Human cerebral malaria: association with erythrocyte rosetting and lack of anti-rosetting antibodies.
Plasmodium falciparum isolates from 24 Gambian children with cerebral malaria and 57 children with mild forms of the disease were assessed for their ability to form erythrocyte rosettes. All isolates from the children with cerebral malaria were able to form rosettes, whereas those from children with mild forms of the disease did not form rosettes, or had a significantly lower rosetting rate. Plasma of children with cerebral malaria lacked anti-rosetting activity, whereas plasma of children with mild disease could often disrupt rosettes in vitro. A monoclonal antibody to P falciparum histidine rich protein (PfHRP1/KP/KAHRP) disrupted rosettes of many of the isolates in vitro indicating that the rosetting ligand is relatively conserved compared with ligands associated with endothelial cytoadherence. The findings strongly support the hypothesis that erythrocyte rosetting contributes to the pathogenesis of cerebral malaria and suggest that anti-rosetting antibodies protect against cerebral disease.


Differences between the Cerec-3D software versions 1000 and 1500.
The Cerec-3D software was introduced officially at the IDS 2003 with the version 2.00 R1000. The decisive difference compared with the preceding software generations was that the prepared tooth and the restoration were displayed three dimensionally. Thus, the virtual changes which could be made with the aid of editing tools could also be controlled and seen immediately in 3D. The latest officially available version of the Cerec-3D software version 2.10 R1500 differs in some further developments from the first version 2.00 R1000; these are for example: --If the semiautomatic margin detector is switched off while marking the preparation margin, the alternative intensity image appears instead of the 3D display. --Designing has been decisively improved by the extension of the "Drop" tool (function of continuous application and removal of material). --Under morphologically difficult circumstances, the design proposal can be magnified/reduced in size and repositioned in all spatial directions. --The Parameter setting again permits setting an adhesive gap width specific to the dentist and unit. As an example, the version 1500 software is demonstrated on a simple clinical case in which the occlusion points were defined virtually with the aid of a functional bite registration, eliminating the need for occlusal grinding-in.


The host-parasite relationship in pregnant cattle infected with Neospora caninum.
The protozoan parasite Neospora caninum is an important cause of reproductive disease in cattle worldwide. The dog is a definitive host for the parasite and the oocyst stage, shed in the faeces, is a source of infection for cattle through consumption of contaminated feed or water. In addition, transplacental transmission of N. caninum is a very efficient means of the parasite infecting a new host and this can occur in successive pregnancies and over several generations. Neospora parasites may cause disease during pregnancy resulting in death of the foetus or birth of live congenitally infected calves that may show some neurological clinical signs at birth. The stage of pregnancy at which infection/parasitaemia occurs is an important factor in determining disease severity. Neospora infection in the first trimester of pregnancy may have more severe consequences for the foetus compared with infection occurring in the final trimester. The host-parasite relationship during pregnancy is a fascinating interaction and research in this area will improve understanding of disease pathogenesis and the various consequences of the host immune response, being host-protective, parasite protective and contributing to disease pathology. Pregnancy poses an interesting problem for the immune system of the dam as she is essentially carrying a semi-allogeneic tissue graft (the foetus) without immunological rejection taking place. To facilitate the pregnancy the cytokine environment in the placenta favours the regulatory Th-2-type cytokines, whose role is to counteract the pro-inflammatory Th1-type immune responses. Protective immunity to N. caninum, similar to many other intracellular parasites, involves Th1-type immune responses, which may pose problems for the dam trying to control a Neospora infection during pregnancy. This paper will discuss the host-parasite relationship at different stages of gestation in pregnant cattle and review the implications of this research for our understanding of disease pathogenesis, parasite transmission and host protection.


Responsiveness of an activity tracker as a measurement tool in a knee osteoarthritis clinical trial (ACTIVe-OA study).
In osteoarthritis (OA) clinical trials, reliable and responsive outcome measures to document physical and functional improvements are limited. This study aimed to assess whether the use of an activity tracker in an OA clinical trial is a responsive measurement tool. Secondary objectives assessed feasibility and validity. We recruited 65 participants in a prospective cohort study nested in a placebo-controlled clinical trial of platelet-rich plasma injection in knee OA. Participants wore an activity tracker (Fitbit Flex 2), and a smartphone was preloaded with a mobile application (OApp) designed to monitor load rates as a surrogate of knee loading. Participants used the systems for 7 days at baseline and for 7 days before the 2-month follow-up assessment. Effect size (ES) and standardised response mean (SRM) were calculated for change in step count and knee loading rate and regularly used knee OA outcome measures. Correlation coefficients (r) were calculated to examine the strength of the association between outcome measures. . Step count showed a trivial ES and SRM and mean knee loading rate measurements a moderate ES and SRM. We found a weak but significant correlation between change in mean steps per day and global improvement overall (r= 0.28) and Western Ontario and McMaster Universities Osteoarthritis Index function (r = -0.28). Compliance was high with the activity trackers. Despite good feasibility, this study did not show significant responsiveness or validity of the activity trackers as compared with currently recommended outcome measures in OA clinical trials. The main challenge is the lack of a gold standard outcome measure to validate against, and because of the complex interplay between pain and measured function, a lack of correlation does not necessarily represent a failed validation in this context. ACTRN12617000853347. This trial is a substudy of the "Platelet-rich plasma as a symptom-and disease-modifying treatment for knee osteoarthritis - the RESTORE trial".


Fall armyworm (Spodoptera frugiperda Smith) feeding elicits differential defense responses in upland and lowland switchgrass.
Switchgrass (Panicum virgatum L.) is a low input, high biomass perennial grass being developed for the bioenergy sector. Upland and lowland cultivars can differ in their responses to insect herbivory. Fall armyworm [FAW; Spodoptera frugiperda JE Smith (Lepidoptera: Noctuidae)] is a generalist pest of many plant species and can feed on switchgrass as well. Here, in two different trials, FAW larval mass were significantly reduced when fed on lowland cultivar Kanlow relative to larvae fed on upland cultivar Summer plants after 10 days. Hormone content of plants indicated elevated levels of the plant defense hormone jasmonic acid (JA) and its bioactive conjugate JA-Ile although significant differences were not observed. Conversely, the precursor to JA, 12-oxo-phytodienoic acid (OPDA) levels were significantly different between FAW fed Summer and Kanlow plants raising the possibility of differential signaling by OPDA in the two cultivars. Global transcriptome analysis revealed a stronger response in Kanlow plant relative to Summer plants. Among these changes were a preferential upregulation of several branches of terpenoid and phenylpropanoid biosynthesis in Kanlow plants suggesting that enhanced biosynthesis or accumulation of antifeedants could have negatively impacted FAW larval mass gain on Kanlow plants relative to Summer plants. A comparison of the switchgrass-FAW RNA-Seq dataset to those from maize-FAW and switchgrass-aphid interactions revealed that key components of plant responses to herbivory, including induction of JA biosynthesis, key transcription factors and JA-inducible genes were apparently conserved in switchgrass and maize. In addition, these data affirm earlier studies with FAW and aphids that the cultivar Kanlow can provide useful genetics for the breeding of switchgrass germplasm with improved insect resistance.


Fc-receptor-mediated phagocytosis occurs in macrophages without an increase in average [Ca++]i.
The calcium ion has been implicated as a cytosolic signal or regulator in phagocytosis. Using the Ca++-sensitive photoprotein aequorin we have measured intracellular free Ca++ ion concentration ([Ca++]i) in thioglycolate-elicited mouse peritoneal macrophages during phagocytosis and IgG-induced spreading. Macrophages plated on glass were loaded with aequorin and [Ca++]i was then measured from cell populations, both as previously described (McNeil, P. L., and D. L. Taylor, 1985, Cell Calcium, 6:83-92). Aequorin indicated a resting [Ca++]i in adherent macrophages of 84 nM and was responsive to changes in [Ca++]i induced by the addition of Mg-ATP (0.1 mM) or serum to medium. However, during the 15 min required for phagocytosis of seven or eight IgG-coated erythrocytes per macrophage loaded with aequorin, we measured no change in [Ca++]i. Similarly, the ligation of Fc-receptors that occurs when macrophages spread on immune complex-coated coverslips did not change macrophage [Ca++]i. In contrast, a rise in [Ca++]i of macrophages was measured during phagocytosis occurring in a serum-free saline of pH 7.85, and as a consequence of incubation with quin2 A/M. We estimate that had a change in [Ca++]i occurred during phagocytosis, aequorin would have detected a rise from 0.1 to 1.0 microM taking place in as little as 2% of the macrophage's cytoplasmic volume. We therefore suggest that either Ca++ is not involved as a cytoplasmic signal for phagocytosis or that increases in [Ca++]i during phagocytosis are confined to such small regions of cytoplasm as to be below the limits of detection by our cellular averaging method. Our data emphasizes, moreover, the need for well-defined, nonperturbing conditions in such measurements of [Ca++]i.


A stochastic simulation model of U.S. breast cancer mortality trends from 1975 to 2000.
We present a simulation model that predicts U.S. breast cancer mortality trends from 1975 to 2000 and quantifies the impact of screening mammography and adjuvant therapy on these trends. This model was developed within the Cancer Intervention and Surveillance Network (CISNET) consortium. A Monte Carlo simulation is developed to generate the life history of individual breast cancer patients by using CISNET base case inputs that describe the secular trend in breast cancer risk, dissemination patterns for screening mammography and adjuvant treatment, and death from causes other than breast cancer. The model generates the patient's age, tumor size and stage at detection, mode of detection, age at death, and cause of death (breast cancer versus other) based in part on assumptions on the natural history of breast cancer. Outcomes from multiple birth cohorts are summarized in terms of breast cancer mortality rates by calendar year. Predicted breast cancer mortality rates follow the general shape of U.S. breast cancer mortality rates from 1975 to 1995 but level off after 1995 as opposed to following an observed decline. Sensitivity analysis revealed that the impact adjuvant treatment may be underestimated given the lack of data on temporal variation in treatment efficacy. We developed a simulation model that uses CISNET base case inputs and closely, but not exactly, reproduces U.S. breast cancer mortality rates. Screening mammography and adjuvant therapy are shown to have both contributed to a decline in U.S. breast cancer mortality.


Role of interleukin 6 (IL-6)/IL-6R-induced signal tranducers and activators of transcription and mitogen-activated protein kinase/extracellular.
Studies have described elevated levels of interleukin 6 (IL-6) and its soluble receptor (sIL-6R) in osteoarthritic and rheumatoid joints, as well as the inhibitory effect of this combination on cartilage matrix production. We investigated the ability of IL-6/sIL-6R to modulate gene expression of matrix metalloproteinase (MMP) and ADAMTS (ADAMS with thrombospondin motifs) family members in bovine chondrocytes, and the potential role of signal transducers and activators of transcription (STAT) and mitogen-activated protein kinases (MAPK) in this regulation. Primary cultures of bovine chondrocytes were stimulated with IL-6/sIL-6R for 30 min (Western blot and EMSA) or 24 h (RNA measurements) in the presence or absence of the STAT inhibitor parthenolide or the MAPK inhibitor PD 098059. mRNA was assessed by RT-PCR for the expression of MMP (MMP-1, -3, and -13) and 2 ADAMT family members (ADAMTS-4 and -5/11). IL-6/sIL-6R markedly induced activation of STAT and extracellular signal-related kinase (ERK1/2) and the subsequent expression of the collagenases MMP-1 and MMP-13 as well as MMP-3, an aggrecan-degrading enzyme and activator of pro-MMP. Expression of the 2 specific aggrecanases ADAMTS-4 and -5/11 was also elevated by this combination. Both STAT and MAPK signaling pathways were found to contribute to the IL-6/sIL-6R induction mechanisms, the overall effect being dependent on the respective magnitude of response and the crosstalk between the 2 pathways. These data indicate that the cartilage-degrading properties of IL-6/sIL-6R are mediated by induction of the aggrecan-degrading enzymes ADAMTS-4, -5/11, and MMP-3, and the collagen-degrading enzymes MMP-1 and -13. STAT and MAPK pathways play a crucial role in IL-6/sIL-6R modulation of these enzymes, suggesting that new strategies in the treatment of osteoarticular diseases might target these transduction cascades.


Estimating species distribution and abundance in river networks using environmental DNA.
All organisms leave traces of DNA in their environment. This environmental DNA (eDNA) is often used to track occurrence patterns of target species. Applications are especially promising in rivers, where eDNA can integrate information about populations upstream. The dispersion of eDNA in rivers is modulated by complex processes of transport and decay through the dendritic river network, and we currently lack a method to extract quantitative information about the location and density of populations contributing to the eDNA signal. Here, we present a general framework to reconstruct the upstream distribution and abundance of a target species across a river network, based on observed eDNA concentrations and hydro-geomorphological features of the network. The model captures well the catchment-wide spatial biomass distribution of two target species: a sessile invertebrate (the bryozoan Fredericella sultana) and its parasite (the myxozoan Tetracapsuloides bryosalmonae). Our method is designed to easily integrate general biological and hydrological data and to enable spatially explicit estimates of the distribution of sessile and mobile species in fluvial ecosystems based on eDNA sampling.


Industry ties and evidence in public comments on the FDA framework for modifications to artificial intelligence/machine learning-based medical devices: a cross sectional study.
To determine the extent and disclosure of financial ties to industry and use of scientific evidence in comments on a US Food and Drug Administration (FDA) regulatory framework for modifications to artificial intelligence/machine learning (AI/ML)-based software as a medical device (SaMD). Cross-sectional study. We searched all publicly available comments on the FDA 'Proposed Regulatory Framework for Modifications to Artificial Intelligence/Machine Learning (AI/ML)-Based Software as a Medical Device (SaMD)-Discussion Paper and Request for Feedback' from 2 April 2019 to 8 August 2019. The proportion of articles submitted by parties with financial ties to industry, disclosing those ties, citing scientific articles, citing systematic reviews and meta-analyses, and using a systematic process to identify relevant literature. We analysed 125 comments submitted on the proposed framework. 79 (63%) comments came from parties with financial ties; for 36 (29%) comments, it was not clear and the absence of financial ties could only be confirmed for 10 (8%) comments. No financial ties were disclosed in any of the comments that were not from industry submitters. The vast majority of submitted comments (86%) did not cite any scientific literature, just 4% cited a systematic review or meta-analysis and no comments indicated that a systematic process was used to identify relevant literature. Financial ties to industry were common and undisclosed, and scientific evidence, including systematic reviews and meta-analyses, were rarely cited. To ensure regulatory frameworks best serve patient interests, the FDA should mandate disclosure of potential conflicts of interest (including financial ties) in comments, encourage the use of scientific evidence, and encourage engagement from non-conflicted parties.


Green tea constituents (-)-epigallocatechin-3-gallate (EGCG) and gallic acid induce topoisomerase I- and topoisomerase II-DNA complexes in cells mediated by pyrogallol-induced hydrogen peroxide.
Green tea and its major active constituent, (-)-epigallocatechin-3-gallate (EGCG), are in clinical trials for the prevention and treatment of several diseases such as cancer. DNA topoisomerase (topo) poisons are commonly prescribed anticancer drugs that kill cancer cells by inducing topo-DNA complexes. Using purified topoisomerases, previous in vitro studies have shown that EGCG induces the formation of topo-DNA complexes. Because the activity of a drug on purified topoisomerases does not always represent the activity in a cell, we have used an immunofluorescence technique that allows the visualisation of topo I- and topo II-DNA complexes produced in individual cells to evaluate the activity of EGCG on both enzymes. High levels of topo I- and topo II-DNA complexes were observed in K562 leukaemia cells exposed to EGCG. Similar levels of topo I- and topo II-DNA complexes were visualised in cells treated with gallic acid (GA) (the acid part of the EGCG ester). Pyrogallol (PG) also induced topo-DNA complexes with both enzymes, therefore suggesting that the activity of EGCG and GA is mediated by their PG moieties. Catalase prevented both the cytotoxicity and the formation of topo I- and topo II-DNA complexes induced by EGCG, GA, PG and myricetin (a PG-containing flavonoid recently shown to induce topo I- and topo II-DNA complexes in cells), indicating that hydrogen peroxide mediates these activities. Hydrogen peroxide induced topo I- and topo II (α and β)-DNA complexes in a time- and dose-dependent manner. The formation of topo I- and topo II-DNA complexes in cells exposed to hydrogen peroxide correlated well with the induction of apoptosis, suggesting that the topo-DNA complexes induced at long exposure times by the compounds tested in our study may be apoptotic topo-DNA complexes. Finally, we report results suggesting that PG-containing drugs may selectively kill tumour cells by generating hydrogen peroxide.


EFFECTS OF A DIETARY SUPPLEMENT ON THE INCIDENCE OF ACUTE RESPIRATORY INFECTIONS IN SUSCEPTIBLE ADULTS: A RANDOMIZED CONTROLLED TRIAL.
although supplementation of specific micronutrients may improve immunologic factors, few studies about the combination of micronutrients with plant extracts on the occurrence of acute respiratory infections (ARI) have been published. to assess the effect of a nutritional supplement with micronutrients and plant extracts on the incidence of ARI in susceptible adults between January and April, 2012. a randomized, double-blind, parallel, placebo- controlled clinical trial was performed. Participants were adults susceptible to ARI who were healthy at the time of evaluation, signed informed consent forms and were not taking medication. They completed a medical history; weight, height, vital signs and laboratory analyses were assessed. Subjects were randomly assigned for consumption of the supplement or a placebo, for a 90 days period. Subjects made daily diary entries indicating the presence ARI symptoms. Those who became ill notified researchers and the attending physician confirmed the presence of an infection. Fisher's exact test was used to compare the proportion of ill subjects between groups. Relative risk and risk difference were also calculated (p < 0.05 significant). of 59 included subjects, 45 (25 women) completed the study (21 in the supplemented group and 24 in the placebo group). There were no significant differences at baseline between groups. After the intervention, the supplemented group had a lower incidence of ARI compared with the placebo group (57.1% vs. 91.7%, p = 0.013, RR = 0.62, 95%CI 0.42, 0.92). the consumption of a supplement with vitamins, minerals and plant extracts may decrease the incidence of ARI in susceptible adults.


Big data science: A literature review of nursing research exemplars.
Big data and cutting-edge analytic methods in nursing research challenge nurse scientists to extend the data sources and analytic methods used for discovering and translating knowledge. The purpose of this study was to identify, analyze, and synthesize exemplars of big data nursing research applied to practice and disseminated in key nursing informatics, general biomedical informatics, and nursing research journals. A literature review of studies published between 2009 and 2015. There were 650 journal articles identified in 17 key nursing informatics, general biomedical informatics, and nursing research journals in the Web of Science database. After screening for inclusion and exclusion criteria, 17 studies published in 18 articles were identified as big data nursing research applied to practice. Nurses clearly are beginning to conduct big data research applied to practice. These studies represent multiple data sources and settings. Although numerous analytic methods were used, the fundamental issue remains to define the types of analyses consistent with big data analytic methods. There are needs to increase the visibility of big data and data science research conducted by nurse scientists, further examine the use of state of the science in data analytics, and continue to expand the availability and use of a variety of scientific, governmental, and industry data resources. A major implication of this literature review is whether nursing faculty and preparation of future scientists (PhD programs) are prepared for big data and data science.


World Health Organization surveys to monitor HIV drug resistance prevention and associated factors in sentinel antiretroviral treatment sites.
The World Health Organization (WHO) estimates that >2 million people will have started antiretroviral therapy (ART) by the end of 2006. As the development of some HIV drug resistance (HIVDR) is inevitable in populations taking ART, the emergence of HIVDR must be balanced against the benefits of providing ART, including improved health outcomes and decreased HIV/AIDS-associated morbidity and mortality. ART programmes should operate to minimize the emergence of HIVDR in populations receiving therapy and HIVDR itself must be monitored to ensure ongoing regimen efficacy. ART regimens in resource-limited settings are usually selected at the national level following a public health approach: generally only one first-line regimen with alternate regimen(s) incorporating within-class drug substitutions are available in the public sector. The WHO has developed a population-based HIVDR assessment and prevention strategy, which includes standardized HIVDR monitoring surveys in populations receiving first-line ART at sentinel sites. The WHO surveys monitor HIVDR prevention in sentinel sites by utilizing a standardized, minimum-resource prospective survey methodology to assess the success of adult and paediatric ART sites in preventing HIVDR emergence during the first year of ART. The surveys also identify associated factors that can be addressed at the level of the ART site or programme. WHO HIVDR monitoring surveys are designed to be integrated easily into a country's ongoing, routine HIV-related evaluation activities. Performed regularly at representative sites, the data generated will inform evidence-based decision making regarding national and global ART regimen selection and minimize the emergence of HIVDR at a population level.


Generalized scattering-matrix method for the analysis of two-dimensional photonic bandgap devices.
Development of accurate numerical methods for the analysis of photonic-bandgap-based devices is a relevant issue in optimizing existing devices and/or developing new design solutions. Within this framework, we present an innovative and general approach for the evaluation of the electromagnetic behavior of two-dimensional finite-extent photonic crystals made of a finite set of parallel rods. The proposed approach is a generalization of the scattering-matrix method introduced by Maystre and co-workers and of its improved version proposed by the present authors, which exploits a suitable aggregation into "macrocells" to achieve a reduction of the number of unknowns. As a matter of fact, both of these approaches can be exploited only in those cases in which particular modal expansions for the fields hold true. In order to overcome this limitation, we propose a suitable exploitation of the method of auxiliary sources to provide a general and reliable method for the numerical computation of the scattering matrix of an object of arbitrary shape. By taking advantage of this, we can then generalize our improved scattering matrix method to further increase its computational effectiveness. A numerical analysis of some square-lattice configurations is reported to confirm the accuracy of the proposed method and the remarkable computational benefit.


Infinite kinetic stability against dissociation of supramolecular protein complexes through donor strand complementation.
Adhesive type 1 pili from uropathogenic Escherichia coli strains are heat and denaturant resistant, filamentous protein complexes. Individual pilus subunits associate through "donor strand complementation," whereby the incomplete immunoglobulin-like fold of each subunit is completed by the N-terminal extension of a neighboring subunit. We show that antiparallel donor strand insertion generally causes nonequilibrium behavior in protein folding and extreme activation energy barriers for dissociation of subunit-subunit complexes. We identify the most kinetically stable, noncovalent protein complex known to date. The complex between the pilus subunit FimG and the donor strand peptide of the subunit FimF shows an extrapolated dissociation half-life of 3 x 10(9) years. The 15 residue peptide forms ideal intermolecular beta sheet H-bonds with FimG over 10 residues, and its hydrophobic side chains strongly interact with the hydrophobic core of FimG. The results show that kinetic stability and nonequilibrium behavior in protein folding confers infinite stability against dissociation in extracellular protein complexes.


[Antiretroviral therapy].
Antiretroviral therapy to treat HIV, as we know it today, is nothing less than a huge success story in modern medical history. What used to be an almost certain death-sentence was transformed into a very manageable chronic disease by means of highly efficient und mostly well tolerated drugs. Today, HIV-infected patients treated according to international recommendations have a very good chance to outgo the negative effects of HIV-1 and are therefore able to reach an almost normal life expectancy. Furthermore, patients successfully treated with antiretroviral drugs are no longer infectious, which is an essential aspect of global strategies to overcome the pandemic. Nevertheless, due to the complexity of HIV, physicians treating patients with antiretroviral therapy require profound knowledge of aspects such as viral resistance mechanisms and immune reconstitution, as well as drug-toxicity und drug-drug-interactions. Many other aspects such as long-term side-effects of antiretroviral drugs are still unknown. Strict adherence to treatment is of utmost importance.


Religious circumcision (Khatna) and circumcision mishaps in Kashmiri children.
Male circumcision is recommended practice in Muslim tradition and one of the oldest operations performed all over the world. Male circumcision is universal in our Muslim-dominated valley of Kashmir for religious reasons. It can be performed by different techniques such as the conventional open methods, the device methods and sutureless methods. The objective of this study was to report our practice of male circumcision amongst children and compare the different common surgical techniques and highlight the circumcision mishaps conducted by quacks in the Kashmir Valley. This was a comparative observational study conducted at SKIMS Medical College and Hospital, from 2017 to 2021. Children who presented for primary circumcision were subjected to one of two different surgical techniques; the dorsal slit or Guillotine method. The prospective analysis of children managed for circumcision mishaps conducted by non-professionals was also included in the study. Circumcisions done after 2 years of age were defined as delayed. The data were collected analysed using SPSS software (SPSS version 22, IBM, Armonk, NY, USA). Total of 689 Kashmiri Muslim male children between 1 day and 10 years of age over a period of 5 years were studied. Six hundred and fifty-five children for primary circumcision and 34 children managed for circumcision mishaps were included in the study. Amongst the subjects for primary circumcision, the most number of children were between 1 and 2 years of age (33.28%). One hundred and fourteen (25.73%) hospital-delivered babies and 201 (94.81%) home-delivered babies had delayed circumcision that is after 2 years of age (P = 0.00001). Religious requirement was the only indication for circumcision in this study. Three hundred and ninety-six (60.46%) children were circumcised with dorsal slit and 259 (39.54%) with guillotine method using computer-generated random numbers. Complications were found 8.08% of subjects in dorsal slit method as compared to 16.60% in guillotine technique (P = 0.008). Out of 34 children managed for circumcision mishaps, 11 (32.35%) presented with massive bleeding after primary circumcision by half doctors, 18 (52.94%) had incomplete circumcision, 3 (8.82%) had multiple skin bridges and 2 (5.88%) had incomplete circumcision with glans injury. All the patients with circumcision mishaps were treated with good outcome. There was no mortality. Circumcision occurs at a wide range of ages and male circumcision is universal in our Muslim-dominated valley for religious reasons. Circumcision by quacks and the associated complications are still prevalent in our society. The procedure is safe and free of any major complications when conducted by trained medical personnel under aseptic conditions of the operation theatre and hence should be encouraged.


The effectiveness of dopamine agonists for treatment of neuropsychiatric symptoms post brain injury and stroke.
Traumatic brain injury and stroke are among the leading causes of neurological disability worldwide. Although dopaminergic agents have long been associated with improvement of neuropsychiatric outcomes, to date much of the evidence to date has been in case reports and case series or open label trials. We undertook a systematic review of double-blinded randomised controlled trials (RCT) to determine the effect of dopaminergic agents on pre-defined outcomes of (a) apathy; (b) psychomotor retardation; (c) behavioural management and (d) cognitive function. Databases searched were: Medline, EMBASE, and PsychInfo for human studies. The Cochrane Clinical Trials Database and the TRIP Medical database were also searched. All identified studies, were further hand-searched. We identified six studies providing data on 227 participants, 150 of whom received dopaminergic therapy. Trials were compromised by cross-over design, inadequate wash out period, small numbers and heterogeneous outcome measures. However one good quality RCT demonstrates the efficacy of amantadine in behavioural management. One further RCT shows methylphenidate-levodopa is efficacious for mood post-stroke. One study shows rotigotine to improve hemi-inattention caused by prefrontal damage. Our systematic review demonstrates an evolving evidence base to suggest some benefits in agitation and aggression, mood and attentional deficits. However, there are key limitations of the studies undertaken to date involving small numbers of participants, heterogeneous outcome measures, and variable study designs. There is a need for on-going large prospective double-blind RCTs in these medications using standardised criteria and outcomes to fully understand their effectiveness in this patient group.


Training needs of healthcare providers related to Centers for Disease Control and Prevention core competencies for fetal alcohol spectrum disorders.
Fetal alcohol spectrum disorders (FASDs) are birth defects directly linked to consumption of alcohol during pregnancy and hence completely preventable. Many health and allied health professionals are in prime positions for primary prevention of FASDs through work with women of childbearing age and secondary prevention through work with affected individuals whose lives can be greatly improved via tailored intervention. To develop educational guidelines for FASD prevention. Interviews were conducted with 26 individuals representing eight health or allied health professions. Participants were asked about professional groups with which they had sufficient experience to describe FASD-related competencies and educational needs for the given group(s). For each group, participants were asked for their perceptions of group members' FASD awareness, knowledge, and skills application as related to the seven core competencies for FASD practice developed by the Centers for Disease Control and Prevention (CDC). Findings revealed that competence, especially when viewed separately in terms of knowledge versus capacity for application of information, in the area of FASDs is unevenly distributed among and throughout healthcare provider groups. Based on this information, recommendations are offered for optimal health and allied health education efforts to prevent and treat FASDs, framed along FASD core competencies recommended by the CDC.


The phosphorylation specificity of B-RAF WT, B-RAF D594V, B-RAF V600E and B-RAF K601E kinases: an in silico study.
Phosphorylation of the B-RAF kinase is an essential process in tumour induction and maintenance in several cancers. Herein the phosphorylation specificity of the activation segment of the wild type B-RAF kinase and the B-RAF(D594V), B-RAF(V600E) and B-RAF(K601E) mutants was examined by molecular dynamics (MD) simulations and GRID molecular interaction field analysis. According to our analysis, Thr599 and Ser602 were the only residues in the activation segment in B-RAF(WT) that were well exposed to ATP binding, which is in agreement with the experimental results, and provide a molecular basis of the observed phosphorylation. The phosphorylation specificity was altered significantly for each of the three different mutants studied due to the large conformational changes and subsequent alterations in the electrostatic forces between several residues for each of these mutants. Thus the analysis revealed limited phosphorylation potential of the non-active B-RAF(D594V) mutant and several potential ATP binding sites were identified for the highly active B-RAF(V600E) mutant. The Lys601 residue, which is specific to RAF and not present in the activation segment of other similar kinases, was identified to potentially be of major importance to the observed differences in the phosphorylation specificity of the mutants. Our results indicate that Lys601 might be a specific ATP coordinating residue, contributing to the B-RAF phosphorylation specificity. The overall results can be helpful for the understanding of the B-RAF phosphorylation processes on a molecular level.


Toward Human-Carnivore Coexistence: Understanding Tolerance for Tigers in Bangladesh.
Fostering local community tolerance for endangered carnivores, such as tigers (Panthera tigris), is a core component of many conservation strategies. Identification of antecedents of tolerance will facilitate the development of effective tolerance-building conservation action and secure local community support for, and involvement in, conservation initiatives. We use a stated preference approach for measuring tolerance, based on the 'Wildlife Stakeholder Acceptance Capacity' concept, to explore villagers' tolerance levels for tigers in the Bangladesh Sundarbans, an area where, at the time of the research, human-tiger conflict was severe. We apply structural equation modeling to test an a priori defined theoretical model of tolerance and identify the experiential and psychological basis of tolerance in this community. Our results indicate that beliefs about tigers and about the perceived current tiger population trend are predictors of tolerance for tigers. Positive beliefs about tigers and a belief that the tiger population is not currently increasing are both associated with greater stated tolerance for the species. Contrary to commonly-held notions, negative experiences with tigers do not directly affect tolerance levels; instead, their effect is mediated by villagers' beliefs about tigers and risk perceptions concerning human-tiger conflict incidents. These findings highlight a need to explore and understand the socio-psychological factors that encourage tolerance towards endangered species. Our research also demonstrates the applicability of this approach to tolerance research to a wide range of socio-economic and cultural contexts and reveals its capacity to enhance carnivore conservation efforts worldwide.


21st Century Genetics: Mass Spectrometry of Yeast Telomerase.
Telomerase is a specialized reverse transcriptase that maintains the ends of chromosomes in almost all eukaryotes. The core of telomerase consists of telomerase RNA and the reverse transcriptase that uses a short segment without the RNA to template the addition of telomeric repeats. In addition, one or more accessory proteins are required for telomerase action in vivo. The best-studied accessory protein is Est1, which is conserved from yeasts to humans. In budding yeast, Est1 has two critical in vivo functions: By interaction with Cdc13, a telomere-binding protein, it recruits telomerase to telomeres, and it also increases telomerase activity. Although budding yeast telomerase is highly regulated by the cell cycle, Est1 is the only telomerase subunit whose abundance is cell cycle-regulated. Close to 400 yeast genes are reported to affect telomere length, although the specific function of most of them is unknown. With the goal of identifying novel telomerase regulators by mass spectrometry, we developed methods for purifying yeast telomerase and its associated proteins. We summarize the methods we used and describe the experiments that show that four telomerase-associated proteins identified by mass spectrometry, none of which had been linked previously to telomeres, affect telomere length and cell cycle regulation of telomerase by controlling Est1 abundance.


Reproductive health differences among Latin American- and US-born young women.
Investigations of reproductive health within Latinos living in the United States suggest that sexual behaviors and contraception use practices vary by ethnicity and between foreign- and US-born adolescents. This article compares high-risk sexual behaviors and reproductive health among foreign-born Latinas, US-born Latinas, and US-born non-Latinas aged 15-24 years. We recruited 361 females from reproductive health clinics in the San Francisco Bay Area of California between 1995 and 1998; these women completed an interview that assessed sexual risk behaviors and history of pregnancy, abortion, and sexually transmitted infections. Current chlamydial and gonococcal infections were detected through biological testing. Among participants aged 15-18 years, US-born Latinas were more likely to have been pregnant (odds ratio [OR] comparing US-born Latinas and US-born non-Latinas = 3.9, 95% confidence interval [CI] 1.3, 11.4), whereas among respondents aged 19-24 years, foreign-born Latinas were more likely to have been pregnant than US-born Latinas (OR = 11.3, 95% CI 1.0, 130.8) and US-born non-Latinas (OR = 64.2, 95% CI 9.9, 416.3). US-born Latinas were most likely to have had an abortion (OR comparing US-born Latinas and US-born non-Latinas = 2.0, 95% CI 0.9, 4.7). They were also most likely to have chlamydial infection at study enrollment (8.2% prevalence compared to 2.2% and 1.0%for foreign-born Latinas and US-born non-Latinas, respectively; P =.009). Reproductive health differences between foreign and US-born females and within the US-born population warrant further examination and highlight the need for targeted prevention.


Nutrient Patterns and Risk of Polycystic Ovary Syndrome.
There are limited data on the role of nutrient patterns in development of polycystic ovary syndrome (PCOS). The aim of the study is to document the relationship between nutrient patterns and PCOS. In this study, 281 incident PCOS women and 472 controls were interviewed through the endocrine clinics between February 2013 and March 2015 in Tehran, Iran. Usual dietary intakes were obtained using a validated semi-quantitative food frequency questionnaire. Factor analysis was conducted on the basis of 32 nutrients. Unconditional logistic regression was performed to ascertain odds ratios. The p<0.05 was considered for significance level. In principal component analysis two nutrient patterns emerged. Factor 1 had high loadings for riboflavin, niacin, pyridoxine, thiamin, magnesium, pantothenic acid, cobalamin, vitamin C, folate, vitamin D, total fiber, selenium, phosphorus, vitamin E, manganese, vitamin K, monounsaturated fatty acids, polyunsaturated fatty acids, potassium and vegetable protein. Factor 2 characterized by high loadings for carbohydrate, animal protein, fat, cholesterol, saturated fatty acid, sodium, biotin, copper, iron, fluoride, zinc, and calcium. After adjusting for potential confounders, the risk of PCOS was significantly higher in the highest tertile of factor 2 (OR: 2.38, 95% CI: 1.69-3.21). Conversely, being in the highest tertile of factor 1 was associated with a lower risk of PCOS (OR: 0.48, 95% CI: 0.21-0.82). Our results provide a possible new insight into the interactions between nutrient intakes and PCOS.


The trends of disease burden due to high temperature in Mainland China from 1990 to 2019 and its prediction to 2030.
The risk of high-temperature-related diseases is increasing owing to global warming. This study aimed to assess the trend of disease burden caused by high temperatures in Mainland China from 1990 to 2019 and to predict the trend of disease burden over the next 10 years. The latest data were downloaded from the Global Burden of Disease Database (GBD) for analysis, and the disease burden related to high temperature was described by mortality and disability-adjusted life-years (DALYs) and stratified by etiology, sex, and age. Statistical analyses were performed using the R software. In 2019, there were 13,907 deaths attributed to high temperatures in Mainland China, and this was 29.55% higher than the 10,735 deaths in 1990. Overall, the age-standardized mortality and DALYs attributed to high temperatures showed a downward trend from 1990 to 2019. We observed an etiological shift in high-temperature-related diseases. The age-standardized DALYs contribution attributed to high temperatures in 1990 was mainly from communicable, maternal, neonatal, and nutritional diseases (CMNND) (21.81/100,000), followed by injury (18.30/100,000) and non-communicable diseases (10.40/100,000). In 2019, the largest contribution shifted to non-communicable diseases (10.07/100,000), followed by injuries (5.21/100,000), and CMNND (2.30/100,000). The disease burden attributed to high temperatures was higher in males than in females and increased with age. In 2030, the mortality rate and DALYs due to high temperatures are predicted to decrease further, and the largest contribution will come from chronic non-communicable diseases, the occurrence of which will remain at a high level over the next 10 years. The burden of disease due to high temperatures in Mainland China is still heavy, mainly due to population aging and an increase in non-communicable diseases.


Finding common protein interaction patterns across organisms.
Protein interactions are an important resource to obtain an understanding of cell function. Recently, researchers have compared networks of interactions in order to understand network evolution. While current methods first infer homologs and then compare topologies, we here present a method which first searches for interesting topologies and then looks for homologs. PINA (protein interaction network analysis) takes the protein interaction networks of two organisms, scans both networks for subnetworks deemed interesting, and then tries to find orthologs among the interesting subnetworks. The application is very fast because orthology investigations are restricted to subnetworks like hubs and clusters that fulfill certain criteria regarding neighborhood and connectivity. Finally, the hubs or clusters found to be related can be visualized and analyzed according to protein annotation.


RINGO C is required to sustain the spindle-assembly checkpoint.
RINGO/Speedy proteins are direct activators of Cdk1 and Cdk2 that have no sequence homology to cyclins. We have characterized the role in cell-cycle progression of a new human member of this protein family referred to as RINGO C. We show that siRNA-mediated knockdown of RINGO C results in premature mitotic exit with misaligned chromosomes, even in the presence of microtubule poisons. Time-lapse-microscopy experiments suggest that RINGO C is involved in the spindle-assembly checkpoint (SAC). Consistent with this idea, RINGO-C-depleted cells show impaired recruitment of the SAC components Mad2, Bub1 and BubR1. As the checkpoint is overridden, cells display defective chromosome segregation, which leads to an increased number of micronuclei and binucleated structures. Intriguingly, we found that RINGO C can associate with the mitotic kinase Aurora B, and downregulation of RINGO C produces mislocalization of the active form of Aurora B in prometaphase. Taken together, our results indicate a role for RINGO C in the mitotic checkpoint, which might be mediated by defective recruitment of SAC components and deregulation of the activity of Aurora kinase B.


Single versus multiple-family intervention in childhood overweight--Finnmark Activity School: a randomised trial.
To compare a new comprehensive lifestyle programme performed in groups of families with overweight (included obese) children with a more conventional single-family programme. The study design and interim anthropometrical results after 12 months are presented. Altogether 97 overweight and obese children aged 6-12 years with body mass index (BMI) corresponding to cut-off point ≥ 27.5 in adults were included. Study participants were randomised to multiple-family intervention (MUFI) or single-family intervention (SIFI) in a parallel design. MUFI comprised a 3-day inpatient programme at the hospital with other families and a multidisciplinary team, follow-up visits in their hometown individually and in groups, organised physical activity twice weekly and a 4-day family camp after 6 months. SIFI comprised individual counselling by paediatric nurse, paediatric consultant and nutritionist at the hospital and follow-up by public health nurse in the community. Solution focused approach was applied in both interventions. Primary outcome measures were change in BMI kg/m(2) and BMI SD score (BMI SDS). BMI increased by 0.37 units in the MUFI compared to 0.77 units in the SIFI (p=0.18). BMI SDS decreased by 0.16 units in the MUFI group compared to 0.07 units in the SIFI group (p=0.07). Secondary endpoint waist circumference decreased 0.94 cm in the multiple-family group and increased 0.95 cm in the single-family group, p=0.04. Interim analysis after 12 months showed no between-group difference in terms of BMI or BMI SDS. The MUFI group had a significant decrease in waist circumference compared to the SIFI group. at http://www.clinicaltrials.gov (NCT00872807).


Cloning and characterization of a selenium-independent glutathione peroxidase (HC29) from adult Haemonchus contortus.
The complete coding sequence of Haemonchus (H.) contortus HC29 cDNA was generated by rapid amplification of cDNA ends in combination with PCR using primers targeting the 5'- and 3'-ends of the partial mRNA sequence. The cloned HC29 cDNA was shown to be 1,113 bp in size with an open reading frame of 507 bp, encoding a protein of 168 amino acid with a calculated molecular mass of 18.9 kDa. Amino acid sequence analysis revealed that the cloned HC29 cDNA contained the conserved catalytic triad and dimer interface of selenium-independent glutathione peroxidase (GPX). Alignment of the predicted amino acid sequences demonstrated that the protein shared 44.7~80.4% similarity with GPX homologues in the thioredoxin-like family. Phylogenetic analysis revealed close evolutionary proximity of the GPX sequence to the counterpart sequences. These results suggest that HC29 cDNA is a GPX, a member of the thioredoxin-like family. Alignment of the nucleic acid and amino acid sequences of HC29 with those of the reported selenium-independent GPX of H. contortus showed that HC29 contained different types of spliced leader sequences as well as dimer interface sites, although the active sites of both were identical. Enzymatic analysis of recombinant prokaryotic HC29 protein showed activity for the hydrolysis of H(2)O(2). These findings indicate that HC29 is a selenium-independent GPX of H. contortus.


Mechanical characterization and numerical simulation of a subcutaneous implantable 3D printed cell encapsulation system.
Cell transplantation in bioengineered scaffolds and encapsulation systems has shown great promise in regenerative medicine. Depending on the site of implantation, type of cells and their expected function, these systems are designed to provide cells with a physiological-like environment while providing mechanical support and promoting long-term viability and function of the graft. A minimally invasive 3D printed system termed neovascularized implantable cell homing and encapsulation (NICHE) was developed in polylactic acid for subcutaneous transplantation of endocrine cells, including pancreatic islets. The suitability of the NICHE for long term in vivo deployment is investigated by assessing mechanical behavior of both fresh devices under simulated subcutaneous conditions and NICHE retrieved from subcutaneous implantation in pigs. Both experimental and numerical studies were performed with a focus on validating the constitutive material model used in the numerical analysis for accuracy and reliability. Notably, homogeneous isotropic constitutive material model calibrated by means of uniaxial testing well suited experimental results. The results highlight the long term durability for in vivo applications and the potential applicability of the model to predict the mechanical behavior of similar devices in various physiological settings.


Representation of cognitive reappraisal goals in frontal gamma oscillations.
Recently, numerous efforts have been made to understand the neural mechanisms underlying cognitive regulation of emotion, such as cognitive reappraisal. Many studies have reported that cognitive control of emotion induces increases in neural activity of the control system, including the prefrontal cortex and the dorsal anterior cingulate cortex, and increases or decreases (depending upon the regulation goal) in neural activity of the appraisal system, including the amygdala and the insula. It has been hypothesized that information about regulation goals needs to be processed through interactions between the control and appraisal systems in order to support cognitive reappraisal. However, how this information is represented in the dynamics of cortical activity remains largely unknown. To address this, we investigated temporal changes in gamma band activity (35-55 Hz) in human electroencephalograms during a cognitive reappraisal task that was comprised of three reappraisal goals: to decease, maintain, or increase emotional responses modulated by affect-laden pictures. We examined how the characteristics of gamma oscillations, such as spectral power and large-scale phase synchronization, represented cognitive reappraisal goals. We found that left frontal gamma power decreased, was sustained, or increased when the participants suppressed, maintained, or amplified their emotions, respectively. This change in left frontal gamma power appeared during an interval of 1926 to 2453 ms after stimulus onset. We also found that the number of phase-synchronized pairs of gamma oscillations over the entire brain increased when participants regulated their emotions compared to when they maintained their emotions. These results suggest that left frontal gamma power may reflect cortical representation of emotional states modulated by cognitive reappraisal goals and gamma phase synchronization across whole brain regions may reflect emotional regulatory efforts to achieve these goals. Our study may provide the basis for an electroencephalogram-based neurofeedback system for the cognitive regulation of emotion.


Cutaneous factors related to the risk of malignant melanoma.
In a case-control study, 287 women with malignant melanoma were compared with 574 age-matched controls. Red hair colour at age 5 years was associated with a tripling of risk [relative risk (RR) = 3.0], blonde hair with a 60% increase (RR = 1.6) and fair skin with a doubling (RR = 2.1). Women with melanoma also reported that they tended to burn (RR = 1.4) and to freckle (RR = 1.9) after exposure to sunlight. Since fair skin, red hair, and the tendency to burn or freckle after exposure to sunlight all cluster in the same individuals, the extent to which each of these factors had an independent influence on susceptibility to melanoma was investigated. Hair colour, especially red hair, proved to be the major determinant, followed by skin colour. The reporting of above average numbers of naevi on the body was as strong a predictor of melanoma as was red hair colour (RR = 3.4). A history of psoriasis was also more common in cases than controls (RR = 3.0) as was a history of vitiligo (RR = 1.8). A history of acne appeared to be protective (RR = 0.4) as did a history suggestive of chloasma (RR = 0.6) and premature greying of the hair (RR = 0.6). These relationships were irrespective of hair and skin colour.


The innate mononuclear phagocyte network depletes B lymphocytes through Fc receptor-dependent mechanisms during anti-CD20 antibody immunotherapy.
Anti-CD20 antibody immunotherapy effectively treats non-Hodgkin's lymphoma and autoimmune disease. However, the cellular and molecular pathways for B cell depletion remain undefined because human mechanistic studies are limited. Proposed mechanisms include antibody-, effector cell-, and complement-dependent cytotoxicity, the disruption of CD20 signaling pathways, and the induction of apoptosis. To identify the mechanisms for B cell depletion in vivo, a new mouse model for anti-CD20 immunotherapy was developed using a panel of twelve mouse anti-mouse CD20 monoclonal antibodies representing all four immunoglobulin G isotypes. Anti-CD20 antibodies rapidly depleted the vast majority of circulating and tissue B cells in an isotype-restricted manner that was completely dependent on effector cell Fc receptor expression. B cell depletion used both FcgammaRI- and FcgammaRIII-dependent pathways, whereas B cells were not eliminated in FcR common gamma chain-deficient mice. Monocytes were the dominant effector cells for B cell depletion, with no demonstrable role for T or natural killer cells. Although most anti-CD20 antibodies activated complement in vitro, B cell depletion was completely effective in mice with genetic deficiencies in C3, C4, or C1q complement components. That the innate monocyte network depletes B cells through FcgammaR-dependent pathways during anti-CD20 immunotherapy has important clinical implications for anti-CD20 and other antibody-based therapies.


Consultation-based academic interventions for children with ADHD: effects on reading and mathematics achievement.
The purpose of this investigation was to evaluate the relative efficacy of two consultation-based models for designing academic interventions to enhance the educational functioning of children with attention-deficit/hyperactivity disorder (ADHD). Children (N=167) meeting DSM-IV criteria for ADHD were randomly assigned to one of two consultation groups: Individualized Academic Intervention (IAI; interventions designed using a data-based decision-making model that involved ongoing feedback to teachers) and Generic Academic Intervention (GAI; interventions designed based on consultant-teacher collaboration, representing "consultation as usual"). Teachers implemented academic interventions over 15 months. Academic outcomes (e.g., standardized achievement test, and teacher ratings of academic skills) were assessed on four occasions (baseline, 3 months, 12 months, 15 months). Hierarchical linear modeling analyses indicated significant positive growth for 8 of the 14 dependent variables; however, trajectories did not differ significantly across consultation groups. Interventions in the IAI group were delivered with significantly greater integrity; however, groups did not differ with respect to teacher ratings of treatment acceptability. The results of this study provide partial support for the effectiveness of consultation-based academic interventions in enhancing educational functioning in children with ADHD; however, the relative advantages of an individualized model over "consultation as usual" have yet to be established.


Plesiomonas enteric infections in the United States.
Thirty-one persons nationwide from whom Plesiomonas shigelloides was isolated in large numbers from stool in 1984 were compared with 62 matched control subjects. Infection with P. shigelloides was strongly associated with eating uncooked shellfish, usually raw oysters, in the 48 hours before the onset of illness (p less than 0.00001) and with foreign travel (p less than 0.00006), usually to Mexico. Most ill persons had self-limited diarrhea with blood and mucus in stool and other clinical findings that suggested enteroinvasiveness of infecting organisms. Two patients developed their illnesses after taking ampicillin for reasons unrelated to diarrhea; plesiomonads recovered from their stools were resistant to ampicillin. Seven persons with gastrointestinal complaints had alleviation or resolution of their symptoms after taking antimicrobial agents to which recovered plesiomonads were susceptible. These findings suggest that P. shigelloides may cause enteric disease in the normal host, that it may be acquired from eating uncooked shellfish, and that it may be a cause of travelers' diarrhea.


View-independent action recognition from temporal self-similarities.
This paper addresses recognition of human actions under view changes. We explore self-similarities of action sequences over time and observe the striking stability of such measures across views. Building upon this key observation, we develop an action descriptor that captures the structure of temporal similarities and dissimilarities within an action sequence. Despite this temporal self-similarity descriptor not being strictly view-invariant, we provide intuition and experimental validation demonstrating its high stability under view changes. Self-similarity descriptors are also shown to be stable under performance variations within a class of actions when individual speed fluctuations are ignored. If required, such fluctuations between two different instances of the same action class can be explicitly recovered with dynamic time warping, as will be demonstrated, to achieve cross-view action synchronization. More central to the current work, temporal ordering of local self-similarity descriptors can simply be ignored within a bag-of-features type of approach. Sufficient action discrimination is still retained in this way to build a view-independent action recognition system. Interestingly, self-similarities computed from different image features possess similar properties and can be used in a complementary fashion. Our method is simple and requires neither structure recovery nor multiview correspondence estimation. Instead, it relies on weak geometric properties and combines them with machine learning for efficient cross-view action recognition. The method is validated on three public data sets. It has similar or superior performance compared to related methods and it performs well even in extreme conditions, such as when recognizing actions from top views while using side views only for training.


Ethanolic Extract of Orthosiphon stamineus Improves Memory in Scopolamine-Induced Amnesia Model.
Alzheimer's disease (AD) is a chronic neurodegenerative brain disease which is characterized by impairment in cognitive functioning. Orthosiphon stamineus (OS) Benth. (Lamiaceae) is a medicinal plant found around Southeast Asia that has been employed as treatments for various diseases. OS extract contains many active compounds that have been shown to possess various pharmacological properties whereby in vitro studies have demonstrated neuroprotective as well as cholinesterase inhibitory effects. This study, therefore aimed at determining whether this Malaysian plant derived flavonoid can reverse scopolamine induced learning and memory dysfunction in the novel object recognition (NOR) test and the elevated plus maze (EPM) test. In the present study, rats were treated once daily with OS 50 mg/kg, 100 mg/kg, 200 mg/kg and donepezil 1 mg/kg via oral dosing and were given intraperitoneal (ip) injection of scopolamine 1 mg/kg daily to induce cognitive deficits. Rats were subjected to behavioral analysis to assess learning and memory functions and hippocampal tissues were extracted for gene expression and immunohistochemistry studies. All the three doses demonstrated improved scopolamine-induced impairment by showing shortened transfer latency as well as the higher inflexion ratio when compared to the negative control group. OS extract also exhibited memory-enhancing activity against chronic scopolamine-induced memory deficits in the long-term memory novel object recognition performance as indicated by an increase in the recognition index. OS extract was observed to have modulated the mRNA expression of CREB1, BDNF, and TRKB genes and pretreatment with OS extract were observed to have increased the immature neurons against hippocampal neurogenesis suppressed by scopolamine, which was confirmed by the DCX-positive stained cells. These research findings suggest that the OS ethanolic extract demonstrated an improving effect on memory and hence could serve as a potential therapeutic target for the treatment of neurodegenerative diseases like AD.


Identification of a novel mono-leucine basolateral sorting motif within the cytoplasmic domain of amphiregulin.
Epithelial cells establish apical and basolateral (BL) membranes with distinct protein and lipid compositions. To achieve this spatial asymmetry, the cell utilizes a variety of mechanisms for differential sorting, delivery and retention of cell surface proteins. The EGF receptor (EGFR) and its ligand, amphiregulin (AREG), are transmembrane proteins delivered to the BL membrane in polarized epithelial cells. Herein, we show that the cytoplasmic domain of AREG (ACD) contains dominant BL sorting information; replacement of the cytoplasmic domain of apically targeted nerve growth factor receptor with the ACD redirects the chimera to the BL surface. Using sequential truncations and site-directed mutagenesis of the ACD, we identify a novel BL sorting motif consisting of a single leucine C-terminal to an acidic cluster (EEXXXL). In adaptor protein (AP)-1B-deficient cells, newly synthesized AREG is initially delivered to the BL surface as in AP-1B-expressing cells. However, in these AP-1B-deficient cells, recycling of AREG back to the BL surface is compromised, leading to its appearance at the apical surface. These results show that recycling, but not delivery, of AREG to the BL surface is AP-1B dependent.


A framework for evolutionary systems biology.
Many difficult problems in evolutionary genomics are related to mutations that have weak effects on fitness, as the consequences of mutations with large effects are often simple to predict. Current systems biology has accumulated much data on mutations with large effects and can predict the properties of knockout mutants in some systems. However experimental methods are too insensitive to observe small effects. Here I propose a novel framework that brings together evolutionary theory and current systems biology approaches in order to quantify small effects of mutations and their epistatic interactions in silico. Central to this approach is the definition of fitness correlates that can be computed in some current systems biology models employing the rigorous algorithms that are at the core of much work in computational systems biology. The framework exploits synergies between the realism of such models and the need to understand real systems in evolutionary theory. This framework can address many longstanding topics in evolutionary biology by defining various 'levels' of the adaptive landscape. Addressed topics include the distribution of mutational effects on fitness, as well as the nature of advantageous mutations, epistasis and robustness. Combining corresponding parameter estimates with population genetics models raises the possibility of testing evolutionary hypotheses at a new level of realism. EvoSysBio is expected to lead to a more detailed understanding of the fundamental principles of life by combining knowledge about well-known biological systems from several disciplines. This will benefit both evolutionary theory and current systems biology. Understanding robustness by analysing distributions of mutational effects and epistasis is pivotal for drug design, cancer research, responsible genetic engineering in synthetic biology and many other practical applications.


Antihypertensive effect of rice protein hydrolysate with in vitro angiotensin I-converting enzyme inhibitory activity in spontaneously hypertensive rats.
Angiotensin I-converting enzyme (ACE) plays a crucial role in the regulation of blood pressure as well as cardiovascular function. ACE catalyzes the conversion of angiotensin I to vasoconstrictor angiotensin II, and also inactivates the antihypertensive vasodilator bradykinin. Inhibition of ACE mainly results an overall antihypertensive effect. Food protein-derived peptides can have ACE-inhibiting properties and thus may be used as a novel functional food for preventing hypertension as well as for therapeutic purposes. In the present study, rice protein was hydrolyzed by protease Alcalase for 2 h and the resulted hydrolysate was determined for ACE inhibitory activity in vitro. The antihypertensive effect of rice protein hydrolysate was also investigated in spontaneously hypertensive rats (SHR). The Alcalase-generated hydrolysate showed strong in vitro ACE inhibitory activity with the IC50 value of 0.14 mg/ml. A significant decrease in systolic blood pressure in spontaneously hypertensive rats was observed following single oral administration of this hydrolysate at a dose of 600 mg/kg of body weight. A potent ACE inhibitory peptide with the amino acid sequence of Thr-Gln-Val-Tyr (IC50, 18.2 microM) was isolated and identified from the hydrolysate. Single oral administration of Thr-Gln-Val-Tyr at a dose of 30 mg/kg of body weight also significantly decreased blood pressure in SHR. These results suggest that in vitro ACE inhibitory activity and in vivo antihypertensive activity could be generated from rice protein by enzymatic hydrolysis. The rice protein hydrolysate prepared with Alcalase might be utilized to develop physiologically functional food with antihypertensive activity.


Chemical profiling of heroin recovered from the North Korean merchant vessel Pong Su.
Heroin samples, seized from the North Korean merchant vessel Pong Su in Australian waters, were analyzed to determine geographic origin. Duplicate samples were analyzed by the National Measurement Institute's Australian Forensic Drug Laboratory and the United States Drug Enforcement Administration's Special Testing and Research Laboratory. Alkaloid ratios were determined by both liquid chromatography-diode array detection (LC-DAD) and capillary electrophoresis-diode array detection (CE-DAD) techniques. Acid/neutral manufacturing by-products were determined by solvent extraction followed by gas chromatography-mass spectrometry (GC-MS). Solvents, trapped in the heroin particles during manufacture, were detected by both static headspace GC-MS and purge and trap GC-MS. The alkaloid ratios obtained were consistent with heroin of a Southeast Asian (SEA) origin and principal component analysis of the alkaloid results demonstrated the presence of at least four subgroupings within the seizure. The solvent analysis detected diethyl ether and ethyl acetate, solvents typically seen in SEA heroin. However, the acid/neutral analysis revealed compounds not normally seen in heroin of a SEA origin. Furthermore, sterol-like molecules, always detected in the acid/neutral analysis of SEA heroin, were absent from the Pong Su samples. The Pong Su heroin, although similar to SEA heroin, has sufficient differences to classify it as having an unknown origin at the time of this writing.


The Modified Resisted Internal Rotation Test for Detection of Gluteal Tendon Tears.
Greater trochanteric pain syndrome (GTPS) has received increasing attention in recent years. Most patients with GTPS present with trochanteric bursitis and respond to nonoperative treatment. However, a subset of patients may have persistent lateral hip pain or recalcitrant GTPS resulting from an undiagnosed gluteal tendon tear. Recalcitrant GTPS may be a debilitating condition in this patient subset. There is a need for an accurate and evidence-based physical examination maneuver to aid in earlier diagnosis of gluteal tendon tears and timely intervention in these patients. Most studies evaluating gluteal tendinopathy fail to assess surgical indications and instead focus on identifying trochanteric bursitis, which may or may not require surgical treatment. The modified resisted internal rotation test has been used in our practice to detect gluteus medius tendon tears in the recalcitrant GTPS patient population. Fundamental anatomic, biomechanical, electromyographic, and clinical data have been reviewed to make this an evidence-based clinical test for early detection of this pathology.


Role of CpSUB1, a subtilisin-like protease, in Cryptosporidium parvum infection in vitro.
The apicomplexan parasite Cryptosporidium is a significant cause of diarrheal disease worldwide. Previously, we reported that a Cryptosporidium parvum subtilisin-like serine protease activity with furin-type specificity cleaves gp40/15, a glycoprotein that is proteolytically processed into gp40 and gp15, which are implicated in mediating infection of host cells. Neither the enzyme(s) responsible for the protease activity in C. parvum lysates nor those that process gp40/15 are known. There are no furin or other proprotein convertase genes in the C. parvum genome. However, a gene encoding CpSUB1, a subtilisin-like serine protease, is present. In this study, we cloned the CpSUB1 genomic sequence and expressed and purified the recombinant prodomain. Reverse transcriptase PCR analysis of RNA from C. parvum-infected HCT-8 cells revealed that CpSUB1 is expressed throughout infection in vitro. In immunoblots, antiserum to the recombinant CpSUB1 prodomain revealed two major bands, of approximately 64 kDa and approximately 48 kDa, for C. parvum lysates and proteins "shed" during excystation. In immunofluorescence assays, the antiserum reacted with the apical region of sporozoites and merozoites. The recombinant prodomain inhibited protease activity and processing of recombinant gp40/15 by C. parvum lysates but not by furin. Since prodomains are often selective inhibitors of their cognate enzymes, these results suggest that CpSUB1 may be a likely candidate for the protease activity in C. parvum and for processing of gp40/15. Importantly, the recombinant prodomain inhibited C. parvum infection of HCT-8 cells. These studies indicate that CpSUB1 plays a significant role in infection of host cells by the parasite and suggest that this enzyme may serve as a target for intervention.


Mitochondrial DNA, chloroplast DNA and the origins of development in eukaryotic organisms.
Several proposals have been made to explain the rise of multicellular life forms. An internal environment can be created and controlled, germ cells can be protected in novel structures, and increased organismal size allows a "division of labor" among cell types. These proposals describe advantages of multicellular versus unicellular organisms at levels of organization at or above the individual cell. I focus on a subsequent phase of evolution, when multicellular organisms initiated the process of development that later became the more complex embryonic development found in animals and plants. The advantage here is realized at the level of the mitochondrion and chloroplast. The extreme instability of DNA in mitochondria and chloroplasts has not been widely appreciated even though it was first reported four decades ago. Here, I show that the evolutionary success of multicellular animals and plants can be traced to the protection of organellar DNA. Three stages are envisioned. Sequestration allowed mitochondria and chloroplasts to be placed in "quiet" germ line cells so that their DNA is not exposed to the oxidative stress produced by these organelles in "active" somatic cells. This advantage then provided Opportunity, a period of time during which novel processes arose for signaling within and between cells and (in animals) for cell-cell recognition molecules to evolve. Development then led to the enormous diversity of animals and plants. The potency of a somatic stem cell is its potential to generate cell types other than itself, and this is a systems property. One of the biochemical properties required for stemness to emerge from a population of cells might be the metabolic quiescence that protects organellar DNA from oxidative stress.


Systemic sclerosis and the risk of tuberculosis.
Pulmonary involvement is common in patients with systemic sclerosis (SSc), and this condition causes substantial morbidity and mortality. Disrupted immunity from the disease or associated medication may render such patients subject to tuberculosis (TB) infection. However, the relationship between SSc and TB has not yet been investigated. Using the Taiwan National Health Insurance Research Database, 838 patients with SSc diagnosed in Taiwan during 2000-2006 were identified and followed for emergence of TB infection. Incidence rate ratios (IRR) of TB compared to 8380 randomly selected age-, sex-, and comorbidity-matched controls without SSc were calculated. The Cox proportional hazards model was used for multivariate adjustment to identify independent risk factors for TB infection. The risk of TB infection was higher in the SSc cohort than in controls (IRR 2.81, 95% CI 1.36-5.37; p = 0.004), particularly for pulmonary TB (IRR 2.53, 95% CI 1.08-5.30; p = 0.022). Other independent risk factors for TB infection in patients with SSc were age ≥ 60 years [hazard ratio (HR) 3.52, 95% CI 1.10-11.33; p = 0.035] and pulmonary hypertension (PH; HR 6.06, 95% CI 1.59-23.17; p = 0.008). Mortality did not differ for SSc patients with or without TB. In this nationwide study, the incidence of TB infection was significantly higher among patients with SSc than in controls without SSc. Special care should be taken in managing patients with SSc who are at high risk for TB, especially those aged ≥ 60 years or who also have PH.


Structure-Composition-Property Relationships in Polymeric Amorphous Calcium Phosphate-Based Dental Composites.
Our studies of amorphous calcium phosphate (ACP)-based materials over the last decade have yielded bioactive polymeric composites capable of protecting teeth from demineralization or even regenerating lost tooth mineral. The anti-cariogenic/re-mineralizing potential of these ACP composites originates from their propensity, when exposed to the oral environment, to release in a sustained manner sufficient levels of mineral-forming calcium and phosphate ions to promote formation of stable apatitic tooth mineral. However, the less than optimal ACP filler/resin matrix cohesion, excessive polymerization shrinkage and water sorption of these experimental materials can adversely affect their physicochemical and mechanical properties, and, ultimately, limit their lifespan. This study demonstrates the effects of chemical structure and composition of the methacrylate monomers used to form the matrix phase of composites on degree of vinyl conversion (DVC) and water sorption of both copolymers and composites and the release of mineral ions from the composites. Modification of ACP surface via introducing cations and/or polymers ab initio during filler synthesis failed to yield mechanically improved composites. However, moderate improvement in composite's mechanical stability without compromising its remineralization potential was achieved by silanization and/or milling of ACP filler. Using ethoxylated bisphenol A dimethacrylate or urethane dimethacrylate as base monomers and adding moderate amounts of hydrophilic 2-hydroxyethyl methacrylate or its isomer ethyl-α-hydroxymethacrylate appears to be a promising route to maximize the remineralizing ability of the filler while maintaining high DVC. Exploration of the structure/composition/property relationships of ACP fillers and polymer matrices is complex but essential for achieving a better understanding of the fundamental mechanisms that govern dissolution/re-precipitation of bioactive ACP fillers, and, ultimately, the suitability of the composites for clinical evaluation.


Single-mask microfabrication of aspherical optics using KOH anisotropic etching of Si.
We report on the microfabrication of continuous aspherical optical surfaces with a single-mask process, using anisotropic etching of silicon in a KOH water solution. Precise arbitrary aspherical surfaces with lateral scales on the order of several millimeters and a profile depth on the order of several micrometers were fabricated using this process. We discuss the factors defining the precision of the formed component and the resulting surface quality. We demonstrate 1 mm and 5 mm replicated aspherical phase plates, reproducing defocus, tilt, astigmatism and high-order aberrations. The technology has a potential for serial production of reflective and refractive arbitrary aspherical micro-optical components.


Treating atherosclerosis: the potential of Toll-like receptors as therapeutic targets.
Atherosclerosis is an inflammatory disease with a strong involvement of innate immunity. Toll-like receptors (TLRs) are the best-characterized pattern recognition receptors of the innate immune system. Almost all cell types in lesions, inflammatory leukocytes and resident vascular cells alike express TLRs. TLRs are able to sense modified lipids, enhance foam cell formation, induce leukocyte recruitment, and increase cytokine and matrix metalloproteinase production within atherosclerotic lesions. As such, TLRs represent an important link between atheroma and inflammation, making them attractive targets for the treatment of cardiovascular disease. Novel TLR-specific biologics are being developed and tested in other inflammatory diseases. This article will describe the exciting potential of TLRs as therapeutic targets for the treatment of atherosclerosis and will also highlight the potential challenges in the clinical application of TLR-based therapeutics in cardiovascular disease.


Representation of coupled adiabatic potential energy surfaces using neural network based quasi-diabatic Hamiltonians: 1,2 2A' states of LiFH.
An analytic quasi-diabatic representation of ab initio electronic structure data is key to the accurate quantum mechanical description of non-adiabatic chemical processes. In this work, a general neural network (NN) fitting procedure is proposed to generate coupled quasi-diabatic Hamiltonians (Hd) that are capable of representing adiabatic energies, energy gradients, and derivative couplings over a wide range of geometries. The quasi-diabatic representation for LiFH is used as a testing example. The fitting data including adiabatic energies, energy gradients and interstate couplings are obtained from a previously fitted analytical quasi-diabatic potential energy matrix, and are well reproduced by the NN fitting. Most importantly, the NN fitting also yields quantum dynamic results that reproduce those on the original LiFH diabatic Hamiltonian, demonstrating the ability of NN to generate highly accurate quasi-diabatic Hamiltonians.


Associations of Protein Intake and Protein Source with Bone Mineral Density and Fracture Risk: A Population-Based Cohort Study.
High dietary protein has been hypothesized to cause lower bone mineral density (BMD) and greater fracture risk. Previous results are conflicting and few studies have assessed potential differences related to differing protein sources. To determine associations between total protein intake, and protein intake by source (dairy, non-dairy animal, plant) with BMD, BMD change, and incident osteoporotic fracture. Prospective cohort study (Canadian Multicentre Osteoporosis Study). Participants/Measures: Protein intake was assessed as percent of total energy intake (TEI) at Year 2 (1997-99) using a food frequency questionnaire (N=6510). Participants were contacted annually to ascertain incident fracture. Total hip and lumbar spine BMD was measured at baseline and Year 5. Analyses were stratified by group (men 25-49 y, men 50+ y, premenopausal women 25-49 y, and postmenopausal women 50+ y) and adjusted for major confounders. Fracture analyses were limited to those 50+ y. Intakes of dairy protein (with adjustment for BMI) were positively associated with total hip BMD among men and women aged 50+ y, and in men aged 25-49. Among adults aged 50+ y, those with protein intakes of <12% TEI (women) and <11% TEI (men) had increased fracture risk compared to those with intakes of 15% TEI. Fracture risk did not significantly change as intake increased above 15% TEI, and was not significantly associated with protein source. In contrast to hypothesized risk of high protein, we found that for adults 50+ y, low protein intake (below 15% TEI) may lead to increased fracture risk. Source of protein was a determinant of BMD, but not fracture risk.


Total Dietary Fats, Fatty Acids, and Omega-3/Omega-6 Ratio as Risk Factors of Breast Cancer in the Polish Population - a Case-Control Study.
Breast cancer is the most common type of cancer among women around the world and the leading cause of cancer-related death among women. The knowledge about modifiable risk factors, such as diet, can be an acceptable, cheap and non-pharmacological prevention tool. The aim of this study was to investigate the association between dietary fat, dietary fatty acids, fish intake, and breast cancer in women. A case-control study was designed. A total of 201 consecutive, newly diagnosed, polish female cancer patients (mean age: 58 years) and 201 one-to-one age-matched controls were enrolled. A standardised questionnaire assessing various socio-demographic, clinical, lifestyle, and dietary characteristics was applied via face-to-face interviews. Detailed dietary intake information was assessed using a validated Food Frequency Questionnaire. Odds ratios (OR) and 95% confidence intervals (95%CI) were obtained using multiple unconditional logistic regression models controlling for non-dietary and dietary potential confounders. Consumption of polyunsaturated fats (PUFA) over 10% of total energy intake was associated with a significantly lower risk of breast cancer compared to low intake of PUFA (OR=0.4, 95%CI=0.19-0.85). Low (<0.2) omega-3/ omega-6 ratio (OR=2.04, 95%CI=0.996-4.17), fish consumption less than once every six months (OR=3.37, 95%CI=1.57-7.23) and being overweight (OR=2.07, 95%CI=1.3-3.3) were associated with increased risk of breast cancer. Residents of rural areas had a significantly higher risk compared to women from urban areas (OR=1.8, 95%CI=1.06-3.03). High intake of PUFA can decrease the risk of breast cancer, while the low omega-3/omega-6 ratio increases the risk. In addition, overweight state, eliminating fish from the diet and living in rural areas can also increase the risk of breast cancer.


Identification and structure-function analysis of subfamily selective G protein-coupled receptor kinase inhibitors.
Selective inhibitors of individual subfamilies of G protein-coupled receptor kinases (GRKs) would serve as useful chemical probes as well as leads for therapeutic applications ranging from heart failure to Parkinson's disease. To identify such inhibitors, differential scanning fluorimetry was used to screen a collection of known protein kinase inhibitors that could increase the melting points of the two most ubiquitously expressed GRKs: GRK2 and GRK5. Enzymatic assays on 14 of the most stabilizing hits revealed that three exhibit nanomolar potency of inhibition for individual GRKs, some of which exhibiting orders of magnitude selectivity. Most of the identified compounds can be clustered into two chemical classes: indazole/dihydropyrimidine-containing compounds that are selective for GRK2 and pyrrolopyrimidine-containing compounds that potently inhibit GRK1 and GRK5 but with more modest selectivity. The two most potent inhibitors representing each class, GSK180736A and GSK2163632A, were cocrystallized with GRK2 and GRK1, and their atomic structures were determined to 2.6 and 1.85 Å spacings, respectively. GSK180736A, developed as a Rho-associated, coiled-coil-containing protein kinase inhibitor, binds to GRK2 in a manner analogous to that of paroxetine, whereas GSK2163632A, developed as an insulin-like growth factor 1 receptor inhibitor, occupies a novel region of the GRK active site cleft that could likely be exploited to achieve more selectivity. However, neither compound inhibits GRKs more potently than their initial targets. This data provides the foundation for future efforts to rationally design even more potent and selective GRK inhibitors.


Predicting Magnetostimulation Thresholds in the Peripheral Nervous System using Realistic Body Models.
Rapid switching of applied magnetic fields in the kilohertz frequency range in the human body induces electric fields powerful enough to cause Peripheral Nerve Stimulation (PNS). PNS has become one of the main constraints on the use of high gradient fields for fast imaging with the latest MRI gradient technology. In recent MRI gradients, the applied fields are powerful enough that PNS limits their application in fast imaging sequences like echo-planar imaging. Application of Magnetic Particle Imaging (MPI) to humans is similarly PNS constrained. Despite its role as a major constraint, PNS considerations are only indirectly incorporated in the coil design process, mainly through using the size of the linear region as a proxy for PNS thresholds or by conducting human experiments after constructing coil prototypes. We present for the first time, a framework to simulate PNS thresholds for realistic coil geometries to directly address PNS in the design process. Our PNS model consists of an accurate body model for electromagnetic field simulations, an atlas of peripheral nerves, and a neurodynamic model to predict the nerve responses to imposed electric fields. With this model, we were able to reproduce measured PNS thresholds of two leg/arm solenoid coils with good agreement.


Variation in the mineral element concentration of Moringa oleifera Lam. and M. stenopetala (Bak. f.) Cuf.: Role in human nutrition.
Moringa oleifera (MO) and M. stenopetala (MS) (family Moringaceae; order Brassicales) are multipurpose tree/shrub species. They thrive under marginal environmental conditions and produce nutritious edible parts. The aim of this study was to determine the mineral composition of different parts of MO and MS growing in their natural environments and their potential role in alleviating human mineral micronutrient deficiencies (MND) in sub-Saharan Africa. Edible parts of MO (n = 146) and MS (n = 50), co-occurring cereals/vegetables and soils (n = 95) underneath their canopy were sampled from localities in southern Ethiopia and Kenya. The concentrations of seven mineral elements, namely, calcium (Ca), copper (Cu), iodine (I), iron (Fe), magnesium (Mg), selenium (Se), and zinc (Zn) in edible parts and soils were determined using inductively coupled plasma-mass spectrometry. In Ethiopian crops, MS leaves contained the highest median concentrations of all elements except Cu and Zn, which were greater in Enset (a.k.a., false banana). In Kenya, Mo flowers and MS leaves had the highest median Se concentration of 1.56 mg kg-1 and 3.96 mg kg-1, respectively. The median concentration of Se in MS leaves was 7-fold, 10-fold, 23-fold, 117-fold and 147-fold more than that in brassica leaves, amaranth leaves, baobab fruits, sorghum grain and maize grain, respectively. The median Se concentration was 78-fold and 98-fold greater in MO seeds than in sorghum and maize grain, respectively. There was a strong relationship between soil total Se and potassium dihydrogen phosphate (KH2PO4)-extractable Se, and Se concentration in the leaves of MO and MS. This study confirms previous studies that Moringa is a good source of several of the measured mineral nutrients, and it includes the first wide assessment of Se and I concentrations in edible parts of MO and MS grown in various localities. Increasing the consumption of MO and MS, especially the leaves as a fresh vegetable or in powdered form, could reduce the prevalence of MNDs, most notably Se deficiency.


The Genome of the Netherlands: design, and project goals.
Within the Netherlands a national network of biobanks has been established (Biobanking and Biomolecular Research Infrastructure-Netherlands (BBMRI-NL)) as a national node of the European BBMRI. One of the aims of BBMRI-NL is to enrich biobanks with different types of molecular and phenotype data. Here, we describe the Genome of the Netherlands (GoNL), one of the projects within BBMRI-NL. GoNL is a whole-genome-sequencing project in a representative sample consisting of 250 trio-families from all provinces in the Netherlands, which aims to characterize DNA sequence variation in the Dutch population. The parent-offspring trios include adult individuals ranging in age from 19 to 87 years (mean=53 years; SD=16 years) from birth cohorts 1910-1994. Sequencing was done on blood-derived DNA from uncultured cells and accomplished coverage was 14-15x. The family-based design represents a unique resource to assess the frequency of regional variants, accurately reconstruct haplotypes by family-based phasing, characterize short indels and complex structural variants, and establish the rate of de novo mutational events. GoNL will also serve as a reference panel for imputation in the available genome-wide association studies in Dutch and other cohorts to refine association signals and uncover population-specific variants. GoNL will create a catalog of human genetic variation in this sample that is uniquely characterized with respect to micro-geographic location and a wide range of phenotypes. The resource will be made available to the research and medical community to guide the interpretation of sequencing projects. The present paper summarizes the global characteristics of the project.


Molecular High-Grade B-Cell Lymphoma: Defining a Poor-Risk Group That Requires Different Approaches to Therapy.
Biologic heterogeneity is a feature of diffuse large B-cell lymphoma (DLBCL), and the existence of a subgroup with poor prognosis and phenotypic proximity to Burkitt lymphoma is well known. Conventional cytogenetics identifies some patients with rearrangements of MYC and BCL2 and/or BCL6 (double-hit lymphomas) who are increasingly treated with more intensive chemotherapy, but a more biologically coherent and clinically useful definition of this group is required. We defined a molecular high-grade (MHG) group by applying a gene expression-based classifier to 928 patients with DLBCL from a clinical trial that investigated the addition of bortezomib to standard rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) therapy. The prognostic significance of MHG was compared with existing biomarkers. We performed targeted sequencing of 70 genes in 400 patients and explored molecular pathology using gene expression signature databases. Findings were validated in an independent data set. The MHG group comprised 83 patients (9%), with 75 in the cell-of-origin germinal center B-cell-like group. MYC rearranged and double-hit groups were strongly over-represented in MHG but comprised only one half of the total. Gene expression analysis revealed a proliferative phenotype with a relationship to centroblasts. Progression-free survival rate at 36 months after R-CHOP in the MHG group was 37% (95% CI, 24% to 55%) compared with 72% (95% CI, 68% to 77%) for others, and an analysis of treatment effects suggested a possible positive effect of bortezomib. Double-hit lymphomas lacking the MHG signature showed no evidence of worse outcome than other germinal center B-cell-like cases. MHG defines a biologically coherent high-grade B-cell lymphoma group with distinct molecular features and clinical outcomes that effectively doubles the size of the poor-prognosis, double-hit group. Patients with MHG may benefit from intensified chemotherapy or novel targeted therapies.


Selective cleavage of nuclear autoantigens during CD95 (Fas/APO-1)-mediated T cell apoptosis.
Intracellular proteases appear to be important mediators of apoptosis. Substrates cleaved by proteases during apoptosis include nuclear autoantigens targeted in systemic autoimmune diseases. Using human autoantibodies as probes, we demonstrate here that T cell apoptosis mediated by CD95 (Fas/APO-1) is associated with substantial cleavage of a subset of nuclear autoantigens (7 of 33 examined). This subset included poly (ADP-ribose) polymerase, the 70-kD protein of the U1 small nuclear ribonucleoprotein particle, lamin B, the nuclear mitotic apparatus protein NuMA, DNA topoisomerases I and II, and the RNA polymerase I upstream binding factor UBF. Several of the cleaved autoantigens are involved in ensuring the integrity and proper conformation of DNA in the nucleus through interactions with the nuclear matrix, suggesting the possibility that their cleavage may contribute to the collapse of nuclear structure during apoptosis. The relative cleavage kinetics indicated that the autoantigens were targeted at various times after induction of apoptosis, suggesting either differential accessibility or activation of distinct proteases during the cell death process. These data reinforce the hypothesis that apoptosis is accompanied by selective cleavage of key substrates and not by a generalized degradation of intracellular material.


Ca2+ signals during early lymphocyte activation in carp Cyprinus carpio L.
To measure cellular responses and the involvement of increased cytosolic Ca2+ levels ([Ca2+]i), peripheral blood leukocytes (PBL) of carp were loaded with the fluorescent intracellular Ca2+ indicators Fluo-3 and Fura-2. Responses of lymphocytes to T-cell mitogen (phytohaemagglutinin, PHA), to B-cell mitogen (lipopolysaccharide, LPS) and to immunoglobulin (Ig) cross-linking with a monoclonal antibody to carp Ig were measured using flow cytometry. Both T-cell stimulation by PHA and B-cell stimulation by membrane Ig cross-linking evoked a rapid elevation of [Ca2+]i. B-cell stimulation by LPS was not linked to an increase in [Ca2+]i. As judged by the percentage of reacting cells, it was concluded that all Ig-positive lymphocytes reacted to Ig cross-linking by elevating [Ca2+]i. At the single-cell level, the reactions of Fura-2-loaded cells were followed every 6 s using digital imaging microscopy. Both cells displaying spontaneous [Ca2+]i oscillations and non-oscillating cells responded to stimulation with an increase in [Ca2+]i, sometimes, in already oscillating cells, accompanied by an increase in frequency and/or amplitude of the oscillations. These results show that intracellular Ca2+ responses of PBL upon activation resemble those in mammals and form a powerful tool for studies into cell-specific regulation.


Acrosome formation in the house cricket.
Acrosome formation during spermatogenesis in the house cricket was studied with the electron microscope. In the early spermatid there is a single Golgi body, called the acroblast, which is cup-shaped, the walls being composed of a number of parallel membranes. A pro-acrosomal granule then appears within the acroblast. Next, the granule migrates to the nucleus, where it becomes attached. The acroblast then migrates away from the attached granule and is eventually sloughed off. In the first stage of acrosome differentiation the granule assumes the shape of a blunt cone and its base invaginates deeply so that it becomes hollowed. Within the space created by the invagination a new structure forms which, from the first, has the shape of a hollow cone. The two cones constitute the mature acrosome. Both have a biconvex cross-section.