Relating domain size distribution to line tension and molecular dipole density in model cytoplasmic myelin lipid monolayers.
We fit the size distribution of liquid-ordered (L(o)) domains measured from fluorescence images of model cytoplasmic myelin monolayers with an equilibrium thermodynamic expression that includes the competing effects of line tension, λ, dipole density difference, Δm, and the mixing entropy. From these fits, we extract the line tension, λ, and dipole density difference, Δm, between the L(o) and liquid-disordered (L(d)) phases. Both λ and Δm decrease with increasing surface pressure, , although λ/Δm(2) remains roughly constant as the monolayer approaches the miscibility surface pressure. As a result, the mean domain size changed little with surface pressure, although the polydispersity increased significantly. The most probable domain radius was significantly smaller than that predicted by the energy alone, showing that the mixing entropy promotes a greater number of smaller domains. Our results also explain why domain shapes are stable; at equilibrium, only a small fraction of the domains are large enough to undergo theoretically predicted shape fluctuations. Monolayers based on the composition of myelin from animals with experimental allergic encephalomyelitis had slightly lower values of λ and Δm, and a higher area fraction of domains, than control monolayers at all . While it is premature to generalize these results to myelin bilayers, our results show that the domain distribution in myelin may be an equilibrium effect and that subtle changes in surface pressure and composition can alter the distribution of material in the monolayer, which will likely also alter the interactions between monolayers important to the adhesion of the myelin sheath.


Investigation of relationship between the D-dimer and ischemia-modified albumin levels with the radiological imaging-based pulmonary embolism severity score in acute pulmonary embolism.
To investigate possible relationship between the D-dimer and ischemia-modified albumin (IMA) levels and radiological imaging-based severity scores in pulmonary embolism (PE) based on two different radiological characteristics; the pulmonary arterial obstruction index (PAOI) and the right ventricle/left ventricle (RV/LV) ratio. In this prospective cohort study, forty-seven patients presenting to the emergency department and definitively diagnosed with PE using spiral computerized tomography (CT) were initially enrolled in the study. Levels of IMA and D-dimer were assessed colorimetrical and immuno-turbidimetric methods, respectively. The PAOI and RV/LV ratios were calculated from CT images. The levels of biochemical parameters between the groups were compared with use of Mann-Whitney U and Kruskal-Wallis tests and relationship between the radiological scores were assessed using the Spearman correlation test. Analysis of the calculated PAOI and RV/LV ratio revealed a significant correlation between them (r=0.36, p=0.023). D-dimer levels differed considerably among the mild (=40%), moderate (40%-60%) and severe (60%) groups constituted on the basis of PAOI (p=0.039). This difference stemmed from those in D-dimer levels in the mild group, PAOI =40 % and the severe group, PAOI 60% (p=0.02; Z= -2.328). In addition, D-dimer levels and PAOI revealed a positive correlation, but no similar correlation was determined between D-dimer levels and RV/LV. There were no significant correlations between IMA and D-dimer levels, PAOI and RV/LV ratios. In the biochemical determination of severity of PE based on radiological characteristics, D-dimer may be a more relevant marker than IMA, which has been proposed as a new marker.


Evidence review to investigate the support for subtypes of children with difficulty processing and integrating sensory information.
We investigated the evidence for subtypes in children with difficulty processing and integrating sensory information. Fifty-seven articles were incorporated into a systematic literature review; only 4 articles provided direct evidence for subtypes. These studies did not provide a comprehensive assessment of all sensory functions and sensory-based motor functions (i.e., praxis) and included different diagnostic groups. Therefore, generalized conclusions about subtypes could not be drawn. The other 53 studies reviewed provided meaningful information about strengths and challenges that children with difficulty processing and integrating sensory information demonstrate, but these studies were limited in scope. A principal theme was the importance of conducting comprehensive assessments of sensory-based functions, including multiple measures of sensory integrative functions such as praxis, sensory modulation, and sensory discrimination in children and adolescents with various clinical disorders. In addition, more consistency in the use of specific assessment tools will allow for synthesis of data across studies.


Wastewater retreatment and reuse system for agricultural irrigation in rural villages.
Climate changes and continuous population growth increase water demands that will not be met by traditional water resources, like surface and ground water. To handle increased water demand, treated municipal wastewater is offered to farmers for agricultural irrigation. This study aimed to enhance the effluent quality from worn-out sewage treatment facilities in rural villages, retreat effluent to meet water quality criteria for irrigation, and assess any health-related and environmental impacts from using retreated wastewater irrigation on crops and in soil. We developed the compact wastewater retreatment and reuse system (WRRS), equipped with filters, ultraviolet light, and bubble elements. A pilot greenhouse experiment was conducted to evaluate lettuce growth patterns and quantify the heavy metal concentration and pathogenic microorganisms on lettuce and in soil after irrigating with tap water, treated wastewater, and WRRS retreated wastewater. The purification performance of each WRRS component was also assessed. The study findings revealed that existing worn-out sewage treatment facilities in rural villages could meet the water quality criteria for treated effluent and also reuse retreated wastewater for crop growth and other miscellaneous agricultural purposes.


The combined effect of chemoprophylaxis with single dose rifampicin and immunoprophylaxis with BCG to prevent leprosy in contacts of newly diagnosed leprosy cases: a cluster randomized controlled trial (MALTALEP study).
Despite almost 30 years of effective chemotherapy with MDT, the global new case detection rate of leprosy has remained quite constant over the past years. New tools and methodologies are necessary to interrupt the transmission of M. leprae. Single-dose rifampicin (SDR) has been shown to prevent 57% of incident cases of leprosy in the first two years, when given to contacts of newly diagnosed cases. Immunization of contacts with BCG has been less well documented, but appears to have a preventive effect lasting up to 9 years. However, one major disadvantage is the occurrence of excess cases within the first year after immunization. The objective of this study is to examine the effect of chemoprophylaxis with SDR and immunoprophylaxis with BCG on the clinical outcome as well as on host immune responses and gene expression profiles in contacts of newly diagnosed leprosy patients. We hypothesize that the effects of both interventions may be complementary, causing the combined preventive outcome to be significant and long-lasting. Through a cluster randomized controlled trial we compare immunization with BCG alone with BCG plus SDR in contacts of new leprosy cases. Contact groups of around 15 persons will be established for each of the 1300 leprosy patients included in the trial, resulting in approximately 20,000 contacts in total. BCG will be administered to the intervention group followed by SDR, 2 months later. The control group will receive BCG only. In total 10,000 contacts will be included in both intervention arms over a 2-year period. Follow-up will take place one year as well as two years after intake. The primary outcome is the occurrence of clinical leprosy within two years. Simultaneously with vaccination and SDR, blood samples for in vitro analyses will be obtained from 300 contacts participating in the trial to determine the effect of these chemo- and immunoprophylactic interventions on immune and genetic host parameters. Combined chemoprophylaxis and immunoprophylaxis is potentially a very powerful and innovative tool aimed at contacts of leprosy patients that could reduce the transmission of M. leprae markedly. The trial intends to substantiate this potential preventive effect. Evaluation of immune and genetic biomarker profiles will allow identification of pathogenic versus (BCG-induced) protective host biomarkers and could lead to effective prophylactic interventions for leprosy using optimized tools for identification of individuals who are most at risk of developing disease. Netherlands Trial Register: NTR3087.


Growth phase-regulated expression of bolA and morphology of stationary-phase Escherichia coli cells are controlled by the novel sigma factor sigma S.
The novel sigma factor (sigma S) encoded by rpoS (katF) is required for induction of many growth phase-regulated genes and expression of a variety of stationary-phase phenotypes in Escherichia coli. Here we demonstrate that wild-type cells exhibit spherical morphology in stationary phase, whereas rpoS mutant cells remain rod shaped and are generally larger. Size reduction of E. coli cells along the growth curve is a continuous and at least biphasic process, the second phase of which is absent in rpoS-deficient cells and correlates with induction of the morphogene bolA in wild-type cells. Stationary-phase induction of bolA is dependent on sigma S. The "gearbox" a characteristic sequence motif present in the sigma S-dependent growth phase- and growth rate-regulated bolAp1 promoter, is not recognized by sigma S, since stationary-phase induction of the mcbA promoter, which also contains a gearbox, does not require sigma S, and other sigma S-controlled promoters do not contain gearboxes. However, good homology to the potential -35 and -10 consensus sequences for sigma S regulation is found in the bolAp1 promoter.


The Exaptation of HERV-H: Evolutionary Analyses Reveal the Genomic Features of Highly Transcribed Elements.
HERV-H endogenous retroviruses are thought to be essential to stem cell identity in humans. We embrace several decades of HERV-H research in order to relate the transcription of HERV-H loci to their genomic structure. We find that highly transcribed HERV-H loci are younger, more fragmented, and less likely to be present in other primate genomes. We also show that repeats in HERV-H LTRs are correlated to where loci are transcribed: type-I LTRs associate with stem cells while type-II repeats associate with embryonic cells. Our findings are generally in line with what is known about endogenous retrovirus biology but we find that the presence of the zinc finger motif containing region of gag is positively correlated with transcription. This leads us to suggest a possible explanation for why an unusually large proportion of HERV-H loci have been preserved in non-solo-LTR form.


Speed of onset, efficacy and tolerability of zolmitriptan nasal spray in the acute treatment of migraine: a randomised, double-blind, placebo-controlled study.
Migraine is a common, disabling condition that has a significant impact on patients and relatives, and is a considerable economic burden on society. Migraine patients want fast-acting treatments with high efficacy. Previous studies have demonstrated that orally administered formulations of zolmitriptan are rapidly and highly effective in the acute treatment of migraine. The objective of this study was to assess the efficacy, speed of onset and tolerability of the nasal spray formulation of zolmitriptan in migraine treatment. This multicentre, randomised, double-blind study recruited 2122 patients (aged 18-65 years) who had an established diagnosis of migraine (according to International Headache Society criteria), with or without aura. Patients were randomised to receive zolmitriptan 5mg nasal spray or placebo to treat up to two migraine attacks within 15 minutes of headache pain becoming moderate or severe. The primary endpoint was headache response (reduction in migraine pain from severe/moderate to mild/none) at 2 hours, 1 hour, 30 minutes and 15 minutes post-dose (analysed using a step-down approach). Secondary endpoints included headache response at 4 hours, pain-free rates at 30 minutes and 1, 2 and 4 hours, and sustained headache response and pain-free status at 24 hours post-dose. The headache response rate at 2 hours post-dose was 66.2% for the zolmitriptan group, compared with 35.0% for the placebo group (p < 0.001). Zolmitriptan nasal spray also produced significantly higher headache response rates than placebo at all earlier timepoints assessed, starting as early as 15 minutes post-dose (p < 0.001). Similar results were obtained for the analysis of the first attack. Significantly higher pain-free rates were obtained with zolmitriptan nasal spray, compared with placebo, from 15 minutes post-dose onward (p < 0.005). Zolmitriptan nasal spray was also significantly superior to placebo for headache response at 4 hours, sustained headache response at 24 hours and sustained pain-free rate at 24 hours. Zolmitriptan nasal spray was well tolerated, with most adverse events being of short duration and mild or moderate intensity. Zolmitriptan nasal spray is highly effective in the acute treatment of migraine and has a very fast onset of action, producing significant headache response and pain-free rates as early as 15 minutes post-dose (the earliest assessment in this study). In addition to the very fast onset of action, zolmitriptan nasal spray produced significantly higher sustained headache response and pain-free rates at 24 hours post-dose compared with placebo. These desirable efficacy outcomes were combined with good tolerability.


Prognostic significance of functional somatic symptoms in adolescence: a 15-year community-based follow-up study of adolescents with depression compared with healthy peers.
There is a lack of population-based long-term longitudinal research on mental health status and functional physical/somatic symptoms. Little is known about the long-term mental health outcomes associated with somatic symptoms or the temporal relationship between depression and such symptoms. This 15-year study followed up adolescents with depression and matched controls, screened from a population-based sample, who reported different numbers of somatic symptoms. The total population of 16-17-year-olds in Uppsala, Sweden, was screened for depression in 1991-1993. Adolescents who screened positive and an equal number of healthy controls took part in a semi-structured diagnostic interview. In addition, 21 different self-rated somatic symptoms were assessed. Sixty-four percent of those adolescents participated in a follow-up structured interview 15 years later. Somatic symptoms in adolescence predicted depression and other adult mental disorders regardless of the presence of adolescent depression. In adolescents with depression, the number of functional somatic symptoms predicted, in a dose response relationship, suicidal behavior, bipolar episodes, and psychotic episodes as well as chronic and recurrent depression. Contrary to expectations, the somatic symptoms of abdominal pain and perspiration without exertion better predicted depression than all DSM-IV depressive symptoms. Abdominal pain persisted as an independent strong predictor of depression and anxiety, even after controlling for other important confounders. Somatic symptoms in adolescence can predict severe adult mental health disorders. The number of somatic symptoms concurrent with adolescent depression is, in a stepwise manner, linked to suicidal attempts, bipolar disorders, psychotic disorders, and recurrent and chronic depression. These findings can be useful in developing treatment guidelines for patients with somatic symptoms.


Potential therapeutic properties of green tea polyphenols in Parkinson's disease.
Tea is one of the most frequently consumed beverages in the world. It is rich in polyphenols, a group of compounds that exhibit numerous biochemical activities. Green tea is not fermented and contains more catechins than black tea or oolong tea. Although clinical evidence is still limited, the circumstantial data from several recent studies suggest that green tea polyphenols may promote health and reduce disease occurrence, and possibly protect against Parkinson's disease and other neurodegenerative diseases. Green tea polyphenols have demonstrated neuroprotectant activity in cell cultures and animal models, such as the prevention of neurotoxin-induced cell injury. The biological properties of green tea polyphenols reported in the literature include antioxidant actions, free radical scavenging, iron-chelating properties, (3)H-dopamine and (3)H-methyl-4-phenylpyridine uptake inhibition, catechol-O-methyltransferase activity reduction, protein kinase C or extracellular signal-regulated kinases signal pathway activation, and cell survival/cell cycle gene modulation. All of these biological effects may benefit patients with Parkinson's disease. Despite numerous studies in recent years, the understanding of the biological activities and health benefits of green tea polyphenols is still very limited. Further in-depth studies are needed to investigate the safety and efficacy of green tea in humans and to determine the different mechanisms of green tea in neuroprotection.


Primary small cell carcinoma of the endometrium: report of a case with immunochemical studies.
Small cell carcinoma (SCC) is a well-known tumor that occurs predominantly in the lung. These tumors may also occur in the female genital tract, where it occurs most commonly in the cervix. Primary SCC of the endometrium is extremely rare. We report a case of an endometrial tumor that was a combination of an SCC and endometrioid adenocarcinoma with squamous components and that penetrated half of the thickness of the uterine wall. A 59-year-old, postmenopausal woman suffering from postmenopausal bleeding presented for care. The patient underwent a total abdominal hysterectomy, bilateral salpingo-oophorectomy, and pelvic and paraaortic lymph node dissection, followed by concurrent chemoradiotherapy. Immunohistochemical studies showed neuroendocrine differentiation with positive CD 56, chromogranin and synaptophysin markers. Immunohistochemical analyses are helpful in diagnosing and differentiating primary SCC of the endometrium from benign and malignant diseases of the endometrium.


Cyclosporine level at the second hour in pediatric hematopoietic stem cell transplant patients.
In this retrospective study, cyclosporine levels at the second hour (C2 levels) were measured during oral cyclosporine intake in 28 pediatric hematopoietic stem cell transplant patients, and the relations between cyclosporine dosage and C0, C2 levels, C2/C0 ratio, and cyclosporine-related adverse effects were investigated. Cyclosporine levels at the second hour levels were found to be significantly lower in children younger than 7 years old, suggesting age-related differences in absorption and metabolism of the drug. There were statistically significant correlations of both C0 and C2 levels with blood creatinine values. In addition, a statistically significant negative relation was found between C0 and C2 levels and serum potassium levels; this unexpected finding was attributed to multiple drug effects in the early posttransplant period. The common adverse effects of cyclosporine (gingival overgrowth, gynecomastia, and hypertrichosis) were also evaluated in this study, and no correlation was found between those adverse effects and C0, C2 levels, C2/C0 ratio, and cyclosporine dosage. In the present study, despite the highly significant correlation of C2 levels with renal and metabolic effects, in pediatric hematopoietic stem cell transplant patients, measurement of C2 levels as a standard practice did not provide an advantage over C0 monitoring. However, the preliminary results suggest that C2 level monitoring could be useful in selected patients with increased risk of renal toxicity or in states where a better estimation of gastrointestinal absorption is needed.


Sleep stability with home sleep recording and automatic sleep stage analysis.
Three home sleep recordings were conducted in 12 normal subjects using the Oxford Medilog 9000 system with automatic sleep stage analysis (SS90III). Each night was separated by 1 week. No significant differences were noted for any sleep parameter, with the mean intraindividual variation less than 20% for most parameters. This indicates that sleep patterns are stable over time.


Cross-cultural differences in hemisphericity: EEG asymmetry discriminates between Japanese and Westerners.
This study was designed to test the hypothesis that Japanese subjects exhibit different patterns of resting EEG asymmetry compared with Westerners. EEG was recorded from the left and right temporal and parietal scalp regions in bilingual Japanese and Western subjects during eyes-open and eyes-closed rest periods before and after the performance of a series of cognitive tasks. Alpha activity was integrated and digitized. Japanese subjects were found to exhibit greater relative right-sided parietal activation during the eyes closed condition. This difference was found to be a function of greater left hemisphere activation among the Westerners. Various possible contributors to this cross-cultural differences are discussed.


A Dirichlet process model for classifying and forecasting epidemic curves.
A forecast can be defined as an endeavor to quantitatively estimate a future event or probabilities assigned to a future occurrence. Forecasting stochastic processes such as epidemics is challenging since there are several biological, behavioral, and environmental factors that influence the number of cases observed at each point during an epidemic. However, accurate forecasts of epidemics would impact timely and effective implementation of public health interventions. In this study, we introduce a Dirichlet process (DP) model for classifying and forecasting influenza epidemic curves. The DP model is a nonparametric Bayesian approach that enables the matching of current influenza activity to simulated and historical patterns, identifies epidemic curves different from those observed in the past and enables prediction of the expected epidemic peak time. The method was validated using simulated influenza epidemics from an individual-based model and the accuracy was compared to that of the tree-based classification technique, Random Forest (RF), which has been shown to achieve high accuracy in the early prediction of epidemic curves using a classification approach. We also applied the method to forecasting influenza outbreaks in the United States from 1997-2013 using influenza-like illness (ILI) data from the Centers for Disease Control and Prevention (CDC). We made the following observations. First, the DP model performed as well as RF in identifying several of the simulated epidemics. Second, the DP model correctly forecasted the peak time several days in advance for most of the simulated epidemics. Third, the accuracy of identifying epidemics different from those already observed improved with additional data, as expected. Fourth, both methods correctly classified epidemics with higher reproduction numbers (R) with a higher accuracy compared to epidemics with lower R values. Lastly, in the classification of seasonal influenza epidemics based on ILI data from the CDC, the methods' performance was comparable. Although RF requires less computational time compared to the DP model, the algorithm is fully supervised implying that epidemic curves different from those previously observed will always be misclassified. In contrast, the DP model can be unsupervised, semi-supervised or fully supervised. Since both methods have their relative merits, an approach that uses both RF and the DP model could be beneficial.


Mapping cortical change across the human life span.
We used magnetic resonance imaging and cortical matching algorithms to map gray matter density (GMD) in 176 normal individuals ranging in age from 7 to 87 years. We found a significant, nonlinear decline in GMD with age, which was most rapid between 7 and about 60 years, over dorsal frontal and parietal association cortices on both the lateral and interhemispheric surfaces. Age effects were inverted in the left posterior temporal region, where GMD gain continued up to age 30 and then rapidly declined. The trajectory of maturational and aging effects varied considerably over the cortex. Visual, auditory and limbic cortices, which are known to myelinate early, showed a more linear pattern of aging than the frontal and parietal neocortices, which continue myelination into adulthood. Our findings also indicate that the posterior temporal cortices, primarily in the left hemisphere, which typically support language functions, have a more protracted course of maturation than any other cortical region.


Follow-Up Care for Breast and Colorectal Cancer Across the Globe: Survey Findings From 27 Countries.
The purpose of this study was to describe follow-up care for breast and colorectal cancer survivors in countries with varying levels of resources and highlight challenges regarding posttreatment survivorship care. We surveyed one key stakeholder from each of 27 countries with expertise in survivorship care on questions including the components/structure of follow-up care, delivery of treatment summaries and survivorship care plans, and involvement of primary care in survivorship. Descriptive analyses were performed to characterize results across countries and variations between the WHO income categories (low, middle, high). We also performed a qualitative content analysis of narratives related to survivorship care challenges to identify major themes. Seven low- or /lower-middle-income countries (LIC/LMIC), seven upper-middle-income countries (UMIC), and 13 high-income countries (HICs) were included in this study. Results indicate that 44.4% of countries with a National Cancer Control Plan currently address survivorship care. Additional findings indicate that HICs use guidelines more often than those in LICs/LMICs and UMICs. There was great variation among countries regardless of income level. Common challenges include issues with workforce, communication and care coordination, distance/transportation issues, psychosocial support, and lack of focus on follow-up care. This information can guide researchers, providers, and policy makers in efforts to improve the quality of survivorship care on a national and global basis. As the number of cancer survivors increases globally, countries will need to prioritize their long-term needs. Future efforts should focus on efforts to bridge oncology and primary care, building international partnerships, and implementation of guidelines.


Reduced representation approaches to interrogate genome diversity in large repetitive plant genomes.
Technology and software improvements in the last decade now provide methodologies to access the genome sequence of not only a single accession, but also multiple accessions of plant species. This provides a means to interrogate species diversity at the genome level. Ample diversity among accessions in a collection of species can be found, including single-nucleotide polymorphisms, insertions and deletions, copy number variation and presence/absence variation. For species with small, non-repetitive rich genomes, re-sequencing of query accessions is robust, highly informative, and economically feasible. However, for species with moderate to large sized repetitive-rich genomes, technical and economic barriers prevent en masse genome re-sequencing of accessions. Multiple approaches to access a focused subset of loci in species with larger genomes have been developed, including reduced representation sequencing, exome capture and transcriptome sequencing. Collectively, these approaches have enabled interrogation of diversity on a genome scale for large plant genomes, including crop species important to worldwide food security.


Interpolation revisited.
Based on the theory of approximation, this paper presents a unified analysis of interpolation and resampling techniques. An important issue is the choice of adequate basis functions. We show that, contrary to the common belief, those that perform best are not interpolating. By opposition to traditional interpolation, we call their use generalized interpolation; they involve a prefiltering step when correctly applied. We explain why the approximation order inherent in any basis function is important to limit interpolation artifacts. The decomposition theorem states that any basis function endowed with approximation order can be expressed as the convolution of a B-spline of the same order with another function that has none. This motivates the use of splines and spline-based functions as a tunable way to keep artifacts in check without any significant cost penalty. We discuss implementation and performance issues, and we provide experimental evidence to support our claims.


Mammalian chymotrypsin-like enzymes. Comparative reactivities of rat mast cell proteases, human and dog skin chymases, and human cathepsin G with peptide 4-nitroanilide substrates and with peptide chloromethyl ketone and sulfonyl fluoride inhibitors.
The extended substrate binding sites of several chymotrypsin-like serine proteases, including rat mast cell proteases I and II (RMCP I and II, respectively) and human and dog skin chymases, have been investigated by using peptide 4-nitroanilide substrates. In general, these enzymes preferred a P1 Phe residue and hydrophobic amino acid residues in P2 and P3. A P2 Pro residue was also found to be quite acceptable. The S4 subsites of these enzymes are less restrictive than the other subsites investigated. The substrate specificity of these enzymes was also investigated by using substrates which contain model desmosine residues and peptides with amino acid sequences of the physiologically important substrates angiotensin I and angiotensinogen and alpha 1-antichymotrypsin, the major plasma inhibitor for chymotrypsin-like enzymes. These substrates were less reactive than the most reactive tripeptide reported here, Suc-Val-Pro-Phe-NA. The thiobenzyl ester Suc-Val-Pro-Phe-SBzl was found to be an extremely reactive substrate for the enzymes tested and was 6-171-fold more reactive than the 4-nitroanilide substrate. The four chymotrypsin-like enzymes were inhibited by chymostatin and N-substituted saccharin derivatives which had KI values in the micromolar range. In addition, several potent peptide chloromethyl ketone and substituted benzenesulfonyl fluoride irreversible inhibitors for these enzymes were discovered. The most potent sulfonyl fluoride inhibitor for RMCP I, RMCP II, and human skin chymase, 2-(Z-NHCH2CONH)C6H4SO2F, had kobsd/[I] values of 2500, 270, and 1800 M-1 s-1, respectively. The substrates and inhibitors reported here should be extremely useful in elucidating the physiological roles of these proteases.


A comparison of the performance of quality controls prepared from spiked, fortified and authentic hair for ethyl glucuronide analysis.
Ethyl glucuronide (EtG) quantification in hair was assessed using quality controls prepared by three methods: (a) spiking hair samples with known concentrations of EtG, (b) fortifying hair by incubation of blank hair with EtG for several days or (c) use of authentic hair samples positive for EtG. A liquid chromatography tandem mass spectrometry (LC-MS/MS) method was developed on a Shimadzu model 8030 instrument and validated for the quantification of EtG. For two concentration levels, approximately 50 and 500 pg/mg QCs, EtG concentrations were measured in duplicate (N=2) on 8 days (N=16) and intra-assay precision (repeatability) and inter-assay precision determined using one-way analysis of variance. EtG concentrations measured in authentic hair exhibited poor intra-assay precision, with coefficients of variation of 25.1 and 20.9%, compared with 17.7 and 18.5% for fortified hair and 17.4 and 11.3% for spiked hair, for the lower and higher concentrations respectively. The inter-assay precision for authentic hair was also poorer, 35.7 and 22.5%, compared with fortified (28.2 and 19.8%) and spiked (18.4 and 13.2%) hair for the lower and higher concentrations. Although spiked QCs resulted in a better repeatability and inter-assay precision, the values obtained for QCs prepared from fortified and authentic hair are likely to be more representative of case specimens. These results have implications on the interpretation of EtG concentrations when spiked QCs are used to validate methods.


Factors determining symptoms in heart failure: comparison of fast and slow exercise tests.
Factors determining the symptoms of breathlessness and fatigue in patients with congestive heart failure were investigated by comparing the response to slow and fast exercise. Symptom limited oxygen consumption (maximal); minute ventilation, mean pulmonary capillary wedge pressure; and arterial blood gases, pH, and lactate concentrations were measured during treadmill exercise using a slow protocol in 25 men (age 34-67 years) with congestive heart failure (New York Heart Association class II-III). Ten of these patients were also exercised according to a rapid protocol. Exercise was terminated by fatigue in 23/25 patients after the slow test and by breathlessness in all patients after the rapid test. Exercise capacity (maximal oxygen consumption and exercise duration) was not related to resting or exercise pulmonary capillary wedge pressure or the change in pulmonary capillary wedge pressure during exercise, nor was there any difference in pulmonary capillary wedge pressure at the end of exercise within individuals between the fast and slow tests. Minute ventilation was greater (51 vs 43 1/min), peak exercise lactate concentration higher (3.7 vs 2.2 mmol/l), and the change in pH from the resting state was greater (0.06 vs 0.02) during the rapid test than during the slow test. The sensation of breathlessness in congestive heart failure is not simply related to raised pulmonary capillary wedge pressure, but may in part be due to stimulation of peripheral chemoreceptors in response to metabolic acidosis.


C-terminal BRE overexpression in 11q23-rearranged and t(8;16) acute myeloid leukemia is caused by intragenic transcription initiation.
Overexpression of the BRE (brain and reproductive organ-expressed) gene defines a distinct pediatric and adult acute myeloid leukemia (AML) subgroup. Here we identify a promoter enriched for active chromatin marks in BRE intron 4 causing strong biallelic expression of a previously unknown C-terminal BRE transcript. This transcript starts with BRE intron 4 sequences spliced to exon 5 and downstream sequences, and if translated might code for an N terminally truncated BRE protein. Remarkably, the new BRE transcript was highly expressed in over 50% of 11q23/KMT2A (lysine methyl transferase 2A)-rearranged and t(8;16)/KAT6A-CREBBP cases, while it was virtually absent from other AML subsets and normal tissues. In gene reporter assays, the leukemia-specific fusion protein KMT2A-MLLT3 transactivated the intragenic BRE promoter. Further epigenome analyses revealed 97 additional intragenic promoter marks frequently bound by KMT2A in AML with C-terminal BRE expression. The corresponding genes may be part of a context-dependent KMT2A-MLLT3-driven oncogenic program, because they were higher expressed in this AML subtype compared with other groups. C-terminal BRE might be an important contributor to this program because in a case with relapsed AML, we observed an ins(11;2) fusing CHORDC1 to BRE at the region where intragenic transcription starts in KMT2A-rearranged and KAT6A-CREBBP AML.


Modeling buprenorphine reduction of fentanyl-induced respiratory depression.
BACKGROUNDPotent synthetic opioids, such as fentanyl, are increasingly abused, resulting in unprecedented numbers of fatalities from respiratory depression. Treatment with the high-affinity mu-opioid receptor partial agonist buprenorphine may prevent fatalities by reducing binding of potent opioids to the opioid receptor, limiting respiratory depression.METHODSTo characterize buprenorphine-fentanyl interaction at the level of the mu-opioid receptor in 2 populations (opioid-naive individuals and individuals who chronically use high-dose opioids), the effects of escalating i.v. fentanyl doses with range 0.075-0.35 mg/70 kg (opioid naive) and 0.25-0.70 mg/70 kg (chronic opioid use) on iso-hypercapnic ventilation at 2-3 background doses of buprenorphine (target plasma concentrations range: 0.2-5 ng/mL) were quantified using receptor association/dissociation models combined with biophase distribution models.RESULTSBuprenorphine produced mild respiratory depression, while high doses of fentanyl caused pronounced respiratory depression and apnea in both populations. When combined with fentanyl, buprenorphine produced a receptor binding-dependent reduction of fentanyl-induced respiratory depression in both populations. In individuals with chronic opioid use, at buprenorphine plasma concentrations of 2 ng/mL or higher, a protective effect against high-dose fentanyl was observed.CONCLUSIONOverall, the results indicate that when buprenorphine mu-opioid receptor occupancy is sufficiently high, fentanyl is unable to activate the mu-opioid receptor and consequently will not cause further respiratory depression in addition to the mild respiratory effects of buprenorphine.TRIAL REGISTRATIONTrialregister.nl, no. NL7028 (https://www.trialregister.nl/trial/7028)FUNDINGIndivior Inc., North Chesterfield, Virginia, USA.


Rational design of universal immunotherapy for TfR1-tropic arenaviruses.
Certain arenaviruses that circulate in rodent populations can cause life-threatening hemorrhagic fevers when they infect humans. Due to their efficient transmission, arenaviruses pose a severe risk for outbreaks and might be exploited as biological weapons. Effective countermeasures against these viruses are highly desired. Ideally, a single remedy would be effective against many or even all the pathogenic viruses in this family. However, despite the fact that all pathogenic arenaviruses from South America utilize transferrin receptor 1 (TfR1) as a cellular receptor, their viral glycoproteins are highly diversified, impeding efforts to isolate cross-neutralizing antibodies. Here we address this problem using a rational design approach to target TfR1-tropic arenaviruses with high potency and breadth. The pan-reactive molecule is highly effective against all arenaviruses that were tested, offering a universal therapeutic approach. Our design scheme avoids the shortcomings of previous immunoadhesins and can be used to combat other zoonotic pathogens.


Substrate-assisted mechanism of catalytic hydrolysis of misaminoacylated tRNA required for protein synthesis fidelity.
d-aminoacyl-tRNA-deacylase (DTD) prevents the incorporation of d-amino acids into proteins during translation by hydrolyzing the ester bond between mistakenly attached amino acids and tRNAs. Despite extensive study of this proofreading enzyme, the precise catalytic mechanism remains unknown. Here, a combination of biochemical and computational investigations has enabled the discovery of a new substrate-assisted mechanism of d-Tyr-tRNATyr hydrolysis by Thermus thermophilus DTD. Several functional elements of the substrate, misacylated tRNA, participate in the catalysis. During the hydrolytic reaction, the 2'-OH group of the А76 residue of d-Tyr-tRNATyr forms a hydrogen bond with a carbonyl group of the tyrosine residue, stabilizing the transition-state intermediate. Two water molecules participate in this reaction, attacking and assisting ones, resulting in a significant decrease in the activation energy of the rate-limiting step. The amino group of the d-Tyr aminoacyl moiety is unprotonated and serves as a general base, abstracting the proton from the assisting water molecule and forming a more nucleophilic ester-attacking species. Quantum chemical methodology was used to investigate the mechanism of hydrolysis. The DFT-calculated deacylation reaction is in full agreement with the experimental data. The Gibbs activation energies for the first and second steps were 10.52 and 1.05 kcal/mol, respectively, highlighting that the first step of the hydrolysis process is the rate-limiting step. Several amino acid residues of the enzyme participate in the coordination of the substrate and water molecules. Thus, the present work provides new insights into the proofreading details of misacylated tRNAs and can be extended to other systems important for translation fidelity.


Cytogenic investigations in the assessment of response to treatment in neuroblastoma.
In a patient with stage IV disseminated neuroblastoma treated by chemotherapy extensive cytogenetic investigations were performed on the residual primary tumour and bone marrow immediately before myeloablative treatment and autologous marrow rescue. Two abnormal clones both showing lp+, a characteristic abnormality of neuroblastoma, were detected in cells from the residual primary tumour, providing direct evidence of persisting viable tumour. Such investigations should be a routine part of the assessment of response to treatment in patients with neuroblastoma, and could be extended to others in whom "second look" surgery is performed.


Survey of practices around pharmaceutical company funding for continuing professional development among medical oncologists and trainees in Australia.
The completion of continuing professional development (CPD) is mandatory for medical oncologists and trainees (MO&T). Pharmaceutical companies may fund some CPD activities, but there is increasing debate about the potential for conflicts of interest (COI). To assess current practices around funding to attend CPD activities. An electronic survey was distributed to Australian MO&T. The survey asked questions about current practices, institutional policies and perceptions about attending CPD funded by pharmaceutical companies. The design looked at comparing responses between MO&T as well as their understanding of and training around institutional and ethical process. A total of 157 of 653 (24%) responses was received, the majority from MO (76%). Most CPD activities attended by MO&T were self-funded (53%), followed by funding from institutions (19%), pharmaceutical companies (16%) and salary award (16%). Most institutions allowed MO&T to receive CPD funding from professional organisations (104/157, 66%) or pharmaceutical companies (90/157, 57%). A minority of respondents (13/157, 8%) reported that the process to use pharmaceutical funds had been considered by an ethics committee. Although 103/157 (66%) had received pharmaceutical funding for CPD, most (109/157, 69%) reported never receiving training about potential COI. The lack of education was more noticeable among trainees (odds ratio (OR) 8.61, P = 0.02). MO&T acknowledged the potential bias towards a pharmaceutical product (P = 0.05) but believed there was adequate separation between themselves and pharmaceutical companies (P < 0.01). Majority of CPD attended by MO&T is self-funded. There is lack of clarity in institutional policies regarding external funding support for CPD activities. Formal education about potential COI is lacking.


A gene therapeutic approach to correct splice defects with modified U1 and U6 snRNPs.
Splicing is an essential cellular process to generate mature transcripts from pre-mRNA. It requires the splice factor U1 small nuclear ribonucleoprotein (U1), which promotes exon recognition by base-pairing interaction with the splice donor site (SD). After U1 dissociation, exon recognition is maintained by U6 small nuclear ribonucleoproteins (U6). It has been shown that SD mutations lower the binding affinity of U1 and cause splice defects in about 10% of patients with monogenetic diseases. U1 isoforms specifically designed to bind the mutated SD with increased affinity can correct these splice defects. We investigated the applicability of this gene therapeutic approach for different mutated SD positions. A minigene-based splicing assay was established to study a typical SD derived from the gene BBS1. We found that mutations at seven SD positions caused splice defects. In four cases, mutation-adapted U1 isoforms completely corrected these splice defects. Partial correction was found for splice defects induced by the mutation at SD position +5. The limited therapeutic efficacy at this position was alleviated by applying a combined treatment with mutation-adapted U1 and U6. The sequence complementarity between U6 and three SD positions (+4, +5,and +6) was relevant for the outcome of the therapy. Between 30 and 100% of the normal transcripts can be restored. The treatment significantly decreased both exon skipping and intron retention. Massive missplicing of off-target transcripts was not detected. Our study helps to assess the therapeutic efficacy of mutation-adapted U snRNAs in gene therapy and illustrates their strong potential to correct splice defects, which cause many different inherited conditions.


Splenectomy and the risk of sepsis: a population-based cohort study.
We sought to estimate the long-term risk of sepsis in patients who underwent splenectomy before, during, and after implementation of vaccination. Because patients who have undergone splenectomy are considered at increased risk of bacterial sepsis, they typically receive vaccination, education, and occasionally antibiotic prophylaxis. However, the extent to which these interventions have actually reduced the risk of sepsis remains unclear. Retrospective cohort study encompassing all patients in the Swedish national inpatient register, who underwent splenectomy in 1970-2009. Patients were followed for hospitalization for or death from sepsis, as identified using national inpatient and cause of death registers. Relative risks, comparing patients to the background population were expressed as standardized incidence ratios (SIRs) and standardized mortality ratios (SMRs). Altogether, 20,132 splenectomized patients were included. The overall SIR for hospitalization for sepsis was 5.7 [95% confidence interval (CI), 5.6-6.0]. However, risks depended on the indication for splenectomy, with SIRs varying from 3.4 (95% CI, 3.0-3.8) for trauma patients to 18 (95% CI, 16-19) for patients with hematologic malignancies. SMRs ranged from 3.1 (95% CI, 2.1-4.3) for trauma to 8.7 (95% CI, 6.8-11) for hematologic disease. In regression analyses adjusting for age at splenectomy, follow-up time, sex, and calendar year of splenectomy, there were no significant risk decreases after implementation of routine vaccination, except for in patients with malignant and non-malignant hematologic disease. The risk of hospitalization or death from sepsis is high in patients who previously underwent splenectomy and depends on the indication for splenectomy. The effectiveness of current vaccination practices warrants further evaluation.


Reproducibility and clinical evaluation of rebound tonometry.
To establish the reproducibility of a rebound tonometer in humans and the effect of corneal thickness on measurements, comparing it with Goldmann applanation tonometer. In a first study designed to examine the reliability of the RBT, three experienced ophthalmologists undertook three consecutive intraocular pressure (IOP) measurements in 12 eyes of 12 normal subjects. A cross-sectional study was then performed to compare measurements obtained using the two tonometers in 147 eyes of 85 patients with ocular hypertension or glaucoma. Intraobserver coefficients of correlation obtained in the reproducibility study were 0.82, 0.73, and 0.87. Interobserver correlation was 0.82. There was a good correlation between IOP readings obtained by the RBT and the GAT (r = 0.865, P < 0.0001). RBT readings were consistently higher than GAT measurements (median difference, 1.8 +/- 2.8 mm Hg). A Bland-Altman plot indicated the 95% limits of agreement between the two methods were -3.7 to 7.3 mm Hg (slope = -0.022, P = 0.618). Using RBT, the point that best discriminated between patients with an IOP < or = 21 mm Hg and those with >21 mm Hg, as determined by the GAT was >23 mm Hg (sensitivity, 70.5%; specificity, 95.1%). In terms of pachymetry, the two tonometers behaved in a similar way, with correlation observed between IOP measurements and central corneal thickness. Rebound tonometry is a reproducible method of determining IOP in humans. In general, it tends to overestimate IOP compared with Goldmann applanation tonometry. The tonometers used in both methods are similarly affected by pachymetry.


Developing a standard short questionnaire for the assessment of patient satisfaction with out-of-hours primary care.
Governmental reviews of out-of-hours services in England and Scotland have recommended that a standard questionnaire should be used to assess patient satisfaction. This is important because of the rapid introduction of new forms of care. To produce a brief, reliable and valid measure of patient satisfaction for use by a wide variety of providers of out-of-hours primary care. The Short Questionnaire for Out-of-Hours care was designed and compared with a longer questionnaire which had been validated and used in earlier research. Questionnaires were sent to 1906 people contacting an out-of-hours GP co-operative. Three versions of the short questionnaire were used with different formats. Analysis compared the response rates, measurement properties, concurrent and construct validity of the short and long questionnaires, and of different versions of the short questionnaire. There was no significant difference in the overall response rates obtained from the short or long questionnaires (45.7% versus 41.9%; P = 0.17). The effective response rate of questionnaires from which all satisfaction scales could be calculated was higher for the short questionnaire (43.0% versus 36.4%; P = 0.01). There were no significant differences in response rates or distribution of responses between different versions of the short questionnaire. There was moderate agreement between items on the short questionnaire and corresponding scales on the long questionnaire. Scores using the short questionnaire showed anticipated relationships with the age and sex of patients and with characteristics of how the service was delivered. The SQOC is valid and reliable for routine service use.


Evaluating Internet-Based Information on Physical Activity for Children and Youth With Physical Disabilities.
Parents are an important source of support for facilitating physical activity in children and youth with disabilities (CYWD). Approximately 70% of parents report using the Internet to search for information regarding their children's health. This study examined the theoretical content of physical activity information contained on publicly available Web sites targeting parents of CYWD. Web sites were amassed using Google, a combination of various search terms, and predetermined inclusion criteria. The Web sites were coded and analyzed using the content-analysis approach to the theory of specified persuasive educational communication. Half of the total Web site content targeted knowledge-based information and messages concerning outcome expectancies. Web sites infrequently included messages concerning self-regulation. Furthermore, the majority of the Web sites were accumulated using the generic term disability. This research highlights the gaps between theory and practice, emphasizing the need for better knowledge-translation practices.


Effect of the Tokyo 2020 Summer Olympic Games on COVID-19 incidence in Japan: a synthetic control approach.
The Tokyo 2020 Summer Olympic Games (23 July-8 August 2021) were held in the middle of Japan's fifth wave of COVID-19, when the number of cases was on the rise, and coincided with the fourth state of emergency implemented by the host city, Tokyo. This study aimed to assess whether the hosting of the Games was associated with a change in the number of COVID-19 cases in Japan using a synthetic control method. A weighted average of control countries with a variety of predictors was used to estimate the counterfactual trajectory of daily COVID-19 cases per 1 000 000 population in the absence of the Games in Japan. Outcome and predictor data were extracted using official and open sources spanning several countries. The predictors comprise the most recent country-level annual or daily data accessible during the Games, including the stringency of the government's COVID-19 response, testing capacity and vaccination capacity; human mobility index; electoral democracy index and demographic, socioeconomic, health and weather information. After excluding countries with missing data, 42 countries were ultimately used as control countries. The number of observed cases per 1 000 000 population on the last day of the Games was 109.2 (7-day average), which was 115.7% higher than the counterfactual trajectory comprising 51.0 confirmed cases per 1 000 000 population. During the Olympic period (since 23 July), the observed cumulative number of cases was 61.0% higher than the counterfactual trajectory, comprising 143 072 and 89 210 confirmed cases (p=0.023), respectively. The counterfactual trajectory lagged 10 days behind the observed trends. Given the increasing likelihood that new emerging infectious diseases will be reported in the future, we believe that the results of this study should serve as a sentinel warning for upcoming mega-events during COVID-19 and future pandemics.


The genomic and transcriptomic landscape of a HeLa cell line.
HeLa is the most widely used model cell line for studying human cellular and molecular biology. To date, no genomic reference for this cell line has been released, and experiments have relied on the human reference genome. Effective design and interpretation of molecular genetic studies performed using HeLa cells require accurate genomic information. Here we present a detailed genomic and transcriptomic characterization of a HeLa cell line. We performed DNA and RNA sequencing of a HeLa Kyoto cell line and analyzed its mutational portfolio and gene expression profile. Segmentation of the genome according to copy number revealed a remarkably high level of aneuploidy and numerous large structural variants at unprecedented resolution. Some of the extensive genomic rearrangements are indicative of catastrophic chromosome shattering, known as chromothripsis. Our analysis of the HeLa gene expression profile revealed that several pathways, including cell cycle and DNA repair, exhibit significantly different expression patterns from those in normal human tissues. Our results provide the first detailed account of genomic variants in the HeLa genome, yielding insight into their impact on gene expression and cellular function as well as their origins. This study underscores the importance of accounting for the strikingly aberrant characteristics of HeLa cells when designing and interpreting experiments, and has implications for the use of HeLa as a model of human biology.


Binding of hexabrachion (tenascin) to the extracellular matrix and substratum and its effect on cell adhesion.
Hexabrachion is a large glycoprotein of the extracellular matrix (ECM) that is prominent in embryogenesis, wound healing and tumorigenesis. Because of the role of extracellular matrix proteins in the regulation of cell differentiation and migration, the interaction of hexabrachion with cells as well as with other components of the ECM is of great interest. Early reports suggested that hexabrachion does not bind to fibronectin or gelatin but does bind to chondroitin sulfate proteoglycans. However, more recent reports have suggested that hexabrachion binds to fibronectin and inhibits cell adhesion as well as cell migration on fibronectin. We have found no evidence of strong hexabrachion-fibronectin binding on either a solid-phase ELISA assay or in a fluid-phase sedimentation assay in which the reactants were allowed to dissociate. However, hexabrachion sedimentation was accelerated in a gradient containing fibronectin throughout. This demonstrates an association between hexabrachions and fibronectin, but the complex is apparently weak and readily reversible. The solid-phase ELISA also shows no evidence of hexabrachion binding to gelatin, laminin or types I, III, IV or V collagen. Hexabrachion does not support strong cell attachment of the cell lines tested. Moreover, hexabrachion can inhibit cell attachment to fibronectin. We demonstrate here that this inhibition requires the hexabrachion to be able to bind to the plastic substratum. The results suggest that hexabrachion inhibition is via a steric inhibition. When the hexabrachion molecules bind to the plastic, they cover up a significant fraction of the underlying fibronectin molecules. Antibody studies are presented that show that hexabrachion can nonspecifically block access of immunoglobulin G molecules to the underlying matrix. This steric blocking is not unique to hexabrachion.


Utilization and determinants of adjuvant therapy among older patients who receive curative surgery for pancreatic cancer.
We conducted a population-based study to describe the utilization, determinants, and survival effects of adjuvant therapies after surgery among older patients with pancreatic cancer. Using Surveillance, Epidemiology, and End Results-Medicare data, we identified patients older than 65 years who received surgical resection for pancreatic cancer during 1992-2002. We constructed multiple logistic regression models to examine patient, clinical, and hospital factors associated with receiving adjuvant therapy. Cox proportional hazards models were used to examine the effect of therapy on survival. Approximately 49% of patients received adjuvant therapy after surgery. Patient factors associated with increased receipt of adjuvant therapy included more recent diagnosis, younger age, stage II disease, higher income, and geographic location. Hospital factors associated with increased receipt of adjuvant therapy included cooperative group membership and larger size. Adjuvant treatments associated with a significant reduction in 2-year mortality (relative to surgery alone) were chemoradiation or radiation alone but not chemotherapy alone. Our findings suggest that adjuvant chemoradiation and, to a lesser degree, radiation only are associated with a reduction in the risk of mortality among older patients who undergo surgery for pancreatic cancer. However, receipt of adjuvant therapy varied by period and geography as well as by certain patient and hospital factors.


Supramolecular architecture of severe acute respiratory syndrome coronavirus revealed by electron cryomicroscopy.
Coronavirus particles are enveloped and pleomorphic and are thus refractory to crystallization and symmetry-assisted reconstruction. A novel methodology of single-particle image analysis was applied to selected virus features to obtain a detailed model of the oligomeric state and spatial relationships among viral structural proteins. Two-dimensional images of the S, M, and N structural proteins of severe acute respiratory syndrome coronavirus and two other coronaviruses were refined to a resolution of approximately 4 nm. Proteins near the viral membrane were arranged in overlapping lattices surrounding a disordered core. Trimeric glycoprotein spikes were in register with four underlying ribonucleoprotein densities. However, the spikes were dispensable for ribonucleoprotein lattice formation. The ribonucleoprotein particles displayed coiled shapes when released from the viral membrane. Our results contribute to the understanding of the assembly pathway used by coronaviruses and other pleomorphic viruses and provide the first detailed view of coronavirus ultrastructure.


Fretting corrosion behaviour of Ti-6Al-4V reinforced with zirconia in foetal bovine serum.
Fretting corrosion is a critical challenge in the design of hip prosthesis used in total hip arthroplasty (THA) surgeries. Currently, the design of hip implants includes a tapered junction which introduces additional interfaces that connect different parts of the hip implant such as the femoral neck and head or stem and neck interface. Micro motions that occur under the influence of load, together with chemical changes in the host environment, make these interfaces susceptible to tribocorrosion processes, particularly fretting corrosion. Commonly used metallic biomaterials are based on stainless steels, cobalt chrome-based alloys as well as titanium and titanium alloys. Each of these materials possess some degree of limitations, particularly where tribocorrosion events are concerned. Titanium alloy Ti-6Al-4V is widely used in biomedical applications for non-bearing components of total joint arthroplasty (TJA) surgeries. Its poor wear resistance continues to remain a challenge in load-bearing joints where parts articulate against one another as in the case of modular junctions. Some of the attempts made to improve the wear properties of Ti-6Al-4V is through the incorporation of second phase particles like ceramics in its matrix to produce metal matrix composites of Ti-6Al-4V. The aim of this work is to investigate the effect of zirconia reinforcement on spark plasma sintered Ti-6Al-4V composites (zirconium oxide particles incorporated into Ti-6Al-4V matrix) on the fretting corrosion properties of Ti-6Al-4V. Fretting corrosion tests were carried out on as-sintered Ti-6Al-4V and Ti-6Al-4V with 5 and 10 wt.% ZrO2. The tests were carried out in foetal bovine serum under applied normal loads of 85 and 115 N using the cylinder-on-flat contact configuration. The evolution of OCP, dissipated energy and friction coefficient were recorded throughout the test. Microstructural analysis of the samples before fretting corrosion tests showed the presence of globular agglomerates throughout the Ti-6Al-4V matrix due to zirconia additions; the volume of the agglomerates was higher in the composites having 10 wt.% ZrO2. Ti-6Al-4V composites having zirconia additions produced a nobler OCP during fretting in foetal bovine serum, compared to pure Ti-6Al-4V. Furthermore, the fretting corrosion results showed a significant improvement in the tribocorrosion resistance of Ti-6Al-4V with 10 wt.% ZrO2 at all loads. This composition also produced the least amount of degradation. and metal ion release. Mechanical data showed that increasing the applied normal load promoted a transition from gross slip to partial slip conditions for all compositions. Partial slip was found to be prevalent at a higher normal load (drastic decrease of the dissipated energy and consequently the friction coefficient). This mechanical condition prevents a large amount of degradation.


The effects of spinal manipulation on the intensity of emotional arousal in phobic subjects exposed to a threat stimulus: a randomized, controlled, double-blind clinical trial.
To determine the effect of spinal manipulation upon the intensity of emotional arousal in phobic subjects exposed to a threat stimulus. Randomized, controlled, double-blind clinical trial. Community college campus. Eighteen phobic community college student volunteers randomized into treatment and control groups. Visual analog scale (VAS) and pulse rates were obtained in response to the subjects' viewing their phobogenic stimulus. Spinal manipulation was performed while the subjects experienced emotional responses. Manual muscle testing was utilized to ascertain the associated spinal segments and involved emotion. Data were analyzed using analysis of variance for a repeated measures experimental design and Least Significant Differences (LSDs) for mean comparisons. Baseline, preintervention and postintervention pulse rates were not statistically different for the control and treatment groups (p = .0807). VAS postintervention mean for the spinal manipulation group was significantly lower than the control means (p = .05) and from its corresponding preintervention mean (p = .001). Spinal manipulation significantly decreased the intensity of emotional arousal reported by phobic subjects. The mechanism for this effect is not known.


Evaluation of the revised trauma and injury severity scores in elderly trauma patients.
Severity-of-illness scoring systems have primarily been developed for, and validated in, younger trauma patients. We sought to determine the accuracy of the injury severity score (ISS) and the revised trauma score (RTS) in predicting mortality and hospital length of stay (LOS) in trauma patients over the age of 65 treated in our emergency department (ED). Using the Illinois Trauma Registry, we identified all patients 65 years and older treated in our level I trauma facility from January 2004 to November 2007. The primary outcome was death; the secondary outcome was overall hospital length of stay (LOS). We measured associations between scores and outcomes with binary logistic and linear regression. A total of 347 patients, 65 years of age and older were treated in our hospital during the study period. Median age was 76 years (IQR 69-82), with median ISS 13 (IQR 8-17), and median RTS 7.8 (IQR 7.1-7.8). Overall mortality was 24%. A higher value for ISS showed a positive correlation with likelihood of death, which although statistically significant, was numerically small (OR=1.10, 95% CI 1.06 to 1.13, P<0.001). An elevated RTS had an inverse correlation to likelihood of death that was also statistically significant (OR=0.48, 95% CI 0.39 to 0.58, P<0.001). Total hospital LOS increased with increasing ISS, with statistical significance decreasing at the highest levels of ISS, but an increase in RTS not confirming the predicted decrease in total hospital LOS consistently across all ranges of RTS. The ISS and the RTS were better predictors of mortality than hypothesized, but had limited correlation with hospital LOS in elderly trauma patients. Although there may be some utility in these scores when applied to the elderly population, caution is warranted if attempting to predict the prognosis of patients.


Urinary tract infections. How to manage nursing home patients with or without chronic catheterization.
Urinary tract infections (UTIs)--including cystitis, pyelonephritis, and catheter-associated infections--are among the most common nursing home-acquired infections. Asymptomatic bacteriuria can be identified in 20 to 50% of nursing home residents who do not have bladder catheters and in 100% of those who do. Diagnostic tests for nursing home patients with suspected UTI include urinalysis, urine culture, and sensitivity testing. Treatment of cystitis can usually be managed in the nursing home with oral antibiotics. Initial therapy with a parenteral agent is often recommended in nursing home-acquired pyelonephritis.


Potential regulatory role of microRNAs in the development of bovine gastrointestinal tract during early life.
This study aimed to investigate the potential regulatory role of miRNAs in the development of gastrointestinal tract (GIT) during the early life of dairy calves. Rumen and small intestinal (mid-jejunum and ileum) tissue samples were collected from newborn (30 min after birth; n = 3), 7-day-old (n = 6), 21-day-old (n = 6), and 42-day-old (n = 6) dairy calves. The miRNA profiling was performed using Illumina RNA-sequencing and the temporal and regional differentially expressed miRNAs were further validated using qRT-PCR. Analysis of 16S rRNA gene copy numbers was used to quantify total bacteria, Bifidobacterium and Lactobacillus species. The expression of miR-143 was abundant in all three gut regions, at all time points and it targets genes involved primarily in the proliferation of connective tissue cells and muscle cells, suggesting a role in regulating rapid tissue development during the early life of calves. The expression of miR-146, miR-191, miR-33, miR-7, miR-99/100, miR-486, miR-145, miR-196 and miR-211 displayed significant temporal differences (FDR <0.05), while miR-192/215, miR-194, miR-196, miR-205 and miR-31 revealed significant regional differences (FDR <0.05). The expression levels of miR-15/16, miR-29 and miR-196 were positively correlated with the copy numbers of 16S rRNA gene of Bifidobacterium or Lactobacillus species or both (P<0.05). Functional analysis using Ingenuity Pathway Analysis identified the above mentioned differentially expressed miRNAs as potential regulators of gut tissue cell proliferation and differentiation. The bacterial density-associated miRNAs were identified as modulators of the development of lymphoid tissues (miR-196), maturation of dendritic cells (miR-29) and development of immune cells (miR-15/16). The present study revealed temporal and regional changes in miRNA expression and a correlation between miRNA expression and microbial population in the GIT during the early life, which provides further evidence for another mechanism by which host-microbial interactions play a role in regulating gut development.


Rasch analysis examining processing mechanisms of the object location memory test revised.
This study's objective was the construction and examination of the Object Location Memory Revised (OLM-R), an instrument designed to measure spatial memory. The OLM-R measured a participant's (N=111) ability to inspect a spatial array and recall image identities and positions after a distractor task. Rasch methodology and regression analyses were employed to explore the influence a priori design factors have on performance and item difficulty. Rasch analyses revealed that, while the OLM-R has a misfitting item, overall the instrument shows good measurement properties. Specific analyses examining complexity factors indicated that Object Manipulation (e.g., moving, replacing, or unchanging) and the Number of Items in an array were leading influences of OLM-R performance.


CSF multianalyte profile distinguishes Alzheimer and Parkinson diseases.
The therapeutic imperative for Alzheimer disease (AD) and Parkinson disease (PD) calls for discovery and validation of biomarkers. Increased cerebrospinal fluid (CSF) tau and decreased amyloid (A) beta42 have been validated as biomarkers of AD. In contrast, there is no validated CSF biomarker for PD. We validated our proteomics-discovered multianalyte profile (MAP) in CSF from 95 control subjects, 48 patients with probable AD, and 40 patients with probable PD. An optimal 8-member MAP agreed with expert diagnosis for 90 control subjects (95%), 36 patients with probable AD (75%), and 38 patients with probable PD (95%). This MAP consisted of the following (in decreasing order of contribution): tau, brain-derived neurotrophic factor, interleukin 8, Abeta42, beta2-microglobulin, vitamin D binding protein, apolipoprotein (apo) AII, and apoE. This first large-scale validation of a proteomic-discovered MAP suggests a panel of 8 CSF proteins that are highly effective at identifying PD and moderately effective at identifying AD.


Smarter Than Thou, Holier Than Thou: The Dynamic Interplay Between Cognitive and Moral Enhancement.
The debate about the desirability of using drugs to enhance human skills encompasses cognitive abilities such as memory and attention, and moral capacities such as emotional empathy and a sense of fairness. These two strands of literature in bioethics have grown relatively independent from each other, and an implicit framing assumption has emerged suggesting that apparently morally neutral cognitive capacities and paradigmatically moral capacities are distinct and vary independently of each other. Here, we identify key distinctions between competing accounts of cognitive enhancement and moral enhancement and argue that, despite the polarized nature of the bioethical debate, cognitive and moral capacities are intertwined. For example, moral behavior can be improved by enhancing "morally neutral" abilities such as attention span; and cognitive skills can be honed by means of socio-moral interaction. Further, cognitive skill is frequently assigned the abstract status of virtue and treated in the same way as more paradigmatically "moral" traits. We argue that the distinction between moral and cognitive enhancement is more apparent than real, since despite being nominally treated as distinct, cognitive and moral skills are frequently interdependent. As such we present evidence to support the claim that the enhancement of these two kinds of capacities cannot be clearly disaggregated from each other in the way that the theoretical poles of the debate in the literature suggest. We synthesize relevant scientific and bioethical literature and combine it with a line of analysis derived from Peter Hacker to show more clearly the terms of what can be said intelligibly about cognitive and moral skills and their enhancement. As a result of this analysis, we conclude that ethical questions in human bioenhancement are only fully intelligible at the level of persons imbued with feelings, thoughts, intentions, desires, values, and abilities, embedded within a particular social context, rather than at the level of pharmacological modulation of particular cognitive or affective capacities which, though conceptually distinguishable, in the embodied context of moral agency are profoundly intertwined.


Effects of bacterial vaginosis and other genital infections on the natural history of human papillomavirus infection in HIV-1-infected and high-risk HIV-1-uninfected women.
Whether the natural history of human papillomavirus (HPV) infection is affected by bacterial vaginosis (BV) or Trichomonas vaginalis (TV) infection has not been adequately investigated in prospective studies. Human immunodeficiency virus 1 (HIV-1)-infected (n=1763) and high-risk HIV-1-uninfected (n=493) women were assessed semiannually for BV (by Nugent's criteria), TV infection (by wet mount), type-specific HPV (by polymerase chain reaction with MY09/MY11/HMB01 HPV primers), and squamous intraepithelial lesions (SIL) (by cytological examination). Sexual history was obtained from patient report at each visit. Risk factors for prevalent and incident HPV infection and SIL were evaluated by use of multivariate models. BV was associated with both prevalent and incident HPV infection but not with duration of HPV infection or incidence of SIL. TV infection was associated with incident HPV infection and with decreased duration and lower prevalence of HPV infection. TV infection had no association with development of SIL. Effects of BV and TV infection were similar in HIV-1-infected and high-risk HIV-1-uninfected women. HIV-1 infection and low CD4(+) lymphocyte count were strongly associated with HPV infection and development of SIL. BV and TV infection may increase the risk of acquisition (or reactivation) of HPV infection, as is consistent with hypotheses that the local cervicovaginal milieu plays a role in susceptibility to HPV infection. The finding that BV did not affect persistence of HPV infection and that TV infection may shorten the duration of HPV infection helps explain the lack of effect that BV and TV infection have on development of SIL.


Physical activity and blood pressure in childhood: findings from a population-based study.
The pathological processes associated with development of cardiovascular disease begin early in life. For example, elevated blood pressure (BP) can be seen in childhood and tracks into adulthood. The relationship between physical activity (PA) and BP in adults is well-established, but findings in children have been inconsistent, with few studies measuring PA mechanically. Children aged 11 to 12 years were recruited from the Avon Longitudinal Study of Parents and Children. 5505 had systolic and diastolic BP measurements, plus valid (at least 10 hours for at least 3 days) accelerometer measures of PA; total PA recorded as average counts per minute (cpm) over the period of valid recording, and minutes per day spent in moderate to vigorous PA (MVPA). Data on a number of possible confounders were also available. Small inverse associations were observed; for systolic BP, beta=-0.44 (95% confidence interval -0.59, -0.28) mm Hg per 100 cpm, and beta=-0.66 (95% CI -0.92, -0.39) mm Hg per 15 minutes/d MVPA, adjusting for child's age and gender. After adjustment for potential confounders, associations were weakened but remained. When PA variables were modeled together, associations with total PA were only a little weaker, whereas those with MVPA were substantially reduced; for systolic BP, beta=-0.42 (95% CI -0.71, -0.13) mm Hg per 100 cpm, and beta=-0.03 (95% CI -0.54, 0.48) mm Hg per 15 minutes/d MVPA. In conclusion, higher levels of PA were associated with lower BP, and results suggested that the volume of activity may be more important than the intensity.


Injury and death in clinical trials and compensation: Rule 122 DAB.
Three amendments to the drugs and cosmetics rules were published in quick succession in 2013. These addressed the issues of compensation of injury and death in clinical trials in addition to the role and registration of Ethics Committees. Of the three, the first and the third make an impact on the clinical research activities in India. The second amendment has codified the conduct of clinical trials, putting together rules, which appeared in different sections of Schedule Y. The first amendment deals with the compensation for injuries and deaths taking place during clinical trials while the third deals with registration of Ethics Committees. Despite the long delay in the issue of compensation rules, there appears significant room for improvement. The most problematic are conditions of injury and death in which compensation has to be paid. When compared with other countries, the Indian rules seem unduly harsh on sponsors and are at significant variance with those in UK. The rules are very generous toward subjects and compensation is likely to become an alternative to insurance in terminally ill subjects. The implementation of these rules will make clinical trials in India more expensive and hurt the industry that is already struggling through other handicaps. There is an urgent need to make the the environment more industry friendly to attract more clinical work.


Age-Related Changes in Quantitative Strain of Mouse Astrocytic Lamina Cribrosa and Peripapillary Sclera Using Confocal Microscopy in an Explant Model.
The purpose of this study was to measure the full-field deformation response to IOP change in the peripapillary sclera (PPS) and astrocytic lamina cribrosa (ALC) of young and old mouse eyes ex vivo. Thirty-eight transgenic reporter mice with green fluorescent protein-expressing astrocytes were studied at 2 to 4 months and 13 to 15 months old. The ALC and PPS of the explant eyes were imaged using laser scanning microscopy under controlled inflation from 10 to 30 mm Hg. Strains were estimated for the ALC and PPS from imaged volumes using digital volume correlation. ALC strains were significantly greater than zero nasal-temporally for both age groups (mean = 4.3% and 4.0%; each P ≤ 0.004) and significantly greater than zero in the inferior-superior direction for younger mice (P = 0.0004). Younger mice had larger ALC inferior-superior strains than older mice (P = 0.002). The ALC area and perimeter enlarged with inflation in both age groups, with a greater increase in younger than in older mice (all P ≤ 0.004). The ALC nasal-temporal diameter change was greater than inferior-superiorly, and younger mice had greater enlargement nasal-temporally than older. PPS maximum shear strain was greater in the older mice (P = 0.002). The axial lengths of older mice were 14% longer and the PPS was 16% thinner than younger mice (both P = 0.0003). The behavior of the ALC in younger mice with inflation exhibited greater strains and enlargement of ALC area than older mice. Some strain measures in the PPS were greater in older mice, likely related to their longer axial length and thinner PPS.


Assessment of sorbent impregnated PUF disks (SIPs) for long-term sampling of legacy POPs.
Two field studies were conducted for one year using sorbent-impregnated polyurethane foam (SIP) disks for PCB and PBDE air sampling. SIP disks were introduced by Shoeib et al. (2008) as an alternative passive air sampling medium to the polyurethane foam (PUF) disk and have the advantage of a higher holding capacity for organic chemicals. The first study on SIP disks confirmed their application for measuring volatile perfluorinated compounds (PFCs) and their ability to maintain time-integrated (linear) air sampling. In this study, the suitability of the SIP disks for long-term sampling of polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and hexachlorobenzene (HCB) was assessed. SIP disks were deployed at a rural site in the UK and harvested after periods ranging from 35-350 days. Atmospheric POP concentrations were monitored with a high-volume air sampler during the deployment period. Linear uptake was observed for all monitored PCBs and PBDEs over the full exposure time. Air-sampler equilibrium was observed for HCB after 6 months. In a second field study, SIP disks were deployed for one year at 10 sites on a latitudinal transect in the UK and Norway, at which air sampling has been undertaken previously with different passive air sampling media since 1994. The estimated concentrations and spatial distributions derived from the SIP disks were largely in agreement with previously reported data.


2,5-Dimethyl-4-hydroxy-3(2H)-furanone as an Anti-biofilm Agent Against Non-Candida albicans Candida Species.
The predominance of non-Candida albicans Candida (NCAC) species causing healthcare-associated infections has increased over the last decade pertaining to their ability to form biofilms on medical devices. These biofilm-associated infections are challenging to treat as they are resistant to antifungal agents and evade host-immune response resulting in a high risk of device failure or biomaterial removal. Thus, to minimize the risk of biofilm-associated infections, preventing biofilm formation is the best approach which is mediated by the quorum quenching process. The present study investigated the modulatory effect of 2,5-dimethyl-4-hydroxy-3(2H)-furanone (DMHF) on NCAC biofilm formation and also assessed the effect of the DMHF-coated catheters on biofilm formation of NCAC. The NCAC isolates studied were Candida tropicalis, Candida glabrata and Candida krusei isolated from catheter tip, urine and blood, respectively. DMHF at a concentration of 30 µg/mL showed an inhibitory effect against NCAC biofilms at various stages and was statistically significant (p ≤ 0.05) against the various concentrations (50-5 µg/mL) tested and also among the three phases of experiment. The furanone content on coated catheters ranged from 170 to 750 µg and release of furanone from the coated catheter was about 15 µg for 30 days. The effect of DMHF-coated catheters on NCAC biofilm formation was observed by the scanning electron microscopy which revealed the absence of NCAC adherence on DMHF-coated catheters. This study provides a design to develop furanone-coated biomaterials which could be implemented in healthcare settings to reduce medical device-associated infections. The excellent biological performance, combined with their antimicrobial properties, suggests that 2,5-dimethyl-4-hydroxy-3(2H)-furanone could be an effective anti-infective coating for implantable devices.


Preoperative tumor size at MRI predicts deep myometrial invasion, lymph node metastases, and patient outcome in endometrial carcinomas.
The aim of this study was to explore the relation between preoperative tumor size based on magnetic resonance imaging (MRI) and the surgical pathologic staging parameters (deep myometrial invasion, cervical stroma invasion, and metastatic lymph nodes) and to assess the prognostic impact of tumor size in endometrial carcinomas. Interobserver variability for the different tumor size measurements was also assessed. Preoperative pelvic MRI of 212 patients with histologically confirmed endometrial carcinomas was read independently by 3 radiologists. Maximum tumor diameters were measured in 3 orthogonal planes (anteroposterior, transverse, and craniocaudal planes [CC]), and tumor volumes were estimated. Tumor size was analyzed in relation to surgical staging results and patient survival. The multivariate analyses were adjusted for preoperative risk status based on endometrial biopsy. Intraclass correlation coefficients and receiver operating characteristics curves for the different tumor measurements were also calculated. Anteroposterior tumor diameter independently predicted deep myometrial invasion (P < 0.001), whereas CC tumor diameter tended to independently predict lymph node metastases (P = 0.06). Based on receiver operating characteristic curves, the following tumor size cutoff values were identified: anteroposterior diameter greater than 2 cm predicted deep myometrial invasion (unadjusted odds ratio [OR], 12.4; P < 0.001; adjusted OR, 6.7; P < 0.001) and CC diameter greater than 4 cm predicted lymph node metastases (unadjusted OR, 6.2; P < 0.001; adjusted OR, 4.9; P = 0.009). Large tumor size was associated with reduced progression/recurrence-free survival (P ≤ 0.005 for all size parameters), and CC diameter had an independent impact on survival (adjusted hazards ratio, 1.04; P = 0.009). The interobserver variability for the different size measurements was very low (intraclass correlation coefficient, 0.78-0.85). Anteroposterior tumor diameter greater than 2 cm predicts deep myometrial invasion, and CC tumor diameter greater than 4 cm predicts lymph node metastases. Tumor size is a strong prognostic factor in endometrial carcinomas. Preoperative tumor measurements based on MRI may potentially improve preoperative risk stratification models and thus enable better tailored surgical treatment in endometrial cancer.


A genetic discontinuity in root-nodulating bacteria of cultivated pea in the Indian trans-Himalayas.
Evolutionary relationships of 120 root-nodulating bacteria isolated from the nodules of Pisum sativum cultivated at 22 different locations of the trans-Himalayan valleys of Lahaul and Spiti in the state of Himachal Pradesh of India were studied using 16S rRNA gene PCR-RFLP, ERIC-PCR, sequencing of 16S rRNA, atpD, recA, nodC and nifH genes, carbon-source utilization pattern (BIOLOG™), and whole-cell fatty acid profiling. The results demonstrated that all isolates belonged to Rhizobium leguminosarum symbiovar viciae (Rlv). Isolates from the two valleys were clearly separated on the basis of ERIC fingerprints, carbon-source utilization pattern, and whole-cell fatty acid methyl esters. Phylogenetic analysis of atpD, recA, nodC and nifH genes revealed a common Rlv sublineage in Spiti valley. Lahaul valley isolates were represented by three sequence types of atpD and recA genes, and four sequence types of nodC and nifH genes. Genotypes from the two valleys were completely distinct, except for two Lahaul isolates that shared nodC and nifH sequences with Spiti isolates but were otherwise more similar to other Lahaul isolates. Isolates from the two highest Spiti valley sites (above 4000 m) had a distinctive whole-cell fatty acid profile. Spiti valley isolates are closely related to Rlv sublineages from Xinjiang and Shanxi provinces in China, while Lahaul valley isolates resemble cosmopolitan strains of the western world. The high mountain pass between these valleys represents a boundary between two distinct microbial populations.


Insecticide resistance in malaria vectors along the Thailand-Myanmar border.
There is a paucity of data about the susceptibility status of malaria vectors to Public Health insecticides along the Thailand-Myanmar border. This lack of data is a limitation to guide malaria vector-control in this region. The aim of this study was to assess the susceptibility status of malaria vectors to deltamethrin, permethrin and DDT and to validate a simple molecular assay for the detection of knock-down resistance (kdr) mutations in the study area. Anopheles mosquitoes were collected in four sentinel villages during August and November 2014 and July 2015 using human landing catch and cow bait collection methods. WHO susceptibility tests were carried out to measure the mortality and knock-down rates of female mosquitoes to deltamethrin (0.05%), permethrin (0.75%) and DDT (4%). DNA sequencing of a fragment of the voltage-gated sodium channel gene was carried out to identify knock-down resistance (kdr) mutations at position 1014 in mosquitoes surviving exposure to insecticides. A total of 6295 Anopheles belonging to ten different species were bioassayed. Resistance or suspected resistance to pyrethroids was detected in An. barbirostris (s.l.) (72 and 84% mortality to deltamethrin (n = 504) and permethrin (n = 493) respectively), An. hyrcanus (s.l.) (33 and 48% mortality to deltamethrin (n = 172) and permethrin (n = 154), respectively), An. jamesii (87% mortality to deltamethrin, n = 111), An. maculatus (s.l.) (85 and 97% mortality to deltamethrin (n = 280) and permethrin (n = 264), respectively), An. minimus (s.l.) (92% mortality, n = 370) and An. vagus (75 and 95% mortality to deltamethrin (n =148) and permethrin (n = 178), respectively). Resistance or suspected resistance to DDT was detected in An. barbirostris (s.l.) (74% mortality, n = 435), An. hyrcanus (s.l.) (57% mortality, n = 91) and An. vagus (97% mortality, n = 133). The L1014S kdr mutation at both heterozygous and homozygous state was detected only in An. peditaeniatus (Hyrcanus Group). Resistance to pyrethroids is present along the Thailand-Myanmar border, and it represents a threat for malaria vector control. Further investigations are needed to better understand the molecular basis of insecticide resistance in malaria vectors in this area.


forked proteins are components of fiber bundles present in developing bristles of Drosophila melanogaster.
The forked (f) gene of Drosophila melanogaster encodes six different transcripts 6.4, 5.6, 5.4, 2.5, 1.9, and 1.1 kb long. These transcripts arise by the use of alternative promoters. A polyclonal antibody raised against a domain common to all of the forked-encoded products has been used to identify forked proteins on two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and in Drosophila pupal tissues. The antibody stains fiber bundles present in bristle cells for about 15 hr during normal pupal development. Electron microscopy shows that these fibers are present from 40 to 53 hr in bristles of wild-type flies but are absent in the null f36a mutant. The forked protein(s) thus appear to be an essential part of the bristle fibers. The phenotype of the f36a mutation can be rescued by a 13-kb fragment of the forked locus containing the coding regions for the 2.5, 1.9, and 1.1-kb transcripts, suggesting that the proteins encoded by the three large forked RNAs are dispensable during bristle development. Increasing the copy number of a P[w+,f+] construct containing the 13-kb fragment induces a hypermorphic bristle phenotype whose severity correlates with the number of copies of P[w+,f+] present. These results indicate that alterations in the ratios among the forked proteins, or between forked products and other components of the fiber, result in abnormal assembly of the fibrillar cytoplasmic structures necessary for bristle morphogenesis.


Factors in glucocorticoid regimens associated with treatment response and relapses of IgG4-related disease: a multicentre study.
Glucocorticoids (GC) are effective for treating IgG4-related disease (IgG4-RD); however, relapse is often observed. We conducted a retrospective multicentre study to investigate risk factors in GC regimens associated with relapses of IgG4-RD. Data on 166 patients with definitive IgG4-RD diagnosis were collected from 12 institutions. Comprehensive surveillance of clinical backgrounds and GC regimens as well as multivariate analysis of factors associated with treatment responses and relapses was performed. To determine the initial maximal GC dose, the patients were stratified into three groups depending on the initial prednisolone (PSL) dosage: <0.39, 0.4-0.69 and >0.7 mg/kg/day. The multivariate analysis extracted the disease duration and reduction speed of initial GC dose. Patients treated with initial GC <0.39 or >0.7 mg/kg/day of PSL showed higher relapse rates than those treated with 0.4-0.69 mg/kg/day. The relapse rates were significantly higher in patients with fast reduction of the initial dose (>0.4 mg/day) than in patients with slow reduction (<0.4 mg/day). To avoid relapse, 0.4-0.69 mg/kg/day of initial PSL with slow reduction speed (<0.4 mg/day) is needed in the early treatment of IgG4-RD.


Ages, irradiation history, and chemical composition of lunar rocks from the sea of tranquillity.
The (87)Rb-(87)Sr internal isochrons for five rocks yield an age of 3.65 +/-0.05 x 10(9) years which presumably dates the formation of the Sea of Tranquillity. Potassium-argon ages are consistent with this result. The soil has a model age of 4.5 x10(9) years, which is best regarded as the time of initial differentiation of the lunar crust. A peculiar rock fragment from the soil gave a model age of 4.44 x 10(9) years. Relative abundances of alkalis do not suggest differential volatilization. The irradiation history of lunar rocks is inferred from isotopic measurements of gadolinium, vanadium, and cosmogenic rare gases. Spallation xenon spectra exhibit a high and variable (131)Xe/(126)Xe ratio. No evidence for (129)I was found. The isotopic composition of solar-wind xenon is distinct from that of the atmosphere and of the average for carbonaceous chondrites, but the krypton composition appears similar to average carbonaceous chondrite krypton.


A chromosome-based draft sequence of the hexaploid bread wheat (Triticum aestivum) genome.
An ordered draft sequence of the 17-gigabase hexaploid bread wheat (Triticum aestivum) genome has been produced by sequencing isolated chromosome arms. We have annotated 124,201 gene loci distributed nearly evenly across the homeologous chromosomes and subgenomes. Comparative gene analysis of wheat subgenomes and extant diploid and tetraploid wheat relatives showed that high sequence similarity and structural conservation are retained, with limited gene loss, after polyploidization. However, across the genomes there was evidence of dynamic gene gain, loss, and duplication since the divergence of the wheat lineages. A high degree of transcriptional autonomy and no global dominance was found for the subgenomes. These insights into the genome biology of a polyploid crop provide a springboard for faster gene isolation, rapid genetic marker development, and precise breeding to meet the needs of increasing food demand worldwide.


Insights into molecular features of Venerupis decussata oocytes: a microarray-based study.
The production of Venerupis decussata relies on wild seed collection, which has been recently compromised due to recruitment failure and severe mortalities. To address this issue and provide an alternative source of seed, artificial spawning and larval rearing programs were developed. However, hatchery-based seed production is a relatively new industry and it is still underdeveloped. A major hurdle in the European clam seed production is the control of spawning and reproduction, which is further hindered by the impossibility of obtaining fertile gametes by gonadal "stripping", as meiosis re-initiation is constrained to a maturation process along the genital ducts. In the present study, oocytes were collected from 15 females and microarray analyses was performed to investigate gene expression profiles characterizing released and stripped ovarian oocytes. A total of 198 differentially expressed transcripts between stripped and spawned oocytes were detected. Functional analysis carried out on these transcripts highlighted the importance of a few biological processes, which are most probably implicated in the control of oocyte competence. Significant differences were observed for transcripts encoding proteins involved in meiosis progression (e.g. dual specificity phosphatase CDC25), WNT signalling (e.g. frizzled class receptor 8, wingless-type MMTV integration site family member 4), steroid synthesis (e.g. progestin and adipoQ receptor family member 3, cytochrome P450-C17), mRNA processing (e.g. zinc finger protein XlCOF28), calcium regulation (e.g. regucalcin, calmodulin) and ceramide metabolism (ceramidase B, sphingomyelinase). This study provides new information on transcriptional profiles putatively associated with ovarian egg infertility, and suggests potential mechanisms regulating early oocyte development in clams. Genes which were differentially expressed between stripped and spawned oocytes might have a pivotal role during maturation process in the gonadal duct and could be interesting targets for further functional studies aiming to make ovarian oocytes fertilizable.


The "most wanted" taxa from the human microbiome for whole genome sequencing.
The goal of the Human Microbiome Project (HMP) is to generate a comprehensive catalog of human-associated microorganisms including reference genomes representing the most common species. Toward this goal, the HMP has characterized the microbial communities at 18 body habitats in a cohort of over 200 healthy volunteers using 16S rRNA gene (16S) sequencing and has generated nearly 1,000 reference genomes from human-associated microorganisms. To determine how well current reference genome collections capture the diversity observed among the healthy microbiome and to guide isolation and future sequencing of microbiome members, we compared the HMP's 16S data sets to several reference 16S collections to create a 'most wanted' list of taxa for sequencing. Our analysis revealed that the diversity of commonly occurring taxa within the HMP cohort microbiome is relatively modest, few novel taxa are represented by these OTUs and many common taxa among HMP volunteers recur across different populations of healthy humans. Taken together, these results suggest that it should be possible to perform whole-genome sequencing on a large fraction of the human microbiome, including the 'most wanted', and that these sequences should serve to support microbiome studies across multiple cohorts. Also, in stark contrast to other taxa, the 'most wanted' organisms are poorly represented among culture collections suggesting that novel culture- and single-cell-based methods will be required to isolate these organisms for sequencing.