperbaric oxygen and air on tissue-engineered cartilage formation from human adipose-derived stem cells seeding on the gelatin/polycaprolactone biocomposites. The results of histological analyses indicate that under hyperbaric oxygen and air stimulation, the cell number of chondrocytes in cartilage matrix was not significantly increased, but the 1,9-dimethylmethylene blue assay showed that the glycosaminoglycans syntheses markedly increased compared to the control group. In quantification real-time polymerase chain reaction results, the chondrogenic-specific gene expression of SOX9, aggrecan, and COL2A1 were compared respectively. Within the limitation of this study, it was concluded that 2.5 atmosphere absolute oxygen and air may provide a stress environment to help cartilage tissue engineering development.

Adipose-derived Stem Cells Seeded on Acellular Dermal Matrix Grafts Enhance Wound Healing in a Murine Model of a Full-thickness Defect

The promotion of wound healing using dermal substitutes has become increasingly widespread, but the outcomes of substitute-assisted healing remain functionally deficient. Adipose-derived stem cells (ASCs) have been investigated widely in regenerative medicine and tissue engineering, and they have the potential to enhance wound healing. In this study, we focused on investigating the effects and mechanism of ASCs combined with an acellular dermal matrix (ADM) to treat full-thickness cutaneous wounds in a murine model.

Association and Prognostic Value of Serum Inflammation Markers in Patients with Leukoplakia and Oral Cavity Cancer

Abstract Background: Oral cavity cancer ranks as the fourth leading cancer in men in Taiwan. The development of a serum biomarker panel for early detection and disease monitoring is, therefore, warranted. Methods: Nine inflammation-associated markers were investigated in 46 patients with leukoplakia, 151 patients with untreated oral cavity squamous cell carcinoma (OSCC), and 111 age- and gender-matched healthy controls using enzyme-linked immunosorbent assay. During a subsequent 28-month surveillance of OSCC patients, serum samples were prospectively collected at predetermined intervals following the completion of therapy. Results: Logistic regression analysis showed matrix metalloproteases (MMP)-2, MMP-9, C-reactive protein (CRP), transforming growth factor-β1 (TGF-β1), and E-selectin having the best discrimination power between groups and significant elevation trends of those five markers were noted from control to OSCC. By combining those five markers, a 0.888 and 0.938 area under curve by ROC curve analysis with 67.4% and 80% overall sensitivity and fixed 90% specificity for leukoplakia and OSCC groups were demonstrated. In the follow-up period, 25 OSCC patients developed recurring or secondary tumors. All examined markers had decreased in relapse-free patients following treatment. However, in patients with relapse, interleukin-6, CRP, and serum amyloid A remained at elevated levels. Statistical analysis showed that patients with CRP ≧2 mg/L and E-selectin ≧85 ng/mL at baseline had highest probability of relapse (odds ratio=3.029, p<0.05). Conclusions: The results indicate that inflammation plays a crucial role in the pathogenesis process of OSCC. By examining the inflammation markers, physicians could potentially identify patients at risk of cancer transformation or relapse.

Wnt Signaling Promotes Muller Cell Proliferation and Survival After Injury

PURPOSE: Muller glia respond to retinal injury by a reactive gliosis but only rarely do mammalian glial cells re-enter the cell cycle and generate new neurons. In the non-mammalian retina, however, Muller glia act as stem/progenitor cells. Here, we test the function of Wnt signaling in the post-injury retina, focusing on its ability to influence mammalian Muller cell de-differentiation, proliferation and neurogenesis. METHODS: A Nd:YAG laser was used to create light burns on the retina of Axin2(LacZ/+) Wnt reporter mice. At various timepoints after injury, retinas were analyzed for evidence of Wnt signaling as well as glial cell response, proliferation, and apoptosis. Laser injuries were also created in Axin2(LacZ/LacZ) mice, and the effect of potentiated Wnt signaling on retinal repair was assessed. RESULTS: A subpopulation of mammalian Muller cells are Wnt responsive and when Wnt signaling is increased these cells showed enchanced proliferation in response to injury. In an environment of heightened Wnt signaling, caused by the loss of Wnt negative regulator Axin2, Muller cells proliferate after injury and adopted the expression patterns of retinal progenitor cells (RPCs). The Wnt-responsive Muller cells also exhibited long-term survival and in some cases, expressed the rod photoreceptor marker, Rhodopsin. CONCLUSIONS: The Wnt pathway is activated by retinal injury, and prolonging the endogenous Wnt signal causes a subset of Muller cells to proliferate and de-differentiate into RPCs. These data raise the possibility that transient amplification of Wnt signaling after retinal damage may unlock the latent regenerative capacity long speculated to reside in mammalian neural tissues.

Association Between Smoking and Size of Anal Warts in HIV-infected Women

While the association between smoking and human papillomavirus infection, cervical cancer, and anal cancer has been well studied, evidence on the association between cigarette smoking and anal warts is limited. The purpose of this study was to investigate if cigarette smoking status influences the size of anal warts over time in HIV-infected women in a sample of 976 HIV-infected women from the Women's Interagency HIV Study (WIHS). A linear mixed model was used to determine the effect of smoking on anal wart size. Even though women who were currently smokers had larger anal warts at baseline and slower growth rate of anal wart size after each visit than women who were not current smokers, there was no association between size of anal wart and current smoking status over time. Further studies on the role of smoking and interaction between smoking and other risk factors, however, should be explored.

Studies on Combination of Platinum Drugs Cisplatin and Oxaliplatin with Phytochemicals Anethole and Curcumin in Ovarian Tumour Models

Chemopreventative phytochemicals having antitumour and antioxidant properties can overcome problems of chemoresistance and nonspecific toxicity towards normal cells that are associated with platinum-based chemotherapy against cancer. These agents exert their effects by bringing into play numerous cellular proteins that in turn affect multiple steps in pathways leading to tumourigenesis. In this study, combinations of two cytotoxic phytochemicals anethole and curcumin were applied in binary combination with platinum drugs cisplatin and oxaliplatin to three epithelial ovarian cancer cell lines: A2780 (parent), A2780(cisR) (cisplatin-resistant) and A2780(ZD0473R) (ZD0473-resistant). Cell viability was quantified using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) reduction assay and the combined drug action was analyzed based on the equations derived by Chou and Talalay (1984). Greatest synergism was observed when the phytochemical was added first followed by the platinum drug 2 h later and additiveness to antagonism in combined drug action was observed when the two compounds were administered as a bolus. If confirmed in vivo, the appropriate sequenced combinations of platinum with the phytochemicals may provide a means of overcoming drug resistance.

Epigallocatechin Gallate Acts Synergistically in Combination with Cisplatin and Designed Trans-palladiums in Ovarian Cancer Cells

In this study, synergism in activity from the sequenced combinations of three trans-palladiums (denoted as TH5, TH6 and TH7) with green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG), as well as that with cisplatin, was investigated in a number of human ovarian tumour models as a function of sequence of administration. Cellular accumulation of platinum and palladium, and the levels of platinum-DNA and palladium-DNA binding were also determined for the 0/4 h and 0/0 h sequences of administration. The results of the study show that co-administration of cisplatin with EGCG (0/0 h) produces weak synergism in both cisplatin-sensitive (A2780) and cisplatin-resistant (A2780(cisR)) cell lines whereas (0/4 h) administration produces pronounced synergism in both. In contrast, bolus administration of EGCG with TH5, TH6 and TH7 produces marked antagonism except that with TH5, in the A2780(cisR) cell line, where a mild synergism is observed. In the case of TH5, TH6 and TH7, administration of drugs with a time gap (0/4 h or 4/0 h combinations) produces sequence-dependent synergism in both A2780 and A2780(cisR) cell lines, whereas in the case of cisplatin, marked antagonism is observed with the 4/0 h sequence of administration in the A2780 cell line. Whereas the highly synergistic 0/4 h sequence of combination of cisplatin with EGCG is found to be associated with pronounced cellular accumulation of platinum and a high level of platinum-DNA binding, no such clear trend can be seen for any of the combinations of TH5, TH6 and TH7 with EGCG. The results of the present study provide support to the idea that sequenced combinations of platinum drugs and tumour-active palladium compounds with selected phytochemicals such as EGCG may provide a means of overcoming drug resistance.

Adiponectin Mediated MHC Class II Mismatched Cardiac Graft Rejection in Mice Is IL-4 Dependent

Adiponectin regulates glucose and fatty-acid metabolism but its role in chronic graft rejection mediated by Th2 cytokines remains ill-defined.

Development of Miniaturized Walking Biological Machines

The quest to 'forward-engineer' and fabricate biological machines remains a grand challenge. Towards this end, we have fabricated locomotive "bio-bots" from hydrogels and cardiomyocytes using a 3D printer. The multi-material bio-bot consisted of a 'biological bimorph' cantilever structure as the actuator to power the bio-bot, and a base structure to define the asymmetric shape for locomotion. The cantilever structure was seeded with a sheet of contractile cardiomyocytes. We evaluated the locomotive mechanisms of several designs of bio-bots by changing the cantilever thickness. The bio-bot that demonstrated the most efficient mechanism of locomotion maximized the use of contractile forces for overcoming friction of the supporting leg, while preventing backward movement of the actuating leg upon relaxation. The maximum recorded velocity of the bio-bot was ~236 µm s(-1), with an average displacement per power stroke of ~354 µm and average beating frequency of ~1.5 Hz.

Evidence for Crossmodal Interactions Across Depth on Target Localisation Performance in a Spatial Array

Auditory stimuli are known to improve visual target recognition and detection when both are presented in the same spatial location. However, most studies have focused on crossmodal spatial congruency along the horizontal plane and the effects of audio-visual spatial congruency in depth (i.e., along the depth axis) are relatively less well understood. In the following experiments we presented a visual (face) or auditory (voice) target stimulus in a location on a spatial array which was either spatially congruent or incongruent in depth (i.e., positioned directly in front or behind) with a crossmodal stimulus. The participant's task was to determine whether a visual (experiments 1 and 3) or auditory (experiment 2) target was located in the foreground or background of this array. We found that both visual and auditory targets were less accurately located when crossmodal stimuli were presented from different, compared to congruent, locations in depth. Moreover, this effect was particularly found for visual targets located in the periphery, although spatial incongruency affected the location of auditory targets across both locations. The relative distance of the array to the observer did not seem to modulate this congruency effect (experiment 3). Our results add to the growing evidence for multisensory influences on search performance and extend these findings to the localisation of targets in the depth plane.

The Dollars and Sense of Restraints and Seclusion

Restraints and seclusion are a common practice in many human service settings despite the increasing evidence questioning their efficacy and appropriateness. There are many detrimental effects on people subject to these practices, such as falls, injury, psychological trauma and even death. In addition to the impact on people being served, there is also a range of negative effects on organisations and the workforce. This article outlines and discusses the costs to organisations in implementing restraints and seclusion, and the economic cost-benefits to be gained in working towards the safe elimination of restraints and seclusion. A brief outline of ethical alternatives to restraints and seclusion is explored. The emerging research evidence suggests that it is possible to achieve the safe elimination of restraints and seclusion in a human service organisation.

Ultralow-Threshold Two-Photon Pumped Amplified Spontaneous Emission and Lasing from Seeded CdSe/CdS Nanorod Heterostructures

Ultralow-threshold two-photon pumped amplified spontaneous emission (2ASE) and lasing in seeded CdSe/CdS nanodot/nanorod heterostructures is demonstrated for the first time. Such heterostructures allow the independent tunability of the two-photon absorption (2PA) cross-section (σ(2)) through varying the CdS rod size, and that of the emission wavelength through varying the CdSe dot size. With an enhanced σ(2), 2ASE in these heterostructures is achieved with an ultralow threshold fluence of ∼1.5 mJ/cm(2), which is as much as one order less than that required for spherical semiconductor NCs. Importantly, by exploiting this unique property of the seeded nanorods exhibiting strong quantum confinement even at relatively large rod sizes, a near reciprocal relation between the 2ASE threshold and the 2PA action cross-section (σ(2)η) (where η is the quantum yield) was found and validated over a wide volume range for II-VI semiconductor nanostructures. Ultrafast optical spectroscopy verified that while the Auger processes in these heterostructures are indeed suppressed, ASE in these samples could also be strongly affected by a fast hole-trapping process to the NR surface states. Lastly, to exemplify the potential of these seeded CdSe/CdS nanodot/nanorod heterostructures as a viable gain media for achieving two-photon lasing, a highly photostable microsphere laser with an ultralow pump threshold is showcased.

Specialist Trainees on Rotation Cannot Replace Dedicated Consultant Clinicians for Antimicrobial Stewardship of Specialty Disciplines

ABSTRACT: Our prospective-audit-and-feedback antimicrobial stewardship (AS) program for hematology and oncology inpatients was switched from one led by dedicated clinicians to a rotating team of infectious diseases trainees in order to provide learning opportunities and attempt a "de-escalation" of specialist input towards a more protocol-driven implementation. However, process indicators including number of and compliance to recommendations fell significantly during the year, with accompanying increases in broad-spectrum antibiotic prescription. The trends were reversed only upon reverting to the original setup. Dedicated clinicians play a crucial role in AS programs involving immunocompromised patients. Structured training and adequate succession/contingency planning is critical for sustainability.

Drug-induced Dermatomyositis After Zoledronic Acid

A 57-year-old woman with prior exposure to bisphosphonates developed myalgia, proximal muscle weakness and lichenoid rash over the upper extremities and face 3 days after infusion of zoledronic acid for the management of osteoporosis. The diagnosis of dermatomyositis was made on the basis of clinical, laboratory and histological findings. This is the first report of drug-induced dermatomyositis secondary to zoledronic acid.

Using an Integrated COC Index and Multilevel Measurements to Verify the Care Outcome of Patients with Multiple Chronic Conditions

ABSTRACT: BACKGROUND: The increasing prevalence of multiple chronic conditions has accentuated the importance of coordinating and integrating health care services. Patients with better continuity of care (COC) have a lower utilization rate of emergency department (ED) services, lower hospitalization and better care outcomes. Previous COC studies have focused on the care outcome of patients with a single chronic condition or that of physician-patient relationships; few studies have investigated the care outcome of patients with multiple chronic conditions. Using multi-chronic patients as subjects, this study proposes an integrated continuity of care (ICOC) index to verify the association between COC and care outcomes for two scopes of chronic conditions, at physician and medical facility levels. METHODS: This study used a dataset of 280,840 subjects, obtained from the Longitudinal Health Insurance Database (LHID 2005), compiled by the National Health Research Institutes, of the National Health Insurance Bureau of Taiwan. Principal Component Analysis (PCA) was used to integrate the indices of density, dispersion and sequence into ICOC to measure COC outcomes - the utilization rate of ED services and hospitalization. A Generalized Estimating Equations model was used to verify the care outcomes. RESULTS: We discovered that the higher the COC at medical facility level, the lower the utilization rate of ED services and hospitalization for patients; by contrast, the higher the COC at physician level, the higher the utilization rate of ED services (odds ratio > 1; Exp(beta) = 2.116) and hospitalization (odds ratio > 1; Exp(beta) = 1.688). When only those patients with major chronic conditions with the highest number of medical visits were considered, it was found that the higher the COC at both medical facility and physician levels, the lower the utilization rate of ED services and hospitalization. CONCLUSIONS: The study shows that ICOC is more stable than single indices and it can be widely used to measure the care outcomes of different chronic conditions to accumulate empirical evidence. Concentrated care of multi-chronic patients by a single physician often results in unsatisfactory care outcomes. This highlights the need for referral mechanisms and integration of specialties inside or outside medical facilities, in order to optimize patient-centered care.

The Relationship Between Gallbladder Status and Recurrent Biliary Complications in Patients with Choledocholithiasis Following Endoscopic Treatment

Endoscopic methods are currently the treatment of choice for patients with common bile duct (CBD) stones, but subsequent management of the intact gallbladder for patients following endoscopic treatment is still controversial. The primary aim of this study was to discover the association between gallbladder status and recurrent biliary complications for patients with CBD stones after endoscopic treatment. Additionally, we also sought to determine risk factors for recurrent biliary complications in these patients.

Micromechanical Modeling of R-curve Behaviors in Human Cortical Bone

The risk of bone fracture increases with age because of a variety of factors that include, among others, decreasing bone quantity and quality due to increasing porosity and crack density with age. Experimental evidence has indicated that changes in bone microstructure and trace mineralization with age can result in different crack-tip strain field and fracture response, leading to different fracture mechanisms and R-curve behaviors. In this paper, a micromechanical modeling approach is developed to predict the R-curve response of bone tissue by delineating fracture mechanisms that lead to microdamage and ligament bridging by incorporating the influence of increasing porosity and crack density with age. The effects of age on fracture of human femur cortical bone due to porosity (bone quantity) and bone quality (crack density) with age are then examined via the micromechanical model.

Long-term Trends in the Epidemiology of Human Enteropathogens in Malaysia

Designing and Implementing a System for Tracking Functional Status After Stroke: A Feasibility Study

OBJECTIVE: To determine the feasibility of tracking stroke patients' functional outcomes in an integrated health system across a care continuum using the computer version of the Activity Measure of Post-Acute Care (AM-PAC). SETTING: A large integrated health care system in northern California. PARTICIPANTS: A total of 222 stroke patients (aged ≥18 years) who were hospitalized after an acute cerebrovascular accident. METHODS: An AM-PAC assessment was made at discharge from sites of care, including acute hospital, inpatient rehabilitation hospital, skilled nursing facility, home during home care, and outpatient settings. Assessments also were completed in the patient's home at 6 months. Data from the AM-PAC program were integrated with the health care system's databases. MAIN OUTCOME MEASUREMENTS: (1) AM-PAC administration time at the various sites of care; (2) assessment of a floor or a ceiling effect; and (3) administrative burden of tracking participants. RESULTS: AM-PAC assessment sessions averaged 7.9 minutes for data acquisition in 3 domains: Basic Mobility, Activities of Daily Living, and Applied Cognition. Participants answered, on average, 27 AM-PAC questions per session. A small ceiling effect was observed at 6 months, and there was a larger ceiling effect when the instrument was administered in an institution, ie, when the AM-PAC institutional item bank was used rather than the community item bank. It was feasible to track patients and to assess their function using the AM-PAC instrument from institutional to community settings. Implementation of the AM-PAC in clinical environments, and the success of the project, were influenced by instrumental, technological, operational, resource, and cultural factors. CONCLUSIONS: This study demonstrates the feasibility of implementing a single functional outcome instrument in clinical and community settings to measure rehabilitation functional outcomes of stroke patients. Integrating the AM-PAC measurement system into clinical workflows and the electronic medical record could provide assistance to clinicians for medical decision making, functional prognostication, and discharge planning.

Increased Risk of Advanced Neoplasms Among Asymptomatic Siblings of Patients with Colorectal Cancer

BACKGROUND & AIMS:: Colorectal cancer (CRC) is the second-most common cancer in Hong Kong. Relatives of patients with CRC have an increased risk of colorectal neoplasm. We assessed the prevalence of advanced neoplasms among asymptomatic siblings of patients with CRC. METHODS:: Patients with CRC were identified from the Prince of Wales Hospital CRC Surgery Registry from 2001 to 2011. Colonoscopies were performed for 374 siblings of patients (age 52.6±7.4 y) and 374 age- and sex-matched siblings of healthy subjects who had normal colonoscopies and did not have a family history of CRC (controls, 52.7± 7.4 y). We identified individuals with advanced neoplasms (defined as cancers or adenomas of at least 10 mm in diameter, high-grade dysplasia, villous or tubulovillous characteristics) RESULTS:: The prevalence of advanced neoplasms was 7.5% among siblings of patients and 2.9% among controls (matched odds ratio [mOR], 3.07; 95% confidence interval [CI], 1.5-6.3; P =.002). The prevalence of adenomas larger than 10 mm was higher among siblings of patients than in controls (5.9% vs 2.1%; mOR, 3.34; 95% CI, 1.45-7.66; P =.004), as was the presence of colorectal adenomas (31.0% vs 18.2%; mOR, 2.19; 95% CI, 1.52-3.17; P <.001). Six cancers were detected among siblings of patients; no cancers were detected in controls. The prevalence of advanced neoplasms among siblings of patients was higher when their index case was female (mOR, 4.95; 95% CI, 1.81-13.55) and had distally located CRC (mOR, 3.10; 95% CI, 1.34-7.14). CONCLUSIONS:: In Hong Kong, siblings of patients with CRC have a higher prevalence of advanced neoplasms, including CRC, than siblings of healthy individuals. Screening is indicated in this high-risk population. ClinicalTrials.gov number: NCT00164944.

Impact of Stylet Use in a Simulated Difficult Airway Model

BACKGROUND: Stylet use during endotracheal intubation (ETI) is variable across medical specialty and geographic location; however, few objective data exist regarding the impact of stylet use on ETI performance. OBJECTIVE: We evaluated the impact of stylet use on the time required to perform ETI in cases of simulated difficult airways in novice and experienced providers. METHODS: We performed a prospective, randomized observational study of experienced (attending anesthesiologists and emergency physicians) vs inexperienced airway providers (emergency medical technician, paramedic and medical students) comparing the use of stylet vs no stylet in random order using a simulated difficult airway. The primary outcome was attempt time for each of 6 attempts defined as entry of the laryngoscope in the mouth until successfully passing the endotracheal tube past the vocal cords. We analyzed the data using descriptive statistics including means with SDs and t tests. We used generalized estimating equations to evaluate potential changes in the attempt time over multiple attempts. RESULTS: There were 23 providers per group. The mean (SD) inexperienced attempt time in seconds was 25.88 (28.46) and 10.50 (5.47) for experienced providers (P < .0001). Stylet use did not alter attempt time for either group. When adjusting for stylet use, the attempt time did not change over repeated intubations (P = .541). When adjusting for experience status, inexperienced intubators had shorter attempt times with each successive trial, whereas experienced intubators attempt times remained constant (P < .001). CONCLUSION: Stylet use does not improve attempt time in a simulated difficult airway model for either inexperienced or experienced intubators.

Haploinsufficiency of the Paternal-effect Gene Dnmt3L Results in Transient DNA Hypomethylation in Progenitor Cells of the Male Germline

STUDY QUESTION: How does haploinsufficiency of the paternal-effect gene Dnmt3L affect DNA methylation establishment and stability in the male germline? SUMMARY ANSWER: Reduced expression of DNMT3L in male germ cells, associated with haploinsufficiency of the paternal-effect gene Dnmt3L, results in abnormal hypomethylation of prenatal germline progenitor cells. WHAT IS KNOWN ALREADY: The DNA methyltransferase regulator Dnmt3-Like (Dnmt3L) is a paternal-effect gene required for DNA methylation acquisition in male germline stem cells and their precursors. In males, DNMT3L deficiency causes meiotic abnormalities and infertility. While Dnmt3L heterozygous males are fertile, they have abnormalities in X chromosome compaction and postmeiotic gene expression and sire offspring with sex chromosome aneuploidy. It has been proposed that the paternal effects of Dnmt3L haploinsufficiency are due to epigenetic defects in early male germ cells. DNA methylation is an essential epigenetic modification essential for normal germ cell development. Since patterns of DNA methylation across the genome are initially acquired in prenatal male germ cells, perturbations in methylation could contribute to the epigenetic basis of the paternal effects in Dnmt3L(+/-) males. STUDY DESIGN, SIZE, DURATION: This is a cross-sectional study of DNA methylation in Dnmt3L(+/+) versus Dnmt3L(+/-) male germ cells collected from mice at 16.5 days post-coitum (dpc), Day 6 and Day 70 (n = 3 per genotype, each n represents a pool of 2-20 animals). Additionally, DNA methylation was compared in enriched populations of spermatogonial stem cells (SSC)/progenitor cells from Dnmt3L(+/+) and Dnmt3L(+/-) males following ∼2 months in culture. MATERIALS, SETTING, METHODS: DNA methylation at intergenic loci along chromosomes 9 and X was examined by quantitative analysis of DNA methylation by real-time polymerase chain reaction at the time of initial acquisition of epigenetic patterns in the prenatal male germline (16.5 dpc) and compared with patterns in early post-natal spermatogonia (Day 6) and in spermatozoa in mice. DNA methylation status at CpG-rich sites across the genome was assessed in spermatogonial precursors from Day 4 male mice using restriction landmark genomic scanning. MAIN RESULTS AND THE ROLE OF CHANCE: At 16.5 dpc, 42% of intergenic loci examined along chromosome 9 and 10% of those along chromosome X were hypomethylated in Dnmt3L heterozygotes. By Day 6 and in spermatozoa, germ cell DNA methylation was similar in heterozygous and wild-type mice. DNA methylation stability of acquired patterns in wild-type and Dnmt3L(+/-) SSC/progenitor cell culture was analyzed at numerous loci across the genome in cells cultured in vitro and collected at passages 6-28. While the methylation of most loci was stable in culture over time, differences at ∼1% of sites were found between Dnmt3L(+/-) and Dnmt3L(+/+) cultures. LIMITATIONS, REASONS FOR CAUTION: Evaluation of DNA methylation in SSCs can only be performed after a period of culture limiting the investigation to changes observed during culture when compared with DNA methylation differences between genotypes that could be present at the beginning of culture establishment. WIDER IMPLICATIONS OF THE FINDINGS: The DNA methylation defects described here in prenatal male germline progenitor cells and SSC culture are the earliest epigenetic perturbations yet identified for a mammalian paternal-effect gene and may influence downstream epigenetic events in germ cells at later stages of development. Together, the results provide evidence of a 'window' of susceptibility in prenatal male germ cell precursors for the induction of epimutations due to genetic perturbations and, potentially, in utero environmental exposures. STUDY FUNDING/COMPETING INTEREST(S): Canadian Institutes of Health Research (CIHR) provided funding for J.M.T. (MOP229913) and M.C.N. (MOP86532). The authors have no conflicts of interest to declare.

Biomedical Text Mining and Its Applications in Cancer Research

Cancer is a malignant disease that has caused millions of human deaths. Its study has a long history of well over 100years. There have been an enormous number of publications on cancer research. This integrated but unstructured biomedical text is of great value for cancer diagnostics, treatment, and prevention. The immense body and rapid growth of biomedical text on cancer has led to the appearance of a large number of text mining techniques aimed at extracting novel knowledge from scientific text. Biomedical text mining on cancer research is computationally automatic and high-throughput in nature. However, it is error-prone due to the complexity of natural language processing. In this review, we introduce the basic concepts underlying text mining and examine some frequently used algorithms, tools, and data sets, as well as assessing how much these algorithms have been utilized. We then discuss the current state-of-the-art text mining applications in cancer research and we also provide some resources for cancer text mining. With the development of systems biology, researchers tend to understand complex biomedical systems from a systems biology viewpoint. Thus, the full utilization of text mining to facilitate cancer systems biology research is fast becoming a major concern. To address this issue, we describe the general workflow of text mining in cancer systems biology and each phase of the workflow. We hope that this review can (i) provide a useful overview of the current work of this field; (ii) help researchers to choose text mining tools and datasets; and (iii) highlight how to apply text mining to assist cancer systems biology research.

The Magnitude of Global Marine Species Diversity

BACKGROUND: The question of how many marine species exist is important because it provides a metric for how much we do and do not know about life in the oceans. We have compiled the first register of the marine species of the world and used this baseline to estimate how many more species, partitioned among all major eukaryotic groups, may be discovered. RESULTS: There are ∼226,000 eukaryotic marine species described. More species were described in the past decade (∼20,000) than in any previous one. The number of authors describing new species has been increasing at a faster rate than the number of new species described in the past six decades. We report that there are ∼170,000 synonyms, that 58,000-72,000 species are collected but not yet described, and that 482,000-741,000 more species have yet to be sampled. Molecular methods may add tens of thousands of cryptic species. Thus, there may be 0.7-1.0 million marine species. Past rates of description of new species indicate there may be 0.5 ± 0.2 million marine species. On average 37% (median 31%) of species in over 100 recent field studies around the world might be new to science. CONCLUSIONS: Currently, between one-third and two-thirds of marine species may be undescribed, and previous estimates of there being well over one million marine species appear highly unlikely. More species than ever before are being described annually by an increasing number of authors. If the current trend continues, most species will be discovered this century.

Cell Phone Use and Traffic Crash Risk: a Culpability Analysis

BACKGROUND: The use of a cell phone or communication device while driving is illegal in many jurisdictions, yet evidence evaluating the crash risk associated with cell phone use in naturalistic settings is limited. This article aims to determine whether cell phone use while driving increases motor vehicle crash culpability.Method Drivers involved in crashes where police reported cell phone use (n = 312) and propensity matched drivers (age, sex, suspect alcohol/drug impairment, crash type, date, time of day, geographical location) without cell phone use (n = 936) were drawn from Insurance Corporation of British Columbia Traffic Accident System data. A standardized scoring tool, modified to account for Canadian driving conditions, was used to determine crash culpability from police reports on all drivers from the crashes. The association between crash culpability and cell phone use was determined, with additional subgroup analyses based on crash severity, driver characteristics and type of licence. RESULTS: A comparison of crashes with vs without cell phones revealed an odds ratio of 1.70 (95% confidence interval 1.22-2.36; P = 0.002). This association was consistent after adjustment for matching variables and other covariates. Subgroup analyses demonstrated an association for male drivers, unimpaired drivers, injured and non-injured drivers, and for drivers aged between 26 and 65 years. CONCLUSIONS: Crash culpability was found to be significantly associated with cell phone use by drivers, increasing the odds of a culpable crash by 70% compared with drivers who did not use a cell phone. This increased risk was particularly high for middle-aged drivers.

The Evolution and Refinement of Vasoepididymostomy Techniques

Obstructive azoospermia secondary to epididymal obstruction can be corrected by microsurgical reconstruction with vasoepididymostomy (VE). Although alternative management such as epididymal or testicular sperm aspiration in conjunction with intracytoplasmic sperm injection is feasible, various studies have established the superior cost-effectiveness of VE as a treatment of choice. Microsurgical VE is considered one of the most technically challenging microsurgeries. Its success rate is highly dependent on the skills and experience of the surgeons. Various techniques have been described in the literature for VE. We have pioneered a technique known as longitudinal intussusception VE (LIVE) in which the epididymal tubule is opened longitudinally to obtain a larger opening to allow its tubular content to pass through the anastomosis. Our preliminary data demonstrated a patency rate of over 90%. This technique has been widely referenced in the recent literature including robotic-assisted microsurgery. The history of the development of different VE approaches, the preoperative evaluation along with the techniques of various VE will be described in this article.Asian Journal of Andrology advance online publication, 19 November 2012; doi:10.1038/aja.2012.80.

Text Mining in the Biocuration Workflow: Applications for Literature Curation at WormBase, DictyBase and TAIR

WormBase, dictyBase and The Arabidopsis Information Resource (TAIR) are model organism databases containing information about Caenorhabditis elegans and other nematodes, the social amoeba Dictyostelium discoideum and related Dictyostelids and the flowering plant Arabidopsis thaliana, respectively. Each database curates multiple data types from the primary research literature. In this article, we describe the curation workflow at WormBase, with particular emphasis on our use of text-mining tools (BioCreative 2012, Workshop Track II). We then describe the application of a specific component of that workflow, Textpresso for Cellular Component Curation (CCC), to Gene Ontology (GO) curation at dictyBase and TAIR (BioCreative 2012, Workshop Track III). We find that, with organism-specific modifications, Textpresso can be used by dictyBase and TAIR to annotate gene productions to GO's Cellular Component (CC) ontology.

Clinical Practice Implications of Immunizations After Pediatric Bone Marrow Transplant: A Literature Review

The number of pediatric bone marrow transplants is increasing for malignant and nonmalignant diseases. The number of survivors is also increasing, and their long-term health and protection from infection is increasingly important. To prevent infections, it is standard practice to re-immunize pediatric patients after bone marrow transplant (BMT) using the Centers for Disease Control and Prevention immunization guidelines; however, surveys in the United States and other parts of the world indicate that many BMT patients do not receive all the recommended immunizations. A literature review was conducted to identify research based on evidence for immunization following BMT and to recognize barriers to the process. Also, the immunization clinical guidelines from 2000 and 2011 for patients following BMT were compared and an updated clinical protocol and immunization schedule was developed to reflect the current evidence, encourage a change in practice, and discourage fragmented care.

Hyaluronic Acid-dependent Protection Against Alkali-burned Human Corneal Cells

Hyaluronic acid (HA) is a high-molecular weight glycosaminoglycan and extracellular matrix component that promotes cell proliferation. This study aimed to evaluate the effects of HA on alkali-injured human corneal epithelial cells in vitro, and to elucidate the mechanisms by which HA mediates corneal cell protection. A human corneal epithelial cell line (HCE-2) was treated with sodium hydroxide (NaOH) before incubation with low-molecular weight HA (LMW-HA, 127 kDa) or high-molecular weight HA (HMW-HA, 1525 kDa). A global proteomic analysis was then performed. Our data indicated that HA treatment protects corneal epithelial cells from alkali injury, and that the molecular weight of HA is a crucial factor in determining its effects. Only HMW-HA reduced NaOH-induced cytotoxic effects in corneal cells significantly and increased their migratory and wound healing ability. Results from 2D-DIGE and MALDI-TOF/TOF MS analyses indicated that HMW-HA modulates biosynthetic pathways, cell migration, cell outgrowth, and protein degradation to stimulate wound healing and prevent cell death. To our knowledge, our study is the first to report the possible mechanisms by which HMW-HA promotes repair in alkali-injured human corneal epithelial cells.

Life-Course Socioeconomic Status and Obesity Among Older Singaporean Chinese Men and Women

Objectives.To elucidate the association between life-course socioeconomic status (SES) and obesity among older (aged 60 and older) Singaporean Chinese men and women. METHODS: Data from the Social Isolation, Health and Lifestyles Survey (single-stage stratified random sampling design) was utilized. Obesity (body mass index >27.4kg/m(2)) was assessed for 1,530 men and 2,036 women. Childhood (family financial status while growing up), adult (education), and older adult (housing type) SES indicators were used to define the accumulation of risk (cumulative socioeconomic disadvantage), social mobility (8 trajectories using the 3 SES indicators), and sensitive period (independent effect of each SES indicator) conceptual models. Association between the 3 life-course SES conceptual models and obesity was assessed using logistic regression analysis. RESULTS: Among women and men, low childhood SES lowered the odds of obesity. Low adult SES increased the odds of obesity only among women. There was no association between cumulative socioeconomic disadvantage and obesity. Women experiencing upward social mobility had lower odds of obesity relative to both those experiencing low SES and high SES through the life-course.Discussion.Association of the life-course SES conceptual models with obesity among older Singaporeans is different from that reported among younger Western populations, suggesting the association to be context specific. The different conceptual models complement each other.

Mucosal Melanoma of the Head and Neck: 32-year Experience in a Tertiary Referral Hospital

INTRODUCTION: Primary mucosal melanomas of the head and neck (HNMM), albeit being rare, are rapidly lethal. Here we report the experience of patients with HNMM treated in our institution over a 32-year period. OBJECTIVES/HYPOTHESIS: We aim to review our experience in managing HNMM patients over a 32-year period. STUDY DESIGN: Retrospective study. METHODS: Thirty-five patients diagnosed with HNMM from 1978 to 2009 were retrospectively reviewed, with an emphasis on predictors on survival outcome. RESULTS: Twenty-four patients received curative resection, 6 of them followed by adjuvant radiotherapy. Neck dissections were performed in 8 patients. Four patients received radiotherapy as primary treatment. Seven patients were treated conservatively. The overall mean and median survivals were 50 and 26 months, respectively. The median survival of stage I, II, and III diseases in our group of patients were 39, 10, and 16 months, respectively. The 1-year and 5-year overall survival rates were 65.7% and 22.9%, respectively. Age above 60 (p = 0.007), nodal involvement (p = 0.047;) and stage at presentation (p = 0.046) were shown to be associated with worse overall survival. Sites of tumour did not seem to impact on survival. On multivariate analysis, only age (below or above 60) was found to be statistically significant [RR 4.79 (1.65-13.9), p = 0.004]. CONCLUSIONS: Oral cavity melanomas are more likely to have nodal involvement at presentation. Prognosis of HNMM remains grave. Current evidence still supports surgery as the best chance of cure. Role of adjuvant radiotherapy is controversial and does not appear to improve overall survival. Similarly, role of neck dissection is ill-defined. Laryngoscope, 2012.

Investigation of the BRAF V600E Mutation by Pyrosequencing in Lymphoproliferative Disorders

The presence of the BRAF c.1799T>A V600E mutation was recently described in cases of hairy cell leukemia (HCL) but not in other common lymphomas. However, many uncommon subtypes of lymphoma have not been studied. We designed a BRAF pyrosequencing assay specific for the V600E mutation, which has a sensitivity of 5% and is applicable to paraffin-embedded tissue. DNA was sequenced in 9 cases of HCL; 6 cases of variant HCL; 10 cases each of nodal marginal zone lymphoma (NMZL), extranodal marginal zone lymphoma (ENMZL), posttransplantation lymphoproliferative disorder (PTLD), and large granular lymphocyte (LGL) proliferations; 11 cases of peripheral T-cell lymphoma (PTCL); and 12 cases of anaplastic large cell lymphoma (ALCL). All (100%) cases of HCL were positive for BRAF mutations. No mutations were identified in variant HCL, NMZL, ENMZL, PTLD, PTCL, ALCL, or LGL proliferations. Among lymphoproliferative disorders, BRAF mutations are restricted to HCL.

Paronychia Induced by the Epidermal Growth Factor Receptor Inhibitor Cetuximab

While the development of epidermal growth factor receptor inhibitors has been hailed as a remarkable triumph in the field of oncology, it has inherited with it a host of cutaneous side-effects that have been increasingly observed in a substantial number of patients in the recent years. One cutaneous manifestation that may inflict significant pain and affect activities of daily living among some of the patients receiving epidermal growth factor receptor inhibitors is paronychia. A case of paronychia associated with the use of cetuximab in the management of KRAS wild-type midrectal adenocarcinoma along with its management has been described.

Need for Rigor in Design, Reporting, and Interpretation of Transcriptomic Biomarker Studies

Calcium-43 NMR Studies of Polymorphic Transition of Calcite to Aragonite

Phase transformation between calcite and aragonite is an important issue in biomineralization. To shed more light on the mechanism of this process at the molecular level, we employ solid-state (43)Ca NMR to study the phase transformation from calcite to aragonite as regulated by magnesium ions, with (43)Ca enrichment at a level of 6%. Using the gas diffusion approach, the phase of Mg-calcite is formed initially and the system subsequently transforms to aragonite as the reaction time proceeds. Our (43)Ca solid-state NMR data support the dissolution-recrystallization mechanism for the calcite to aragonite transition. We find that the (43)Ca NMR parameters of Mg-calcite are very similar to those of pure calcite. Under the high-resolution condition provided by magic-angle spinning at 4 kHz, we can monitor the variation of the (43)Ca NMR parameters of the aragonite signals for the samples obtained at different reaction times. Our data suggest that in the presence of a significant amount of Mg(2+) ions, aragonite is the most stable polymorph of calcium carbonate. The initial precipitated crystallites of aragonite have spine-like morphology, for which the (43)Ca spin-lattice relaxation data indicate that the ions in the lattice have considerable motional dynamics. As the crystallinity of aragonite improves further, the (43)Ca T(1) parameter of the aragonite phase changes considerably and becomes very similar to that obtained for pure aragonite. For the first time, the difference in crystal morphologies and crystallinity of the aragonite phase has been traced down to the subtle difference in the motional dynamics at the molecular level.

Commentary: Risk Factors for Gastrointestinal Bleeding in NSAID Users - Authors' Reply

A Meta-Analysis of Acculturation/Enculturation and Mental Health

This meta-analytic study examined the relationship among the constructs of acculturation, enculturation, and acculturation strategies (i.e., integration, assimilation, separation, marginalization), and mental health. Data from 325 studies (163 journal articles and 162 dissertation studies) were analyzed using a random-effects model, across a broad spectrum of negative mental health (NM: depression, anxiety, psychological distress, and negative affect) and positive mental health (PM: self-esteem, satisfaction with life, and positive affect). Overall, acculturation was favorably associated with both NM (negatively) and PM (positively), whereas enculturation was favorably related only to PM (positively). In fact, enculturation was positively related to anxiety. The specifics of these relations were further examined using the following moderators: (a) researchers' operationalization of acculturation/enculturation (i.e., linearity, dimensionality); (b) contextual influences (i.e., when and where the study was conducted); and (c) sample characteristics (i.e., voluntariness of residency, race, gender, age). Overall, bilinear measures of acculturation indicated a positive association with PM, while unilinear measures did not. External acculturation (e.g., language, behaviors) and internal enculturation (e.g., identity) were most favorably related to mental health. The place of study had differential effects on the relation of enculturation and NM. Acculturation appeared to be especially important to Asian Americans, whereas enculturation was to African Americans. Differential effects of age suggested the need to consider life-span development of needs and social roles in relation to acculturation and enculturation. Both correlational analyses and mean comparisons affirmed that integration was the most favorable acculturation strategy to mental health. Implications for research, practice, and theory are discussed. (PsycINFO Database Record (c) 2012 APA, all rights reserved).

On the Time Evolution of the Cloaking Effect of a Metamaterial Slab

We investigated the time evolution of the cloaking behavior of a small particle placed in front of a meta-material slab with ε=μ=-1+iδ. We found that the dipole excitation would be suppressed in the long time limit. While on the way to being cloaked, the excitation will exhibit oscillatory behavior as the result of the interference between particle-slab resonances and high density-of-states surface modes.

Targeting Gonadotropin-releasing Hormone Receptor Inhibits the Early Step of Ovarian Cancer Metastasis by Modulating Tumor-mesothelial Adhesion

Ovarian cancer has a clear predilection to metastasize to the peritoneum, which represents one of the most important prognostic factors of poor clinical outcome. Gonadotropin-releasing hormone (GnRH) receptor is significantly overexpressed during the malignant progression of human ovarian cancer. Here, using lentiviral-based small interfering RNA (siRNA) technology to downregulate GnRH receptor in metastatic ovarian cancer cells, we show that GnRH receptor is an important mediator of ovarian cancer peritoneal metastasis. GnRH receptor downregulation dramatically attenuated their adhesion to the peritoneal mesothelium. By inhibiting the expression of GnRH receptor, we showed decreased expression of α2β1 and α5β1 integrin and adhesion to specific extracellular matrix (ECM) proteins. This was also associated with a reduction of P-cadherin. Furthermore, adhesion of ovarian cancer cells to different ECMs and the mesothelium were abrogated in response to β1 integrin and P-cadherin reduction, confirming that the effects were β1 integrin- and P-cadherin-specific. Using a mouse model of human ovarian cancer metastasis, we found that the inhibition of GnRH receptor, β1 integrin, and P-cadherin significantly attenuated tumor growth, ascites formation, and the number of metastatic implants. These results define a new role for GnRH receptor in early metastasis and offer the possibility of novel therapeutic targets.Molecular Therapy (2012); doi:10.1038/mt.2012.187.

A Multilevel Approach for Assessing the Variability of Hepatitis C Prevalence in Injection Drug Users by Their Gathering Places

OBJECTIVES: The aim of this study was to investigate the variation in hepatitis C virus (HCV) prevalence in injection drug users (IDUs) by their gathering places, using a multilevel approach. METHODS: IDUs recruited from their gathering places were invited to respond to a questionnaire on demographics, drug use history, injection behaviors, and methadone treatment. Dried blood spots were collected for HCV antibody testing by ELISA. Factors associated with the anti-HCV test result were explored by linear logistic regression, followed by the evaluation of heterogeneity between gathering places by multilevel analysis. RESULTS: A total of 622 respondents from 19 gathering places in Hong Kong, recruited between August and September 2011, were evaluated. Anti-HCV seroprevalence was 81.7% (95% confidence interval 78.6-84.7%), ranging from 67% to 100% by gathering place. HCV infection was associated with current practice of injection, needle-sharing, and midazolam injection. On multilevel analysis, there was a modest but significant variation in HCV antibody prevalence by gathering place, adjusted by midazolam injection (adjusted odds ratio (AOR) 3.91) and current injection (AOR 2.88) or injection over a long duration (AOR 3.17). CONCLUSIONS: There was heterogeneity in HCV antibody prevalence in IDUs by gathering place, while the influence of injection behaviors varied, suggesting interactivity between factors at the individual and group levels.

Targeting SHP2 Phosphatase in Myeloproliferative Neoplasms

A combined understanding of Shp2's structural requirement as well as defining the appropriate cellular population(s) for targeting Shp2 is likely to aid in the design of better Shp2 inhibitors in the future.

The Triterpenoid Cucurbitacin B Augments the Anti-proliferative Activity of Chemotherapy in Human Breast Cancer

Despite recent advances in therapy, breast cancer remains the second most common cause of death from malignancy in women. Chemotherapy plays a major role in breast cancer management, and combining chemotherapeutic agents with non-chemotherapeutic agents is of considerable clinical interest. Cucurbitacins are triterpenes compounds found in plants of the Cucurbitaceae family, reported to have anti-cancer and anti-inflamatory activities. Previously, we have shown antiproliferative activity of cucurbitacin B (CuB) in breast cancer, and we hypothesized that combining CuB with chemotherapeutic agents can augment their anti-tumor effect. Here, we show that a combination of CuB with either docetaxel (DOC) or gemcitabine (GEM) synergistically inhibited the proliferation of MDA-MB-231 breast cancer cells in vitro. This antiproliferative effect was accompanied by an increase in apoptosis rates. Furthermore, in vivo treatment of human breast cancer orthotopic xenografts in immunodeficient mice with CuB at either low (0.5 mg/kg) or high (1 mg/kg) doses in combination with either DOC (20 mg/kg) or GEM (12.5mg/kg) significantly reduced tumor volume as compared to monotherapy of each drug. Importantly, no significant toxicity was noted with low dose CuB in combination with either DOC or GEM. In conclusion, combination of CuB at a relatively low concentration with either of the chemotherapeutic agents, DOC or GEM, shows prominent antiproliferative activity against breast cancer cells without increased toxicity. This promising combination should be examined in therapeutic trials of breast cancer. © 2012 Wiley Periodicals, Inc.

The Correlations of LMX1A and Osteopontin Expression to the Clinicopathologic Stages in Pancreatic Adenocarcinoma

AIM:: To test the association of LMX1A and osteopontin (OPN) expression with histologic differentiation or pathologic stage in pancreatic ductal adenocarcinoma. METHODS:: Immunohistochemical analysis was performed to determine LMX1A and OPN expression in 100 surgical specimens obtained from Chinese patients with well-differentiated (n=15), moderately differentiated (n=65), and poorly differentiated (n=20) pancreatic ductal adenocarcinomas. RESULTS:: LMX1A and OPN immunoreactivities were undetectable in normal pancreatic glandular epithelia. Stronger immunostaining for LMX1A and OPN was associated with advanced nuclear grades of pancreatic ductal adenocarcinomas (70.7 and 87.1 for grade I, 109.8 and 118.3 for grade II, and 171.3 and 183.8 for grade III), and advanced TNM and American Joint Committee on Cancer stages of pancreatic ductal adenocarcinomas. CONCLUSIONS:: A higher expression of LMX1A and OPN is well correlated with histologic grade and pathologic stage of pancreatic ductal adenocarcinomas.

Retrospective Review of Positron Emission Tomography with Contrast-enhanced Computed Tomography in the Posttreatment Setting in Human Papillomavirus-associated Oropharyngeal Carcinoma

OBJECTIVE To determine the value of positron emission tomography (PET) with contrast-enhanced computed tomography (CT) in assessing the need for neck dissection by retrospectively reviewing the pathology reports of patients with human papillomavirus (HPV)-associated oropharyngeal squamous cell carcinoma (SCC). DESIGN Retrospective cohort study. SETTING Tertiary medical center. PATIENTS Seventy-seven patients with HPV-related SCC. MAIN OUTCOME MEASURES Seventy-seven consecutive patients with a diagnosis of HPV-related SCC who were treated with radiotherapy as the primary treatment between August 2007 and October 2010 were retrospectively evaluated for radiologic and pathologic rate of persistence of nodal metastasis after completion of definitive radiotherapy. Pretreatment and posttreatment imaging included contrast-enhanced CT and PET. Response to treatment was measured on CT, PET at standardized uptake value (SUV) thresholds of 2 and 2.5, and PET/CT by a neuroradiologist in a blinded fashion. Then, the pathology report of the patients who underwent neck dissections was reviewed for nodal status after resection and correlated with the imaging findings. RESULTS Of the 77 patients, 67 met the study criteria, with an average follow-up PET/CT scan at 90.5 days after completion of radiotherapy. Ten patients did not undergo follow-up PET/CT imaging. Twenty patients underwent neck dissections after completion of radiation therapy. Of these 20 patients, 4 had persistent tumor and 16 did not have viable tumor. Using the final pathology report to correlate with imaging responses, CT had a negative predictive value (NPV) of 85.7% (95% CI, 48.7%-97.4%), PET with SUV thresholds of 2 had an NPV of 91.7% (95% CI, 64.6%-98.5%), PET with a cutoff SUV of 2.5 had an NPV of 85.7% (95% CI, 60.1%-96.0%), PET/CT with an SUV of 2 had an NPV of 100% (95% CI, 59.8%-100.0%), and PET/CT with an SUV of 2.5 had an NPV of 85.7% (95% CI, 48.7%-97.4%). The 47 patients who did not undergo neck dissection had a median follow-up of 26 months without an isolated neck failure. Analysis of all 67 patients in the cohort revealed the following values: CT had an NPV of 95.7% (95% CI, 85.8%-98.8%), PET with an SUV of 2 had an NPV of 98.2% (95% CI, 90.4%-99.7%), PET with an SUV of 2.5 had an NPV of 95.0% (95% CI, 86.3%-98.3%), PET/CT with an SUV of 2 had an NPV of 100.0% (95% CI, 92.0%-100.0%), and PET/CT with an SUV of 2.5 had an NPV of 95.7% (95% CI, 85.8%-98.8%). CONCLUSIONS Positron emission tomography combined with contrast-enhanced CT has a better NPV than either imaging modality alone in patients with HPV-associated oropharyngeal SCC. Furthermore, PET/CT with an SUV threshold of 2 used in patients with HPV-related SCC offers an imaging modality with a high NPV that may obviate the need for unnecessary neck dissections.

Targeted Nanoparticle Delivery Overcomes Off-target Immunostimulatory Effects of Oligonucleotides and Improves Therapeutic Efficacy in Chronic Lymphocytic Leukemia

Several RNA targeted therapeutics including antisense oligonucleotides, small interfering RNAs and microRNAs constitute immunostimulatory CpG motifs as integral part of their design. The limited success with free anti-sense oligonucleotides in hematological malignancies in recent clinical trials has been attributed to the CpG motif mediated TLR induced pro-survival effects and inefficient target modulation in desired cells. In an attempt to diminish their off-target pro-survival and pro-inflammatory effects and specific delivery, as a proof of principle, we developed an antibody-targeted liposomal delivery strategy using a clinically relevant CD20 antibody (Rituximab) conjugated lipopolyplex nanoparticles (RIT-INPs) and Bcl-2 targeted anti-sense G3139 as archetypical antisense therapeutics. The adverse immunostimulatory responses were abrogated by selective B cell targeted delivery and early endosomal compartmentalization of G3139 encapsulated RIT-INPs, resulting in reduced NF-kB activation, robust Bcl-2 down-regulation and enhanced sensitivity to fludarabine induced cytotoxicity. Furthermore, significant in-vivo therapeutic efficacy was noted following RIT-INP-G3139 administration in a disseminated xenograft leukemia model. These results demonstrate that CD20 targeted delivery overcomes the immunostimulatory properties of CpG containing oligonucleotide therapeutics and improves efficient gene silencing and in vivo therapeutic efficacy for B-cell malignancies. The broader implications of similar approaches in overcoming immunostimulatory properties of RNA directed therapeutics in hematological malignancies are discussed.

Facile Synthesis and Characterization of Highly Fluorescent and Biocompatible N-acetyl-l-cysteine Capped CdTe/CdS/ZnS Core/shell/shell Quantum Dots in Aqueous Phase

The synthesis of water-soluble quantum dots (QDs) in aqueous phase has received much attention recently. To date various kinds of QDs such as CdTe, CdSe, CdTe/CdS and CdSe/ZnS have been synthesized by aqueous methods. However, generally poor-quality QDs (photoluminescent quantum yield (PLQY) lower than 30%) are obtained via this method and the 3-mercaptopropionic acid stabilizer is notorious for its toxicity and awful odor. Here we introduce a novel thiol ligand, N-acetyl-l-cysteine, as an ideal stabilizer that is successfully employed to synthesize high-quality CdTe/CdS/ZnS QDs via a simple aqueous phase. The core/shell/shell structures of the CdTe/CdS/ZnS QDs were verified by x-ray photoelectron spectroscopy, energy dispersive x-ray spectroscopy, x-ray powder diffraction and transmission electron microscopy. These QDs not only possess a high PLQY but also have excellent photostability and favorable biocompatibility, which is vital for many biological applications. This type of water-dispersed QD is a promising candidate for fluorescent probes in biological and medical fields.

Review of Vascularised Bone Tissue-engineering Strategies with a Focus on Co-culture Systems

Poor angiogenesis within tissue-engineered grafts has been identified as a main challenge limiting the clinical introduction of bone tissue-engineering (BTE) approaches for the repair of large bone defects. Thick BTE grafts often exhibit poor cellular viability particularly at the core, leading to graft failure and lack of integration with host tissues. Various BTE approaches have been explored for improving vascularisation in tissue-engineered constructs and are briefly discussed in this review. Recent investigations relating to co-culture systems of endothelial and osteoblast-like cells have shown evidence of BTE efficacy in increasing vascularization in thick constructs. This review provides an overview of key concepts related to bone formation and then focuses on the current state of engineered vascularized co-culture systems using bone repair as a model. It will also address key questions regarding the generation of clinically relevant vascularized bone constructs as well as potential directions and considerations for research with the objective of pursuing engineered co-culture systems in other disciplines of vascularized regenerative medicine. The final objective is to generate serious and functional long-lasting vessels for sustainable angiogenesis that will enable enhanced cellular survival within thick voluminous bone grafts, thereby aiding in bone formation and remodelling in the long term. However, more evidence about the quality of blood vessels formed and its associated functional improvement in bone formation as well as a mechanistic understanding of their interactions are necessary for designing better therapeutic strategies for translation to clinical settings. Copyright © 2012 John Wiley & Sons, Ltd.

Cell Viability and Angiogenic Potential of a Bioartificial Adipose Substitute

An implantable scaffold pre-seeded with cells needs to remain viable and encourage rapid angiogenesis in order to replace injured tissues, especially for tissue defect repairs. We created a bioartificial adipose graft composed of an electrospun 3D nanofibrous scaffold and fat tissue excised from New Zealand white rabbits. Cell viability and angiogenesis potential of the bioartificial substitute were examined during four weeks of culture in Dulbecco's Modified Eagle Medium by immunohistochemical staining with LIVE/DEAD® cell kit and PECAM-1 antibody, respectively. In addition, a Matrigel® assay was performed to examine the possibility of blood vessels sprouting from the bioartificial graft. Our results showed that cells within the graft were viable and vascular tubes were present at week 4, while cells in a fat tissue block were dead in vitro. In addition, capillaries were observed sprouting from the graft into the Matrigel, demonstrating its angiogenic potential. We expect that improved cell viability and angiogenesis in the bioartificial substitute, compared to intact autologous graft, could potentially contribute to its survival following implantation. Copyright © 2012 John Wiley & Sons, Ltd.

Characterization of Novel Akermanite:poly-ε-caprolactone Scaffolds for Human Adipose-derived Stem Cells Bone Tissue Engineering

In this study, three different akermanite:poly-ε-caprolactone (PCL) composite scaffolds (wt%: 75:25, 50:50, 25:75) were characterized in terms of structure, compression strength, degradation rate and in vitro biocompatibility to human adipose-derived stem cells (hASC). Pure ceramic scaffolds [CellCeram(TM) , custom-made, 40:60 wt%; β-tricalcium phosphate (β-TCP):hydroxyapatite (HA); and akermanite] and PCL scaffolds served as experimental controls. Compared to ceramic scaffolds, the authors hypothesized that optimal akermanite:PCL composites would have improved compression strength and comparable biocompatibility to hASC. Electron microscopy analysis revealed that PCL-containing scaffolds had the highest porosity but CellCeram(TM) had the greatest pore size. In general, compression strength in PCL-containing scaffolds was greater than in ceramic scaffolds. PCL-containing scaffolds were also more stable in culture than ceramic scaffolds. Nonetheless, mass losses after 21 days were observed in all scaffold types. Reduced hASC metabolic activity and increased cell detachment were observed after acute exposure to akermanite:PCL extracts (wt%: 75:25, 50:50). Among the PCL-containing scaffolds, hASC cultured for 21 days on akermanite:PCL (wt%: 75:25) discs displayed the highest viability, increased expression of osteogenic markers (alkaline phosphatase and osteocalcin) and lowest IL-6 expression. Together, the results indicate that akermanite:PCL composites may have appropriate mechanical and biocompatibility properties for use as bone tissue scaffolds. Copyright © 2012 John Wiley & Sons, Ltd.

A Truncated AdeS Kinase Protein Generated by ISAba1 Insertion Correlates with Tigecycline Resistance in Acinetobacter Baumannii

Over-expression of AdeABC efflux pump stimulated continuously by the mutated AdeRS two component system has been found to result in antimicrobial resistance, even tigecycline (TGC) resistance, in multidrug-resistant Acinetobacter baumannii (MRAB). Although the insertion sequence, ISAba1, contributes to one of the AdeRS mutations, the detail mechanism remains unclear. In the present study we collected 130 TGC-resistant isolates from 317 carbapenem resistant MRAB (MRAB-C) isolates, and 38 of them were characterized with ISAba1 insertion in the adeS gene. The relationship between the expression of AdeABC efflux pump and TGC resistant was verified indirectly by successfully reducing TGC resistance with NMP, an efflux pump inhibitor. Further analysis showed that the remaining gene following the ISAba1 insertion was still transcribed to generate a truncated AdeS protein by the Pout promoter on ISAba1 instead of frame shift or pre-termination. Through introducing a series of recombinant adeRS constructs into a adeRS knockout strain, we demonstrated the truncated AdeS protein was constitutively produced and stimulating the expression of AdeABC efflux pump via interaction with AdeR. Our findings suggest a mechanism of antimicrobial resistance induced by an aberrant cytoplasmic sensor derived from an insertion element.

Microcirculation Inflammation Associates With Outcome in Renal Transplant Patients With De Novo Donor-Specific Antibodies

In renal transplant patients with de novo donor-specific antibodies (dnDSA) we studied the value of microcirculation inflammation (MI; defined by the addition of glomerulitis (g) and peritubular capillaritis (ptc) scores) to assess long-term graft survival in a retrospective cohort study. Out of all transplant patients with standard immunological risk (n = 638), 79 (12.4%) developed dnDSA and 58/79 (73%) had an indication biopsy at or after dnDSA development. Based on the MI score on that indication biopsy patients were categorized, MI0 (n = 26), MI1 + 2 (n = 21) and MI ≥ 3 (n = 11). The MI groups did not differ significantly pretransplantation, whereas posttransplantation higher MI scores developed more anti-HLA class I + II DSA (p = 0.011), showed more TCMR (p < 0.001) and showed a trend to C4d-positive staining (p = 0.059). Four-year graft survival estimates from time of indication biopsy were MI0 96.1%, MI1 + 2 76.1% and MI ≥ 3 17.1%; resulting in a 24-fold increased risk of graft failure in the MI ≥ 3 compared to the MI0 group (p = 0.003; 95% CI [3.0-196.0]). When adjusted for C4d, MI ≥ 3 still had a 21-fold increased risk of graft failure (p = 0.005; 95% CI [2.5-180.0]), while C4d positivity on indication biopsy lost significance. In renal transplant patients with de novo DSA, microcirculation inflammation, defined by g + ptc, associates with graft survival.

Eu2+-Activated Sr8ZnSc(PO4)7: A Novel Near-Ultraviolet Converting Yellow-Emitting Phosphor for White Light-Emitting Diodes

The crystal structure of Eu2+-activated Sr8ZnSc(PO4)7:Eu2+ phosphor was refined and determined from XRD profiles by the Rietveld refinement method using a synchrotron light source. This phosphor crystallizes in the monoclinic structure with the I2/a space group. The SZSP:xEu2+ phosphors showed a broad yellow emission band centered at 511 and 571 nm depending on the concentration of Eu2+, and the composition-optimized concentration of Eu2+ in the Sr8ZnSc(PO4)7:Eu2+ phosphor was determined to be 2 mol%. The estimated crystal-field splitting and CIE chromaticity coordinates of Sr8ZnSc(PO4)7:xEu2+ (x = 0.001-0.05 mol) were 20181 ~ 20983 cm-1 and (0.3835, 0.5074) ~ (0.4221, 0.5012), respectively, and the emission band showed a redshift from 547 to 571 nm with increasing Eu2+ concentration. The non-radiative transitions between the Eu2+ ions in the Sr8ZnSc(PO4)7 host were attributable to dipole-dipole interactions, and the critical distance was approximately 19.8 Ã…. The combination of a 400 nm NUV chip with a blend of Sr8ZnSc(PO4)7:0.02Eu2+ and BAM:Eu2+ phosphors (light converters) gave high color rendering indices between 79.38 to 92.88, correlated color temperatures between 4325 K ~ 7937 K, and tuned CIE chromaticity coordinates in the range (0.381, 0.435) ~ (0.294, 0.310), respectively, depending on the SZSP:0.02Eu2+/BAM:Eu2+ weight ratio. These results suggest that the Sr8ZnSc(PO4)7:0.02Eu2+/BAM:Eu2+ phosphor blend has potential applications in white NUV LEDs.

Electrical Probing of Magnetic Phase Transition and Domain Wall Motion in Single-Crystalline Mn(5)Ge(3) Nanowire

In this letter, the magnetic phase transition and domain wall motion in a single-crystalline Mn(5)Ge(3) nanowire were investigated by temperature-dependent magneto-transport measurements. The ferromagnetic Mn(5)Ge(3) nanowire was fabricated by fully germaniding a single-crystalline Ge nanowire through the solid-state reaction with Mn contacts upon thermal annealing at 450 (o)C. Temperature-dependent 4-probe resistance measurements on the Mn(5)Ge(3) nanowire showed a clear slope change near 300 K accompanied by a magnetic phase transition from ferromagnetism to paramagnetism. The transition temperature was able to be controlled by both axial and radial magnetic fields as the external magnetic field helped maintain the magnetization aligned in the Mn(5)Ge(3) nanowire. Near the magnetic phase transition, the critical behavior in the 1D system was characterized by a power-law relation with a critical exponent of α = 0.07±0.01. Besides, another interesting feature was revealed as a cusp at about 67 K in the first-order derivative of the nanowire resistance, which was attributed to a possible magnetic transition between two non-collinear and collinear ferromagnetic states in the Mn5Ge3 lattice. Furthermore, temperature-dependent magneto-transport measurements demonstrated a hysteretic, symmetric and stepwise axial magnetoresistance of the Mn(5)Ge(3) nanowire. The interesting features of abrupt jumps indicated the presence of multiple domain walls in the Mn(5)Ge(3) nanowire and the annihilation of domain walls driven by the magnetic field. The Kurkijärvi model was used to describe the domain wall depinning as thermally assisted escape from a single energy barrier, and the fitting on the temperature dependent depinning magnetic fields yielded an energy barrier of 0.166 eV.

Psychometric Assessment of the Chinese Version of the Decisional Conflict Scale in Chinese Women Making Decision for Breast Cancer Surgery

OBJECTIVE: The decisional conflict scale (DCS) measures the perception of uncertainty in choosing options, factors contributing to decision conflict and effective decision making. This study examined the validity and reliability of the Chinese version of the DCS in Hong Kong Chinese women deciding breast cancer (BC) surgery. METHOD: A Chinese version of the 16-item DCS was administered to 471 women awaiting initial consultation for BC diagnosis. Confirmatory factor analysis (CFA) assessed the factor structure. Internal consistency, and convergent and discriminant validities of the factor structure were assessed. RESULTS: CFA revealed the original factor structure of the DCS showed poor fit to this sample. Exploratory factor analysis revealed an alternative three-factor structure, Informed and Values Clarity, Uncertainty and Effective Decision and Support, was optimal. Cronbach's alpha ranged from 0.51 to 0.87. Correlations between decision-making difficulties and satisfaction with medical consultation demonstrated acceptable convergent validity. Construct validity was supported by correlations between decision regret and psychological distress. Discriminant validity was supported by differentiation between delaying and non-delaying decision-makers. CONCLUSIONS: The three-factor DCS-14 is a valid and practical measure for assessing decisional conflict in deciding BC surgery. It shows good potential for use in assessing decision satisfaction for women diagnosed with BC.

Safety of Thromboprophylaxis After Oncologic Head and Neck Surgery. Study of 1018 Patients

BACKGROUND: Deep venous thrombosis (DVT) and pulmonary embolism (PE) are significantly reduced with appropriate use of thromboprophylaxis and scarcely evaluated in patients undergoing oncologic head and neck surgery (OHNS). METHODS: A retrospective study on 1018 patients who underwent oncologic head and neck surgery. The records of patients with venous thromboembolism (VTE) or postoperative bleeding were reviewed for the cancer grading, management, previous known coagulopathy, anticoagulation, and general demographics. RESULTS: Of a total of 1018 patients undergoing oncologic head and neck surgery, 450 patients had no chemoprophylaxis and 568 received it. The rate of a VTE event in our cohort was 0%. Twelve patients presented with hematoma or bleeding from the surgical site, 11 in the group that received chemoprophylaxis (p = .006). CONCLUSIONS: Our analysis shows no benefit from chemoprophylaxis in oncologic head and neck surgery patients, with no VTE events. Our analysis shows higher rates of morbid side effects from using chemoprophylaxis, and we therefore conclude that chemoprophylaxis should not be routinely used. © 2012 Wiley Periodicals, Inc. Head Neck, 2012.

Prognostic Significance of Circumferential Resection Margin Involvement Following Oesophagectomy for Cancer and the Predictive Role of Endoluminal Ultrasonography

Background:The optimum multimodal treatment for oesophageal cancer, and the prognostic significance of histopathological tumour involvement of the circumferential resection margin (CRM+) are uncertain. The aims of this study were to determine the prognostic significance of CRM+ after oesophagectomy and to identify endosonographic (endoluminal ultrasonography (EUS)) features that predict a threatened CRM+.Methods:Two hundred and sixty-nine consecutive patients underwent potentially curative oesophagectomy (103 surgery alone, 124 neoadjuvant chemotherapy (CS) and 42 chemoradiotherapy (CRTS)). Primary outcome measures were disease-free survival (DFS) and overall survival (OS).Results:CRM+ was reported in 98 (38.0%) of all, and in 90 (62.5%) of pT3 patients. Multivariate analysis of pathological factors revealed: lymphovascular invasion (HR 2.087, 95% CI 1.396-3.122, P<0.0001), CRM+ (HR 1.762, 95% CI 1.201-2.586, P=0.004) and lymph node metastasis count (HR 1.563, 95% CI 1.018-2.400, P=0.041) to be independently and significantly associated with DFS. Lymphovascular invasion (HR 2.160, 95% CI 1.432-3.259, P<0.001) and CRM+ (HR 1.514, 95% CI 1.000-2.292, P=0.050) were also independently and significantly associated with OS. Multivariate analysis revealed EUS T stage (T3 or T4, OR 24.313, 95% CI 7.438-79.476, P<0.0001) and use or not of CRTS (OR 0.116, 95% CI 0.035-0.382, P<0.0001) were independently and significantly associated with CRM+.Conclusion:A positive CRM was a better predictor of DFS and OS than standard pTNM stage.British Journal of Cancer advance online publication, 20 November 2012; doi:10.1038/bjc.2012.511 www.bjcancer.com.

In Vivo Directed Differentiation of Pluripotent Stem Cells for Skeletal Regeneration

Pluripotent cells represent a powerful tool for tissue regeneration, but their clinical utility is limited by their propensity to form teratomas. Little is known about their interaction with the surrounding niche following implantation and how this may be applied to promote survival and functional engraftment. In this study, we evaluated the ability of an osteogenic microniche consisting of a hydroxyapatite-coated, bone morphogenetic protein-2-releasing poly-l-lactic acid scaffold placed within the context of a macroenvironmental skeletal defect to guide in vivo differentiation of both embryonic and induced pluripotent stem cells. In this setting, we found de novo bone formation and participation by implanted cells in skeletal regeneration without the formation of a teratoma. This finding suggests that local cues from both the implanted scaffold/cell micro- and surrounding macroniche may act in concert to promote cellular survival and the in vivo acquisition of a terminal cell fate, thereby allowing for functional engraftment of pluripotent cells into regenerating tissue.

Identification of Small RNAs in Mycobacterium Smegmatis Using Heterologous Hfq

Gene regulation by small RNAs (sRNAs) has been extensively studied in various bacteria. However, the presence and roles of sRNAs in mycobacteria remain largely unclear. Immunoprecipitation of RNA chaperone Hfq to enrich for sRNAs is one of the effective methods to isolate sRNAs. However, the lack of an identified mycobacterial hfq restricts the feasibility of this approach. We developed a novel method that takes advantage of the conserved inherent sRNAs-binding capability of heterologous Hfq from Escherichia coli to enrich sRNAs from Mycobacterium smegmatis, a model organism for studying Mycobacterium tuberculosis. We validated 12 trans-encoded and 12 cis-encoded novel sRNAs in M. smegmatis. Many of these sRNAs are differentially expressed at exponential phase compared with stationary phase, suggesting that sRNAs are involved in the growth of mycobacteria. Intriguingly, five of the cis-encoded novel sRNAs target known transposases. Phylogenetic conservation analysis shows that these sRNAs are pathogenicity dependent. We believe that our findings will serve as an important reference for future analysis of sRNAs regulation in mycobacteria and will contribute significantly to the development of sRNAs prediction programs. Moreover, this novel method of using heterologous Hfq for sRNAs enrichment can be of general use for the discovery of bacterial sRNAs in which no endogenous Hfq is identified.

Interleukin 6-preconditioned Neural Stem Cells Reduce Ischaemic Injury in Stroke Mice

Transplantation of neural stem cells provides a promising therapy for stroke. Its efficacy, however, might be limited because of massive grafted-cell death after transplantation, and its insufficient capability for tissue repair. Interleukin 6 is a pro-inflammatory cytokine involved in the pathogenesis of various neurological disorders. Paradoxically, interleukin 6 promotes a pro-survival signalling pathway through activation of signal transducer and activator of transcription 3. In this study, we investigated whether cellular reprogramming of neural stem cells with interleukin 6 facilitates the effectiveness of cell transplantation therapy in ischaemic stroke. Neural stem cells harvested from the subventricular zone of foetal mice were preconditioned with interleukin 6 in vitro and transplanted into mouse brains 6 h or 7 days after transient middle cerebral artery occlusion. Interleukin 6 preconditioning protected the grafted neural stem cells from ischaemic reperfusion injury through signal transducer and activator of transcription 3-mediated upregulation of manganese superoxide dismutase, a primary mitochondrial antioxidant enzyme. In addition, interleukin 6 preconditioning induced secretion of vascular endothelial growth factor from the neural stem cells through activation of signal transducer and activator of transcription 3, resulting in promotion of angiogenesis in the ischaemic brain. Furthermore, transplantation of interleukin 6-preconditioned neural stem cells significantly attenuated infarct size and improved neurological performance compared with non-preconditioned neural stem cells. This interleukin 6-induced amelioration of ischaemic insults was abolished by transfecting the neural stem cells with signal transducer and activator of transcription 3 small interfering RNA before transplantation. These results indicate that preconditioning with interleukin 6, which reprograms neural stem cells to tolerate oxidative stress after ischaemic reperfusion injury and to induce angiogenesis through activation of signal transducer and activator of transcription 3, is a simple and beneficial approach for enhancing the effectiveness of cell transplantation therapy in ischaemic stroke.

Solitary Nodule on the Auricle

Online Disease Management of Diabetes: Engaging and Motivating Patients Online With Enhanced Resources-Diabetes (EMPOWER-D), a Randomized Controlled Trial

OBJECTIVE: To evaluate an online disease management system supporting patients with uncontrolled type 2 diabetes. MATERIALS AND METHODS: Engaging and Motivating Patients Online With Enhanced Resources for Diabetes was a 12-month parallel randomized controlled trial of 415 patients with type 2 diabetes with baseline glycosylated hemoglobin (A1C) values ≥7.5% from primary care sites sharing an electronic health record. The intervention included: (1) wirelessly uploaded home glucometer readings with graphical feedback; (2) comprehensive patient-specific diabetes summary status report; (3) nutrition and exercise logs; (4) insulin record; (5) online messaging with the patient's health team; (6) nurse care manager and dietitian providing advice and medication management; and (7) personalized text and video educational 'nuggets' dispensed electronically by the care team. A1C was the primary outcome variable. RESULTS: Compared with usual care (UC, n=189), patients in the intervention (INT, n=193) group had significantly reduced A1C at 6 months (-1.32% INT vs -0.66% UC; p<0.001). At 12 months, the differences were not significant (-1.14% INT vs -0.95% UC; p=0.133). In post hoc analysis, significantly more INT patients had improved diabetes control (>0.5% reduction in A1C) than UC patients at 12 months (69.9 (95% CI 63.2 to 76.5) vs 55.4 (95% CI 48.4 to 62.5); p=0.006). CONCLUSIONS: A nurse-led, multidisciplinary health team can manage a population of diabetic patients in an online disease management program. INT patients achieved greater decreases in A1C at 6 months than UC patients, but the differences were not sustained at 12 months. More INT than UC patients achieved improvement in A1C (>0.5% decrease).Trial registered in clinical trials.gov: #NCT00542204.

Characterization of Midface Fractures Incurred in Recent Wars

Facial injuries sustained by US military personnel during the wars in Iraq and Afghanistan have increased compared with past conflicts. Characterization of midface fractures (orbits, maxilla, zygoma, and nasal bones) sustained on the battlefield is needed to improve our understanding of these injuries, to optimize treatment, and to potentially direct strategic development of protective equipment in the future.

Experimental Verification of MicroRNA Targets is Essential, Prediction Alone is Insufficient

please see uploaded Letter-to-the-Editor file.

Proteomic Analysis of Temporally Stimulated Ovarian Cancer Cells for Biomarker Discovery

While ovarian cancer remains the most lethal gynecological malignancy in the United States, there are no biomarkers available that are able to predict therapeutic responses to ovarian malignancies. One major hurdle in the identification of useful biomarkers has been the ability to obtain enough ovarian cancer cells from primary tissues diagnosed in the early stages of serous carcinomas, the most deadly subtype of ovarian tumor. To detect ovarian cancer in a state of hyperproliferation, we analyzed the implications of molecular signaling cascades in the ovarian cancer cell line, OVCAR3, in a temporal manner, using a mass spectrometry-based proteomics approach. OVCAR3 cells were treated with EGF and the time course of cell progression was monitored by Akt phosphorylation and growth dynamics. EGF-stimulated Akt phosphorylation was detected at 12 h post-treatment, but an effect on proliferation was not observed until 48 h post-exposure. Growth- stimulated cellular lysates were analyzed for protein profiles between treatment groups and across time points using iTRAQ-labeling and mass spectrometry. Protein response to EGF treatment was identified via iTRAQ analysis in EGF-stimulated lysates as compared to vehicle-treated specimens across the treatment time course. Validation studies were performed on one of the differentially regulated proteins, lysosomal-associated membrane protein 1 (LAMP-1), in human tissue lysates and ovarian tumor tissue sections. Further, tissue microarray analysis was performed to demarcate LAMP-1 expression across different stages of epithelial ovarian cancers. These data support the use of this approach for efficient identification of tissue based markers in tumor development related to specific signaling pathways. LAMP-1 is a promising biomarker in understanding the progression of EGF-stimulated ovarian cancers and may be useful in the prediction of treatment responses involving tyrosine kinase inhibitors or EGF receptor monoclonal antibodies.

A Curvilinear Nomogram of Peak Expiratory Flow Rate for the Young

Objective. Peak expiratory flow rates (PEFRs) differ among populations and between times. The new EU scale of the mini-Wright flow-meter has been introduced since 2004. This study updated the PEFR nomograms with the new scale for Chinese children and adolescents (aged 6-19 years) in Hong Kong. Methods. A convenience sample was recruited from 34 primary care practices (patients' companions/children) and four schools. Standardization workshops were run for the physicians, and the proper use of the flow-meter was demonstrated to students prior to the data collection. Brand new meters were used. For each sex, the linear regression model was used to determine the relationship between PEFR and the variables of age and body height. The open-source software PyNomo was used to generate the nomograms. Results. After excluding 66 participants with past/current history of respiratory tract diseases, heart disease, incomplete data, and poor effort, PEFRs were collected from 798 males and 794 females. The PEFR had a linear relationship with age but a curvilinear relationship with height. The regression equations for predicted PEFR were ln(PEFR) = 1.810256*ln(height) + 0.038297*age - 3.734139 for males and ln(PEFR) = 1.525509*ln(height) + 0.033275*age - 2.368592 for females. The corresponding nomograms were constructed. They were tested with 230 patients in primary care; 9.6% (12 males and 10 females) had PEFR less than the predicted value by ≥20%. Conclusion. The body height was a stronger determinant than age for PEFR. The predicted PEFR with these determinants bear a curvilinear relationship.

Liver Stiffness Measurement: Simplicity Is Prerequisite For Reliability

Brain Structural Abnormalities in Obsessive-compulsive Disorder: Converging Evidence from White Matter and Grey Matter

Specific cortico-striato-thalamic circuits are hypothesised to underlie the aetiology of obsessive-compulsive disorder (OCD). However, findings from neuroimaging studies have been inconsistent. In the current study, we attempted to provide a complete overview of structural alterations in OCD by conducting signed differential mapping (SDM) meta-analysis on grey matter and white matter studies of patients with OCD based on voxel-based morphometry (VBM) studies and diffusion tensor imaging (DTI) studies.

Bringing out the Leader in You-Malaysian Psychiatric Association Leadership Certification for Early Career Psychiatrists

Identifying Infants at High Risk of Peanut Allergy: The Learning Early About Peanut Allergy (LEAP) Screening Study

BACKGROUND: Peanut allergy (PA) is rare in countries in which peanuts are introduced early into infants' diets. Learning Early About Peanut Allergy (LEAP) is an interventional study aiming to assess whether PA can be prevented by oral tolerance induction. OBJECTIVE: We sought to characterize a population screened for the risk of PA. METHODS: Subjects screened for the LEAP interventional trial comprise the LEAP screening study cohort. Infants were aged 4 to 10 months and passed a prescreening questionnaire. RESULTS: This analysis includes 834 infants (mean age, 7.8 months). They were split into the following: group I, patients with mild eczema and no egg allergy (n = 118); group II, patients with severe eczema, egg allergy, or both but 0-mm peanut skin prick test (SPT) wheal responses (n = 542); group III, patients with severe eczema, egg allergy, or both and 1- to 4-mm peanut wheal responses (n = 98); and group IV, patients with greater than 4-mm peanut wheal responses (n = 76). Unexpectedly, many (17%) in group II had peanut-specific IgE sensitization (≥0.35 kU/L); 56% of group III were similarly sensitized. In contrast, none of the patients in group I and 91% of those in group IV had peanut-specific IgE sensitization. Sensitization on skin testing to peanut (SPT response of 1-4 mm vs 0 mm) was associated with egg allergy and severe eczema (odds ratio [OR], 2.31 [95% CI, 1.39-3.86] and 2.47 [95% CI, 1.14-5.34], respectively). Similar associations were observed with specific IgE sensitization. Black race was associated with a significantly higher risk of peanut-specific IgE sensitization (OR, 5.30 [95% CI, 2.85-9.86]). Paradoxically, for a given specific IgE level, black race was protective against cutaneous sensitization (OR, 0.15 [95% CI, 0.04-0.61]). CONCLUSION: Egg allergy, severe eczema, or both appear to be useful criteria for identifying high-risk infants with an intermediate level of peanut sensitization for entry into a PA prevention study. The relationship between specific IgE level and SPT sensitization needs to be considered within the context of race.

Reaction-based Small-molecule Fluorescent Probes for Chemoselective Bioimaging

The dynamic chemical diversity of elements, ions and molecules that form the basis of life offers both a challenge and an opportunity for study. Small-molecule fluorescent probes can make use of selective, bioorthogonal chemistries to report on specific analytes in cells and in more complex biological specimens. These probes offer powerful reagents to interrogate the physiology and pathology of reactive chemical species in their native environments with minimal perturbation to living systems. This Review presents a survey of tools and tactics for using such probes to detect biologically important chemical analytes. We highlight design criteria for effective chemical tools for use in biological applications as well as gaps for future exploration.

Recent Advances in Laser Tweezers Raman Spectroscopy (LTRS) for Label-free Analysis of Single Cells

Laser tweezers Raman spectroscopy (LTRS), a technique that integrates optical tweezers with confocal Raman spectroscopy, is a variation of micro-Raman spectroscopy that enables the manipulation and biochemical analysis of single biological particles in suspension. This article provides an overview of the LTRS method, with an emphasis on highlighting recent advances over the past several years in the development of the technology and several new biological and biomedical applications that have been demonstrated. A perspective on the future developments of this powerful cytometric technology will also be presented. (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).

Cortical Effects of Repetitive Finger Flexion- Vs. Extension-resisted Tracking Movements: A TMS Study

While the cortical effects of repetitive motor activity are generally believed to be task-specific, the task parameters which modulate these effects are incompletely understood. Since there are differences in the neural control of flexor vs. extensor muscles, the type of muscles involved in the motor task of interest may be one important parameter. In addition, the role each muscle plays in the task, such as whether or not it is the prime mover, is another potentially important task parameter. In the present study, use-dependent cortical plasticity was examined in healthy volunteers performing a robotic waveform tracking task with either the extensor digitorum communis (EDC) or flexor digitorum superficialis (FDS) acting as the prime mover. Transcranial magnetic stimulation (TMS) was used to measure corticospinal excitability (CE) and short-interval intracortical inhibition of lower- and higher-threshold corticospinal neurons (SICIL and SICIH, respectively) before and after a flexion- or extension-resisted finger tracking task. After repetitive performance of the tracking task there was a significant decrease in SICIL targeting the EDC, while no change in CE targeting EDC was observed. In contrast, the reverse pattern was observed in the FDS: a significant increase in CE with no change in SICIL. There was also a tendency toward increased SICIH targeting whichever muscle was acting as the prime mover, though this effect did not reach statistical significance. We conclude that there is a difference in patterns of use-dependent plasticity between extrinsic finger flexor and extensor muscles performing the same task.

Prospective Study of Family History and Colorectal Cancer Risk by Tumor LINE-1 Methylation Level

BackgroundBeyond known familial colorectal cancer (CRC) syndromes, the mechanisms underlying the elevated CRC risk associated with CRC family history remain largely unknown. A recent retrospective study suggests familial clustering of CRC with hypomethylation in long interspersed nucleotide element 1 (LINE-1). We tested the hypothesis that CRC family history might confer a higher risk of LINE-1 methylation-low CRC.MethodsUsing the Nurses' Health Study and the Health Professionals Follow-up Study, we prospectively examined the association between CRC family history and the risk of rectal and colon cancer (N = 1224) according to tumor LINE-1 methylation level by duplication method Cox proportional hazards regression. We examined microsatellite instability (MSI) status to exclude the influence of Lynch syndrome. All statistical tests were two-sided.ResultsThe association between CRC family history and non-MSI CRC risk differed statistically significantly by LINE-1 methylation level (P (heterogeneity) = .02). CRC family history was associated with a statistically significantly higher risk of LINE-1 methylation-low non-MSI cancer (multivariable hazard ratio [HR] = 1.68, 95% confidence interval [CI] = 1.19 to 2.38 for 1 vs 0 first-degree relatives with CRC; multivariable HR = 3.48, 95% CI = 1.59 to 7.6 for ≥2 vs 0 first-degree relatives with CRC; P (trend) < .001). In contrast, CRC family history was not statistically significantly associated with LINE-1 methylation-high non-MSI cancer (P (trend) = .35).ConclusionsThis molecular pathological epidemiology study shows that CRC family history is associated with a higher risk of LINE-1 methylation-low CRC, suggesting previously unrecognized heritable predisposition to epigenetic alterations. Additional studies are needed to evaluate tumor LINE-1 methylation as a molecular biomarker for familial cancer risk assessment.

A Retrospective Study Investigating the Rate of HER2 Discordance Between Primary Breast Carcinoma and Locoregional or Metastatic Disease

ABSTRACT: BACKGROUND: Overall survival of HER2 positive metastatic breast cancer patients has been significantly improved with inclusion of trastuzumab to chemotherapy. Several studies have demonstrated discordant HER2 status in the primary and metastatic tumour. However, rates of discordance vary considerably in published reports. METHODS: Information collected prospectively was analysed for all patients seen from 1999 to 2009 with primary breast cancer and who had biopsy of a local or distant recurrence. Patients were included if adequate tissue was available from both paired samples. Recurrent samples included fine needle aspirations, core and excisional biopsies. HER2 status in all paired samples was assessed by in-situ hybridisation by a single pathologist in a national reference laboratory. This was compared with HER2 immunohistochemistry results provided in the course of routine diagnosis at regional laboratories. RESULTS: In total, 157 patients with recurrent (n = 137; 87.3%) or synchronous primary and metastatic (n = 20; 12.7%) breast cancer had biopsy of the metastatic site. The study population comprised of 116 patients with adequate tissue in both primary and metastasis. The concordance between HER2 status of the paired samples by local immunohistochemistry testing and central in-situ hybridization were 78% and 99%, respectively. Only one patient demonstrated HER2 discordance -- primary lesion was positive whilst a metastatic site was negative. CONCLUSIONS: This single institution study demonstrated a low rate of HER2 discordance between primary and recurrent breast cancer as assessed by in-situ hybridisation. This contrasts to results reported by others, which may be explained by differences in study methodology, definition of recurrent disease samples and generally small numbers of patients assessed. Despite the current findings, the decision to obtain metastatic tissue for evaluation is influenced by other factors. These include disease-free interval, which may raise the possibility of a new malignancy and the accuracy of initial HER2 assessment of the primary tumour.

Technique for Chordae Replacement in Mitral Valve Repair

We describe a simple technique for implantation of neochordae in degenerative mitral regurgitation with elongated or ruptured chords. We have successfully performed 102 mitral valve repairs using this technique and achieved mitral valve competency with no more than trace regurgitation. The technique is easy to perform and reproducible.

Combined Guillain-Barré Syndrome and Myasthenia Gravis

Background: Guillain-Barré syndrome and myasthenia gravis both lead to muscle weakness but the two combined is uncommon. Detection of these entities can help identify forms of autoimmune neuromuscular diseases that may respond to immunotherapy. This report sought to characterize the clinical features of these two entities when combined. Methods: This report is of a case of combined Guillain-Barré syndrome and myasthenia gravis. The clinical features were analyzed and correlated to those published in English literature from 1960 to 2012. Ten reports and 12 cases, including the present case, were reviewed. Results: There were 12 patients (4 women and 8 men), aged 17 to 84years, with combined Guillain-Barré syndrome and myasthenia gravis. Four had post-infectious Guillain-Barré syndrome followed by the development of myasthenia gravis concurrently or concomitantly within one month. All cases had symptoms of ptosis and areflexia. The other common presentations were limb weakness, oculobulbar weakness, and respiratory involvement. Functional outcome was mentioned in 10 patients and seven had good outcome (Hughes scale ⩽2). Conclusion: Detection of ptosis with or without ophthalmoplegia, distribution of limb weakness, and reflex can help in recognizing combined Guillain-Barré syndrome and myasthenia gravis. The early recognition of this combination of peripheral nervous and neuro-muscular junction inflammation is important for initial treatment and prognosis.

Joint Practice Guideline for Sterile Technique During Vascular and Interventional Radiology Procedures: From the Society of Interventional Radiology, Association of PeriOperative Registered Nurses, and Association for Radiologic and Imaging Nursing, for the Society of Interventional Radiology (Wael Saad, MD, Chair), Standards of Practice Committee, and Endorsed by the Cardiovascular Interventional Radiological Society of Europe and the Canadian Interventional Radiology Association

Using Bioactive Glasses for the Treatment of Periodontal Intrabony Defects May Result in Modest Probing Depth Reduction and Clinical Attachment Level Gain Compared to Open Flap Debridement

ARTICLE TITLE AND BIBLIOGRAPHIC INFORMATION: An evaluation of bioactive glass in the treatment of periodontal defects: a meta-analysis of randomized controlled clinical trials. Sohrabi K, Saraiya V, Laage TA, Harris M, Blieden M, Karimbux N. J Periodontol 2012;83:453-64. REVIEWERS: Hsun-Liang Chan, DDS, MS, Hom-Lay Wang, DDS, PhD* PURPOSE/QUESTION: To determine whether the use of bioactive glass in the treatment of periodontal intrabony defects results in more PD reduction and CAL gain than other treatment methods SOURCE OF FUNDING: Information not available TYPE OF STUDY/DESIGN: Systematic review with meta-analysis LEVEL OF EVIDENCE: Level 2: limited-quality, patient-oriented evidence STRENGTH OF RECOMMENDATION GRADE: Grade C: Consensus, disease-oriented evidence, usual practice, expert opinion, or case series for studies of diagnosis, treatment, prevention, or screening.

Hypomethylation of the IL17RC Promoter Associates with Age-Related Macular Degeneration

Age-related macular degeneration (AMD) is the leading cause of irreversible blindness in the elderly population worldwide. Although recent studies have demonstrated strong genetic associations between AMD and SNPs in a number of genes, other modes of regulation are also likely to play a role in the etiology of this disease. We identified a significantly decreased level of methylation on the IL17RC promoter in AMD patients. Furthermore, we showed that hypomethylation of the IL17RC promoter in AMD patients led to an elevated expression of its protein and messenger RNA in peripheral blood as well as in the affected retina and choroid, suggesting that the DNA methylation pattern and expression of IL17RC may potentially serve as a biomarker for the diagnosis of AMD and likely plays a role in disease pathogenesis.

13-year Long-term Associations Between Changes in Traditional Cardiovascular Risk Factors and Changes in Fibrinogen Levels: The Coronary Artery Risk Development in Young Adults (CARDIA) Study

OBJECTIVE: Cross-sectional and prospective studies have linked cardiovascular events and traditional risk factors (TRFs) with higher plasma fibrinogen levels. In a young cohort, we sought to determine longitudinal associations between changes in/development of TRFs and fibrinogen levels over 13 years. METHODS: We included 2525 adults from the CARDIA study, aged 25-37 with fibrinogen and TRFs measured at year 7 (study baseline; 1992-1993); and year 20 (follow-up). Multiple linear regressions were used to compare mean changes in fibrinogen to TRFs. RESULTS: Mean fibrinogen increased by 71 mg/dL vs. 70 mg/dL (p = NS) in black vs. white men, and 78 mg/dL vs. 68 mg/dL (p < 0.05) in black vs. white women, respectively over 13 years. After multivariable adjustments, fibrinogen generally rose with increasing BMI (p < 0.001; all sex/race groups), LDL cholesterol, log triglycerides and diastolic blood pressure; and fell with increasing HDL cholesterol and physical activity. 13-year increase in fibrinogen for persons who quit smoking or became non-obese were comparable (p = NS) to that of never-smokers and never-obese persons. CONCLUSIONS: Among young black and white men and women with few baseline cardiovascular risk factors, fibrinogen tracked longitudinally with changes in TRFs over 13 years through middle age. There was a strong inverse longitudinal relationship between modifiable risk factors (weight loss/smoking cessation) and 13-year change in fibrinogen. Our study helps provide some insight into the role of fibrinogen as a disease marker in the associations between fibrinogen and CVD.

Ovarian Malignancy Risk Stratification of the Adnexal Mass Using a Multivariate Index Assay

OBJECTIVE: To validate the effectiveness of a multivariate index assay in identifying ovarian malignancy compared to clinical assessment and CA125-II, among women undergoing surgery for an adnexal mass after enrollment by non-gynecologic oncology providers. METHODS: A prospective, multi-institutional trial enrolled female patients scheduled to undergo surgery for an adnexal mass from 27 non-gynecologic oncology practices. Pre-operative serum samples and physician assessment of ovarian cancer risk were correlated with final surgical pathology. RESULTS: A total of 494 subjects were evaluable for multivariate index assay, CA125-II, and clinical impression. Overall, 92 patients (18.6%) had a pelvic malignancy. Primary ovarian cancer was diagnosed in 65 patients (13.2%), with 43.1% having FIGO stage I disease. For all ovarian malignancies, the sensitivity of the multivariate index assay was 95.7% (95%CI=89.3-98.3) when combined with clinical impression. The multivariate index assay correctly predicted ovarian malignancy in 91.4% (95%CI=77.6-97.0) of cases of early-stage disease, compared to 65.7% (95%CI=49.2-79.2) for CA125-II. The multivariate index assay correctly identified 83.3% malignancies missed by clinical impression and 70.8% cases missed by CA125-II. Multivariate index assay was superior in predicting the absence of an ovarian malignancy, with a negative predictive value of 98.1% (95%CI=95.2-99.2). Both clinical impression and CA125-II were more accurate at identifying benign disease. The multivariate index assay correctly predicted benign pathology in 204 patients (50.7%, 95%CI=45.9-55.6) when combined with clinical impression. CONCLUSION: The multivariate index assay demonstrated higher sensitivity and negative predictive value for ovarian malignancy compared to clinical impression and CA125-II in an intended-use population of non-gynecologic oncology practices.

Comparison of N-linked Glycosylation of Protein C in Newborns and Adults

Protein C (PC) is a major anticoagulant that stems the propagation of thrombin. The activated form of PC (APC), in association with the cofactor protein S, proteolytically converts activated coagulation factors VIIIa and Va into inactive forms. Studies have shown that forms of PC that contain 3N-linked glycans (beta-PC) are functionally distinct from the fully glycosylated 4-glycan type (alpha-PC). Since some findings have also hinted at qualitative differences in PC from newborns and adults, we decided to determine the relative constitution of glycoforms in these age groups. Subtypes of PC in newborn and adult plasmas were distinguished by SDS polyacrylamide electrophoresis and Western blotting, followed by immunological analysis. Newborns were found to have alpha-PC/beta-PC mole ratios of 8.8:1, compared to 2.3:1 in adults. PC was also isolated by immunoaffinity chromatography from newborn and adult plasmas. Glycans were released by protease treatment and studied by mass spectrometry. Results from glycan analysis showed a small range of glycan structures in both age groups. No clear differences were noted between newborn and adult PC microheterogeneity in glycan structures (branching). We conclude that newborns have important differences in PC macroheterogeneity in glycoform content relative to adults. This age-dependent glycosylation variation may have implications in management of PC function in vivo.

Hyaluronic Acid-dependent Protection in H9C2 Cardiomyocytes: A Cell Model of Heart Ischemia-reperfusion Injury and Treatment

Hyaluronic acid (HA), a glycosaminoglycan with high molecular weight, has been reported to promote cell proliferation and serves as an important extracellular matrix component. The aim of this study was to in vitro investigate whether HA is able to reduce reactive oxygen species (ROS)-induced heart ischemia-reperfusion injury and activate the cardiomyocyte's damage surveillance systems. Accordingly, rattus cardiomyocyte line, H9C2, was treated with H(2)O(2) as a heart ischemia-reperfusion model followed by incubation with low molecular weight hyaluronan (LMW-HA, 100kDa) or high molecular weight hyaluronan (HMW-HA, 1000kDa) and proteomic analysis was performed to investigate the physiologic protection of HA in H(2)O(2)-induced ischemia-reperfusion in cardiomyocyte. Our data demonstrated that HA treatment does protect cardiomyocyte in the ROS-induced ischemia-reperfusion model and the molecular weight of HA is a crucial factor. HMW-HA has been shown to significantly facilitate cell migration and wound healing via cytoskeletal rearrangement. Additionally, 2D-DIGE combined MALDI-TOF/TOF analysis showed that HMW-HA might modulate biosynthetic pathways, cell migration, cell outgrowth and protein folding to stimulate wound healing as well as prevent these ischemia-reperfusion-damaged cardiomyocytes from cell death. To our knowledge, we report for the first time the cell repair mechanism of HMW-HA against ischemia-reperfusion-damage in cardiomyocytes based on cell biology and proteomic analysis.

Increased Anti-leukemic Activity of Decitabine Via AR-42-induced Upregulation of MiR-29b: A Novel Epigenetic-targeting Approach in Acute Myeloid Leukemia

Histone deacetylase (HDAC) inhibitors either alone or in combination with hypomethylating agents have limited clinical effect in acute myeloid leukemia (AML). Previously we demonstrated that AML patients with higher miR-29b expression had better response to the hypomethylating agent decitabine. Therefore, an increase in miR-29b expression preceding decitabine treatment may provide a therapeutic advantage. We previously showed that miR-29b expression is suppressed by a repressor complex that includes HDACs. Thus, HDAC inhibition may increase miR-29b expression. We hypothesized that priming AML cells with the novel HDAC inhibitor (HDACI) AR-42 would result in increased response to decitabine treatment via upregulation of miR-29b. Here we show that AR-42 is a potent HDACI in AML, increasing miR-29b levels and leading to downregulation of known miR-29b targets (i.e., SP1, DNMT1, DNMT3A, and DNMT3B). We then demonstrated that the sequential administration of AR-42 followed by decitabine resulted in a stronger anti-leukemic activity in vitro and in vivo than decitabine followed by AR-42 or either drug alone. These preclinical results with AR-42 priming before decitabine administration represents a promising, novel treatment approach and a paradigm shift with regard to the combination of epigenetic-targeting compounds in AML, where decitabine has been traditionally given before HDAC inhibitors.Leukemia accepted article preview online, 26 November 2012; doi:10.1038/leu.2012.342.

The Predictive Validity of OMPQ on the Rehabilitation Outcomes for Patients with Acute and Subacute Non-Specific LBP in a Chinese Population

Introduction Early screening of physical and psychosocial risk factors has been advocated as a way to identify low back pain (LBP) patients who may develop chronic disability. This study evaluated the predictive validity of a Chinese version of the Orebro Musculoskeletal Pain Questionnaire (OMPQ) in identifying LBP patients at risk of developing poor return-to-work (RTW) outcomes. Methods Altogether 241 patients with acute or subacute non-specific LBP agreed to participate, and they were screened at baseline with OMPQ, and evaluated after discharge from physiotherapy (n = 173) with outcome measures including the Roland-Morris Disability Questionnaire (RMDQ), numerical pain score (0-10) and global recovery (0-10). At 1-year follow-up, information on RTW status as well as sick leave duration were obtained. Results At baseline the OMPQ had a mean score of 112.0 (SD = 26.5). The receiver operator characteristic (ROC) curves of OMPQ scores at 1-year follow-up recorded values of area under the curve of 0.693 for RTW and 0.714 for sick leave duration, which are comparable to those reported in European studies. OMPQ was the only factor that could significantly predict the RTW outcomes, compared to other variables such as the RMDQ scores. Conclusion The results confirmed the predictive validity of the Chinese version of OMPQ in screening LBP patients at risk of developing poor occupational outcomes, and appropriate interventions can be arranged for these high-risk individuals in the rehabilitation process.

Single Laboratory Evaluation of a Planar Waveguide-based System for a Simple Simultaneous Analysis of Four Mycotoxins in Wheat

The accuracy and precision of a commercially available system based on an indirect competitive immunoassay and planar waveguide technology was evaluated for the analysis of deoxynivalenol (DON), ochratoxin A (OTA), zearalenone (ZEAR), and T-2 toxin in wheat. The system generally performed well at the tested concentrations that were close to the regulatory limits of DON and OTA in wheat. The mean percent recovery of OTA from certified and in-house reference materials ranged from 90 to 111 %, with a relative standard deviation of 8-16 % (at 4.2, 4.9, and 7.0 μg/kg). Mean percent recoveries of DON ranged from 75 to 103 %, with a relative standard deviation of 14-20 % (at 610, 940, and 1300 μg/kg). As analyte concentrations approached the lower limits of the working range of 3 μg/kg OTA and 400 μg/kg DON, the mean percent recoveries and relative standard deviation increased for both DON and OTA. A lack of reference materials precluded a thorough evaluation of the method for the analysis of ZEAR and T-2. The particular strength of the technology was that multiple mycotoxins were analyzed simultaneously.

Prospective Study on Serum Metal Levels in Patients with Metal-on-metal Lumbar Disc Arthroplasty

PURPOSE: Metal-on-metal total disc replacement is a recent alternative treatment for degenerative disc disease. Wear and corrosion of these implants can lead to local and systemic transport of metal debris. This prospective longitudinal study examined the serum chromium and cobalt levels in 24 patients with cobalt-chromium alloy metal-on-metal lumbar disc replacements. METHODS: Serum was assayed for chromium (Cr) and cobalt (Co) using high-resolution inductively-coupled plasma-mass spectrometry. Detection limits were 0.015 ng/mL for Cr and 0.04 ng/mL for Co. RESULTS: Median serum Co levels at pre-op, 3, 6, 12, 24, and 36-months post-op were 0.10, 1.03, 0.96, 0.98, 0.67, and 0.52 ng/mL, respectively. Median serum Cr levels were 0.06, 0.49, 0.65, 0.43, 0.52, and 0.50 ng/mL, respectively. CONCLUSION: In general, these results indicated that serum Co and Cr levels are elevated at all postoperative time points and are of the same order of magnitude as those observed in well-functioning metal-on-metal surface replacements of the hip and in metal-on-metal total hip replacements at similar postoperative time points.

Optimizing Hollow-fiber-based Pharmacokinetic Assay Via Chemical Stability Study to Account for Inaccurate Simulated Drug Clearance of Rifampicin

With increasing multidrug resistance coupled to a poor development pipeline, clinicians are exploring antimicrobial combinations to improve treatment outcomes. In vitro hollow-fiber infection model (HFIM) is employed to simulate human in vivo drug clearance and investigate pharmacodynamic synergism of antibiotics. Our overarching aim was to optimize the HFIM-based pharmacokinetic (PK) assay by using rifampicin and polymyxin B as probe drugs. An ultrapressure liquid chromatography tandem mass spectrometry method was validated for the quantification of rifampicin and polymyxin B components. In vitro profiling studies demonstrated that the experimental PK profiles of polymyxin B monotherapy were well correlated with the human population PK data while monotherapy with rifampicin failed to achieve the expected maximum plasma concentration. Chemical stability studies confirmed polymyxin B was stable in broth at 37 °C up to 12 h while rifampicin was unstable under the same conditions over 12 and 80 h. The calculated mean clearance of rifampicin due to chemical degradation was 0.098 ml/min accounting for 12.2 % of its clinical total clearance (CL = 0.8 ml/min) based on population PK data. Our novel finding reinforces the importance to optimize HFIM-based PK assay by performing chemical stability study so as to account for potential discrepancy between experimental and population PK profiles of antimicrobial agents.

In Vivo Quantification of Active Decitabine-Triphosphate Metabolite: A Novel Pharmacoanalytical Endpoint for Optimization of Hypomethylating Therapy in Acute Myeloid Leukemia

Decitabine (DAC) is used for treatment of patients with myelodysplastic syndromes and acute myeloid leukemia (AML). Following cellular uptake, DAC is activated to DAC-triphosphate (TP) and incorporated into DNA. Once incorporated into the DNA, DAC-TP binds and inactivates DNA methyltransferases (DNMTs), thereby leading to hypomethylation and re-expression of epigenetically silenced tumor suppressor genes and ultimately antileukemia activity. However, direct evidence of in vivo DAC-TP occurrence in DAC-treated patients has been difficult to demonstrate due to a lack of suitable validated analytical methodology. Thus, we developed and validated a nonradioactive sensitive and specific LC-MS/MS assay for quantification of DAC-TP. The assay is linear from 50 to 1,000 nM and from 1 to 10 μM and has a lower limit of quantitation of 50 nM and a coefficient of variation for both within- and between-day precision <20%. Following DAC treatment, we detected DAC-TP in parental and DAC-resistant AML cells (in vitro) and bone marrow (BM) and spleen of normal and leukemic mice (in vivo). Downregulation of DNMTs and correlation of DAC-TP concentration with proteins involved in mechanisms of DAC resistance were also demonstrated. The clinical applicability of this method was proven by measuring DAC-TP level in BM and blood mononuclear cells from DAC-treated AML patients. Higher levels are seemingly associated with clinical response. Monitoring the DAC-TP intracellular level may serve as a novel pharmacological endpoint for designing more effective DAC-based regimens.

Overshadowing of Geometric Cues by a Beacon in a Spatial Navigation Task

In three experiments, we examined whether overshadowing of geometric cues by a discrete landmark (beacon) is due to the relative saliences of the cues. Using a virtual water maze task, human participants were required to locate a platform marked by a beacon in a distinctively shaped pool. In Experiment 1, the beacon overshadowed geometric cues in a trapezium, but not in an isosceles triangle. The longer escape latencies during acquisition in the trapezium control group with no beacon suggest that the geometric cues in the trapezium were less salient than those in the triangle. In Experiment 2, we evaluated whether generalization decrement, caused by the removal of the beacon at test, could account for overshadowing. An additional beacon was placed in an alternative corner. For the control groups, the beacons were identical; for the overshadow groups, they were visually unique. Overshadowing was again found in the trapezium. In Experiment 3, we tested whether the absence of overshadowing in the triangle was due to the geometric cues being more salient than the beacon. Following training, the beacon was relocated to a different corner. Participants approached the beacon rather than the trained platform corner, suggesting that the beacon was more salient. These results suggest that associative processes do not fully explain cue competition in the spatial domain.

Fetal Cardiac Catheterization Using a Percutaneous Transhepatic Access Technique: Preliminary Experience in a Lamb Model

Objectives: Human fetal cardiac intervention has typically involved a percutaneous transventricular approach. In fetal lambs, a transhepatic approach to access the fetal intra-abdominal veins after exteriorization of the uterus by laparotomy has been described. We aimed to develop a percutaneous transhepatic technique for catheterization of the fetal heart at mid-gestation that avoids maternal laparotomy. Methods: In 10 fetal lambs (90-97 days gestation), access to the fetal venous system was by percutaneous puncture with a 5Fr sheath (n = 1) into the umbilical vein or 16G IV-catheter (n = 9) into the hepatic vein. This was followed by cardiac catheterization using a 1.8-2.6Fr tapered coronary catheter. Euthanasia and postmortem examination were planned immediately post-procedure (n = 2), or after normal term-delivery (remaining cases). Results: In 1 case fetal position precluded procedural attempts, and in another, the fetus, accessed by a 5Fr sheath, died from umbilical haemorrhage. In 8 cases, access to the fetal hepatic vein was achieved. In 7 of these cases the access catheter was advanced into the IVC, followed by catheterization of the RA (all cases) and 4 cardiac chambers (3 cases). One fetus died during cardiac catheterization (RV perforation) and the other 7 fetuses were alive at the end of the procedure (88% survival). Immediate postmortem (n = 2) detected intraperitoneal bleeding (4 and 20ml respectively), while the postnatal postmortem (n = 5) after uneventful pregnancy revealed no vascular or cardiac trauma. Conclusions: Ultrasound-guided percutaneous transhepatic cardiac catheterization is feasible in mid-gestational fetal sheep. This technique has the potential for translation into human fetal cardiac and circulatory interventions. Copyright © 2012 ISUOG. Published by John Wiley & Sons, Ltd.

The Origins and Genetic Distinctiveness of the Chamorros of the Marianas Islands: An MtDNA Perspective

BACKGROUND: Archaeological and linguistic evidence suggests the Marianas Islands were settled around 3,600 years before present (ybp) from Island Southeast Asia (ISEA). Around 1,000 ybp latte stone pillars and the first evidence of rice cultivation appear in the Marianas. Both traditions are absent in the rest of prehistoric Oceania. OBJECTIVE: To examine the genetic origins and postsettlement gene flow of Chamorros of the Marianas Islands. METHODS: To infer the origins of the Chamorros we analyzed ∼360 base pairs of the hypervariable-region 1 (HVS1) of mitochondrial DNA from 105 Chamorros from Guam, Rota, and Saipan, and the complete mitochondrial genome of 32 Guamanian Chamorros, and compared them to lineages from ISEA and neighboring Pacific archipelagoes from the database. RESULTS: Results reveal that 92% of Chamorros belong to haplogroup E, also found in ISEA but rare in Oceania. The two most numerous E lineages were identical to lineages currently found in Indonesia, while the remaining E lineages differed by only one or two mutations and all were unique to the Marianas. Seven percent of the lineages belonged to a single Chamorro-specific lineage within haplogroup B4, common to ISEA as well as Micronesia and Polynesia. CONCLUSIONS: These patterns suggest a small founding population had reached and settled the Marianas from ISEA by 4,000 ybp, and developed unique mutations in isolation. A second migration from ISEA may have arrived around 1,000 ybp, introducing the latte pillars, rice agriculture and the homogeneous minority B4 lineage. Am. J. Hum. Biol., 2012. © 2012Wiley Periodicals, Inc.

Communication: Excited States, Dynamic Correlation Functions and Spectral Properties from Full Configuration Interaction Quantum Monte Carlo

In this communication, we propose a method for obtaining isolated excited states within the full configuration interaction quantum Monte Carlo framework. This method allows for stable sampling with respect to collapse to lower energy states and requires no uncontrolled approximations. In contrast with most previous methods to extract excited state information from quantum Monte Carlo methods, this results from a modification to the underlying propagator, and does not require explicit orthogonalization, analytic continuation, transient estimators, or restriction of the Hilbert space via a trial wavefunction. Furthermore, we show that the propagator can directly yield frequency-domain correlation functions and spectral functions such as the density of states which are difficult to obtain within a traditional quantum Monte Carlo framework. We demonstrate this approach with pilot applications to the neon atom and beryllium dimer.

Visual Observations of an Atmospheric-pressure Solution-cathode Glow Discharge

The solution-cathode glow discharge (SCGD) is an optical emission source for atomic spectrometry comprised of a moderate-power atmospheric-pressure DC glow discharge sustained directly upon the surface of an electrically conductive solution. The SCGD boasts a simple, inexpensive design and has demonstrated detection limits similar to those of more conventional excitation sources used in atomic spectrometry. Although the analytical performance of the SCGD as an optical emission source is well characterized, the mechanism through which the discharge atomizes and excites analyte from the sample solution remains a point of debate. The current paper presents visual observations of the SCGD from a variety of imaging techniques. The implications of the images regarding the mechanism of analyte solution-to-plasma transport and excitation in the SCGD are discussed.

Management of Hereditary Angioedema with C1-inhibitor Concentrate During Two Successive Pregnancies

Gene Expression-Based Model Using Formalin-Fixed Paraffin-Embedded Biopsies Predicts Overall Survival in Advanced-Stage Classical Hodgkin Lymphoma

PURPOSEOur aim was to reliably identify patients with advanced-stage classical Hodgkin lymphoma (cHL) at increased risk of death by developing a robust predictor of overall survival (OS) using gene expression measured in routinely available formalin-fixed paraffin-embedded tissue (FFPET). METHODSExpression levels of 259 genes, including those previously reported to be associated with outcome in cHL, were determined by digital expression profiling of pretreatment FFPET biopsies from 290 patients enrolled onto the E2496 Intergroup trial comparing doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) and Stanford V regimens in locally extensive and advanced-stage cHL. A model for OS separating patients into low- and high-risk groups was produced using penalized Cox regression. The model was tested in an independent cohort of 78 patients enriched for treatment failure but otherwise similar to patients in a population-based registry of patients treated with ABVD. Weighted analysis methods generated unbiased estimates of predictor performance in the population-based registry.ResultsA 23-gene outcome predictor was generated. The model identified a population at increased risk of death in the validation cohort. There was a 29% absolute difference in 5-year OS between the high- and low-risk groups (63% v 92%, respectively; log-rank P < .001; hazard ratio, 6.7; 95% CI, 2.6 to 17.4). The predictor was superior to the International Prognostic Score and CD68 immunohistochemistry in multivariate analyses. CONCLUSIONA gene expression-based predictor, developed in and applicable to routinely available FFPET biopsies, identifies patients with advanced-stage cHL at increased risk of death when treated with standard-intensity up-front regimens.

Current Controversies in Breast Cancer Surgery

Contemporary management of the axilla in breast cancer surgery remains in evolution. Axillary lymph node status in breast cancer is a major prognostic factor and remains integral to guiding adjuvant treatment decisions. There remains controversy regarding the management of the node-positive axilla in clinically node-negative primary breast cancer. Trials to date have suggested re-evaluation of the historical therapeutic strategy that a positive sentinel node requires axillary node dissection. However, further evidence is required before modern clinical management of the axilla should be altered. As patient awareness and technical expertise grow, national rates of breast reconstruction after mastectomy continue to rise. Oncoplastic techniques continue to evolve and many patients are suitable for a plethora of reconstructive options. Despite the widespread practice of breast reconstruction globally, there is limited randomised evidence comparing the optimal type and/or timing of breast reconstruction on which to base practice. Breast reconstruction type is either purely autologous, implant-based or a combination of these two techniques. We explore the benefits and limitations of these techniques and some of the key findings of the National Mastectomy and Breast Reconstruction Audit. The timing of reconstruction after mastectomy is either immediate (a single procedure) or delayed (for an indefinite period after mastectomy). The ideal reconstruction is one that is best aligned to the patient's expectations, as this will achieve the highest levels of long-term patient satisfaction. Selecting the optimal type of breast reconstruction at the right time for the right patient remains the key challenge in breast reconstruction.

Tumor Inside a Pearl Drop

The confined internal space of a liquid marble, as well as its porous and non-adhesive shell, offers an attractive application possibility - accommodating living cells inside liquid marbles. Cancer cells in suspension may aggregate to form three dimensional structures, also known as cancer cell spheroids (CCS). In this study, CCS formation inside liquid marble is investigated. This liquid marble application opens significant and novel avenues for biomedical applications and cancer research.

Fluorescence-tagged Gold Nanoparticles for Rapidly Characterizing the Size-dependent Biodistribution in Tumor Models

Nanoparticle vehicles may improve the delivery of contrast agents and therapeutics to diseased tissues, but their rational design is currently impeded by a lack of robust technologies to characterize their in vivo behavior in real-time. This study demonstrates that fluorescent-labeled gold nanoparticles can be optimized for in vivo detection, perform pharmacokinetic analysis of nanoparticle designs, analyze tumor extravasation, and clearance kinetics in tumor-bearing animals. This optical imaging approach is non-invasive and high-throughput. Interestingly, these fluorescent gold nanoparticles can be used for multispectral imaging to compare several nanoparticle designs simultaneously within the same animal and eliminates the host-dependent variabilities across measured data. Together these results describe a novel platform for evaluating the performance of tumor-targeting nanoparticles, and provide new insights for the design of future nanotherapeutics.

Compression-induced Alignment and Elongation of Human Mesenchymal Stem Cell (hMSC) in 3D Collagen Constructs is Collagen Concentration Dependent

Controlling cell organization is important in tissue engineering. Guidance by aligned features on scaffolds or stimulation by physical signals can be used to induce cell alignment. We have previously demonstrated a preferred alignment of human MSCs (hMSCs) along the compression loading axis in 3D collagen construct. In this study, we aim to investigate the collagen concentration dependence of the compression-induced hMSC organization. Results demonstrated that the compression-induced alignment and elongation of hMSCs exhibited a biphasic dose-dependent relationship with collagen concentration, and associated well with both collagen ligand density and elastic modulus of the constructs. Moreover, collagen concentration and compression loading significantly affected the expression level of integrin beta 1 and antibody neutralization against this molecule aborted the compression-induced alignment and elongation responses. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2012.

Correction: Discovery of a Natural Product-Like C-myc G-Quadruplex DNA Groove-Binder by Molecular Docking

[This corrects the article on p. e43278 in vol. 7.].

Six-Month Mortality Among HIV-Infected Adults Presenting for Antiretroviral Therapy with Unexplained Weight Loss, Chronic Fever or Chronic Diarrhea in Malawi

In sub-Saharan Africa, early mortality is high following initiation of antiretroviral therapy (ART). We investigated 6-month outcomes and factors associated with mortality in HIV-infected adults being assessed for ART initiation and presenting with weight loss, chronic fever or diarrhea, and with negative TB sputum microscopy.

Ezrin Interacts with the SARS Coronavirus Spike Protein and Restrains Infection at the Entry Stage

Entry of Severe Acute Respiratory Syndrome coronavirus (SARS-CoV) and its envelope fusion with host cell membrane are controlled by a series of complex molecular mechanisms, largely dependent on the viral envelope glycoprotein Spike (S). There are still many unknowns on the implication of cellular factors that regulate the entry process.

A Metal-Based Inhibitor of NEDD8-Activating Enzyme

A cyclometallated rhodium(III) complex [Rh(ppy)(2)(dppz)](+) (1) (where ppy = 2-phenylpyridine and dppz = dipyrido[3,2-a:2',3'-c]phenazine dipyridophenazine) has been prepared and identified as an inhibitor of NEDD8-activating enzyme (NAE). The complex inhibited NAE activity in cell-free and cell-based assays, and suppressed the CRL-regulated substrate degradation and NF-κB activation in human cancer cells with potency comparable to known NAE inhibitor MLN4924. Molecular modeling analysis suggested that the overall binding mode of 1 within the binding pocket of the APPBP1/UBA3 heterodimer resembled that for MLN4924. Complex 1 is the first metal complex reported to suppress the NEDDylation pathway via inhibition of the NEDD8-activating enzyme.

Rhodopsin Gene Expression Determines Rod Outer Segment Size and Rod Cell Resistance to a Dominant-negative Neurodegeneration Mutant

Two outstanding unknowns in the biology of photoreceptors are the molecular determinants of cell size, which is remarkably uniform among mammalian species, and the mechanisms of rod cell death associated with inherited neurodegenerative blinding diseases such as retinitis pigmentosa. We have addressed both questions by performing an in vivo titration with rhodopsin gene copies in genetically engineered mice that express only normal rhodopsin or an autosomal dominant allele, encoding rhodopsin with a disease-causing P23H substitution. The results reveal that the volume of the rod outer segment is proportional to rhodopsin gene expression; that P23H-rhodopsin, the most common rhodopsin gene disease allele, causes cell death via a dominant-negative mechanism; and that long term survival of rod cells carrying P23H-rhodopsin can be achieved by increasing the levels of wild type rhodopsin. These results point to promising directions in gene therapy for autosomal dominant neurodegenerative diseases caused by dominant-negative mutations.

Genetic Association Studies in Lumbar Disc Degeneration: a Systematic Review

Low back pain is associated with lumbar disc degeneration, which is mainly due to genetic predisposition. The objective of this study was to perform a systematic review to evaluate genetic association studies in lumbar disc degeneration as defined on magnetic resonance imaging (MRI) in humans.

Lipocalin 2 Regulates Inflammation During Pulmonary Mycobacterial Infections

Pulmonary tuberculosis (TB), caused by the intracellular bacteria Mycobacterium tuberculosis, is a worldwide disease that continues to kill more than 1.5 million people every year worldwide. The accumulation of lymphocytes mediates the formation of the tubercle granuloma in the lung and is crucial for host protection against M.tuberculosis infection. However, paradoxically the tubercle granuloma is also the basis for the immunopathology associated with the disease and very little is known about the regulatory mechanisms that constrain the inflammation associated with the granulomas. Lipocalin 2 (Lcn2) is a member of the lipocalin family of proteins and binds to bacterial siderophores thereby sequestering iron required for bacterial growth. Thus far, it is not known whether Lcn2 plays a role in the inflammatory response to mycobacterial pulmonary infections. In the present study, using models of acute and chronic mycobacterial pulmonary infections, we reveal a novel role for Lcn2 in constraining T cell lymphocytic accumulation and inflammation by inhibiting inflammatory chemokines, such as CXCL9. In contrast, Lcn2 promotes neutrophil recruitment during mycobacterial pulmonary infection, by inducing G-CSF and KC in alveolar macrophages. Importantly, despite a common role for Lcn2 in regulating chemokines during mycobacterial pulmonary infections, Lcn2 deficient mice are more susceptible to acute M.bovis BCG, but not low dose M.tuberculosis pulmonary infection.

Development and Validation of Chinese Health Literacy Scale for Diabetes

AIMS AND OBJECTIVES: To develop and to test the psychometric properties of the Chinese Health Literacy Scale for Diabetes. BACKGROUND: Patients with diabetes encounter many challenges when making health decisions in their daily lives, as they have access to many different kinds of health information. Health literacy issues are new topics in Chinese society. Without a valid and reliable instrument in Chinese, it is difficult to measure the level of health literacy and promote the concept of health literacy in Chinese societies. DESIGN: A methodological study with a sample of 137 patients with type 2 diabetes aged 65 years or older. METHOD: Chinese Health Literacy Scale for Diabetes was developed with reference to the revised Bloom's taxonomy model. Psychometric tests (content validity, item analysis, construct validity, discriminative ability and test-retest reliability) were conducted. Correlations between Chinese Health Literacy Scale for Diabetes and four relevant measures were tested. Cronbach's alpha and alpha if item deleted were calculated to assess internal consistency. RESULTS: Cronbach's alpha for Chinese Health Literacy Scale for Diabetes and its four subscales (remembering, understanding, applying and analysing) were 0·884, 0·885, 0·667, 0·654 and 0·717, respectively. The Chinese Health Literacy Scale for Diabetes was significantly correlated with the Diabetic Knowledge Scale (r = 0·398, p < 0·001), the Diabetic Management Self-Efficacy Scale (r = 0·257, p < 0·001), the Preschool and Primary Chinese Literacy Scale (r = 0·822, p < 0·001) and the Chinese Value of Learning Scale (r = 0·303, p < 0·001). It took an average of seven minutes to complete this 34-item instrument. CONCLUSION: The findings of this study support the Chinese Health Literacy Scale for Diabetes as a reliable and valid instrument for measuring the health literacy of Chinese patients with diabetes. RELEVANCE TO CLINICAL PRACTICE: We recommend that clinicians use this tool to assess patients' health literacy before conducting any kind of health promotion.

The Pharmacoeconomics of Managing Acute Agitation in the Emergency Department: What Do We Know and How Do We Approach It?

The emergency department (ED) is commonly the first point of care for patients with acute behavioral issues from the community. Routinely, clinical management involves the use of benzodiazepine and/or antipsychotic drugs, when initial de-escalation strategies fail. There is currently scant literature available to inform the clinical management and resource utilization of acute agitation in the ED. This article discusses the approach to pharmacoeconomic studies of acute agitation management in the ED. It explores the conduct of such evaluations and highlights the cost and data sources required. The current difficulties experienced in conducting such evaluations are also discussed. Pharmacoeconomic studies related to the management of acute agitation in ED can be challenging. Robust clinical trials incorporating prospectively designed pharmacoeconomic studies will invariably contribute toward a better understanding of this therapeutic area and optimize the use of scarce resources.

Electroluminescence from ZnO-nanofilm/Si-micropillar Heterostructure Arrays

ZnO-nanofilm/Si-micropillar p-n nanoheterostructure arrays were prepared by growing n-type ZnO onto a p-type nanoporous Si pillar array. Its current-voltage characteristics of nanoheterostructure showed good rectifying behavior with onset voltage of ~1.5 V, forward current density of ~28.7 mA/cm2 at 2.5 V, leakage current density of ~0.15 mA/cm2 and rectifying ratio of ~121 at ± 2.5 V. The electron transport across nanohetreostructure obeys the trap-charge-limit current model. Moreover, strong white light electroluminescence from ZnO-nanofilm/Si-micropillar light-emitting diode (LED) has been achieved, which could open up possibilities to build new ZnO/Si-based highly efficient solid-state lighting devices.

Time-variant 1D Photonic Crystals Using Flowing Microdroplets

In this paper we propose a time-variant photonic crystal, which can be formed by a stream of wave-length-scale microdroplets flowing through a microfluidic channel. The functionality stems from the photonic bandgap generated from the 1D periodic perturbation of refractive index. The periodicity, volume fraction and composition of both the dispersed and the continuous phases can be conveniently tuned in real time by hydrodynamic or pneumatic methods. By simulation, it is found that the time-variant nature of the proposed structure can induce an abnormal energy evolution, which is distinct from any existing photonic crystal structures. As a basic component for optofluidic systems, the droplet-based photonic crystal may find potential applications in light modulation and light confinement, and could be an ideal model for pursuing physical insights into time-variant optofluidic systems.

Slot-mode-coupled Optomechanical Crystals

We present a design methodology and analysis of a cavity optomechanical system in which a localized GHz frequency mechanical mode of a nanobeam resonator is evanescently coupled to a high quality factor (Q > 106) optical mode of a separate nanobeam optical cavity. Using separate nanobeams provides flexibility, enabling the independent design and optimization of the optics and mechanics of the system. In addition, the small gap (≈25 nm) between the two resonators gives rise to a slot mode effect that enables a large zero-point optomechanical coupling strength to be achieved, with g/2π > 300 kHz in a Si3N4 system at 980 nm and g/2π ≈ 900 kHz in a Si system at 1550 nm. The fact that large coupling strengths to GHz mechanical oscillators can be achieved in Si3N4 is important, as this material has a broad optical transparency window, which allows operation throughout the visible and near-infrared. As an application of this platform, we consider wide-band optical frequency conversion between 1300 nm and 980 nm, using two optical nanobeam cavities coupled on either side to the breathing mode of a mechanical nanobeam resonator.

Regulatory Focus Influences Perseveration and Distractibility in Task Switching

In two experiments, we investigated the effect of regulatory focus (Higgins, Am Psychol, 52, 1280-1300, 1997) on people's maintenance and switching behavior. In the experimental tasks, participants were asked to react selectively to one category of stimuli while ignoring the other. After several practice trials, participants had to switch either to new stimuli which were paired with former target stimuli (i.e., in the perseveration condition), or switch to former distracter stimuli which were paired with new stimuli (i.e., in the learned irrelevance condition). Results from both experiments indicated that a promotion focus promoted switching to new stimuli in the perseveration condition, at the cost of poorer performance on switching to former distracters in the learned irrelevance condition. This pattern of results applied to both chronic individual differences in regulatory focus (Experiment 1), and regulatory focus temporarily manipulated in the laboratory (Experiment 2). It suggests that whereas a promotion focus indeed promotes cognitive flexibility, it also incurs a cost in terms of increased likelihood of being distracted.

Volumetric Intensity-modulated Arc Therapy Vs Conventional Intensity-modulated Radiation Therapy in Nasopharyngeal Carcinoma: a Dosimetric Study

Dosimetric comparisons between RapidArc (RA) and conventional Intensity-Modulated Radiation Therapy (IMRT) techniques for nasopharyngeal carcinoma (NPC) were performed to address differences in dose coverage of the target, sparing of organs-at-risk (OARs), delivery of monitor units (MUs) and time, to assess whether the RA technique was more beneficial for treatment of NPC. Eight NPC patients (Stages I-IV), who had completed RA treatment, were selected for this study. Computed tomography data sets were re-planned using 7-fields fixed beam IMRT. Quantitative measurements of dose-endpoint values on the dose-volume histograms were carried out for evaluation of: (i) dose homogeneity (D(5%) - D(95%)); (ii) degree of conformity (CI(95%)); (iii) tumor control probability (TCP); (iv) doses to OARs; (v) normal tissue complication probability (NTCP); (vi) treatment time; and (vii) MUs. RA plans achieved better dose conformity and TCP in planning target volumes (PTVs). Target dose homogeneity was not as high as for IMRT plans. Doses to tempero-mandibular joints, clavicles, parotid glands and posterior neck, and their NTCPs were significantly lower in RA plans (P < 0.05). Mean doses to the brainstem and spinal cord were slightly lower in IMRT plans. RA plans allowed for a mean reduction in MUs by 78% (P = 0.006), and a four-fold reduction in treatment delivery times, relative to IMRT plans. RA plans showed superior, or comparable, target coverage and dose conformity in PTVs, but at the expense of inferior dose homogeneity. RA plans also achieved significant improvements in dose reduction to OARs and healthy tissue sparing. A significant reduction in treatment delivery time for RA treatment technique was also noted.

The Laboratory Risk Indicator for Necrotizing Fasciitis Score for Discernment of Necrotizing Fasciitis Originated from Vibrio Vulnificus Infections

BACKGROUND: The Laboratory Risk Indicator for Necrotizing Fasciitis (LRINEC) score has been verified as a useful diagnostic tool for detecting necrotizing fasciitis (NF). Its application, however, is mainly for NF types I and II. The practical relevance of the LRINEC score for Vibro vulnificus-related skin and soft tissue infection (SSTI) was hardly ever investigated. The aim of this study was to assess the applicability of the LRINEC scoring system and to identify NF-predicting factors in patients with V. vulnificus-caused SSTI. METHODS: A retrospective study was conducted, enrolling 125 consecutive patients diagnosed with V. vulnificus-related SSTI who were admitted to a teaching hospital between January 2003 and December 2011. Demographics, laboratory data, comorbidities, treatment, and outcomes were collected for each patient and extracted for analysis. Logistic regression and receiver operating characteristic curve analyses were performed. RESULTS: The mean (SD) age of the 125 patients was 63.0 (10.9) years; 58% of the patients were male. The mean (SD) LRINEC score at admission was 2.4 (1.9) points. Of the 125 patents, 72 (58%) had NF. Multivariate analysis revealed that the presence of hemorrhagic bullous lesions (p = 0.002) and higher LRINEC scores at admission (p < 0.0001) were significantly associated with the presence of NF. In addition, the area under the receiver operating characteristic curve for the LRINEC scoring model for detecting NF was 0.783 (p < 0.0001). An optimal cutoff LRINEC score of 2 or greater had a sensitivity of 71%, a specificity of 83%, and a positive predictive value of 85%, with an 11.9-fold increased risk for the presence of NF (p < 0.0001). CONCLUSION: We have demonstrated that the LRINEC score and hemorrhagic bullous/blistering lesions are significant predictors of NF in patients with V. vulnificus-related SSTI. V. vulnificus-infected patients having hemorrhagic bullous/blistering lesions or with an LRINEC score of 2 or greater should be thoughtfully evaluated for the presence of NF. LEVEL OF EVIDENCE: Diagnostic test study, level II.

Tunable Coherence-free Microwave Photonic Bandpass Filter Based on Double Cross Gain Modulation Technique

A tunable, coherence-free, high-resolution microwave photonic bandpass filter, which is compatible to be inserted in a conventional fiber optic link, is presented. It is based on using two cross gain modulation based wavelength converters in a recursive loop. The double cross gain modulation technique solves the semiconductor optical amplifier facet reflection problem in the conventional recursive structure; hence the new microwave photonic signal processor has no coherent interference and no phase-induced intensity noise. It allows arbitrary narrow-linewidth telecommunication-type lasers to be used while enabling stable filter operation to be realized. The filter passband frequency can be tuned by using a wavelength tunable laser and a wavelength dependent time delay component. Experimental results demonstrate robust high-resolution bandpass filter operation with narrow-linewidth sources, no phase-induced intensity noise and a high signal-to-noise ratio performance. Tunable coherence-free operation of the high-resolution bandpass filter is also demonstrated.

A New Technique for 100-fold Increase in the FSR of Optical Recirculating Delay Line Filters Using a Time Compression Unit

A new technique that increases the free spectral range (FSR) of a recirculating delay line filter, is presented. The concept is based on a time-compression unit, which is used in conjunction with a frequency-shifting recirculating loop that generates multi-spectral characteristics, and the idea exploits the optical wavelength domain by wavelength-to-time mapping of the taps using an oppositely time-oriented dispersive element so that the taps travel different lengths, to time compress the tap separation. This technique solves, for the first time, the long-standing problem of the small FSR limitation in recirculating microwave photonic delay line filters, opening the way to realize the main functionalities required in microwave photonic filters. Experimental results are presented which demonstrate a large 100-fold increase in the FSR of the bandpass filter response.

Mental Health Risks in the Local Workforce Engaged in Disaster Relief and Reconstruction

To build a sustainable workforce for long-term disaster relief and reconstruction, more effort must be made to promote local relief workers' mental health. We conducted 25 semistructured interviews with local relief officials 10 months after the 2008 earthquake in China to investigate the stress and coping experiences in their personal lives as survivors. We conducted thematic analysis of interview transcripts. Traumatic bereavement and grief, housing and financial difficulties, and work-family conflict were the three main sources of stress in the respondents' personal lives. The coping themes were finding meaning and purpose in life through relief work, colleagues' support and understanding, suppression or avoidance of grief, appreciation for life, hardiness, optimism, letting nature take its course, and making up for loss. We suggest that relief work has a double-edged-sword effect on workers' coping abilities. We discuss the implications of this effect for work-life balance measures and trauma and grief counseling services.

Pharmacological Reduction of NEFA Restores the Efficacy of Incretin-based Therapies Through GLP-1 Receptor Signalling in the Beta Cell in Mouse Models of Diabetes

AIMS/HYPOTHESIS: Type 2 diabetes mellitus is associated with reduced incretin effects. Although previous studies have shown that hyperglycaemia contributes to impaired incretin responses in beta cells, it is largely unknown how hyperlipidaemia, another feature of type 2 diabetes, contributes to impaired glucagon-like peptide 1 (GLP-1) response. Here, we investigated the effects of NEFA on incretin receptor signalling and examined the glucose-lowering efficacy of incretin-based drugs in combination with the lipid-lowering agent bezafibrate. METHODS: We used db/db mice to examine the in vivo efficacy of the treatment. Beta cell lines and mouse islets were used to examine GLP-1 and glucose-dependent insulinotropic peptide receptor signalling. RESULTS: Palmitate treatment decreased Glp1r expression in rodent insulinoma cell lines and isolated islets. This was associated with impairment of the following: GLP-1-stimulated cAMP production, phosphorylation of cAMP-responsive elements binding protein (CREB) and insulin secretion. In insulinoma cell lines, the expression of exogenous Glp1r restored cAMP production and the phosphorylation of CREB. Treatment with bezafibrate in combination with des-fluoro-sitagliptin or exendin-4 led to more robust glycaemic control, associated with improved islet morphology and beta cell mass in db/db mice. CONCLUSIONS/INTERPRETATION: Elevated NEFA contributes to impaired responsiveness to GLP-1, partially through downregulation of GLP-1 receptor signalling. Improvements in lipid control in mouse models of obesity and diabetes increase the efficacy of incretin-based therapy.

Buddhism and Medical Futility

Religious faith and medicine combine harmoniously in Buddhist views, each in its own way helping Buddhists enjoy a more fruitful existence. Health care providers need to understand the spiritual needs of patients in order to provide better care, especially for the terminally ill. Using a recently reported case to guide the reader, this paper examines the issue of medical futility from a Buddhist perspective. Important concepts discussed include compassion, suffering, and the significance of the mind. Compassion from a health professional is essential, and if medical treatment can decrease suffering without altering the clarity of the mind, then a treatment should not be considered futile. Suffering from illness and death, moreover, is considered by Buddhists a normal part of life and is ever-changing. Sickness, old age, birth, and death are integral parts of human life. Suffering is experienced due to the lack of a harmonious state of body, speech, and mind. Buddhists do not believe that the mind is located in the brain, and, for Buddhists, there are ways suffering can be overcome through the control of one's mind.

A Phase II, Randomized, Controlled Trial of Acupuncture for Reduction of Postcolectomy Ileus

PURPOSE: Postoperative ileus (POI) prolongs hospital stay and increases risk of postoperative complications. We conducted a randomized, sham-controlled trial to evaluate whether acupuncture reduces POI more effectively than sham acupuncture. METHODS: Colon cancer patients undergoing elective colectomy were randomized to receive 30 min of true or sham acupuncture twice daily during their first 3 postoperative days. GI-3 (the later of the following two events: time that the patient first tolerated solid food, AND time that the patient first passed flatus OR a bowel movement) and GI-2 (the later of the following two events: time patient first tolerated solid food AND time patient first passed a bowel movement) were determined. Pain, nausea, vomiting, and use of pain medications were evaluated daily for the first 3 postoperative days. RESULTS: Ninety patients were randomized. Eighty-one received the allocated intervention: 39 in the true acupuncture group and 42 in the sham acupuncture group, all evaluated for the primary endpoint. The mean time to GI-3 was 149 h [standard deviation (SD) 71 h] and 146 (SD 62 h) after surgery for the acupuncture group and the sham acupuncture group (difference between means -2 h; 95 % confidence interval -31, 26; p = 0.9). No significant differences were found between groups for secondary endpoints. CONCLUSIONS: True acupuncture as provided in this study did not reduce POI more significantly than sham acupuncture. The study was limited by a standard deviation much larger than expected, suggesting that a study with a larger sample size might be required.

Burn Injury Triggered Dysfunction in Dendritic Cell Response to TLR9 Activation and Resulted in Skewed T Cell Functions

Severe trauma such as burn injury is often associated with a systemic inflammatory syndrome characterized by a hyperactive innate immune response and suppressed adaptive immune function. Dendritic cells (DCs), which sense pathogens via their Toll-like receptors (TLRs), play a pivotal role in protecting the host against infections. The effect of burn injury on TLR-mediated DC function is a debated topic and the mechanism controlling the purported immunosuppressive response remains to be elucidated. Here we examined the effects of burn injury on splenic conventional DC (cDC) and plasmacytoid DC (pDC) responses to TLR9 activation. We demonstrate that, following burn trauma, splenic cDCs' cytokine production profile in response to TLR9 activation became anti-inflammatory dominant, with high production of IL-10 (>50% increase) and low production of IL-6, TNF-α and IL-12p70 (∼25-60% reduction). CD4+ T cells activated by these cDCs were defective in producing Th1 and Th17 cytokines. Furthermore, burn injury had a more accentuated effect on pDCs than on cDCs. Following TLR9 activation, pDCs displayed an immature phenotype with an impaired ability to secrete pro-inflammatory cytokines (IFN-α, IL-6 and TNF-α) and to activate T cell proliferation. Moreover, cDCs and pDCs from burn-injured mice had low transcript levels of TLR9 and several key molecules of the TLR signaling pathway. Although hyperactive innate immune response has been associated with severe injury, our data show to the contrary that DCs, as a key player in the innate immune system, had impaired TLR9 reactivity, an anti-inflammatory phenotype, and a dysfunctional T cell-priming ability. We conclude that burn injury induced impairments in DC immunobiology resulting in suppression of adaptive immune response. Targeted DC immunotherapies to promote their ability in triggering T cell immunity may represent a strategy to improve immune defenses against infection following burn injury.

Marginal Bone Loss Around Tilted Implants in Comparison to Straight Implants: a Meta-analysis

Purpose: The primary aim of this systematic review was to compare the amount of marginal bone loss around tilted and straight implants. As the secondary aim, the incidence of biomechanic complications was compared. Materials and Methods: An electronic literature search from five databases, for the years 2000 to 2011, and a hand search in implant-related journals were conducted. Clinical human studies in the English language that had reported marginal bone loss in tilted and straight implants at 12-months follow-up or longer were included. Mean marginal bone loss and the number of implants that were available for analysis were extracted from original articles for meta-analyses. Results: Eight (six prospective and two retrospective) studies were included. One-year data were available in seven articles, which included 1,015 (451 tilted) implants. Three articles provided 3- to 5-year data from 302 (164 tilted) implants. No significant difference in weighted mean marginal bone loss was found between the tilted and straight implants in the short and medium terms. Three articles reported the incidence of biomechanic complications. There was not enough information to make a comparison. Conclusions: This meta-analysis failed to support the hypothesis that tilted implants that were splinted for the support of fixed prostheses had more marginal bone loss. Additionally, there was not enough evidence to claim a higher incidence of biomechanic complications in tilted implants. However, due to the nature of the study design of the included articles, caution should be exercised when interpreting the results of this review.

Interview with Chi-chao Chan

Chi-Chao Chan, an American board certified ophthalmologist, is the Chief of Immunopathology Section, Laboratory of Immunology and the Head of Histology Core at the National Eye Institute at National Institutes of Health (USA). In 1967, Dr Chan graduated from Chungzhan Medical College in Guangzhou, China and then earned her AB (1972), as well as MD (1975) at Johns Hopkins University (USA). Currently, her research is focusing on primary intraocular (vitreoretinal) lymphoma; uveitis (intraocular inflammatory diseases); and the molecular pathology, animal models, and genetic epidemiology of age-related macular degeneration. Dr Chan spoke to Future Medicinal Chemistry, about the challenges ophthalmic research faces and how the aging population could lead to better drugs. Interview conducted by Isaac Bruce, Commissioning Editor.

Pathogenic Cellular Phenotypes Are Germline Transmissible in a Transgenic Primate Model of Huntington's Disease

A transgenic primate model for Huntington's Disease (HD) first reported by our group that (HD monkeys) carry the mutant Huntingtin (HTT) gene with expanded polyglutamine (CAG) repeats and, develop chorea, dystonia and other involuntary motor deficiencies similar to Huntington's disease (HD) [1]. More recently, we have found that longitudinal magnetic resonance imaging (MRI) of the HD monkey brain revealed significant atrophy in regions associated with cognitive deficits symptomatic in HD patients, providing the first animal model which replicates clinical phenotypes of diagnosed humans. Here we report germline transmission of the pathogenic mutant HTT in HD monkey by the production of embryos and subsequent derivation of HD monkey embryonic stem cells (rHD-ESCs) using HD monkey sperm. rHD-ESCs inherit mutant HTT and green fluorescent protein (GFP) genes through the gametes of HD monkey. rHD-ESCs express mutant HTT and form intranuclear inclusion, a classical cellular feature of HD. Notably, mosaicism of the pathogenic polyQ region in the sperm as well as derived ESCs was also observed, consistent with intraindividual and intergenerational reports of mosaic CAG repeats [2, 3]and CAG expansion in HD patients [4-7]. The confirmation of transgene inheritability and development of pathogenic HD phenotype in derived rHD-ESCs reported in this study is a milestone in the pursuit of a transgenic primate model with inherited mutant HTT for development of novel disease biomarkers and therapeutics.

Dose-distance Metric That Predicts Late Rectal Bleeding in Patients Receiving Radical Prostate External-beam Radiotherapy

The relationship between rectal dose distribution and the incidence of late rectal complications following external-beam radiotherapy has been previously studied using dose-volume histograms or dose-surface histograms. However, they do not account for the spatial dose distribution. This study proposes a metric based on both surface dose and distance that can predict the incidence of rectal bleeding in prostate cancer patients treated with radical radiotherapy. One hundred and forty-four patients treated with radical radiotherapy for prostate cancer were prospectively followed to record the incidence of grade ≥2 rectal bleeding. Radiotherapy plans were used to evaluate a dose-distance metric that accounts for the dose and its spatial distribution on the rectal surface, characterized by a logistic weighting function with slope a and inflection point d(0). This was compared to the effective dose obtained from dose-surface histograms, characterized by the parameter n which describes sensitivity to hot spots. The log-rank test was used to determine statistically significant (p < 0.05) cut-off values for the dose-distance metric and effective dose that predict for the occurrence of rectal bleeding. For the dose-distance metric, only d(0) = 25 and 30 mm combined with a > 5 led to statistical significant cut-offs. For the effective dose metric, only values of n in the range 0.07-0.35 led to statistically significant cut-offs. The proposed dose-distance metric is a predictor of rectal bleeding in prostate cancer patients treated with radiotherapy. Both the dose-distance metric and the effective dose metric indicate that the incidence of grade ≥2 rectal bleeding is sensitive to localized damage to the rectal surface.

A Luminescent G-quadruplex Switch-on Probe for the Highly Selective and Tunable Detection of Cysteine and Glutathione

A G-quadruplex-selective luminescent iridium(iii) switch-on probe has been developed for the detection of cysteine (Cys) in aqueous solution. The system is highly sensitive and selective towards Cys with a tunable range of detection. The detection of glutathione (GSH) is also examined.

Quantifying Limb Movements in Epileptic Seizures Through Color-based Video Analysis

This paper proposes a color-based video analytic system for quantifying limb movements in epileptic seizure monitoring. The system utilizes colored pyjamas to facilitate limb segmentation and tracking. Thus, it is unobtrusive and requires no sensor/marker attached to patients body. We employ Gaussian mixture models in background/foreground modeling, and detect limbs through a coarse-to-fine paradigm with graphcut- based segmentation. Next, we estimate limb parameters with domain knowledge guidance, and extract displacement and oscillation features from movement trajectories for seizure detection/analysis. We report studies on sequences captured in an epilepsy monitoring unit. Experimental evaluations show that the proposed system has achieved comparable performance to EEG-based systems in detecting motor seizures.

Predicting Basal Metabolic Rates in Malaysian Adult Elite Athletes

Introduction: This study aimed to measure the basal metabolic rate (BMR) of elite athletes and develop a genderspecific predictive equation to estimate their energy requirements. Methods: 92 men and 33 women (aged 18-31 years) from 15 sports, who had been training six hours daily for at least one year, were included in the study. Body composition was measured using the bioimpedance technique, and BMR by indirect calorimetry. The differences between measured and estimated BMR using various predictive equations were calculated. The novel equation derived from stepwise multiple regression was evaluated using Bland and Altman analysis. Results: The predictive equations of Cunningham and the Food and Agriculture Organization/World Health Organization/United Nations University either over- or underestimated the measured BMR by up to ± 6%, while the equations of Ismail et al, developed from the local non-athletic population, underestimated the measured BMR by 14%. The novel predictive equation for the BMR of athletes was BMR (kcal/day) = 669 + 13 (weight in kg) + 192 (gender: 1 for men and 0 for women) (R2 0.548; standard error of estimates 163 kcal). Predicted BMRs of elite athletes by this equation were within 1.2% ± 9.5% of the measured BMR values. Conclusion: The novel predictive equation presented in this study can be used to calculate BMR for adult Malaysian elite athletes. Further studies may be required to validate its predictive capabilities for other sports, nationalities and age groups.

Successful Nutritional Therapy for Superior Mesenteric Artery Syndrome

Superior mesenteric artery (SMA) syndrome is an uncommon cause of duodenal outlet obstruction. Symptoms and signs suggestive of this condition are nonspecific, and a high index of suspicion coupled with appropriate imaging studies are necessary for diagnosis. We present the case of a 70-year-old man who developed SMA syndrome following prolonged hospitalisation for a surgically treated bleeding duodenal ulcer. His SMA syndrome resolved after successful nonoperative management based on accepted guidelines for nutritional therapy, thus avoiding the need for reoperation and its attendant risks in a malnourished patient.

Fetal Aneuploidy Screening by Maternal Plasma DNA Sequencing: 'False Positive' Due to Confined Placental Mosaicism

High Dynamic Range Organic Temperature Sensor

A thermistor-transistor integrated organic temperature sensor is reported. By inserting a thin layer of silver nanoparticles the organic thermistor shows strong temperature sensitivity and the total dynamic range of sensing is around 10 bits which is larger than most organic temperature sensors. The measuring temperature range is from 20 to 70 °C with operating voltage down to 6 V.

Resonance Raman Based Skin Carotenoid Measurements in Newborns and Infants

We describe Resonance Raman based skin carotenoid measurements in newborns and infants. Skin- and serum carotenoid levels correlate with high statistical significance in healthy newborns and infants, and with reduced accuracy also in prematurely born infants, who in general feature very low carotenoid levels and thin transparent skin giving rise to large background absorption effects. Skin carotenoid levels can be easily compared among subjects and/or tracked in longitudinal studies with the highly molecule-specific Raman method. It therefore holds promise as a rapid, non-invasive, carotenoid antioxidant assessment method for newborns and infants in the field of pediatrics. (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).

Standardization of Post-thrombotic Syndrome Definition and Outcome Assessment Following Upper Venous System Thrombosis in Pediatric Practice

A Case of Sexual Abuse by a Traditional Faith Healer: Are There Potential Preventions?

Adolescent sexual abuse is not an uncommon phenomenon in Malaysia. It is a traumatic experience that complicates the psychosocial development of young people on the threshold of adulthood. This case report highlights the psychosocial sequelae of adolescent sexual abuse by a traditional healer and discusses management issues in the context of unique cultural and belief systems.

Neurophysiological Pharmacodynamic Measures of Groups and Individuals Extended from Simple Cognitive Tasks to More "lifelike" Activities

OBJECTIVE: Extend a method to track neurophysiological pharmacodynamics during repetitive cognitive testing to a more complex "lifelike" task. METHODS: Alcohol was used as an exemplar psychoactive substance. An equation, derived in an exploratory analysis to detect alcohol's EEGs effects during repetitive cognitive testing, was validated in a Confirmatory Study on a new group whose EEGs after alcohol and placebo were recorded during working memory testing and while operating an automobile driving simulator. RESULTS: The equation recognized alcohol by combining five times beta plus theta power. It worked well (p<.0001) when applied to both tasks in the confirmatory group. The maximum EEG effect occurred 2-2.5h after drinking (>1h after peak BAC) and remained at 90% at 3.5-4h (BAC<50% of peak). Individuals varied in the magnitude and timing of the EEG effect. CONCLUSION: The equation tracked the EEG response to alcohol in the Confirmatory Study during both repetitive cognitive testing and a more complex "lifelike" task. The EEG metric was more sensitive to alcohol than several autonomic physiological measures, task performance measures or self-reports. SIGNIFICANCE: Using EEG as a biomarker to track neurophysiological pharmacodynamics during complex "lifelike" activities may prove useful for assessing how drugs affect integrated brain functioning.

Care Managers' Experiences in a Collaborative Care Program for High Risk Mothers with Depression

OBJECTIVES: The aim of this study was to understand care managers' experiences in caring for depressed mothers in an integrated behavioral health program. METHODS: As part of a quality improvement project, we conducted a focus group interview with six care managers caring for low income mothers with behavioral health needs in a safety net program in King County, WA. Using thematic analysis, codes were organized into themes that described the care managers' experiences. RESULTS: Two organizing themes along with associated themes emerged: (1) Assets for improving depression outcomes: patient-provider interactions, including the importance of engagement; program resources such as care coordination and (2) Barriers to improved depression outcomes: patient-provider interactions, including difficulty engaging patient; patient-related factors such as multiple stressors; program resources such as need for more psychiatric support; and difficulty accessing outside resources. CONCLUSIONS:: Numerous potentially modifiable factors including levels of engagement, motivational interviewing, and increased psychiatric support were identified by care managers as affecting depression care and outcomes. Implications for care management training and approaches to psychiatric consultations are discussed.

Reply: Ethical Responsibility to Disclose Surgical Errors

Controversial View of a Genetically Altered Mouse Model of Focal Retinal Degeneration

Tuo et al. (2012) demonstrated tumor necrosis factor-inducible gene 6 recombinant protein (TSG-6) arrest of focal retinal lesions on a Ccl2 and Cx3cr1 double deficient mouse (DKO) on rd8 background (hereon referred to as DKO rd8). DKO rd8, a model of focal retinal degeneration with earlier onset and higher penetrance than Ccl2 and Cx3cr1 single knockout strains, demonstrates characteristic features of AMD such as focal photoreceptor atrophy, retinal pigmented epithelium (RPE) degeneration, elevated ocular A2E levels and complement deposition in addition to retinal dystrophy. The discovery of the accidently introduced Crb1 mutation (rd8) in the C57BL/6N strain has led to the recent opinion that DKO rd8 is not a model of AMD but solely a model of Crb1‑associated retinal degeneration. Differences between DKO rd8 and Crb1 (rd8) photoreceptor and RPE pathology, as well as increased A2E and immune dysfunction, show that DKO rd8 recapitulates some AMD‑like features in addition to rd8 retinal dystrophy. The appearance of rd8 lesions and Ccl2/Cx3cr1 lesions and the amelioration of most Ccl2/Cx3cr1 lesions in intervention studies show DKO rd8 to be a useful and appropriate model for therapeutic compound screening, such as the case with anti-inflammatory TSG‑6.

A Simple and Reliable Immunohistochemical Method for Colocalization of 2 Antigens in the Same Cells of Paraffin-embedded Tissues

Immunohistochemistry (IHC) lacks an efficient technique for colocalizing multiple antigens in the same cells of a single tissue section. The development of a methodology which combines the advantage of low cost, high sensitivity, and specificity would benefit clinical diagnosis and general research. On the basis of a newly published method of visualizing 2 antigens on a single paraffin-embedded tissue section, we have further developed a novel sequential technique for colocalizing 2 different antigens in a same cell in a paraffin-embedded tissue section. In this technique, we combined the microwave heating technique (MVT) with normal IHC methods to sequentially double stain a paraffin section; and colocalize 2 antigens in a single cell through result comparison stored in a digital management system. This MVT colocalization method has a higher degree of sensitivity and specificity comparable with conventional staining of both immunofluorescence and IHC systems. The primary advantage of this method is that it is inexpensive and convenient; the antibody(s) used in this method can be generated from the same or different species; it allows colocalization or comparison of different results of cell morphology for any single cell of the section on 2 images, avoids uncertainty when overlapping 2 antigens on a single image.

Cost Effectiveness of Full Coverage of the Medical Management of Smoking Cessation in France

OBJECTIVE: To estimate the incremental cost effectiveness of full coverage of the medical management of smoking cessation from the perspective of statutory health insurance (SHI) in France. DESIGN AND POPULATION: Cost-effectiveness analysis based on a Markov state-transition decision analytic model was used to compare full SHI coverage of smoking cessation and actual coverage based on an annual €50 lump sum per insured person among current French smokers aged 15-75 years. We used a scenario approach to take into account the many different behaviours of smokers and the likely variability of SHI policy choices in terms of participation rate and number and frequency of attempts covered. INTERVENTIONS: Drug treatments for smoking cessation combined with six medical consultations including individual counselling. MAIN OUTCOMES MEASURES: The cost effectiveness of full coverage was expressed by the incremental cost-effectiveness ratio (ICER) in 2009 euros per life-year gained (LYG) at the lifetime horizon. RESULTS: The cost effectiveness per LYG for smokers ranged from €1786 to €2012, with an average value of €1911. The minimum value was very close to the maximum value with a difference of only €226. The cost-effectiveness ratio was only minimally sensitive to the participation rate, the number of attempts covered and the cessation rate. CONCLUSIONS: Compared to other health measures in primary and secondary prevention of cardiovascular disease already covered by SHI, full coverage of smoking cessation is the most cost-effective approach.

Sustained Expression of the Transcription Factor GLIS3 is Required for Normal Beta Cell Function in Adults

Genome-wide association studies identified GLIS3 as a susceptibility locus for type 1 and type 2 diabetes. Global Glis3 deficiency in mice leads to congenital diabetes and neonatal lethality. In this study, we explore the role of Glis3 in adulthood using Glis3(+/-) and conditional knockout animals. We challenged Glis3(+/-) mice with high fat diet for 20 weeks and found that they developed diabetes because of impaired beta cell mass expansion. GLIS3 controls beta cell proliferation in response to high-fat feeding at least partly by regulating Ccnd2 transcription. To determine if sustained Glis3 expression is essential to normal beta cell function, we generated Glis3(fl/fl) /Pdx1Cre(ERT+) animal by intercrossing Glis3(fl/fl) mice with Pdx1Cre(ERT+) mice and used tamoxifen (TAM) to induce Glis3 deletion in adults. Adult Glis3(fl/fl) /Pdx1Cre(ERT+) mice are euglycaemic. TAM-mediated beta cell-specific inactivation of Glis3 in adult mice downregulates insulin expression, leading to hyperglycaemia and subsequently enhanced beta cell apoptosis. We conclude that normal Glis3 expression is required for pancreatic beta cell function and mass maintenance during adulthood, which impairment leads to diabetes in adults.

SRPK2 Phosphorylates Tau and Mediates the Cognitive Defects in Alzheimer's Disease

Serine-arginine protein kinases 2 (SRPK2) is a cell cycle-regulated kinase that phosphorylates serine/arginine domain-containing proteins and mediates pre-mRNA splicing with unclear function in neurons. Here, we show that SRPK2 phosphorylates tau on S214, suppresses tau-dependent microtubule polymerization, and inhibits axonal elongation in neurons. Depletion of SRPK2 in dentate gyrus inhibits tau phosphorylation in APP/PS1 mouse and alleviates the impaired cognitive behaviors. The defective LTP in APP/PS1 mice is also improved after SRPK2 depletion. Moreover, active SRPK2 is increased in the cortex of APP/PS1 mice and the pathological structures of human Alzheimer's disease (AD) brain. Therefore, our study suggests SRPK2 may contribute to the formation of hyperphosphorylated tau and the pathogenesis of AD.

Setting Priorities for Development of Emerging Interventions Against Childhood Pneumonia, Meningitis and Influenza

Setting Research Priorities to Reduce Global Mortality from Preterm Birth and Low Birth Weight by 2015

This paper aims to identify health research priorities that could improve the rate of progress in reducing global neonatal mortality from preterm birth and low birth weight (PB/LBW), as set out in the UN's Millennium Development Goal 4.

Social, Economic, Political and Health System and Program Determinants of Child Mortality Reduction in China Between 1990 and 2006: A Systematic Analysis

Between 1990 and 2006, China reduced its under-five mortality rate (U5MR) from 64.6 to 20.6 per 1000 live births and achieved the fourth United Nation's Millennium Development Goal nine years ahead of target. This study explores the contribution of social, economic and political determinants, health system and policy determinants, and health programmes and interventions to this success.

Sacrococcygeal Teratoma in an Adolescent

Sacrococcygeal teratoma (SCT) is a neoplasm derived from more than one primitive germ cell layer. SCTs have rarely been reported in adults. We present a case of a 17-year-old youth who complained of a painful mass over his left buttock.The mass had been growing gradually since his childhood. Magnetic resonance imaging showed a well-defined soft-tissue mass which contained mainly fat tissue and large intestine. The patient had complete excision of the tumor and coccyx.The gross impression and histology revealed a mature teratoma.

Emissive Behavior, Cytotoxic Activity, Cellular Uptake, and PEGylation Properties of New Luminescent Rhenium(I) Polypyridine Poly(ethylene Glycol) Complexes

We report here a new class of biological reagents derived from luminescent rhenium(I) polypyridine complexes modified with a poly(ethylene glycol) (PEG) pendant. The PEG-amine complexes [Re(N(∧)N)(CO)(3)(py-PEG-NH(2))](PF(6)) (py-PEG-NH(2) = 3-amino-5-(N-(2-(ω-methoxypoly(1-oxapropyl))ethyl)aminocarbonyl)pyridine, MW(PEG) = 5000 Da, PDI(PEG) < 1.08; N(∧)N = 1,10-phenanthroline (phen) (1-PEG-NH(2)), 3,4,7,8-tetramethyl-1,10-phenanthroline (Me(4)-phen) (2-PEG-NH(2)), 4,7-diphenyl-1,10-phenanthroline (Ph(2)-phen) (3-PEG-NH(2))) and [Re(bpy-PEG)(CO)(3)(py-NH(2))](PF(6)) (bpy-PEG = 4-(N-(2-(ω-methoxypoly(1-oxapropyl))ethyl)aminocarbonyl)-4'-methyl-2,2'-bipyridine; py-NH(2) = 3-aminopyridine) (4-PEG-NH(2)) have been synthesized and characterized. The photophysical properties, lipophilicity, water solubility, cytotoxic activity, and cellular uptake properties of these complexes have been compared to those of their PEG-free counterparts [Re(N(∧)N)(CO)(3)(py-Et-NH(2))](PF(6)) (py-Et-NH(2) = 3-amino-5-(N-(ethyl)aminocarbonyl)pyridine; N(∧)N = phen (1-Et-NH(2)), Me(4)-phen (2-Et-NH(2)), Ph(2)-phen (3-Et-NH(2))) and [Re(bpy-Et)(CO)(3)(py-NH(2))](PF(6)) (bpy-Et = 4-(N-(ethyl)aminocarbonyl)-4'-methyl-2,2'-bipyridine) (4-Et-NH(2)). The PEG complexes exhibited significantly higher water solubility and lower cytotoxicity (IC(50) = 6.6 to 1152 μM) than their PEG-free counterparts (IC(50) = 3.6 to 159 μM), indicating that the covalent attachment of a PEG pendant to rhenium(I) polypyridine complexes is an effective way to increase their biocompatibility. The amine complexes 1-PEG-NH(2)-4-PEG-NH(2) have been activated with thiophosgene to yield the isothiocyanate complexes [Re(N(∧)N)(CO)(3)(py-PEG-NCS)](PF(6)) (py-PEG-NCS = 3-isothiocyanato-5-(N-(2-(ω-methoxypoly(1-oxapropyl))ethyl)aminocarbonyl)pyridine; N(∧)N = phen (1-PEG-NCS), Me(4)-phen (2-PEG-NCS), Ph(2)-phen (3-PEG-NCS)), and [Re(bpy-PEG)(CO)(3)(py-NCS)](PF(6)) (py-NCS = 3-isothiocyanatopyridine) (4-PEG-NCS) as a new class of luminescent PEGylation reagents. To examine their PEGylation properties, these isothiocyanate complexes have been reacted with a model substrate n-butylamine, resulting in the formation of the thiourea complexes [Re(N(∧)N)(CO)(3)(py-PEG-Bu)](PF(6)) (py-PEG-Bu = 3-n-butylthioureidyl-5-(N-(2-(ω-methoxypoly(1-oxapropyl))ethyl)aminocarbonyl)pyridine; N(∧)N = phen (1-PEG-Bu), Me(4)-phen (2-PEG-Bu), Ph(2)-phen (3-PEG-Bu)), and [Re(bpy-PEG)(CO)(3)(py-Bu)](PF(6)) (py-Bu = 3-n-butylthioureidylpyridine) (4-PEG-Bu). Additionally, bovine serum albumin (BSA) and poly(ethyleneimine) (PEI) have been PEGylated with the isothiocyanate complexes to yield bioconjugates 1-PEG-BSA-4-PEG-BSA and 1-PEG-PEI-4-PEG-PEI, respectively. Upon irradiation, all the PEGylated BSA and PEI conjugates exhibited intense and long-lived emission in aqueous buffer under ambient conditions. The DNA-binding and polyplex-formation properties of conjugate 3-PEG-PEI have been studied and compared with those of unmodified PEI. Furthermore, the in vivo toxicity of complex 3-PEG-NH(2) and its PEG-free counterpart 3-Et-NH(2) has been investigated using zebrafish embryos as an animal model. Embryos treated with the PEG complex at high concentrations revealed delayed hatching, which has been ascribed to hypoxia as a result of adhering of the complex to the external surface of the chorion.

Spatial Emission Profiles for Flagging Matrix Interferences in Axial-Viewing Inductively Coupled Plasma-Atomic Emission Spectrometry: Part II - Statistical Protocol

A statistical protocol was developed and verified for automated signaling of matrix interferences in inductively coupled plasma-atomic emission spectrometry (ICP-AES). Spatial emission profiles in ICP-AES are versatile indicators for flagging matrix interference. A family of calibration curves is first generated by measurements of standard solutions at different spatial locations in the plasma. The determined-concentration profile of the analyte along a spatial measurement axis of the plasma is then obtained by analyzing the sample at each spatial location by reference to the respective calibration curve. The absence or presence of a matrix interference is gauged from the shape of the determined-concentration profile of the analyte. A flat determined-concentration profile indicates absence of matrix interference whereas a dissimilar (i.e., curved) concentration profile offers a clear warning signal that the analytical results are compromised by interferences. The developed protocol automatically classifies a spatial profile as flat or curved; it involves the computation of three statistical parameters: relative range0.05-0.95, σsample and σsuccessive. The term relative range0.05 0.95 refers to the ratio of the range to the mean of the relative-intensity (or determined concentration) values between the 5th and 95th percentiles in a spatial profile, whereas σsample and σsuccessive refer to the sample standard deviation and the standard deviation of successive values, respectively, of all values in a spatial profile. It was found that whenever the relative range0.05-0.95 of a spatial profile is below 1.5%, the profile can be considered to be flat and no further statistical testing is needed. If relative range0.05 0.95 > 1.5%, the σsuccessive / σsample ratio provides useful information on the flatness of the profile. If the profile is flat, σsuccessive will be statistically equivalent to σsample (i.e., σsuccessive / σsample = 1). In contrast, if curvature is present in the profile, σsuccessive will be statistically smaller than σsample (i.e., σsuccessive / σsample < 1). A statistical test, based on the transformation of the experimental σsuccessive / σsample ratio to the z-value of a standard normal distribution, was used to judge if the difference between σsuccessive and σsample is statistically significant. This statistical protocol for characterization of flatness in a spatial profile was verified in experiments carried out under the influence of various matrix interferences and different plasma operating conditions.

Biomimetic 3D Anisotropic Geometries by Uniaxial Stretch of Poly(ε-caprolactone) Films for Mesenchymal Stem Cell Proliferation, Alignment and Myogenic Differentiation

Anisotropic geometries are critical for eliciting cell alignment to dictate tissue microarchitectures and biological functions. Current fabrication techniques are complex and utilize toxic solvents, hampering their applications for translational research. Here, we present a novel simple, solvent-free and reproducible method via uniaxial stretching for incorporating anisotropic topographies on bioresorbable films with ambitions to realize stem cell alignment control. Uniaxial stretching of poly(ε-caprolactone) (PCL) films resulted in a 3D micro-ridge/groove topography (inter-ridge-distance: ~6 μm, ridge-length: ~90 μm, ridge-depth: 200-900 nm) with uniform distribution and controllable orientation by the direction of stretch on the whole film surface. Increasing stretch temperature (Ts) and draw ratio (DR), the inter-ridge-distance was reduced and ridge-length increased. Through modification of hydrolysis, increased surface hydrophilicity was achieved, while maintaining the morphology of PCL ridge/grooves. Upon seeding human mesenchymal stem cells (hMSCs) on uniaxial-stretched PCL films (UX-PCL), aligned hMSC organization was obtained. Compared to un-stretched films, hMSCs on UX-PCL had larger increase in cellular alignment (>85%) and elongation, without indication of cytotoxicity or reduction in cellular proliferation. This aligned hMSC organization was homogenous and stable-maintained with controlled orientation along the ridges on the whole UX-PCL surface for over two weeks. Moreover, the hMSCs on UX-PCL had a higher level of myogenic genes' expression than that on the un-stretched films. We conclude that uniaxial stretching has potential in patterning film topography with anisotropic structures. The UX-PCL in conjunction with hMSCs could be used as "basic units" to create tissue constructs with microscale control of cellular alignment and elongation for tissue engineering (T.E.) applications.

Common Cancer Risk and Statins: a Population-based Case-control Study in a Chinese Population

Objective: To investigate whether the use of statins is associated with common cancer risk. Methods: A population-based case-control study was conducted in Taiwan. Cases were defined as all patients who were aged 18 years and older and had received at least two statin prescriptions for use continuously for at least 6 months before a first-time diagnosis of studied cancers between the period of 2000 and 2008. The controls were matched to cases by age, sex, and index date. Adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) were estimated by using the Cox proportional hazards model. Results: A total of 6841 cases and 27,364 matched controls were analyzed. The adjusted hazard ratio for any statin use and cancer at any site were 0.76 (95% 0.654, 0.891). There were a significant reduced risk of gastric cancer (HR: 0.26, 95% CI: 0.107, 0.588), liver cancer (HR: 0.44, 95% CI: 0.279, 0.723) and uterine cancer (HR: 0.44, 95% CI: 0.279, 0.723) associated with any statins. Conclusion: Overall, the statins suggested a significant reduced risk of the most common cancers in a large Chinese population, particularly in gastric, liver, and uterine cancers.

Complement Component 3 Inhibition by an Antioxidant is Neuroprotective After Cerebral Ischemia and Reperfusion in Mice

Oxidative stress after stroke is associated with the inflammatory system activation in the brain. The complement cascade, especially the degradation products of complement component 3, is a key inflammatory mediator of cerebral ischemia. We have shown that proinflammatory complement component 3 is increased by oxidative stress after ischemic stroke in mice using DNA array. In this study, we investigated whether up-regulation of complement component 3 is directly related to oxidative stress after transient focal cerebral ischemia in mice and oxygen-glucose deprivation in brain cells. Persistent up-regulation of complement component 3 expression was reduced in copper/zinc-superoxide dismutase transgenic mice, and manganese-superoxide dismutase knockout mice showed highly increased complement component 3 levels after transient focal cerebral ischemia. Antioxidant N-tert-butyl-α-phenylnitrone treatment suppressed complement component 3 expression after transient focal cerebral ischemia. Accumulation of complement component 3 in neurons and microglia was decreased by N-tert-butyl-α-phenylnitrone, which reduced infarct volume and impaired neurological deficiency after cerebral ischemia and reperfusion in mice. Small interfering RNA specific for complement component 3 transfection showed a significant increase in brain cells viability after oxygen-glucose deprivation. Our study suggests that the neuroprotective effect of antioxidants through complement component 3 suppression is a new strategy for potential therapeutic approaches in stroke. © 2012 International Society for Neurochemistry, J. Neurochem. (2012) 10.1111/jnc.12111.

A New Class of Drug for the Management of Type 2 Diabetes: Sodium Glucose Co-transporter Inhibitors: 'Glucuretics'

Type 2 diabetes is a common, chronic disease with a prevalence that is increasing at epidemic proportions. Management involves advice on lifestyle changes, oral anti-hyperglycaemic agents and/or insulin. The kidneys play a major role in the regulation of glucose, re-absorbing 99% of the plasma glucose filtered through the renal glomeruli tubules. The glucose transporter, SGLT2, which is found primarily in the S1 segment of the proximal renal tubule accounts for 90% of glucose re-absorption. Competitive inhibition of SGLT2 induces glucosuria in a dose dependent manner and appears to have beneficial effects on glucose regulation in individuals with type 2 diabetes. O-glucoside phlorozin is the model substance for SGLT2 inhibitors: various O-, C-, N- and S-glucosides with varying affinity and specificity have been synthesised.

Chromatin Decondensation and Nuclear Softening Accompany Nanog Downregulation in Embryonic Stem Cells

The interplay between epigenetic modification and chromatin compaction is implicated in the regulation of gene expression, and it comprises one of the most fascinating frontiers in cell biology. Although a complete picture is still lacking, it is generally accepted that the differentiation of embryonic stem (ES) cells is accompanied by a selective condensation into heterochromatin with concomitant gene silencing, leaving access only to lineage-specific genes in the euchromatin. ES cells have been reported to have less condensed chromatin, as they are capable of differentiating into any cell type. However, pluripotency itself-even prior to differentiation-is a split state comprising a naïve state and a state in which ES cells prime for differentiation. Here, we show that naïve ES cells decondense their chromatin in the course of downregulating the pluripotency marker Nanog before they initiate lineage commitment. We used fluorescence recovery after photobleaching, and histone modification analysis paired with a novel, to our knowledge, optical stretching method, to show that ES cells in the naïve state have a significantly stiffer nucleus that is coupled to a globally more condensed chromatin state. We link this biophysical phenotype to coinciding epigenetic differences, including histone methylation, and show a strong correlation of chromatin condensation and nuclear stiffness with the expression of Nanog. Besides having implications for transcriptional regulation and embryonic cell sorting and suggesting a putative mechanosensing mechanism, the physical differences point to a system-level regulatory role of chromatin in maintaining pluripotency in embryonic development.

Clinical Impact of Drug-Drug Interaction Between Aspirin and Prednisolone at a Cancer Center

BACKGROUND: Adverse gastrointestinal (GI) events are complications in aspirin and prednisolone cotherapy. The prevalence of adverse GI events would be expected to be increased with cotherapy due to the overlapping toxicities of the 2 drugs. However, there is a dearth of literature investigating how often this interaction causes clinically important adverse GI events. OBJECTIVES: This retrospective study aimed to determine the prevalence of adverse GI events associated with the coadministration of aspirin and prednisolone. The use of gastroprotectant agents was also studied. METHODS: The medical records of patients with cancer prescribed aspirin and prednisolone therapy between January 2007 and June 2011 were analyzed. The duration of aspirin-prednisolone overlap, prevalence of adverse GI events, and details on the concurrent use of other medications were evaluated. RESULTS: The study included data from 142 patients (male, 64.8%; mean [SD] age, 67.4 [11.0] years). A total of 78.9% of the patients were on some form of gastroprotectant, the most common class of which was proton pump inhibitors. The prevalence of adverse GI events was 4.2% (6 patients). Four patients had presented with GI symptoms (abdominal pain, diarrhea, dysphagia, and vomiting); 3 patients had signs of GI injury (duodenal ulcers, iron deficiency anemia, and a Mallory-Weiss tear). The Naranjo algorithm classified 5 patients experienced possible adverse drug reactions (ADRs), and 1 as a probable ADR. CONCLUSION: Our study found that the prevalence of adverse GI events was low and managed to establish a weak association between the occurrence of events and the coadministration of aspirin and prednisolone. This finding, together with the concurrent prescription of gastroprotectants, suggests that the clinical impact of the aspirin and prednisolone DDI is minimal.

Intravenous Immunoglobulin Replacement Therapy to Prevent Pulmonary Infection in a Patient with Good's Syndrome

Good's syndrome is an acquired immunodeficiency state associated with thymoma and characterized by recurrent pulmonary infections. We describe a 67-year-old woman who presented with respiratory symptoms caused by concomitant disseminated cytomegalovirus infection and Pneumocystis jiroveci pneumonia 38 months after thymectomy for a thymoma. Immunologic analysis revealed hypogammaglobulinemia with absent B-cell population as demonstrated by flow cytometry, consistent with Good's syndrome. Following treatment with sulfamethoxazole/trimethoprim and ganciclovir, the patient improved with resolution of her respiratory symptoms. However, the patient subsequently experienced additional infections, necessitating additional subsequent hospital admissions. During the last admission, intravenous immunoglobulin (IVIG) replacement therapy was initiated and continued after discharge. Infection has been prevented for one year after beginning IVIG replacement therapy. This case reveals that in patients with combined humoral and cell-mediated immune deficiency, concomitant infection with different pathogens is not unusual, and immediate specific therapy is important. Periodic IVIG infusion, to maintain adequate Ig levels, is recommended.

Selective Induction of Glutathione S-transferases in Round Spermatids from the Brown-Norway Rat by the Chemotherapeutic Regimen for Testicular Cancer

Chemotherapeutic drugs can affect DNA in male germ cells, thereby impacting on the integrity of the genome transmitted to offspring. Drug metabolizing enzymes can protect cells from xenobiotic insult. We analyzed the expression pattern of such enzymes in isolated round spermatids from rats exposed to drugs used to treat testicular cancer: bleomycin, etoposide, and cis-platinum (BEP). The number of isozymes expressed and the overall relative expression values were highest for the glutathione S-transferases (GSTs). Moreover, BEP treatment significantly increased the expression of 8 GSTs and 3 aldehyde dehydrogenases. Increased expression of GST isozymes was confirmed by qRT-PCR and western blot analysis. Although Gst genes can be targets for epigenetic modifications, promoter DNA methylation was not affected by BEP treatment. As GSTs are involved in drug resistance mechanisms, we hypothesize that BEP induction of GST expression may lead to the survival of damaged germ cells and the production of abnormal sperm.

Measurement Uncertainty for Serum Free Light Chain Assays: Estimation and Implication on Result Interpretation

OBJECTIVE: To estimate the measurement uncertainty (MU) and reference change value (RCV) for serum free light chain assays (sFLC), and to review their implications on result interpretation. METHOD: Data from 6-9months of internal QC and up to 3.5years of EQA results were collected retrospectively, on the Roche Modular P analyzer and Dade Behring BN II nephelometer from two independent laboratories. MU was estimated from its components related to imprecision and bias determinations, while RCV was calculated from the estimated MU and published values on biological variation. RESULTS: The standard uncertainty related to imprecision for the FLC were similar between two instruments, ranging from ~4% to ~8%, so were the uncertainty related to bias, ranging between 24% and 44%. The overall MU with 95% coverage for Fκ, Fλ and their ratio (rFLC) were 55%, 87% and 103% (Modular P), and 49%, 76% and 91% (BNII) respectively. The RCVs with biological variation for Fκ, Fλ and rFLC were comparable between two instruments and averaged 106%, 138% and 173% respectively. DISCUSSIONS: The relatively method-independent MU reflected the sFLC limitations. For monoclonal gammopathy detection, the universal rFLC reference limits±MU gave an effective range of 0.02-3.15 (BNII) (as compared to 0.26-1.65), revealing substantial diagnostic grey zones on both ends. When monitoring disease activity, a minimum change (RCV) of ~100% in FLC concentrations or ~170% in rFLC would be considered significant. Applying MU when interpreting sFLC results puts current diagnostic cut-offs into perspective, and highlights the importance of collating other clinical findings.

Association Between Sex Hormones and Colorectal Cancer Risk in Men and Women

BACKGROUND & AIMS: There is observational and clinical evidence that indicate that sex hormones affect development of colorectal cancer (CRC) in men and women. However, the relationship between endogenous sex hormone levels and CRC is unclear. METHODS: We collected data on lifestyle, medical history, and diet etc. (through 2008), along with blood samples, from the Nurses' Health Study, the Women's Health Study, the Health Professional Follow-Up Study, and the Physicians' Health Study II. We measured plasma levels of estrone, estradiol, testosterone, sex hormone binding globulin (SHBG), and c-peptide among 730 women (293 cases of CRC and 437 healthy individuals, as controls) and 1158 men (439 CRC cases and 719 controls), and used unconditional logistic regression to estimate relative risks (RRs) and 95% confidence intervals (CIs). All statistical tests were 2-sided. RESULTS: Total testosterone, SHBG, and the ratio of estradiol to testosterone were associated with CRC in men after adjustments for matching and risk factors for CRC, including BMI and plasma levels of C-peptide. The RRs in the highest relative to the lowest quartile were 0.62 for testosterone (95% CI, 0.40-0.96), 0.65 for SHBG (95% CI, 0.42-0.99), and 2.63 for the ratio (95% CI, 1.58-4.36) (P-values for trend ≤0.02). However, in women, only the ratio of estradiol to testosterone was (inversely) associated with CRC after adjustments for all factors (RR, 0.43; 95% CI, 0.22-0.84; P-value for trend, .03). CONCLUSIONS: Based on combined data from 4 population studies, there appears to be an association between levels of sex hormones and CRC risk in men. There also appears to be an inverse association between the ratio of estradiol to testosterone and CRC in postmenopausal women.

Prostratin Exhibits Both Replication Enhancing and Inhibiting Effects on FIV Infection of Feline CD4(+) T-cells

The phorbol ester Prostratin may either stimulate or inhibit human immunodeficiency virus-1 (HIV-1) replication. Here we report that Prostratin also exhibits a similar dual action upon feline immunodeficiency virus (FIV) replication in an IL-2-dependent feline CD4+ T-cell line (MYA-1). While withdrawal of IL-2 halted FIV spread, Prostratin rescued virus production and cell viability, mimicking the functions of the cytokine. Conversely, FIV grew rapidly in the presence of IL-2 and this was inhibited by Prostratin. In contrast to HIV-1, Prostratin mediated inhibition of FIV through means other than blocking virus entry. Co-application of the protein kinase C (PKC) inhibitor Gö6850 with Prostratin reversed both the inhibitory and stimulatory effects, suggesting that PKC is crucial for FIV replication.

Accurate Measurement of Peripheral Blood Mononuclear Cell Concentration Using Image Cytometry to Eliminate RBC-Induced Counting Error

Peripheral blood mononuclear cells (PBMCs) have been widely researched in the fields of immunology, infectious disease, oncology, transplantation, hematological malignancy, and vaccine development. Specifically, in immunology research, PBMCs have been utilized to monitor concentration, viability, proliferation, and cytokine production from immune cells, which are critical for both clinical trials and biomedical research. The viability and concentration of isolated PBMCs are traditionally measured by manual counting with trypan blue using a hemacytometer. One of the common issues of PBMC isolation is red blood cell (RBC) contamination. The RBC contamination can be dependent on the donor sample and/or technical skill level of the operator. RBC contamination in a PBMC sample can introduce error to the measured concentration, which can pass down to future experimental assays performed on these cells. To resolve this issue, RBC lysing protocol can be used to eliminate potential error caused by RBC contamination. In the recent years, a rapid fluorescence-based image cytometry system has been utilized for bright-field and fluorescence imaging analysis of cellular characteristics (Nexcelom Bioscience LLC, Lawrence, MA). The Cellometer image cytometry system has demonstrated the capability of automated concentration and viability detection in disposable counting chambers of unpurified mouse splenocytes and PBMCs stained with acridine orange and propidium iodide under fluorescence detection. In this work, we demonstrate the ability of Cellometer image cytometry system to accurately measure PBMC concentration, despite RBC contamination, by comparison of five different total PBMC counting methods: (1) manual counting of trypan blue-stained PBMCs in hemacytometer, (2) manual counting of PBMCs in bright-field images, (3) manual counting of acetic acid lysing of RBCs with TB-stained PBMCs, (4) automated counting of acetic acid lysing of RBCs with PI-stained PBMCs, and (5) AO/PI dual staining method. The results show comparable total PBMC counting among all five methods, which validate the AO/PI staining method for PBMC measurement in the image cytometry method.

Chikungunya Infection in Malaysia: Comparison with Dengue Infection in Adults and Predictors of Persistent Arthralgia

BACKGROUND: Chikungunya virus (CHIKV) and dengue virus (DENV) co-circulate in areas endemic with the Aedes mosquito vectors. Both viruses cause similar illnesses which may be difficult to distinguish clinically. CHIKV is also associated with persistent arthralgia. OBJECTIVES: To compare and describe factors which differentiate between DENV and CHIKV infections on presentation; and to describe predictors of persistent arthralgia in CHIKV patients. STUDY DESIGN: Patients aged >14 years diagnosed with acute CHIKV and DENV infections in Kuala Lumpur, Malaysia were retrospectively identified. Clinical and laboratory data were obtained from medical records, and compared. CHIKV patients were telephoned 15-24 months later and interviewed about persistent symptoms. Logistic regression analysis was performed. RESULTS: A total of 53 CHIKV and 113 DENV patients were included. CHIKV patients were older and more likely to be female. CHIKV was independently associated with arthralgia and rash, while DENV was associated with myalgia, raised aspartate transaminase, and leucopaenia. Forty CHIKV patients were followed up, with a median duration of self-reported arthralgia of 3 months (range, 0-24 months). Eighteen (45%) had persistent arthralgia beyond 4 months, for which age >40 years was an independent predictor. At 1 year, 9 (22.5%) patients had arthralgia. CONCLUSION: In Kuala Lumpur, selected clinical and laboratory predictors help to distinguish between DENV and CHIKV infections. Persistent arthralgia was a frequent sequel of CHIKV infection in this cohort.

A "pre-seasonal" Hospital Outbreak of Influenza Pneumonia Caused by the Drift Variant A/Victoria/361/2011-like H3N2 Viruses, Hong Kong, 2011

BACKGROUND: Beginning from late 2011 and early 2012, increasing circulation of antigenically drifted influenza A/Victoria/361/2011-like H3N2 viruses within genotype 3 of the A/Victoria/208/2009 clade have been reported in multiple European countries and elsewhere. Whether these emerging viruses are associated with increased disease severity is unclear. OBJECTIVES: To report the clinical and virological findings of a moderately severe hospital outbreak of A/Victoria/361/2011-like viruses that occurred in November 2011 in Hong Kong. STUDY DESIGN: Clinical and virological hospital outbreak investigation. RESULTS: The outbreak occurred in an adult psychiatric ward in November 2011, a time well before the usual local seasonal influenza winter peak. Altogether, 7 patients and 1 healthcare-worker were affected (mean age, 47 [range, 34-61] years). The attack rates among patients and healthcare-workers were 33% (7/21) and 7% (1/15), respectively. Pneumonia developed in 38% (3/8) of cases; none had underlying immunocompromised conditions. High nasopharyngeal viral loads were detected. All cases responded to antiviral treatment. Multiple amino acid mutations with reference to earlier A(H3N2) vaccine strains were mapped to key antigenic sites on hemagglutinin; however, no critical mutations on receptor binding sites were detected. Viral sequence variations jeopardized the performance of molecular diagnostic assays. CONCLUSIONS: Severe disease and pneumonia occurred in a substantial proportion of non-immunocompromised adults in a hospital outbreak attributed to the emerging antigenically drifted A/Victoria/361/2011-like H3N2 viruses. Close monitoring of the transmission of this drift variant is required. Further studies are also necessary to determine virus virulence.

IL-15 and Macrophage Secretory Factors Facilitate Immune Activation of Neonatal Natural Killer Cells by Lipoteichoic Acid

Neonates possess a relatively "naive", yet inducible immune system. Our hypothesis is that upon strategic antigen exposure, cytokine priming and sensitization by accessory cells, natural killer (NK) cells could be activated to become a functional phenotype. We investigated the in vitro stimulation of cord blood (CB) and adult NK cells upon challenge with lipoteichoic acid (LTA), interleukin (IL)-15 and LTA-primed autologous macrophage-conditioned medium, using CD107a and CD69 phenotypes as indicators of activation. We also examined response of CB macrophages to LTA, in terms of P44/42 extracellular signal-regulated kinases (ERK1/2) activation and cytokine secretion. LTA significantly induced secretion of inflammatory cytokines tumor necrotic factor (TNF)-α, IL-6, IL-12 and activated the upstream signal of ERK1/2 phosphorylation in neonatal macrophages. The magnitude of responses to stimulation differed between neonatal and adult NK cells. Co-stimulation with IL-15 was critical for expansion of the CD69 and CD107a NK subpopulations in both neonatal and adult cells, upon a LTA challenge. NK cell activation could be enhanced by LTA-primed autologous macrophages through secretory factors. Our results indicated that neonatal macrophages and NK cells can evoke immunologic responses to a Gram-positive bacterial antigen. The combinatory priming strategy is relevant for development of novel protocols, such as IL-15 treatment, to compensate for the immaturity of the innate immune system in newborns against bacterial infections.

Modeling the Impact of Public Access Defibrillator Range on Public Location Cardiac Arrest Coverage

BACKGROUND: Public access defibrillation with automated external defibrillators (AEDs) can improve survival from out-of-hospital cardiac arrests (OHCA) occurring in public. Increasing the effective range of AEDs may improve coverage for public location OHCAs. OBJECTIVE: To quantify the relationship between AED effective range and public location cardiac arrest coverage. METHODS: This was a retrospective cohort study using the Resuscitation Outcomes Consortium Epistry database. We included all public-location, atraumatic, EMS-attended OHCAs in Toronto, Canada between Dec. 16, 2005 and July 15, 2010. We ran a mathematical model for AED placement that maximizes coverage of historical public OHCAs given pre-specified values of AED effective range and the number of locations to place AEDs. Locations of all non-residential buildings were obtained from the City of Toronto and used as candidate sites for AED placement. Coverage was evaluated for range values from 10 to 300 meters and number of AED locations from 10 to 200, both in increments of 10, for a total of 600 unique scenarios. Coverage from placing AEDs in all public buildings was also measured. Results: There were 1,310 public location OHCAs during the study period, with 25,851 non-residential buildings identified as candidate sites for AED placement. Cardiac arrest coverage increased with AED effective range, with improvements in coverage diminishing at higher ranges. For example, for a deployment of 200 AED locations, increasing effective range from 100m to 200m covered an additional 15% of cardiac arrests, whereas increasing range further from 200m to 300m covered an additional 10%. Placing an AED in each of the 25,851 public buildings resulted in coverage of 50% and 95% under assumed effective ranges of 50m and 300m, respectively. CONCLUSION: Increasing AED effective range can improve cardiac arrest coverage. Mathematical models can help evaluate the potential impact of initiatives which increase AED range.

Addendum: The Cancer Cell Line Encyclopedia Enables Predictive Modelling of Anticancer Drug Sensitivity

Point-spread Function Reconstruction in Ground-based Astronomy by L1-lp Model

In ground-based astronomy, images of objects in outer space are acquired via ground-based telescopes. However, the imaging system is generally interfered by atmospheric turbulence, and hence images so acquired are blurred with unknown point-spread function (PSF). To restore the observed images, the wavefront of light at the telescope's aperture is utilized to derive the PSF. A model with the Tikhonov regularization has been proposed to find the high-resolution phase gradients by solving a least-squares system. Here we propose the l1-lp (p=1, 2) model for reconstructing the phase gradients. This model can provide sharper edges in the gradients while removing noise. The minimization models can easily be solved by the Douglas-Rachford alternating direction method of a multiplier, and the convergence rate is readily established. Numerical results are given to illustrate that the model can give better phase gradients and hence a more accurate PSF. As a result, the restored images are much more accurate when compared to the traditional Tikhonov regularization model.

Long Chain N-acyl Homoserine Lactone Production by Enterobacter Sp. Isolated from Human Tongue Surfaces

We report the isolation of N-acyl homoserine lactone-producing Enterobacter sp. isolate T1-1 from the posterior dorsal surfaces of the tongue of a healthy individual. Spent supernatants extract from Enterobacter sp. isolate T1-1 activated the biosensor Agrobacterium tumefaciens NTL4(pZLR4), suggesting production of long chain AHLs by these isolates. High resolution mass spectrometry analysis of these extracts confirmed that Enterobacter sp. isolate T1-1 produced a long chain N-acyl homoserine lactone, namely N-dodecanoyl-homoserine lactone (C12-HSL). To the best of our knowledge, this is the first isolation of Enterobacter sp., strain T1-1 from the posterior dorsal surface of the human tongue and N-acyl homoserine lactones production by this bacterium.

Estimation of the Whole-body Averaged SAR of Grounded Human Models for Plane Wave Exposure at Respective Resonance Frequencies

According to the international guidelines, the whole-body averaged specific absorption rate (WBA-SAR) is used as a metric of basic restriction for radio-frequency whole-body exposure. It is well known that the WBA-SAR largely depends on the frequency of the incident wave for a given incident power density. The frequency at which the WBA-SAR becomes maximal is called the 'resonance frequency'. Our previous study proposed a scheme for estimating the WBA-SAR at this resonance frequency based on an analogy between the power absorption characteristic of human models in free space and that of a dipole antenna. However, a scheme for estimating the WBA-SAR in a grounded human has not been discussed sufficiently, even though the WBA-SAR in a grounded human is larger than that in an ungrounded human. In this study, with the use of the finite-difference time-domain method, the grounded condition is confirmed to be the worst-case exposure for human body models in a standing posture. Then, WBA-SARs in grounded human models are calculated at their respective resonant frequencies. A formula for estimating the WBA-SAR of a human standing on the ground is proposed based on an analogy with a quarter-wavelength monopole antenna. First, homogenized human body models are shown to provide the conservative WBA-SAR as compared with anatomically based models. Based on the formula proposed here, the WBA-SARs in grounded human models are approximately 10% larger than those in free space. The variability of the WBA-SAR was shown to be ±30% even for humans of the same age, which is caused by the body shape.

Autophagy Deficiency Leads to Protection from Obesity and Insulin Resistance by Inducing Fgf21 As a Mitokine

Despite growing interest and a recent surge in papers, the role of autophagy in glucose and lipid metabolism is unclear. We produced mice with skeletal muscle-specific deletion of Atg7 (encoding autophagy-related 7). Unexpectedly, these mice showed decreased fat mass and were protected from diet-induced obesity and insulin resistance; this phenotype was accompanied by increased fatty acid oxidation and browning of white adipose tissue (WAT) owing to induction of fibroblast growth factor 21 (Fgf21). Mitochondrial dysfunction induced by autophagy deficiency increased Fgf21 expression through induction of Atf4, a master regulator of the integrated stress response. Mitochondrial respiratory chain inhibitors also induced Fgf21 in an Atf4-dependent manner. We also observed induction of Fgf21, resistance to diet-induced obesity and amelioration of insulin resistance in mice with autophagy deficiency in the liver, another insulin target tissue. These findings suggest that autophagy deficiency and subsequent mitochondrial dysfunction promote Fgf21 expression, a hormone we consequently term a 'mitokine', and together these processes promote protection from diet-induced obesity and insulin resistance.

Assessments of the Effects of Nicotine and Ketamine Using Tyrosine Hydroxylase-green Fluorescent Protein Transgenic Zebrafish As Biosensors

Transgenic zebrafish are a common vertebrate model system for the study of addictive behavior. In the present study, plasmid constructs containing green fluorescent protein (GFP) and the promoter of tyrosine hydroxylase (TH), a key synthetic enzyme for catecholamines, were produced. The TH-GFP constructs were microinjected into zebrafish embryonic cells. Three days post-fertilization, GFP began expressing in distinct catecholaminergic areas. The TH-GFP transgenic zebrafish were employed as live biosensors to test the effects of the commonly abused drugs nicotine and ketamine. First, locomotion assays were used to study the general excitatory effects of the drugs. Maximal locomotor activity was obtained after treatment with a high concentration of nicotine (10μM), but with a much lower concentration of ketamine (0.1μM). Second, TH protein levels in zebrafish brains were assessed by Western blot. TH protein levels were significantly increased, with maximal protein levels found after treatment with the same drug concentrations that gave maximal locomotor activity. Importantly, analysis of GFP in the zebrafish catecholaminergic areas revealed the same expression patterns as was obtained by Western blot. The present results indicate that increased locomotor activity can be correlated to TH protein expression, as indicated by Western blot and expression of TH-GFP. We have shown that TH-GFP expression is a reliable method to show the effects of drugs on TH expression that may be employed as a novel high-throughput live biosensor for screening drugs of abuse.

Enhancement of Commercial Antifungal Agents by Kojic Acid

Natural compounds that pose no significant medical or environmental side effects are potential sources of antifungal agents, either in their nascent form or as structural backbones for more effective derivatives. Kojic acid (KA) is one such compound. It is a natural by-product of fungal fermentation commonly employed by food and cosmetic industries. We show that KA greatly lowers minimum inhibitory (MIC) or fungicidal (MFC) concentrations of commercial medicinal and agricultural antifungal agents, amphotericin B (AMB) and strobilurin, respectively, against pathogenic yeasts and filamentous fungi. Assays using two mitogen-activated protein kinase (MAPK) mutants, i.e., sakA∆, mpkC∆, of Aspergillus fumigatus, an agent for human invasive aspergillosis, with hydrogen peroxide (H(2)O(2)) or AMB indicate such chemosensitizing activity of KA is most conceivably through disruption of fungal antioxidation systems. KA could be developed as a chemosensitizer to enhance efficacy of certain conventional antifungal drugs or fungicides.

Injurious Effects of Emodin on Maturation of Mouse Oocytes, Fertilization and Fetal Development Via Apoptosis

Emodin (1,3,8-trihydroxy-6-methylanthraquinone), a major constituent of rhubarb, has a wide range of therapeutic applications. Previous studies have established that emodin induces apoptosis in the inner cell mass and trophectoderm of mouse blastocysts and leads to decreased embryonic development and viability, indicating a role as an injury risk factor for normal embryonic development. However, the mechanisms underlying its hazardous effects have yet to be characterized. In the current study, we further investigated the effects of emodin on oocyte maturation and subsequent pre- and post-implantation development, both in vitro and in vivo. Notably, emodin induced a significant reduction in the rates of oocyte maturation, fertilization, and in vitro embryonic development. Treatment of oocytes with emodin during in vitro maturation (IVM) led to increased resorption of postimplantation embryos and decreased fetal weight. Experiments using an in vivo mouse model disclosed that consumption of drinking water containing 20-40 μM emodin led to decreased oocyte maturation and in vitro fertilization, as well as early embryonic developmental injury. Notably, pretreatment with a caspase-3-specific inhibitor effectively prevented emodin-triggered injury effects, suggesting that impairment of embryo development occurs via a caspase-dependent apoptotic process.

Predicting Retention Times of Naturally Occurring Phenolic Compounds in Reversed-Phase Liquid Chromatography: A Quantitative Structure-Retention Relationship (QSRR) Approach

Quantitative structure-retention relationships (QSRRs) have successfully been developed for naturally occurring phenolic compounds in a reversed-phase liquid chromatographic (RPLC) system. A total of 1519 descriptors were calculated from the optimized structures of the molecules using MOPAC2009 and DRAGON softwares. The data set of 39 molecules was divided into training and external validation sets. For feature selection and mapping we used step-wise multiple linear regression (SMLR), unsupervised forward selection followed by step-wise multiple linear regression (UFS-SMLR) and artificial neural networks (ANN). Stable and robust models with significant predictive abilities in terms of validation statistics were obtained with negation of any chance correlation. ANN models were found better than remaining two approaches. HNar, IDM, Mp, GATS2v, DISP and 3D-MoRSE (signals 22, 28 and 32) descriptors based on van der Waals volume, electronegativity, mass and polarizability, at atomic level, were found to have significant effects on the retention times. The possible implications of these descriptors in RPLC have been discussed. All the models are proven to be quite able to predict the retention times of phenolic compounds and have shown remarkable validation, robustness, stability and predictive performance.

Transitioning Inpatients to Housing: a Group Discharge Approach

Development and Validation of an MRI Reference Criterion for Defining a Positive SIJ MRI in Spondyloarthritis

OBJECTIVE: To validate an MRI reference criterion for a positive SIJ MRI based on the level of confidence in classification of spondyloarthritis (SpA) by expert MRI readers. METHODS: Four readers assessed SIJ MRI in two inception cohorts (A/B) of 157 consecutive back pain patients ≤50 years, and in 20 healthy controls. Patients were classified according to clinical examination and pelvic radiography as having non-radiographic axial SpA (n=51), ankylosing spondylitis (n=34), or non-specific back pain (n=72). Readers recorded their level of confidence in the classification of SpA on a 0-10 scale (0=definitely not; 10=definite). The MRI reference criterion was pre-specified as the majority of readers recording a confidence of 8-10; absence of SpA required all readers to record Non-SpA (confidence 0-4). We calculated inter-reader reliability and agreement between MRI-based and clinical classification using kappa statistics. We estimated cut-off values for MRI lesions attaining specificity `0.90 for SpA. RESULTS: 76.4%/71.6% of subjects in cohorts A/B met the MRI criterion. Kappa values for inter-reader agreement were 0.76/0.80, and between MRI-based and clinical assessment 0.93/0.57. Using this MRI reference criterion, the cut-off for number of affected SIJ quadrants needed to reach a pre-defined specificity `0.90 was `2/`2 for BME and `1/`1 for erosion in cohorts A/B, and both lesions BME and/or erosion increased sensitivity without reducing specificity. CONCLUSION: This data-driven study using two inception cohorts and comparing clinical and MRI-based classification supports the case for including both erosion and BME to define a positive SIJ MRI for the classification of axial SpA. © 2012 by the American College of Rheumatology.

1-(2'-Anilinyl)prop-2-yn-1-ol Rearrangement for Oxindole Synthesis

A synthetic method that relies on NIS (N-iodosuccinimide)-mediated cycloisomerization reactions of 1-(2'-anilinyl)prop-2-yn-1-ols to gem-3-(diiodomethyl)indolin-2-ones and 2-(iodomethylene)indolin-3-ones has been developed. The reactions were shown to be chemoselective, with secondary and tertiary alcoholic substrates exclusively giving the 3- and 2-oxindole products, respectively. In the case of the latter, the transformation features an unprecedented double 1,2-OH and 1,2-alkyl migration relay. Density functional theory (DFT) calculations based on proposed iodoaminocyclization species provide insight into this unique divergence in product selectivity.

Toxin-Antitoxin Genes of the Gram-Positive Pathogen Streptococcus Pneumoniae: So Few and Yet So Many

Summary: Pneumococcal infections cause up to 2 million deaths annually and raise a large economic burden and thus constitute an important threat to mankind. Because of the increase in the antibiotic resistance of Streptococcus pneumoniae clinical isolates, there is an urgent need to find new antimicrobial approaches to triumph over pneumococcal infections. Toxin-antitoxin (TA) systems (TAS), which are present in most living bacteria but not in eukaryotes, have been proposed as an effective strategy to combat bacterial infections. Type II TAS comprise a stable toxin and a labile antitoxin that form an innocuous TA complex under normal conditions. Under stress conditions, TA synthesis will be triggered, resulting in the degradation of the labile antitoxin and the release of the toxin protein, which would poison the host cells. The three functional chromosomal TAS from S. pneumoniae that have been studied as well as their molecular characteristics are discussed in detail in this review. Furthermore, a meticulous bioinformatics search has been performed for 48 pneumococcal genomes that are found in public databases, and more putative TAS, homologous to well-characterized ones, have been revealed. Strikingly, several unusual putative TAS, in terms of components and genetic organizations previously not envisaged, have been discovered and are further discussed. Previously, we reported a novel finding in which a unique pneumococcal DNA signature, the BOX element, affected the regulation of the pneumococcal yefM-yoeB TAS. This BOX element has also been found in some of the other pneumococcal TAS. In this review, we also discuss possible relationships between some of the pneumococcal TAS with pathogenicity, competence, biofilm formation, persistence, and an interesting phenomenon called bistability.

Functional Characterization of Reductive Dehalogenases Using Blue Native Polyacrylamide Gel Electrophoresis

Dehalococcoides mccartyi strains are obligate organohalide-respiring bacteria harboring multiple distinct reductive dehalogenase (RDase) genes within their genomes. A major challenge is to identify substrates for the enzymes encoded by these RDase genes. We demonstrate an approach that involves blue native polyacrylamide gel electrophoresis (BN-PAGE) followed by enzyme activity assays with gel slices and subsequent identification of proteins in gel slices using liquid chromatography tandem mass spectrometry (LC-MS/MS). RDase expression was investigated in cultures of Dehalococcoides mccartyi strain BAV1 and in the KB-1 consortium growing on chlorinated ethenes and 1,2-dichloroethane. In cultures of strain BAV1, BvcA was the only RDase detected, revealing that this enzyme catalyzes the dechlorination not only of vinyl chloride, but also of all dichloroethene isomers and 1,2-dichloroethane. In cultures of consortium KB-1, five distinct Dehalococcoides RDases and one Geobacter RDase were expressed under the conditions tested. Three of the five RDases included orthologs to the previously identified chlorinated ethene-dechlorinating enzymes VcrA, BvcA and TceA. This study revealed substrate promiscuity for these three enzymes, and provides a path forward to further explore the largely unknown RDase protein family.

BSPAR ANNUAL CONFERENCE ABSTRACTS * Oral Presentations * O1. The Impact of Modern Management on Outcomes of JIA Compared with Healthy Controls

Background: Development of modern therapeutic approaches over the last decade has improved short-term well-being in JIA patients. This study examines disease damage and body composition (BC). Aims: Comparison of JIA patients' clinical well-being with matched controls assessing BC, cross-sectional inflammatory markers and the levels of joint and uveitis-associated damage relating to their management. Methods: 112 JIA patients (disease duration ≥5 years) were cross-sectionally compared with 127 age, sex, ethnicity and deprivation score-matched controls, measuring inflammatory markers (hsCRP, IL-6 and ICAM-1), growth and BC [height, weight, fat mass (FM/H(2)), free fat mass (FFM/H(2)), BMI and skinfold thickness]. Clinical examination recorded disease damage, supported by case-note review. Results: Compared with controls, patients <16 years had indistinguishable values of hsCRP (P = 0.59) and ICAM-1 (P = 0.13), but > 16 years had higher hsCRP, ICAM-1 and IL-6 (P < 0.0001). Patients < 22 years had higher Z-BMI (P = 0.019), Z-waist circumference (P = 0.001) and FM/H(2) [using Tanita equations (P = 0.007), sum of four skinfolds (P = 0.005)], with 21.3% meeting obesity criteria (vs controls 5.6%) with no association with non-JIA risk factors for obesity. Compared with controls, all anthropometric equations revealed no difference in patients >22 years. Of 90 patients, Group A never required DMARDs, Group B had modern therapeutic approaches [prompt MTX (SSZ in 1) and rapid escalation to biologics] and Group C had late DMARDs/biologics intervention despite persistent inflammation. Feet, knees and temporomandibular joints were most commonly damaged. A positive correlation was found between delay in starting modern therapy and severest joint damage outcome (Table 1). Conclusions: It is of new concern that high obesity levels in the youngest patients were significantly greater than in controls. Normal inflammatory markers, reduced frequency and severity of joint damage but high complication rates of chronic anterior uveitis (CAU) were associated with modern therapeutic approaches. Disclosure statement: The authors have declared no conflict of interest. Table 1Damage to joints and eyes by disease management ManifestationA (n = 16)B (n = 59)C (n = 15)Joint damage    None14401    Radiological change only020    Minimal restriction293    Functional disability087    Joint replacement (hip and or knee)004    CAU ever285    Complications of CAU044    Cataract34    Glaucoma42    Permanent visual impairment21    Band keratopathy12    Posterior synechiae11.

Effects of Continuous Positive Airway Pressure on Neurocognitive Function in Obstructive Sleep Apnea Patients: The Apnea Positive Pressure Long-term Efficacy Study (APPLES)

To determine the neurocognitive effects of continuous positive airway pressure (CPAP) therapy on patients with obstructive sleep apnea (OSA).

Comparison of Consumer Perception and Acceptability for Steaks Cooked to Different Endpoints: Validation of Photographic Approach

Photographs have been used to enhance consumer reporting of preference of meat doneness, however, the use of photographs has not been validated for this purpose. This study used standard cooking methods to produce steaks of five different degrees of doneness (rare medium, medium well, well done and very well done) to study the consumer's perception of doneness, from both the external and internal surface of the cooked steak and also from corresponding photographs of each sample. Consumers evaluated each surface of the cooked steaks in relation to doneness for acceptability, 'just about right' and perception of doneness. Data were analysed using a split plot ANOVA and least significant test. Perception scores (for both external and internal surfaces) between different presentation methods (steak samples and corresponding photos), were not significantly different (p>0.05). The result indicates that photographs can be used as a valid approach for assessing preference for meat doneness.

The Emotional Side of Cognitive Distraction: Implications for Road Safety

Driver distraction is estimated to be one of the leading causes of motor vehicle accidents. However, little is known about the role of emotional distraction on driving, despite evidence that attention is highly biased toward emotion. In the present study, we used a dual-task paradigm to examine the potential for driver distraction from emotional information presented on roadside billboards. This purpose was achieved using a driving simulator and three different types of emotional information: neutral words, negative emotional words, and positive emotional words. Participants also responded to target words while driving and completed a surprise free recall task of all the words at the end of the study. The findings suggest that driving performance is differentially affected by the valence (negative versus positive) of the emotional content. Drivers had lower mean speeds when there were emotional words compared to neutral words, and this slowing effect lasted longer when there were positive words. This may be due to distraction effects on driving behavior, which are greater for positive arousing stimuli. Moreover, when required to process non-emotional target stimuli, drivers had faster mean speeds in conditions where the targets were interspersed with emotional words compared to neutral words, and again, these effects lasted longer when there were positive words. On the other hand, negative information led to better memory recall. These unique effects may be due to separate processes in the human attention system, particularly related to arousal mechanisms and their interaction with emotion. We conclude that distraction that is emotion-based can modulate attention and decision-making abilities and have adverse impacts on driving behavior for several reasons.

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