Metabolic reprogramming and Notch activity distinguish between non-small cell lung cancer subtypes.
Previous studies suggested that the metabolism is differently reprogrammed in the major subtypes of non-small cell lung cancer (NSCLC), squamous cell carcinomas (SCC) and adenocarcinomas (AdC). However, a comprehensive analysis of this differential metabolic reprogramming is lacking. Publicly available gene expression data from human lung cancer samples and cell lines were analysed. Stable isotope resolved metabolomics were performed on SCC and ADC tumours in human patients and in freshly resected tumour slices. Analysis of multiple transcriptomics data from human samples identified a SCC-distinguishing enzyme gene signature. SCC tumours from patients infused with [U-13C]-glucose and SCC tissue slices incubated with stable isotope tracers demonstrated differential glucose and glutamine catabolism compared to AdCs or non-cancerous lung, confirming increased activity through pathways defined by the SCC metabolic gene signature. Furthermore, the upregulation of Notch target genes was a distinguishing feature of SCCs, which correlated with the metabolic signature. Notch and MYC-driven murine lung tumours recapitulated the SCC-distinguishing metabolic reprogramming. However, the differences between SCCs and AdCs disappear in established cell lines in 2D culture. Our data emphasise the importance of studying lung cancer metabolism in vivo. They also highlight potential targets for therapeutic intervention in SCC patients including differentially expressed enzymes that catalyse reactions in glycolysis, glutamine catabolism, serine, nucleotide and glutathione biosynthesis.


EDTA-functionalized magnetic nanoparticles: A suitable platform for the analysis of low abundance urinary proteins.
Urine is a highly attractive source of biological information and disease biomarkers, whose proteome characterization is ongoing. To that end, depletion/enrichment strategies for protein analysis can be of great convenience. We have thus developed a method based on the use of EDTA-functionalized magnetic nanoparticles (NPs@EDTA), to fractionate urine samples before liquid chromatography-mass spectrometry analysis and compared the identified proteins with those obtained from ultrafiltrated/unfractionated (UF) urine samples. NPs@EDTA allowed larger urine volumes to be processed, resulting in a greater number of protein identifications (~2-fold) at a lower cost when compared to UF samples. Proteins of greater abundance (such as albumin and uromodulin) were, at least partially, depleted with NPs@EDTA while those of lower abundance were enriched. Bioinformatics analysis showed that approximately 27% of NPs@EDTA-enriched proteins were annotated as displaying enzymatic activity, most of these being hydrolytic enzymes (56%), particularly proteases/peptidases (48%). Also, post-translational modifications were prominently predicted across NPs@EDTA-enriched proteins (90%), particularly glycosylation (52%), phosphorylation (47%) and acetylation (30%). NPs@EDTA allowed the identification of 109 proteins in urine for the first time, showing high potential as a platform for urine's fractionation prior to proteomic analysis.


What is the role of the orthopaedic surgeon in management of fragility fractures?
Fragility fractures are the most prevalent trauma condition that orthopaedic surgeons face today. Osteoporosis and susceptibility to falls are the key predisposing factors. Despite evidence supporting the impact of treating osteoporosis on reducing the incidence of fragility fractures, it is often left untreated. Orthopaedic surgeons are often the first physicians to assess and treat the patient after a fragility fracture. Their role therefore does not end in the skillful fixation of the fractures, but they have a unique opportunity to ensure that preventive measures are implemented. This includes falls prevention, investigation of possible causes underlying osteoporosis, attention to diet, exercise, calcium, and vitamin D supplementation as well as prescription of anti-resorptive and anabolic medication. The need for a dedicated multidisciplinary team needs to be emphasized and therefore effective communication between the different parties is of paramount importance.


Expanding the clinical spectrum of the 'HDAC8-phenotype' - implications for molecular diagnostics, counseling and risk prediction.
Cornelia de Lange syndrome (CdLS) is a clinically heterogeneous disorder characterized by typical facial dysmorphism, cognitive impairment and multiple congenital anomalies. Approximately 75% of patients carry a variant in one of the five cohesin-related genes NIPBL, SMC1A, SMC3, RAD21 and HDAC8. Herein we report on the clinical and molecular characterization of 11 patients carrying 10 distinct variants in HDAC8. Given the high number of variants identified so far, we advise sequencing of HDAC8 as an indispensable part of the routine molecular diagnostic for patients with CdLS or CdLS-overlapping features. The phenotype of our patients is very broad, whereas males tend to be more severely affected than females, who instead often present with less canonical CdLS features. The extensive clinical variability observed in the heterozygous females might be at least partially associated with a completely skewed X-inactivation, observed in seven out of eight female patients. Our cohort also includes two affected siblings whose unaffected mother was found to be mosaic for the causative mutation inherited to both affected children. This further supports the urgent need for an integration of highly sensitive sequencing technology to allow an appropriate molecular diagnostic, genetic counseling and risk prediction.


Lessons from the network and next steps: the Wisconsin Head and Neck Cancer Network.
In summary, the Wisconsin Head and Neck Cancer Control Network was formulated with the primary charge of improving the management of patients with head and neck cancer throughout the State. Detailed data on over 1500 patients representing 80 percent of all head and neck cancer patients in the State of Wisconsin was gathered, reviewed for quality, and analyzed. The data base consisted of information on the symptoms, signs, physical findings, treatment, complications, morbidity, and outcome. Although it was the impression of Network members that lesions were being noted earlier and treatment perhaps more effective, it was not possible to document this impression with the available data. The benchmarks of success of the overall project were numerous, however, particularly in the areas of public and professional education. The individuals reached were extremely receptive to new information. Several patients indicated that their dentist and dental hygienists, for example, were performing more thorough oral examinations. The effort was successful in establishing a network of physicians and institutions throughout the State cooperating with a common interest. It is hoped that in the future we can continue to build on the experience of the Network. Follow-up data has continued to accumulate through a largely voluntary effort of the coinvestigators, and this will enhance the value of the data base. Such follow-up data will certainly facilitate ongoing and future research projects based on this comprehensive information. To date, one publication has been prepared since termination of the project using the data base.


Effect of early estrogen replacement therapy on microvascular reactivity in patients after bilateral ovarectomy.
The aim of the study was to evaluate skin microvascular reactivity (MVR) measured by laser Doppler flowmetry in women early after bilateral ovarectomy treated with oral and transdermal estrogen replacement therapy (ERT). Interventional, randomized study with a cross-over design. 41 patients (49+/-6 years, 6-12 weeks after surgical castration) were treated with 17-beta-estradiol transdermally (0.05 mg/day) or orally (2 mg/day) for three months and 20 healthy female subjects (47+/-5 years) served as controls. Records of laser Doppler flowmetry were blinded prior to the evaluation. Maximal perfusion and velocity of perfusion increase during post-occlusive reactive hyperemia (PORH) were lower before ERT comparing to controls at baseline (36+/-16 vs. 48+/-18 PU, p<0.05, and 2.8+/-1.9 vs. 4.2+/-2.3 PU, p<0.05, respectively). Velocity of perfusion increase in PORH decreased after oral ERT compared to baseline and also to transdermal ERT (2.1+/-1.2 vs. 2.8+/-1.9 PU.s-1, p<0.05, and vs. 3.5+/-3.2 PU.s-1, p<0.01, respectively), nonsignificant increase of this parameter after transdermal ERT led to normalization when comparing to control group (3.6+/-3.2 vs. 4.2+/-2.3 PU.s-1, NS). Increase of HDL-cholesterol and decrease of LDL-cholesterol (2.1+/-0.4 vs. 1.8+/-0.4 mmol.l-1, p<0.01, and 2.5+/-0.7 vs. 3.1+/-1.0 mmol.l-1, p<0.01) was observed after oral ERT while HDL-cholesterol increase after transdermal ERT was less pronounced (1.96+/-0.42 mmol.l-1, p<0.05). LDL-cholesterol levels did not change. A correlation between HDL-cholesterol and maximal post-occlusive flow expressed in % of basal perfusion was observed in patients before treatment (r=0.47, p=0.002). Microvascular reactivity is impaired in women early after bilateral ovarectomy. No statistically significant improvement of MVR was observed after oral estrogen replacement therapy, normalization of MVR after transdermal ERT was only partial. Changes of MVR and lipid profile differed between oral and transdermal routes of estrogen replacement therapy.


Color Stability Assessment of Two Different Composite Resins with Variable Immersion Time Using Various Beverages: An In vitro Study.
The aim of the study was to evaluate the difference in the color of microhybrid (MH) and nanofilled (NF) composite resins after 24 and 48 h in beverages such as red wine (RW), Coca-Cola, and distilled water. The specific objective of this study was to investigate the cumulative effect of the colorant solutions on the dental composites. MH and NF composite resins (A2 shade) were used in this current study. Sixty disk-shaped material specimens (10 mm in diameter × 2 mm in thickness) were prepared using a fiber mold (ring), with the desired dimensions. The specimen surfaces were polished using super-snap polishing system. Sixty specimens were divided into two groups of 30 each (Group I: MH resin composite; Group II: NF resin composite). Both the groups divided into six subgroups (Subgroup I: RW for 24 h [RW-24]; Subgroup II: RW for 48 h; Subgroup III: Coca-Cola for 24 h [CC-24]; Subgroup IV: Coca-Cola for 48 h [CC-48]; Subgroup V: Distilled water for 24 h [DW-24]; Subgroup VI: Distilled water for 48 h [DW-48]). All the samples were immersed in respective drinks for a period of 24 h, and color differences were measured using ultraviolet spectrophotometer. Once again, all the samples were immersed for another 24 h in the same drinks. After 48 h, the color change of the samples was measured. Measurements were made according to the CIE L × a × b × color space relative to the CIE standard illuminant D65. The color changes of the specimens were evaluated using the following formula: Statistical analysis was performed. The data were analyzed using the one-way ANOVA and t-test at a significance level of 0.05. Color stability of MH composite resin was found to be inferior than the NF resin composite irrespective of immersion medium and time. In RW, the color change observed was maximum for both composite resins followed by Coca-Cola. Immersing the resin composites in distilled water for 24 and 48 h had negligible color change. A 48-h immersion of both composite resins in all three immersion mediums showed greater color change than 24 h immersion.


Healthier lifestyles after retirement in Europe? Evidence from SHARE.
This paper investigates changes in health behaviours upon retirement, using data drawn from the Survey of Health Ageing and Retirement in Europe. By exploiting changes in eligibility rules for early and statutory retirement, we identify the causal effect of retiring from work on smoking, alcohol drinking, engagement in physical activity and visits to the general practitioner or specialist. We provide evidence about individual heterogeneous effects related to gender, education, net wealth, early-life conditions and job characteristics. Our main results--obtained using fixed-effect two-stage least squares--show that changes in health behaviours occur upon retirement and may be a key mechanism through which the latter affects health. In particular, the probability of not practicing any physical activity decreases significantly after retirement, and this effect is stronger for individuals with higher education. We also find that different frameworks of European health care systems (i.e. countries with or without a gate-keeping system to regulate the access to specialist services) matter in shaping individuals' health behaviours after retirement. Our findings provide important information for the design of policies aiming to promote healthy lifestyles in later life, by identifying those who are potential target individuals and which factors may affect their behaviour. Our results also suggest the importance of policies promoting healthy lifestyles well before the end of the working life in order to anticipate the benefits deriving from individuals' health investments.


MC1R, ASIP, and DNA repair in sporadic and familial melanoma in a Mediterranean population.
Melanoma risk factors include fair pigmentation, multiple nevi, low DNA repair capacity, and CDKN2A or CDK4 mutations. Variants of the melanocortin-1 receptor (MC1R) gene have been associated with fair pigmentation and melanoma risk, and a polymorphism of the Agouti Signaling Protein (ASIP) gene has been associated with dark pigmentation. We examined MC1R and ASIP genotypes in relation to phenotypic characteristics, sporadic and familial melanoma risk, and melanoma thickness as an indicator of disease progression in a Mediterranean population. We studied 267 melanoma patients and 382 control subjects from a case-control study and a family study in northeastern Italy. Host factors were assessed by physical examination, questionnaire, spectrophotometer, and minimal erythema dose measurement. MC1R was sequenced, ASIP was genotyped, and DNA repair capacity was measured by the host-cell reactivation assay. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by logistic regression models. Effect modification of the association between MC1R and melanoma risk by phenotypic characteristics and DNA repair capacity was also assessed. All statistical tests were two-sided. Carrying MC1R variant alleles was associated with a two- to fourfold increase in risk of both sporadic and familial melanoma compared with carrying wild-type MC1R, particularly in individuals carrying multiple variant alleles (OR = 3.9; 95% CI = 3.3 to 4.6). This association was stronger in individuals with fewer additional risk factors (those with dark skin or few nevi). MC1R variant allele carriers were also three to four times more likely than were non-carriers to have thick melanomas. The ASIP polymorphism was not associated with pigmentation, nevi, or melanoma risk. MC1R was associated with melanoma risk and progression in a Mediterranean population, particularly in the absence of other strong risk factors, such as freckling or many nevi.


Baseline determination in social, health, and genetic areas in communities affected by glyphosate aerial spraying on the northeastern Ecuadorian border.
The northeastern Ecuadorian border has undergone aerial spraying with an herbicide mix that contains surfactants and adjuvants, executed by the Colombian Government. The purpose of this study was to diagnose social, health, and genetic aspects of the people affected by glyphosate. For this objective to be achieved, 144 people were interviewed, and 521 medical diagnoses and 182 peripheral blood samples were obtained. Genotyping of GSTP1 Ile105Val, GPX-1 Pro198Leu, and XRCC1 Arg399Gln polymorphisms were analyzed, using PCR-RFLP technique. The assessment of chromosomal aberrations was performed, obtaining 182 karyotypes. Malnutrition in children was 3%. Of the total population, 7.7% had children with malformations, and the percentage of abortions was 12.7%. Concerning genotyping, individuals with GSTP1 Val/Val obtained an odds ratio of 4.88 (p < 0.001), and Ile/Val individuals, together with Val/Val individuals, had an odds ratio of 2.6 (p < 0.05). In addition, GPX-1 Leu/Leu individuals presented an odds ratio (OR) of 8.5 (p < 0.05). Regarding karyotyping, the 182 individuals had normal karyotypes. In conclusion, the study population did not present significant chromosomal and DNA alterations. The most important social impact was fear. We recommend future prospective studies to assess the communities.


A newly synthesized sinapic acid derivative inhibits endothelial activation in vitro and in vivo.
Inhibition of oxidative stress and inflammation in vascular endothelial cells (ECs) may represent a new therapeutic strategy against endothelial activation. Sinapic acid (SA), a phenylpropanoid compound, is found in natural herbs and high-bran cereals and has moderate antioxidant activity. We aimed to develop new SA agents with the properties of antioxidation and blocking EC activation for possible therapy of cardiovascular disease. We designed and synthesized 10 SA derivatives according to their chemical structures. Preliminary screening of the compounds involved scavenging hydroxyl radicals and 2,2-diphenyl-1-picrylhydrazyl (DPPH(⋅)), croton oil-induced ear edema in mice, and analysis of the mRNA expression of adhesion molecules in ECs. 1-Acetyl-sinapic acyl-4-(3'-chlorine-)benzylpiperazine (SA9) had the strongest antioxidant and anti-inflammatory activities both in vitro and in vivo. Thus, the effect of SA9 was further studied. SA9 inhibited tumor necrosis factor α-induced upregulation of adhesion molecules in ECs at both mRNA and protein levels, as well as the consequent monocyte adhesion to ECs. In vivo, result of face-to-face immunostaining showed that SA9 reduced lipopolysaccharide-induced expression of intercellular adhesion molecule-1 in mouse aortic intima. To study the molecular mechanism, results from luciferase assay, nuclear translocation of NF-κB, and Western blot indicated that the mechanism of the anti-inflammatory effects of SA9 might be suppression of intracellular generation of ROS and inhibition of NF-κB activation in ECs. SA9 is a prototype of a novel class of antioxidant with anti-inflammatory effects in ECs. It may represent a new therapeutic approach for preventing endothelial activation in cardiovascular disorders.


Fast and accurate automatic structure prediction with HHpred.
Automated protein structure prediction is becoming a mainstream tool for biological research. This has been fueled by steady improvements of publicly available automated servers over the last decade, in particular their ability to build good homology models for an increasing number of targets by reliably detecting and aligning more and more remotely homologous templates. Here, we describe the three fully automated versions of the HHpred server that participated in the community-wide blind protein structure prediction competition CASP8. What makes HHpred unique is the combination of usability, short response times (typically under 15 min) and a model accuracy that is competitive with those of the best servers in CASP8.


The relationship of community activities with cognitive impairment and depressive mood independent of mobility disorder in Japanese older adults.
This study aimed to examine the relationship of participating in community activities (CA) with cognitive impairment and depressive mood independent of mobility disorder (MD) among older Japanese people. Elderly residents in institutions or those requiring long-term care insurance services were excluded; questionnaires were mailed to 5401 older adults in 2013. The response rate was 94.3% (n=5094). We used multiple imputation to manage missing data. The questionnaire addressed physical fitness, memory, mood, and CA. Participants were divided into two groups (good and bad) based on the median scores for physical fitness, memory, and mood. We identified items related to periodically performed CA, cognitive impairment, and depressive mood, and examined correlations between scores on these sets of items. The mean age was 75.9 years; 58.4% of participants were women. The following CA significantly predicted reduced cognitive impairment and depressive mood independent of MD: volunteer activity, community activity, visiting friends at home, pursuing hobbies, paid work, farm work, and daily shopping. These results were corrected for age, sex, and response method (mail or home-visit). Higher CA scores were associated with lower cognitive impairment and lower depressive mood independent of MD. CA is negatively associated with cognitive impairment and depressive mood among community-dwelling elderly independent of MD; promoting CA may protect against cognitive impairment and depressive mood in this population. However, MD, cognitive impairment, and depressive mood may lead to reduced CA.


Antiobesity Effect of Exopolysaccharides Isolated from Kefir Grains.
Physiological properties of water-soluble exopolysaccharides (EPS) and residues after EPS removal (Res) from the probiotic kefir were determined in high-fat (HF) diet-fed C57BL/6J mice. EPS solutions showed rheological properties and lower viscosity compared to those of β-glucan (BG). EPS significantly suppressed the adipogenesis of 3T3-L1 preadipocytes in a dose-dependent manner. Mice were fed HF diets containing 5% EPS, 5% BG, 8% Res, or 5% microcrystalline cellulose (control) for 4 weeks. Compared with the control, EPS supplementation significantly reduced HF diet-induced body weight gain, adipose tissue weight, and plasma very-low-density lipoprotein cholesterol concentration (P < 0.05). Res and BG significantly reduced body weight gain; however, reduction in adipose tissue weight was not statistically significant, suggesting that the antiobesity effect of EPS occurs due to viscosity and an additional factor. EPS supplementation significantly enhanced abundance of Akkermansia spp. in feces. These data indicate that EPS shows significant antiobesity effects possibly via intestinal microbiota alterations.


Mechanisms of estrogen receptor signaling: convergence of genomic and nongenomic actions on target genes.
Estrogen receptors (ERs) act by regulating transcriptional processes. The classical mechanism of ER action involves estrogen binding to receptors in the nucleus, after which the receptors dimerize and bind to specific response elements known as estrogen response elements (EREs) located in the promoters of target genes. However, ERs can also regulate gene expression without directly binding to DNA. This occurs through protein-protein interactions with other DNA-binding transcription factors in the nucleus. In addition, membrane-associated ERs mediate nongenomic actions of estrogens, which can lead both to altered functions of proteins in the cytoplasm and to regulation of gene expression. The latter two mechanisms of ER action enable a broader range of genes to be regulated than the range that can be regulated by the classical mechanism of ER action alone. This review surveys our knowledge about the molecular mechanism by which ERs regulate the expression of genes that do not contain EREs, and it gives examples of the ways in which the genomic and nongenomic actions of ERs on target genes converge. Genomic and nongenomic actions of ERs that do not depend on EREs influence the physiology of many target tissues, and thus, increasing our understanding of the molecular mechanisms behind these actions is highly relevant for the development of novel drugs that target specific receptor actions.


A Tale of Two Temporal Coding Strategies: Common and Dissociable Brain Regions Involved in Recency versus Associative Temporal Order Retrieval Strategies.
Numerous studies indicate the importance of the hippocampus to temporal order retrieval. However, behavioral studies suggest that there are different ways to retrieve temporal order information from encoded sequences, one involving an associative strategy (retrieving associations using neighboring items in a list) and another involving a recency strategy (determining which of two items came first). It remains unresolved, however, whether both strategies recruit the hippocampus or only associative strategies, consistent with the hippocampus's role in relational processing. To address this, we developed a paradigm in which we dissociated associative versus recency-based retrieval, involving the same stimulus presentation during retrieval. Associative retrieval involved an increase in RT (and decrease in performance) with greater distances between intervals, consistent with the need to retrieve intervening associations. Recency-based retrieval involved an increase in RT (and decrease in performance) with shorter distances between intervals, suggesting the use of a strength-based coding mechanism to retrieve information. We employed fMRI to determine the neural basis of the different strategies. Both strategies showed significant levels of hippocampal activation and connectivity that did not differ between tasks. In contrast, both univariate and connectivity pattern analyses revealed differences in extrahippocampal areas such as parietal and frontal cortices. A covariate analysis suggested that differences could not be explained by task difficulty alone. Together, these findings suggest that the hippocampus plays a role in both forms of temporal order retrieval, with neocortical networks mediating the different cognitive demands for associative versus recency-based temporal order retrieval.


Low-income women with abnormal breast findings: results of a randomized trial to increase rates of diagnostic resolution.
Timely diagnostic resolution of abnormal breast findings represents a critical step in efforts to reduce breast cancer morbidity and mortality. Yet, follow-up rates among resource poor populations are not optimal. Efforts to mitigate this disparity are needed. We report results of a randomized trial assessing the effectiveness of a patient support and navigation intervention in increasing timely diagnostic resolution of abnormal breast findings among indigent women. Women (n = 1,708) diagnosed with a breast abnormality at two public hospitals were randomized to an intervention or control group. The intervention, delivered through telephone, involved one call from a professional health worker and multiple calls from a lay health worker. The outcome, timely diagnostic resolution, defined as receipt of a definitive diagnosis (malignant or benign) within 6 months of the index referral, was assessed through medical chart audit. Intent-to-treat analyses revealed no significant effect of the intervention on timely diagnostic resolution. Diagnostic resolution rates were 55% and 56%, respectively, in the intervention and control arms. The significant predictors were method of abnormality identification (odds ratio = 1.50) and location of first scheduled appointment (odds ratio = 0.62). The intervention was not effective in creating change within the County health system. Achieving optimum diagnostic follow-up may require more intensive interventions than the one tested. In addition, system-level rather than patient-level interventions may hold more promise. There are no randomized trials reported in the literature testing interventions to increase diagnostic follow-up of breast abnormalities. Future research might test patient and system-level interventions that can be sustained beyond the study period.


Simultaneous orbital decompression and correction of upper eyelid retraction versus staged procedures in thyroid-related orbitopathy.
To evaluate the outcome of eyelid retraction surgery in thyroid-related orbitopathy (TRO) patients in 2 different surgical settings: done simultaneously with orbital decompression or as a staged procedure after orbital decompression. Retrospective, comparative, nonrandomized clinical study. Ninety-six patients (158 eyes). A review of electronic medical records of TRO patients who underwent surgery for upper eyelid retraction and orbital decompression at the Jules Stein Eye Institute in 1999 to 2003 was performed. Data regarding eyelid position, comprehensive eye examination, surgical outcome, and complications were analyzed. Anatomical and functional success based on margin reflex distance (MRD1; < or = 5 mm was graded as mild retraction; > 5 mm and < 7 mm, moderate; and > 7 mm, severe), and patients' discomfort. One hundred fifty-eight eyelid retraction surgeries were performed on 96 TRO patients (18 male and 78 female; mean age, 48 years); mean follow up time was 15 (+/-12) months. Group 1 consisted of patients undergoing simultaneous eyelid retraction surgery and orbital decompression and comprised 97 cases (surgeries). Group 2 included 61 cases of staged surgery: orbital decompression and eyelid retraction at a later stage. The groups had similar surgical outcomes, and > 85% had a better eyelid position postoperatively. Reoperation rates for residual or recurrent eyelid retraction were similar, overcorrection was higher in group 2 (5% vs. 0%, P = 0.03). Changes in MRD1, lagophthalmos, and exophthalmos were similar (P > 0.05, independent samples t test). Correction of eyelid retraction was effective in treating patients' discomfort and exposure keratopathy (P = 0.04, chi2). No severe complications occurred after orbital decompression or eyelid retraction surgery in this group of patients. Transconjunctival Muller's muscle recession for correction of eyelid retraction in mild to moderate TRO patients, performed simultaneously with deep lateral wall orbital decompression, resulted in acceptable eyelid position in two thirds of our patients. Overcorrection and consecutive ptosis occurred less often after combined orbital decompression and eyelid retraction surgery than after isolated eyelid repositioning surgery. If confirmed in prospective controlled studies, eyelid-repositioning surgery performed at the time of orbital decompression may decrease the number of total procedures and compress the time needed for surgical rehabilitation.


Racial/ethnic differences in circulating natriuretic peptide levels: The Diabetes Prevention Program.
Natriuretic peptides are cardiac-derived hormones that enhance insulin sensitivity and reduce fat accumulation. Low natriuretic peptide levels are associated with increased risk of type 2 diabetes mellitus (DM2); a condition with variable prevalence across racial/ethnic groups. Few studies have examined whether circulating natriuretic peptide levels and their response to preventive interventions for DM2 differ by race/ethnicity. The Diabetes Prevention Program (DPP) is a clinical trial (July 31, 1996- July 31, 2001) that randomized participants to preventive interventions for DM2. Using stored serum samples, we examined N-terminus pro-B-type natriuretic peptide (NT-proBNP) levels in 3,220 individuals (56% white; 19% African-American; 15% Hispanic; 5% American-Indian; 5% Asian). The influence of race/ethnicity on NT-proBNP concentrations at baseline and after two years of treatment with placebo, lifestyle, or metformin was examined with multivariable-adjusted regression. At baseline, NT-proBNP differed significantly by race (P < .001), with the lowest values in African-American individuals. Hispanic individuals also had lower baseline NT-proBNP levels compared with whites (P< .001), while NT-proBNP levels were similar between white, American-Indian, and Asian individuals. At two years of follow-up, NT-proBNP levels decreased in African-Americans in each of the DPP study arms, whereas they were stable or increased in the other racial/ethnic groups. In the DPP, African-American individuals had lower circulating NT-proBNP levels compared with individuals in other racial/ethnic groups at baseline and after two years of preventive interventions. Further studies should examine the cardio-metabolic implications of lower natriuretic peptide levels in African-Americans. Trial Registration: ClinicalTrials.gov NCT00004992.


Honey Bee and Bumble Bee Antiviral Defense.
Bees are important plant pollinators in both natural and agricultural ecosystems. Managed and wild bees have experienced high average annual colony losses, population declines, and local extinctions in many geographic regions. Multiple factors, including virus infections, impact bee health and longevity. The majority of bee-infecting viruses are positive-sense single-stranded RNA viruses. Bee-infecting viruses often cause asymptomatic infections but may also cause paralysis, deformity or death. The severity of infection is governed by bee host immune responses and influenced by additional biotic and abiotic factors. Herein, we highlight studies that have contributed to the current understanding of antiviral defense in bees, including the Western honey bee (Apis mellifera), the Eastern honey bee (Apis cerana) and bumble bee species (Bombus spp.). Bee antiviral defense mechanisms include RNA interference (RNAi), endocytosis, melanization, encapsulation, autophagy and conserved immune pathways including Jak/STAT (Janus kinase/signal transducer and activator of transcription), JNK (c-Jun N-terminal kinase), MAPK (mitogen-activated protein kinases) and the NF-κB mediated Toll and Imd (immune deficiency) pathways. Studies in Dipteran insects, including the model organism Drosophila melanogaster and pathogen-transmitting mosquitos, provide the framework for understanding bee antiviral defense. However, there are notable differences such as the more prominent role of a non-sequence specific, dsRNA-triggered, virus limiting response in honey bees and bumble bees. This virus-limiting response in bees is akin to pathways in a range of organisms including other invertebrates (i.e., oysters, shrimp and sand flies), as well as the mammalian interferon response. Current and future research aimed at elucidating bee antiviral defense mechanisms may lead to development of strategies that mitigate bee losses, while expanding our understanding of insect antiviral defense and the potential evolutionary relationship between sociality and immune function.


Single-molecule three-color FRET.
Fluorescence resonance energy transfer (FRET) measured at the single-molecule level can reveal conformational changes of biomolecules and intermolecular interactions in physiologically relevant conditions. Thus far single-molecule FRET has been measured only between two fluorophores. However, for many complex systems, the ability to observe changes in more than one distance is desired and FRET measured between three spectrally distinct fluorophores can provide a more complete picture. We have extended the single-molecule FRET technique to three colors, using the DNA four-way (Holliday) junction as a model system that undergoes two-state conformational fluctuations. By labeling three arms of the junction with Cy3 (donor), Cy5 (acceptor 1), and Cy5.5 (acceptor 2), distance changes between the donor and acceptor 1, and between the donor and acceptor 2, can be measured simultaneously. Thus we are able to show that the acceptor 1 arm moves away from the donor arm at the same time as the acceptor 2 arm approaches the donor arm, and vice versa, marking the first example of observing correlated movements of two different segments of a single molecule. Our data further suggest that Holliday junction does not spend measurable time with any of the helices unstacked, and that the parallel conformations are not populated to a detectable degree.


Neural hyperactivity related to working memory in drug-naive boys with attention deficit hyperactivity disorder.
Impaired working memory is thought to be a core feature of attention deficit hyperactivity disorder (ADHD). Previous imaging studies investigating working memory in ADHD have used tasks involving different cognitive resources and ignoring the categorical judgments about objects that are essential parts of performance in visual working memory tasks, thus complicating the interpretation of their findings. In the present study, we explored differential neural activation in children and adolescents with ADHD and in healthy controls using functional magnetic resonance imaging (fMRI) with the categorical n-back task (CN-BT), which maximized demands for executive reasoning while holding memory demands constant. A total of 33 drug-naive, right-handed male ADHD without comorbidity (mean age 9.9±2.4 years) and 27 right-handed, healthy male controls (mean age 10.9±2.7 years) were recruited in the present study. Event-related fMRI was used to study differences in brain activity during the CN-BT between the two groups. The two groups did not differ in their accuracy in the CN-BT, although the ADHD patients showed significantly shorter reaction times to correct responses than did the controls. During the CN-BT, both ADHD patients and controls showed significant positive and negative activations by the correct responses, mainly in the sensory-motor pathways and the striato-cerebellum circuit. Additionally, the ADHD patients showed significantly higher activation in the bilateral globus pallidus and the right hippocampus compared with the controls. There was also a positive correlation between hyperactivation of the left globus pallidus and the reaction time to correct responses in ADHD. In contrast to controls, ADHD patients showed neural hyperactivation in the striatum and mediotemporal areas during a working memory task involving categorization. Hyperfunction in these areas might be the pathophysiological foundation of ADHD, related to the deficits of working memory and the impulsive symptoms.


Temporal Trends in Polypharmacy and Hyperpolypharmacy in Older New Zealanders over a 9-Year Period: 2005–2013.
Polypharmacy and hyperpolypharmacy are proxy indicators for inappropriate medicine use. Inappropriate medicine use in older people leads to adverse clinical outcomes. The objectives of this study were to investigate the prevalence and trends of polypharmacy and hyperpolypharmacy in older people in New Zealand from 2005 to 2013, analyzing the pharmaceutical collections maintained by the Ministry of Health. A repeated cross-sectional analysis of population-level dispensing data was conducted from January 1, 2005 to December 31, 2013. Polypharmacy and hyperpolypharmacy in individuals were defined as the use of 5-9 medicines and ≥10 medicines, respectively, dispensed concurrently for a period of ≥90 days. Differences in polypharmacy and hyperpolypharmacy between 2005 and 2013 were examined. A multinomial regression model was used to predict sociodemographic characteristics associated with polypharmacy and hyperpolypharmacy. Polypharmacy and hyperpolypharmacy were found to be higher in 2013 compared to 2005 (polypharmacy: 29.5 vs. 23.4%, p<0.001; hyperpolypharmacy: 2.1 vs. 1.3%, p<0.001). The risk of polypharmacy and hyperpolypharmacy was higher in females, in those aged 80-84 years, in the Māori population (for polypharmacy) and the Middle Eastern, Latin American, or African population (for hyperpolypharmacy), in people living in the Southern-district health board, and in individuals with increasing deprivation. The population of New Zealand is aging and the number of older people with multiple chronic conditions is increasing. The proportion of older people exposed to polypharmacy and hyperpolypharmacy has increased in 2013 compared to 2005. Our study provides important information to alert health policy makers, researchers, and clinicians about the dire need to reduce the medication burden in older New Zealanders.


Nonlinear phase matching locking via optical readout.
For optimal Chi((2)) nonlinear interaction the phase matching condition must be satisfied. For type I and type II phase matched materials, this is generally achieved by controlling the temperature of the nonlinear media. We describe a technique to readout the phase-matching condition interferometrically, and experimentally demonstrate feedback control in a degenerate optical parametric amplifier (OPA) which is resonant at both the fundamental and harmonic frequencies. The interferometric readout technique is based on using the cavity resonances at the fundamental and harmonic frequencies to enable the readout of the phase mismatch. We achieve relatively fast temperature feedback using the photothermal effect, by modulating the amplitude of the OPA pump beam.


Effect of resin adhesive systems on root caries formation in vitro.
To evaluate the capability of different adhesive systems to inhibit in vitro caries formation. Four materials were tested: Gluma Comfort Bond (GL; Heraeus Kulzer), Gluma Comfort Bond + Desensitizer (GL+; Heraeus Kulzer), iBond (iB; Heraeus Kulzer), and One-up Bond F (OUB; Tokuyama). Bovine roots were ground to obtain flat mesial and distal dentin surfaces. Nail varnish was applied to the entire root surface except for two 10 mm X 2 mm windows on the flattened surfaces. The adhesives were applied to the exposed areas according to the manufacturers' instructions. The roots were incubated at 37 degrees C for 1 week in a suspension of StreptococcuS mutans and Lactobacillus acidophilus. Control groups either received no root surface treatment or were etched with 37% phosphoric acid (without immersion in the bacterial suspension). Specimens were sectioned perpendicular to the long axis of the roots, stained for 24 hours with Rhodamine B, and analyzed with confocal laser microscopy. The data were analyzed by 1-way ANOVA and Fisher's PLSD test at 95% confidence level. Lesions in the OUB, iB, and GL+ groups were significantly shallower than those in the no-treatment group. Acid etching did not produce any measurable lesions. Mean lesion depth in the GL group was similar to that in the no-treatment group. There was no significant difference between mean lesion depths in the GL+, GL, and iB groups. Fluoride- and glutaraldehyde-containing adhesive systems might be an aid in root caries prevention.


Surveillance for potential adverse events associated with the use of intravitreal bevacizumab for retinal and choroidal vascular disease.
To systematically study potential adverse events associated with the use of intraocular bevacizumab at a single medical center. Retrospective study of all consecutive patients receiving intraocular bevacizumab injections at the Stanford University Department of Ophthalmology between November 15, 2005 and July 14, 2006. Bevacizumab was given for exudative age-related macular degeneration, retinal vascular occlusion, diabetic macular edema, neovascular glaucoma, and five other indications. We analyzed medical records of 186 subjects (203 eyes) who received a total of 578 injections of 1.25 mg of bevacizumab. The average follow-up was approximately 6 months. Five eyes with exudative age-related macular degeneration developed retinal pigment epithelial (RPE) tears, all with preexisting RPE detachments. These five eyes represented 2.9% of all age-related macular degeneration eyes treated and 7% of the age-related macular degeneration eyes with preexisting RPE detachments at initiation of treatment. Other adverse events were rare and included retinal ischemia, subretinal hemorrhage, vitreous hemorrhage, ocular irritation or pain, worsened hypertension, and headache. No death or thromboembolic events were observed. Intraocular bevacizumab appears to be well tolerated for the treatment of a variety of retinal and choroidal vascular diseases. RPE tears may occur when treating choroidal neovascularization, particularly in patients with preexisting RPE detachment.


Plant Growth Environments with Programmable Relative Humidity and Homogeneous Nutrient Availability.
We describe the design, characterization, and use of "programmable", sterile growth environments for individual (or small sets of) plants. The specific relative humidities and nutrient availability experienced by the plant is established (RH between 15% and 95%; nutrient concentration as desired) during the setup of the growth environment, which takes about 5 minutes and <1$ in disposable cost. These systems maintain these environmental parameters constant for at least 14 days with minimal intervention (one minute every two days). The design is composed entirely of off-the-shelf components (e.g., LEGO® bricks) and is characterized by (i) a separation of root and shoot environment (which is physiologically relevant and facilitates imposing specific conditions on the root system, e.g., darkness), (ii) the development of the root system on a flat surface, where the root enjoys constant contact with nutrient solution and air, (iii) a compatibility with root phenotyping. We demonstrate phenotyping by characterizing root systems of Brassica rapa plants growing in different relative humidities (55%, 75%, and 95%). While most phenotypes were found to be sensitive to these environmental changes, a phenotype tightly associated with root system topology-the size distribution of the areas encircled by roots-appeared to be remarkably and counterintuitively insensitive to humidity changes. These setups combine many of the advantages of hydroponics conditions (e.g., root phenotyping, complete control over nutrient composition, scalability) and soil conditions (e.g., aeration of roots, shading of roots), while being comparable in cost and setup time to Magenta® boxes.


Words matter: the prevalence of chiropractic-specific terminology on Australian chiropractors' websites.
Chiropractors use words and phrases in unique ways to express traditional, chiropractic-specific theories. This lexicon represents concepts that reinforce the separation of chiropractic from other health care professions. It may impact referrals both to and from chiropractors, lead to public confusion about health care issues, and reduce cross-disciplinary research. Therefore, it is important to understand how prevalent chiropractic-specific terms are in publicly available media. Five chiropractic terms were selected: subluxation, adjustment, vital (-ism/-istic), wellness, and Innate (Intelligence). States and territories in Australia were proportionately sampled according to population of chiropractors using a Google search for chiropractors' private practice websites. The top results were recorded. Websites were word-searched on every publicly available page for the five terms. Context was checked to count only terms that were used to support a chiropractic-specific concepts. The number of occurrences of each term was recorded, tallied nationally and by state/territory. Descriptive statistics were applied to determine prevalence. Three hundred sixty-nine websites were sampled, based on an estimate of 5500 chiropractors practising in Australia. Nationally, 85% of chiropractors used one or more terms. The term adjust (-ing/-ment) occurred most frequently, being found on 283 websites (77%) with a total of 2249 occurrences. Wellness was found on 199 websites (54%) with 872 occurrences; subluxation was found on 104 websites (28%), 489 occurrences; vital (-ism/-istic) on 71 websites (19%) with 158 occurrences; and Innate was least used, being found on 39 websites (11%) with 137 occurrences. A majority of the Australian chiropractors sampled used one or more chiropractic-specific terms on their websites. Future research should explore the effects of chiropractic language on the public, policy-makers, and other health care professionals.


Low-dose studies of bystander cell killing with targeted soft X rays.
The Gray Cancer Institute ultrasoft X-ray microprobe was used to quantify the bystander response of individual V79 cells exposed to a focused carbon K-shell (278 eV) X-ray beam. The ultrasoft X-ray microprobe is designed to precisely assess the biological response of individual cells irradiated in vitro with a very fine beam of low-energy photons. Characteristic CK X rays are generated by a focused beam of 10 keV electrons striking a graphite target. Circular diffraction gratings (i.e. zone plates) are then employed to focus the X-ray beam into a spot with a radius of 0.25 microm at the sample position. Using this microbeam technology, the correlation between the irradiated cells and their nonirradiated neighbors can be examined critically. The survival response of V79 cells irradiated with a CK X-ray beam was measured in the 0-2-Gy dose range. The response when all cells were irradiated was compared to that obtained when only a single cell was exposed. The cell survival data exhibit a linear-quadratic response when all cells were targeted (with evidence for hypersensitivity at low doses). When only a single cell was targeted within the population, 10% cell killing was measured. In contrast to the binary bystander behavior reported by many other investigations, the effect detected was initially dependent on dose (<200 mGy) and then reached a plateau (>200 mGy). In the low-dose region (<200 mGy), the response after irradiation of a single cell was not significantly different from that when all cells were exposed to radiation. Damaged cells were distributed uniformly over the area of the dish scanned (approximately 25 mm2). However, critical analysis of the distance of the damaged, unirradiated cells from other damaged cells revealed the presence of clusters of damaged cells produced under bystander conditions.


S-shooting: a Bennett-Chandler-like method for the computation of rate constants from committor trajectories.
Mechanisms of rare transitions between long-lived stable states are often analyzed in terms of commitment probabilities, determined from swarms of short molecular dynamics trajectories. Here, we present a computer simulation method to determine rate constants from such short trajectories combined with free energy calculations. The method, akin to the Bennett-Chandler approach for the calculation of reaction rate constants, requires the definition of a valid reaction coordinate and can be applied to both under- and overdamped dynamics. We verify the correctness of the algorithm using a one-dimensional random walker in a double-well potential and demonstrate its applicability to complex transitions in condensed systems by calculating cavitation rates for water at negative pressures.


A population pharmacokinetic model is beneficial in quantifying hair concentrations of ritonavir-boosted atazanavir: a study of HIV-infected Zimbabwean adolescents.
Adolescents experience higher levels of non-adherence to HIV treatment. Drug concentration in hair promises to be reliable for assessing exposure to antiretroviral (ARV) drugs. Pharmacokinetic modelling can explore utility of drug in hair. We aimed at developing and validating a pharmacokinetic model based on atazanavir/ritonavir (ATV/r) in hair and identify factors associated with variabilities in hair accumulation. We based the study on secondary data analysis whereby data from a previous study on Zimbabwean adolescents which collected hair samples at enrolment and 3 months follow-up was used in model development. We performed model development in NONMEM (version 7.3) ADVAN 13. There is 16% / 18% of the respective ATV/r in hair as a ratio of steady-state trough plasma concentrations. At follow-up, we estimated an increase of 30% /42% of respective ATV/r in hair. We associated a unit increase in adherence score with 2% increase in hair concentration both ATV/r. Thinner participants had 54% higher while overweight had 21% lower atazanavir in hair compared to normal weight participants. Adolescents receiving care from fellow siblings had atazanavir in hair at least 54% less compared to other forms of care. The determinants of increased ATV/r concentrations in hair found in our analysis are monitoring at follow up event, body mass index, and caregiver status. Measuring drug concentration in hair is feasibly accomplished and could be more accurate for monitoring ARV drugs exposure.


Terahertz waveguiding between parallel dielectric films.
We report a study of terahertz waveguiding between parallel dielectric films, a system that can be viewed as a planar analogue of hollow core fibres that exploit anti-resonant reflection optical waveguiding (ARROW). With the aid of time domain waveguide mode imaging, the frequency dependent transition from ARROW to total internal reflection guiding in the individual films as the film separation is reduced and the effect on the transmission of adding variably spaced cladding layers are clearly revealed. Good agreement for the transmission, dispersion and loss is obtained with simple analytical models for film separations greater than about five wavelengths suggesting that the same models could be usefully used to predict the behavior in the case of the technologically more important cylindrical geometry.


[An active search for cases of zoonotic visceral leishmaniasis in indigenous Colombian children and the canine population].
Carrying out an active search for cases of canine and human zoonotic visceral leishmaniasis (LVZ) by determining IgG antibodies against Leishmania infantum by indirect immunofluorescence (IFAT) and assessing the risk factors associated with the disease occurring in a Colombian endemic area. 580 indigenous children aged less than five and 270 cross-bred dogs from 5 rural areas near the town of Coyaima (Tolima) were evaluated by determining their antibodies using the Colombian Leishmania infantum (infantum) MOHOM/ COL/CLO44B strain as antigen. 527 households in the area were surveyed to assess the risk factors and protective measures being taken. No sero reactivity to Leishmania infantum antigens by IFAT test was observed in the child population. Leishmania infantum antibodies were observed in 68.5 % of the dogs (185/270). The survey found deficiencies in the housing's sanitary conditions. The use of insecticide-impregnated bed-nets was evident in 48 % (130/270) of the dwellings, as were poor knowledge of the disease and a low demand for traditional medical services (41.5 %). Control measures must be strengthened for interrupting the reservoir-vector-human transmission chain, evaluating health authorities' strategies for promoting protective habits and improving living conditions and the susceptible population's environment.


A randomized trial of controlled-release oxycodone during inpatient rehabilitation following unilateral total knee arthroplasty.
Reliance on "as-needed" analgesia following total knee arthroplasty may lead to inadequate control of pain and delayed recovery of function. Preemptive use of controlled-release opioids may improve pain control, accelerate recovery, and reduce the need for inpatient rehabilitative services. This study was designed to determine whether controlled-release opioids enhance post-arthroplasty pain control and facilitate functional recovery during rehabilitation. Fifty-nine patients admitted for inpatient rehabilitation following unilateral total knee arthroplasty were randomized to receive OxyContin (controlled-release oxycodone) (twenty-nine patients) or a placebo (thirty patients) every twelve hours. Both groups could receive on-request, immediate-release oxycodone (5 mg every four hours). The dose of study medication was increased on the basis of the frequency of requests for immediate-release oxycodone. Measures of interest included pain ratings as determined with a visual-analog scale, changes in the range of motion of the knee and quadriceps strength, and improvements in selected Functional Independence Measure scores during the first eight physical therapy sessions. The duration of the hospital stay for rehabilitation also was compared between the two groups. Baseline demographic, clinical, and functional characteristics were similar between the OxyContin and placebo groups. Compared with the placebo group, the patients who received OxyContin reported significantly less pain as well as significantly greater range of motion of the knee (passive motion, p = 0.036; active motion, p< 0.001) and quadriceps strength (p = 0.001) by the eighth physical therapy session. The patients who received OxyContin also were discharged from the rehabilitation hospital at an average of 2.3 days earlier than the patients in the placebo group (p = 0.013). Preemptive use of controlled-release oxycodone during rehabilitation following total knee arthroplasty leads to improved pain control, more rapid functional recovery, and a reduced need for inpatient rehabilitative services.


A novel sequence to improve auditory functional MRI with variable silent delays.
Auditory functional MRI (fMRI) often uses silent inter-volume delays for stimulus presentation. However, maintaining the steady-state of the magnetization usually requires constant delays. Here, a novel acquisition scheme dubbed "pre-Saturated EPI using Multiple delays in Steady-state" (SEPIMS) is proposed, using spin saturation at a fixed delay before each volume to maintain steady-state conditions, independent of previous spin history. This concept allows for variable inter-volume delays and thus for flexible stimulus design in auditory fMRI. The purpose was to compare the signal stability of SEPIMS and conventional sparse EPI (CS-EPI). The saturation module comprises two non-selective adiabatic saturation pulses. The efficiency of the saturation and its effect on the SEPIMS signal stability is tested in vitro and in vivo. Data show that SEPIMS yields the same signal stability as CS-EPI, even for extreme variations between inter-volume delay durations. However, dual saturation pulses are required to achieve sufficiently high saturation efficiency in compartments with long T1 values. Importantly, spoiler gradient pulses after the EPI readout have to be optimized to avoid eddy-current-induced image distortions. The proposed SEPIMS sequence maintains high signal stability in the presence of variable inter-volume durations, thus allowing for flexible stimulus design.


Efficient multicolor tunability of ultrasmall ternary-doped LaF3 nanoparticles: energy conversion and magnetic behavior.
Luminescence-tunable multicolored LaF3:xCe3+,xGd3+,yEu3+ (x = 5; y = 1, 5, 10, and 15 mol%) nanoparticles have been synthesized via a low cost polyol method. Powder X-ray diffraction and high-resolution transmission electron microscopy studies confirm the hexagonal phase of the LaF3:xCe3+,xGd3+,yEu3+ nanophosphors with average sizes (oval shape) ranging from 5 to 7 nm. Energy-dispersive X-ray spectroscopy analyses show the uniform distribution of Ce3+, Gd3+, and Eu3+ dopants in the LaF3 host matrix. The photoluminescence spectra and electron paramagnetic resonance measurements guarantee the presence of Eu2+, corroborated through DC susceptibility measurements of the samples displaying paramagnetic behavior at 300 K, whereas weak ferromagnetic ordering is shown at 2 K. The non-radiative energy transfer processes from the 4f(2F5/2) → 5d state (Ce3+) to the intraconfigurational 4f excited levels of rare earth ions and simultaneous emissions in the visible region from the 4f65d1 (Eu2+) and 5D0 (Eu3+) emitting levels, leading to overlapped broad and narrow emission bands, have been proclaimed. The energy transfer mechanism proposes involvement of the Gd3+ ion sub-lattice as the bridge and finally trapping by Eu2+/3+, upon excitation of the Ce3+ ion. The calculation of experimental intensity parameters (Ω2,4) has been discussed and the highest emission quantum efficiency (η = 85%) of the Eu3+ ion for the y = 10 mol% sample is reported. The advantageous existence of the Eu2+/Eu3+ ratio along with variously doped nanomaterials described in this work, results in tunable emission color in the blue-white-red regions, highlighting the potential application of the samples in solid-state lighting devices, scintillation devices, and multiplex detection.


Inverse finite element characterization of the human myometrium derived from uniaxial compression experiments.
The low strain-rate behavior of the human myometrium under compression was determined. To this end, uniaxial, unconstrained compression experiments were conducted on a total of 25 samples from three excised human uteri at strain rates between 0.001 s(-1) and 0.008 s(-1). A three-dimensional finite element model of each sample was created and used together with an optimization algorithm to find material parameters in an inverse estimation process. Friction and shape irregularities of samples were incorporated in the models. The uterine specimens in compression were modeled as viscoelastic, non-linear, nearly incompressible and isotropic continua. Simulations of uniaxial, frictionless compressions of an idealized cuboid were used to compare the resulting material parameters among each other. The intra- and inter-subject variability in stiffness of specimens was found to be large and to cover such a wide range that the effect of anisotropy which is of minor influence under compressive deformations in the first place could be neglected. Material parameters for a viscoelastic model based on a decoupled, reduced quadratic strain-energy function were presented for the uterine samples representing a median stiffness.


Signatures of positive selection apparent in a small sample of human exomes.
Exome sequences, which comprise all protein-coding regions, are promising data sets for studies of natural selection because they offer unbiased genome-wide estimates of polymorphism while focusing on the portions of the genome that are most likely to be functionally important. We examine genomic patterns of polymorphism within 10 diploid autosomal exomes of European and African descent. Using coalescent simulations, we show how polymorphism, site frequency spectra, and intercontinental divergence in these samples would be influenced by different modes of positive selection. We examine putatively selected loci from four previous genome-wide scans of SNP genotypes and demonstrate that these regions indeed show unusual population genetic patterns in the exome data. Using a series of conservative criteria based on exome polymorphism, we are able to fine-scale map signatures of selection, in many cases pinpointing a single candidate SNP. We also identify and evaluate novel candidate selection genes that show unusual patterns of polymorphism. We sequence a portion of one novel candidate locus, IVL, in 74 individuals from multiple continents and examine global genetic diversity. Thus, we confirm, narrow, and supplement existing catalogs of putative targets of selection, and show that exome data sets, which are likely to soon become common, will be powerful tools for identifying adaptive genetic variation.


Cross-talks via mTORC2 can explain enhanced activation in response to insulin in diabetic patients.
The molecular mechanisms of insulin resistance in Type 2 diabetes have been extensively studied in primary human adipocytes, and mathematical modelling has clarified the central role of attenuation of mammalian target of rapamycin (mTOR) complex 1 (mTORC1) activity in the diabetic state. Attenuation of mTORC1 in diabetes quells insulin-signalling network-wide, except for the mTOR in complex 2 (mTORC2)-catalysed phosphorylation of protein kinase B (PKB) at Ser473 (PKB-S473P), which is increased. This unique increase could potentially be explained by feedback and interbranch cross-talk signals. To examine if such mechanisms operate in adipocytes, we herein analysed data from an unbiased phosphoproteomic screen in 3T3-L1 adipocytes. Using a mathematical modelling approach, we showed that a negative signal from mTORC1-p70 S6 kinase (S6K) to rictor-mTORC2 in combination with a positive signal from PKB to SIN1-mTORC2 are compatible with the experimental data. This combined cross-branch signalling predicted an increased PKB-S473P in response to attenuation of mTORC1 - a distinguishing feature of the insulin resistant state in human adipocytes. This aspect of insulin signalling was then verified for our comprehensive model of insulin signalling in human adipocytes. Introduction of the cross-branch signals was compatible with all data for insulin signalling in human adipocytes, and the resulting model can explain all data network-wide, including the increased PKB-S473P in the diabetic state. Our approach was to first identify potential mechanisms in data from a phosphoproteomic screen in a cell line, and then verify such mechanisms in primary human cells, which demonstrates how an unbiased approach can support a direct knowledge-based study.


Marine reserves lag behind wilderness in the conservation of key functional roles.
Although marine reserves represent one of the most effective management responses to human impacts, their capacity to sustain the same diversity of species, functional roles and biomass of reef fishes as wilderness areas remains questionable, in particular in regions with deep and long-lasting human footprints. Here we show that fish functional diversity and biomass of top predators are significantly higher on coral reefs located at more than 20 h travel time from the main market compared with even the oldest (38 years old), largest (17,500 ha) and most restrictive (no entry) marine reserve in New Caledonia (South-Western Pacific). We further demonstrate that wilderness areas support unique ecological values with no equivalency as one gets closer to humans, even in large and well-managed marine reserves. Wilderness areas may therefore serve as benchmarks for management effectiveness and act as the last refuges for the most vulnerable functional roles.


Oto-facio-cervical (OFC) syndrome is a contiguous gene deletion syndrome involving EYA1: molecular analysis confirms allelism with BOR syndrome and further narrows the Duane syndrome critical region to 1 cM.
Branchio-oto-renal (BOR) syndrome is an autosomal dominant disorder involving hearing loss, branchial defects, ear pits and renal abnormalities. Oto-facio-cervical (OFC) syndrome is clinically similar to BOR syndrome, with clinical features in addition to those of BOR syndrome. Mutations in the EYA1 gene (localised to 8q13.3) account for nearly 70% of BOR syndrome cases exhibiting at least three of the major features. Small intragenic deletions of the 3' region of the gene have also been reported in patients with BOR syndrome. We have developed a fluorescent quantitative multiplex polymerase chain reaction for three 3' exons (7, 9 and 13) of the EYA1 gene. This dosage assay, combined with microsatellite marker analysis, has identified de novo deletions of the EYA1 gene and surrounding region in two patients with complex phenotypes involving features of BOR syndrome. One patient with OFC syndrome carried a large deletion of the EYA1 gene region, confirming that OFC syndrome is allelic with BOR syndrome. Microsatellite analysis has shown that comparison of the boundaries of this large deletion with other reported rearrangements of the region reduces the critical region for Duane syndrome (an eye movement disorder) to between markers D8S553 and D8S1797, a genetic distance of approximately 1 cM.


Mapping and quantifying mammalian transcriptomes by RNA-Seq.
We have mapped and quantified mouse transcriptomes by deeply sequencing them and recording how frequently each gene is represented in the sequence sample (RNA-Seq). This provides a digital measure of the presence and prevalence of transcripts from known and previously unknown genes. We report reference measurements composed of 41-52 million mapped 25-base-pair reads for poly(A)-selected RNA from adult mouse brain, liver and skeletal muscle tissues. We used RNA standards to quantify transcript prevalence and to test the linear range of transcript detection, which spanned five orders of magnitude. Although >90% of uniquely mapped reads fell within known exons, the remaining data suggest new and revised gene models, including changed or additional promoters, exons and 3' untranscribed regions, as well as new candidate microRNA precursors. RNA splice events, which are not readily measured by standard gene expression microarray or serial analysis of gene expression methods, were detected directly by mapping splice-crossing sequence reads. We observed 1.45 x 10(5) distinct splices, and alternative splices were prominent, with 3,500 different genes expressing one or more alternate internal splices.


Spondyloepiphyseal dysplasia tarda with progressive arthropathy.
Spondyloepiphyseal dysplasia tarda with progressive arthropathy (SEDT-PA) is a rare autosomal recessive skeletal dysplasia affecting primarily the articular cartilage. Here we present a nine-year-old girl from Middle Anatolia (Sivas) with SEDT-PA. Her complaints were pain and progressive deformity of the joints. She had a short stature with increased thoracic kyphosis and lumbar lordosis. The range of motion was limited in the spine and the peripheral joints and she had deformities. Radiologic examination revealed generalized platyspondyly and irregularity of the vertebral endplates. There was minimal joint space narrowing at proximal interphalangeal joints, but there were no bone erosions. Metaphyses were widened and epiphyses were squared in other joints with generalized osteopenia and severe osteoarthritic changes prominent in hips. Laboratory examination revealed a mild increase in acute phase reactants. Genetic disorders like SEDT-PA may also have rheumatological involvement, so they should be kept in mind in differential diagnosis of inflammatory joint diseases.


The value of an immunoenzymatic test (enzyme-linked immunosorbent assay) for the diagnosis of strongyloidiasis in patients immunosuppressed by hematologic malignancies.
The diagnosis of strongyloidiasis relies upon the identification of the parasite in stool samples. In 1981, a serologic assay was developed, which was useful in the diagnosis of strongyloidiasis in the immunocompetent host. In the present study, we evaluated the enzyme-linked immunosorbent assay (ELISA) in patients with hematologic malignancies. Between April 1995 and December 1998, sera from 164 consecutive patients were tested for the presence of IgG antibody to Strongyloides stercoralis. Patient was considered uninfected after at least three negative stool examinations. The prevalence of strongyloidiasis was 13%. The underlying diseases were acute leukemia in 21% and lymphoma in 52% of the patients. The majority of the patients were receiving chemotherapy (93%) and steroids (76%). The sensitivity, specificity, and positive and negative predictive values were 68%, 89%, 48%, and 95%, respectively. The ELISA may be an excellent assay to rule out the diagnosis of strongyloidiasis in patients with hematologic malignancies.


Optimization of the fermentation conditions of Rhizopus japonicus M193 for the production of chitin deacetylase and chitosan.
To improve the production of chitin deacetylase (CDA) for the bioconversion of chitin to chitosan with desirable functionality, the effect of the nutritional requirement on the CDA production from Rhizopus japonicus M193 fermentation was investigated under submerged conditions. Nutritional elements including glucose (g/L), inoculum level (%), and MgSO4·7H2O (g/L), as well as culture time (d) were identified as the most critical factors for the CDA production based on the results from Plackett-Burman design (PBD). Taguchi design with orthogonal array was further employed to optimize R. japonicus M193 fermentation conditions based on the results from PBD, in which 2.5% chitin, 5 g/L glucose, 5% inoculum level, 0.6g/L MgSO4·7H2O, and 5d culture time were identified as the optimal fermentation conditions. Under this condition, the maximum CDA production, DDA and MM of produced chitosan were 547.38 ± 12.06 U/L, 78.85 ± 1.68%, and 125.63 ± 3.74 kDa, respectively. Obtained chitosan displayed similar physicochemical and structural properties to those of commercial chitosan extracted using chemical method based on the results from Fourier transform infrared spectrometer (FT-IR), thermogravimetric analysis (TGA)-differential scanning calorimetry (DSC), and nuclear magnetic resonance (NMR) assays, while the use of chemical reagents was significantly reduced.


Systematic review of foodborne burden of disease studies: quality assessment of data and methodology.
Burden of disease (BoD) studies aim to identify the public health impact of different health problems and risk factors. To assess BoD, detailed knowledge is needed on epidemiology, disability and mortality in the population under study. This is particularly challenging for foodborne disease, because of the multitude of causative agents and their health effects. The purpose of this study is to systematically review the methodology of foodborne BoD studies. Three key questions were addressed: 1) which data sources and approaches were used to assess mortality, morbidity and disability?, 2) which methodological choices were made to calculate Disability Adjusted Life Years (DALY), and 3) were uncertainty analyses performed and if so, how? Studies (1990-June 2012) in international peer-reviewed journals and grey literature were identified with main inclusion criteria being that the study assessed disability adjusted life years related to foodborne disease. Twenty-four studies met our inclusion criteria. To assess incidence or prevalence of foodborne disease in the population, four approaches could be distinguished, each using a different data source as a starting point, namely 1) laboratory-confirmed cases, 2) cohort or cross-sectional data, 3) syndrome surveillance data and 4) exposure data. Considerable variation existed in BoD methodology (e.g. disability weights, discounting, age-weighting). Almost all studies analyzed the effect of uncertainty as a result of possible imprecision in the parameter values. Awareness of epidemiological and methodological rigor between foodborne BoD studies using the DALY approach is a critical priority for advancing burden of disease studies. Harmonization of methodology that is used and of modeling techniques and high quality data can enlarge the detection of real variation in DALY outcomes between pathogens, between populations or over time. This harmonization can be achieved by identifying substantial data gaps and uncertainty and establish which sequelae of foodborne disease agents should be included in BoD calculations.


Areca catechu-From farm to food and biomedical applications.
The family Arecaceae includes 181 genera and 2,600 species with a high diversity in physical characteristics. Areca plants, commonly palms, which are able to grow in nearly every type of habitat, prefer tropical and subtropical climates. The most studied species Areca catechu L. contains phytochemicals as phenolics and alkaloids with biological properties. The phenolics are mainly distributed in roots followed by fresh unripe fruits, leaves, spikes, and veins, while the contents of alkaloids are in the order of roots, fresh unripe fruits, spikes, leaves, and veins. This species has been reputed to provide health effects on the cardiovascular, respiratory, nervous, metabolic, gastrointestinal, and reproductive systems. However, in many developing countries, quid from this species has been associated with side effects, which include the destruction of the teeth, impairment of oral hygiene, bronchial asthma, or oral cancer. Despite these side effects, which are also mentioned in this work, the present review collects the main results of biological properties of the phytochemicals in A. catechu. This study emphasizes the in vitro and in vivo antioxidant, antimicrobial, anticancer, and clinical effectiveness in humans. In this sense, A. catechu have demonstrated effectiveness in several reports through in vitro and in vivo experiments on disorders such as antimicrobial, antioxidant, or anticancer. Moreover, our findings demonstrate that this species presents clinical effectiveness on neurological disorders. Hence, A. catechu extracts could be used as a bioactive ingredient for functional food, nutraceuticals, or cosmeceuticals. However, further studies, especially extensive and comprehensive clinical trials, are recommended for the use of Areca in the treatment of diseases.


Resolving the complexity of the human genome using single-molecule sequencing.
The human genome is arguably the most complete mammalian reference assembly, yet more than 160 euchromatic gaps remain and aspects of its structural variation remain poorly understood ten years after its completion. To identify missing sequence and genetic variation, here we sequence and analyse a haploid human genome (CHM1) using single-molecule, real-time DNA sequencing. We close or extend 55% of the remaining interstitial gaps in the human GRCh37 reference genome--78% of which carried long runs of degenerate short tandem repeats, often several kilobases in length, embedded within (G+C)-rich genomic regions. We resolve the complete sequence of 26,079 euchromatic structural variants at the base-pair level, including inversions, complex insertions and long tracts of tandem repeats. Most have not been previously reported, with the greatest increases in sensitivity occurring for events less than 5 kilobases in size. Compared to the human reference, we find a significant insertional bias (3:1) in regions corresponding to complex insertions and long short tandem repeats. Our results suggest a greater complexity of the human genome in the form of variation of longer and more complex repetitive DNA that can now be largely resolved with the application of this longer-read sequencing technology.


Male genital hygiene beliefs and practices in Nairobi, Kenya.
Attitudes and practices concerning genital hygiene may influence topical microbicide use by men. This study examines knowledge and behaviours related to hygiene, genital hygiene, circumcision and hygiene, and to genital hygiene before and after sex among men in Nairobi, Kenya. We conducted 463 interviews of men recruited by respondent driven sampling techniques and 10 focus group discussions with a subsample of 100 volunteers from this group. Men were asked individual quantitative survey and qualitative group discussion questions about general hygiene behaviours, genital hygiene, and the temporal relation of genital hygiene behaviours to sexual encounters. Bathing once daily was associated with education, income, and inside tap water. Genital washing aside from regular bathing and washing before sex ever were negatively associated with bathroom crowding. Genital hygiene before the most recent sexual encounter was uncommon and negatively associated with HIV risk perception, bathroom crowding, and ethnicity. Men believed genital hygiene before sex would arouse suspicions of infidelity or cool sexual ardour. Genital hygiene after sex was associated with education, religion, and having inside tap water. Genital hygiene after the most recent sexual encounter was associated with age, income, and with men having at least one child. Genital hygiene behaviours were associated with resource access factors and group discussions suggest that they are modulated by interactions in sexual partnerships. Topical microbicides may improve hygiene before and after sex.


Phytophthora × pelgrandis Causes Root and Collar Rot of Lavandula stoechas in Italy.
In 2007, Phytophthora isolates with atypical morphological and biological characteristics were found associated with root and collar rot of potted plants of Stoechas lavender (Lavandula stoechas) in an ornamental nursery in Italy. A polyphasic approach, including morphological and cultural observations, sequencing the ITS-rDNA region, the Pheca and the mitochondrial coxI genes, multiplex PCRs with primers specific for P. nicotianae or P. cactorum, as well as random amplified polymorphic DNA-polymerase chain reaction, was used to characterize these isolates. On the basis of morpho-cultural and molecular analyses, the isolates from Stoechas lavender were identified as Phytophthora × pelgrandis, a natural hybrid of P. nicotianae × P. cactorum previously reported in other European countries, the Americas, and Taiwan, as a pathogen of ornamentals and loquat plants. In pathogenicity tests using potted plants of Stoechas lavender, the P. × pelgrandis isolates, similarly to the parental species P. nicotianae, induced the symptoms observed on plants with natural infections and were reisolated only from artificially inoculated plants. Dispersal of P. × pelgrandis on this host could exacerbate the damage caused by Phytophthora root and collar rot, of which the main causal agent presently is P. nicotianae on lavender in Europe. Application of hygienic measures is important to reduce the proliferation and spread of the Phytophthora hybrids.


The evolution of sleep is inevitable in a periodic world.
There are two contrasting explanations of sleep: as a proximate, essential physiological function or as a behavioral, adaptive state of inactivity and these hypotheses remain widely debated. To investigate the adaptive significance of sleep, we develop an evolutionary argument formulated as a tractable partial differential equation model. We allow demographic parameters such as birth and mortality rates to vary through time in both safe and vulnerable sleeping environments. From this model we analytically calculate population growth rate (fitness) for sleeping and non-sleeping strategies. We find that, in a temporally heterogeneous environment, sleep behavior always achieves a higher fitness than non-sleeping behavior. As organisms do not exist in constant environments, we conclude that the evolution of sleep is inevitable. Further, we suggest that the two contrasting theories need not be mutually exclusive.


Comparison of serum creatinine-based estimating equations with gates protocol for predicting glomerular filtration rate in indian population.
In clinical practice, serum creatinine-based predicting equations and Gates protocol based on gamma camera imaging of kidneys after injection of Tc99m-diethylenetriaminepentaacetic acid (DTPA) are commonly used to assess glomerular filtration rate (GFR). Comparison of these methods, especially the chronic kidney disease-epidemiology collaboration (CKD-EPI) equation with gold standard method of assessment of GFR by plasma clearance of Tc-99mDTPA is not well-studied in Indian population. We conducted this study to compare GFR estimation by gamma camera-based Gates protocol and serum creatinine-based predicting equations with GFR measured by plasma clearance of Tc-99mDTPA. One hundred and five patients (65 male and 40 female) underwent Tc-99m DTPA renal scan followed by withdrawal of venous blood samples at 2, 3, and 4 h as per predefined protocol. Gates method GFR (GFRs) was assessed using standard protocol. GFR by plasma sampling (GFRp) was calculated by slope-intercept method with provision for corrections. Estimated GFR was calculated by Cockroft-Gault formula, four variable modification of diet in renal disease (MDRD) equation, and CKD-EPI equation (GFRCG, GFRMDRD, GFRCKD-EPI, respectively). GFR measured by gold standard method (GFRp) was compared with that estimated by other methods by calculating correlation coefficient, bias, precision, and accuracy. GFR estimated by all three estimating equations correlated better than GFRs with GFRp. For estimating GFRp, GFRCKD-EPI had highest correlation with GFRp with least bias and highest precision. Gamma camera-based Gates protocol was the least precise and least accurate method for estimating GFRp. To conclude, all three estimating equations based on serum creatinine are superior to Tc-99m DTPA scintigraphy for estimating GFR; CKD-EPI equation being the most accurate and precise.


Multicenter Clinical Evaluation of the Alere i Respiratory Syncytial Virus Isothermal Nucleic Acid Amplification Assay.
The Alere i respiratory syncytial virus (RSV) assay is an isothermal nucleic acid amplification test capable of detecting RSV directly from respiratory specimens, with results being available in ≤13 min after test initiation. The objective of this study was to evaluate the performance characteristics of the Alere i RSV assay in a point-of-care setting by using direct nasopharyngeal (NP) swab specimens (direct NP) and nasopharyngeal swab specimens eluted and transported in viral transport medium (VTM NP). The study was a prospective, multicenter, clinical trial conducted at 9 sites across the United States to evaluate the clinical performance of the Alere i RSV assay with respiratory specimens obtained from both children (age, <18 years) and older adults (age, >60 years). The performance of the Alere i RSV assay was compared with that of the reference method, the Prodesse ProFlu+ real-time reverse transcriptase PCR (RT-PCR) assay. All specimens with discrepant test results were tested further by a second FDA-cleared PCR assay (the Verigene respiratory virus plus nucleic acid test; Luminex Inc., TX). A total of 554 subjects with signs and symptoms of respiratory infections were enrolled, and respiratory samples were collected in this study. In comparison with the ProFlu+ real-time RT-PCR, the overall sensitivity and specificity of Alere i RSV assay for the detection of RSV were 98.6% (95% confidence interval [CI], 94.4 to 99.7%) and 98.0% (95% CI, 95.8 to 99.1%), respectively, for direct NP and 98.6% (95% CI, 94.4 to 99.7%) and 97.8% (95% CI, 95.5 to 98.9%), respectively, for VTM NP. The Alere i RSV is a highly sensitive and specific molecular assay ideal for rapid RSV detection in patients in the point-of-care setting due to its minimal hands-on time and rapid result availability.


Suicidality and risk of suicide--definition, drug safety concerns, and a necessary target for drug development: a brief report.
To address issues concerning potential treatment-emergent "suicidality," a consensus conference was convened March 23-24, 2009. This gathering of participants from academia, government, and industry brought together experts in suicide prevention, clinical trial design, psychometrics, pharmacoepidemiology, and genetics, as well as research psychiatrists involved in studies in studies of psychiatric disorders associated with elevated suicide risk across the life cycle. The process involved reviews of the relevant literature, and a series of 6 breakout sessions focused on specific questions of interest. Each of the participants at the meeting received references relevant to the formal presentations (as well as the slides for the presentations) for their review prior to the meeting. In addition, the assessment instruments of suicidal ideation/behavior were reviewed in relationship to standard measures of validity, reliability, and clinical utility, and these findings were discussed at length in relevant breakout groups, in the final plenary session, and in the preparation of the article. Consensus and dissenting views were noted. Discussion and questions followed each formal presentation during the plenary sessions. Approximately 6 questions per breakout group were prepared in advance by members of the Steering Committee and each breakout group chair. Consensus in the breakout groups was achieved by nominal group process. Consensus recommendations and any dissent were reviewed for each breakout group at the final plenary session. All plenary sessions were recorded and transcribed by a court stenographer. Following the transcript, with input by each of the authors, the final paper went through 14 drafts. The output of the meeting was organized into this brief report and the accompanying full article from which it is distilled. The full article was developed by the authors with feedback from all participants at the meeting and represents a consensus view. Any areas of disagreement at the conference have been noted in the text. The term suicidality is not as clinically useful as more specific terminology (ideation, behavior, attempts, and suicide). Most participants applauded the FDA's encouragement of standard definitions and definable expectations for investigators and industry sponsors. Further research of available assessment instruments is needed to verify their utility, reliability, and validity in identifying suicide-associated treatment-emergent adverse effects and/or a signal of efficacy in suicide prevention trials. The FDA needs to systematically monitor postmarketing events by encouraging the development of a validated instrument for postmarketing surveillance of suicidal ideation, behavior, and risk. Over time, the FDA, industry, and clinical researchers should evaluate the impact of the requirement that all central nervous system clinical drug trials must include a Columbia Classification Algorithm of Suicide Assessment (C-CASA)-compatible screening instrument for assessing and documenting the occurrence of treatment-emergent suicidal ideation and behavior. Finally, patients at high risk for suicide can safely be included in clinical trials, if proper precautions are followed.


Polarizable embedding for simulating redox potentials of biomolecules.
Redox reactions play a key role in various biological processes, including photosynthesis and respiration. Quantitative and predictive computational characterization of redox events is therefore highly desirable for enriching our knowledge on mechanistic features of biological redox-active macromolecules. Here, we present a computational protocol exploiting polarizable embedding hybrid quantum-classical approach and resulting in accurate estimates of redox potentials of biological macromolecules. A special attention is paid to fundamental aspects of the theoretical description such as the effects of environment polarization and of the long-range electrostatic interactions on the computed energetic parameters. Environment (protein and the solvent) polarization is shown to be crucial for accurate estimates of the redox potential: hybrid quantum-classical results with and without account for environment polarization differ by 1.4 V. Long-range electrostatic interactions are shown to contribute significantly to the computed redox potential value even at the distances far beyond the protein outer surface. The approach is tested on simulating reduction potential of cryptochrome 1 protein from Arabidopsis thaliana. The theoretical estimate (0.07 V) of the midpoint reduction potential is in good agreement with available experimental data (-0.15 V).


Xpert® MTB/RIF assay on formalin-fixed paraffin-embedded tissues in the diagnosis of extrapulmonary tuberculosis.
Diagnosis of extrapulmonary tuberculosis continues to be a challenge due to the complexity of the causative organism and the wide array of pathologic features seen in this infection. Xpert MTB/RIF can be used on fresh or frozen tissue specimens for diagnosis of tuberculosis with good results. However, there is little data on its use with formalin-fixed paraffin-embedded (FFPE) tissues. The aim of this study was to demonstrate the potential utility of Xpert MTB/RIF and to compare its performance to Ziehl-Neelsen staining for the detection of Mycobacterium tuberculosis from FFPE tissues using histological features from haematoxylin and eosin staining as the gold standard. Eighty randomly selected archival FFPE tissues exhibiting histological features of tuberculosis were included in the study. After deparaffinisation and lysis, all the tissue specimens were subjected to the Xpert® MTB/RIF assay. The outcome measures were proportions of positively identified cases by each test. Using histology as the gold standard, the sensitivity of Ziehl-Neelsen staining was 20.3% (95% confidence interval: 12% - 30.8%), and the sensitivity of the Xpert® MTB/RIF assay was 53.2% (95% confidence interval: 41.6% - 64.9%); the difference was statistically significant (p = 0.002). None of the cases tested positive for rifampicin resistance. With prior deparaffinisation and lysis, FFPE tissues are amenable to testing by Xpert® MTB/RIF assay. A validation study to determine the clinical utility, analytical optimisation and cost implications of this assay for FFPE tissues is recommended.


"Engaging with birth stories in pregnancy: a hermeneutic phenomenological study of women's experiences across two generations".
The birth story has been widely understood as a crucial source of knowledge about childbirth. What has not been reported is the effect that birth stories may have on primigravid women's understandings of birth. Findings are presented from a qualitative study exploring how two generations of women came to understand birth in the milieu of other's stories. The prior assumption was that birth stories must surely have a positive or negative influence on listeners, steering them towards either medical or midwifery-led models of care. A Heideggerian hermeneutic phenomenological approach was used. Twenty UK participants were purposively selected and interviewed. Findings from the initial sample of 10 women who were pregnant in 2012 indicated that virtual media was a primary source of birth stories. This led to recruitment of a second sample of 10 women who gave birth in the 1970s-1980s, to determine whether they were more able to translate information into knowledge via stories told through personal contact and not through virtual technologies. Findings revealed the experience of 'being-in-the-world' of birth and of stories in that world. From a Heideggerian perspective, the birth story was constructed through 'idle talk' (the taken for granted assumptions of things, which come into being through language). Both oral stories and those told through technology were described as the 'modern birth story'. The first theme 'Stories are difficult like that', examines the birth story as problematic and considers how stories shape meaning. The second 'It's a generational thing', considers how women from two generations came to understand what their experience might be. The third 'Birth in the twilight of certainty,' examines women's experience of Being in a system of birth as constructed, portrayed and sustained in the stories being shared. The women pregnant in 2012 framed their expectations in the language of choice, whilst the women who birthed in the 1970s-1980s framed their experience in the language of safety. For both, however, the world of birth was the same; saturated with, and only legitimised by the birth of a healthy baby. Rather than creating meaningful understanding, the 'idle talk' of birth made both cohorts fearful of leaving the relative comfort of the 'system', and of claiming an alternative birth.


Effects of different organic weed management strategies on the physicochemical, sensory, and antioxidant properties of machine-harvested blackberry fruits.
The effect of 3 different weed management strategies, nonweeding, hand weeding, and weed mat, were examined on physicochemical, sugar profile, and antioxidant properties of 2 cultivars of blackberry (Rubus spp), "Marion" and "Black Diamond" harvested at 3 time intervals during the 2012 season. Sensory analysis on flavor intensity of 6 different descriptors by an experienced panel was also performed on "Black Diamond" berries harvested at the same interval during the 2013 season. While weed management had no effect on pH, titratable acidity, and total soluble solids of either cultivar (P > 0.05), it showed a marked effect on total phenolics (5.65 to 7.80 mg GAE/g FW), total monomeric anthocyanins (1.07 to 2.85 mg/g FW), ORAC (271.51 to 644.97 μMol TE/g FW), FRAP (408.56 to 719.10 μMol Fe(2+) /g FW), sugar profile, and flavor intensity. Hand-weeding resulted in fruit antioxidant content and capacity as much as 30% greater, though the effect was not seen in the late harvest, where the nonweeded samples tended to have higher values. Overall, weed mat samples had the lowest antioxidant content and capacity in all harvests. Sugar profiling exhibited a greater variability based on cultivar and harvest, but overall, weed mat samples had lower sugar levels than fruit from the other 2 methods. Interestingly, the intensity of sensory attributes for "Black Diamond" appear to possibly be inversely related to phenolic and anthocyanin content, with the weed mat management strategy resulting in the highest values for virtually all sensory attributes. This study provided valuable information about the impact of organic production method on the quality of blackberries. Weed management is one of the largest costs associated with organic agriculture because of limited availability of approved herbicides. While much work has been done to evaluate the effect of different methods on plant growth and yield, few have determined the impact of weed management methods on fruit quality. This study investigated the impact of 3 common weed management strategies on physicochemical, sensory, and antioxidant properties of 2 organically grown blackberry cultivars. Given the widespread belief that organically grown products are of higher quality than conventionally grown ones, the information generated is particularly important for growers and consumers.


Global public health impact of recovered supplies from operating rooms: a critical analysis with national implications.
In modern operating rooms, clean and unused medical supplies are routinely discarded and can be effectively recovered and redistributed abroad to alleviate the environmental burden of donor hospitals and to generate substantial health benefits at resource-poor recipient institutions. We established a recovery and donation program to collect clean and unused supplies for healthcare institutions in developing nations. We analyzed items donated over a 3-year period (September 2010-November 2013) by quantity and weight, and estimated the projected value of the program under potential nationwide participation. To capture the health benefits attributable to the donated supplies at recipient institutions, we partnered with two tertiary-care centers in Guayaquil, Ecuador and conducted a pilot study on the utility of the donated supplies at the recipient institutions (October 2013). We determined the disability-adjusted life years (DALY) averted for all patients undergoing procedures involving donated items and estimated the annual attributable DALY as well as the cost per DALY averted both by supply and by procedure. Approximately, 2 million lbs (907,185 kg) per year of medical supplies are recoverable from large non-rural US academic medical centers. Of these supplies, 19 common categories represent a potential for donation worth US $15 million per year, at a cost-utility of US $2.14 per DALY averted. Hospital operating rooms continue to represent a large source of recoverable surgical supplies that have demonstrable health benefits in the recipient communities. Cost-effective recovery and need-based donation programs can significantly alleviate the global burden of surgical diseases.