Sesame seeds essential oil and Sesamol modulate the pro-inflammatory function of macrophages and dendritic cells and promote Th2 response.
Background: Herbal medicine is becoming progressively accepted treatment for management of different diseases worldwide. Recognition of the active ingredients and mechanisms of herbal medicine against the immune system and related anomalies is highly favorable. This experimental study aimed to investigate the effects of Sesame (Sesamum indicum L.) essential oil and sesamol as effective components on mouse splenocytes subsets, macrophages and dendritic cells (DCs). Methods: Effective components of sesame were extracted and used to treat splenocytes, PHA (5μg/ml) and LPS (10 μg/ml) stimulated splenocytes, macrophages and DCs in different concentration (0.01-100 μg/ml). The cell proliferation/viability was measured using the MTT assay and nitrite levels were measured by the diazotization method. Moreover, TNF-α and IL-1β cytokines concentration were assayed by ELISA. Treated DCs also analysed for maturation marker levels and cytokine production. Results: Analysis of the results indicated that sesame components suppress PHA-stimulated splenocytes with no effect on LPS-stimulated subsets. Furthermore, the sesame ingredients reduced the release of IFN-γ and increased secretion of IL-4 from lymphocytes. Macrophages viability was not affected and production of NO, TNF-α, and IL-1β were inhibited using sesame essential oil and sesamol. DCs phenotype skewed to immature and release of TNF-α and IL-1β were abrogated form DCs. Conclusion: These results indicate that sesame essential oil and its effective component as sesamol may capable of suppressing the response of cellular immunity with the domination of Th2 responses and also could modulate macrophages and the dendritic cells proinflammatory functions.


An early Spanish vocabulary for children who use AAC: developmental and linguistic considerations.
For children with complex communication needs in the early stages of language development, access to appropriate vocabulary provides a means for social interaction and participation, and the foundation for the acquisition of grammar and other language related skills. While numerous resources are available to support decision making for speakers of English, there is a pressing need to rapidly expand such resources for other languages. Spanish is the official language in 20 countries, and in other countries (e.g., United States) Spanish-speaking communities represent a substantial proportion of the population. The aim of this study was to produce a developmentally-relevant word list for use by Spanish-speaking children in the early stages of language development. The list was developed from an analysis of overlap between published and validated lists of words produced by young Spanish speaking children with typical development. The list includes a wide range of word classes and semantic categories and is proposed as a tool to assist professionals, families and software developers in the process of selecting an initial lexicon for children who require AAC and are learning Spanish. Implications of our findings for vocabulary selection and future research directions are discussed.


Synthesis of nanostructured metal-organic films: surface X-ray radiolysis of silver ions using a langmuir monolayer as a template.
An application of the radiolysis method using an X-ray synchrotron beam is developed as a novel approach to the synthesis of metal-organic films with controlled shapes and thickness. We demonstrate that a Langmuir monolayer deposited onto a silver ion containing subphase, irradiated by an incident beam impinging below the critical angle for total reflection, induces the synthesis of a stable nanostructured silver-organic ultrathin film at the air-water interface. The X-ray scattering is also used to monitor in situ the structure of the silver layer during the synthesis process. The layer is observed by atomic force microscopy after its transfer onto a silicon substrate. One observes a film thickness of 4.6 nm, in good agreement with the X-ray penetration depth, about 4.5 nm. The silver structure is oriented by the initial organic film phase. This experiment demonstrates the considerable potential of this approach to produce various controlled metal-organic films with a surfactant self-assembly as a template.


Frequency of eating and concentrations of serum cholesterol in the Norfolk population of the European prospective investigation into cancer (EPIC-Norfolk): cross sectional study.
To examine the relation between self reported eating frequency and serum lipid concentrations in a free living population. Cross sectional population based study. Norfolk, England. 14 666 men and women aged 45-75 years from the Norfolk cohort of the European prospective investigation into cancer (EPIC-Norfolk). Concentrations of blood lipids. Mean concentrations of total cholesterol and low density lipoprotein cholesterol decreased in a continuous relation with increasing daily frequency of eating in men and women. No consistent relation was observed for high density lipoprotein cholesterol, body mass index, waist to hip ratio, or blood pressure. Mean cholesterol concentrations differed by about 0.25 mmol/l between people eating more than six times a day and those eating once or twice daily; this difference was reduced to 0.15 mmol/l after adjustment for possible confounding variables, including age, obesity, cigarette smoking, physical activity, and intake of energy and nutrients (alcohol, fat, fatty acids, protein, and carbohydrate). Concentrations of total cholesterol and low density lipoprotein cholesterol are negatively and consistently associated with frequency of eating in a general population. The effects of eating frequency on lipid concentrations induced in short term trials in animals and human volunteers under controlled laboratory conditions can be observed in a free living general population. We need to consider not just what we eat but how often we eat.


Nanodiscs for immobilization of lipid bilayers and membrane receptors: kinetic analysis of cholera toxin binding to a glycolipid receptor.
Nanodiscs are self-assembled soluble discoidal phospholipids bilayers encirculated by an amphipathic protein that together provide a functional stabilized membrane disk for the incorporation of membrane-bound and membrane-associated molecules. The scope of the present work is to investigate how nanodiscs and their incorporated membrane receptors can be attached to surface plasmon resonance sensorchips and used to measure the kinetics of the interaction between soluble molecules and membrane receptors inserted in the bilayer of nanodiscs. Cholera toxin and its glycolipid receptor G(M1) constitute a system that can be considered a paradigm for interactions of soluble proteins with membrane receptors. In this work, we have investigated different technologies for capturing nanodiscs containing the glycolipid receptor G(M1) in lipid bilayers, enabling measurements of binding of its soluble interaction partner cholera toxin B subunit to the receptor with the sensorchip-based surface plasmon resonance (SPR) technology. The measured stoichiometric and kinetic values of the interaction are in agreement with those reported by previous studies, thus providing proof-of-principle that nanodiscs can be employed for kinetic SPR studies.


Positional effect of preoperative neuronavigational magnetic resonance image on accuracy of posterior fossa lesion localization.
The aim of this study was to analyze the positional effect of MRI on the accuracy of neuronavigational localization for posterior fossa (PF) lesions when the operation is performed with the patient in the prone position. Ten patients with PF tumors requiring surgery in the prone position were prospectively enrolled in the study. All patients underwent preoperative navigational MRI in both the supine and prone positions in a single session. Using simultaneous intraoperative registration of the supine and prone navigational MR images, the authors investigated the images' accuracy, spatial deformity, and source of errors for PF lesions. Accuracy was determined in terms of differences in the ability of the supine and prone MR images to localize 64 test points in the PF by using a neuronavigation system. Spatial deformities were analyzed and visualized by in-house-developed software with a 3D reconstruction function and spatial calculation of the MRI data. To identify the source of differences, the authors investigated the accuracy of fiducial point localization in the supine and prone MR images after taking the surface anatomy and age factors into consideration. Neuronavigational localization performed using prone MRI was more accurate for PF lesions than routine supine MRI prior to prone position surgery. Prone MRI more accurately localized 93.8% of the tested PF areas than supine MRI. The spatial deformities in the neuronavigation system calculated using the supine MRI tended to move in the posterior-superior direction from the actual anatomical landmarks. The average distance of the spatial differences between the prone and supine MR images was 6.3 mm. The spatial difference had a tendency to increase close to the midline. An older age (> 60 years) and fiducial markers adjacent to the cervical muscles were considered to contribute significantly to the source of differences in the positional effect of neuronavigation (p < 0.001 and p = 0.01, respectively). This study demonstrated the superior accuracy of neuronavigational localization with prone-position MRI during prone-position surgery for PF lesions. The authors recommend that the scan position of the neuronavigational MRI be matched with the surgical position for more precise localization.


Pregnant teenagers' group: contributions to prenatal care.
To describe changes in nurses' care following the implementation of a group of pregnant teenagers in prenatal care based on the expectations and experiences of pregnant teenagers. Qualitative and descriptive study conducted from February to November 2013 at a Primary Care Unit in Fortaleza, Ceará, Brazil, through focus groups with 16 adolescents from the group of pregnant women in the second or third trimester of pregnancy. The analysis identified central ideas and units of meanings that formed the categories. The strategy of a group of pregnant teenagers, which provides a space for coexistence and the establishment of ties encourages these individuals to talk about their needs, re-signifying their ties. Educational strategies to promote self-care of pregnant teenagers and care for their babies involve the sharing of experiences, doubts and beliefs. Considerations and suggestions of the adolescents contributed to guide nurses' practice and provide a strategic space of care and support for pregnant adolescents in primary care.


Quantum effect on thermally activated glide of dislocations.
Crystal plasticity involves the motion of dislocations under stress. So far, atomistic simulations of this process have predicted Peierls stresses, the stress needed to overcome the crystal resistance in the absence of thermal fluctuations, of more than twice the experimental values, a discrepancy best-known in body-centred cubic crystals. Here we show that a large contribution arises from the crystal zero-point vibrations, which ease dislocation motion below typically half the Debye temperature. Using Wigner's quantum transition state theory in atomistic models of crystals, we found a large decrease of the kink-pair formation enthalpy due to the quantization of the crystal vibrational modes. Consequently, the flow stress predicted by Orowan's law is strongly reduced when compared with its classical approximation and in much closer agreement with experiments. This work advocates that quantum mechanics should be accounted for in simulations of materials and not only at very low temperatures or in light-atom systems.


From stem cell to red cell: regulation of erythropoiesis at multiple levels by multiple proteins, RNAs, and chromatin modifications.
This article reviews the regulation of production of RBCs at several levels. We focus on the regulated expansion of burst-forming unit-erythroid erythroid progenitors by glucocorticoids and other factors that occur during chronic anemia, inflammation, and other conditions of stress. We also highlight the rapid production of RBCs by the coordinated regulation of terminal proliferation and differentiation of committed erythroid colony-forming unit-erythroid progenitors by external signals, such as erythropoietin and adhesion to a fibronectin matrix. We discuss the complex intracellular networks of coordinated gene regulation by transcription factors, chromatin modifiers, and miRNAs that regulate the different stages of erythropoiesis.


Effect of occupational co-exposure to lead and cadmium on selected immunomodulatory cytokines.
Occupational exposure to heavy metals like lead (Pb) and cadmium (Cd) is associated with the development of several diseases. The objective of this study was to determine the effect of occupational co-exposure to Pb and Cd on the blood levels of selected immune-modulatory cytokines related to T helper (Th), that is, Th1, interleukin-2 (IL-2), Th2, (IL-4 and IL-10), and Th17, (IL-17) cells. The study comprised 207 individuals divided into two groups: exposed (n = 110) and nonexposed (n = 97). Blood Pb and Cd were determined using Graphite Furnace Atomic Absorption Spectroscopy, and serum levels of cytokines were measured using enzyme-linked immunosorbent assay (ELISA). The study revealed significantly higher blood Pb and Cd levels in the exposed group. A significant decrease in Th1 cytokine-IL-2 and Th2 cytokine-IL-10 was found, while IL-4 (Th2 cytokine) and IL-17 (Th17) levels were higher in the exposed group. In the mixed exposure analysis, among all the selected cytokines, IL-4 levels were significantly different between individuals having higher levels of both Pb and Cd versus lower levels of Pb and Cd. While IL-2 levels were highest among the low Pb and Cd group, the IL-17 levels were highest among individuals with higher Cd levels. The study demonstrated that co-exposure to low levels of Pb and Cd might have an immune-modulatory effect. The data suggested a metal-induced pro-inflammatory immune response.


[1-13C; methyl-2H3]methionine kinetics in humans: methionine conservation and cystine sparing.
Methionine (Met) conservation in healthy young adult men (4/diet group) was explored by supplying one of the following three L-amino acid based diets: 1) adequate Met but no cystine; 2) neither Met nor cystine; or 3) no Met but cystine supplementation. After 5 days, subjects received a continuous intravenous infusion of L-[1-13C; methyl-2H3]Met for 5 h while the diet was given as small isocaloric isonitrogenous meals. Estimates were made of rates of Met incorporation into protein synthesis (S) and release from body proteins (B), transmethylation (TM), remethylation of homocysteine (RM), and transsulfuration (TS). For the adequate Met diet, the rates were S = 24 +/- 2, B = 18 +/- 1, TM = 12.4 +/- 1.7, RM = 4.7 +/- 1.1, and TS = 7.6 +/- 0.6 (SE) mumol.kg-1.h-1. The sulfur amino acid-devoid diet significantly (P less than 0.05) reduced S, TM, RM, and TS. Supplementation of this diet with cystine reduced Met oxidation (P = 0.05). Therefore, two loci are quantitatively important regulatory points in Met conservation in vivo: 1) the distribution of Met between the pathways of protein anabolism and TM (Met locus) and 2) the distribution of homocysteine between RM and TS (homocysteine locus).


Anthropometric approximation of body weight in unresponsive stroke patients.
Thrombolysis of acute ischaemic stroke is based strictly on body weight to ensure efficacy and to prevent bleeding complications. Many candidate stroke patients are unable to communicate their body weight, and there is often neither the means nor the time to weigh the patient. Instead, weight is estimated visually by the attending physician, but this is known to be inaccurate. Based on a large general population sample of nearly 7000 subjects, we constructed approximation formulae for estimating body weight from simple anthropometric measurements (body height, and waist and hip circumference). These formulae were validated in a sample of 178 consecutive inpatients admitted to our stroke unit, and their accuracy was compared with the best visual estimation of two experienced physicians. The simplest formula gave the most accurate approximation (mean absolute difference 3.1 (2.6) kg), which was considerably better than the best visual estimation (physician 1: 6.5 (5.2) kg; physician 2: 7.4 (5.7) kg). It reduced the proportion of weight approximations mismatched by >10% from 31.5% and 40.4% (physicians 1 and 2, respectively) to 6.2% (anthropometric approximation). Only the patient's own estimation was more accurate (mean absolute difference 2.7 (2.4) kg). By using an approximation formula based on simple anthropometric measurements (body height, and waist and hip circumference), it is possible to obtain a quick and accurate approximation of body weight. In situations where the exact weight of unresponsive patients cannot be ascertained quickly, we recommend using this approximation method rather than visual estimation.


Ethanol consumption modifies the body turnover of cadmium: a study in a rat model of human exposure.
Ethanol (Et) abusers may also be exposed to excessive amounts of cadmium (Cd). Thus, the study was aimed at estimating the influence of Et on the body turnover of Cd in a rat model reflecting excessive alcohol consumption in humans chronically exposed to moderate and relatively high levels of this metal. For this purpose, Cd apparent absorption, retention in the body and concentration in the blood, stomach, duodenum, liver, kidney, spleen, brain, heart, testis and femur as well as its fecal and urinary excretion in the rats exposed to 5 and 50mg Cd l(-1) (in drinking water; for 16 weeks from the fifth week of the animal's life) and/or Et (5 g kg(-1) b.w. per 24 h, by oral gavage; for 12 weeks from the ninth week of life) were estimated. Moreover, the duodenal, liver and kidney pool of the nonmetallothionein (Mt)-bound Cd was evaluated. The administration of Et during the exposure to 5 or 50mg Cd l(-1) increased Cd accumulation in the gastrointestinal tract and its urinary excretion, and decreased Cd concentration in the blood, femur and numerous soft tissues (including liver and kidney) as well as the total pool of this metal in internal organs. Et modified or not the pool of the non-Mt-bound Cd, depending on the level of treatment with this metal. The results show that excessive Et consumption during Cd exposure may decrease the body burden of this metal, at least partly, by its lower absorption and increased urinary excretion. Based on this study, it can be concluded that Cd concentration in the blood and tissues of alcohol abusers chronically exposed to moderate and relatively high levels of this metal may be lower, whereas its urinary excretion is higher than in their nondrinking counterparts. However, since Et is toxic itself, the decreased body burden of Cd owing to alcohol consumption does not allow for the conclusion that the risk of health damage may be lower at co-exposure to these xenobiotics. In a further study, it will be investigated how the Et-induced changes in the body status of Cd influence the effects of its toxic action.


Ethylene as a Possible Mediator of Light- and Nitrate-Induced Inhibition of Nodulation of Pisum sativum L. cv Sparkle.
Exposure of pea (Pisum sativum L. cv Sparkle) roots to light suppressed nodulation and induced an increase in ethylene production by roots. Dim light did not affect the number of infections per centimeter of root on the primary root, but most infections were blocked when the infection thread was in the epidermis or in the outer cortex. This is the same stage of infection on lateral roots that is blocked by exogenous ethylene. Silver, an inhibitor of ethylene action, increased nodule number on roots exposed to dim light. Exposure of pea roots to nitrate also suppressed nodulation and induced increased ethylene production by roots. However, induction of ethylene production from roots by nitrate was less than that induced by dim light. Nitrate decreased the number of infections per centimeter of lateral root. With 25 mm nitrate, about half of the infections that occurred were blocked in the epidermis or in the outer cortex. Silver did not reverse the inhibitory effects of nitrate on nodulation. Our data indicate that the inhibitory effect of light may occur via increased ethylene production, but they do not support the hypothesis that ethylene mediates nitrate control of nodule number. The possible role of ethylene in regulating nodule number should be studied in experiments in which light is excluded from the roots.


MR Defecating Proctography with Emphasis on Posterior Compartment Disorders.
MR defecating proctography (MRDP) is a noninvasive examination that can be used for evaluating posterior compartment disorders. MRDP has several advantages over conventional fluoroscopic defecography. These benefits include high-contrast resolution evaluation of the deep pelvic organs, simultaneous multicompartmental assessment that is performed statically and dynamically during defecation, and lack of ionizing radiation. MRDP also provides a highly detailed anatomic evaluation of the pelvic floor supportive structures, including direct assessment of the pelvic floor musculature and indirect assessment of the endopelvic fascia. As the breadth of knowledge regarding anatomic and functional posterior compartment disorders expands, so too does the advancement of noninvasive and surgical treatment options for these conditions. High-quality MRDP examinations, with key anatomic and functional features reported, guide treatment planning. Reporting of MRDP examination findings with use of standardized terminology that emphasizes objective measurements rather than subjective grading aids consistent communication among radiologists, clinicians, and surgeons. Familiarity with commonly encountered posterior compartment pelvic floor pathologic entities that contribute to posterior compartment disorders and awareness of the essential information needed by surgeons are key to providing an optimal multidisciplinary discussion for planning pelvic floor dysfunction treatment. The authors provide an overview of the basic concepts of the MRDP acquisition technique, the anatomic abnormalities of posterior compartment pelvic floor pathologic entities associated with defecatory disorders, and recently developed interdisciplinary MRDP reporting templates and lexicons. In addition, the associated imaging findings that are key for surgical treatment guidance are highlighted. © RSNA, 2022 Online supplemental material is available for this article.


Multiscale simulation of the ground and photo-induced charge-separated states of a molecular triad in polar organic solvent: exploring the conformations, fluctuations, and free energy landscapes.
The approach of a multiscale simulation that combines quantum chemical calculations, classical molecular dynamics simulations, and statistical physics to integrate the information of the electronic states of a conformationally complex molecule into its structural distribution over an ensemble was performed to understand the influence of a polar organic solvent on the structural stability of carotene-porphyrin-fullerene molecular triad in both the ground and the photoinduced charge-separated excited states. The excited states of the triad were computed with the ab initio quantum chemical calculations using the algebraic diagrammatic construction through the second order correction (ADC(2)) method and the time-dependent density functional theory (TDDFT). The replica exchange molecular dynamics was used for the enhanced sampling of the ensemble in order to explore the phase space of the ground state and the photoinduced charge-separated excited state of triad in explicit tetrahydrofuran (THF) solvent. An importance sampling was strategically employed to bridge the gap between the two computational methods that aim to explore distinct realms of dynamics. We analyzed the free energy landscape, the structural fluctuations, the solvent arrangements, the static dielectric constant, and the interactions between the triad and the solvent molecules. The analysis of the free energy landscape of the triad indicates that the charge-separated excited state of the triad is thermodynamically stable in a linearly extended geometry, while the ground-state triad explores several extended and bent conformations that are populated in the local free energy minima separated by low free energy barriers at an order of thermal fluctuation (k(B)T). The stabilization of a linearly extended structure of the charge-separated excited state triad is predominantly due to the solvation interactions (van der Waals and electrostatic interactions) between the triad and the THF molecules as well as the interactions within the THF molecules. Differences in the charge distribution on a molecular triad induce slight changes in the dielectric property of THF near the triad. Our study suggests that by measuring the differences in a dielectric response of solvent near the triad, it is possible to provide insight into the population of the charge-separated electronic state of the triad relative to that of the ground state.


Vocational rehabilitation, interagency collaboration and social representations.
The aim of this study is to describe and analyse two important dimensions of vocational rehabilitation for disadvantaged groups and persons with disabilities: interagency collaboration and social representations. Four focus group discussions were conducted. The participants were 20 officials of various agencies who had taken part in collaboration projects in vocational rehabilitation. Qualitative content analysis was used for the analysis. The material was categorised and central themes identified. Three themes emerged: 'Collaboration Process', 'Other Agencies' and 'Object for Collaboration'. The results indicate that interagency collaboration is very important in vocational rehabilitation, but that there are a number of obstacles to smooth collaboration. The professionals of the different agencies shared social representations to a great extent. Working with people with psychiatric disorders is especially challenging, and conflicts tended to arise between the projects and the home organisations. Recognition of others' knowledge and respectfulness toward other professions facilitated vocational rehabilitation and the interagency collaboration process. The agencies' lack of flexibility increased the risk of conflicts as attempts were made to integrate the new working methods developed within the projects into the ordinary activities of the agencies.


Developing appropriate community-based postgraduate training in a developing country.
The School of Medical Sciences, KNUST, established in 1975, indicated its educational orientation to a community-based and community-oriented system using the problem-based learning approach. Falling victim to the phenomenon of brain-drain, the School developed a postgraduate medical curriculum which takes into account the needs and demands of national governments and communities for quality specialist care and equitable distribution of existing health facilities. There is an innovative community-based fourth year for obstetrics and gynaecology, ophthalmology, otorhinolaryngology, paediatrics, surgery and medicine. After attainment of the Part I Examination, residents are sent to Ulm, Maastricht, Britain and the United States for clinical attachments for a period of 3 to 6 months before returning to sit for their Final Part II for the Fellowship of the West African College of Surgeons or Physicians (FWACS, FWACP). By ensuring that the Final Examinations take place after the elective attachment, none of the new breed of specialists so far produced has been lost to the Western world. The strengths and weaknesses of the programme are discussed.


Does random treatment assignment cause harm to research participants?
Some argue that by precluding individualized treatment, randomized clinical trials (RCTs) provide substandard medical care, while others claim that participation in clinical research is associated with improved patient outcomes. However, there are few data to assess the impact of random treatment assignment on RCT participants. We therefore performed a systematic review to quantify the differences in health outcomes between randomized trial participants and eligible non-participants. Studies were identified by searching Medline, the Web of Science citation database, and manuscript references. Studies were eligible if they documented baseline characteristics and clinical outcomes of RCT participants and eligible non-participants, and allowed non-participants access to the same interventions available to trial participants. Primary study outcomes according to patient group (randomized trial participants versus eligible non-participants) were extracted from all eligible manuscripts. For 22 of the 25 studies (88%) meeting eligibility criteria, there were no significant differences in clinical outcomes between patients who received random assignment of treatment (RCT participants) and those who received individualized treatment assignment (eligible non-participants). In addition, there was no relation between random treatment assignment and clinical outcome in 15 of the 17 studies (88%) in which randomized and nonrandomized patients had similar health status at baseline. These findings suggest that randomized treatment assignment as part of a clinical trial does not harm research participants.


In vitro and in vivo evidence of a potential A(H1N1)pdm09 antigenic drift mediated by escape mutations in the haemagglutinin Sa antigenic site.
Influenza A(H1N1)pdm09 virus continues to circulate worldwide without evidence of significant antigenic drift between 2009 and 2016. By using escape mutants, we previously identified six haemagglutinin (HA) changes (T80R, G143E, G158E, N159D, K166E and A198E) that were located within antigenic sites. Combinations of these mutations were introduced into the A(H1N1)pdm09 HA plasmid by mutagenesis. Reassortant 6 : 2 viruses containing both the HA and NA genes of the A(H1N1)pdm09 and the six internal gene segments of A/PR/8/34 were rescued by reverse genetics. In vitro, HA inhibition and microneutralization assays showed that the HA hexa-mutant reassortant virus (RG1) escaped A(H1N1)pdm09 hyper-immune ferret antiserum recognition. C57Black/6 mice that received the vaccine formulated with A/California/07/09 were challenged with 2×104 p.f.u. of either the 6 : 2 wild-type (WT) or RG1 viruses. Reductions in body weight loss, mortality rate and lung viral titre were observed in immunized animals challenged with the 6 : 2 WT virus compared to non-immunized mice. However, immunization did not protect mice challenged with RG1 virus. To further characterize the mutations causing this antigenic change, 11 additional RG viruses whose HA gene contained single or combinations of mutations were evaluated in vitro. Although the RG1 virus was still the least reactive against hyper-immune serum by HAI testing, mutations G158E and N159D within the Sa antigenic site appeared to play the major role in the altered antigenicity of the A(H1N1)pdm09 virus. These results show that the Sa antigenic site contains the most prominent epitopes susceptible to cause an antigenic drift, escaping actual vaccine protection.


p53 mutation does not correlate with radiosensitivity in 24 head and neck cancer cell lines.
The molecular basis of tumor response to therapeutic radiation is poorly understood. Recent evidence suggests the p53 tumor suppressor gene may be involved in production of the G1 arrest seen following DNA damage by X-irradiation. It has further been proposed that tumor cells lacking the p53 checkpoint function are likely to be more sensitive to cell killing by X-irradiation because these cells enter S phase despite unrepaired DNA damage. We tested the hypothesis that tumor cells with p53 mutations are more radiosensitive by correlating the in vitro surviving fraction at 2 Gy with the mutational status of 24 head and neck squamous cell cancer cell lines. p53 mutations were present in 15 of 24 (63%) of tumors; all were homozygous changes occurring within exons 5-9. The surviving fraction at 2 Gy for the group with mutations was 0.568 compared to 0.507 for tumors without mutations (P = 0.28, Mann-Whitney test). Furthermore, no association between radiosensitivity and mutational type, codon location, or predicted amino acid alteration was noted. Our data do not support the hypothesis that p53 gene alteration predisposes tumor cells to increased cell killing via radiation.


Geriatric Traumatic Brain Injury: Epidemiology, Outcomes, Knowledge Gaps, and Future Directions.
This review of the literature on traumatic brain injury (TBI) in older adults focuses on incident TBI sustained in older adulthood ("geriatric TBI") rather than on the separate, but related, topic of older adults with a history of earlier-life TBI. We describe the epidemiology of geriatric TBI, the impact of comorbidities and pre-injury function on TBI risk and outcomes, diagnostic testing, management issues, outcomes, and critical directions for future research. The highest incidence of TBI-related emergency department visits, hospitalizations, and deaths occur in older adults. Higher morbidity and mortality rates among older versus younger individuals with TBI may contribute to an assumption of futility about aggressive management of geriatric TBI. However, many older adults with TBI respond well to aggressive management and rehabilitation, suggesting that chronological age and TBI severity alone are inadequate prognostic markers. Yet there are few geriatric-specific TBI guidelines to assist with complex management decisions, and TBI prognostic models do not perform optimally in this population. Major barriers in management of geriatric TBI include under-representation of older adults in TBI research, lack of systematic measurement of pre-injury health that may be a better predictor of outcome and response to treatment than age and TBI severity alone, and lack of geriatric-specific TBI common data elements (CDEs). This review highlights the urgent need to develop more age-inclusive TBI research protocols, geriatric TBI CDEs, geriatric TBI prognostic models, and evidence-based geriatric TBI consensus management guidelines aimed at improving short- and long-term outcomes for the large and growing geriatric TBI population.


Long-term impact of a chronic disease management program on hospital utilization and cost in an Australian population with heart disease or diabetes.
To evaluate the longitudinal value of a chronic disease management program, My Health Guardian (MHG), in reducing hospital utilization and costs over 4 years. The MHG program provides individualized support via telephonic nurse outreach and online tools for self-management, behavior change and well-being. In follow up to an initial 18-month analysis of MHG, the current study evaluated program impact over 4 years. A matched-cohort analysis retrospectively compared MHG participants with heart disease or diabetes (treatment, N = 4,948) to non-participants (comparison, N = 28,520) on utilization rates (hospital admission, readmission, total bed days) and hospital claims cost savings. Outcomes were evaluated using regression analyses, controlling for remaining demographic, disease, and pre-program admissions or cost differences between the study groups. Over the 4 year period, program participation resulted in significant reductions in hospital admissions (-11.4%, P < 0.0001), readmissions (-36.7%, P < 0.0001), and bed days (-17.2%, P < 0.0001). The effect size increased over time for admissions and bed days. The relative odds of any admission and readmission over the 4 years were 27% and 45% lower, respectively, in the treatment group. Cumulative program savings from reduced hospital claims was $3,549 over 4-years; savings values for each program year were significant and increased with time (P = 0.003 to P < 0.0001). Savings calculations did not adjust for pooled costs (and savings) in Australia's risk equalization system for private insurers. Results confirm and extend prior program outcomes and support the longitudinal value of the MHG program in reducing hospital utilization and costs for individuals with heart disease or diabetes and demonstrate the increasing program effect with continued participation over time.


Dietary restriction increases insulin sensitivity and lowers blood glucose in rhesus monkeys.
Insulin sensitivity and glucose tolerance typically decline during later life. In a multidimensional randomized trial of the effects of dietary restriction started in adulthood on the processes of aging, we are studying insulin sensitivity and glucoregulation longitudinally in control (C, n = 15, fed a defined diet ad libitum for 6-8 h/day) and restricted (R, n = 15, fed 30% less than C) monkeys using the Modified Minimal Model method. Linear rates of change were calculated for individual animals through 30 mo of diet treatment and compared between treatment groups. Basal glucose, basal insulin, and insulin responses to glucose and tolbutamide increased for C and decreased for R animals (P < or = 0.002), whereas insulin sensitivity decreased for C and increased for R (P = 0.008). Glycosylated hemoglobin at 30 mo was marginally lower in R (P = 0.06) and was positively correlated with fasting plasma glucose (r = 0.508, P < 0.001). Insulin changes were significantly correlated with changes in adiposity (weight and abdominal circumference). Identification of the mechanisms through which these effects are achieved may aid in ameliorating glucose intolerance, insulin resistance, and associated illnesses in older persons.


Coping Strategies Mediate the Effect of Stressful Life Events on Schizotypal Traits and Psychotic Symptoms in 22q11.2 Deletion Syndrome.
Converging evidence suggests that psychosis emerges from the complex interaction of genetic and environmental factors. Stressful life events (SLEs) play a prominent role in combination with coping strategies and with a dysfunctional hypothalamus-pituitary-adrenal axis (HPAA). It has been proposed that the framework of schizotypy might help disentangle the interaction between genetic and environmental factors in the pathogenesis of psychosis. Similarly, 22q11.2 deletion syndrome (22q11DS) is considered as a genetic model of psychosis vulnerability. However, SLE and coping strategies remain largely unexplored in 22q11DS. Moreover, the HPAA has not been systematically investigated in this population. Here, we explored the correlation between SLE, emotional coping strategies, schizotypal personality traits, subthreshold psychotic symptoms in a sample of 43 healthy controls (HCs) compared with 59 individuals with 22q11DS. In the latter, we also explored the correlation with pituitary volume as estimated from structural magnetic resonance imaging. We found that SLE and negative coping strategies were correlated with schizotypal personality traits in both HCs and 22q11DS, and with psychotic symptoms in the 22q11DS group only, whereas reduced pituitary volume correlated with general psychopathology. Moreover, dysfunctional coping mediated the effect of SLE on schizotypal personality traits and psychotic symptoms in 22q11DS. Our findings recapitulate evidence in nonsyndromic patients and confirm the central role of stress and coping in the pathogenesis of psychosis. More broadly, they highlight the importance of environmental factors in the pathway to psychosis in 22q11DS, suggesting a strong rationale for the implementation of stress and particularly coping-oriented interventions in this population.


Concomitant presence of inflammatory fibroid polyp and carcinoma or adenoma in the stomach.
Four instances (8%) of inflammatory fibroid polyp (IFP) with concomitant adenocarcinoma or adenoma, in the same area, were noted among 50 cases of IFP of the stomach. Adding two cases from other sources, four cases of gastric IFP concomitant with an adenocarcinoma and two of gastric IFP concomitant with an adenoma were studied histopathologically and immunohistochemically. All lesions were located in the gastric antrum and they were restricted to the mucosa in three, and they involved both mucosa and submucosa in the other three. Neither S100 protein nor factor VIII-associated antigen was recognized in the principal component cells, using an immunoperoxidase technique. This finding suggests the conventional view that the proliferating cells were neither neurogenic nor angioblastic, but rather, were fibroblastic in origin. The four concomitant carcinomas were early adenocarcinomas restricted to within the mucosa, and the two concomitant adenomas were tubular adenomas with a moderate epithelial atypia. All these neoplasms were present in or adjacent to the IFP. We emphasize that the IFP, albeit benign, may carry an adenocarcinoma or an adenoma.


Customizable rigid head fixation for infants: technical note.
The need and advantages of rigid fixation of the head in cranial surgeries are well documented (Berryhill et al., Otolaryngol Head Neck Surg 121:269-273, 1999). Head fixation for neurosurgical procedures in infants and in early years has been a challenge and is fraught with risk. Despite the fact that pediatric pins are designed, rigid head fixation involving direct application of pins to the head of infants and slightly older children is still generally not safe (Agrawal and Steinbok, Childs Nerv Syst 22:1473-1474, 2006). Yet, there are some surgeries in which some form of rigid fixation is required (Agrawal and Steinbok, Childs Nerv Syst 22:1473-1474, 2006). We describe a simple technique to achieve rigid fixation of the head in infants for neurosurgical procedures. This involves applying a head band made of Plaster of Paris (POP) around the head and then applying the fixation pins of the fixation frame directly on to the POP. We have used this technique of head fixation successfully for infants with no complications.


Flavor-cyber-agriculture: Optimization of plant metabolites in an open-source control environment through surrogate modeling.
Food production in conventional agriculture faces numerous challenges such as reducing waste, meeting demand, maintaining flavor, and providing nutrition. Contained environments under artificial climate control, or cyber-agriculture, could in principle be used to meet many of these challenges. Through such environments, phenotypic expression of the plant-mass, edible yield, flavor, and nutrients-can be actuated through a "climate recipe," where light, water, nutrients, temperature, and other climate and ecological variables are optimized to achieve a desired result. This paper describes a method for doing this optimization for the desired result of flavor by combining cyber-agriculture, metabolomic phenotype (chemotype) measurements, and machine learning. In a pilot experiment, (1) environmental conditions, i.e. photoperiod and ultraviolet (UV) light (known to affect production of flavor-active molecules in edible plants) were applied under different regimes to basil plants (Ocimum basilicum) growing inside a hydroponic farm with an open-source design; (2) flavor-active volatile molecules were measured in each plant using gas chromatography-mass spectrometry (GC-MS); and (3) symbolic regression was used to construct a surrogate model of this chemistry from the input environmental variables, and this model was used to discover new combinations of photoperiod and UV light to increase this chemistry. These new combinations, or climate recipes, were then implemented in the hydroponic farm, and several of them resulted in a marked increase in volatiles over control. The process also led to two important insights: it demonstrated a "dilution effect", i.e. a negative correlation between weight and desirable chemical species, and it discovered the surprising effect that a 24-hour photoperiod of photosynthetic-active radiation, the equivalent of all-day light, induces the most flavor molecule production in basil. In this manner, surrogate optimization through machine learning can be used to discover effective recipes for cyber-agriculture that would be difficult and time-consuming to find using hand-designed experiments.


Functional effects of food components and the gastrointestinal system: chicory fructooligosaccharides.
Functional food science, as recently proposed by ILSI Europe, opens new perspectives in nutrition and food sciences. The systematic investigation of the interactions between food components or food ingredients and genomic, biochemical, cellular, or physiological functions is a unique way to improve both our knowledge and the role of nutrition in maintaining good health and in preventing disease. However, such basic knowledge is insufficient to justify claims, unless it is confirmed through relevant nutrition studies aimed at demonstrating the same effect and its positive consequences in humans. In the first stage, this demonstration will in most cases justify functional (physiological) claims (e.g., bifidogenic effect for fructooligosaccharides, bulking effect for nondigestible carbohydrates, protection against oxidative stress for antioxidants) with no reference to any health benefit. A true health claim will require, in most cases, additional studies involving large populations and long-term trials. It is anticipated that the better we understand the mechanism of interactions between food components and specific biological functions, the more we will be able to demonstrate functional effects, and the easier it will be to accumulate convincing evidence in favor of health promotion or disease prevention. Because of both its direct contact with eaten foods and the diversity of its functions, the GI system is a potential target for many functional effects. Until now, only a limited number of these effects have been investigated so as to justify functional claims. Improvement of glucose absorption (leading to physiological glycemia and insulinemia), modulation of GI transit time, fecal bulking, acidification of colonic content, and control of cholesterol bioavailability are all recognized effects of dietary fiber. Balanced colonic microflora and immunostimulation are attributed to the consumption of probiotics. Prebiotics selectively modify the colonic microbiota and modulate hepatic lipogenesis. According to the ILSI Europe strategy for the development of functional foods, all these effects are of interest. Their support by sound scientific arguments will be a necessary condition for their implementation in food science and nutrition for the benefit of human health.


Ligand-based target prediction with signature fingerprints.
When evaluating a potential drug candidate it is desirable to predict target interactions in silico prior to synthesis in order to assess, e.g., secondary pharmacology. This can be done by looking at known target binding profiles of similar compounds using chemical similarity searching. The purpose of this study was to construct and evaluate the performance of chemical fingerprints based on the molecular signature descriptor for performing target binding predictions. For the comparison we used the area under the receiver operating characteristics curve (AUC) complemented with net reclassification improvement (NRI). We created two open source signature fingerprints, a bit and a count version, and evaluated their performance compared to a set of established fingerprints with regards to predictions of binding targets using Tanimoto-based similarity searching on publicly available data sets extracted from ChEMBL. The results showed that the count version of the signature fingerprint performed on par with well-established fingerprints such as ECFP. The count version outperformed the bit version slightly; however, the count version is more complex and takes more computing time and memory to run so its usage should probably be evaluated on a case-by-case basis. The NRI based tests complemented the AUC based ones and showed signs of higher power.


Genome sequences of segmented filamentous bacteria in animals: implications for human research.
Experimental studies indicate that segmented filamentous bacteria (SFB) can recapitulate fundamental immune responses, particularly in reference to Th 17 and regulatory T-cell activity. The recent description of whole genomic DNA sequences of mouse and rat SFBs and the comparison between these sequences opens new important perspectives. In particular this knowledge allows to perform quali-quantitative studies in human beings regarding these yet unculturable anaerobic bacteria. The comparative analysis of rat and mouse SFB 16S rRNA sequences suggests the existence of highly conserved regions that could represent SFB-specific molecular targets potentially useful to develop PCR probes for study SFB in humans. These molecular tools may be of inestimable value to evaluate the possible role played by SFB in the immune system physiology, as well as its potential involvement in human chronic inflammatory and autoimmune diseases.


Paget-Schroetter syndrome associated with FV:Q506 and prothrombin 20210A--a case report.
Effort thrombosis of the axillary-subclavian vein (Paget-Schroetter syndrome) develops usually secondary to heavy arm exertion. An underlying chronic venous compressive anomaly at the thoracic outlet or intimal damage of the axillary vein following forceful hyperabduction, external rotation of the shoulder joint has been proposed to explain the pathophysiology of this thrombosis. This condition is usually not attributed to an underlying hypercoagulability such as deficiency of natural coagulation inhibitors. Here, the authors present a case with thrombosis of the axillary-subclavian vein following an effort, with factor V Leiden and prothrombin 20210A mutations. Both factor V Leiden and the genetic variant in the prothrombin gene have been shown to confer an increased risk for venous thrombosis. Although rare, effort thrombosis may develop in a patient with hereditary thrombophilia, so laboratory evaluation should include the common causes of thrombosis.


SECOND ATTACKS OF POLIOMYELITIS : AN EXPERIMENTAL STUDY.
1. It was found in forty rhesus monkeys that intracerebral, intraocular, intracutaneous, intraperitoneal, intraspinal, and neural inoculations of poliomyelitis virus produced no lesions in the olfactory bulbs despite the fact that the animals contracted pronounced paralyses. This indicated that the virus could be restricted to certain neuronal systems. 2. Similarly intranasal inoculation of seven animals produced no lesions in the ciliary ganglia. 3. Two monkeys convalescent from an intracutaneous and an intracerebral inoculation respectively had further paralyses after intranasal inoculation of heterologous virus. A third animal convalescent from an intranasal inoculation showed extension of lesions after intranasal and intraocular inoculation with heterologous virus. 4. Two spinal animals in which an attack of poliomyelitis was limited to an isolated segment of spinal cord, contracted typical paralyses in the previously uninvaded portions of the CNS following intranasal inoculation of homologous virus. 5. Four of six convalescent monkeys showed extension of lesions but no clinical signs after homologous virus inoculation through a previously un-invaded portal. 6. Four animals convalescent from a unilateral intranasal inoculation showed evidence of new invasion in the opposite olfactory bulb but no extension of paralyses following a second inoculation of homologous virus into the appropriate nostril. 7. Two animals had second attacks after heterologous second inoculations. The intranasal portal was employed for both exposures. 8. It thus seems apparent that in the rhesus monkey a second attack of poliomyelitis, whether paralytic or not, seems to depend upon the strains of virus used and the degree to which virus is disseminated through the neuraxis during the first exposure. 9. The above experimental data emphasize the difficulty of utilizing the rhesus monkey for experiments seeking to elucidate the mechanisms of immunity in man and suggest that human immunity to poliomyelitis does not result from immunization of the nervous system but rather is the result of some process which prevents infective quantities of active virus from reaching nervous tissue.


Pathophysiological classification of acquired bone marrow failure based on quantitative assessment of erythroid function.
Bone marrow failure encompasses a broad spectrum of disorders including aplastic, dysmyelopoietic and myelophtisic anemias. In the present study, these anemias were characterized according to the degree of erythroid proliferation and efficiency of erythropoiesis. Total erythropoietic activity was evaluated in 43 patients by measuring the erythron transferrin uptake (ETU). It averaged 20% of basal (range 3-43%) in 13 patients with severe aplastic anemia, 75% of basal (range 60-103%) in 3 patients with extensive bone marrow infiltration by neoplastic cells, 131% of basal (range 50-217%) in 16 patients with refractory anemia, and 452% of basal (range 63-720) in 11 patients with idiopathic refractory siderobastic anemia. Respective efficiencies of erythropoiesis were 74% in aplastic anemia, 70% with bone marrow infiltration, 46% in refractory anemia, and 14% in sideroblastic anemia. Based on the ETU, patients could be categorized into absolute marrow failure, relative marrow failure, and adequate erythropoietic response to anemia. This simple determination of proliferating activity of the erythroid marrow can provide useful information on the pathophysiology of marrow failure and a basis for the selection of therapeutic approaches.


Pathogenesis of experimental bovine respiratory syncytial virus infection in sheep.
Conventionally reared lambs, seronegative to bovine respiratory syncytial virus (RSV), were inoculated with bovine RSV by an aerosol route. A mild clinical response, characterized by transient fever at postinoculation day (PID) 4, occurred and was accompanied by serous oculonasal secretions in less than 50% of the infected lambs. A decrease in total WBC was observed on PID 6 and 11, with a reduction in the total numbers of lymphocytes. Bovine RSV was recovered from nasal secretions of inoculated lambs from PID 2 to 6. The lambs were euthanatized, and at necropsy, the virus was only isolated from lambs killed between PID 2 and 4. Serum neutralizing antibodies were noticed as early as PID 6 and peaked at PID 13 to 15. Neutralizing antibodies were also detected at low concentration in pulmonary washings. Viral antigen was mainly located in alveolar walls and occasionally in bronchiolar epithelium and alveolar macrophages in lambs killed between PID 2 to 4. Later, specific fluorescence was only present in alveolar walls and was not observed in tissue from lambs killed after PID 22. Budding and free virus-like particles were observed by electron microscopy in type I and type II alveolar epithelial cells. Outstanding macroscopic lesions were not seen. Histologically, mild bronchiolitis and alveolitis were observed.


Optimization of a biomimetic bone cement: role of DCPD.
We previously proposed a biomimetic α-tricalcium phosphate (α-TCP) bone cement where gelatin controls the transformation of α-TCP into calcium deficient hydroxyapatite (CDHA), leading to improved mechanical properties. In this study we investigated the setting and hardening processes of biomimetic cements containing increasing amounts of CaHPO(4)·2H2O (DCPD) (0, 2.5, 5, 10, 15 wt.%), with the aim to optimize composition. Both initial and final setting times increased significantly when DCPD content accounts for 10 wt.%, whereas cements containing 15 wt.% DCPD did not set at all. Differential scanning calorimetry (DSC), X-ray diffraction (XRD), thermogravimetry (TG) and scanning electron microscopy (SEM) investigations were performed on samples maintained in physiological solution for different times. DCPD dissolution starts soon after cement preparation, but the rate of transformation decreases on increasing DCPD initial content in the samples. The rate of α-TCP to CDHA conversion during hardening decreases on increasing DCPD initial content. Moreover, the presence of DCPD prevents gelatin release during hardening. The combined effects of gelatin and DCPD on the rate of CDHA formation and porosity lead to significantly improved mechanical properties, with the best composition displaying a compressive strength of 35 MPa and a Young modulus of 1600 MPa.


Quality assurance for in vitro alternative test methods: quality control issues in test kit production.
In vitro toxicology methods are being adopted by regulatory agencies worldwide. Many of these methods have been validated by using proprietary materials, often in the form of test kits. Guidelines for the use of Good Laboratory Practice methods for in vitro methods have been proposed. However, users of the data from these methods also need to be reassured that the proprietary materials and the test kits will provide consistent, good quality data over time, not just during the validation process. This paper presents an overview of the methods currently used by representatives of kit manufacturers and contract testing laboratories to ensure that the results from methods that utilise test kits are reproducible over time and across different types of test materials. This information will be valuable as a basis for future discussion on the need for formalised oversight of the quality of these materials.


PCR ribotyping and arbitrarily primed PCR for typing strains of Clostridium difficile from a Polish maternity hospital.
Detection of the source of Clostridium difficile strains is of importance for the control of the nosocomial spread of this microorganism. For this purpose, vaginal and rectal swabs from 183 mothers, duplicate fecal samples (taken on days 1 and 4 after birth) from 183 neonates, and 94 environmental samples were cultured for C. difficile. The microorganism was never detected in the meconium obtained on day 1 after birth. On the other hand, an incidence of 17% C. difficile positivity was noted in the fecal samples obtained on day 4 after birth. Forty-two percent of the 31 colonized neonates had been delivered with complications. The bacteria were never encountered in the rectal swabs of the mothers, and C. difficile was identified in only one vaginal swab. In contrast, 13% of the environmental samples were positive for C. difficile. No major difference was encountered between patient and environmental isolates with respect to toxigenicity (58 to 65% toxigenic isolates). All strains were subsequently typed by PCR amplification of the 16S-23S ribosomal intergenic spacer regions and by arbitrarily primed PCR (AP-PCR) with different primers and combinations thereof. All environmental isolates and 11 of 31 neonatal strains were of a single type. The vaginal strain was unique, and among the maternity ward- and neonate-related isolates, only two additional AP-PCR types were identified. When a collection of C. difficile strains from patients hospitalized in other institutions and suffering from antibiotic-associated diarrhea or pseudomembranous colitis was analyzed in a similar manner, it appeared that the strain from the maternity ward was unique. The other strain commonly encountered among the neonates was also identified frequently among the isolates from patients with antibiotic-associated diarrhea or pseudomembranous colitis, indicating its general occurrence. On the basis of both epidemiological studies and PCR-mediated genotyping, it was shown that the environment and not the birth canal is the major source of C. difficile acquisition by neonates in this maternity hospital setting. Furthermore, AP-PCR appears to be a fast and useful method for epidemiologically relevant typing of C. difficile isolates.


Facial electromyographic responses to emotional information from faces and voices in individuals with pervasive developmental disorder.
Despite extensive research, it is still debated whether impairments in social skills of individuals with pervasive developmental disorder (PDD) are related to specific deficits in the early processing of emotional information. We aimed to test both automatic processing of facial affect as well as the integration of auditory and visual emotion cues in individuals with PDD. In a group of high-functioning adult individuals with PDD and an age- and IQ-matched control group, we measured facial electromyography (EMG) following presentation of visual emotion stimuli (facial expressions) as well as the presentation of audiovisual emotion pairs (faces plus voices). This emotionally driven EMG activity is considered to be a direct correlate of automatic affect processing that is not under intentional control. Our data clearly indicate that among individuals with PDD facial EMG activity is heightened in response to happy and fearful faces, and intact in response to audiovisual affective information. This study provides evidence for enhanced sensitivity to facial cues at the level of reflex-like emotional responses in individuals with PDD. Furthermore, the findings argue against impairments in crossmodal affect processing at this level of perception. However, given how little comparative work has been done in the area of multisensory perception, there is certainly need for further exploration.


Factors associated with emergency department visits for migraine: an observational study.
This study aims to determine why patients with migraine present to an emergency department (ED). While migraine accounts for over 800,000 ED visits annually, no prospectively gathered data characterize patients' reasons for presenting to an ED. We prospectively interviewed 309 consecutive patients presenting to an urban ED for headache. Patients were asked 100 closed-ended questions regarding sociodemographics, headache history, and current headache attack. We performed descriptive analyses on patients fulfilling International Classification of Headache Disorders 2 migraine criteria. Of 186 patients who met migraine criteria, 77% (95% confidence interval [CI]: 71, 83%) had a primary care provider (PCP), 87% (95% CI: 82, 92%) had medical insurance, and 83% (95% CI: 77, 88%) had drug coverage. Fifty-three percent (95% CI: 46, 60%) reported that they previously visited a doctor for headache. Fifty-five percent (95% CI: 48, 62%) previously received a migraine diagnosis. Twenty-two percent (95% CI: 16, 28%) sought medical care for the current headache prior to ED presentation. Fifty-five percent (95% CI: 48, 63%) took abortive medication for migraine on the day of the ED visit. Median headache duration was 24 hours (IQR: 12-72). Forty-nine percent (95% CI: 42, 57%) screened positive for depression. The most common reason for visiting the ED was a perceived emergency condition or referral by a physician (33.3% [95% CI: 27, 40%]). Other commonly cited reasons related to access to care. Most migraineurs presenting to the ED have a PCP and health insurance. ED visits commonly result from an inability to access care elsewhere and because patients consider pain to be an emergency condition. Missed opportunities for diagnosis and treatment likely contribute to ED visits.


Impact of cholesterol on voids in phospholipid membranes.
Free volume pockets or voids are important to many biological processes in cell membranes. Free volume fluctuations are a prerequisite for diffusion of lipids and other macromolecules in lipid bilayers. Permeation of small solutes across a membrane, as well as diffusion of solutes in the membrane interior are further examples of phenomena where voids and their properties play a central role. Cholesterol has been suggested to change the structure and function of membranes by altering their free volume properties. We study the effect of cholesterol on the properties of voids in dipalmitoylphosphatidylcholine (DPPC) bilayers by means of atomistic molecular dynamics simulations. We find that an increasing cholesterol concentration reduces the total amount of free volume in a bilayer. The effect of cholesterol on individual voids is most prominent in the region where the steroid ring structures of cholesterol molecules are located. Here a growing cholesterol content reduces the number of voids, completely removing voids of the size of a cholesterol molecule. The voids also become more elongated. The broad orientational distribution of voids observed in pure DPPC is, with a 30% molar concentration of cholesterol, replaced by a distribution where orientation along the bilayer normal is favored. Our results suggest that instead of being uniformly distributed to the whole bilayer, these effects are localized to the close vicinity of cholesterol molecules.


The structural GDP/GTP cycle of Rab11 reveals a novel interface involved in the dynamics of recycling endosomes.
The small GTP-binding protein Rab11 is an essential regulator of the dynamics of recycling endosomes. Here we report the crystallographic analysis of the GDP/GTP cycle of human Rab11a, and a structure-based mutagenesis study that identifies a novel mutant phenotype. The crystal structures show that the nucleotide-sensitive switch 1 and 2 regions differ from those of other Rab proteins. In Rab11-GDP, they contribute to a close packed symmetrical dimer, which may associate to membranes in the cell and allow Rab11 to undergo GDP/GTP cycles without recycling to the cytosol. The structure of active Rab11 delineates a three-dimensional site that includes switch 1 and is separate from the site defined by the Rab3/Rabphilin interface. It is proposed to form a novel interface for a Rab11 partner compatible with the simultaneous binding of another partner at the Rabphilin interface. Mutation of Ser(29) to Phe in this epitope resulted in morphological modifications of the recycling compartment that are distinct from those induced by the classical dominant-negative and constitutively active Rab11 mutants. Recycling endosomes condensed in the perinuclear region where they retained recycling transferrin, and they clustered Rab11- and EEA1-positive membranes. Altogether, our study suggests that this mutation impairs a specific subset of Rab11 interactions, possibly those involved in cytoskeleton-based movements driving the slow recycling pathway.


Association of Dermatoglyphic Peculiarities with Dental Caries in Preschool Children of Lucknow, India.
Dermatoglyphics refers to study of the intricate dermal ridge configurations on the skin covering the palmar and plantar surfaces of hand and feet. The basis of considering dermatoglyphic patterns as genetic marker for dental caries is that the epithelium of finger buds as well as enamel has ectodermal origin, and both develop at the same time of intrauterine life. To assess the relationship between fingerprint patterns and dental caries among preschool children of Lucknow city. This study was of cross-sectional design. The study group comprised 512 preschool children 2-6 years of age. The prevalence of caries was recorded using "Dentition status and treatment needs" (WHO basic oral health assessment form, 1997). They were divided into three groups as follows: Group I (dmft score = 0-2), group II (dmft score = 3-4) and group III (dmft score ≥5). The handprints of each child were taken using a stamp pad. The fingertip patterns were analyzed according to the classical method and were classified according to the topological method. The frequency of occurrence of type of dermatoglyphic pattern on fingertip of each digit was noted. Chi-square test was used to test the significant difference in proportions. Means were compared using Student's t-test and analysis of variance (ANOVA) or F-test. Subjects belonging to groups II and III showed maximum occurrence of whorl pattern on all digits. Group I subjects had maximum occurrence of arch pattern. All the variables had statistically significant value, with a degree of divergence of specific dermatoglyphic patterns among all three groups. The dental caries susceptibility of an individual increased with incidence of whorl pattern and it decreased with incidence of arch pattern. How to cite this article: Singh E, Saha S, Jagannath GV, Singh S, Saha S, Garg N. Association of Dermatoglyphic Peculiarities with Dental Caries in Preschool Children of Lucknow, India. Int J Clin Pediatr Dent 2016;9(1):39-44.


High level of galectin-1 expression is a negative prognostic predictor of recurrence in laryngeal squamous cell carcinomas.
Monitoring of gene-expression profiles is assumed to refine tumor characterization of laryngeal squamous cell carcinomas (LSCCs) with a therapeutic perspective. This is especially expected for adhesion/growth-regulatory effectors such as galectins, a class of endogenous lectins. Using computer-assisted microscopy, we investigated the prognostic value contributed by the quantitative determination of the immunohistochemical levels of expression of galectin-1, -3 and -7 in a series of 62 LSCCs including 42 low- and 20 high-stage LSCCs. As galectin-1 may have a key role leading to a tumor escape from immune surveillance, we also investigated whether or not the level of galectin-1 expression correlated with lymphocyte infiltration in LSCCs. The immunohistochemical determination of expression of galectin-1 is of prognostic value in human squamous laryngeal cancers. LSCCs that display high levels of galectin-1 have worse prognoses than laryngeal cancers with low levels of galectin-1 expression. Elevation of galectin-1 levels in laryngeal cancers can contribute to the process of tumor immune escape by killing the activated T-cells and other protumoral activities such as promoting motility or activity of oncogenic H-Ras proteins. The quantitative determination of galectin-1 in LSCCs is an independent prognostic marker when opposed to TNM staging. It has the potential to identify patients unlikely to benefit from T-cell-mediated immunotherapy, although the definitive effector function from its pro- and antitumoral activity profile has not been delineated.


Smoking prevention that increases tax revenue: tobacco taxes in the Czech Republic 2004-2020.
Increasing tobacco excise tax is one of the most effective smoking-prevention tools. The aim of the study is to assess the use of this tool in the Czech Republic (CR) by studying trends in cigarette taxes, prices, tax revenue, and the affordability of cigarettes in the CR. Data on cigarette consumption, their tax rates, price, and tax revenue in the CR for 2004-2020 come from multiple sources. We used the consumer price index to convert nominal values to real values. Given an average daily consumption of 12.7 cigarettes per smoker, the affordability of cigarettes was measured as a percentage of the average monthly wage needed to buy 19 cigarette packs. Despite recent increases in excise taxes, cigarettes in the CR are becoming more affordable. We found that the affordability of cigarettes was greater in 2020 than in 2008. The values of both the specific and the minimum excise taxes are currently being eroded by inflation. Cigarette consumption has declined from 2015 to 2020, and the government still received a bit more excise tax revenue due to its earlier tax policy. However, if taxes are not increased further, the revenue will start to decline. Despite the trend of increasing tobacco taxes in the CR, both the affordability of cigarettes and their use are still high in the country. This means that the CR is not using tax policy effectively enough to reduce smoking prevalence. It needs a substantial and sudden tax increase, in addition to the currently planned tax increases, to reduce smoking prevalence and lower the burden of tobacco use in the economy. Such a move would not only improve public health in the CR, but also increase government revenue.


Amniotic fluid exhibits an innate inhibitory activity against HIV type 1 replication in vitro.
Indirect evidence suggests that amniotic fluid (AF) may play a role in the pathogenesis of in utero HIV-1 transmission. The purpose of this study was to evaluate the potential innate inhibitory role of AF on HIV replication, which may contribute to protection of the fetus against intrauterine transmission. AF was collected from term HIV-1-negative women undergoing scheduled cesarean section. The inhibitory effect of AF against HIV-1(BA-L) replication was tested in vitro with or without the addition of protease inhibitor cocktail (PIC) in PHA-stimulated PBMC cultures. Quantitative measurement of human neutrophil peptides 1-3 (HNP1-3) was performed on all AF samples, using an ELISA assay. AF exhibited a dose-dependent inhibitory activity against HIV-1(BA-L) replication, with all samples (n=12) reaching significant inhibitory effect using 50% AF. In vitro, this activity decreased over time, but was able to be sustained with the addition of PIC. The HNP1-3 concentration in AF samples (n=12) ranged from undetectable (<41 pg/ml, n=3) to >250,000 pg/ml with a median of 5,146 pg/ml. AF exhibited a significant and dose-dependent innate inhibitory activity against HIV-1 replication, which was present in all AF samples tested. This effect was prolonged in the presence of PIC, suggesting that the inhibitory factor was in the cell-free protein fraction. The HNP1-3 concentration in AF was in the subinhibitory range for HIV with no correlation between its concentration and the HIV-1 inhibitory activity. These data show the presence of a significant innate inhibitory activity against HIV in AF.


Ethanol effects on volitional versus reflexive saccades.
The effects of a moderate alcohol dose on reflexive and volitional saccades were compared. Healthy male subjects (n = 17) made horizontal reflexive saccades, antisaccades and self-paced saccades before and after the ingestion of a body-weight-related dose of alcohol. At a mean blood alcohol concentration of 0.044%, reflexive saccade latency significantly increased although accuracy remained unimpaired at each target amplitude. No significant ethanol effects were noted for antisaccade accuracy and latency. Antisaccade error rates decreased significantly, likely demonstrating a learning effect. The mean rate of self-paced saccades did not significantly change post-ethanol and their accuracy was also unimpaired. All saccades share a final common pathway for their execution. It thus seems reasonable to propose that ethanol modulated connections between the posterior parietal cortex and superior colliculus to increase reflexive saccade latency. Higher-order paradigms, which require inhibition and are mediated by frontal areas, might be impaired at greater blood alcohol concentrations.


Tumor-associated macrophage/microglia infiltration in human gliomas is correlated with MCP-3, but not MCP-1.
The monocyte chemotactic protein 3 (MCP-3) belongs to the MCP subgroup of the CC chemokines and promotes chemotaxis of immune cells. MCP-1 is believed to play an important role in monocyte infiltration into tumor tissues; however, the relationship between tumor-infiltrating macrophage/microglia (TIM/M) and the expression of chemokines has not been investigated in detail in human glioma samples; therefore, we first examined the expression of several chemokines and chemokine receptors in human tumor cell lines, which included glioma lines, using real-time PCR. We found that several glioma lines expressed MCP-3 predominantly, and not MCP-1. In order to assess the significance of MCP-3 expression in human glioma tissues, we then examined the number of CD68+ TIM/M, the percentage of TIM/M in the total cell population, and the expression of MCP-1 and MCP-3 in glioma tissues. There was a correlation between the percentage of TIM/M and MCP-3 expression levels; however, there was no correlation between the percentage of TIM/M and MCP-1 expression. There was no correlation between the number of TIM/M and prognosis of patients. These data indicate that tumor cell-derived MCP-3, but not MCP-1, facilitates the infiltration of macrophage/microglia into tumor tissues. This is the first study that clearly compared the significance of MCP-3 with that of MCP-1 in the tumor infiltration rates of TIM/M.


Outcome of Targeted vs Empiric Antibiotic Therapy in the Treatment of Spondylodiscitis: A Retrospective Analysis of 201 Patients.
Antibiotic treatment of spondylodiscitis is influenced by antibiotic stewardship; specifically, empirical antibiotic therapy is avoided in favor of-delayed-targeted antibiotic therapy after microbiological diagnosis. Only patients with neurological deficits or clinical signs of sepsis should be treated by empirical antibiotic therapy. However, the level of evidence for this treatment concept is weak. The aim of this study was to investigate whether patients who were treated with a nontargeted antibiotic therapy show a worse outcome than patients who were exclusively treated with targeted antibiotic therapy. A retrospective single-center data analysis. We included 201 patients with spondylodiscitis who were treated at the authors' institution between 2013 and 2020. Mortality rate, time in hospital, development of laboratory parameters, and development of pain (visual analog scale). We performed a retrospective data analysis of patients who were treated for spondylodiscitis from January 2013 to March 2020. Clinical and demographic data as well as outcome and complications were recorded and analyzed. We investigated whether patients who were treated by a nontargeted antibiotic therapy had a worse outcome than patients who were exclusively treated by targeted antibiotic therapy and which other clinical factors had an impact on clinical outcome. A total of 201 patients were included, 37 of whom developed sepsis during their hospital stay, and 14 of these 37 patients died. The 14 patients who died represented (10.0%) of the subgroup who received an empirical antibiotic treatment before the targeted antibiotic therapy (n = 141). There were 0 deaths in the subgroup who did not receive an empirical antibiotic treatment (P < 0.05). The time to diagnosis since admission to the hospital was 10.5 (2.2) days in patients with sepsis; in patients without sepsis, it was 5.2 (0.9) days (P < 0.05). Based on the results of this investigation, the authors recommend delaying antibiotic therapy until targeted antibiotic therapy is possible. Furthermore, early diagnosis of spondylodiscitis and prevention and early detection of sepsis are essential to reduce the mortality rate of patients with spondylodiscitis.


CSF-1R regulates non-small cell lung cancer cells dissemination through Wnt3a signaling.
Therapeutic antibodies targeting colony stimulating factor 1 receptor (CSF-1R) to block colony stimulating factor-1/colony stimulating factor 1 receptor (CSF-1/CSF-R) signaling axis have exhibit remarkable efficacy in the treatment of malignant tumor. Yet, little is known about the effects of intrinsic CSF-1R in human non-small-cell carcinoma (NSCLC). Here we demonstrated that NSCLC cell-intrinsic CSF-1R promoted cells growth and metastasis both in vitro and in vivo. CSF-1R knocked-down by transfecting with shRNA target CSF-1R suppressed NSCLC cells proliferation and tumor growth in nude mice. Conversely, ectopic expression of CSF-1R promoted cells proliferation and accelerated tumor growth. Mechanistically, the NSCLC CSF-1R modulated downstream effectors of phosphatidylinositol 3-kinase (PI3K) signaling. In addition, CSF-1R overexpression significantly enhanced NSCLC cells mobility, invasion and epithelial-mesenchymal transition (EMT) process, whereas silencing CSF-1R inhibits these phenotypes. Microarray analysis suggested that Wnt family member 3a (Wnt3a) function as a downstream factor of CSF-1R. On account of this, we future identified CSF-1R/Wnt3a a signaling pathway sustained NSCLC cells metastasis. Finally, in patients, CSF-1R and Wnt3a expression positively correlated with the of NSCLC patients. Our results identify NSCLC cell intrinsic functions of CSF-1R/Wnt3a axis in dissemination of NSCLC.


Methanol carbonylation to acetaldehyde on Au particles supported by single-layer MoS2grown on silica.
Homogenous single-layer MoS2films coated with sub-single layer amounts of gold are found to isolate the reaction of methanol with carbon monoxide, the fundamental step toward higher alcohols, from an array of possible surface reactions. Active surfaces were prepared from homogenous single-layer MoS2films coated with sub-single layer amounts of gold. These gold atoms formed clusters on the MoS2surface. A gas mixture of carbon monoxide (CO) and methanol (CH3OH) was partially converted to acetaldehyde (CH3CHO) under mild process conditions (308 kPa and 393 K). This carbonylation of methanol to a C2species is a critical step toward the formation of higher alcohols. Density functional theory modeling of critical steps of the catalytic process identify a viable reaction pathway. Imaging and spectroscopic methods revealed that the single layer of MoS2facilitated formation of nanoscale gold islands, which appear to sinter through Ostwald ripening. The formation of acetaldehyde by the catalytic carbonylation of methanol over supported gold clusters is an important step toward realizing controlled production of useful molecules from low carbon-count precursors.


Toward a definition of quality.
In a mail questionnaire to members of the American College of Physician Executives, physician executives were asked to rank and weight the importance of nine elements of a quality definition for each of six clinical scenarios. The scenarios differed in terms of the severity of illness on admission and in terms of outcome. The study suggests that evaluations of quality that do not assess severity of illness may be misleading. The study also demonstrates that some agreement on the relative importance of indicators of quality can be reached.


First Report of Bacterial Leaf Streak of Strelitzia reginae Caused by Burkholderia cepacia.
In 2011, the bacterial leaf streak disease of the monocotyledonous flowering plant, commonly known as bird of paradise (Strelitzia reginae), occurred in a nursery in Guangzhou, Guangdong Province, China. Lesions on diseased leaves began as water-soaked leaf spots or streaks near the central and secondary veins, eventually expanded along veins and became brown necrotic streaks. Occasionally, during wet conditions, seedlings were completely blighted. The disease incidence was about 12% in the nursery. Bacteria were consistently isolated on nutrient agar (NA) (4) from surface-sterilized symptomatic lesions and purified on NA. Three bacterial strains were tested for pathogenicity on S. reginae plants. Three plants were inoculated per bacterial strain (bacterial suspensions 107 CFU/ml in nutrient broth [NB] [4]) by wounding three young, fully expanded leaves (four wounds per leaf) with needle. Plants were placed in polyethylene bags 1 day before inoculation and maintained for 7 days after inoculation. Three control plants were inoculated with NB. Water-soaked areas on leaves were observed on all inoculated plants 7 days after inoculation. Within 10 days, brown streaks were observed. All strains induced similar symptoms as those observed on the plants in the nursery. Control plants showed no symptoms. For molecular identification, a near full-length sequence of the 16S rRNA gene was amplified from strain TNT1-1 (GenBank Accession No. JX901049.1) with primers 27F and 1492R (3), obtaining a PCR product of ~1,500 bp. A BLAST search in GenBank revealed the highest similarity (99.5%) to sequences of Burkholderia cepacia (FN178432.1 and FN178432.1). BIOLOG identification showed that TTN1-1 had the highest probability index of 0.85 and highest similarity index of 0.85 to B. cepacia. For biochemical characteristics, the strain was gram negative, anaerobic growth test negative, oxidase negative, catalase positive, did not produce fluorescent pigment on KB (4), did not grow on DIM agar (4), arginine dihydrolysis negative, nitrate reduction negative, starch hydrolysis negative, gelatin liquefaction negative, citrate, D-arabinose, L-fructose, trehalose, and maltose utilization positive, didn't produce acid from glucose, and grew on Tween 80 medium at 41°C. The above characteristics were identical to that of reference isolate B. cepacia ATCC 25416. Additionally, bacteria isolated on NA from the leading edge of lesions of inoculated plants with the strain were identical to the inoculated strain based on 16S rDNA sequence analysis, but no bacteria were recovered from the wounded sites on the control plants. Therefore, bacterial leaf streak of bird of paradise is caused by B. cepacia based on Koch's postulates. In contrast, two bacterial diseases on S. reginae were previously reported to be caused by Xanthomonas campestris (1) and B. gladioli (2) in the United States and Italy, respectively. A similar leaf streak disease on S. nicolai was caused by Acidovorax avenae subsp. avenae in the United States (5). To our knowledge, this is the first report of a leaf streak disease on S. reginae caused by B. cepacia. References: (1) A. R. Chase and J. B. Jones. Plant Dis. 71:845, 1987. (2) G. Cirvilleri et al. Plant Dis. 90:1553, 2006. (3) I. M. Lee et al. Appl. Environ. Microbiol. 63:2631, 1997. (4) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2001. (5) T. E. Seijo and N. A. Peres. Plant Dis. 95:1474, 2011.


Hookah smoking: behaviors and beliefs among young consumers in the United States.
Hookah smoking is growing in popularity among college-age students in the United States. Community-based, semistructured, in-depth interviews were conducted with 20 college-age individuals who had used a hookah to smoke tobacco. A structured analysis of responses was conducted and framed in terms of the health belief model's four main constructs. Although hookah smokers were aware of the potential severity of tobacco-related illnesses, they did not express a perceived susceptibility to those illnesses. To the extent that cues to action exist with respect to hookah smoking, they generally serve to promote a risky behavior that is perceived to be largely social in nature.


Multiple supramolecular structures formed by interaction of actin with protamine.
When protamine is added to actin, different supramolecular structures are formed depending on the molar ratio of the two proteins and of the ionic strength of the medium. At low ionic strength, and going from a molar ratio of protamine to G-actin of 4:1, 2:1 and 1:1, globular aggregates are first converted into extended structures and then to long threads in which the constituent ATP-G-actin is rapidly exchangeable with the actin of the medium. At high ionic strength {Tyrode [(1910) Arch. Int. Pharmacodyn. Ther.20, 205-212] solution}, starting from G-actin and protamine in the 1:1 molar ratio, long ropes are formed that can be resolved into intertwining filaments of 4-5nm diameter. The addition of protamine in a 1:1 molar ratio to a solution of F-actin in Tyrode solution causes the breakage of the actin filaments, which is also revealed by the decrease of the viscosity of the solution and the formation of ordered latero-lateral aggregates. The structures formed by reaction of protamine with G-actin can be separated from free G-actin and protamine by filtration through 0.45mum-pore-size Millipore filters. This technique has been exploited to study the exchange reaction between free actin and the actin-protamine complexes. For these studies the 1:1 actin-protamine complex formed at low ionic strength and the 2:1 actin-protamine complex formed in the presence of 23nm-free Mg(2+) have been selected. In the first case the exchange reaction is practically complete in the dead time of the experiment (20s). In the second case, where the complex operates like a true ATPase, the rate of the exchange is initially comparable with the rate of the ATP cleavage. Later on, however, the complex undergoes a change and the rate of the exchange between free actin and the actin bound to protamine becomes lower than the rate of the ATPase reaction. It is proposed that the ATP exchanges for ADP directly on the G-actin bound in the complex.


Protective effect of kombucha tea against tertiary butyl hydroperoxide induced cytotoxicity and cell death in murine hepatocytes.
Kombucha (KT), a fermented black tea (BT), is known to have many beneficial properties. In the present study, antioxidant property of KT has been investigated against tertiary butyl hydroperoxide (TBHP) induced cytotoxicity using murine hepatocytes. TBHP, a reactive oxygen species inducer, causes oxidative stress resulting in organ pathophysiology. Exposure to TBHP caused a reduction in cell viability, increased membrane leakage and disturbed the intra-cellular antioxidant machineries in hepatocytes. TBHP exposure disrupted mitochondrial membrane potential and induced apoptosis as evidenced by flow cytometric analyses. KT treatment, however, counteracted the changes in mitochondrial membrane potential and prevented apoptotic cell death of the hepatocytes. BT treatment also reverted TBHP induced hepatotoxicity, however KT was found to be more efficient. This may be due to the formation of antioxidant molecules like D-saccharic acid-1,4-lactone (DSL) during fermentation process and are absent in BT. Moreover, the radical scavenging activities of KT were found to be higher than BT. Results of the study showed that KT has the potential to ameliorate TBHP induced oxidative insult and cell death in murine hepatocytes more effectively than BT.


Red-Shifted FRET Biosensors for High-Throughput Fluorescence Lifetime Screening.
We have developed fluorescence resonance energy transfer (FRET) biosensors with red-shifted fluorescent proteins (FP), yielding improved characteristics for time-resolved (lifetime) fluorescence measurements. In comparison to biosensors with green and red FRET pairs (GFP/RFP), FPs that emit at longer wavelengths (orange and maroon, OFP/MFP) increased the FRET efficiency, dynamic range, and signal-to-background of high-throughput screening (HTS). OFP and MFP were fused to specific sites on the human cardiac calcium pump (SERCA2a) for detection of structural changes due to small-molecule effectors. When coupled with a recently improved HTS fluorescence lifetime microplate reader, this red-shifted FRET biosensor enabled high-precision nanosecond-resolved fluorescence decay measurements from microliter sample volumes at three minute read times per 1536-well-plate. Pilot screens with a library of small-molecules demonstrate that the OFP/MFP FRET sensor substantially improves HTS assay quality. These high-content FRET methods detect minute FRET changes with high precision, as needed to elucidate novel structural mechanisms from small-molecule or peptide regulators discovered through our ongoing HTS efforts. FRET sensors that emit at longer wavelengths are highly attractive to the FRET biosensor community for drug discovery and structural interrogation of new therapeutic targets.


KDmarkers: A biomarker database for investigating epigenetic methylation and gene expression levels in Kawasaki disease.
Kawasaki disease (KD) is a form of acute systemic vasculitis that primarily affects children and has become the most common cause of acquired heart disease. While the etiopathogenesis of KD remains unknown, the diagnostic criteria of KD have been well established. Nevertheless, the diagnosis of KD is currently based on subjective clinical symptoms, and no molecular biomarker is yet available. We have previously performed and combined methylation array (Illumina HumanMethylation450 BeadChip) and transcriptome array (Affymetrix GeneChip Human Transcriptome Array 2.0) to identify genes that are differentially methylated/expressed in KD patients compared with control subjects. We have found that decreased methylation levels combined with elevated gene expression can indicate genes (e.g., toll-like receptors and CD177) involved in the disease mechanisms of KD. In this study, we constructed a database called KDmarkers to allow researchers to access these valuable potential KD biomarkers identified via methylation array and transcriptome array. KDmarkers provides three search modes. First, users can search genes differentially methylated and/or differentially expressed in KD patients compared with control subjects. Second, users can check the KD patient groups in which a given gene is differentially methylated and/or differentially expressed. Third, users can explore the DNA methylation levels and gene expression levels in all samples (KD patients and controls) for a particular gene of interest. We further demonstrated that the results in KDmarkers are strongly associated with KD immune responses. All analysis results can be downloaded for downstream experimental designs. KDmarkers is available online at https://cosbi.ee.ncku.edu.tw/KDmarkers/.


Invasive salmonids and lake order interact in the decline of puye grande Galaxias platei in western Patagonia lakes.
Salmonid fishes, native to the northern hemisphere, have become naturalized in many austral countries and appear linked to the decline of native fishes, particularly galaxiids. However, a lack of baseline information and the potential for confounding anthropogenic stressors have led to uncertainty regarding the association between salmonid invasions and galaxiid declines, especially in lakes, as these have been much less studied than streams. We surveyed 25 lakes in the Aysén region of Chilean Patagonia, including both uninvaded and salmonid-invaded lakes. Abundance indices (AI) of Galaxias platei and salmonids (Salmo trutta and Oncorhynchus mykiss) were calculated using capture-per-unit-effort data from gillnets, minnow traps, and electrofishing. We also measured additional environmental variables, including deforestation, lake morphometrics, altitude, and hydrological position (i.e., lake order). An information-theoretic approach to explaining the AI of G. platei revealed that by far the strongest effect was a negative association with the AI of salmonids. Lake order was also important, and using structural equation modeling, we show that this is an indirect effect naturally constraining the salmonid invasion success in Patagonia. Supporting this conclusion, an analysis of an independent data set from 106 mountain lakes in western Canada showed that introduced salmonids are indeed less successful in low-order lakes. Reproductive failure due to insufficient spawning habitat and harsh environmental conditions could be the cause of these limits to salmonid success. The existence of this effect in Chilean Patagonia suggests that low-order lakes are likely to provide natural ecological refugia for G. platei. Finally, pristine, high-order lakes should be actively protected as these have become rare and irreplaceable unspoiled references of the most diverse, natural lake ecosystems in Patagonia.


On the molecular mechanism of the circadian clock. The 41,000 M(r) clock protein of Chlorella was identified as 3-phosphoglycerate kinase.
A 41,000 M(r) polypeptide of Chlorella exhibits a circadian rhythm in its synthesis and possesses characteristic features of a putative essential clock protein as was proposed by the coupled translation-membrane model. Purification of this polypeptide and a microsequencing analysis yielded a N-terminal sequence of 35 amino acids that showed no homology to known sequences that were thought to be involved in circadian rhythm such as the per gene of Drosophila and the frq gene of Neurospora. However, strong homology was observed to 3-phosphoglycerate kinase (PGK) of different organisms. The highest homology (83%) of this Chlorella sequence was found with the PGK of wheat chloroplast. PGK activity and the 41,000 M(r) polypeptide co-purified through differential centrifugation and gel filtration. These data, and comparison with the physical properties of other known PGK molecules, support the conclusion that the 41,000 M(r) polypeptide of Chlorella, a candidate for a putative essential clock protein, is 3-phosphoglycerate kinase.


Pertussis: an old disease that is still with us.
Our knowledge of pathogenesis, clinical presentation and prevention of pertussis has improved substantially over recent years. We now better understand the function of long-known virulence factors for Bordetella pertussis, and genome sequencing has identified a multitude of new proteins; their functions are yet to be elucidated. Furthermore, improved diagnostic tools have revealed the broad spectrum of disease, and new insights into the host's immune response have been gained. Finally, the development, evaluation, licensing and implementation of several new acellular pertussis vaccines with high acceptance have changed the epidemiology of pertussis in many countries (i.e. a shift towards an increasing burden of disease in adolescent persons and adults). These developments are likely to have great impact on the daily practice not only of paediatricians, but also of general practitioners, internists, gynaecologists and many other specialists in adolescent and adult medicine. The present review provides an update on recent progress.


Interphase cohesin regulation ensures mitotic fidelity after genome reduplication.
To ensure faithful genome propagation, mitotic cells alternate one round of chromosome duplication with one round of chromosome separation. Chromosome separation failure thus causes genome reduplication, which alters mitotic chromosome structure. Such structural alterations are well documented to impair mitotic fidelity following aberrant genome reduplication, including in diseased states. In contrast, we recently showed that naturally occurring genome reduplication does not alter mitotic chromosome structure in Drosophila papillar cells. Our discovery raised the question of how a cell undergoing genome reduplication might regulate chromosome structure to prevent mitotic errors. Here, we show that papillar cells ensure mitotic fidelity through interphase cohesin regulation. We demonstrate a requirement for cohesins during programmed rounds of papillar genome reduplication known as endocycles. This interphase cohesin regulation relies on cohesin release but not cohesin cleavage and depends on the conserved cohesin regulator Pds5 . Our data suggest that a distinct form of interphase cohesin regulation ensures mitotic fidelity after genome reduplication.