Linkage disequilibrium in natural populations of Drosophila melanogaster.
Two large, stable populations (Texas and Japan) of Drosophila melanogaster were surveyed at 21 allozyme loci on the second and third chromosomes and for chromosomal gene arrangements on those two chromosomes. Over 220 independent gametes were sampled from each population. The types and frequencies of the surveyed genetic variation are similar to those observed previously and suggest only slight differentiation among geographically distant populations. Linkage disequilibrium among linked allozymes loci is only slightly, if at all, detectable with these sample sizes. Linkage disequilibrium between linked inversions and allozymes loci is common especially when located in the same arm. These disequilibria appear to be in the same direction for most comparisons in the two population samples. This result is interpreted as evidence of similar selective environments (ecological and genetic) in the two populations. It is also noted that the direction of these linkage disequilibria appears to be oriented with respect to the gene frequencies at the component loci.


Improving the Intelligibility of Speech for Simulated Electric and Acoustic Stimulation Using Fully Convolutional Neural Networks.
Combined electric and acoustic stimulation (EAS) has demonstrated better speech recognition than conventional cochlear implant (CI) and yielded satisfactory performance under quiet conditions. However, when noise signals are involved, both the electric signal and the acoustic signal may be distorted, thereby resulting in poor recognition performance. To suppress noise effects, speech enhancement (SE) is a necessary unit in EAS devices. Recently, a time-domain speech enhancement algorithm based on the fully convolutional neural networks (FCN) with a short-time objective intelligibility (STOI)-based objective function (termed FCN(S) in short) has received increasing attention due to its simple structure and effectiveness of restoring clean speech signals from noisy counterparts. With evidence showing the benefits of FCN(S) for normal speech, this study sets out to assess its ability to improve the intelligibility of EAS simulated speech. Objective evaluations and listening tests were conducted to examine the performance of FCN(S) in improving the speech intelligibility of normal and vocoded speech in noisy environments. The experimental results show that, compared with the traditional minimum-mean square-error SE method and the deep denoising autoencoder SE method, FCN(S) can obtain better gain in the speech intelligibility for normal as well as vocoded speech. This study, being the first to evaluate deep learning SE approaches for EAS, confirms that FCN(S) is an effective SE approach that may potentially be integrated into an EAS processor to benefit users in noisy environments.


Association of Patient-Reported Health Status With Long-Term Mortality After Transcatheter Aortic Valve Replacement: Report From the STS/ACC TVT Registry.
Although transcatheter aortic valve replacement (TAVR) is an effective treatment for aortic stenosis, long-term mortality after TAVR remains high and challenging to predict. The Kansas City Cardiomyopathy Questionnaire (KCCQ) is a health status measure, assessed directly from patients, that integrates 2 clinically relevant factors (symptoms and functional status) that may predict TAVR outcomes. Among 7769 patients from 286 sites in the Society of Thoracic Surgeons (STS)/American College of Cardiology (ACC) Transcatheter Valve Therapy (TVT) Registry, we examined the association between preprocedure (baseline) patient health status, as assessed by the KCCQ, and 1-year mortality after TAVR. The KCCQ Overall Summary Score was categorized as very poor: <25, poor: 25 to 49, fair: 50 to 74, or good: ≥75. Before TAVR, health status was rated as very poor in 28%, poor in 38%, fair in 24%, and good in 10%. Patients with worse health status were more likely to be women and had more comorbidities and higher STS mortality risk scores. Compared with those with good health status before TAVR and after adjusting for a broad range of baseline covariates, patients with very poor health status had a 2-fold increased hazard of death over the first year after TAVR (adjusted hazard ratio, 2.00; 95% confidence interval, 1.58-2.54), whereas those with poor and fair health status had intermediate outcomes (adjusted hazard ratio, 1.54; 95% confidence interval, 1.22-1.95 and adjusted hazard ratio, 1.20; 95% confidence interval, 0.94-1.55, respectively). In a national, contemporary practice cohort, worse preprocedure patient health status, as assessed by the KCCQ, was associated with greater long-term mortality after TAVR. These results support the measurement and integration of the KCCQ into mortality risk assessments for patients considering TAVR.


Temporospatial relationship between the expressions of superoxide dismutase and nitric oxide synthase in the developing human brain: immunohistochemical and immunoblotting analyses.
To clarify a significant relationship between superoxide dismutase (SOD) and nitric oxide synthase (NOS) in the developing human brain temporospatially, we demonstrate immunohistochemical expression of Cu/Zn-binding SOD1 (SOD1), Mn-containing SOD2 (SOD2), neuronal NOS (nNOS), inducible NOS (iNOS), and nitrotyrosine in human brains from 13 weeks of gestation to 2 years after birth. The immunoreactivities of both SOD1 and SOD2 were detected in fetal neuroblasts at 13 weeks' gestation, as well as mature neurons at the age of 2 years. By contrast, nNOS neurons could be recognized only at 28 and 33 weeks of gestation in the cerebrum, and only at 15, 18, and 23 weeks of gestation in the brain stem. No significant immunoreactivity for iNOS or nitrotyrosine was detected in any type of cell in any region during any stage examined. Immunoblotting analysis using frontal tissue homogenates at 15, 28, 40 weeks of gestation and 18 months of age revealed single band corresponding to SOD1 molecular weight, observed at all stages examined; a single band compatible with the nNOS molecular mass was detected only at the 28th week of gestation. Together with the fact that nitric oxide (NO) plays a potential role in neuronal differentiation, and that large amounts of NO have cytotoxicity from the reaction of NO with superoxide anions, our data suggested that the expressions of both SOD1 and SOD2, as scavengers of superoxide anions, were maintained from an early developmental stage to prepare stage-specific nNOS expression for a potential differentiation role and to elude NO cytotoxicity.


Tumorigenic and Metastatic Role of CD44-/low/CD24-/low Cells in Luminal Breast Cancer.
Cells with high CD44 but low CD24 expression (CD44high/CD24-/low) and high aldehyde dehydrogenase activity (ALDHbr) are widely considered to be drivers of metastasis, therapy resistance and tumor recurrence in breast cancer. However, the role of the CD44high/CD24-/low and ALDHbr phenotypes in identifying tumorigenic cells in breast cancer remains controversial due to the discrepancy in their distribution and tumorigenic potential in intrinsic breast cancer subtypes. In this study, we analyzed the cells expressing these markers in six different breast cancer cell lines representing major breast cancer subtypes (T47D, MCF-7, BT-474, AU-565, Hs578T and MDA-MB-231). CD44high/CD24-/low, ALDHbr and CD44-/low/CD24-/low cell populations were isolated by flow cytometry and analyzed for hallmark stem cell characteristics of differentiation, migration, invasiveness and metastasis using in vitro and in vivo techniques. Our results demonstrate that the CD44-/low/CD24-/low cell population, which is enriched in luminal cell lines (T47D, MCF-7 and BT-474), possesses metastatic and tumorigenic properties. We also show that, contrary to previous claims, the expression of the ALDH1 isoform ALDH1A1 does not affect the tumorigenic potential of cell lines with high ALDH activity (BT-474 and AU-565). Further transcriptomic and clinical studies are needed to determine the potential of these markers as early diagnostic tools and treatment targets.


Clinical manifestations of primary pulmonary extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue in Japanese population.
We retrospectively analysed 16 cases of newly diagnosed pulmonary mucosa-associated lymphoid tissue (MALT) lymphoma in the Japanese population. The disease was found on the basis of examination findings in 14 cases, and clinical manifestations in 2. According to the extensive staging procedure, four patients had concomitant gastric involvement. Primary treatment involved surgery alone in two patients; surgery followed by rituximab (R)-containing chemotherapy in two; R-containing chemotherapy alone in 11; and chemoradiotherapy without R in one. Over the median observation period of 28 months, disease progression was recorded in three patients, but all 16 patients were alive at the end of the observation period. One patient was treated with R alone and achieved partial remission; subsequent tentative surgery showed no evidence of residual lymphoma. It has been 72 months of progression-free survival after the diagnosis. Primary pulmonary MALT lymphoma exhibited an indolent clinical course. R has potential as a therapeutic agent in patients with pulmonary MALT lymphoma.


Prevalence of morphometric vertebral fractures in patients with type 1 diabetes.
Several studies showed low bone mineral density (BMD) and elevated risk of symptomatic fractures in patients with type 1 diabetes (T1D). To our knowledge, there has been no investigation on the prevalence of asymptomatic vertebral fractures (VFx) in T1D. In the current study, we assessed BMD and the prevalence of VFx in T1D. We evaluated 82 T1D patients (26 males and 56 females, aged 31.1 ± 8.6 years, BMI 23.5 ± 3.3 kg/m(2), disease duration 12.8 ± 8.3 years) and 82 controls (22 females and 60 males, aged 32.9 ± 5.8 years, BMI 23.9 ± 4.8 kg/m(2)). Spinal and femoral BMD (as Z-score, Z-LS and Z-FN, respectively) and the prevalence of VFx were evaluated by dual X-ray absorptiometry. T1D patients had lower Z-LS and Z-FN than controls (-0.55 ± 1.3 vs. 0.35 ± 1.0, P < 0.0001, and -0.64 ± 1.1 vs. 0.29 ± 0.9, P < 0.0001, respectively) and a higher prevalence of VFx (24.4 vs. 6.1%, P = 0.002). Age, diabetes duration, age at diabetes manifestation, glycosylated hemoglobin, Z-LS, Z-FN, and the prevalence of chronic complications were similar for patients with and without VFx. In the logistic regression analysis, the presence of VFx was associated with the presence of T1D but not with lumbar spine BMD. Whereas moderate or severe VFx was associated with low lumbar spine BMD in the whole combined group of T1D patients and controls, there was no association between moderate or severe VFx and lumbar spine BMD in the T1D group. T1D patients have low BMD and elevated prevalence of asymptomatic VFx, which is associated with the presence of T1D independently of BMD.


Structural diversity of angiotensin-converting enzyme.
The crystal structure of a Drosophila angiotensin-converting enzyme (ANCE) has recently been solved, revealing features important for the binding of ACE inhibitors and allowing molecular comparisons with the structure of human testicular angiotensin-converting enzyme (tACE). ACER is a second Drosophila ACE that displays both common and distinctive properties. Here we report further functional differences between ANCE and ACER and have constructed a homology model of ACER to help explain these. The model predicts a lack of the Cl(-)-binding sites, and therefore the strong activation of ACER activity towards enkephalinamide peptides by NaCl suggests alternative sites for Cl(-) binding. There is a marked difference in the electrostatic charge of the substrate channel between ANCE and ACER, which may explain why the electropositive peptide, MKRSRGPSPRR, is cleaved efficiently by ANCE with a low K(m), but does not bind to ACER. Bradykinin (BK) peptides are excellent ANCE substrates. Models of BK docked in the substrate channel suggest that the peptide adopts an N-terminal beta-turn, permitting a tight fit of the peptide in the substrate channel. This, together with ionic interactions between the guanidino group of Arg9 of BK and the side chains of Asp360 and Glu150 in the S(2)' pocket, are possible reasons for the high-affinity binding of BK. The replacement of Asp360 with a histidine in ACER would explain the higher K(m) recorded for the hydrolysis of BK peptides by this enzyme. Other differences in the S(2)' site of ANCE and ACER also explain the selectivity of RXPA380, a selective inhibitor of human C-domain ACE, which also preferentially inhibits ACER. These structural and enzymatic studies provide insight into the molecular basis for the distinctive enzymatic features of ANCE and ACER.


Unified prebiotically plausible synthesis of pyrimidine and purine RNA ribonucleotides.
Theories about the origin of life require chemical pathways that allow formation of life's key building blocks under prebiotically plausible conditions. Complex molecules like RNA must have originated from small molecules whose reactivity was guided by physico-chemical processes. RNA is constructed from purine and pyrimidine nucleosides, both of which are required for accurate information transfer, and thus Darwinian evolution. Separate pathways to purines and pyrimidines have been reported, but their concurrent syntheses remain a challenge. We report the synthesis of the pyrimidine nucleosides from small molecules and ribose, driven solely by wet-dry cycles. In the presence of phosphate-containing minerals, 5'-mono- and diphosphates also form selectively in one-pot reactions. The pathway is compatible with purine synthesis, allowing the concurrent formation of all Watson-Crick bases.


Predicting program start-up using the stages of implementation measure.
Recent efforts to better understand the process of implementation have been hampered by a lack of tools available to define and measure implementation progress. The Stages of Implementation Completion (SIC) was developed as part of an implementation trial of MTFC in 53 sites, and identifies the duration of time spent on implementation activities and the proportion of activities completed. This article examines the ability of the first three stages of the SIC (Engagement, Consideration of Feasibility, Readiness Planning) to predict successful program start-up. Results suggest that completing SIC stages completely, yet relatively quickly, predicts the likelihood of successful implementation.


Mental and Social Health Impacts the Use of Protective Behavioral Strategies in Reducing Risky Drinking and Alcohol Consequences.
The present study is the first to examine the moderating effects of mental and social health status in the relationship between protective behavioral strategies utilized to reduce high-risk drinking (e.g., alternating alcoholic and nonalcoholic drinks or avoiding drinking games) and alcohol outcomes (drinking variables and alcohol-related negative consequences) among first-year college females (N = 128). Findings revealed that protective behaviors were particularly effective in reducing both alcohol consumption and related risks among participants reporting lower mental health as compared to higher mental health. Further, participants with higher social health who utilized protective behaviors consumed significantly fewer maximum drinks per occasion than did peers who also employed protective behaviors but reported lower social health. Explanation of findings and implications for campus intervention initiatives are discussed.


Heterozygous TGFBR2 mutations in Marfan syndrome.
Marfan syndrome is an extracellular matrix disorder with cardinal manifestations in the eye, skeleton and cardiovascular systems associated with defects in the gene encoding fibrillin (FBN1) at 15q21.1 (ref. 1). A second type of the disorder (Marfan syndrome type 2; OMIM 154705) is associated with a second locus, MFS2, at 3p25-p24.2 in a large French family (family MS1). Identification of a 3p24.1 chromosomal breakpoint disrupting the gene encoding TGF-beta receptor 2 (TGFBR2) in a Japanese individual with Marfan syndrome led us to consider TGFBR2 as the gene underlying association with Marfan syndrome at the MSF2 locus. The mutation 1524G-->A in TGFBR2 (causing the synonymous amino acid substitution Q508Q) resulted in abnormal splicing and segregated with MFS2 in family MS1. We identified three other missense mutations in four unrelated probands, which led to loss of function of TGF-beta signaling activity on extracellular matrix formation. These results show that heterozygous mutations in TGFBR2, a putative tumor-suppressor gene implicated in several malignancies, are also associated with inherited connective-tissue disorders.


Where we are in the fight against Hepatitis B Infection; Trends in Hepatitis B virus seroprevalence in Black Sea Region of Turkey.
To determine new strategies for complete coverage of hepatitis B virus (HBV) vaccination, every country needs to take into concern factors of infection transmission in its own region. The aim of this study was to investigate the seroprevalence of hepatitis B among all age groups in northern Turkey using HBsAg and anti-HBs serological markers. The laboratory records of a total of 101648 patients of all ages attending a tertiary level hospital in Samsun, a Black Sea coastal city, between January 2014 and May 2016 were evaluated retrospectively. HBsAg and anti-HBs seropositivity was found to be 4% and 38.3%, respectively. There was a significant difference between HBsAg (χ2 = 209.08; P = 0.00), anti-HBs (χ2 = 124.12; P = 0.00) seropositivity, and immunization status. Although we found a statically difference between men and women (χ2 = 32.2 P = 0.00) for HBsAg seropositivity, there was no significant difference for anti-HBs (P = 0.22). In 1998, the universal infant immunization program changed the HBV epidemiology in Turkey, and resulted in an apparent trend towards reduced disease levels. However, prevalence of HBV infection is still high in adolescent and young adults. Therefore, a catch-up immunization program, education, and follow-up policy for these groups, in addition to routine infant immunization, will decrease the HBV infection rate, reducing morbidity and mortality rates, and will help to reduce hepatitis B transmission in Turkey.


A Southwestern Tribal Perspective on Traditional and Commercial Tobacco.
American Indian or Alaska Natives have the highest rates of current cigarette (36.5%) and smokeless tobacco use (5.3%), and tobacco product (40.1%) and the second highest rate of current cigar use (6.1%) compared to all other racial-ethnic groups in the U.S. rates of American Indian or Alaska Native tobacco use vary by gender. Few studies examine perceptions of tobacco use among tribal members residing on and off the reservation. This study fills a gap in the literature by reporting the perceptions of 34 enrolled members of a southwestern tribe who reside on and off a tribal land using a Community-Based Participatory Research (CBPR) design through a collaboration between a university and a tribal health program. Researchers conducted seven focus groups; four on the southwest reservation and three within an urban community. The discussions were audio-recorded, transcribed, and analyzed using a multi-investigator consensus model. The use of tobacco (commercial or traditional) in southwest tribes is essential to cultural practices. Results depicted different views on cultural meaning and health impacts of commercial and traditional tobacco. Findings suggest the importance of local research to understand dimensions of tobacco use before moving forward with tobacco cessation programming.


Propagation velocity kinetics and repolarization alternans in a free-behaving sheep model of pacing-induced atrial fibrillation.
Experimental models have reported conflicting results regarding the role of dispersion of repolarization in promoting atrial fibrillation (AF). Repolarization alternans, a beat-to-beat alternation in action potential duration, enhances dispersion of repolarization when propagation velocity is involved. In this work, original electrophysiological parameters were analysed to study AF susceptibility in a chronic sheep model of pacing-induced AF. Two pacemakers were implanted, each with a single right atrial lead. Right atrial depolarization and repolarization waves were documented at 2-week intervals. A significant and gradual decrease in the propagation velocity at all pacing rates and in the right atrial effective refractory period (ERP) was observed during the weeks of burst pacing before sustained AF developed when compared with baseline conditions. Right atrial repolarization alternans was observed, but because of the development of 2/1 atrioventricular block with far-field ventricular interference, its threshold could not be precisely measured. Non-sustained AF was not observed at baseline, but appeared during the electrical remodelling in association with a decrease in both ERP and propagation velocity. We report here on the feasibility of measuring ERP, atrial repolarization alternans, and propagation velocity kinetics and their potential in predicting susceptibility to AF in a free-behaving model of pacing-induced AF using the standard pacemaker technology.


Dietary fat and fiber in relation to risk of breast cancer. An 8-year follow-up.
To address the hypotheses that dietary fat increases and fiber decreases the risk of breast cancer. Prospective cohort study with dietary assessment at baseline, using a validated, self-administered food frequency questionnaire. 89,494 women in the Nurses' Health Study who were 34 through 59 years of age in 1980 and who were followed up for 8 years (> 95% complete). 1439 incident cases of breast cancer were diagnosed, including 774 among postmenopausal women. After adjustment for age, established risk factors, and total energy intake, we observed no evidence of any positive association between total fat intake and breast cancer incidence (relative risks [RRs] for increasing quintiles of fat intake were 1.0, 0.85, 0.96, 0.91, and 0.90; 95% confidence interval for highest vs lowest quintile, 0.77 to 1.07). Among postmenopausal women alone, corresponding RRs were 1.0, 0.89, 1.00, 0.95, and 0.91. Comparing extreme deciles of total fat intake (> or = 49% vs < 29% of total energy intake), the RR was 0.86 (95% confidence interval, 0.67 to 1.08). A similar absence of any positive association was observed without adjustment for energy intake; for tumors less than 2 cm as well as 2 cm or greater in diameter; for saturated, monounsaturated, and polyunsaturated fat; and after excluding the first 4 years of follow-up. Also, we found no suggestion of any positive association when using a more detailed and precise dietary questionnaire completed in 1984 (666 subsequent cases), even when women consuming less than 25% of energy from fat were used as the comparison group. No suggestion of a protective effect of dietary fiber was observed (RRs for increasing quintiles were 1.0, 0.95, 0.93, 1.02, and 1.02). These data provide evidence against both an adverse influence of fat intake and a protective effect of fiber consumption by middle-aged women on breast cancer incidence over 8 years. Nevertheless, the positive association between intake of animal fat and risk of colon cancer observed in many studies provides ample reason to limit this source of energy.


High incidence of abnormal glucose metabolism in acute coronary syndrome patients at a moderate altitude: A sub-Himalayan study.
Abnormal glucose metabolic status at admission is an important marker of future cardiovascular events and long-term mortality after acute coronary syndrome (ACS), whether or not they are known diabetics. The aims were to study the prevalence of abnormal glucose metabolism in ACS patients and to compare the different methods of diagnosing diabetes in ACS patients. We did a prospective study. About 250 consecutive nondiabetic patients (200 men and 50 women) with ACS admitted to a tertiary care institute of Himachal Pradesh in 1 year were enrolled. Admission plasma glucose, next morning fasting plasma glucose (FPG), A1C, and a standardized 75-g oral glucose tolerance test (OGTT) 72 h after admission were done. Glucose metabolism was categorized as normal glucose metabolism, impaired glucose metabolism (impaired fasting glucose or impaired glucose tolerance [IGT]), and diabetes. Diabetes was arbitrarily classified further as undiagnosed (HBA1c ≥6.5%) or possibly stress diabetes (HBA1c <6.5%). A repeat OGTT after 3 months in objects with IGT and stress hyperglycemia at a time of admission was done. The mean age was 54 ± 12.46 years. The mean plasma glucose at admission was 124 ± 53.96 mg/dL, and the mean FPG was 102 ± 27.07 mg/dL. The mean 2-h postglucose load concentration was 159.5 ± 56.58 mg/dL. At baseline, 95 (38%) had normal glucose metabolism, 95 (38%) had impaired glucose metabolism (IGT and or IGT) and 60 (24%) had diabetes; 48 (19.2%) were undiagnosed diabetes and 12 (4.8%) had stress hyperglycemia. At follow up 58.66% and 55.55% of patients with impaired glucose tolerance and stress hyperglycemia continued to have impaired glucose tolerance respectively. About 75 gm OGTT has highest sensitivity and specificity to diagnose diabetes, whereas A1C most specific to rule out stress hyperglycemia. In this small hilly state of India, abnormal glucose metabolism (previously undiagnosed diabetes and IGT) is common in patients admitted with ACS. Abnormal glucometabolic status can be detected early in the postadmission period. Our results further suggest that 75-g OGTT remained the gold standard test to detect diabetes and could be used before discharge to diagnose diabetes.


Glucose-induced glycogenesis in the liver involves the glucose-6-phosphate-dependent dephosphorylation of glycogen synthase.
Non-metabolized glucose derivatives may cause inactivation of phosphorylase but, unlike glucose, they are unable to elicit activation of glycogen synthase in isolated hepatocytes. We report here that, after the previous inactivation of phosphorylase by one of these glucose derivatives (2-deoxy-2-fluoro-alpha-glucosyl fluoride), glycogen synthase was progressively activated by addition of increasing concentrations of glucose. Under these conditions, the degree of activation of glycogen synthase was linearly correlated with the intracellular glucose-6-phosphate (Glc-6-P) concentration. Addition of glucosamine, an inhibitor of glucokinase, decreased both parameters in parallel. Further experiments using an inhibitor of either protein kinases (5-iodotubercidin) or protein phosphatases (microcystin) in isolated hepatocytes indicated that Glc-6-P does not affect glycogen-synthase kinase activity but enhances the glycogen-synthase phosphatase reaction. Experiments in vitro showed that the synthase phosphatase activity of glycogen-bound type-1 protein phosphatase was increased by physiological concentrations of Glc-6-P (0.1-0.5 mM), but not by 2.5 mM fructose-6-P, fructose-1-P or glucose-1-P. At physiological ionic strength, the glycogen-associated synthase phosphatase activity was nearly entirely Glc-6-P-dependent, but Glc-6-P did not relieve the strong inhibitory effect of phosphorylase a. The large stimulatory effects of 2.5 mM Glc-6-P, with glycogen synthase b and phosphorylase a as substrates, appeared to be mostly substrate-directed, while the modest effects observed with casein and histone IIA pointed to an additional stimulation of glycogen-bound protein phosphatase-1 by Glc-6-P. We conclude that glucose elicits hepatic synthase phosphatase activity both by removal of the inhibitor, phosphorylase a, and by generation of the stimulator, Glc-6-P.


Heroin vs morphine for cancer pain?
Narcotic analgesics are the mainstay of pain control in patients with cancer. A controversy has been raging in the United States and Canada as to the legalization of heroin. We have reviewed the literature in order to determine the relative efficacy of heroin and morphine in cancer pain. We applied the following methodologic criteria: Was the assignment of patients to the different opiates randomized? Were all clinically relevant outcomes reported? Were the patients recognizable? Were both clinical and statistical significance considered? Was the opiate regimen feasible in routine clinical practice? Were all patients who entered the study accounted for at its conclusion? Two trials satisfied our first standard. The first, a double-blind cross-over trial, failed to meet standard 4 (the negative conclusion may represent a type 2 error) and only 21% of patients completed both treatment periods. The second study, which compared intramuscular heroin and morphine among patients with postoperative pain, failed to meet standards 3 (patients not described in sufficient detail and only tangentially related to chronic cancer pain) and 4 (type 2 error). Thus the relative efficacy of heroin and morphine in the relief of chronic cancer pain remains unknown. Randomized trials that meet all six methodologic standards must therefore be carried out for this controversy to be resolved.


Quantification Analysis of Sleep Based on Smartwatch Sensors for Parkinson's Disease.
Rapid eye movement (REM) sleep behavior disorder (RBD) is associated with Parkinson's disease (PD). In this study, a smartwatch-based sensor is utilized as a convenient tool to detect the abnormal RBD phenomenon in PD patients. Instead, a questionnaire with sleep quality assessment and sleep physiological indices, such as sleep stage, activity level, and heart rate, were measured in the smartwatch sensors. Therefore, this device can record comprehensive sleep physiological data, offering several advantages such as ubiquity, long-term monitoring, and wearable convenience. In addition, it can provide the clinical doctor with sufficient information on the patient's sleeping patterns with individualized treatment. In this study, a three-stage sleep staging method (i.e., comprising sleep/awake detection, sleep-stage detection, and REM-stage detection) based on an accelerometer and heart-rate data is implemented using machine learning (ML) techniques. The ML-based algorithms used here for sleep/awake detection, sleep-stage detection, and REM-stage detection were a Cole-Kripke algorithm, a stepwise clustering algorithm, and a k-means clustering algorithm with predefined criteria, respectively. The sleep staging method was validated in a clinical trial. The results showed a statistically significant difference in the percentage of abnormal REM between the control group (1.6 ± 1.3; n = 18) and the PD group (3.8 ± 5.0; n = 20) (p = 0.04). The percentage of deep sleep stage in our results presented a significant difference between the control group (38.1 ± 24.3; n = 18) and PD group (22.0 ± 15.0, n = 20) (p = 0.011) as well. Further, our results suggested that the smartwatch-based sensor was able to detect the difference of an abnormal REM percentage in the control group (1.6 ± 1.3; n = 18), PD patient with clonazepam (2.0 ± 1.7; n = 10), and without clonazepam (5.7 ± 7.1; n = 10) (p = 0.007). Our results confirmed the effectiveness of our sensor in investigating the sleep stage in PD patients. The sensor also successfully determined the effect of clonazepam on reducing abnormal REM in PD patients. In conclusion, our smartwatch sensor is a convenient and effective tool for sleep quantification analysis in PD patients.


On silico peptide microarrays for high-resolution mapping of antibody epitopes and diverse protein-protein interactions.
We developed a new, silicon-based peptide array for a broad range of biological applications, including potential development as a real-time point-of-care platform. We used photolithography on silicon wafers to synthesize microarrays (Intel arrays) that contained every possible overlapping peptide within a linear protein sequence covering the N-terminal tail of human histone H2B. These arrays also included peptides with acetylated and methylated lysine residues, reflecting post-translational modifications of H2B. We defined minimum binding epitopes for commercial antibodies recognizing the modified and unmodified H2B peptides. We further found that this platform is suitable for the highly sensitive characterization of methyltransferases and kinase substrates. The Intel arrays also revealed specific H2B epitopes that are recognized by autoantibodies in individuals with systemic lupus erythematosus who have elevated disease severity. By combining emerging nonfluorescence-based detection methods with an underlying integrated circuit, we are now poised to create a truly transformative proteomics platform with applications in bioscience, drug development and clinical diagnostics.


Drug therapy for the treatment of chronic nonspecific low back pain: systematic review and meta-analysis.
Low back pain (LBP) is one of the most common health problems in adults. The impact of LBP on the individual can cause loss of health status in the form of symptoms and loss of function related to pain in the back; limitation of daily, leisure, and/or strenuous activities, and disability. LBP also poses an economic burden to society, mainly in terms of one of the most common reasons for seeking medical care (direct treatment costs), and accounts for the large number of work days lost (indirect costs). To reduce the impact of LBP on adults, drug therapy is the most frequently recommended intervention. Over the last decade, a substantial number of randomized clinical trials of drug therapy for LBP have been published. To determine the effectiveness of drug therapy for the treatment of chronic nonspecific low back pain (CNLBP). Systematic review A systematic review and meta-analysis of randomized controlled trials was conducted. Five databases (Medline, CINAHL, Science Direct, CAJ Full-text Database, and Cochrane databases) were searched for articles published from 2002 to 2012. The eligibility criteria were randomized trials and double-blind controlled trials of oral or injection drug therapy for CNLBP in subjects who were aged at least 18 years old, published in English or Chinese. Two independent reviewers extracted the data. A total of 25 drug therapy trials were included. cyclo-oxygenase-2 (COX-2) nonsteroidal anti-inflammatory drugs (NSAIDs), tramadol, and opioids were commonly used. Only 5 trials studied the efficacy of adjuvant analgesics of antiepileptics (n = 1) and antidepressants (n = 4) for CNLBP. The standardized mean difference (SMD) for COX-2 NSAIDs in pain relief was -12.03 (95% confidence interval [CI]: -15.00 to -9.06). The SMD for tramadol in pain relief was -1.72 (95% CI: -3.45 to 0.01). As the 95% CI crossed 0, this effect size was not considered statistically significant. The SMD for the overall effects of opioids in pain relief was -5.18 (95% CI: -8.30 to -2.05). The SMD for the partial opioid agonist drug in pain relief was -7.46 (95% CI: -11.87 to -3.04). The follow-up periods of these included trials in the meta-analysis ranged from 4 to 24 weeks. The difference of follow-up periods influenced how study outcomes were recorded. These included trials also had significant differences in patient selections. Some trials may actually include CNLBP patients with neuropathic pain, as not having focal neurological findings or signs does not mean that the pain is not neuropathic. Consequently, different pain conditions may influence patients who responded to the same drug and then influence pooled estimates of treatment effect size. This review endorses the use of COX-2 NSAIDs as the first-line drugs for CNLBP. Tramadol shows no statistically significant effect on pain relief, but has small effect sizes in improving functioning. Among included opioid therapy studies, the overall effects of opioids and the partial opioids agonist drug had statistically significant treatment effects in pain relief for CNLBP patients.


Recombinant factor VIIa and the surgical patient.
Bleeding remains a challenge in surgery. A unique drug, recombinant factor VIIa, causes clotting exclusively at bleeding sites. Recombinant factor VIIa has recently been introduced to surgery where current evidence, consisting mostly of case reports, suggest remarkable safety and efficacy. The first randomized controlled trials are only now being published with less remarkable results. This manuscript summarizes the current evidence. In trauma, a single randomized control trial suggests recombinant factor VIIa reduces bleeding and transfusion in blunt trauma, particularly in coagulopathic patients. In cardiac surgery, one randomized control trial, open-label studies and case reports suggest benefit in refractory bleeding. For liver surgery, randomized control trials do not support use in liver transplant or gastrointestinal bleeding. In neurosurgery, one randomized control trial demonstrated improved outcome in intracerebral hemorrhage. In urology, one randomized control trial demonstrated significant reduction in perioperative bleeding. For orthopedics, a single randomized control trial showed no benefit in pelvic/acetabular surgery. In obstetrics/gynecology, limited evidence suggests benefit in massive bleedings. Current evidence does not yet support recombinant factor VIIa as standard of care in surgery. However, the evidence indicates that recombinant factor VIIa should be used in intracerebral hemorrhage and massive perioperative or traumatic bleeding refractory to conventional therapies. For now, the bedside decision to use recombinant factor VIIa remains a matter of surgical judgment.


Innate Inhibiting Proteins Enhance Expression and Immunogenicity of Self-Amplifying RNA.
Self-amplifying RNA (saRNA) is a cutting-edge platform for both nucleic acid vaccines and therapeutics. saRNA is self-adjuvanting, as it activates types I and III interferon (IFN), which enhances the immunogenicity of RNA vaccines but can also lead to inhibition of translation. In this study, we screened a library of saRNA constructs with cis-encoded innate inhibiting proteins (IIPs) and determined the effect on protein expression and immunogenicity. We observed that the PIV-5 V and Middle East respiratory syndrome coronavirus (MERS-CoV) ORF4a proteins enhance protein expression 100- to 500-fold in vitro in IFN-competent HeLa and MRC5 cells. We found that the MERS-CoV ORF4a protein partially abates dose nonlinearity in vivo, and that ruxolitinib, a potent Janus kinase (JAK)/signal transducer and activator of transcription (STAT) inhibitor, but not the IIPs, enhances protein expression of saRNA in vivo. Both the PIV-5 V and MERS-CoV ORF4a proteins were found to enhance the percentage of resident cells in human skin explants expressing saRNA and completely rescued dose nonlinearity of saRNA. Finally, we observed that the MERS-CoV ORF4a increased the rabies virus (RABV)-specific immunoglobulin G (IgG) titer and neutralization half-maximal inhibitory concentration (IC50) by ∼10-fold in rabbits, but not in mice or rats. These experiments provide a proof of concept that IIPs can be directly encoded into saRNA vectors and effectively abate the nonlinear dose dependency and enhance immunogenicity.


Multi-tissue coexpression networks reveal unexpected subnetworks associated with disease.
Obesity is a particularly complex disease that at least partially involves genetic and environmental perturbations to gene-networks connecting the hypothalamus and several metabolic tissues, resulting in an energy imbalance at the systems level. To provide an inter-tissue view of obesity with respect to molecular states that are associated with physiological states, we developed a framework for constructing tissue-to-tissue coexpression networks between genes in the hypothalamus, liver or adipose tissue. These networks have a scale-free architecture and are strikingly independent of gene-gene coexpression networks that are constructed from more standard analyses of single tissues. This is the first systematic effort to study inter-tissue relationships and highlights genes in the hypothalamus that act as information relays in the control of peripheral tissues in obese mice. The subnetworks identified as specific to tissue-to-tissue interactions are enriched in genes that have obesity-relevant biological functions such as circadian rhythm, energy balance, stress response, or immune response. Tissue-to-tissue networks enable the identification of disease-specific genes that respond to changes induced by different tissues and they also provide unique details regarding candidate genes for obesity that are identified in genome-wide association studies. Identifying such genes from single tissue analyses would be difficult or impossible.


A view from the streets: women and medical work in Elizabethan London.
In Elizabethan London, women occupied a significant position in the city's medical marketplace, both as consumers of medical services and as practitioners. Though male medical authors of the period objected to the presence and practices of these women, a very different view of their medical work emerges if we shift our historical vantage point to the streets, houses, churches, and hospitals of the city. Using relatively underutilized sources such as parish records, probate records, lists of immigrants to London, hospital records, and individual manuscripts it is possible to draw a richer, more detailed portrait of how female health-care workers engaged with the business of health and healing. Women emerge from these records as active, prominent, and acknowledged participants in the delivery of services that promoted and preserved the health of many Londoners from cradle to grave. Hired by public institutions such as parishes and hospitals, as well as by private individuals, women were central figures in the delivery of nursing, medical, pharmaceutical, and surgical services throughout the city as part of organized systems of health care. Exploring how Londoners saw female practitioners, and how women played a recognized role within the city's range of health-care options, demonstrates that women were crucial to community health, and were also valued as such by their neighbors and patients.


Identifying clusters of falls-related hospital admissions to inform population targets for prioritising falls prevention programmes.
There has been limited research investigating the relationship between injurious falls and hospital resource use. The aims of this study were to identify clusters of community-dwelling older people in the general population who are at increased risk of being admitted to hospital following a fall and how those clusters differed in their use of hospital resources. Analysis of routinely collected hospital admissions data relating to 45 374 fall-related admissions in Victorian community-dwelling older adults aged ≥65 years that occurred during 2008/2009 to 2010/2011. Fall-related admission episodes were identified based on being admitted from a private residence to hospital with a principal diagnosis of injury (International Classification of Diseases (ICD)-10-AM codes S00 to T75) and having a first external cause of a fall (ICD-10-AM codes W00 to W19). A cluster analysis was performed to identify homogeneous groups using demographic details of patients and information on the presence of comorbidities. Hospital length of stay (LOS) was compared across clusters using competing risks regression. Clusters based on area of residence, demographic factors (age, gender, marital status, country of birth) and the presence of comorbidities were identified. Clusters representing hospitalised fallers with comorbidities were associated with longer LOS compared with other cluster groups. Clusters delineated by demographic factors were also associated with increased LOS. All patients with comorbidity, and older women without comorbidities, stay in hospital longer following a fall and hence consume a disproportionate share of hospital resources. These findings have important implications for the targeting of falls prevention interventions for community-dwelling older people.


The effect of sequence selection and field strength on detection of osteochondral defects in the metacarpophalangeal joint.
Six cadaver forelimbs were imaged in two high-field magnetic resonance (MR) systems and one low-field MR system following the creation of osteochondral defects on the palmar distal aspect of the third metacarpal bone. The following sequences were performed using all three systems: proton density (PD) turbo spin echo, T2* gradient echo (GRE), T2-weighted fast spin echo, and short tau inversion recovery. In addition, 3D T1 GRE sagittal standard and motion insensitive sequences were obtained using the low-field system. PD fat saturated and 3D T1-weighted spoiled GRE images with and without fat suppression were acquired with the high-field systems. Lesions were measured and assigned a confidence score. The images obtained using high-field systems (1.0 and 1.5 T) more accurately represented the osteochondral defects when compared with low-field system (0.27 T) images. The largest difference was observed when evaluating articular cartilage defects, which were not identified on the low-field images. Sequence selection affected the appearance of the lesions. On all systems the turbo and fast spin echo sequences more accurately represented the lesion size and shape when compared with the GRE sequences. The T1 GRE sequence is the only sequence that appears to allow visualization of the articular cartilage on the low-field images, but is limited in providing adequate cartilage visualization. Confidence scores were greater on the high-field systems when compared with the low-field system.


Prediction of Opioid Analgesic Efficacy by Measurement of Pupillary Unrest.
Pupillary unrest under ambient light (PUAL) is the fluctuation in pupil diameter in time around a mean value. PUAL is augmented by light and diminished by administration of opioids. We hypothesized that, because pupillary unrest is a marker of opioid effect, low levels of PUAL may be associated with reduced opioid efficacy, as measured by changes in the numerical rating scale (NRS) pain scores of patients in the postanesthesia care unit (PACU). We used an infrared pupillometer to measure PUAL in patients recovering from ambulatory surgery at 2 different institutions. At both sites, PUAL was quantified using spectral analysis of the Fourier transform of pupil diameter versus time. We measured PUAL and pain scores before and after opioid administration. Protocols for total capture time and lighting conditions varied between the 2 sites. Correlations between PUAL and change in NRS scores were examined using significance testing of Pearson correlation coefficients. Correlations between change in PUAL and change in NRS scores were also examined. Patients were divided into high and low PUAL groups, and high and low response to opioid. A Fisher exact test was used to determine whether there was a significant association between PUAL and opioid response. For patients with pain in the PACU, low levels of pupillary unrest before opioid therapy were associated with minimal or no reduction in pain scores after opioid administration. We noted a significant correlation at both sites between PUAL and pain score reduction with opioids (r = 0.59, P = .0053, and r = 0.57, P = .022.) The Fisher exact test confirmed that patients with PUAL levels above the mean had a more beneficial analgesic effect from opioids than those with low PUAL levels (P = .018). We also noted that change in PUAL was significantly correlated with change in pain score at both sites (r = 0.56, P = .03 and r = 0.55, P = .01). We observe that the pretreatment magnitude of PUAL is correlated with the analgesic response to opioid therapy, and that patients who exhibit higher levels of PUAL change after opioid administration have a more beneficial analgesic effect from opioids. Larger studies with uniform measurement protocols are required to confirm these preliminary results.


Decorin Prevents Retinal Pigment Epithelial Barrier Breakdown Under Diabetic Conditions by Suppressing p38 MAPK Activation.
The purpose of this study was to determine the effect of decorin on the barrier function of human retinal pigment epithelial (RPE) cells under high-glucose (HG) plus hypoxia conditions. Human RPE (ARPE-19) cells were cultured for 18 days in normal glucose (5.5 mM) or HG (25 mM) medium. In addition, to mimic the hypoxic impact which occurs in diabetic retinopathy, cells were treated with 100 μM CoCl2 during the last 2 days of the experiment. Decorin, 100 nM, was applied 1 hour before CoCl2 was added. Retinal pigment epithelial barrier function was evaluated by measuring transepithelial electrical resistance (TER) and apical-basolateral permeability of fluorescein isothiocyanate (FITC)-dextran. The content and distribution of tight junction proteins (claudin-1, occludin, and zonula occludens-1 [ZO-1]) were examined by Western blotting and immunofluorescence. p38 mitogen-activated protein kinase (MAPK) phosphorylation was evaluated by Western blotting, and small interfering RNA transfection to p38 MAPK was also performed in ARPE-19 monolayers. High-glucose plus hypoxia significantly increased FITC-dextran permeability, paralleled by decreased TER. Decorin reversed both of these effects. High-glucose plus hypoxia-induced reduction and disorganization of occludin and ZO-1 were also reversed by decorin. Decorin prevented the activation of p38 MAPK induced by hypoxia. Silence of p38 MAPK by RNA interference also inhibited the breakdown of ARPE-19 cell monolayer induced by HG plus hypoxia. Retinal pigment epithelial barrier disruption induced by HG plus hypoxia was prevented by decorin through suppression of p38 MAPK activation, which could present a new therapeutic strategy for inhibition of diabetic macular edema.


Bronchoscopic lung volume reduction: recent updates.
Emphysema causes significant morbidity and mortality, incurring both financial and psychosocial costs. Alternatives to medical therapy and surgical lung volume reduction surgery (LVRS) have increased interest in bronchoscopic techniques. Bronchoscopic lung volume reduction (BLVR) is still in its infancy and additional trials and follow-up are critical. However, several new randomized clinical trials (RCTs) have demonstrated improvement in lung function, quality of life and exercise capacity in select patients receiving endobronchial valves and coil therapy. This article highlights recent data regarding bronchoscopic treatment of emphysema.


An approximate short scan helical FDK cone beam algorithm based on nutating curved surfaces satisfying the Tuy's condition.
Traditionally, short scan helical FDK algorithms have been implemented based on horizontal transaxial slices. However, not every point on the horizontal transaxial slice satisfies Tuy's condition for the corresponding (pi+fan angle) segment of helix, which means that some points on the horizontal slices are incompletely sampled and are impossible to be exactly reconstructed. In this paper, we propose and implement an improved but still approximate short scan helical cone beam FDK algorithm based on nutating curved surfaces satisfying the Tuy's condition. This surface is defined by averaging PI surfaces emanating the initial and final source points of a (pi+fan angle) segment of helix. One of the key characteristics of the surface is that every point on it satisfies the Tuy's condition for the corresponding (pi+fan angle) segment of helix, which means that we can potentially reconstruct every point on the surface exactly. This difference makes the proposed algorithm deliver a better-reconstructed image quality while requiring a smaller detector area than that of traditional FDK methods based on horizontal transaxial slices. Another characteristic of the proposed surface is that every point within the helix belongs to one and only one such surface. Therefore, the location of the short scan segment for the reconstruction of a point in Cartesian coordinate could be precalculated and stored in a look-up table. This enables us to perform reconstruction directly on rectangular grids. We compare the performance of the improved FDK algorithm with that of a quasi-exact algorithm based on data combination technique. The simulation results show that the reconstructed image quality of these two methods is similar.


Biochemical and Structural Basis for Controlling Chemical Modularity in Fungal Polyketide Biosynthesis.
Modular collaboration between iterative fungal polyketide synthases (IPKSs) is an important mechanism for generating structural diversity of polyketide natural products. Inter-PKS communication and substrate channeling are controlled in large by the starter unit acyl carrier protein transacylase (SAT) domain found in the accepting IPKS module. Here, we reconstituted the modular biosynthesis of the benzaldehyde core of the chaetoviridin and chaetomugilin azaphilone natural products using the IPKSs CazF and CazM. Our studies revealed a critical role of CazM's SAT domain in selectively transferring a highly reduced triketide product from CazF. In contrast, a more oxidized triketide that is also produced by CazF and required in later stages of biosynthesis of the final product is not recognized by the SAT domain. The structural basis for the acyl unit selectivity was uncovered by the first X-ray structure of a fungal SAT domain, highlighted by a covalent hexanoyl thioester intermediate in the SAT active site. The crystal structure of SAT domain will enable protein engineering efforts aimed at mixing and matching different IPKS modules for the biosynthesis of new compounds.


Substance abuse and criminality.
Substance abuse issues and the law have become intricately linked over the years. This article reviews the current research underlying the association between substance abuse and crime, and provides an overview of the pertinent issues in conducting a substance abuse evaluation in the forensic context. The epidemiology of substance abuse and crime is reviewed, exploring the association between crime and specific psychoactive substances. Clinical considerations underlying the association are discussed, with specific attention paid to the pharmacological effects of psychoactive substances and to the role of substance use in individuals with serious associated psychopathology. Diagnostic and etiological issues that are important in differentiating substance abuse from criminality are considered in the context of conducting forensic evaluations. Finally, key components of a forensic substance abuse evaluation are presented.


BREX system of Escherichia coli distinguishes self from non-self by methylation of a specific DNA site.
Prokaryotes evolved numerous systems that defend against predation by bacteriophages. In addition to well-known restriction-modification and CRISPR-Cas immunity systems, many poorly characterized systems exist. One class of such systems, named BREX, consists of a putative phosphatase, a methyltransferase and four other proteins. A Bacillus cereus BREX system provides resistance to several unrelated phages and leads to modification of specific motif in host DNA. Here, we study the action of BREX system from a natural Escherichia coli isolate. We show that while it makes cells resistant to phage λ infection, induction of λ prophage from cells carrying BREX leads to production of viruses that overcome the defense. The induced phage DNA contains a methylated adenine residue in a specific motif. The same modification is found in the genome of BREX-carrying cells. The results establish, for the first time, that immunity to BREX system defense is provided by an epigenetic modification.


The hypertriglyceridemic-waist phenotype is associated with the Framingham risk score and subclinical atherosclerosis in Canadian Cree.
For primary prevention of cardiovascular disease (CVD), Canadian guidelines recommend that asymptomatic Canadians with abdominal obesity undergo Framingham risk score (FRS) assessment, and that in Indigenous Peoples, indicators of metabolic syndrome also be used to identify at-risk individuals. The hypertriglyceridemic-waist phenotype (HTGW) has been proposed to be a surrogate marker of visceral obesity and a simple proxy measure for metabolic syndrome. The primary aim of this study was to evaluate whether the HTGW and the FRS associated with sub-clinical atherosclerosis. Asymptomatic Cree participants in a cross-sectional study conducted 2005-2009 (n = 446, 18-81 y) were assessed for the HTGW using NCEP-ATP-III gender-specific-cutoffs (waist circumference: for men, ≥102 cm; for women ≥88 cm) and fasting triglycerides ≥1.7 mmol/L. Sub-clinical atherosclerosis was defined by the presence of a high sex-specific common-carotid-intimal-medial-wall-thickness (≥75th percentile). HTGW was present in 26.7% and a 10-y FRS greater than 10% was present in 18.8% of participants. The multivariate adjusted OR (95% CI) for sub-clinical atherosclerosis associated with an FRS greater than 10% was 4.10 (2.20-7.50) while that associated with the HTGW phenotype was 1.74 (95% CI 1.61-1.88) from a model including age, body mass index, alcohol consumption, FRS and the HTGW. The HTGW phenotype is prevalent in the Cree. Our findings support further study on the utility of combining the HTGW with the FRS in the prediction of cardiovascular disease outcomes and in health screening and intervention programs among indigenous peoples.


Nuclear receptor LRH-1 induces the reproductive neuropeptide kisspeptin in the hypothalamus.
The differential expression and secretion of the neuropeptide kisspeptin from neurons in the arcuate (Arc) and anteroventral periventricular (AVPV) nuclei of the hypothalamus coordinate the temporal release of pituitary gonadotropins that control the female reproductive cycle. However, the molecular basis for this differential regulation is incompletely understood. Here, we report that liver receptor homolog-1 (LRH-1), a member of the nuclear receptor superfamily, is expressed in kisspeptin neurons in the Arc but not in the AVPV in female mice. LRH-1 binds directly to the kisspeptin (Kiss1) promoter and stimulates Kiss1 transcription. Deletion of LRH-1 from kisspeptin neurons in mice decreased Kiss1 expression in the Arc, leading to reduced plasma FSH levels, dysregulated follicle maturation, and prolongation of the estrous cycle. Conversely, overexpression of LRH-1 in kisspeptin neurons increased Arc Kiss1 expression and plasma FSH concentrations. These studies provide a molecular basis for the differential regulation of basal kisspeptin expression in Arc and AVPV neurons and reveal a prominent role for LRH-1 in hypothalamus in regulating the female reproductive axis.


Proposal for tunable dual channel transmitter and mechano-optical switch based on photonic crystal.
In this article we propose a novel mechano-optical switch and dual channel transmitter based on photonic crystal. The device consists of two waveguides and an elliptical cavity in a square lattice structure. Two optical signals at separate wavelengths are inserted in the input waveguide. The elliptical cavity can be rotated using a mechanical force, which results in the control of transmission efficiency at each of the wavelengths. In addition, rotation of the cavity can be considered as a switching action, which changes on-off states of the output signals.


Polycythaemia in infants of diabetic mothers: β-hydroxybutyrate stimulates erythropoietic activity.
This study tested whether elevated maternal β-hydroxybutyrate (β-OHB) levels contribute to polycythaemia in infants of diabetic mothers. Pregnant diabetic women (n = 27) and non-diabetic controls (n = 20) and their singleton infants were included. Maternal glycosylated haemoglobin and β-OHB levels were studied at 34-36 weeks' gestation; levels were significantly higher in mothers with diabetes than in controls. Birth weights and cord blood levels of insulin and fetal haemoglobin were significantly higher in infants from diabetic mothers compared with control infants, as were haematocrit levels in venous blood samples taken from each infant at 4 h following delivery. Cord blood erythropoietin levels were similar in both groups. There was a positive strong correlation between maternal β-OHB levels and polycythaemia in newborn infants, indicating that β-OHB could activate erythropoiesis independently from intrauterine hyperinsulinaemia and/or erythropoietin levels, and may be important in the pathogenesis of polycythaemia in infants born to diabetic mothers.


Prospective analysis of circulating metabolites and breast cancer in EPIC.
Metabolomics is a promising molecular tool to identify novel etiologic pathways leading to cancer. Using a targeted approach, we prospectively investigated the associations between metabolite concentrations in plasma and breast cancer risk. A nested case-control study was established within the European Prospective Investigation into Cancer cohort, which included 1624 first primary incident invasive breast cancer cases (with known estrogen and progesterone receptor and HER2 status) and 1624 matched controls. Metabolites (n = 127, acylcarnitines, amino acids, biogenic amines, glycerophospholipids, hexose, sphingolipids) were measured by mass spectrometry in pre-diagnostic plasma samples and tested for associations with breast cancer incidence using multivariable conditional logistic regression. Among women not using hormones at baseline (n = 2248), and after control for multiple tests, concentrations of arginine (odds ratio [OR] per SD = 0.79, 95% confidence interval [CI] = 0.70-0.90), asparagine (OR = 0.83 (0.74-0.92)), and phosphatidylcholines (PCs) ae C36:3 (OR = 0.83 (0.76-0.90)), aa C36:3 (OR = 0.84 (0.77-0.93)), ae C34:2 (OR = 0.85 (0.78-0.94)), ae C36:2 (OR = 0.85 (0.78-0.88)), and ae C38:2 (OR = 0.84 (0.76-0.93)) were inversely associated with breast cancer risk, while the acylcarnitine C2 (OR = 1.23 (1.11-1.35)) was positively associated with disease risk. In the overall population, C2 (OR = 1.15 (1.06-1.24)) and PC ae C36:3 (OR = 0.88 (0.82-0.95)) were associated with risk of breast cancer, and these relationships did not differ by breast cancer subtype, age at diagnosis, fasting status, menopausal status, or adiposity. These findings point to potentially novel pathways and biomarkers of breast cancer development. Results warrant replication in other epidemiological studies.


Mapping malaria risk in Bangladesh using Bayesian geostatistical models.
Background malaria-control programs are increasingly dependent on accurate risk maps to effectively guide the allocation of interventions and resources. Advances in model-based geostatistics and geographical information systems (GIS) have enabled researchers to better understand factors affecting malaria transmission and thus, more accurately determine the limits of malaria transmission globally and nationally. Here, we construct Plasmodium falciparum risk maps for Bangladesh for 2007 at a scale enabling the malaria-control bodies to more accurately define the needs of the program. A comprehensive malaria-prevalence survey (N = 9,750 individuals; N = 354 communities) was carried out in 2007 across the regions of Bangladesh known to be endemic for malaria. Data were corrected to a standard age range of 2 to less than 10 years. Bayesian geostatistical logistic regression models with environmental covariates were used to predict P. falciparum prevalence for 2- to 10-year-old children (PfPR(2-10)) across the endemic areas of Bangladesh. The predictions were combined with gridded population data to estimate the number of individuals living in different endemicity classes. Across the endemic areas, the average PfPR(2-10) was 3.8%. Environmental variables selected for prediction were vegetation cover, minimum temperature, and elevation. Model validation statistics revealed that the final Bayesian geostatistical model had good predictive ability. Risk maps generated from the model showed a heterogeneous distribution of PfPR(2-10) ranging from 0.5% to 50%; 3.1 million people were estimated to be living in areas with a PfPR(2-10) greater than 1%. Contemporary GIS and model-based geostatistics can be used to interpolate malaria risk in Bangladesh. Importantly, malaria risk was found to be highly varied across the endemic regions, necessitating the targeting of resources to reduce the burden in these areas.


Dynamic culture yields engineered myocardium with near-adult functional output.
Engineered cardiac tissues hold promise for cell therapy and drug development, but exhibit inadequate function and maturity. In this study, we sought to significantly improve the function and maturation of rat and human engineered cardiac tissues. We developed dynamic, free-floating culture conditions for engineering "cardiobundles", 3-dimensional cylindrical tissues made from neonatal rat cardiomyocytes or human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) embedded in fibrin-based hydrogel. Compared to static culture, 2-week dynamic culture of neonatal rat cardiobundles significantly increased expression of sarcomeric proteins, cardiomyocyte size (∼2.1-fold), contractile force (∼3.5-fold), and conduction velocity of action potentials (∼1.4-fold). The average contractile force per cross-sectional area (59.7 mN/mm2) and conduction velocity (52.5 cm/s) matched or approached those of adult rat myocardium, respectively. The inferior function of statically cultured cardiobundles was rescued by transfer to dynamic conditions, which was accompanied by an increase in mTORC1 activity and decline in AMPK phosphorylation and was blocked by rapamycin. Furthermore, dynamic culture effects did not stimulate ERK1/2 pathway and were insensitive to blockers of mechanosensitive channels, suggesting increased nutrient availability rather than mechanical stimulation as the upstream activator of mTORC1. Direct comparison with phenylephrine treatment confirmed that dynamic culture promoted physiological cardiomyocyte growth rather than pathological hypertrophy. Optimized dynamic culture conditions also augmented function of human cardiobundles made reproducibly from cardiomyocytes derived from multiple hPSC lines, resulting in significantly increased contraction force (∼2.5-fold) and conduction velocity (∼1.4-fold). The average specific force of 23.2 mN/mm2 and conduction velocity of 25.8 cm/s approached the functional metrics of adult human myocardium. In conclusion, we have developed a versatile methodology for engineering cardiac tissues with a near-adult functional output without the need for exogenous electrical or mechanical stimulation, and have identified mTOR signaling as an important mechanism for advancing tissue maturation and function in vitro.


Transcript analysis and comparative evaluation of shaker and slowmo gene homologues from the European corn borer, Ostrinia nubilalis.
The movement and dispersal of larval Lepidoptera impact their survival and distribution within the natural landscape. Homologues of the Drosophila behaviour-linked genes shaker (shkr) and slowmo (slmo) were identified from Ostrinia nubilalis (Lepidoptera: Crambidae). Onshkr was isolated as a 1610-nucleotide (nt) constitutively expressed transcript encoding a membrane-localized 469-amino-acid (aa) protein with a conserved tetramerization domain and the six-domain architecture necessary for the molecule to fold into an active K(+) channel. Three expressed splice variants of 682, 970 and 1604 nt were identified for the Onslmo gene, and encode predicted 141 and 228 aa proteins with a conserved protein of relevant evolutionary and lymphoid interest (PRELI) domain that may function in mitochondrial protein sorting and perinuclear protein localization. Onshkr and Onslmo protein sequences aligned within monophyletic lepidopteran groups.


Meeting recommended dietary intakes in meal plans with ≥4 servings of grain-based foods daily.
To develop meal plans using grain-based foods demonstrating how to incorporate wholegrain foods into a balanced diet for weight maintenance for different cuisines. The present study examines the ability of meal plans with ≥4 grain-based servings daily to meet nutrient recommendations using lacto-ovo vegetarian and rice-based cuisines. Eighteen plans from each cuisine for three age brackets for both genders were developed. Plans aimed for ≥4 servings of grain-based foods daily, with separate plans for all wholegrain, all refined-grain and half wholegrain-half refined-grain foods. Meal plans followed an isoenergetic approach and were designed to meet specific Australian nutrient reference values and serving sizes. All plans met the Recommended Dietary Intake or Adequate Intake for targeted nutrients except for Fe in the rice-based meal plan for females aged ≥19 years (17 mg). In the plans for 14-18 year and ≥19 year age groups, four servings of grain-based foods could be accommodated. In the plans for 9-13 years, increasing the number of grain-based food servings to four reduced micronutrients levels delivered by the total diet. Specific food choices were made to ensure nutrient targets were met across each category for wholegrain and refined-grain plans. The major difference in nutrients between wholegrain and refined-grain foods was found in the vegetarian cuisine, where the meal plans containing whole grains produced on average 30 % higher fibre (38-53 g) levels than those with refined grains (27-40 g). With careful food selection, meal plans with ≥4 servings of grain-based foods daily can meet nutrient reference values for lacto-ovo vegetarian and rice-based cuisines.


Microfluidic micropipette aspiration for measuring the deformability of single cells.
We present a microfluidic technique for measuring the deformability of single cells using the pressure required to deform such cells through micrometre-scale tapered constrictions. Our technique is equivalent to whole-cell micropipette aspiration, but involves considerably simpler operation, less specialized equipment, and less technical skill. Single cells are infused into a microfluidic channel, and then deformed through a series of funnel-shaped constrictions. The constriction openings are sized to create a temporary seal with each cell as it passes through the constriction, replicating the interaction with the orifice of a micropipette. Precisely controlled deformation pressures are generated using an external source and then attenuated 100 : 1 using an on-chip microfluidic circuit. Our apparatus is capable of generating precisely controlled pressures as small as 0.3 Pa in a closed microchannel network, which is impervious to evaporative losses that normally limit the precision of such equipment. Intrinsic cell deformability, expressed as cortical tension, is determined from the threshold deformation pressure using the liquid-drop model. We measured the deformability of several types of nucleated cells and determined the optimal range of constriction openings. The cortical tension of passive human neutrophils was measured to be 37.0 ± 4.8 pN μm(-1), which is consistent with previous micropipette aspiration studies. The cortical tensions of human lymphocytes, RT4 human bladder cancer cells, and L1210 mouse lymphoma cells were measured to be 74.7 ± 9.8, 185.4 ± 25.3, and 235.4 ± 31.0 pN μm(-1) respectively. The precision and usability of our technique demonstrates its potential as a biomechanical assay for wide-spread use in biological and clinical laboratories.


Propionate stimulates pyruvate oxidation in the presence of acetate.
Flux through pyruvate dehydrogenase (PDH) in the heart may be reduced by various forms of injury to the myocardium, or by oxidation of alternative substrates in normal heart tissue. It is important to distinguish these two mechanisms because imaging of flux through PDH based on the appearance of hyperpolarized (HP) [(13)C]bicarbonate derived from HP [1-(13)C]pyruvate has been proposed as a method for identifying viable myocardium. The efficacy of propionate for increasing PDH flux in the setting of PDH inhibition by an alternative substrate was studied using isotopomer analysis paired with exams using HP [1-(13)C]pyruvate. Hearts from C57/bl6 mice were supplied with acetate (2 mM) and glucose (8.25 mM). (13)C NMR spectra were acquired in a cryogenically cooled probe at 14.1 Tesla. After addition of hyperpolarized [1-(13)C]pyruvate, (13)C NMR signals from lactate, alanine, malate, and aspartate were easily detected, in addition to small signals from bicarbonate and CO2. The addition of propionate (2 mM) increased appearance of HP [(13)C]bicarbonate >30-fold without change in O2 consumption. Isotopomer analysis of extracts from the freeze-clamped hearts indicated that acetate was the preferred substrate for energy production, glucose contribution to energy production was minimal, and anaplerosis was stimulated in the presence of propionate. Under conditions where production of acetyl-CoA is dominated by the availability of an alternative substrate, acetate, propionate markedly stimulated PDH flux as detected by the appearance of hyperpolarized [(13)C]bicarbonate from metabolism of hyperpolarized [1-(13)C]pyruvate.


Responses of innate immune cells to group A Streptococcus.
Group A Streptococcus (GAS), also called Streptococcus pyogenes, is a Gram-positive beta-hemolytic human pathogen which causes a wide range of mostly self-limiting but also several life-threatening diseases. Innate immune responses are fundamental for defense against GAS, yet their activation by pattern recognition receptors (PRRs) and GAS-derived pathogen-associated molecular patterns (PAMPs) is incompletely understood. In recent years, the use of animal models together with the powerful tools of human molecular genetics began shedding light onto the molecular mechanisms of innate immune defense against GAS. The signaling adaptor MyD88 was found to play a key role in launching the immune response against GAS in both humans and mice, suggesting that PRRs of the Toll-like receptor (TLR) family are involved in sensing this pathogen. The specific TLRs and their ligands have yet to be identified. Following GAS recognition, induction of cytokines such as TNF and type I interferons (IFNs), leukocyte recruitment, phagocytosis, and the formation of neutrophil extracellular traps (NETs) have been recognized as key events in host defense. A comprehensive knowledge of these mechanisms is needed in order to understand their frequent failure against GAS immune evasion strategies.


Are perceptuo-motor decisions really more optimal than cognitive decisions?
Human high-level cognitive decisions appear sub-optimal (Kahneman, Slovic, & Tversky, 1982; Kahneman & Tversky, 1979). Paradoxically, perceptuo-motor decisions appear optimal, or nearly optimal (Trommershäuser, Maloney, & Landy, 2008). Here, we highlight limitations to the comparison of performance between and within domains. These limitations are illustrated by means of two perceptuo-motor decision-making experiments. The results indicate that participants did not optimize fundamental performance-related factors (precision and time usage), even though standard analyses may have classed participants as 'optimal'. Moreover, simulations and comparisons across our studies demonstrate that optimality depends on task difficulty. Thus, it seems that a standard model of perceptuo-motor decision-making fails to provide an absolute standard of performance. Importantly, this appears to be a limitation of optimal models of human behaviour in general. This, in conjunction with non-trivial evaluative- and methodological differences, suggests that verdicts favouring perceptuo-motor, or perceptual, systems over higher-level cognitive systems in terms of level of performance are premature.


Work-related asthma from cleaning agents versus other agents.
Cleaning agents have been commonly implicated as causative or triggering factors in work-related asthma (WRA), mainly from epidemiologic studies. Relatively few clinical series have been reported. We aimed to compare socio-demographic and clinical features among tertiary clinic patients with WRA exposed to cleaning and non-cleaning products. Analyses were conducted on a patient database containing 208 patients with probable WRA referred to the asthma and airway centre at a tertiary centre hospital in Canada from 2000 to 2014. Chi-squared and independent samples t-tests were used to analyse categorical and continuous data, respectively. Twenty-two (11%) WRA cases were attributed to a variety of cleaning product exposures, 12 were diagnosed as occupational asthma (OA) and 10 as work-exacerbated asthma (WEA) (10% of all OA and 11% of all WEA). There were multiple exposures and the responsible agent(s) could seldom be clearly identified. Most frequent categories of exposure were surfactants, alcohols, disinfectants and acids. Compared to WRA with other exposures, those with cleaning agent exposures had a significantly larger proportion of females (82 versus 35%, P < 0.001), included a higher percentage of workers in healthcare (41 versus 4%, P < 0.001), and submitted more workers' compensation claims (86 versus 64%, P = 0.05). Other characteristics were comparable. In a tertiary referral clinic, patients with WRA from cleaning agent exposure had clinical characteristics that were similar to those with WRA from other causes. Most frequent exposures were surfactants, alcohols, disinfectants and acids.


Transcatheter valve-in-valve implantation with the Edwards SAPIEN in patients with bioprosthetic heart valve failure: the Milan experience.
Reoperation for bioprosthetic heart valve failure is associated with significant morbidity and mortality, particularly in high-risk patients. Transcatheter valve-in-valve (VIV) implantation may offer a less invasive alternative. The aim of this study was to report our initial experience with transcatheter VIV implantation to treat degenerated tissue valves. VIV implantation with the Edwards SAPIEN transcatheter heart valve (THV; Edwards Lifesciences Inc, Irvine, CA, USA) was performed in 18 high-risk patients (STS 8.2±5.2%; logistic EuroSCORE 37.4±20.8%) with symptomatic bioprosthetic failure (17 aortic, one mitral). Valve Academic Research Consortium (VARC) definitions were applied for endpoint adjudication. Transfemoral access was the preferred vascular approach (16 patients, with the mitral VIV delivered anterogradely through the femoral vein; one transaxillary and one transapical). The majority (83%) of procedures were performed under local anaesthesia and sedation. Device success was achieved in all but one patient who had a final transaortic gradient ≥20mmHg. Acute kidney injury occurred in three patients (Stage 3 in 1), life-threatening or major bleeding in four patients, while major vascular complications occurred in one patient. Permanent pacemaker implantation was required in two patients. There were no deaths or neurological events at 30-day follow-up. At a median follow-up of 11 months (interquartile range 6-16), the mortality rate was 5.6% and all patients were in NYHA class II or lower. Transcatheter implantation of the Edwards THV within a degenerated aortic bioprosthesis, performed predominantly via the transfemoral route, is feasible and associated with good periprocedural and clinical outcomes in high-risk surgical patients.


The SUPERFAMILY database in 2004: additions and improvements.
The SUPERFAMILY database provides structural assignments to protein sequences and a framework for analysis of the results. At the core of the database is a library of profile Hidden Markov Models that represent all proteins of known structure. The library is based on the SCOP classification of proteins: each model corresponds to a SCOP domain and aims to represent an entire superfamily. We have applied the library to predicted proteins from all completely sequenced genomes (currently 154), the Swiss-Prot and TrEMBL databases and other sequence collections. Close to 60% of all proteins have at least one match, and one half of all residues are covered by assignments. All models and full results are available for download and online browsing at http://supfam.org. Users can study the distribution of their superfamily of interest across all completely sequenced genomes, investigate with which other superfamilies it combines and retrieve proteins in which it occurs. Alternatively, concentrating on a particular genome as a whole, it is possible first, to find out its superfamily composition, and secondly, to compare it with that of other genomes to detect superfamilies that are over- or under-represented. In addition, the webserver provides the following standard services: sequence search; keyword search for genomes, superfamilies and sequence identifiers; and multiple alignment of genomic, PDB and custom sequences.


Sensitivity of fatty acid desaturation and elongation to plasma zinc concentration: a randomised controlled trial in Beninese children.
Zn status may affect fatty acid (FA) metabolism because it acts as a cofactor in FA desaturase and elongase enzymes. Zn supplementation affects the FA desaturases of Zn-deficient rats, but whether this occurs in humans is unclear. We evaluated the associations between baseline plasma Zn (PZn) concentration and plasma total phospholipid FA composition, as well as the effect of daily consumption of Zn-fortified water on FA status in Beninese children. A 20-week, double-blind randomised controlled trial was conducted in 186 school age children. The children were randomly assigned to receive a daily portion of Zn-fortified, filtered water delivering on average 2·8 mg Zn/d or non-fortified filtered water. Plasma total phospholipid FA composition was determined using capillary GLC and PZn concentrations by atomic absorption spectrometry. At baseline, PZn correlated positively with dihomo-γ-linolenic acid (DGLA, r 0·182; P=0·024) and the DGLA:linoleic acid (LA) ratio (r 0·293; P<0·000), and negatively with LA (r -0·211; P=0·009) and the arachidonic acid:DGLA ratio (r -0·170; P=0·036). With the intervention, Zn fortification increased nervonic acid (B: 0·109; 95 % CI 0·001, 0·218) in all children (n 186) and more so in children who were Zn-deficient (n 60) at baseline (B: 0·230; 95 % CI 0·023, 0·488). In conclusion, in this study, Zn-fortified filtered water prevented the reduction of nervonic acid composition in the plasma total phospholipids of children, and this effect was stronger in Zn-deficient children. Thus, Zn status may play an important role in FA desaturation and/or elongation.


Evaluation of the injection pain with the use of DentalVibe injection system during supraperiosteal anaesthesia in children: a randomised clinical trial.
The purpose of this study was to compare the use of a traditional syringe (TS) and the DentalVibe (DV) Injection Comfort System on the pain of needle insertion and injection of supraperiosteal (SP) anaesthesia into the mandibles and maxillas of children aged 6-12 years. The study was a randomised, controlled, crossover clinical trial, comprising 60 children requiring an operative procedure with SP anaesthesia on both their mandibular and maxillary molars, bilaterally. One of the molars was treated with a TS, and the contralateral tooth was treated with the DV for both arches. On each visit, subjective and objective pain was evaluated using the Wong-Baker FACES Pain Rating Scale and the Face, Leg, Activity, Cry, Consolability Scale. Patients were asked which technique they preferred. The data were analysed using Wilcoxon signed-rank test, Spearman's correlation test, and Mann-Whitney U-test. No statistically significant differences were noted between TS and DV for pain during injection and needle insertion for supraperiosteal anaesthesia in either the maxillary and mandibular operative procedures. Children experienced similar pain during SP anaesthesia administered with a TS and the DV, regardless of gender and jaw differences. DV was less preferred over the traditional procedure in children.


Numerical comparison between conventional dispersion compensating fibers and photonic crystal fibers as lumped Raman amplifiers.
In this paper we discuss the use of photonic crystal fibers (PCFs) as discrete devices for simultaneous wideband dispersion compensation and Raman amplification. The performance of the PCFs in terms of gain, ripple, optical signal-to-noise ratio (OSNR) and required fiber length for complete dispersion compensation is compared with conventional dispersion compensating fibers (DCFs). The main goal is to determine the minimum PCF loss beyond which its performance surpasses a state-of-the-art DCF and justifies practical use in telecommunication systems.


Smoking-gender interaction and risk for rheumatoid arthritis.
The present case-control study was conducted to investigate the relationship between smoking and rheumatoid arthritis, and to investigate formally the interaction between sex, smoking, and risk for developing rheumatoid arthritis. The study was performed in the Central District of Finland. Cases were patients with rheumatoid arthritis and the control group was a random sample of the general population. Logistic regression models were used to evaluate the effect of smoking on risk for rheumatoid arthritis, after adjusting for the effects of age, education, body mass index, and indices of general health and pain. Overall, 1095 patients with rheumatoid arthritis and 1530 control individuals were included. Patients were older, less well educated, more disabled, and had poorer levels of general health as compared with control individuals (all P < 0.01). Preliminary analyses revealed the presence of substantial statistical interaction between smoking and sex (P < 0.001). In separate multivariable analyses, past history of smoking was associated with increased risk for rheumatoid arthritis overall in men (odds ratio 2.0, 95% confidence interval 1.2-3.2) but not in women. Among men, this effect was seen only for rheumatoid factor-positive rheumatoid arthritis. There were significant interactions between smoking and age among women but not among men. We conclude that sex is a biologic effect modifier in the association between smoking and rheumatoid arthritis. The role of menopause in the etiology of rheumatoid arthritis merits further research.


Population Genetic Analysis Reveals Diversity in Lasiodiplodia Species Infecting Date Palm, Citrus, and Mango in Oman and the UAE.
Lasiodiplodia is a common pathogen causing dieback, gummosis, or root necrosis on the three most important fruit crops in Oman and the United Arab Emirates (UAE): date palm (Phoenix dactylifera), Citrus spp., and mango (Mangifera indica). A study was conducted to examine diversity in 64 Lasiodiplodia isolates infecting date palm (24), Citrus (11), and mango (29) in Oman and the UAE. Identification based on sequences of the internal transcribed spacer (ITS) rDNA and EF1α gene showed that date palm isolates belonged to L. hormozganensis (75% of isolates) and L. theobromae (25%); Citrus isolates belonged to L. hormozganensis (45%), L. theobromae (45%), and L. iraniensis (10%); and mango isolates belonged to L. theobromae (59%), L. iraniensis (34%), and L. hormozganensis (7%). Amplified fragment length polymorphism (AFLP) fingerprinting of the 64 isolates using four primer pair combinations produced 64 genotypes and 972 polymorphic alleles. Cluster analysis separated the isolates into four clusters representing the three species. A higher level of genetic diversity was observed in L. iraniensis (0.3105) compared to L. hormozganensis (0.2503) and L. theobromae (0.2331) in Oman. Analysis of molecular variance (AMOVA) indicated the existence of low levels of genetic differentiation among date palm populations of L. hormozganensis obtained from Oman and the UAE (FST = 0.025) and among populations of L. hormozganensis (0.0485) and L. theobromae (0.0703) from date palm, Citrus, and mango. These findings imply a high rate of movement of L. hormozganensis and L. theobromae isolates among date palm, Citrus, and mango and between the two countries. Findings from the pathogenicity test supported the AMOVA analysis and suggested a lack of host specialization in L. hormozganensis, L. iraniensis, and L. theobromae on date palm, acid lime, and mango. Although this is the first record of L. hormozganensis and L. iraniensis in Oman, the relatively moderate level of genetic diversity in the two species compared to L. theobromae suggests that the two species have been in Oman for a long time but misidentified by morphology and ITS rDNA sequences as L. theobromae. This study is also the first record of date palm and acid lime as natural hosts for L. hormozganensis and the first record of L. hormozganensis in the UAE. The diversity in Lasiodiplodia species affecting date palm, Citrus, and mango in Oman and the UAE should be taken into consideration when planning future management programs for diseases caused by these pathogens.


Photodynamic UVA-riboflavin bacterial elimination in antibiotic-resistant bacteria.
To evaluate the bactericidal effect of clinical ultraviolet A (UVA) settings used in photoactivated chromophore for infectious keratitis (PACK)-collagen cross-linking (CXL) in antibiotic-resistant and non-resistant bacterial strains. Well-characterized bacterial strains from clinical isolates, without and with antibiotic resistance, were studied in a pairwise comparison. The evaluated pathogens were Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, and Enterococcus faecalis. Bacteria were dispersed in PBS and diluted to a concentration of approximately 4 × 105 /ml. Riboflavin was added to a concentration of 0.01%. By spreading the solution on a microscope slide, a fluid film layer, with a thickness of around 400 mm, was formed and UVA exposure followed. Eight separate exposures were made for each strain (n = 8). The degree of elimination in resistant and non-resistant pathogens was compared. The bactericidal efficacy of exposure differed between the tested microorganisms, and the mean elimination ranged between 60 and 92%, being most extensive in both of the evaluated Pseudomonas strains and least in the E. faecalis strains. Similar reductions were seen in antibiotic-resistant and non-resistant strains, with the exception of S. aureus, in which the resistant strain metchicillin-resistant Staphylococcus aureus (MRSA) was eradicated in a greater extent than the non-resistant strain (P = 0.030). UVA-riboflavin settings used in PACK-CXL are effective in reducing both antibiotic-resistant and non-resistant bacteria. Antibiotic resistance does not appear to be protective against the photooxidative exposure.


Generation and characterisation of functional recombinant antibody fragments against RNA replicase NIb from plum pox virus.
A monoclonal antibody (mAb 2A) able to react against the RNA replicase NIb from plum pox virus (PPV) was obtained and used for generating a specific scFv fragment. The VH and VL coding sequences were cloned and expressed as a fusion scFv protein to alkaline phosphatase. This fusion protein was able to recognise viral NIb in both Western and tissue-print ELISA blots. The affinity and specificity of scFv2A for NIb was similar to that of the parental mAb and the region YLEAFY from PPV-NIb was identified by PEPSCAN assay as the putative epitope. Isolated VH domains from scFv2A were also expressed as fusion to alkaline phosphatase. However, their ability to react against NIb was greatly altered. scFv2A fragments were transiently expressed in the cytosol of Nicotiana benthamiana and although they accumulated to low levels, inhibition-ELISA results indicated that they retained antigen-binding activity.


Interpreting clinical evidence of malingering: a Bayesian perspective.
Customary ways of reporting on or testifying about malingering have shortcomings. Stating an opinion "with reasonable medical certainty" tells fact-finders little about how much confidence the opinion deserves; stating that an individual's behavior is similar to that of known malingerers does not convey the information that fact-finders really need to know, which is the likelihood that the evaluee in question is a malingerer, given the evaluator's findings. Mossman and Hart (Mossman D, Hart KJ: Presenting evidence of malingering to courts: insights from decision theory. Behav. Sci. Law 14:271-91, 1996) recommend that mental health professionals address this problem by using Bayes' theorem to interpret test data from evaluations. However, these authors do not discuss the use of evidence obtained during interviews and from other clinical contexts, nor do they describe a method for quantifying imprecision in Bayesian probabilities. This article provides examples of how forensic evaluators might use a Bayesian perspective to interpret clinical indicia of malingering observed during evaluations of adjudicatory competence. The article discusses sources of imprecision in Bayesian posterior probabilities, describes a method for characterizing that imprecision using confidence intervals, and then presents several sample calculations that illustrate how interview findings change the likelihood of malingering. The article also discusses the implications of the Bayesian approach for forensic evaluations and for future research on malingered incompetence.


Natural control of HIV infection in young women in South Africa: HPTN 068.
Some individuals control HIV replication without antiretroviral (ARV) therapy. To analyze viral suppression in young women in rural South Africa enrolled in a trial evaluating a behavioral intervention for HIV prevention. Plasma samples were obtained from women ages 13-24 (81 infected at enrollment, 164 seroconverters). ARV testing was performed using an assay that detects 20 ARV drugs. Women were classified as viremic controllers if they were virally suppressed for ≥12 months with no ARV drug use. Samples from 216/245 (88.2%) women had no ARV drugs detected at their first HIV-positive visit. Thirty-four (15.7%) of the 216 women had a viral load <2,000 copies/mL. Fifteen of the 34 women were followed for ≥12 months; 12 were virally suppressed with no ARV drugs detected during follow-up. These women were classified as viremic controllers (overall: 12/216 = 5.6%). The median CD4 cell count at the first HIV-positive visit was higher among the 12 controllers than among the 204 women who were not using ARV drugs (759 vs. 549 cells/mm3, p = 0.02). Some women had a viral load <40 copies/mL at a single study visit, but none were classified as elite controllers (viral load <40 copies/mL for ≥12 months with no ARV drug use). In this cohort, 5.6% of women who were not using ARV drugs had sustained viral suppression. This represents a minimum estimate of the frequency of viremic controllers in this cohort, since some women were not followed long enough to meet the criteria for classification.


Activation of Aurora-A kinase by protein partner binding and phosphorylation are independent and synergistic.
Protein kinases are activated by phosphorylation and by the binding of activator proteins. The interplay of these two factors is incompletely understood. We applied energetic analysis to this question and characterized the activation process of the serine/threonine kinase Aurora-A by phosphorylation and by its protein partner, targeting protein for Xenopus kinesin-like protein 2 (TPX2). We discovered that these two activators act synergistically and without a predefined order: each can individually increase the activity of Aurora-A, and the effect of both bound together is the exact sum of their individual contributions to catalysis. Unexpectedly, the unphosphorylated enzyme has catalytic activity that is increased 15-fold by the binding of TPX2 alone. The energetic contribution of phosphorylation to catalysis is 2-fold greater than that of TPX2 binding, which is independent of the phosphorylation state of the enzyme. Based on this analysis, we propose a revised, fluid model of Aurora-A activation in which the first step is a reduction in the mobility of the activation loop by either TPX2 binding or phosphorylation. Furthermore, our results suggest that unphosphorylated Aurora-A bound to the mitotic spindle by TPX2 is catalytically active and that the phosphorylation state of Aurora-A is an inaccurate surrogate for its activity. Extending this form of analysis will allow us to compare quantitatively the effects of the whole network of kinase-activating partners. Comparison with other kinases showed that kinetic characterization detects those kinases whose activation loops undergo a rearrangement upon phosphorylation and thus whose unphosphorylated state offers a distinct target for the development of Type II inhibitors.


Demographics of the gay and lesbian population in the United States: evidence from available systematic data sources.
This work provides an overview of standard social science data sources that now allow some systematic study of the gay and lesbian population in the United States. For each data source, we consider how sexual orientation can be defined, and we note the potential sample sizes. We give special attention to the important problem of measurement error, especially the extent to which individuals recorded as gay and lesbian are indeed recorded correctly. Our concern is that because gays and lesbians constitute a relatively small fraction of the population, modest measurement problems could lead to serious errors in inference. In examining gays and lesbians in multiple data sets we also achieve a second objective: We provide a set of statistics about this population that is relevant to several current policy debates.


Measuring Protein Content in Food: An Overview of Methods.
In order to determine the quantity of protein in food, it is important to have standardized analytical methods. Several methods exist that are used in different food industries to quantify protein content, including the Kjeldahl, Lowry, Bradford and total amino acid content methods. The correct determination of the protein content of foods is important as, often, as is the case with milk, it determines the economic value of the food product and it can impact the economic feasibility of new industries for alternative protein production. This editorial provides an overview of different protein determination methods and describes their advantages and disadvantages.