Simple Culture Methods and Treatment to Study Hormonal Regulation of Ovule Development.
Ovule development is one of the most important processes in the reproductive development of higher plants and is a determinant of seed quality and quantity. Phytohormones play key roles in this process since loss-of-function mutants in hormone signaling show defective ovule phenotypes and reduced fertility. However, it is difficult to distinguish the direct effects of hormones on ovule development because it is parts of reproductive development and the defective phenotypes would be the indirect effects following the defective vegetative development. The treatment of hormones is a direct method to investigate the hormonal regulation of ovule development, but ovule is embedded inside several layers of floral organs, and traditional methods for hormone (or inhibitor) treatments have various limitations. We have developed simple methods to apply treatments to the flowers in a living plant, where an inflorescence apex is immersed into a solution in an inverted tube. We have also developed a specific system to culture and treat excised flowers/pistils. These procedures will be useful for research on the hormonal regulation of ovule development. We provide examples of how treatments with brassinosteroids (BR) and BR biosynthesis inhibitor. We cultured and treated plant materials using our newly developed methods, and observed the morphology of wild type ovules and fluorescence signals in a marker line to monitor the progress of ovule development. The results demonstrate BR promotes ovule development and our new methods are efficient and repeatable.


Affection of the Respiratory Muscles in Combined Complex I and IV Deficiency.
Combined complex I+IV deficiency has rarely been reported to manifest with the involvement of the respiratory muscles. A 45y male was admitted for hypercapnia due to muscular respiratory insufficiency. He required intubation and mechanical ventilation. He had a previous history of ophthalmoparesis since age 6y, ptosis since age 23y, and anterocollis since at least age 40y. Muscle biopsy from the right deltoid muscle at age 41y was indicative of mitochondrial myopathy. Biochemical investigations revealed a combined complex I+IV defect. Respiratory insufficiency was attributed to mitochondrial myopathy affecting not only the extra-ocular and the axial muscles but also the shoulder girdle and respiratory muscles. In addition to myopathy, he had mitochondrial neuropathy, abnormal EEG, and elevated CSF-protein. Possibly, this is why a single cycle of immunoglobulins was somehow beneficial. For muscular respiratory insufficiency he required tracheostomy and was scheduled for long-term intermittent positive pressure ventilation. Mitochondrial myopathy due to a combined complex I+IV defect with predominant affection of the extra-ocular muscles may progress to involvement of the limb-girdle, axial and respiratory muscles resulting in muscular respiratory insufficiency. In patients with mitochondrial myopathy, neuropathy and elevated cerebrospinal fluid protein, immunoglobulins may be beneficial even for respiratory functions.


A Proposal for Determining a Diagnostic Cutoff Value of a New Ultrasonographic Index of Retronychia Inversely Correlating with Disease Severity.
Ultrasonography has demonstrated a shortening of the distance between the origin of the nail plate and the base of the distal phalanx in retronychia. The aim of this study was to analyze the clinical and ultrasonographic features of retronychia. We evaluated the clinical findings in 18 patients with retronychia, along with the ratio of ultrasonographic distance a, extending between the nail plate origin and the base of the distal phalanx, to distance b, perpendicular to distance a, extending between the nail plate origin and the upper margin of the distal phalanx. Retronychia was present in 26 nails. The mean ± SD distance a was 7.66 ± 1.64 mm and distance b was 3.56 ± 1.95 mm. The mean ± SD a/b ratio was 2.59 ± 1.11. There was a significant inverse correlation between a/b ratio and clinical severity (Pearson correlation = -0.668; P < .001). The cutoff value of this ratio was 3.319, with specificity of 90% and sensitivity of 69%. The ratio of distance a/distance b and the cutoff value of this ratio may help in making the diagnosis, in objectively determining the disease severity, and in selecting a patient-specific treatment approach.


Spectrum of CFTR mutations on Réunion Island: impact on neonatal screening.
The large heterogeneity in the cystic fibrosis (CF) gene is the main difficulty for genotype characterization. Numerous studies have reported considerable variations in frequencies of CF transmembrane conductance regulator (CFTR) mutations in different populations, such as African, Asian, or European populations. To completely characterize the spectrum of mutations in the CFTR gene in the Réunion Island population, we screened 228 CF chromosomes using denaturing high-pressure liquid chromatography and denaturing gradient gel electrophoresis following by direct sequencing. We identified 27 mutations, accounting for 93% of CF chromosomes. They included three novel mutations (M1T, 3121-3C-->G, and L1324P), which are described in this paper. The detection of such a high proportion of Réunion Island CFTR mutations is important for improving neonatal screening of CF on Réunion Island.


Antiproliferative effect of chitosan-added kimchi in HT-29 human colon carcinoma cells.
The anticancer effects of chitosan-added kimchi were investigated by using an in vitro cellular system with HT-29 human colon carcinoma cells. Two different kinds of chitosan-soluble chitosan with a 90% degree of deacetylation and 3 cps viscosity and nonsoluble chitosan with a 95% degree of deacetylation and 22 cps viscosity-were used as sub-ingredients to increase anticancer effects of kimchi. The soluble chitosan-added kimchi (SK) and nonsoluble chitosan-added kimchi (NK) were stronger growth inhibitors in HT-29 cells than the control kimchi (CK) according to the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and the growth inhibition test. Treatment with SK and NK induced apoptosis, as determined by 4,6-diamidino-2-phenylindole staining, and resulted in the up-regulation of Bax expression and down-regulation of Bcl-2, cIAP-1, cellular inhibitor of apoptosis-2, cyclooxygenase-2, inhibitory nitric oxide synthase, and nuclear factor kappaB (NF-kappaB) expressions when compared to CK. The antiproliferative and anti-apoptotic effects appeared to be more pronounced in the cells treated with NK. The antiproliferative effects of the chitosan-added kimchi appeared to be associated with the induction of apoptosis through NF-kappaB or an NF-kappaB-dependent pathway. These results suggest that chitosan has potential to be a valuable active ingredient in functional kimchi products with anticancer effects.


Comparative studies using the Morris water maze to assess spatial memory deficits in two transgenic mouse models of Alzheimer's disease.
Evaluation of the efficacy of novel therapeutics for potential treatment of Alzheimer's disease (AD) requires an animal model that develops age-related cognitive deficits reproducibly between independent groups of investigators. Herein we assessed comparative temporal changes in spatial memory function in two commercially available transgenic mouse models of AD using the Morris water maze (MWM), incorporating both visible and hidden platform training. Individual cohorts of cDNA-based 'line 85'-derived double-transgenic mice coexpressing the 'Swedish' mutation of amyloid precursor protein (APPSwe) and the presenillin 1 (PS1) 'dE9' mutation were assessed in the MWM at mean ages of 3.6, 9.3 and 14.8 months. We found significant deficits in spatial memory retention in APPSwe/PS1dE9 mice aged 3.6 months and robust deficits in spatial memory acquisition and retention in APPSwe/PS1dE9 mice aged 9.3 months, with a further significant decline by age 14.8 months. β-Amyloid deposits were present in brain sections by 7.25 months of age. In contrast, MWM studies with individual cohorts (aged 4-21 months) of single-transgenic genomic-based APPSwe mice expressing APPSwe on a yeast artificial chromosomal (YAC) construct showed no significant deficits in spatial memory acquisition until 21 months of age. There were no significant deficits in spatial memory retention up to 21 months of age and β-amyloid deposits were not present in brain sections up to 24 months of age. These data, generated using comprehensive study designs, show that APPSwe/PS1dE9 but not APPSwe YAC mice appear to provide a suitably robust model of AD for efficacy assessment of novel AD treatments in development.


Factors associated with mosquito net use by individuals in households owning nets in Ethiopia.
Ownership of insecticidal mosquito nets has dramatically increased in Ethiopia since 2006, but the proportion of persons with access to such nets who use them has declined. It is important to understand individual level net use factors in the context of the home to modify programmes so as to maximize net use. Generalized linear latent and mixed models (GLLAMM) were used to investigate net use using individual level data from people living in net-owning households from two surveys in Ethiopia: baseline 2006 included 12,678 individuals from 2,468 households and a sub-sample of the Malaria Indicator Survey (MIS) in 2007 included 14,663 individuals from 3,353 households. Individual factors (age, sex, pregnancy); net factors (condition, age, net density); household factors (number of rooms [2006] or sleeping spaces [2007], IRS, women's knowledge and school attendance [2007 only], wealth, altitude); and cluster level factors (rural or urban) were investigated in univariate and multi-variable models for each survey. In 2006, increased net use was associated with: age 25-49 years (adjusted (a) OR = 1.4, 95% confidence interval (CI) 1.2-1.7) compared to children U5; female gender (aOR = 1.4; 95% CI 1.2-1.5); fewer nets with holes (Ptrend = 0.002); and increasing net density (Ptrend < 0.001). Reduced net use was associated with: age 5-24 years (aOR = 0.2; 95% CI 0.2-0.3). In 2007, increased net use was associated with: female gender (aOR = 1.3; 95% CI 1.1-1.6); fewer nets with holes (aOR [all nets in HH good] = 1.6; 95% CI 1.2-2.1); increasing net density (Ptrend < 0.001); increased women's malaria knowledge (Ptrend < 0.001); and urban clusters (aOR = 2.5; 95% CI 1.5-4.1). Reduced net use was associated with: age 5-24 years (aOR = 0.3; 95% CI 0.2-0.4); number of sleeping spaces (aOR [per additional space] = 0.6, 95% CI 0.5-0.7); more old nets (aOR [all nets in HH older than 12 months] = 0.5; 95% CI 0.3-0.7); and increasing household altitude (Ptrend < 0.001). In both surveys, net use was more likely by women, if nets had fewer holes and were at higher net per person density within households. School-age children and young adults were much less likely to use a net. Increasing availability of nets within households (i.e. increasing net density), and improving net condition while focusing on education and promotion of net use, especially in school-age children and young adults in rural areas, are crucial areas for intervention to ensure maximum net use and consequent reduction of malaria transmission.


Impairment of humoral immunity to Plasmodium falciparum malaria in pregnancy by HIV infection.
HIV infection increases the risk of malaria infection in pregnant women. Antibodies to variant surface antigens (VSA) on infected erythrocytes might protect against malaria in pregnancy. We postulated that HIV-induced impairment of humoral immunity to VSA mediates the increased susceptibility to malaria. We compared serum concentrations of antibodies to VSA by flow cytometry or agglutination, and to merozoite proteins AMA-1 and MSP119 by ELISA, in 298 pregnant Malawian women, and related the findings to malaria and HIV infection, CD4-positive T-cell count, and HIV-1 viral load. Concentrations of IgG to placental type VSA were lower in HIV-infected women than in HIV-uninfected women (median 8 units [IQR 4-23] vs 20 [12-30]; p<0.0001), among women with malaria (p=0.009) and those without malaria (p=0.0062). The impairment was greatest in first pregnancy. Agglutinating antibodies to placental VSA were present in a lower proportion of HIV-infected than HIV-uninfected women (58 [35.1%] of 165 vs 50 [53.8%] of 93, p<0.001). The degree of antibody binding by flow cytometry was correlated with CD4-positive T-cell count (r=0.16, p=0.019) and inversely with HIV-1 viral load (r=-0.16, p=0.030). Concentrations of antibodies to AMA-1 were lower in HIV infection (p<0.0001) but were not correlated with CD4-positive T-cell count or viral load. Responses to MSP119 were little affected by HIV infection. In multivariate analyses, HIV was negatively associated with amount of antibody to both VSA and AMA-1 (p<0.001 for each) but not MSP119. HIV infection impairs antimalarial immunity, especially responses to placental type VSA. The impairment is greatest in the most immunosuppressed women and could explain the increased susceptibility to malaria seen in pregnant women with HIV infection.


Gadd45b knockout mice exhibit selective deficits in hippocampus-dependent long-term memory.
Growth arrest and DNA damage-inducible β (Gadd45b) has been shown to be involved in DNA demethylation and may be important for cognitive processes. Gadd45b is abnormally expressed in subjects with autism and psychosis, two disorders associated with cognitive deficits. Furthermore, several high-throughput screens have identified Gadd45b as a candidate plasticity-related gene. However, a direct demonstration of a link between Gadd45b and memory has not been established. The current studies first determined whether expression of the Gadd45 family of genes was affected by contextual fear conditioning. Gadd45b, and to a lesser extent Gadd45g, were up-regulated in the hippocampus following contextual fear conditioning, whereas Gadd45a was not. Next, Gadd45b knockout mice were tested for contextual and cued fear conditioning. Gadd45b knockout mice exhibited a significant deficit in long-term contextual fear conditioning; however, they displayed normal levels of short-term contextual fear conditioning. No differences between Gadd45b knockout and wild-type mice were observed in cued fear conditioning. Because cued fear conditioning is hippocampus independent, while contextual fear conditioning is hippocampus dependent, the current studies suggest that Gadd45b may be important for long-term hippocampus-dependent memory storage. Therefore, Gadd45b may be a novel therapeutic target for the cognitive deficits associated with many neurodevelopmental, neurological, and psychiatric disorders.


Mechanisms of block of muscle type CLC chloride channels (Review).
CLC proteins are a nine-member gene family of Cl- channels that have diverse roles in the plasma membrane and in intracellular organelles. The recent structure determination of bacterial CLC homologues by Dutzler et al. was a breakthrough for the structure-function analysis of CLC channels. This review describes the mechanisms of inhibition of muscle type CLC channels by two classes of small organic substances: 9-anthracene carboxylic acid (9AC) and p-chlorophenoxy propionic acid (CPP). Both substances block muscle type CLC channels (CLC-0 and CLC-1) from the intracellular side. For CPP, one could show that it inhibits the individual protopores of the double-barrelled channel. A major difference between the two types of blockers is the extremely slow binding- and unbinding-kinetics of 9AC (time scale of min), compared to that of CPP block (time scale of s), while the general mechanism of block seems to be quite similar. In the case of the chiral CPP only the S(-) enantiomer is effective. Both substances exhibit a strongly voltage-dependent block with strong inhibition at negative voltages and relief of block at depolarizing potentials at which the channels tend to open maximally. A quantitative kinetic model was developed for the CPP block of CLC-0 in which the closed state has a much larger affinity for CPP than the open state and opening of drug-bound channels is greatly slowed compared to drug-free channels. First experiments with mutated CLC-0 channels and with derivatives of CPP strongly support the pore localization of the CPP binding site. This work provides the basis for the use of these small organic substances as tools to investigate the pharmacological properties of mammalian CLC channels guided by the crystallographic structure of bacterial CLC homologues. They might also turn out to be useful to obtain information about the intricate coupling of gating and permeation that characterizes CLC channels.


Supraphysiological doses of levothyroxine alter regional cerebral metabolism and improve mood in bipolar depression.
Supplementation of standard treatment with high-dose levothyroxine (L-T(4)) is a novel approach for treatment-refractory bipolar disorders. This study tested for effects on brain function associated with mood alterations in bipolar depressed patients receiving high-dose L-T(4) treatment adjunctive to ongoing medication (antidepressants and mood stabilizers). Regional activity and whole-brain analyses were assessed with positron emission tomography and [(18)F]fluorodeoxyglucose in 10 euthyroid depressed women with bipolar disorder, before and after 7 weeks of open-label adjunctive treatment with supraphysiological doses of L-T(4) (mean dose 320 microg/day). Corresponding measurements were acquired in an age-matched comparison group of 10 healthy women without L-T(4) treatment. The primary biological measures were relative regional activity (with relative brain radioactivity taken as a surrogate index of glucose metabolism) in preselected brain regions and neuroendocrine markers of thyroid function. Treatment-associated changes in regional activity (relative to global activity) were tested against clinical response. Before L-T(4) treatment, the patients exhibited significantly higher activity in the right subgenual cingulate cortex, left thalamus, medial temporal lobe (right amygdala, right hippocampus), right ventral striatum, and cerebellar vermis; and had lower relative activity in the middle frontal gyri bilaterally. Significant behavioral and cerebral metabolic effects accompanied changes in thyroid hormone status. L-T(4) improved mood (remission in seven patients; partial response in three); and decreased relative activity in the right subgenual cingulate cortex, left thalamus, right amygdala, right hippocampus, right dorsal and ventral striatum, and cerebellar vermis. The decrease in relative activity of the left thalamus, left amygdala, left hippocampus, and left ventral striatum was significantly correlated with reduction in depression scores. Results of the whole-brain analyses were generally consistent with the volume of interest results. We conclude that bipolar depressed patients have abnormal function in prefrontal and limbic brain areas. L-T(4) may improve mood by affecting circuits involving these areas, which have been previously implicated in affective disorders.


Estimating the Population Benefits and Costs of Rituximab Therapy in the United States from 1998 to 2013 Using Real-World Data.
Rituximab was approved in 1997 and is regularly one of the largest drug expenditures for Medicare; however, its benefits and costs have not been estimated from a population perspective. To estimate both the clinical and the economic outcomes of rituximab for its approved hematological uses at the population level. Analyses using cancer registry incidence data from the Surveillance, Epidemiology, and End Results (SEER) program, and outcomes data from SEER data linked with Medicare administrative claims (SEER-Medicare data). These results were incorporated into an epidemiological simulation model of the population over time. We modeled all United States patients from 1998 to 2013 diagnosed with diffuse large B-cell lymphoma, follicular lymphoma, or chronic lymphocytic leukemia. Using this model, we estimated the life-years saved, as well as their economic benefit, in the United States population. We also estimated the incremental cost of adding rituximab to chemotherapy. All economic inputs were based on Medicare reimbursed amounts inflated to 2013 dollars. There were 279,704 cumulative life-years saved which were valued at $25.44 billion. The incremental direct medical cost of rituximab was estimated to be $8.92 billion, resulting in an incremental economic gain of $16.52 billion. These analyses, based on real-world evidence, show that the introduction of rituximab into clinical practice has produced a substantial number of incremental life-years. Importantly, the economic benefit of the life-years gained greatly exceeds the added costs of treatment.


The disabilities of the arm, shoulder and hand (DASH) outcome questionnaire: longitudinal construct validity and measuring self-rated health change after surgery.
The disabilities of the arm, shoulder and hand (DASH) questionnaire is a self-administered region-specific outcome instrument developed as a measure of self-rated upper-extremity disability and symptoms. The DASH consists mainly of a 30-item disability/symptom scale, scored 0 (no disability) to 100. The main purpose of this study was to assess the longitudinal construct validity of the DASH among patients undergoing surgery. The second purpose was to quantify self-rated treatment effectiveness after surgery. The longitudinal construct validity of the DASH was evaluated in 109 patients having surgical treatment for a variety of upper-extremity conditions, by assessing preoperative-to-postoperative (6-21 months) change in DASH score and calculating the effect size and standardized response mean. The magnitude of score change was also analyzed in relation to patients' responses to an item regarding self-perceived change in the status of the arm after surgery. Performance of the DASH as a measure of treatment effectiveness was assessed after surgery for subacromial impingement and carpal tunnel syndrome by calculating the effect size and standardized response mean. Among the 109 patients, the mean (SD) DASH score preoperatively was 35 (22) and postoperatively 24 (23) and the mean score change was 15 (13). The effect size was 0.7 and the standardized response mean 1.2.The mean change (95% confidence interval) in DASH score for the patients reporting the status of the arm as "much better" or "much worse" after surgery was 19 (15-23) and for those reporting it as "somewhat better" or "somewhat worse" was 10 (7-14) (p = 0.01). In measuring effectiveness of arthroscopic acromioplasty the effect size was 0.9 and standardized response mean 0.5; for carpal tunnel surgery the effect size was 0.7 and standardized response mean 1.0. The DASH can detect and differentiate small and large changes of disability over time after surgery in patients with upper-extremity musculoskeletal disorders. A 10-point difference in mean DASH score may be considered as a minimal important change. The DASH can show treatment effectiveness after surgery for subacromial impingement and carpal tunnel syndrome. The effect size and standardized response mean may yield substantially differing results.


Predictors of early acceptance of free spectacles provided to junior high school students in China.
To examine factors influencing adherence to spectacle wear and perceived value within a prospective 1-month trial of ready-made and custom spectacles in school-aged children with uncorrected refractive error in urban China. A total of 428 students aged 12 to 15 years with at least 1 diopter of uncorrected refractive error were given free spectacles and evaluated 1 month later at an unannounced visit. Demographic factors, vision, optical effects, and perceptions were modeled as predictors of observed use and perceived value using logistic regression adjusted for spectacle allocation. Of 415 students, 388 (93.5%) planned to use their spectacles, 227 (54.7%) valued their spectacles highly, 204 (49.2%) had their spectacles on hand, and 13 (3.0%) were lost to follow-up. Female students were 1.72 times (95% confidence interval [CI], 1.10-2.68), students from lower income households were 1.78 times (1.32-2.39), and those not concerned over appearance were 2.04 times (1.25-3.36) more likely to have spectacles on hand. Students with a pupil size of 4 mm or greater were 2.55 times (95% CI, 1.61-4.03) and students with spectacle vision worse than 20/20 were 2.06 times (1.20-3.49) more likely to have spectacles on hand. Self-report of high perceived value was 2.23 times (95% CI, 1.30-3.80) more likely with 20/20 spectacle vision, 1.63 times (1.06-2.52) more likely with base-in prismatic effects of 0.5 prism diopters or more, 3.52 times (2.03-6.13) more likely when students would not tolerate blur to avoid wearing spectacles, and 2.16 times (1.24-3.76) more likely with disbelief that spectacles would make vision worse. Spectacle type had no effect. Although most students planned to use their spectacles, only half were observed using them. Day-to-day use might increase if students were less concerned over appearance. Optical factors and beliefs surrounding spectacles are also predictive of acceptance. These findings provide further understanding of spectacle acceptance in teenagers.


Variation in performance times of elite flat-water canoeists from race to race.
To estimate variability in performance time and smallest worthwhile changes for elite flat-water canoeists competing in 200-, 500- or 1000-m events at international regattas. The data came from A and B finals held at 7 to 13 regattas in 2003 to 2007. A linear mixed-model analysis of log-transformed official race times provided estimates of variability as coefficients of variation and included terms to account for changes in performance between years, venues, and A and B finals. For men, the within-athlete variation in A finals was similar in canoeing and kayaking events, with the 200-m men's events demonstrating probably less variability than the longer events (by an overall factor of 0.75, x// 1.33) that may reflect differences in pacing strategies. In contrast, the within-athlete variation for women kayakers in A finals of the 500-m event was only half that of the other distances (ratio 0.54, x// 1.29), possibly because of differences in competitive experience or depth of competition. Predictability of performance in A finals was moderate to very high (interclass correlations 0.40 to 0.89). Within-athlete variation in the B finals was generally greater than in the A finals for the three distances for men, but there was no clear pattern for women. The smallest worthwhile changes in performance time (0.3 x within-athlete variability) in canoeing and kayaking are approx. 0.3% to 0.6%. Effects of 1% to 2% in power output would be required to achieve such changes in this generally highly predictable sport.


Potential health-related benefits of resistance training.
Public health guidelines primarily focus on the promotion of physical activity and steady-state aerobic exercise, which enhances cardiorespiratory fitness and has some impact on body composition. However, research demonstrates that resistance exercise training has profound effects on the musculoskeletal system, contributes to the maintenance of functional abilities, and prevents osteoporosis, sarcopenia, lower-back pain, and other disabilities. More recent seminal research demonstrates that resistance training may positively affect risk factors such as insulin resistance, resting metabolic rate, glucose metabolism, blood pressure, body fat, and gastrointestinal transit time, which are associated with diabetes, heart disease, and cancer. Research also indicates that virtually all the benefits of resistance training are likely to be obtained in two 15- to 20-min training sessions a week. Sensible resistance training involves precise controlled movements for each major muscle group and does not require the use of very heavy resistance. Along with brief prescriptive steady-state aerobic exercise, resistance training should be a central component of public health promotion programs.


Lipid peroxides and atherosclerosis.
Plasma lipid peroxide concentrations were measured in 100 patients with occlusive arterial disease proved angiographically (50 patients with ischaemic heart disease, 50 with peripheral arterial disease) and compared with values in 75 control patients with no clinical evidence of atherosclerosis. Lipid peroxide concentrations were significantly higher in patients both with ischaemic heart disease (median 4.37 mumol/l (interquartile range 3.85-5.75 mumol/l); p less than 0.001) and with peripheral arterial disease (median 4.37 mumol/l (3.88-5.21 mumol/l); p less than 0.001) than in controls (median 3.65 mumol/l (interquartile range 3.29-3.89 mumol/l). Overall there was a significant but weak correlation between plasma lipid peroxide and plasma triglyceride concentrations (rs = 0.25; p less than 0.001) but not between plasma lipid peroxide and plasma total cholesterol concentrations. Furthermore, hypertension, obesity, diabetes, smoking, positive family history, and intake of beta blockers and thiazide diuretics were not associated with significant differences in lipid peroxide values. This study provides clinical support to experimental data indicating that peroxidised lipids may be important in atherogenesis and its complications and also suggests that peroxidised lipids may provide an index of the severity of atherosclerosis.


Biatrial ablation vs. left atrial concomitant surgical ablation for treatment of atrial fibrillation: a meta-analysis.
Surgical ablation performed concomitantly with cardiac surgery has emerged as an effective curative strategy for atrial fibrillation (AF). Left atrial (LA) lesion sets for ablation have been suggested to reduce procedural times and post-surgical bradycardia compared with biatrial (BA) lesions. Given the inconclusive literature regarding BA vs. LA ablation, the present meta-analysis sought to assess the current evidence. Electronic searches were performed using six databases from their inception to December 2013, identifying all relevant randomized trials and observational studies comparing BA vs. LA surgical ablation AF patients undertaking cardiac surgery. In 10 included studies, 2225 patient results were available for analysis to compare BA (n = 888) vs. LA (n = 1337) ablation. Sinus rhythm prevalence was higher in the BA cohort compared with the LA cohort at 6-month and 12-month follow-up, but similar beyond 1 year. Permanent pacemaker implantations were higher in the BA cohort, but 30-day and late mortality, neurological events, and reoperation for bleeding were similar between BA and LA groups. Biatrial and LA ablations produced comparable 30-day and late mortality but LA was associated with significantly reduced permanent pacemaker implantation rates. Biatrial ablation appeared to be more efficacious than LA ablation in achieving SR at 1 year, but this difference was not maintained beyond 1 year. Trends appear to be driven by the preferential selection of long-standing and persistent AF patients for the BA approach. Future randomized studies of adequate follow-up are required to validate risks and benefits of BA vs. LA surgical ablation.


Circulating vitamin D, vitamin D-related genetic variation, and risk of fatal prostate cancer in the National Cancer Institute Breast and Prostate Cancer Cohort Consortium.
Evidence from experimental animal and cell line studies supports a beneficial role for vitamin D in prostate cancer (PCa). Although the results from human studies have been mainly null for overall PCa risk, there may be a benefit for survival. This study assessed the associations of circulating 25-hydroxyvitamin D (25(OH)D) and common variations in key vitamin D-related genes with fatal PCa. In a large cohort consortium, 518 fatal cases and 2986 controls with 25(OH)D data were identified. Genotyping information for 91 single-nucleotide polymorphisms (SNPs) in 7 vitamin D-related genes (vitamin D receptor, group-specific component, cytochrome P450 27A1 [CYP27A1], CYP27B1, CYP24A1, CYP2R1, and retinoid X receptor α) was available for 496 fatal cases and 3577 controls. Unconditional logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for the associations of 25(OH)D and SNPs with fatal PCa. The study also tested for 25(OH)D-SNP interactions among 264 fatal cases and 1169 controls. No statistically significant relationship was observed between 25(OH)D and fatal PCa (OR for extreme quartiles, 0.86; 95% CI, 0.65-1.14; P for trend = .22) or the main effects of the SNPs and fatal PCa. There was evidence suggesting that associations of several SNPs, including 5 related to circulating 25(OH)D, with fatal PCa were modified by 25(OH)D. Individually, these associations did not remain significant after multiple testing; however, the P value for the set-based test for CYP2R1 was .002. Statistically significant associations were not observed for either 25(OH)D or vitamin D-related SNPs with fatal PCa. The effect modification of 25(OH)D associations by biologically plausible genetic variation may deserve further exploration.


DNA origami demonstrate the unique stimulatory power of single pMHCs as T cell antigens.
T cells detect with their T cell antigen receptors (TCRs) the presence of rare agonist peptide/MHC complexes (pMHCs) on the surface of antigen-presenting cells (APCs). How extracellular ligand binding triggers intracellular signaling is poorly understood, yet spatial antigen arrangement on the APC surface has been suggested to be a critical factor. To examine this, we engineered a biomimetic interface based on laterally mobile functionalized DNA origami platforms, which allow for nanoscale control over ligand distances without interfering with the cell-intrinsic dynamics of receptor clustering. When targeting TCRs via stably binding monovalent antibody fragments, we found the minimum signaling unit promoting efficient T cell activation to consist of two antibody-ligated TCRs within a distance of 20 nm. In contrast, transiently engaging antigenic pMHCs stimulated T cells robustly as well-isolated entities. These results identify pairs of antibody-bound TCRs as minimal receptor entities for effective TCR triggering yet validate the exceptional stimulatory potency of single isolated pMHC molecules.


Detection of natural infection with Mycobacterium intracellulare in healthy wild-caught Chacma baboons (Papio ursinus) by ESAT-6 and CFP-10 IFN-gamma ELISPOT tests following a tuberculosis outbreak.
Both tuberculous and non-tuberculous mycobacteria can cause infection in nonhuman primates (NHP), indicating the existence of potential zoonotic transmission between these animals and visitors to zoos or animal handlers in primate facilities. Screening of mycobacterial infections in NHP is traditionally done by tuberculin skin test (TST), which is unable to distinguish between pathogenic and non-pathogenic mycobacterial infections. In this study, we investigated the use of ESAT-6 and CFP-10 for detection of mycobacterial infections in a wild-caught baboon colony after one baboon died of tuberculosis (TB). Peripheral blood lymphocytes for interferon-gamma enzyme-linked immunospot assay (IFN-gamma ELISPOT) assay were obtained from TST positive baboons and those in contact with tuberculous baboons before being euthanased, autopsied and lung tissues taken for histology and mycobacterial culture. Both ESAT-6 and CFP-10 IFN-gamma ELISPOT assays were able to detect early M. tuberculosis but also M. intracellulare infection. Although this indicates potential cross-reactivity with M. intracellulare antigens, the method was able to distinguish M. bovis BCG vaccination from M. tuberculosis infection. This assay performed better than the TST, which failed to detect one M. tuberculosis and two early M. intracellulare infections. These results suggest that the IFN-gamma ELISPOT assay could improve the detection of M tuberculosis infections when screening NHP. There is some doubt, however, concerning specificity, as the assay scored positive three animals infected with M. intracellulare.


Elucidation of inhibitor-binding pockets of d-amino acid oxidase using docking simulation and N-sulfanylethylanilide-based labeling technology.
Because of the relevance of d-serine (d-Ser) to schizophrenia, inhibitors of d-amino acid oxidase (DAO), which catalyzes degradation of d-Ser in the presence of flavin adenine dinucleotide (FAD), are expected to be anti-schizophrenia therapeutics. In this study, binding pockets of DAO to its inhibitor 4-bromo-3-nitrobenzoic acid were searched by combining in silico docking simulation and labeling experiments employing an N-sulfanylethylanilide-based labeling technology that we have developed. The results clearly demonstrated that there are two binding pockets: one is shared with d-Ser and FAD, and the other is an unexpected cleft between the subunits of a DAO dimer. These findings will provide insight to aid the development of new DAO inhibitors. In addition, it was also proved that our labeling technology could be applicable to elucidate the binding pockets of proteins.


Perceptions of female teenagers in the Tshwane District on the use of contraceptives in South Africa.
Perceptions of female teenagers in the Tshwane District contribute to the nonuseand or discontinued use of contraceptives as evidenced by increased levels of unplanned pregnancies. The objective of this study was to explore and describe the perceptions of female teenagers in the Tshwane District on the use of contraceptives. A qualitative, explorative, descriptive approach was followed in this study. The population comprised of pregnant female teenagers who were purposively selected. Data were collected using unstructured individual interviews on a face-to-face encounter in a natural setting. Data were analysed using the discourse method of data analysis. The following perceptions on the use of contraceptives emerged: Perceptions on the use of contraceptives, emotions, contraceptive effects, social pressure and education on contraceptives. Teenagers' perceptions were predominantly negative with unfounded fears. Though the teenagers were aware of the importance of the use of contraceptives, motivation to pursue contraception was lacking. Teenagers verbalised to be uncommitted as well. Various perceptions of female teenagers in the Tshwane District on the use of contraceptives were explored and described. It was noted that all the teenagers interviewed had great remorse and feelings of guilt regarding their behaviour of not using contraceptives.Their need for re-education was cited and seen as motivational enough to encourage the use of contraceptives at primary health care settings. Therefore, the study recommended that health education programmes should be restructured to effectively influence the female teenagers'perceptions positively and to promote the use of contraceptives.


Clustering nuclear receptors in liver regeneration identifies candidate modulators of hepatocyte proliferation and hepatocarcinoma.
Liver regeneration (LR) is a valuable model for studying mechanisms modulating hepatocyte proliferation. Nuclear receptors (NRs) are key players in the control of cellular functions, being ideal modulators of hepatic proliferation and carcinogenesis. We used a previously validated RT-qPCR platform to profile modifications in the expression of all 49 members of the NR superfamily in mouse liver during LR. Twenty-nine NR transcripts were significantly modified in their expression during LR, including fatty acid (peroxisome proliferator-activated receptors, PPARs) and oxysterol (liver X receptors, Lxrs) sensors, circadian masters RevErbα and RevErbβ, glucocorticoid receptor (Gr) and constitutive androxane receptor (Car). In order to detect the NRs that better characterize proliferative status vs. proliferating liver, we used the novel Random Forest (RF) analysis to selected a trio of down-regulated NRs (thyroid receptor alpha, Trα; farsenoid X receptor beta, Fxrβ; Pparδ) as best discriminators of the proliferating status. To validate our approach, we further studied PPARδ role in modulating hepatic proliferation. We first confirmed the suppression of PPARδ both in LR and human hepatocellular carcinoma at protein level, and then demonstrated that PPARδ agonist GW501516 reduces the proliferative potential of hepatoma cells. Our data suggest that NR transcriptome is modulated in proliferating liver and is a source of biomarkers and bona fide pharmacological targets for the management of liver disease affecting hepatocyte proliferation.


Asynchrony in discrete bimanual aiming: evidence for visual strategies of coordination.
The bimanual coupling literature supposes an inherent drive for synchrony between the upper limbs when making discrete bimanual movements. The level of synchrony is argued to be task dependent, reliant on the visual demands of the two targets, and the result of a complex pattern of hand and eye movements (Bingham, Hughes, & Mon-Williams, 2008 ; Riek, Tresilian, Mon-Williams, Coppard, & Carson, 2003). However, recent work by Bruyn and Mason (2009) suggests that temporal coordination is not solely influenced by visual saccades. In this experimental series, a total of 8 participants performed congruent movements to targets either near or far from the midline. Targets far from the midline, requiring a visual saccade, resulted in greater terminal asynchrony. Initial and terminal asynchrony were not consistent, but linked to the task demands at that stage of the movement. If the asynchrony evident at the end of a bimanual movement is due to a complex pattern of hand and eye movements then the removal of visual feedback should result in an increase in synchrony. Sixteen participants then completed congruent and incongruent bimanual aiming movements to near and/or far targets. Movements were made with or without visual feedback of hands and targets. Analyses revealed that movements made without visual feedback showed increased synchrony between the limbs, yet movements to incongruent targets still showed greater asynchrony. We suggest that visual constraints are not the sole cause of asynchrony in discrete bimanual movements.


Herpes zoster post-herpetic neuralgia.
Post-herpetic neuralgia (PHN) is the most frequent complication of herpes zoster and often results in significant morbidity and a reduction in the patient's quality of life. The peripheral nerve injury that occurs during the acute phase of herpes zoster (HZ) leads to an abnormal tonic impulse discharge from primary nociceptive afferent neurons which induce slow temporal summation. This "wind-up" phenomenon is responsible for continuous partial depolarisation of second-order neurons with increased spontaneous impulse discharge and expanded receptive fields within the dorsal horn nociceptive neurons. The abnormal central processing involves the activation of N-methyl-D-aspartate (NMDA) receptors resulting in neuropathic pain, characterized by spontaneous pain, hyperalgesia and allodynia which is typical of PHN. In addition, tonic input from non-nociceptive AB afferent neurons, maintained by sympathetic efferent activity, contribute to the development and maintenance of neuropathic pain in general, and a burning sensation in particular.


The impact of structural variation on human gene expression.
Structural variants (SVs) are an important source of human genetic diversity, but their contribution to traits, disease and gene regulation remains unclear. We mapped cis expression quantitative trait loci (eQTLs) in 13 tissues via joint analysis of SVs, single-nucleotide variants (SNVs) and short insertion/deletion (indel) variants from deep whole-genome sequencing (WGS). We estimated that SVs are causal at 3.5-6.8% of eQTLs-a substantially higher fraction than prior estimates-and that expression-altering SVs have larger effect sizes than do SNVs and indels. We identified 789 putative causal SVs predicted to directly alter gene expression: most (88.3%) were noncoding variants enriched at enhancers and other regulatory elements, and 52 were linked to genome-wide association study loci. We observed a notable abundance of rare high-impact SVs associated with aberrant expression of nearby genes. These results suggest that comprehensive WGS-based SV analyses will increase the power of common- and rare-variant association studies.


Increased rates of adverse perinatal outcomes in women with gestational diabetes and depression.
We sought to examine the impact of depression on adverse perinatal outcomes in women with Gestational Diabetes Mellitus (GDM). We performed a retrospective cohort study comparing the rates of perinatal complications among singleton, nonanomalous births to women with GDM and the diagnosis of depression compared to GDM women without depression between 2007 and 2011 in California. Perinatal outcomes were analyzed using chi-square and multivariable logistic regression to compare frequencies of characteristics and outcomes and to determine the strength of association of depression and adverse perinatal outcomes among women with GDM. Statistical comparisons with a p-value of less than .05 and 95% CI that did not cross the null were considered statistically significant. Among the cohort of 170,572 women with GDM, 2090 (1.22%) were diagnosed with antenatal depression. Women with GDM and depression had significantly higher rates of preeclampsia (adjusted Odds Ratio [aOR] 1.28, 95% CI 1.11-1.49) and gestational hypertension (aOR 1.23, 95% CI 1.05-1.44). Women with GDM and depression also had higher rates of preterm delivery at <37, and <34 weeks gestational age (aOR 1.33, 95% CI 1.18-1.50 and 1.36, 95% CI 1.15-1.61, respectively). Women with GDM and a diagnosis of depression have higher rates of adverse perinatal outcomes than women with GDM alone. Identifying and managing depression among women with GDM has the potential to improve the care and health of this high-risk population.


Carotid Atherosclerotic Plaque Characteristics on Magnetic Resonance Imaging Relate With History of Stroke and Coronary Heart Disease.
Because atherosclerosis is a systemic disease, presence and composition on 1 location may relate to ischemic events in distant locations. We examined whether carotid atherosclerotic wall thickness, stenosis, and plaque composition are related to history of ischemic stroke and coronary heart disease (CHD). From the population-based Rotterdam Study, 1731 asymptomatic participants (mean age, 72.4±9.1 years; 55% males) underwent magnetic resonance imaging of both carotid arteries. We assessed carotid wall thickness, stenosis and plaque composition, that is presence of intraplaque hemorrhage, lipid, and calcification. History of ischemic stroke and CHD was assessed until date of magnetic resonance imaging. The study was approved by the institutional review board, and all participants gave informed consent. Logistic regression analyses adjusted for age and traditional cardiovascular risk factors were used to study sex-specific associations between plaque characteristics and clinical events. We found that both carotid stenosis and intraplaque hemorrhage were associated with ischemic stroke in men but not in women (men: odds ratio [OR] for stenosis [per 10% increase]: 1.17 [95% CI, 1.06-1.30] and for intraplaque hemorrhage 2.39 [95% CI, 1.32-4.35]). In both men and women, carotid stenosis was associated with CHD (men: OR per 10% increase 1.12 [95% CI, 1.04-1.21] and women: OR, 1.17 [95% CI, 1.03-1.34]) and carotid wall thickness was associated with CHD (men: OR, 1.20 [95% CI, 1.03-1.39] and women: OR, 1.21 [95% CI, 0.88-1.65]). None of the plaque components was associated with CHD. Whereas carotid plaque thickness and stenosis are associated with the history of ischemic stroke and CHD, carotid intraplaque hemorrhage is associated with ischemic stroke, but not with CHD, providing novel insights into the pathogenesis of cardiovascular events.


Is uric acid an indicator of metabolic syndrome in the first-degree relatives of patients with type 2 diabetes?
To determine whether uric acid levels are associated with the components of metabolic syndrome and whether uric acid is a significant factor for development of metabolic syndrome in the first-degree relatives of type 2 diabetic patients as high risk group. A total of 694 (182 male and 512 female, aged 30-69 years) first-degree relatives of type 2 diabetic patients during 2007-2011 were enrolled. The height, weight, waist circumference, blood pressure, fasting plasma glucose, lipid profile and uric acid concentrations were measured. Metabolic syndrome was defined by NCEP-ATP III. Uric acid was associated with waist circumference, blood pressure, triglyceride and HDL-cholesterol level in both sexes (r = 0.1-0.3, P < 0.05). The prevalence of metabolic syndrome in the fourth quartile of uric acid (64.4% of male and 60.2% of female population) was significantly more than those in the first (25.5% of male and 31.2% of female population) and second quartiles (33.3% of male and 32.0% of female population). The mean of uric acid in people with metabolic syndrome was significantly higher than in those without (6.6 ± 1.2 mg/dL vs. 5.8 ± 1.2 mg/dL; P = 0.0001). The age-adjusted odds ratios (95% confidence interval) of uric acid for metabolic syndrome in univariate analysis were [1.60 (1.23-2.07); P = 0.008] for men and [1.61 (1.34-1.92); P = 0.0001] for women but the effect of uric acid in multivariate logistic regression was not significant. Uric acid is associated with majority of the metabolic syndrome components. People with metabolic syndrome have higher uric acid levels. However, uric acid probably is not an independent factor to predict the metabolic syndrome.


Development and survival of infective larvae of nematode parasites of sheep on pasture in a cool tropical environment.
The development and survival of nematode parasites of sheep were studied in a cool tropical environment of the highlands of Ethiopia on 24 plots serially contaminated with Haemonchus contortus, Longistrongylus elongata and Trichostrongylus colubriformis eggs over a period of 2 consecutive years from June 1993 to May 1995. The availability of infective larvae was monitored by monthly pasture sampling and larvae recovery. Infective larvae were recovered from the herbage collected from the plots contaminated in June, July, August, September and October of both years. The longevity of infective larvae varied between two and six weeks when eggs were deposited on pasture in June and October, respectively. During the dry and short rainy season (November through May), eggs failed to develop into L3 stage. It is suggested that the long period (7 to 8 months) of lack of development of trichostrongylid infective larvae on the pasture can be efficiently used in a strategic treatment programme to interrupt transmission of H. contortus, L. elongata and T. colubriformis in sheep in this environment.


Alternative human eIF5A protein isoform plays a critical role in mitochondria.
The eukaryotic translation initiation factor 5A (eIF5A) is the only known protein containing the amino acid residue hypusine, essential for its activity. Hypusine residue is produced by a posttranslational modification involving deoxyhypusine synthetase and deoxyhypusine hydroxylase. Herein, we aimed to describe the role of the alternative human isoform A on mitochondrial processes. Isoform A depletion modulates oxidative metabolism in association with the downregulation of mitochondrial biogenesis-related genes. Through positive feedback, it increases cell respiration leading to highly reactive oxygen species production, which impacts mitochondrial bioenergetics. These metabolic changes compromise mitochondrial morphology, increasing its electron density and fission, observed by transmission electron microscopy. This set of changes leads the cells to apoptosis, evidenced by increased DNA fragmentation and proapoptotic BAK protein content increase. Thus, we show that the alternative eIF5A isoform A is crucial for energy metabolism controlled by mitochondria and cellular survival.


Three kidneys, two diseases, one antibody?
Anti-factor H antibody has been recently described as responsible for thrombotic microangiopathies (TMA) as well as membranoproliferative glomerulonephritis (MPGN). We report here, for the first time, the case of a woman with an anti-factor H antibody, who developed MPGN on native kidney, rapid recurrence on first graft, and TMA on second graft despite immunosuppressive therapy and plasma exchanges. This case supports the hypothesis that MPGN and TMA are closely linked by common pathogenic mechanisms and the need for complete exploration of complement pathway including factor H activity and autoantibody in front of any MPGN.


cis-active structural motifs involved in specific encapsidation of Moloney murine leukemia virus RNA.
We have analyzed the roles of RNA structural motifs located in the 5' part of the Moloney murine leukemia virus (M-MuLV) encapsidation domain (Psi region) with regard to their effects on viral replication. Four putative stem-loop structures between the 5' splice donor site and the gag initiation codon have been examined: stem structure A, corresponding to M-MuLV viral nucleotides 211 to 224; stem-loop B, nucleotides 278 to 303; stem-loop C, nucleotides 310 to 352; and stem-loop D, nucleotides 355 to 374. By measuring infectivities, encapsidation and splicing efficiencies, and endogenous reverse transcription levels of motif A, B, C, and D deletion mutants, we identified mutations which affect replication at the encapsidation step. In particular, deletion of all four motifs in a single mutant eliminated encapsidation of viral RNA, while deletion of individual elements moderately reduced the encapsidation efficiencies. Through analysis of different deletion combinations, we found that deletion of the first two motifs (A plus B) reduced both encapsidation and reverse transcription efficiencies, while deletion of the 3' motifs (C plus D) eliminated encapsidation. Interestingly, the C and D motifs both contain a GACG loop sequence and are highly conserved among murine type C retroviruses. Our results indicate that M-MuLV motifs C and D are necessary for efficient encapsidation, and the presence of at least one of these two stem-loops is crucial to encapsidation and virus replication.


Early trauma-focused cognitive-behavioural therapy to prevent chronic post-traumatic stress disorder and related symptoms: a systematic review and meta-analysis.
Early trauma-focused cognitive-behavioural therapy (TFCBT) holds promise as a preventive intervention for people at risk of developing chronic post-traumatic stress disorder (PTSD). The aim of this review was to provide an updated evaluation of the effectiveness of early TFCBT on the prevention of PTSD in high risk populations. We performed a systematic literature search in international electronic databases (MEDLINE, EMBASE, PsycINFO, CENTRAL, CINAHL, ISI and PILOTS) and included randomised controlled trials comparing TFCBT delivered within 3 months of trauma, to alternative interventions. All included studies were critically appraised using a standardised checklist. Two independent reviewers selected studies for inclusion and assessed study quality. Data extraction was performed by one reviewer and controlled by another. Where appropriate, we entered study results into meta-analyses. Seven articles reporting the results of five RCTs were included. All compared TFCBT to supportive counselling (SC). The study population was patients with acute stress disorder (ASD) in four trials, and with a PTSD diagnosis disregarding the duration criterion in the fifth trial. The overall relative risk (RR) for a PTSD diagnosis was 0.56 (95% CI 0.42 to 0.76), 1.09 (95% CI 0.46 to 2.61) and 0.73 (95% CI 0.51 to 1.04) at 3-6 months, 9 months and 3-4 years post treatment, respectively. A subgroup analysis of the four ASD studies only resulted in RR = 0.36 (95% CI 0.17 to 0.78) for PTSD at 3-6 months. Anxiety and depression scores were generally lower in the TFCBT groups than in the SC groups. There is evidence for the effectiveness of TFCBT compared to SC in preventing chronic PTSD in patients with an initial ASD diagnosis. As this evidence originates from one research team replications are necessary to assess generalisability. The evidence about the effectiveness of TFCBT in traumatised populations without an ASD diagnosis is insufficient.


The distribution of faecal nematode egg counts in Scottish Blackface lambs following natural, predominantly Ostertagia circumcincta infection.
Faecal samples were taken in 2 consecutive years from Scottish Blackface lambs on a commercial farm in central Scotland to examine variation among lambs in the number of nematode eggs in their faeces. Samples were taken at intervals of 4 weeks from 1 to 5 months of age. Lambs were treated with a broad-spectrum anthelmintic at each sample date. Ewes and lambs were naturally infected by grazing pasture contaminated with mixed, predominantly Ostertagia circumcincta, nematode species. The ewes were removed from the lambs when the lambs were three months old. Only 3 of the 6 sets of samples taken prior to removal of the ewes fitted a negative binomial distribution but all sets of samples taken after this did so. The mean and the variance differed with the month and year of sampling. The amount of variation among lambs increased as the lambs grew from 3 to 5 months of age, possibly because of heterogeneity in the rate of development of protective responses. Counts taken in August and September, when the lambs were 4 and 5 months of age, were positively correlated with each other. In contrast, counts taken at 1 or 2 months of age were negatively correlated with counts taken when the lambs were 3-5 months old. Sex, date of birth, sire and dam were all associated with variation in faecal egg counts but the size and significance of their influence was dependent upon the date when faecal samples were taken. Four factors: sire, dam, sex and year accounted for over two-thirds of the variation among older lambs in faecal egg counts.


Tumour necrosis factor-alpha (TNF-alpha) in patients who have asbestosis and develop cancer.
Concentrations of tumour necrosis factor-alpha (TNF-alpha) were assayed by radioimmunoassay in serum samples collected between 1981 and 1987 from 111 patients with asbestosis who were at a high risk of cancer. Follow up of these patients until 1993 showed that 38 had developed cancer (27 lung, three mesotheliomas, and eight diverse malignancies). The mean serum concentrations of TNF-alpha given in fmol/100 microliters serum in all the cases with cancer (14.1) and the cases with lung cancer (13.6) were significantly higher (P < 0.05) than the mean concentrations in the exposed controls (10.5). A positive increase was considered to be any value that was > 2 SDs above the mean of the exposed controls. 22% (six of 27) of the cases with lung cancer were positive compared with 4% (three of 73) of the exposed controls, a significant difference (P < 0.001). The serum concentrations of TNF-alpha correlated moderately with cancer (r = 0.3), lung cancer (r = 0.3), and Neu oncoproteins and epidermal growth factor receptor (EGFR) (r = 0.3, 0.5 respectively). Also, there was a significant correlation between development of cancer and severity or progression of asbestosis. There was no correlation between the concentrations of TNF-alpha and severity or progression of asbestosis. These results showed high concentrations of TNF-alpha in the patients who had cancer. TNF-alpha may offer an auxiliary method in early diagnosis of cancers related to asbestosis.


Gender difference in disease expression, radiographic damage and disability among patients with psoriatic arthritis.
The authors aimed to assess gender-related differences in severity of psoriatic arthritis (PsA) as reflected by measures of disease activity, joint damage, quality of life and disability. A cross-sectional analysis was performed among patients who have been followed in a large PsA clinic. Demographic, clinical and radiographic data as well as information about quality of life and function were retrieved from the clinic database. Radiographic damage was assessed according to modified Steinbrocker score (mSS). The association between gender and the following outcome variables, radiographic joint damage, axial involvement and measures of quality of life and function, was assessed by multivariate regression analysis after adjustment for potential confounders. Three hundred and forty-five men and 245 women were included in the study. Axial involvement was more frequent in men (42.9% vs 31%, p=0.003). In multivariate analysis, adjusting for potential confounders, men were more likely to develop axial involvement (OR 1.8, p=0.003). Men were also more likely to develop more severe radiographic damage in the peripheral joints as evident by mSS. Men were more likely to be in a higher mSS damage category compared with women after adjusting for potential confounders in multivariate analysis (OR 1.6, p=0.007). Women suffered from more severe limitations in function and worse quality of life compared with men based on several patients' reported outcomes. Men with PsA are more likely to develop axial involvement and radiographic joint damage, while women are more likely to report about limitation in function and impaired quality of life.


Correlation between the binding affinity and the conformational entropy of nanobody SARS-CoV-2 spike protein complexes.
Camelid single-domain antibodies, also known as nanobodies, can be readily isolated from naïve libraries for specific targets but often bind too weakly to their targets to be immediately useful. Laboratory-based genetic engineering methods to enhance their affinity, termed maturation, can deliver useful reagents for different areas of biology and potentially medicine. Using the receptor binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein and a naïve library, we generated closely related nanobodies with micromolar to nanomolar binding affinities. By analyzing the structure-activity relationship using X-ray crystallography, cryoelectron microscopy, and biophysical methods, we observed that higher conformational entropy losses in the formation of the spike protein-nanobody complex are associated with tighter binding. To investigate this, we generated structural ensembles of the different complexes from electron microscopy maps and correlated the conformational fluctuations with binding affinity. This insight guided the engineering of a nanobody with improved affinity for the spike protein.


An epidemic of sylvatic yellow fever in the southeast region of Maranhao State, Brazil, 1993-1994: epidemiologic and entomologic findings.
Yellow fever virus transmission was very active in Maranhao State in Brazil in 1993 and 1994. An investigation was carried out to evaluate the magnitude of the epidemic. In 1993, a total of 932 people was examined for yellow fever from Maranhao: 70 were positive serologically, histopathologically, and/or by virus isolation, and another four cases were diagnosed clinically and epidemiologically. In Mirador (17,565 inhabitants), the incidence was 3.5 per 1,000 people (case fatality rate [number of deaths/number of cases diagnosed] = 16.4%), while in a rural yellow fever risk area (14,659 inhabitants), the incidence was 4.2 and the case-fatality rate was 16.1% (10 of 62). A total of 45.2% (28 of 62) asymptomatic infections were registered. In 1994, 49 serum samples were obtained and 16 cases were confirmed (two by virus isolation, two by seroconversion, and 12 by serology). No fatal cases were reported. In 1993, 936 potential yellow fever vectors were captured in Mirador and a single strain was isolated from a pool of Haemagogus janthinomys (infection rate = 0.16%). In 1994, 16 strains were isolated from 1,318 Hg. janthinomys (infection rate = 1.34%) and one Sabethes chloropterus (infection rate = 1.67%). Our results suggest that this was the most extensive outbreak of yellow fever in the last 20 years in Brazil. It is also clear that the lack of vaccination was the principal reason for the epidemic, which occurred between April and June, during the rainy season, a period in which the mosquito population in the forest increases.


ASCO 2016: highlights in breast cancer.
At the 2016 ASCO Annual Meeting, several pertinent studies in the field of breast cancer were presented. MA17.R was the first randomized phase III trial to evaluate the prolongation of adjuvant aromatase-inhibitor (AI) therapy from 5 to 10 years; while a significant reduction of disease-free survival events was observed in the extended treatment group, the absolute difference was relatively small and longer endocrine therapy resulted in a higher fracture rate. A combined analysis of three North American trials emphasized the superiority of anthracycline containing adjuvant chemotherapy regimens compared with docetaxel/cyclophosphamide (TC), while the PANTHER trial investigated dose-dense tailored adjuvant treatment. In metastatic breast cancer, the main interest was on cyclin-dependent kinase (CDK) 4/6 inhibitors. In PALOMA-2, the addition of palbociclib to letrozole prolonged progression-free survival (PFS) from 14.5 to 24.8 months resulting in the longest PFS data ever reported in the first-line setting. A subgroup analysis of premenopausal patients accrued to PALOMA-3 indicated that in this patient subset, ovarian function suppression plus fulvestrant and palbociclib yielded results comparable to the postmenopausal population. ESR1 mutations were another focus of interest as these activating mutations in the gene coding for the estrogen receptor alpha apparently evolve under the selection pressure of AI therapy.


Variability in the phases of the menstrual cycle.
To determine variability in the phases of the menstrual cycle among healthy, regularly cycling women. A prospective descriptive study of a new data set with biological markers to estimate parameters of the menstrual cycles. One hundred forty one healthy women (mean age 29 years) who monitored 3 to 13 menstrual cycles with an electronic fertility monitor and produced 1,060 usable cycles of data. MEASURES AND OUTCOMES: Variability in the length of the menstrual cycle and of the follicular, fertile, and luteal phases, and menses. The estimated day of ovulation and end of the fertile phase was the peak fertility reading on the monitor (i.e., the urinary luteinizing hormone surge). Mean total length was 28.9 days (SD = 3.4) with 95% of the cycles between 22 and 36 days. Intracycle variability of greater than 7 days was observed in 42.5% of the women. Ninety-five percent of the cycles had all 6 days of fertile phase between days 4 and 23, but only 25% of participants had all days of the fertile phase between days 10 and 17. Among regularly cycling women, there is considerable normal variability in the phases of the menstrual cycle. The follicular phase contributes most to this variability.


Effect of freezing rate on physical stability of lyophilized cationic liposomes.
Factors affecting the storage stability of lyophilized cationic liposomes were investigated using liposomes prepared with various excipients and by different freezing rates, either quick freezing (freezing by immersion into liquid nitrogen) or slow freezing (cooling to -50 degrees C at a rate of -10 degrees C/h). Increases in the particle size of cationic liposomes observed during freeze-drying were inhibited by the addition of sucrose, trehalose and sucrose-dextran mixtures (1 : 1 and 2 : 1 by weight). The storage instability of the formulations, as indicated by changes in particle size, was affected by their glass transition temperature (T(g)). Addition of high-T(g) excipients resulted in smaller increases in the particle size, indicating improvement of storage stability. The storage stability of cationic liposome formulations was also affected by freezing rate. Formulations prepared by slow freezing exhibited better stability. Longer shear relaxation times were observed for formulations prepared by slow freezing compared with those prepared by quick freezing. This indicates that formulations prepared by slow freezing have a lower matrix mobility, which may result in better storage stability. T(g) or (1)H-NMR relaxation measurements could not detect differences in matrix mobility between formulations prepared by different freezing rates. Shear relaxation measurements seem to be a useful method for evaluating the storage stability of cationic liposome formulations.


China's rapidly aging population creates policy challenges in shaping a viable long-term care system.
In China, formal long-term care services for the large aging population have increased to meet escalating demands as demographic shifts and socioeconomic changes have eroded traditional elder care. We analyze China's evolving long-term care landscape and trace major government policies and private-sector initiatives shaping it. Although home and community-based services remain spotty, institutional care is booming with little regulatory oversight. Chinese policy makers face mounting challenges overseeing the rapidly growing residential care sector, given the tension arising from policy inducements to further institutional growth, a weak regulatory framework, and the lack of enforcement capacity. We recommend addressing the following pressing policy issues: building a balanced system of services and avoiding an "institutional bias" that promotes rapid growth of elder care institutions over home or community-based care; strengthening regulatory oversight and quality assurance with information systems; and prioritizing education and training initiatives to grow a professionalized long-term care workforce.


Tracking health care service use and the experiences of adults with autism spectrum disorder without intellectual disability: A longitudinal study of service rates, barriers and satisfaction.
Adults with Autism Spectrum Disorder (ASD) encounter many difficulties finding and accessing health care services. Despite this, few studies have considered the health service use patterns of adults with ASD without intellectual disability (ID). The current study examines a diverse range of medical and mental health services and supports, as well as adults' personal experiences accessing and using these services, barriers to service use, and reported unmet service needs. Forty adults (ages 18-61 years) with ASD without ID completed surveys every two months about their health service use for a total of 12-18 months. Bivariate analyses were conducted to understand the individual demographic and clinical factors associated with rate of service use, satisfaction with services, and barriers to health care. Results indicated that, beyond a family doctor, the most commonly used services were dentistry, individual counseling, and psychiatry. Individuals who had medical problems experienced significantly more barriers to service use than those who did not, and those who had medical and mental health problems were less satisfied with services. Findings highlight the challenges adults with ASD without ID face accessing appropriate, quality services to meet their needs, particularly those with complex medical and mental health issues. Service providers must strive to provide adequate health care to this population who may become distressed if their needs are left unmet.


Excess of counterclockwise scalp hair-whorl rotation in homosexual men.
While most men prefer women as their sexual partners, some are bisexual and others are homosexuals. It has been debated for a long time whether a person's sexual preference is innate, learned, or due to a combination of both causes. It was recently discovered that the human right-versus-left-hand use preference and the direction of scalp hair-whorl rotation develop from a common genetic mechanism. Such a mechanism controls functional specialization of brain hemispheres. Whether the same mechanism specifying mental makeup influences sexual preference was determined here by comparing hair-whorl rotation in groups enriched with homosexual men with that in males at large. Only a minority of 8.2% (n = 207) unselected 'control' group of males had counterclockwise rotation. In contrast, all three samples enriched with homosexual men exhibited highly significant (P < 0.0001), 3.6-fold excess (29.8%, n = 272) counterclockwise rotation. These results suggest that sexual preference may be influenced in a significant proportion of homosexual men by a biological/genetic factor that also controls direction of hair-whorl rotation.


Seeps regulate stream nitrate concentration in a forested Appalachian catchment.
Surface seeps can be defined as locations where upwelling ground water saturates the surface for most of the year and excess ground water can be delivered to the stream channel via surface flowpaths. If a stream is predominantly fed by seeps, then ground water added to the stream via these surface flowpaths may result in reduced interactions with the subsurface riparian zone. It is generally believed that seep ground water that upwells and then flows along surface flowpaths can be subject to diminished denitrification and biologic uptake processes. Seep effects on stream nitrate (NO(3)) concentration were studied in Baldwin Creek (5.35 km(2)), southwestern Pennsylvania. Nitrate retention within seep zones was evaluated over a 1-yr period (May 2002-2003) using a monthly, nested (top and bottom of seep) sampling approach along 15 individual seeps. Seep samples were analyzed for NO(3)-N, NH(3)-N, and dissolved organic carbon, along with stream waters and streamflow measurements at seven stream stations. Seeps were generally NO(3) sinks with concentrations decreasing downseep: 31% median annual reduction and 73% maximum monthly reduction. During cold and wet periods, seeps frequently behaved as NO(3) sources to the stream (NO(3) concentrations increased or remained constant downseep). Seep temperature and discharge were related to seasonal variability in seep NO(3) retention. Seasonal variations in stream NO(3) concentration have been attributed to upland soil and vegetation processes in numerous watersheds. At Baldwin Creek, seep NO(3) processing regulated the seasonal variability of stream NO(3) concentrations. These results suggest that seeps provide important water quality functions and can modulate the effects of elevated regional N deposition in Appalachian catchments.


Intralobular heterogeneity of carbon tetrachloride-induced oxidative stress in perfused rat liver visualized by digital imaging fluorescence microscopy.
Spatial and temporal alterations in carbon tetrachloride-induced oxidative stress were studied in perfused hepatic microcirculatory units of the rat by digital imaging microscopic fluorography using dichlorofluorescin diacetate, a hydroperoxide-sensitive fluorogenic probe. The surface of the liver loaded with dichlorofluorescin was microscopically visualized, and the fluorescence of dichlorofluorescin (DCF), a highly fluorescent molecule generated by hydroperoxide-mediated oxidation of dichlorofluorescin, was digitally processed. After completing the experiments, fluorescein-labeled albumin was injected into the perfusate to confirm the state of sinusoidal perfusion and the topography of hepatic microangioarchitecture in the area studied. During transportal infusion of carbon tetrachloride, DCF fluorescence was observed predominantly in the pericentral area and was attenuated by pretreatment with SKF-525A, suggesting the involvement of cytochrome P-450. After peaking at the maximum level, the pericentral DCF fluorescence gradually decreased in parallel to the loss of viability, implicating the causal role of intracellular hydroperoxide formation in hepatocellular injury. In retrogradely perfused liver, in which intralobular O2 gradient was reversed, no significant activation of DCF fluorescence was observed among hepatic lobules. It is therefore conceivable that the zonal heterogeneity of carbon tetrachloride-induced lipid peroxidation may depend on at least two factors, sinusoidal oxygenation and the intralobular distribution of cytochrome P-450. Furthermore, development of the current technique will provide a useful method to investigate the microtopography of oxidative stress in organ microcirculatory units.


Retrograde spread of hydrocortisone containing foam given intrarectally in ulcerative colitis.
A method is described of estimating retrograde spread through the colon of a 10% hydrocortisone acetate foam by labelling the foam with technetium-99m sulphur colloid and observing spread after intrarectal administration by serial gamma-camera pictures. The recommended 51 ml dose of foam reached the mid-sigmoid colon in all of the nine patients who had active ulcerative colitis. Furthermore, in seven the foam reached the proximal sigmoid colon. Foam spread less extensively in five patients with quiescent disease than in those with active disease. Increasing the volume of enema to 50 ml did not improve retrograde spread through the colon. These results suggest that 10% hydrocortisone foam may be useful in treating patients with distal ulcerative colitis that is not necessarily limited to the rectum.


LEAPing through the looking glass: secondary analysis of the effect of skin test size and age of introduction on peanut tolerance after early peanut introduction.
In the Learning Early About Peanut Allergy (LEAP) study, early peanut introduction in high-risk 4- to 11-month-olds was associated with a significantly decreased risk of developing peanut allergy. However, the influences of key baseline high-risk factors on peanut tolerance are poorly understood. Secondary analysis was conducted on the publically available LEAP dataset, exploring relationships between peanut tolerance, baseline peanut/egg sensitization, eczema severity/duration, age of introduction, gender, and race. A multiple logistic regression model predicting odds of successful oral food challenge (OFC) at 60 months noted higher odds with early introduction (OR 9.2, P < 0.001, 95% CI 4.2-20.3), white race (OR 2.1, P = 0.04, 95% CI 1.1-3.9), and advancing age (OR 4.8, P = 0.04, 95% CI 1.1-20.8). Odds of peanut tolerance were lower with increasing peanut wheal size (OR 0.58, P < 0.001, 95% CI 0.46-0.74), increased baseline SCORAD score (OR 0.98, P = 0.04, 95% CI 0.97-1), and increased kUA /l of egg serum IgE (sIgE) (OR 0.99, P = 0.04, 95% CI 0.98-1). The probability of peanut tolerance in the early introduction group was 83% vs 43% in the avoidance group with SPT wheal of <4 mm. The probability of a successful OFC was significantly higher with peanut introduction between 6 and 11 months than at 4-6 months. Increasing eczema severity had limited impact on the probability of peanut tolerance in the early introduction arm. Increasing peanut wheal size predicted peanut tolerance only in the avoidance arm. Peanut introduction between 6 and 11 months of age was associated with the highest rates of peanut tolerance, questioning the 'urgency' of introduction before 6 months.


Long Term Clinical and Visual Outcomes of Retrofixated Iris Claw Lenses Implantation in Complicated Cases.
To evaluate the visual outcomes and complications after implantation of retrofixated iris claw (RFIC) lens in various challenging situations. Retrospective, single centre, 8 year clinical audit. A retrospective analysis of cases who underwent RFIC lens implantation alone (group 1) or in combination with vitreoretinal (VR), corneal or glaucoma procedures (group 2) was performed. The main outcomes evaluated were corrected distant visual acuity (CDVA) and postoperative complications. The mean follow up was 13.09±6.8 (range 6-24) months. The study involved 100 eyes of 83 patients with mean age of 51.1±25.4 years. Group 1 included 59 eyes and group 2 had 41 eyes. In group 1, the mean CDVA improved from 0.86±0.81 to 0.38±0.51 LogMAR (p<0.001) with 72.8% eyes having gain in lines (≥ 2 lines) of CDVA with safety index of 1.73. The mean CDVA in group 2 improved from 0.71±0.65 to 0.38±0.34 LogMAR (p=0.003) with 65.8% eyes having gain in lines (≥ 2 lines) of CDVA with a safety index of 1.54. Group 2 showed a higher complication rate of 36.59% compared to group 1 (20.34%). Significant complications noted were secondary glaucoma (8%), disenclavation of haptic (4%), subluxation of RFIC lens (1%). The visual outcome with RFIC lenses when combined with other intraocular procedures is mainly affected by the complexity of co-existing pathologies .The complications are more related to the combined procedures performed rather than RFIC lens implantation alone. This may still be acceptable when complication profile of other intraocular lenses is evaluated in similar challenging situations.


Experiences with the standardized classification of surgical complications (Clavien-Dindo) in general surgery patients.
The standardized Clavien-Dindo classification of surgical complications is applied as a simple and widely used tool to assess and report postoperative complications in general surgery. However, most documentation uses this classification to report surgery-related morbidity and mortality in a single field of surgery or even particular intervention. The aim of the present study was to present experiences with the Clavien-Dindo classification when applied to all patients on the general surgery ward of a tertiary referral care center. We analyzed a period of 6 months of care on a ward with a broad range of general and visceral surgery. Discharge reports and patient charts were analyzed retrospectively and reported complications rated according to the most recent Clavien-Dindo classification version. The complexity of operations was assessed with the Austrian Chamber of Physicians accounting system. The study included 517 patients with 817 admissions, of whom 463 had been operated upon. Complications emerged in 12.5%, of which 19% were rated as Clavien I, 20.7% as Clavien II, 13.8% as Clavien IIIa, 27.6% as Clavien IIIb, 8.6% as Clavien IVa, and 10.3% as Clavien V. No Clavien grade IVb complication occurred within the investigation. Patients having undergone more complex surgery or with higher scores experienced significantly longer lengths of hospital stay. The Clavien-Dindo classification can easily be used to document complication rates in general surgery, even though this collective was not included in the original validation studies of Clavien et al. and consisted of more heavily impaired patients.


Intervention strategies to reduce the burden of soil-transmitted helminths in India.
Soil-transmitted helminth (STH) infections continue to be a major global cause of morbidity, with a large proportion of the burden of STH infections occurring in India. In addition to direct health impacts of these infections, including anaemia and nutritional deficiencies in children, these infections also significantly impact economic development, as a result of delays in early childhood cognitive development and future income earning potential. The current World Health Organization strategy for STH is focused on morbidity control through the application of mass drug administration to all pre-school-aged and school-aged children. In India, the control of STH-related morbidity requires mobilization of significant human and financial resources, placing additional burdens on limited public resources. Infected adults and untreated children in the community act as a reservoir of infection by which treated children get rapidly reinfected. As a result, deworming programmes will need to be sustained indefinitely in the absence of other strategies to reduce reinfection, including water, hygiene and sanitation interventions (WASH). However, WASH interventions require sustained effort by the government or other agencies to build infrastructure and to promote healthy behavioural modifications, and their effectiveness is often limited by deeply entrenched cultural norms and behaviours. Novel strategies must be explored to provide a lasting solution to the problem of STH infections in India other than the indefinite provision of deworming for morbidity control.


Cyclodextrin enhanced biodegradation of polycyclic aromatic hydrocarbons and phenols in contaminated soil slurries.
This work aimed to evaluate the relative contribution of soil catabolic activity, contaminant bioaccessibility, and nutrient levels on the biodegradation of field-aged polycyclic aromatic hydrocarbons and phenolic compounds in three municipal gas plant site soils. Extents of biodegradation achieved, in 6 week-long soil slurry assays, under the following conditions were compared: (i) with inoculation of catabolically active PAH and phenol-degrading microorganisms, (ii) with and without hydroxypropyl-beta-cyclodextrin supplementation (HPCD; 100 g L(-1)), and finally (iii) with the provision of additional inorganic nutrients in combination with HPCD. Results indicated no significant (p < 0.05) differences between biodegradation endpoints attained in treatments inoculated with catabolically active microorganisms as compared with the uninoculated control. Amendments with HPCD significantly (p < 0.05) lowered biodegradation endpoints for most PAHs and phenolic compounds. Only in one soil did the combination of HPCD and nutrients consistently achieve better bioremediation endpoints with respect to the HPCD-only treatments. Thus, for most compounds, biodegradation was not limited by the catabolic activity of the indigenous microorganisms but rather by processes resulting in limited availability of contaminants to degraders. It is therefore suggested that the bioremediation of PAH and phenol impacted soils could be enhanced through HPCD amendments. In addition, the biodegradability of in situ and spiked (deuterated analogues) PAHs following 120 days aging of the soils suggested that this contact time was not sufficient to obtain similar partitions to that observed for field-aged contaminants; with the spiked compounds being significantly (p < 0.05) more available for biodegradation.


Rapid-inflation intermittent pneumatic compression for prevention of deep venous thrombosis.
Current treatment regimens that are designed to prevent deep venous thrombosis in patients undergoing orthopaedic procedures rely predominantly on drug prophylaxis alone. The purpose of this randomized clinical study was to evaluate the effectiveness of a mechanical adjunct to chemoprophylaxis that involves intermittent compression of the legs. During a twenty-two month period, 1803 patients undergoing a variety of orthopaedic procedures were prospectively randomized to receive either chemoprophylaxis alone or a combination of chemoprophylaxis and mechanical prophylaxis. Nine hundred and two patients were managed with low-molecular-weight heparin alone, and 901 were managed with low-molecular-weight heparin and intermittent pneumatic compression of the calves for varying time periods. Twenty-four percent of the patients underwent total hip or knee joint replacement. Screening for deep venous thrombosis was performed on the day of discharge with duplex-color-coded ultrasound. In the chemoprophylaxis-only group, fifteen patients (1.7%) were diagnosed with a deep venous thrombosis; three thromboses were symptomatic. In the chemoprophylaxis plus intermittent pneumatic compression group, four patients (0.4%) were diagnosed with deep venous thrombosis; one thrombosis was symptomatic. The difference between the groups with regard to the prevalence of deep venous thrombosis was significant (p = 0.007). In the chemoprophylaxis plus intermittent pneumatic compression group, no deep venous thromboses were found in patients who received more than six hours of intermittent pneumatic compression daily. Venous thrombosis prophylaxis with low-molecular-weight heparin augmented with a device that delivers rapid-inflation intermittent pneumatic compression to the calves was found to be significantly more effective for preventing deep venous thrombosis when compared with a treatment regimen that involved low-molecular-weight heparin alone.


Evaluation of liver function parameters by Tc-99m-GSA using multivariate analysis: a study of 47 clinical cases.
To investigate the correlation between nuclear medicine parameters determined by technetium-99m-DTPA-galactosyl-human serum albumin (Tc-99m-GSA) and liver function tests, canonical correlation analysis was performed. Tc-99m-GSA studies were performed on 47 patients with hepatocellular carcinoma (HCC) who had undergone transcatheter arterial embolization (TAE). The nuclear medicine parameters LU15, HH15 and LHL15, which are results of nuclear imaging tests, were chosen in combination with the following liver function tests: the serum bilirubin level (T.Bil), the serum albumin level (Alb), serum cholinesterase activity (Ch-E), the clearance rate of indocyanine green (KICG), the hepaplastin test (HPT) and the prothrombin time (PT). The canonical correlation coefficient was 0.7345 and the upper tail probability was 0.00167. A significant correlation was observed between the two sets of variables. The high structural coefficients of Ch-E, KICG and HPT indicated a close relationship with the nuclear medicine parameters, supporting the notion that these nuclear medicine parameters are useful for the estimation of liver damage. The structural coefficients of the nuclear medicine parameters were also high, with LU15 being a parameter as useful as both HH15 and LHL15. T.Bil may evaluate a liver function that is not measured by nuclear imaging techniques, so we should take T.Bil results into account before considering TAE.


Medical professionalism across cultures: a challenge for medicine and medical education.
The recognition of medical professionalism as a complex social construct makes context, geographical location and culture important considerations in any discussion of professional behaviour. Medical students, medical educators and practitioners are now much more on the move globally, exposing them to cultural and social attitudes, values and beliefs that may differ from their own traditional perceptions of professionalism. This paper uses the model of the intercultural development continuum and the concept of "cultural fit" to discuss what might transpire when a student, teacher or doctor is faced with a new cultural environment. Using our own experiences as medical educators working abroad and supported by evidence in the literature, we have developed four anecdotal scenarios to highlight some of the challenges that different cultural contexts bring to our current (Western) understanding of professionalism. The scenarios highlight some of the potentially different regional and/or cultural perspectives and nuances of professional behaviours, attitudes or values that many of us either take for granted or find difficult, depending on our training and socio-cultural upbringing. With this paper, we hope to start a long overdue conversation about global professionalism amongst medical educators, identify potential areas for research and highlight a need for medical schools to embrace a "global" approach to how professionalism is embedded in their curricula.


Nano differential scanning fluorimetry for comparability studies of therapeutic proteins.
Differential scanning calorimetry (DSC) has been extensively used in the biopharmaceutical industry to characterize protein thermal stability and domain folding integrity. Recently, nano differential scanning fluorimetry (nanoDSF) has emerged as a powerful tool for thermal stability analysis and studies of protein domain unfolding. Due to increased interests in the qualification of characterization methods, we are in this study presenting the qualification results for the comparability studies of thermal stability analysis using nanoDSF. The results show that nanoDSF is able to detect thermal transition signals for mAbs, BiTE® molecules, and cytokines at a wide concentration range with high precision, clearly indicating that nanoDSF is suitable for characterization including comparability studies of therapeutic proteins. Compared to the current recognized industry standard DSC, the nanoDSF method enables thermal stability analysis over a much wider concentration range, consumes considerably less materials, and provides significantly higher throughput.


Dealing with missing outcome data in meta-analysis.
Missing data result in less precise and possibly biased effect estimates in single studies. Bias arising from studies with incomplete outcome data is naturally propagated in a meta-analysis. Conventional analysis using only individuals with available data is adequate when the meta-analyst can be confident that the data are missing at random (MAR) in every study-that is, that the probability of missing data does not depend on unobserved variables, conditional on observed variables. Usually, such confidence is unjustified as participants may drop out due to lack of improvement or adverse effects. The MAR assumption cannot be tested, and a sensitivity analysis to assess how robust results are to reasonable deviations from the MAR assumption is important. Two methods may be used based on plausible alternative assumptions about the missing data. Firstly, the distribution of reasons for missing data may be used to impute the missing values. Secondly, the analyst may specify the magnitude and uncertainty of possible departures from the missing at random assumption, and these may be used to correct bias and reweight the studies. This is achieved by employing a pattern mixture model and describing how the outcome in the missing participants is related to the outcome in the completers. Ideally, this relationship is informed using expert opinion. The methods are illustrated in two examples with binary and continuous outcomes. We provide recommendations on what trial investigators and systematic reviewers should do to minimize the problem of missing outcome data in meta-analysis.


ECM Cross-Linking Regulates Invadopodia Dynamics.
Invadopodia are membrane protrusions dynamically assembled by invasive cancer cells in contact with the extracellular matrix (ECM). Invadopodia are enriched by the structural proteins actin and cortactin as well as metalloproteases such as MT1-MMP, whose function is to degrade the surrounding ECM. During metastasis, invadopodia are necessary for cancer cell intravasation and extravasation. Although signaling pathways involved in the assembly and function of invadopodia are well studied, few studies address invadopodia dynamics and how the cell-ECM interactions contribute to cell invasion. Using iterative analysis based on time-lapse microscopy and mathematical modeling of invasive cancer cells, we found that cells oscillate between invadopodia presence and cell stasis-termed the "invadopodia state"-and invadopodia absence during cell translocation-termed the "migration state." Our data suggest that β1-integrin-ECM binding and ECM cross-linking control the duration of each of the two states. By changing the concentration of cross-linkers in two-dimensional and three-dimensional cultures, we generate an ECM in which 0-0.92 of total lysine residues are cross-linked. Using an ECM with a range of cross-linking degrees, we demonstrate that the dynamics of invadopodia-related functions have a biphasic relationship to ECM cross-linking. At intermediate levels of ECM cross-linking (0.39), cells exhibit rapid invadopodia protrusion-retraction cycles and rapid calcium spikes, which lead to more frequent MT1-MMP delivery, causing maximal invadopodia-mediated ECM degradation. In contrast, both extremely high or low levels of cross-linking lead to slower invadopodia-related dynamics and lower ECM degradation. Additionally, β1-integrin inhibition modifies the dynamics of invadopodia-related functions as well as the length of time cells spend in either of the states. Collectively, these data suggest that β1-integrin-ECM binding nonlinearly translates small physical differences in the extracellular environment to differences in the dynamics of cancer cell behaviors. Understanding the conditions under which invadopodia can be reduced by subtle environment-targeting treatments may lead to combination therapies for preventing metastatic spread.


Low--resolution vision in a velvet worm (Onychophora).
Onychophorans, also known as velvet worms, possess a pair of simple lateral eyes, and are a key lineage with regard to the evolution of vision. They resemble ancient Cambrian forms, and are closely related to arthropods, which boast an unrivalled diversity of eye designs. Nonetheless, the visual capabilities of onychophorans have not been well explored. Here, we assessed the spatial resolution of the onychophoran Euperipatoides rowelli using behavioural experiments, three-dimensional reconstruction, anatomical and optical examinations, and modelling. Exploiting their spontaneous attraction towards dark objects, we found that E. rowelli can resolve stimuli that have the same average luminance as the background. Depending on the assumed contrast sensitivity of the animals, we estimate the spatial resolution to be in the range 15-40 deg. This results from an arrangement where the cornea and lens project the image largely behind the retina. The peculiar ellipsoid shape of the eye in combination with the asymmetric position and tilted orientation of the lens may improve spatial resolution in the forward direction. Nonetheless, the unordered network of interdigitating photoreceptors, which fills the whole eye chamber, precludes high-acuity vision. Our findings suggest that adult specimens of E. rowelli cannot spot or visually identify prey or conspecifics beyond a few centimetres from the eye, but the coarse spatial resolution that the animals exhibited in our experiments is likely to be sufficient to find shelter and suitable microhabitats from further away. To our knowledge, this is the first evidence of resolving vision in an onychophoran.