Generation of picoliter droplets with defined contents and concentration gradients from the separation of chemical mixtures.
There has been an increasing drive toward miniaturizing and accelerating experiments with droplet-based microfluidics across the chemical disciplines. Current applications take advantage of the numerous techniques for manipulating nano- to femtoliter droplets within microfluidic devices. To expand the range of possible applications, we have developed a method for compartmentalizing pure compounds within droplets, at a gradient of concentrations, starting from chemical mixtures. In this technique, a mixture is injected into an ultra performance liquid chromatography (UPLC) system, and droplets are generated from the LC output at a frequency high enough to fraction each compound into approximately 10(5) droplets, compartmentalizing pure compounds into a sequence of droplets with a range of concentrations spanning 2-3 orders of magnitude. Here we used fluorescent dyes to quantify the concentration profile of the droplet collections, and to demonstrate the correspondence between the concentration profile of the droplets and the compound elution profile monitored with a UV absorbance detector, allowing the use of compounds that are not fluorescently labeled but show UV absorbance. Hence this technique is applicable to a wide variety of applications that require both compound purity and the ability to probe a variety of concentrations, such as drug screening and titrations.


Medical Informatics Training at Stanford University School of Medicine.
Stanford University School of Medicine has offered graduate degrees in medical informatics since 1982. Located approximately 50 kilometers south of San Francisco near the city of Palo Alto, the university offers both MS and PhD degrees, combining research training with formal course requirements in clinical information sciences, bioinformatics, computer science, decision science, basic biomedicine, health economics, and social and ethical issues. Requirements are adapted to the varying backgrounds of trainees. Graduates of the program work in a variety of capacities, although the majority have sought careers in academia or in industrial research settings.


Poly(alkylidenimine) Dendrimers Functionalized with the Organometallic Moiety [Ru(η⁵-C₅H₅)(PPh₃)₂]⁺ as Promising Drugs Against Cisplatin-Resistant Cancer Cells and Human Mesenchymal Stem Cells.
Here and for the first time, we show that the organometallic compound [Ru(η⁵-C₅H₅)(PPh₃)₂Cl] (RuCp) has potential to be used as a metallodrug in anticancer therapy, and further present a new approach for the cellular delivery of the [Ru(η⁵-C₅H₅)(PPh₃)₂]⁺ fragment via coordination on the periphery of low-generation poly(alkylidenimine) dendrimers through nitrile terminal groups. Importantly, both the RuCp and the dendrimers functionalized with [Ru(η⁵-C₅H₅)(PPh₃)₂]⁺ fragments present remarkable toxicity towards a wide set of cancer cells (Caco-2, MCF-7, CAL-72, and A2780 cells), including cisplatin-resistant human ovarian carcinoma cell lines (A2780cisR cells). Also, RuCp and the prepared metallodendrimers are active against human mesenchymal stem cells (hMSCs), which are often found in the tumor microenvironment where they seem to play a role in tumor progression and drug resistance.


Dendrimer-encapsulated naphthalocyanine as a single agent-based theranostic nanoplatform for near-infrared fluorescence imaging and combinatorial anticancer phototherapy.
Multifunctional theranostic platforms capable of concurrent near-infrared (NIR) fluorescence imaging and phototherapies are strongly desired for cancer diagnosis and treatment. However, the integration of separate imaging and therapeutic components into nanocarriers results in complex theranostic systems with limited translational potential. A single agent-based theranostic nanoplatform, therefore, was developed for concurrent NIR fluorescence imaging and combinatorial phototherapy with dual photodynamic (PDT) and photothermal (PTT) therapeutic mechanisms. The transformation of a substituted silicon naphthalocyanine (SiNc) into a biocompatible nanoplatform (SiNc-NP) was achieved by SiNc encapsulation into the hydrophobic interior of a generation 5 polypropylenimine dendrimer following surface modification with polyethylene glycol. Encapsulation provides aqueous solubility to SiNc and preserves its NIR fluorescence, PDT and PTT properties. Moreover, an impressive photostability in the dendrimer-encapsulated SiNc has been detected. Under NIR irradiation (785 nm, 1.3 W cm(-2)), SiNc-NP manifested robust heat generation capability (ΔT = 40 °C) and efficiently produced reactive oxygen species essential for PTT and PDT, respectively, without releasing SiNc from the nanopaltform. By varying the laser power density from 0.3 W cm(-2) to 1.3 W cm(-2) the therapeutic mechanism of SiNc-NP could be switched from PDT to combinatorial PDT-PTT treatment. In vitro and in vivo studies confirmed that phototherapy mediated by SiNc can efficiently destroy chemotherapy resistant ovarian cancer cells. Remarkably, solid tumors treated with a single dose of SiNc-NP combined with NIR irradiation were completely eradicated without cancer recurrence. Finally, the efficiency of SiNc-NP as an NIR imaging agent was confirmed by recording the strong fluorescence signal in the tumor, which was not photobleached during the phototherapeutic procedure.


Tanycytic networks mediate energy balance by feeding lactate to glucose-insensitive POMC neurons.
Hypothalamic glucose sensing enables an organism to match energy expenditure and food intake to circulating levels of glucose, the main energy source of the brain. Here, we established that tanycytes of the arcuate nucleus of the hypothalamus, specialized glia that line the wall of the third ventricle, convert brain glucose supplies into lactate that they transmit through monocarboxylate transporters to arcuate proopiomelanocortin neurons, which integrate this signal to drive their activity and to adapt the metabolic response to meet physiological demands. Furthermore, this transmission required the formation of extensive connexin-43 gap junction-mediated metabolic networks by arcuate tanycytes. Selective suppression of either tanycytic monocarboxylate transporters or gap junctions resulted in altered feeding behavior and energy metabolism. Tanycytic intercellular communication and lactate production are thus integral to the mechanism by which hypothalamic neurons that regulate energy and glucose homeostasis efficiently perceive alterations in systemic glucose levels as a function of the physiological state of the organism.


Impact of chemotherapy on telomere length in sporadic and familial breast cancer patients.
Recently, we observed that telomeres of BRCA1/2 mutation carriers were shorter than those of controls or sporadic breast cancer patients, suggesting that mutations in these genes might be responsible for this event. Given the contradictory results reported in the literature, we tested whether other parameters, such as chemotherapy, could be modifying telomere length (TL). We performed a cross-sectional study measuring leukocyte TL of 266 sporadic breasts cancer patients treated with first-line chemotherapy, with a median follow-up of 240 days. Additionally, we performed both cross-sectional and longitudinal studies in a series of 236 familial breast cancer patients that included affected and non-affected BRCA1/2 mutation carriers. We have measured in leukocytes from peripheral blood: the TL, percentage of short telomeres (<3 kb), telomerase activity levels and the annual telomere shortening speed. In sporadic cases we found that chemotherapy exerts a transient telomere shortening effect (around 2 years) that varies depending on the drug combination. In familial cases, only patients receiving treatment were associated with telomere shortening but they recovered normal TL after a period of 2 years. Chemotherapy affects TL and should be considered in the studies that correlate TL with disease susceptibility.


A diabetes dashboard and physician efficiency and accuracy in accessing data needed for high-quality diabetes care.
We compared use of a new diabetes dashboard screen with use of a conventional approach of viewing multiple electronic health record (EHR) screens to find data needed for ambulatory diabetes care. We performed a usability study, including a quantitative time study and qualitative analysis of information-seeking behaviors. While being recorded with Morae Recorder software and "think-aloud" interview methods, 10 primary care physicians first searched their EHR for 10 diabetes data elements using a conventional approach for a simulated patient, and then using a new diabetes dashboard for another. We measured time, number of mouse clicks, and accuracy. Two coders analyzed think-aloud and interview data using grounded theory methodology. The mean time needed to find all data elements was 5.5 minutes using the conventional approach vs 1.3 minutes using the diabetes dashboard (P <.001). Physicians correctly identified 94% of the data requested using the conventional method, vs 100% with the dashboard (P <.01). The mean number of mouse clicks was 60 for conventional searching vs 3 clicks with the diabetes dashboard (P <.001). A common theme was that in everyday practice, if physicians had to spend too much time searching for data, they would either continue without it or order a test again. Using a patient-specific diabetes dashboard improves both the efficiency and accuracy of acquiring data needed for high-quality diabetes care. Usability analysis tools can provide important insights into the value of optimizing physician use of health information technologies.


Is metal accumulation in Pomphorhynchus laevis dependent on parasite sex or infrapopulation size?
Concentrations of the elements As, Cd, Co, Cu, Fe, Mn, Mo, Ni, Pb, Sn, V, Zn were analysed by inductively coupled plasma mass spectrometry (ICP-MS) in the acanthocephalan Pomphorhynchus laevis and its fish host Barbus barbus. A total of 27 barbels were collected from the Danube River in autumn 2006 close to the town Kozloduy (685 river kilometer) on the Bulgarian river bank. Fish were divided into 3 groups. According to their P. laevis infrapopulation size hosts were considered as heavily infected (>100 worms per fish) and lightly infected (<20 worms per fish). The third group was used to compare heavy metal concentrations between male and female P. laevis. The 5 elements As, Cd, Cu, Pb and Zn were detected in significantly higher concentrations in parasites compared to host tissues (muscle, intestine, liver). According to the calculated mean bioconcentration factors, 3 more elements (Co, Mn, V) showed usually higher concentrations in P. laevis. Comparisons between heavily and lightly infected fish revealed significant differences only for V with higher concentrations for the heavily infected group. Concerning sex-specific metal accumulation V and Zn showed significant differences (V, at P<0.05; Zn, at P=0.05), with higher levels of both metals in females of P. laevis. Our results suggest that - for the metals analysed - the size of the parasite infrapopulation plays no role in the degree of metal accumulation. Similarly, parasite sex seems not to be a crucial factor for metal accumulation in the parasites. Thus, for metal monitoring purposes there is no need to take these aspects into account, which makes the use of parasites as bioindicators more applicable.


Pressure ulcers in trauma patients with suspected spine injury: a prospective cohort study with emphasis on device-related pressure ulcers.
Of all patients in a hospital environment, trauma patients may be particularly at risk for developing (device-related) pressure ulcers (PUs), because of their traumatic injuries, immobility, and exposure to immobilizing and medical devices. Studies on device-related PUs are scarce. With this study, the incidence and characteristics of PUs and the proportion of PUs that are related to devices in adult trauma patients with suspected spinal injury were described. From January-December 2013, 254 trauma patients were visited every 2 days for skin assessment. The overall incidence of PUs was 28·3% (n = 72/254 patients). The incidence of device-related PUs was 20·1% (n = 51), and 13% (n = 33) developed solely device-related PUs. We observed 145 PUs in total of which 60·7% were related to devices (88/145). Device-related PUs were detected 16 different locations on the front and back of the body. These results show that the incidence of PUs and the proportion of device-related PUs is very high in trauma patients.


In defense of causal-formative indicators: A minority report.
Causal-formative indicators directly affect their corresponding latent variable. They run counter to the predominant view that indicators depend on latent variables and are thus often controversial. If present, such indicators have serious implications for factor analysis, reliability theory, item response theory, structural equation models, and most measurement approaches that are based on reflective or effect indicators. Psychological Methods has published a number of influential articles on causal and formative indicators as well as launching the first major backlash against them. This article examines 7 common criticisms of these indicators distilled from the literature: (a) A construct measured with "formative" indicators does not exist independently of its indicators; (b) Such indicators are causes rather than measures; (c) They imply multiple dimensions to a construct and this is a liability; (d) They are assumed to be error-free, which is unrealistic; (e) They are inherently subject to interpretational confounding; (f) They fail proportionality constraints; and (g) Their coefficients should be set in advance and not estimated. We summarize each of these criticisms and point out the flaws in the logic and evidence marshaled in their support. The most common problems are not distinguishing between what we call causal-formative and composite-formative indicators, tautological fallacies, and highlighting issues that are common to all indicators, but presenting them as special problems of causal-formative indicators. We conclude that measurement theory needs (a) to incorporate these types of indicators, and (b) to better understand their similarities to and differences from traditional indicators. (PsycINFO Database Record


Weight Gain and Its Correlates Among Forensic Inpatients.
We investigated changes in weight, body mass index (BMI), and other indices of the metabolic syndrome in forensic inpatients. Weight gain associated with newer antipsychotics (APs) is well established in the general psychiatric population. We examined the medical records of 291 men admitted to a forensic hospital at admission and again at discharge or 365 days later if still in hospital. We also recorded diagnosis and smoker status on admission and quantified psychotropic treatment and adherence, physical activity, and daytime occupation during the hospitalization. On admission, 33% were obese and 22% of the 106 patients for whom sufficient data were available met criteria for metabolic syndrome. Among patients staying at least 30 days, 60% were weighed again before discharge but repeated blood pressure and waist circumference measures were uncommon, even among those at greatest risk. The 122 forensic inpatients with sufficient information gained an average of 12% of their body weight and 40% increased by at least 1 BMI category, gaining an average of 3.67 kg per month. Weight gain was associated with duration of time and was not attributable to being underweight on admission, diagnosis of schizophrenia, atypical AP treatment, medication adherence, or having been a smoker. Patients gained weight during forensic hospitalization independent of medication use. We recommend further research using consistent measurement and wider sampling of both metabolic syndrome indicators and its individual and systemic causes in forensic populations.


Common and differential connectivity profiles of deep brain stimulation and capsulotomy in refractory obsessive-compulsive disorder.
Neurosurgical interventions including deep brain stimulation (DBS) and capsulotomy have been demonstrated effective for refractory obsessive-compulsive disorder (OCD), although treatment-shared/-specific network mechanisms remain largely unclear. We retrospectively analyzed resting-state fMRI data from three cohorts: a cross-sectional dataset of 186 subjects (104 OCD and 82 healthy controls), and two longitudinal datasets of refractory patients receiving ventral capsule/ventral striatum DBS (14 OCD) and anterior capsulotomy (27 OCD). We developed a machine learning model predictive of OCD symptoms (indexed by the Yale-Brown Obsessive Compulsive Scale, Y-BOCS) based on functional connectivity profiles and used graphic measures of network communication to characterize treatment-induced profile changes. We applied a linear model on 2 levels treatments (DBS or capsulotomy) and outcome to identify whether pre-surgical network communication was associated with differential treatment outcomes. We identified 54 functional connectivities within fronto-subcortical networks significantly predictive of Y-BOCS score in patients across 3 independent cohorts, and observed a coexisting pattern of downregulated cortico-subcortical and upregulated cortico-cortical network communication commonly shared by DBS and capsulotomy. Furthermore, increased cortico-cortical communication at ventrolateral and centrolateral prefrontal cortices induced by DBS and capsulotomy contributed to improvement of mood and anxiety symptoms, respectively (p < 0.05). Importantly, pretreatment communication of ventrolateral and centrolateral prefrontal cortices were differentially predictive of mood and anxiety improvements by DBS and capsulotomy (effect sizes = 0.45 and 0.41, respectively). These findings unravel treatment-shared and treatment-specific network characteristics induced by DBS and capsulotomy, which may facilitate the search of potential evidence-based markers for optimally selecting among treatment options for a patient.


FEEL: a system for frequent event and electrodermal activity labeling.
The wide availability of low-cost wearable biophysiological sensors enables us to measure how the environment and our experiences impact our physiology. This creates a challenge: in order to interpret the longitudinal data, we require the matching contextual information as well. Collecting continuous biophysiological data makes it unfeasible to rely solely on our memory for contextual information. In this paper, we first present an architecture and implementation of a system for the acquisition, processing, and visualization of biophysiological signals and contextual information. Next, we present the results of a user study: users wore electrodermal activity wrist sensors that measured their autonomic arousal. These users uploaded the sensor data at the end of each day. At first, they annotated their events at the end of each day; then, after a two-day break, they annotated the data from two days earlier. One group of users had access to both the signal and the contextual information collected by the mobile phone and the other group could only access the biophysiological signal. At the end of the study, the users filled in a system usability scale and user experience surveys. Our results show that the system enables the users to annotate biophysiological signals at a greater effectiveness than the current state of the art while also providing very good usability.


Rapid, activity-independent turnover of vesicular transmitter content at a mixed glycine/GABA synapse.
The release of neurotransmitter via the fusion of transmitter-filled, presynaptic vesicles is the primary means by which neurons relay information. However, little is known regarding the molecular mechanisms that supply neurotransmitter destined for vesicle filling, the endogenous transmitter concentrations inside presynaptic nerve terminals, or the dynamics of vesicle refilling after exocytosis. We addressed these issues by recording from synaptically coupled pairs of glycine/GABA coreleasing interneurons (cartwheel cells) of the mouse dorsal cochlear nucleus. We find that the plasma membrane transporter GlyT2 and the intracellular enzyme glutamate decarboxylase supply the majority of glycine and GABA, respectively. Pharmacological block of GlyT2 or glutamate decarboxylase led to rapid and complete rundown of transmission, whereas increasing GABA synthesis via intracellular glutamate uncaging dramatically potentiated GABA release within 1 min. These effects were surprisingly independent of exocytosis, indicating that prefilled vesicles re-equilibrated upon acute changes in cytosolic transmitter. Titration of cytosolic transmitter with postsynaptic responses indicated that endogenous, nonvesicular glycine/GABA levels in nerve terminals are 5-7 mm, and that vesicular transport mechanisms are not saturated under basal conditions. Thus, cytosolic transmitter levels dynamically set the strength of inhibitory synapses in a release-independent manner.


Transcription factor Sp4 regulates expression of nervous wreck 2 to control NMDAR1 levels and dendrite patterning.
Glutamatergic signaling through N-methyl-d-aspartate receptors (NMDARs) is important for neuronal development and plasticity and is often dysregulated in psychiatric disorders. Mice mutant for the transcription factor Sp4 have reduced levels of NMDAR subunit 1 (NR1) protein, but not mRNA, and exhibit behavioral and memory deficits (Zhou et al., [2010] Human Molecular Genetics 19: 3797-3805). In developing cerebellar granule neurons (CGNs), Sp4 controls dendrite patterning (Ramos et al., [2007] Proc Natl Acad Sci USA 104: 9882-9887). Sp4 target genes that regulate dendrite pruning or NR1 levels are not known. Here we report that Sp4 activates transcription of Nervous Wreck 2 (Nwk2; also known as Fchsd1) and, further, that Nwk2, an F-BAR domain-containing protein, mediates Sp4-dependent regulation of dendrite patterning and cell surface expression of NR1. Knockdown of Nwk2 in CGNs increased primary dendrite number, phenocopying Sp4 knockdown, and exogenous expression of Nwk2 in Sp4-depleted neurons rescued dendrite number. We observed that acute Sp4 depletion reduced levels of surface, but not total, NR1, and this was rescued by Nwk2 expression. Furthermore, expression of Nr1 suppressed the increase in dendrite number in Sp4- or Nwk2- depleted neurons. We previously reported that Sp4 protein levels were reduced in cerebellum of subjects with bipolar disorder (BD) (Pinacho et al., [2011] Bipolar Disorders 13: 474-485). Here we report that Nwk2 mRNA and NR1 protein levels were also reduced in postmortem cerebellum of BD subjects. Our data suggest a role for Sp4-regulated Nwk2 in NMDAR trafficking and identify a Sp4-Nwk2-NMDAR1 pathway that regulates neuronal morphogenesis during development and may be disrupted in bipolar disorder.


Toll-like receptor 2 senses hepatitis C virus core protein but not infectious viral particles.
Toll-like receptors (TLRs) are pathogen recognition molecules activating the innate immune system. Cell surface expressed TLRs, such as TLR2 and TLR4, have been shown to play an important role in human host defenses against viruses through sensing of viral structural proteins. In this study, we aimed to elucidate whether TLR2 and TLR4 participate in inducing antiviral immunity against hepatitis C virus (HCV) by sensing viral structural proteins. We studied TLR2 and TLR4 activation by cell culture-derived infectious virions and serum-derived virions in comparison to purified recombinant HCV structural proteins and enveloped virus-like particles. Incubation of TLR2 or TLR4 transfected cell lines with recombinant core protein resulted in activation of TLR2-dependent signaling. In contrast, neither infectious virions nor enveloped HCV-like particles triggered TLR2 and TLR4 signaling. These findings suggest that monomeric HCV core protein but not intact infectious particles are sensed by TLR2. Impairment of interaction between TLR and the core in infectious viral particles may contribute to escape from innate antiviral immune responses.


Clarifying values and transforming attitudes to improve access to second trimester abortion.
Access to safe second trimester abortion services is poor in many countries, sometimes despite liberal laws and policies. Addressing the myriad factors hindering access to safe abortion care requires a multi-pronged strategy. Workshops aimed at clarifying values are useful for addressing barriers to access stemming from misinformation, stigmatization of women and providers, and negative attitudes and obstructionist behaviours. They engage health care providers and administrators, policymakers, community members and others in a process of self-examination with the goal of transforming abortion-related attitudes and behaviours in a direction supportive of women seeking abortion. This is especially important for women seeking second trimester abortion, which tends to be even more stigmatized than first trimester abortion. This paper reports on some promising experiences and results from workshops in Viet Nam, Nepal and South Africa. Some recommendations that emerge are that values clarification should be included in abortion training, service delivery and advocacy programmes. Evaluations of such interventions are also needed.


Associations of mental distress with residency in conflict zones, ethnic minority status, and potentially modifiable social factors following conflict in Sri Lanka: a nationwide cross-sectional study.
The subject of post-conflict mental health lacks studies that are nationally representative or large enough to allow robust examination of levels of distress according to residency in geographical zones of conflict and ethnic minority status. We undertook a nationwide study in Sri Lanka to address these issues. We used tablet devices to survey 20,632 people across 18 of 25 districts in Sri Lanka, of which nine were purposefully selected and nine randomly selected districts. Based on their exposure to war and levels of population displacement, these districts were classified as being in the severe, moderate, or minimal conflict zones. Trained local fieldworkers did private interviews with the participants in the local language; responses were entered directly into the tablet devices. The questions assessed exposure to a wide range of stresses, including ongoing adversities, threat or protection issues, and service access factors, and respondents provided categorical responses (yes/no) to a list of items. We used the 25-item Hopkins Symptom Checklist to measure depression and anxiety in participants. We used multivariate analysis and calculated population-attributable fractions to estimate potential improvement in mental distress if modifiable factors were addressed. Our results showed a stepwise increase in symptoms of depression (10%, 33%, 40%) and anxiety (13%, 23%, 23%) across minimal, moderate, and severe conflict zones, respectively. Membership of an ethnic minority group was associated with depression (Tamil odds ratio [OR] 2·4 [95% CI 1·8-3·1], other ethnic minority OR 2·7 [2·1-3·6]) and anxiety (Tamil OR 1·5 [1·1-2·0], other ethnic minority OR 1·5 [1·2-1·8]). Other fixed characteristics associated with depression and anxiety were older age (both depression and anxiety; OR 2·7 [95% CI 2·0-3·7] for anxiety and 2·3 [1·7-3·0] for depression), being married (anxiety only; OR 1·5 [95% CI 1·1-2·1]), and time spent in camps for internally displaced people (anxiety only; OR 1·4 [95% CI 1·0-1·9]). Modifiable characteristics were threat or protection issues (depression OR 1·8 [95% CI 1·2-2·8], anxiety OR 1·8 [1·3-2·6]) and scarcity of food (depression OR 1·4 [1·2-1·6], anxiety OR 1·4 [1·2-2·7]), loss of job (OR 1·2 [95% CI 1·2-1·4] for anxiety), and proximity to army camps (OR 1·6 [95% CI 1·2-2·2] for depression). Allaying of safety concerns had the potential to reduce depression by 53% and anxiety by 57%, with other factors (removal of army camps from proximity and addressing food scarcity, loss of jobs and threat/protection issues) making smaller contributions to a reduction in depression and anxiety. Our findings offer a basis for detecting high-risk groups and provide directions in devising policies and programmes aimed at alleviating psychological distress in Sri Lanka. Restoration of safety seems to be crucial to mental health recovery. Australian National University-Department of Immigration Border Protection Collaborative Research Program.


Paediatric abdominal tuberculosis in developed countries: case series and literature review.
To provide an insight into the presentation, diagnosis and management of paediatric abdominal tuberculosis (TB) in developed countries. The records of all children at the Montreal Children's Hospital (MCH) admitted with abdominal TB between 1990 and 2014 were reviewed. An automated and manual literature search from 1946 to 2014 was performed. (1) CASE SERIES: six cases were identified at the MCH. All were male between 5 and 17 years of age. All were from populations known to have high rates of TB (aboriginal, immigrant). Three underwent major surgical interventions and three underwent ultrasound (US) or CT aspiration or biopsy for diagnosis. (2) LITERATURE REVIEW: 29 male (64%) and 16 female subjects (36%) aged between 14 months and 18 years were identified, including the MCH patients. All patients except one were from populations with a high incidence of TB. Most presented with a positive tuberculin skin test (90%), abdominal pain (76%), fever (71%) and weight loss (68%). On imaging, 22 (49%) were classified with gastrointestinal TB with colonic wall irregularity (41%) and 19 (42%) with peritoneal TB with ascites (68%). A positive culture was obtained in 33 (73%) patients. Three cases used CT- or US-guided aspiration or biopsy to obtain tissue samples. A surgical intervention was performed in 34 (76%) children; 13 (38%) of these were for diagnosis. Diagnosis based on clinical features (abdominal pain, fever and weight loss) and CT- or US-guided aspiration or biopsy may encourage physicians to adopt a more conservative approach to abdominal TB.


Intermethod reliability of real-time versus delayed videotaped evaluation of a high-fidelity medical simulation septic shock scenario.
High-fidelity medical simulation (HFMS) is increasingly utilized in resident education and evaluation. No criterion standard of assessing performance currently exists. This study compared the intermethod reliability of real-time versus videotaped evaluation of HFMS participant performance. Twenty-five emergency medicine residents and one transitional resident participated in a septic shock HFMS scenario. Four evaluators assessed the performance of participants on technical (26-item yes/no completion) and nontechnical (seven item, five-point Likert scale assessment) scorecards. Two evaluators provided assessment in real time, and two provided delayed videotape review. After 13 scenarios, evaluators crossed over and completed the scenarios in the opposite method. Real-time evaluations were completed immediately at the end of the simulation; videotape reviewers were allowed to review the scenarios with no time limit. Agreement between raters was tested using the intraclass correlation coefficient (ICC), with Cronbach's alpha used to measure consistency among items on the scores on the checklists. Bland-Altman plot analysis of both conditions revealed substantial agreement between the real-time and videotaped review scores by reviewers. The mean difference between the reviewers was 0.0 (95% confidence interval [CI] = -3.7 to 3.6) on the technical evaluation and -1.6 (95% CI = -11.4 to 8.2) on the nontechnical scorecard assessment. Comparison of evaluations for the videotape technical scorecard demonstrated a Cronbach's alpha of 0.914, with an ICC of 0.842 (95% CI = 0.679 to 0.926), and the real-time technical scorecard demonstrated a Cronbach's alpha of 0.899, with an ICC of 0.817 (95% CI = 0.633 to 0.914), demonstrating excellent intermethod reliability. Comparison of evaluations for the videotape nontechnical scorecard demonstrated a Cronbach's alpha of 0.888, with an ICC of 0.798 (95% CI = 0.600 to 0.904), and the real-time nontechnical scorecard demonstrated a Cronbach's alpha of 0.833, with an ICC of 0.714 (95% CI = 0.457 to 0.861), demonstrating substantial interrater reliability. The raters were consistent in agreement on performance within each level of training, as the analysis of variance demonstrated no significant differences between the technical scorecard (p = 0.176) and nontechnical scorecard (p = 0.367). Real-time and videotaped-based evaluations of resident performance of both technical and nontechnical skills during an HFMS septic shock scenario provided equally reliable methods of assessment.


Relating structural disorder and melting in complex mixed ligand zeolitic imidazolate framework glasses.
We report the formation of zeolitic imidazolate framework glasses incorporating three organic linkers, from their corresponding novel crystalline structures [Zn(Im2-x-ybImxmbImy)]. Structure-property relationships between chemical compositions and thermal properties are analysed, in addition to the effect on the nanoscale porosity of the glasses formed. A probabilistic model is used to explain melting and the glass transition temperatures of the obtained glasses and link to the nanoscale structural disorder of their crystalline starting structures.


Primary prevention of cardiovascular disease: new guidelines, technologies and therapies.
A trend in primary prevention of cardiovascular disease (CVD) has been a move away from managing isolated risk factors, such as hypertension and dyslipidaemia, towards assessment and management of absolute CVD risk. In Australian guidelines, absolute CVD risk is calculated as the probability of a stroke, transient ischaemic attack, myocardial infarction, angina, peripheral arterial disease or heart failure occurring within the next 5 2013s. Absolute CVD risk should be regularly assessed in patients aged 45 2013s or older (35 2013s or older in Aboriginal and Torres Strait Islander people) using the Australian absolute CVD risk calculator (http://www.cvdcheck.org.au). For patients currently taking a blood pressure (BP)-lowering or lipid-lowering agent, pretreatment values should be used to calculate risk. Patients at high absolute risk of CVD (> 15% over 5 2013s) should be treated with both BP-lowering and lipid-lowering agents, unless contraindicated or clinically inappropriate. For patients at moderate absolute risk of CVD (10%-15%) treatment with a BP-lowering and/or a lipid-lowering agent should be considered if the risk remains elevated after lifestyle interventions, BP is ≥ 160/100 mmHg, there is a family history of premature CVD, or the patient is of South Asian, Middle Eastern, Maori, Pacific Islander, Aboriginal or Torres Strait Islander ethnicity. BP measurements taken using an oscillometric device can be used to approximate mean daytime ambulatory BP.


Side effects of generic competition?
This study examined the relationship between generic drug market shares and the number of reported side effects. Yearly time-series data for the number of reported side effects and information on market shares, prices, and quantities from 1972 to 1996 were used in this study. Poisson and negative binomial regression models were used in the statistical analysis. The results show that increased generic market share increases the number of reported side effects for all estimated models. When studying the relationship at the substance level, increasing generic market shares increases the number of side effects for 7 of the 15 substances. Generic substitution laws and measures to increase generic competition may thus have unintended consequences since these results show a positive relationship between generic market shares and reported side effects.


Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na+, K+-ATPase control of cell adhesion, proliferation, and survival.
The ion pump Na+, K+-ATPase (NKA) is a receptor for the cardiotonic steroid ouabain. Subsaturating concentration of ouabain triggers intracellular calcium oscillations, stimulates cell proliferation and adhesion, and protects from apoptosis. However, it is controversial whether ouabain-bound NKA is considered a signal transducer. To address this question, we performed a global analysis of protein phosphorylation in COS-7 cells, identifying 2580 regulated phosphorylation events on 1242 proteins upon 10- and 20-min treatment with ouabain. Regulated phosphorylated proteins include the inositol triphosphate receptor and stromal interaction molecule, which are essential for initiating calcium oscillations. Hierarchical clustering revealed that ouabain triggers a structured phosphorylation response that occurs in a well-defined, time-dependent manner and affects specific cellular processes, including cell proliferation and cell-cell junctions. We additionally identify regulation of the phosphorylation of several calcium and calmodulin-dependent protein kinases (CAMKs), including 2 sites of CAMK type II-γ (CAMK2G), a protein known to regulate apoptosis. To verify the significance of this result, CAMK2G was knocked down in primary kidney cells. CAMK2G knockdown impaired ouabain-dependent protection from apoptosis upon treatment with high glucose or serum deprivation. In conclusion, we establish NKA as the coordinator of a broad, tightly regulated phosphorylation response in cells and define CAMK2G as a downstream effector of NKA.-Panizza, E., Zhang, L., Fontana, J. M., Hamada, K., Svensson, D., Akkuratov, E. E., Scott, L., Mikoshiba, K., Brismar, H., Lehtiö, J., Aperia, A. Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na+, K+-ATPase control of cell adhesion, proliferation, and survival.


A pilot trial of low-dose naltrexone in primary progressive multiple sclerosis.
A sixth month phase II multicenter-pilot trial with a low dose of the opiate antagonist Naltrexone (LDN) has been carried out in 40 patients with primary progressive multiple sclerosis (PPMS). The primary end points were safety and tolerability. Secondary outcomes were efficacy on spasticity, pain, fatigue, depression, and quality of life. Clinical and biochemical evaluations were serially performed. Protein concentration of beta-endorphins (BE) and mRNA levels and allelic variants of the mu-opiod receptor gene (OPRM1) were analyzed. Five dropouts and two major adverse events occurred. The remaining adverse events did not interfere with daily living. Neurological disability progressed in only one patient. A significant reduction of spasticity was measured at the end of the trial. BE concentration increased during the trial, but no association was found between OPRM1 variants and improvement of spasticity. Our data clearly indicate that LDN is safe and well tolerated in patients with PPMS.


Racial Disparities of Adrenalectomy.
Racial disparities of surgical outcomes have been demonstrated for a variety of operations. We sought to determine whether minority status is associated with disparate care for adrenalectomy. This study is a retrospective database review of the Collaborative Endocrine Surgery Quality Improvement Program (CESQIP) from January 1, 2014 through April 30, 2018. Primary outcome was complication rate. Secondary outcomes were length of stay and surgeon experience. Minority status was defined as Black or Hispanic and outcomes were compared with White patients. For the study period, 1141 patients who underwent adrenalectomies were included, of whom 69.9% were White and 22.6% minority. The minority patients were significantly younger with higher rates of diabetes mellitus. Minority patients had higher rates of complication and longer length of stay. Minority patients were more likely to have an adrenalectomy by a low-volume surgeon. In multivariate logistic regression, minority status remained associated with complication rate. Minority patients undergoing adrenalectomy have higher rates of complication and longer lengths of stay when controlling for common comorbidities. Minority patients have decreased access to high-volume surgeons.


Individual human odor fallout as detected by trained canines.
We tested the hypothesis that if odor fallout (the release of a human's odor onto an untouched object) in human subjects exists, then holding a hand above an absorbent will produce a detectable scent which will be subsequently matched in a detection test by trained canines. Scents were collected from seven males to sterile cotton absorbent squares. The left hand was used to get the control scent and the right hand served as the target scent. Each experimental subject was sitting; his left hand was laid down on a cotton square for 3 min. The right hand was held 5 cm above another cotton square for 3 min. The scent identification was done by two specially trained police German shepherds. These canines had routinely performed scent identification line-ups as part of criminal investigation procedures. Both canines performed 14 line-ups and correctly matched the collected scents of all test subjects. The results suggest the existence of human odor fallout, whereby a human scent trace is left by humans even if they do not touch an object.


Association between social participation and hypertension among older people in Japan: the JAGES Study.
Hypertension is an important risk factor for cardiovascular disease, the leading cause of mortality in the world. Although previous studies have focused on individual-level behavioral risk factors associated with hypertension, there has been little research on how interacting with others, that is social participation, affects hypertension. To address this research gap, this study examined the association between social participation and hypertension in Japan, a country with a high prevalence of hypertension possibly linked to rapid population aging. Data were used from 4582 participants aged more than 65 years who participated in the Japan Gerontological Evaluation Survey (JAGES) with blood pressure data collected during a health check-up. The frequency of participation in vertical organizations (characterized by hierarchical relationships) and horizontal organizations (characterized by non-hierarchical, egalitarian relationships) was measured by a questionnaire. In a Poisson regression analysis, participation in vertical organizations was not associated with hypertension, whereas participation in horizontal organizations at least once a month was inversely associated with hypertension (prevalence ratio: 0.941). This association remained significant after adjusting for social support variables, although further adjustment for health behaviors attenuated the association. As the frequency of going out and average time spent walking were both associated with hypertension, physical activity may be a possible pathway that connects social participation and hypertension. The results of this study suggest that expanding social participation programs, especially those involving horizontal organizations, may be one way to promote better health among older people in Japan.


Making sense of aches and pains.
The uncertainty about the status of upper limb disorders (ULDs), particularly the non-specific conditions, is believed to have consequences for clinical management and patient care. This paper presents evidence about how sufferers with ULDs respond to their pain, how their pain is managed, when and who they go to for formal help and how sufferers evaluate the care they receive. The data analysis is derived from face-to-face, informal interviews with sufferers with a broad spectrum of upper limb disorders (n = 47). These informants were selected according to strict criteria from a 'screening' postal survey of the working population (25-64 years) in south-west England (n = 2781). Ideas about causation were crucial to understanding patterns of illness action and help seeking behaviour. The common strategy was to wait and see what happens as the pain was believed to be a natural part of the ageing process. Explanations invoking psychosocial and work related causes were less common and tended to be used when biomechanical explanations were no longer appropriate. Self-management was the preferred strategy but orthodox practitioners were usually the first choice for formal care. Complementary and alternative medicines (CAM) were popular but were used to complement orthodox care. Practitioners were evaluated mainly in terms of their ability to alleviate pain. There is a need for orthodox and non-orthodox care to be closely integrated in primary care and GPs should not depend on orthodox medications alone when caring for patients with upper limb pain.


Superresolution optical system by common-path interferometry.
We present a new approach to obtain superresolved images in digital holography by means of synthetic aperture generation using common-path interferometry and off-axis illumination in optical imaging systems. The paper includes two parts. First, we present a simple approach to double the resolution of an optical system using tilted illumination onto the object and an optical element in the image plane to produce the holographic recording. Then we present a novel approach consisting of attaching a diffraction grating in parallel together with the object in the input plane and using off-axis illumination provided by a Vertical Cavity Surface Emitting Lasers (VCSEL) array to allow us achieving a major improvement in the optical resolution limit with an extremely low penalty in the complexity of the resulting system. Experimental investigation based on commercial microscope objectives is presented.


Lack of correlation of functional scintigraphy with (99m)technetium-methoxyisobutylisonitrile with histological necrosis following induction chemotherapy or measures of P-glycoprotein expression in high-grade osteosarcoma.
In osteosarcoma, some studies have suggested P-glycoprotein expression is a prognostic factor. The clearance of (99m)technetium hexakis-2-methoxyisobutylisonitrile ((99m)Tc-MIBI) has been used in some tumor systems as an in vivo measure of P-glycoprotein-mediated efflux. In this study we explored the correlation between (99m)Tc-MIBI clearance and histological necrosis following induction chemotherapy and P-glycoprotein expression in osteosarcoma. The primary tumors of 20 patients with high-grade osteosarcoma were imaged at diagnosis with (99m)Tc-MIBI, and the uptake ratios and biological half-lives were calculated. P-Glycoprotein expression in the tumor tissue was determined immunohistochemically and by measuring mRNA expression of the multidrug resistance-1 gene. The histological necrosis following induction chemotherapy was assessed by the Huvos grading system. The biological half-life of (99m)Tc-MIBI ranged from 1.4 to 52.5 h. Seven of the 20 tumor samples had a favorable extent of necrosis following induction chemotherapy. The (99m)Tc-MIBI half-life and uptake ratio showed no correlation with histological necrosis following induction chemotherapy. The (99m)Tc-MIBI half-life and uptake ratio did not correlate with either measure of P-glycoprotein expression. The results of this pilot study indicate that (99m)Tc-MIBI imaging is not an effective predictor of histological necrosis following induction chemotherapy in high-grade osteosarcoma. (99m)Tc-MIBI imaging did not correlate with measures of P-glycoprotein expression in the tumor tissue.


A new algorithm for 3D reconstruction from support functions.
We introduce a new algorithm for reconstructing an unknown shape from a finite number of noisy measurements of its support function. The algorithm, based on a least squares procedure, is very easy to program in standard software such as Matlab, and it works for both 2D and 3D reconstructions (in fact, in principle, in any dimension). Reconstructions may be obtained without any pre- or post-processing steps and with no restriction on the sets of measurement directions except their number, a limitation dictated only by computing time. An algorithm due to Prince and Willsky was implemented earlier for 2D reconstructions, and we compare the performance of their algorithm and ours. But our algorithm is the first that works for 3D reconstructions with the freedom stated in the previous paragraph. Moreover, under mild conditions, theory guarantees that outputs of the new algorithm will converge to the input shape as the number of measurements increases. In addition we offer a linear program version of the new algorithm that is much faster and better, or at least comparable, in performance at low levels of noise and reasonably small numbers of measurements. Another modification of the algorithm, suitable for use in a "focus of attention" scheme, is also described.


Fructose dehydration to 5-hydroxymethylfurfural over solid acid catalysts in a biphasic system.
Different acidic heterogeneous catalysts like alumina, aluminosilicate, zirconium phosphate, niobic acid, ion-exchange resin Amberlyst-15, and zeolite MOR have been studied in fructose dehydration to 5-hydroxymethylfurfural (HMF). The acidity of these materials was characterized using temperature-programmed desorption of NH₃ and IR spectroscopy of adsorbed pyridine. The nature and strength of acid sites was shown to play a crucial role in the selectivity towards HMF. Brønsted acid sites in the case of zeolites and ion-exchange resin led to high selectivities in the dehydration of fructose with an increase in selectivity with the addition of an organic phase. Lewis acidity in the case of phosphate and oxides resulted in the intensive production of humins from fructose at the initial stages of the process, whereas organic phase addition did not affect selectivity.


Venous thromboembolism in critically ill COVID-19 patients receiving prophylactic or therapeutic anticoagulation: a systematic review and meta-analysis.
Many aspects of care such as management of hypercoagulable state in COVID-19 patients, especially those admitted to intensive care units is challenging in the rapidly evolving pandemic of novel coronavirus disease 2019 (COVID-19). We seek to systematically review the available evidence regarding the anticoagulation approach to prevent venous thromboembolism (VTE) among COVID-19 patients admitted to intensive care units. Electronic databases were searched for studies reporting venous thromboembolic events in patients admitted to the intensive care unit receiving any type of anticoagulation (prophylactic or therapeutic). The pooled prevalence (and 95% confidence interval [CI]) of VTE among patients receiving anticoagulant were calculated using the random-effects model. Subgroup pooled analyses were performed with studies reported prophylactic anticoagulation alone and with studies reported mixed prophylactic and therapeutic anticoagulation. We included twelve studies (8 Europe; 2 UK; 1 each from the US and China) in our systematic review and meta-analysis. All studies utilized LMWH or unfractionated heparin as their pharmacologic thromboprophylaxis, either prophylactic doses or therapeutic doses. Seven studies reported on the proportion of patients with the previous history of VTE (range 0-10%). The pooled prevalence of VTE among ICU patients receiving prophylactic or therapeutic anticoagulation across all studies was 31% (95% CI 20-43%). Subgroup pooled analysis limited to studies reported prophylactic anticoagulation alone and mixed (therapeutic and prophylactic anticoagulation) reported pooled prevalences of VTE of 38% (95% CI 10-70%) and 27% (95% CI 17-40%) respectively. With a high prevalence of thromboprophylaxis failure among COVID-19 patients admitted to intensive care units, individualised rather than protocolised VTE thromboprophylaxis would appear prudent at interim.


Iron deficiency in non-anemic chronic obstructive pulmonary disease in a predominantly male population: an ignored entity.
Non-anemic iron deficiency has been studied in heart failure, but studies are lacking in chronic obstructive pulmonary disease (COPD). The potential clinical implications of association of iron deficiency with the severity of COPD warrant research in this direction. This was an observational, cross-sectional study on patients with COPD to compare disease severity, functional status and quality of life in non-anemic patients with COPD between two groups - iron deficient and non-iron deficient. Stable non-anemic COPD with no cause of bleeding were evaluated for serum iron levels, ferritin levels, TIBC, 6MWD, SGRQ, spirometry, and CAT questionnaire. The study patients were divided into iron replete (IR) and iron deficient (ID) groups. A total of 79 patients were studied, out of which 72 were men and seven were women. The mean age was 61.5±8.42 years. Of these, 36 (45.5%; 95% CI, 34.3-56.8%) had iron deficiency. Mean 6-minute-walk distance was significantly shorter in ID (354.28±82.4 meters vs 432.5±47.21 meters; p=0.001). A number of exacerbations in a year were more in ID group (p=0.003), and more patients in ID had at least two exacerbations of COPD within a year (p=0.001). However, the resting pO2, SaO2, and SpO2 levels did not differ significantly between the two groups (p=0.15 and p=0.52, respectively). Also, there was no significant difference in the distribution of patients of a different class of airflow limitations between the two groups. Non-anemic iron deficiency (NAID) is an ignored, yet easily correctable comorbidity in COPD. Patients with iron deficiency have a more severe grade of COPD, had lesser exercise capacity and more exacerbations in a year as compared to non-iron deficient patients. So, foraying into the avenue of iron supplementation, which has shown promising results in improving functional capacity in heart failure and pulmonary hypertension, may well lead to revolutionary changes in the treatment of COPD.


Solution self-assembly of hybrid block copolymers containing poly(ethylene glycol) and amphiphilic beta-strand peptide sequences.
The self-assembly in aqueous solution of hybrid block copolymers consisting of amphiphilic beta-strand peptide sequences flanked by one or two PEG chains was investigated by means of circular dichroism spectroscopy, small-angle X-ray scattering, and transmission electron microscopy. In comparison with the native peptide sequence, it was found that the peptide secondary structure was stabilized against pH variation in the di- and tri-block copolymers with PEG. Small-angle X-ray scattering indicated the presence of fibrillar structures, the dimensions of which are comparable to the estimated width of a beta-strand (with terminal PEG chains in the case of the copolymers). Transmission electron microscopy on selectively stained and dried specimens shows directly the presence of fibrils. It is proposed that these fibrils result from the hierarchical self-assembly of peptide beta-strands into helical tapes, which then stack into fibrils.


Cylindrospermopsin occurrence in two German lakes and preliminary assessment of toxicity and toxin production of Cylindrospermopsis raciborskii (Cyanobacteria) isolates.
Cylindrospermopsis raciborskii, a freshwater cyanobacterium of tropical origin, is not only increasingly found in (sub) tropical water bodies, but also in temperate regions. Since this species may produce potent toxins such as cylindrospermopsin (CYN) and paralytic shellfish poisons, its massive occurrence in water bodies used as drinking water sources or for recreation is of major concern. The proliferation of C. raciborskii in German water bodies has been documented for the past decade. We investigated the occurrence of CYN in field populations and isolates of C. raciborskii from two lakes, and assessed the toxicity of culture isolates using the mouse bioassay, primary rat hepatocytes and human derived cell lines. We show for the first time the occurrence of CYN in German water bodies. None of seven isolates of C. raciborskii contained CYN, however, all isolates were toxic to primary rat hepatocytes, human hepatoblastoma (HEP-G2) and human colon adenocarcinoma (CACO-2) cells. Methanolic extracts were more toxic than aqueous extracts. Three isolates tested in the mouse bioassay were toxic at a concentration of 800 mg kg(-1) showing liver and spleen damage and inflammation of the intestine. These results give strong evidence that the German isolates of C. raciborskii contain currently not identified or unknown toxins.


Adjustment of the ratio between collagenase class II and I improves islet isolation outcome.
Previous studies have clarified the distinct roles of collagenase class I (ccI) and class II (ccII) in enzymatic release of islets from pancreatic tissue. The present study sought to enhance the limited knowledge about the optimal ratio between collagenase classes. Rat islets were isolated utilizing 0.4 DMC-U of neutral protease and 20 PZ-U of fractionated NB-1 collagenase recombined to obtain a ccII/I ratio of 0.5, 1.0, and 1.5. Quality control included assessment of yield (islet equivalents), trypan-blue exclusion, insulin release during static glucose incubation, and transplant function in diabetic nude mice. Data are expressed as mean values +/- SEM. Digestion time was only minimally influenced by different ccII/I ratios. The highest islet yield (P < .05) was obtained using a ccII/I ratio of 1.0. Purity and glucose stimulation index were only marginally affected by different ccII/I ratios. A significant loss of islet viability after 24-hour culture (P < .05) was observed only in islets isolated by means of a ccII/I ratio of 0.5 and 1.5 but not 1.0. Transplantation into diabetic nude mice revealed sustained islet graft function in all experimental groups. The present study indicates that the ratio between ccII and ccI is of significant relevance for optimizing islet yield and viability.


Culture of common carp (Cyprinus carpio) with defined flesh quality for prevention of cardiovascular diseases using finishing feeding strategy.
Fish is the major source of n-3 polyunsaturated fatty acids (n-3 PUFA) which are well known to have positive effects in prevention of cardiovascular diseases. This study investigated the possibility to produce common carp with defined flesh quality using finishing feeding strategy and predict changes of fillet FA by a dilution model. During the 110-day experiment, fish were fed diets with two different vegetable oils (rapeseed/linseed blend, VO; olive oil, OO) only, or with a subsequent fish oil (FO) finishing treatment for 30 or 60 days. Fillet FA composition was measured and data were compared to the ones predicted by the dilution model. The FO finishing treatment resulted in the higher percentage of SFA (from 19.1% to 23.6%; p<0.001), MUFA (from 46.8% to 51.9%; p<0.001), n-3 PUFA (from 3.6% to 7.4%; p<0.001) and lower n-6 PUFA (from 30.5% to 16.9%; p<0.001) and n-6/n-3 ratio (from 8.7 to 2.3; p<0.001) in groups previously fed the VO diet and in lower MUFA percentage (from 67% to 63%; p<0.001) and n-6/n-3 ratio (from 8.2 to 2.8; p<0.001) and higher n-3 PUFA percentage (from 1.5% to 4.5%; p<0.001) in group previously fed the OO diet. The dilution model gave a good prediction for fillet FA changes (slope of the regression line 0.97-1.00; R2 value of 0.992-0.996). The finishing feeding strategy is suggested for production of common carp with a required flesh FA composition for purposes of special nutritional needs, especially for primary and secondary prevention of cardiovascular disease.


Expected gains in life expectancy from various coronary heart disease risk factor modifications.
Despite much evidence that modifying risk factors for coronary heart disease can decrease morbidity and mortality, little is known about the impact of risk-factor modification on life expectancy. We used the Coronary Heart Disease Policy Model, a state-transition computer simulation of the US population, to forecast potential gains in life expectancy from risk-factor modification for the cohort of Americans turning age 35 in 1990. Among 35-year-old men, we projected that the population-wide increase in life expectancy would be about 1.1 years from strict blood pressure control, 0.8 years from smoking cessation, 0.7 years from reduction of serum cholesterol to 200 mg/dl, and about 0.6 years from weight loss to ideal body weight. For women, reducing cholesterol to 200 mg/dl would have the greatest estimated impact-a gain of 0.8 years-whereas smoking cessation, blood pressure control, or weight loss would yield population-wide gains of 0.7, 0.4, and 0.4 years, respectively. Gains for 35-year-old individuals having a given risk factor are greater. We estimate that, on average, male smokers would gain 2.3 years from quitting smoking; males with hypertension would gain 1.1-5.3 years from reducing their diastolic blood pressure to 88 mm Hg; men with serum cholesterol levels exceeding 200 mg/dl would gain 0.5-4.2 years from lowering their serum cholesterol level to 200 mg/dl; and overweight men would gain an average of 0.7-1.7 years from achieving ideal body weight. Corresponding projected gains for at-risk women are 2.8 years from quitting smoking, 0.9-5.7 years from lowering blood pressure, 0.4-6.3 years from decreasing serum cholesterol, and 0.5-1.1 years from losing weight. Eliminating coronary heart disease mortality is estimated to extend the average life expectancy of a 35-year-old man by 3.1 years and a 35-year-old woman by 3.3 years. Population-wide gains in life expectancy from single risk-factor modifications are modest, but gains to individuals at risk can be more substantial.


The gait profile score and movement analysis profile.
The Gait Deviation Index (GDI) has been proposed as an index of overall gait pathology. This study proposes an interpretation of the difference measure upon which the GDI is based, which naturally leads to the definition of a similar index, the Gait Profile Score (GPS). The GPS can be calculated independently of the feature analysis upon which the GDI is based. Understanding what the underlying difference measure represents also suggests that reporting a raw score, as the GPS does, may have advantages over the logarithmic transformation and z-scaling incorporated in the GDI. It also leads to the concept of a Movement Analysis Profile (MAP) to summarise much of the information contained within kinematic data. A validation study on all children attending a paediatric gait analysis service over 3 years (407 children) provides evidence to support the use of the GPS through analysis of its frequency distribution across different Gross Motor Function Classification System (GMFCS) and Gillette Functional Assessment Questionnaire (FAQ) categories, investigation of intra-session variability, and correlation with the square root of GGI. Correlation with GDI confirms the strong relationship between the two measures. The study concludes that GDI and GPS are alternative and closely related measures. The GDI has prior art and is particularly useful in applications arising out of feature analysis such as cluster analysis or subject matching. The GPS will be easier to calculate for new models where a large reference dataset is not available and in association with applications using the MAP.


Vagal nerve stimulation for refractory epilepsy: the surgical procedure and complications in 100 implantations by a single medical center.
In 1997, the US Food and Drug Administration approved the use of intermittent stimulation of the left vagal nerve as adjunctive therapy for seizure control. Vagal nerve stimulation (VNS) has since been considered a safe and effective treatment for medically intractable seizures. The objective of this study is to present our experience with the surgical procedure and outcomes after VNS insertion in the first 100 consecutive patients treated at the Tel-Aviv "Sourasky" Medical Center (TASMC). All patients who underwent VNS device implantation by the authors at TASMC between 2005 and 2011 were studied. The collected data included age at onset of epilepsy, seizure type, duration of epilepsy, age at VNS device implantation, seizure reduction, surgical complications, and adverse effects of VNS over time. Fifty-three males and 47 females, age 21.2 ± 11.1 years, underwent VNS implantation. Indications for surgery were medically refractory epilepsy. The most common seizure type was focal (55 patients, 55 %). Seizure duration until implantation was 14.4 ± 9 years. Mean follow-up time after device insertion was 24.5 ± 22 months. Complications were encountered in 12 patients. The most common complication was local infection (6 patients, 6 %). Six devices were removed-four due to infection and two due to loss of clinical effect. Currently, 63 patients remain in active long-term follow-up; of these, 35 patients have >50 % reduction in frequency of attacks.VNS is a well-tolerated and effective therapeutic alternative in the management of medically refractory epilepsy. The surgical procedure is safe and has a low complication rate.


Targeting chemotherapy to advanced bladder cancer patients most likely to benefit.
Evaluation of: Vickers AJ, Cronin AM, Kattan MW et al.; The International Bladder Cancer Nomogram Consortium: Clinical benefits of a multivariate prediction model for bladder cancer: a decision analytic approach. Cancer 115(23), 5460-5469 (2009). The prognosis from muscle-invasive bladder cancer is poor. Improvements in survival can be made with the use of chemotherapy. The best results are obtained using multiagent regimens, which increase cure rates by approximately 5%. Thus, few patients benefit when compared with treatment morbidity. This low complete response rate makes powering of clinical trials difficult and may prevent them determining which patients benefit most from chemotherapy. Here, we discuss work by Vickers et al. reporting a decision-based analysis using a nomogram to determine the benefit for individual patients from chemotherapy. This decision aid can reduce the number of patients treated by 0.006, without compromising recurrence. The authors conclude that a nomogram-derived 25% risk threshold produced better targeting of chemotherapy than the current standard criteria (mostly using pathological stage).


Estimating usual intakes mainly affects the micronutrient distribution among infants, toddlers and pre-schoolers from the 2012 Mexican National Health and Nutrition Survey.
To compare estimates from one day with usual intake estimates to evaluate how the adjustment for within-person variability affected nutrient intake and adequacy in Mexican children. In order to obtain usual nutrient intakes, the National Cancer Institute's method was used to correct the first 24 h dietary recall collected in the entire sample (n 2045) with a second 24 h recall collected in a sub-sample (n 178). We computed estimates of one-day and usual intakes of total energy, fat, Fe, Zn and Na. 2012 Mexican National Health and Nutrition Survey. A total of 2045 children were included: 0-5·9 months old (n 182), 6-11·9 months old (n 228), 12-23·9 months old (n 537) and 24-47·9 months old (n 1098). From these, 178 provided an additional dietary recall. Although we found small or no differences in energy intake (kJ/d and kcal/d) between one-day v. usual intake means, the prevalence of inadequate and excessive energy intake decreased somewhat when using measures of usual intake relative to one day. Mean fat intake (g/d) was not different between one-day and usual intake among children >6 months old, but the prevalence of inadequate and excessive fat intake was overestimated among toddlers and pre-schoolers when using one-day intake (P6 months. There was overall low variability in energy and fat intakes but higher for micronutrients. Because the usual intake distributions are narrower, the prevalence of inadequate/excessive intakes may be biased when estimating nutrient adequacy if one day of data is used.


Long-term usability and bio-integration of polyimide-based intra-neural stimulating electrodes.
Stimulation of peripheral nerves has transiently restored lost sensation and has the potential to alleviate motor deficits. However, incomplete characterization of the long-term usability and bio-integration of intra-neural implants has restricted their use for clinical applications. Here, we conducted a longitudinal assessment of the selectivity, stability, functionality, and biocompatibility of polyimide-based intra-neural implants that were inserted in the sciatic nerve of twenty-three healthy adult rats for up to six months. We found that the stimulation threshold and impedance of the electrodes increased moderately during the first four weeks after implantation, and then remained stable over the following five months. The time course of these adaptations correlated with the progressive development of a fibrotic capsule around the implants. The selectivity of the electrodes enabled the preferential recruitment of extensor and flexor muscles of the ankle. Despite the foreign body reaction, this selectivity remained stable over time. These functional properties supported the development of control algorithms that modulated the forces produced by ankle extensor and flexor muscles with high precision. The comprehensive characterization of the implant encapsulation revealed hyper-cellularity, increased microvascular density, Wallerian degeneration, and infiltration of macrophages within the endoneurial space early after implantation. Over time, the amount of macrophages markedly decreased, and a layer of multinucleated giant cells surrounded by a capsule of fibrotic tissue developed around the implant, causing an enlargement of the diameter of the nerve. However, the density of nerve fibers above and below the inserted implant remained unaffected. Upon removal of the implant, we did not detect alteration of skilled leg movements and only observed mild tissue reaction. Our study characterized the interplay between the development of foreign body responses and changes in the electrical properties of actively used intra-neural electrodes, highlighting functional stability of polyimide-based implants over more than six months. These results are essential for refining and validating these implants and open a realistic pathway for long-term clinical applications in humans.


Bayesian networks to identify potential high-risk multimorbidity and intervention clusters in inpatients: an explorative data mining study.
Based on large sets of routine hospital data from inpatient cases, we aimed to explore multimorbidity and intervention clusters showing high risks for in-hospital mortality and unplanned readmissions using data-driven analytical methods. We performed an explorative, historical cohort study of consecutive inpatient cases at a tertiary care centre with an integrated platform for routine healthcare data in Switzerland. From January 2012 through to December 2017, all inpatients aged ≥18 years at hospital admission were eligible for study inclusion. We predefined all-cause in-hospital death and unplanned hospital readmission as co-primary outcomes. In a first step, we explored and visualised multimorbidity and intervention clusters using mutual information analysis. In a subsequent step, we trained multi-layer Bayesian networks to identify clusters associated with in-hospital death and/or unplanned hospital readmission. Among 190,837 inpatient cases, 7994 unique diagnoses and 6639 interventions were routinely recorded during the six-year study period. Based on the mutual information analysis, we identified 32 multimorbidity clusters and 24 intervention clusters – of which several were directly related to in-hospital mortality and/or unplanned readmission in the subsequent Bayesian network analysis. Bayesian network analysis may be used as a tool to mine large healthcare databases in order to explore intervention targets for quality improvement programmes. However, the resulting associations should be substantiated in consecutive investigations using specific causal models. (Trial registration no EKNZ 2016-02128.).


Effects of oleuropein on pentylenetetrazol-induced seizures in mice: involvement of opioidergic and nitrergic systems.
Oleuropein, a well-known olive polyphenol, has been shown to mediate neuroprotection in Alzheimer's disease and cerebral ischemia. We investigated the effects of oleuropein on pentylenetetrazole (PTZ)-induced seizures in male NMRI mice, with diazepam as the standard drug. We also examined the possible involvement of opioidergic/nitrergic pathways in the probable effects of oleuropein. Intraperitoneal (i.p.) administration of different doses of oleuropein (10, 20 and 30 mg/kg) significantly increased the seizure threshold 60 min prior to induction of seizure, in a dose-dependent manner. Administration of naltrexone (10 mg/kg, i.p.), an opioid receptor antagonist, completely reversed the anticonvulsant effects of oleuropein (10 mg/kg). On the other hand, the anticonvulsant effect of oleuropein (10 mg/kg) was blocked by a non-effective dose of nonspecific inhibitor of nitric oxide synthase (NOS), L-NAME (1 and 10 mg/kg, i.p) and a selective inhibitor of neuronal NOS, 7-nitroindazole (30 mg/kg, i.p.). However, the nitric oxide precursor, L-arginine (30 and 60 mg/kg, i.p.) potentiated the anticonvulsant activity of oleuropein (10 mg/kg). A selective inducible NOS inhibitor, aminoguanidine (100 mg/kg, i.p.) did not change the anticonvulsant activity of oleuropein. It seems that the opioidergic system and constitutive neuronal NOS may be involved in the anticonvulsant properties of oleuropein.


Processes for managing pathogens.
Wastewater contains human, animal, and plant pathogens capable of causing viral, bacterial, or parasitic infections. There are several routes whereby sewage pathogens may affect human health, including direct contact, contamination of food crops, zoonoses, and vectors. The range and numbers of pathogens in municipal wastewater vary with the level of endemic disease in the community, discharges from commercial activities, and seasonal factors. Regulations to control pathogen risk in the United States and Europe arising from land application of biosolids are based on the concept of multiple barriers to the prevention of transmission. The barriers are (i) treatment to reduce pathogen content and vector attraction, (ii) restrictions on crops grown on land to which biosolids have been applied, and (iii) minimum intervals following application and grazing or harvesting. Wastewater treatment reduces number of pathogens in the wastewater by concentrating them with the solids in the sludge. Although some treatment processes are designed specifically to inactivate pathogens, many are not, and the actual mechanisms of microbial inactivation are not fully understood for all processes. Vector attraction is reduced by stabilization (reduction of readily biodegradable material) and/or incorporation immediately following application. Concerns about health risks have renewed interest in the effects of treatment (on pathogens) and advanced treatment methods, and work performed in the United States suggests that Class A pathogen reduction can be achieved less expensively than previously thought. Effective pathogen risk management requires control to the complete chain of sludge treatment, biosolids handling and application, and post-application activities. This may be achieved by adherence to quality management systems based on hazard analysis critical control point (HACCP) principles.


Effect of smoking on cough reflex sensitivity: basic and preclinical studies.
In healthy nonsmokers, inhalation of one single puff of cigarette smoke immediately evoked airway irritation and cough, which were either prevented or markedly diminished after premedication with hexamethonium. Single-fiber recording experiments performed in anesthetized animals showed that both C fibers and rapidly adapting receptors in the lungs and airways were stimulated by inhalation of one breath of cigarette smoke. Application of nicotine evoked an inward current and triggered depolarization and action potentials in a concentration-dependent manner in a subset of isolated vagal pulmonary sensory neurons. Taken together, these studies showed that activation of the nicotinic acetylcholine receptors expressed on airway sensory nerves is mainly responsible for the acute airway irritation and cough reflex elicited by inhaled cigarette smoke. Chronic exposure to cigarette smoke consistently induces enhanced cough responses to various inhaled tussive agents in guinea pigs. The increased cough sensitivity involves primarily an elevated sensitivity of cough sensors and also an enhanced synaptic transmission of their afferent signals at the nucleus tractus solitaries. In contrast to the observations in animal studies, both enhanced and diminished cough sensitivities to tussive agents have been reported in chronic smokers. This discrepancy is probably related to the history of chronic smoking of the individual smokers and the severity of existing airway inflammation and dysfunction. Furthermore, several other factors possibly contributing to the regulation of cough receptor sensitivity in chronic smokers should also be considered.


An additional electrostatic interaction between adrenodoxin and P450c27 (CYP27A1) results in tighter binding than between adrenodoxin and p450scc (CYP11A1).
Mitochondrial cytochrome P450c27 (product of the CYP27A1 gene) is found to have significantly higher affinity for the common redox partner adrenodoxin than another mitochondrial P450, P450scc (product of the CYP11A1 gene). To investigate the basis of the approximately 30-fold difference in adrenodoxin binding, two sets of P450c27 mutants were generated, expressed in Escherichia coli, and purified. Mutations of one set were within the putative adrenodoxin-binding site containing conserved lysine residues also crucial in P450scc for binding adrenodoxin. The second set included mutations within a sequence aligning with the "meander region" of P450BM-3 proposed to be a site of redox-partner interactions in P450s (Hasemann, C. A., Kurumbail, R. G., Boddupalli, S. S., Peterson, J. A., and Deisenhofer, J. (1995) Structure 3, 41-62). Mutation of the P450c27 conserved lysines (K354A and K358A) led to a approximately 20-fold increase in apparent Ks for adrenodoxin, confirming that these two positively charged residues conserved in mitochondrial P450s are important for adrenodoxin binding. Mutation of Arg-418, conserved in the CYP27A1 family, to serine also decreased the affinity for adrenodoxin approximately 20-fold. This residue is predicted to be located in the meander region. A triple K354A/K358A/R418S mutation profoundly reduced adrenodoxin binding. Thus, in contrast to P450scc, where mutation of the two conserved positively charged residues results in virtually complete inhibition of adrenodoxin binding, in P450c27 there are three of such residues (Lys-354, Lys-358, and Arg-418) important for adrenodoxin interaction.


Efficacy and safety of certolizumab pegol without methotrexate co-administration in Japanese patients with active rheumatoid arthritis: the HIKARI randomized, placebo-controlled trial.
This 24-week, placebo-controlled, double-blind, randomized study (NCT00791921) investigated efficacy and safety of certolizumab pegol (CZP) in Japanese rheumatoid arthritis (RA) patients in whom methotrexate (MTX) cannot be administered. A total of 230 patients were randomized to subcutaneous CZP 200 mg (induction dosing: 400 mg at Weeks 0, 2 and 4) or placebo every 2 weeks. ACR20 responses with CZP were rapid and significant versus placebo at Week 1, sustained to Week 12 (67.2% vs. 14.9%) and Week 24 (63.8% vs. 11.4%). Week 24-modified Total Sharp Score (mTSS) change from baseline (CFB) was 0.48 (CZP) versus 2.45 (placebo). CZP treatment was associated with higher Week 12 ACR20 responses versus placebo (with non-MTX disease modifying antirheumatic drugs [DMARDs], 74.2% vs. 20.0%; without [monotherapy], 59.3% vs. 8.2%) and inhibition of radiographic progression at Week 24 (mTSS CFB; with non-MTX DMARDs, 0.24 vs. 1.61; monotherapy, 0.68 vs. 3.65). Incidences of serious adverse events were 11.2% (CZP) and 2.6% (placebo); one CZP patient died of dissecting aortic aneurysm. CZP treatment with and without non-MTX DMARDs in Japanese patients in whom MTX cannot be administered resulted in rapid, sustained reductions in RA signs and symptoms. Notably, CZP monotherapy showed significant inhibition of radiographic progression.


Noninvasive neuron pinning with nanopillar arrays.
Cell migration in a cultured neuronal network presents an obstacle to selectively measuring the activity of the same neuron over a long period of time. Here we report the use of nanopillar arrays to pin the position of neurons in a noninvasive manner. Vertical nanopillars protruding from the surface serve as geometrically better focal adhesion points for cell attachment than a flat surface. The cell body mobility is significantly reduced from 57.8 μm on a flat surface to 3.9 μm on nanopillars over a 5 day period. Yet, neurons growing on nanopillar arrays show a growth pattern that does not differ in any significant way from that seen on a flat substrate. Notably, while the cell bodies of neurons are efficiently anchored by the nanopillars, the axons and dendrites are free to grow and elongate into the surrounding area to develop a neuronal network, which opens up opportunities for long-term study of the same neurons in connected networks.


Niche and host-associated functional signatures of the root surface microbiome.
Plant microbiomes are critical to host adaptation and impact plant productivity and health. Root-associated microbiomes vary by soil and host genotype, but the contribution of these factors to community structure and metabolic potential has not been fully addressed. Here we characterize root microbial communities of two disparate agricultural crops grown in the same natural soil in a controlled and replicated experimental system. Metagenomic (genetic potential) analysis identifies a core set of functional genes associated with root colonization in both plant hosts, and metatranscriptomic (functional expression) analysis revealed that most genes enriched in the root zones are expressed. Root colonization requires multiple functional capabilities, and these capabilities are enriched at the community level. Differences between the root-associated microbial communities from different plants are observed at the genus or species level, and are related to root-zone environmental factors.


The joint dynamics of investor beliefs and trading during the COVID-19 crash.
We analyze how investor expectations about economic growth and stock returns changed during the February-March 2020 stock market crash induced by the COVID-19 pandemic, as well as during the subsequent partial stock market recovery. We surveyed retail investors who are clients of Vanguard at three points in time: 1) on February 11-12, around the all-time stock market high, 2) on March 11-12, after the stock market had collapsed by over 20%, and 3) on April 16-17, after the market had rallied 25% from its lowest point. Following the crash, the average investor turned more pessimistic about the short-run performance of both the stock market and the real economy. Investors also perceived higher probabilities of both further extreme stock market declines and large declines in short-run real economic activity. In contrast, investor expectations about long-run (10-y) economic and stock market outcomes remained largely unchanged, and, if anything, improved. Disagreement among investors about economic and stock market outcomes also increased substantially following the stock market crash, with the disagreement persisting through the partial market recovery. Those respondents who were the most optimistic in February saw the largest decline in expectations and sold the most equity. Those respondents who were the most pessimistic in February largely left their portfolios unchanged during and after the crash.


Gap-junctional coupling between neurons and astrocytes in primary central nervous system cultures.
Gap-junctional communication between neurons and astrocytes dissociated from rat brain was identified in culture by using dye-transfer assays and electrophysiological measurements. Cell types were identified by using antibodies against beta-tubulin III, glial fibrillary acidic protein, and 2',3'-cyclic-nucleotide phosphohydrolase, which are antigenic determinants of neurons, astroglia, and oligodendrocytes, respectively. Dye coupling was examined as a function of time after dissociated embryonic brain cells were plated onto confluent monolayers of postnatal astrocytes by intracellularly injecting the fluorochrome Lucifer yellow. Coupling of neurons to the astrocytic monolayer was most frequent between 48 h and 72 h in culture and declined over the next 4 days. This gradual uncoupling was accompanied by progressive neuronal maturation, as indicated by morphological measurements in camera lucida drawings. Dye spread was abolished reversibly by octanol, an agent that blocks gap junction channels in other systems. Double whole-cell voltage-clamp measurements confirmed the presence of heterocellular electrical coupling in these cocultures. Coupling was also seen between neurons and astrocytes in cocultures of cells dissociated from embryonic cerebral hemispheres but was rarely detectable in cocultures of postnatal brain cells. These data strongly suggest that junctional communication may provide metabolic and electrotonic interconnections between neuronal and astrocytic networks at early stages of neural development and that such interactions are weakened as differentiation progresses.


Sequencing, analysis, and annotation of expressed sequence tags for Camelus dromedarius.
Despite its economical, cultural, and biological importance, there has not been a large scale sequencing project to date for Camelus dromedarius. With the goal of sequencing complete DNA of the organism, we first established and sequenced camel EST libraries, generating 70,272 reads. Following trimming, chimera check, repeat masking, cluster and assembly, we obtained 23,602 putative gene sequences, out of which over 4,500 potentially novel or fast evolving gene sequences do not carry any homology to other available genomes. Functional annotation of sequences with similarities in nucleotide and protein databases has been obtained using Gene Ontology classification. Comparison to available full length cDNA sequences and Open Reading Frame (ORF) analysis of camel sequences that exhibit homology to known genes show more than 80% of the contigs with an ORF>300 bp and approximately 40% hits extending to the start codons of full length cDNAs suggesting successful characterization of camel genes. Similarity analyses are done separately for different organisms including human, mouse, bovine, and rat. Accompanying web portal, CAGBASE (http://camel.kacst.edu.sa/), hosts a relational database containing annotated EST sequences and analysis tools with possibility to add sequences from public domain. We anticipate our results to provide a home base for genomic studies of camel and other comparative studies enabling a starting point for whole genome sequencing of the organism.


Sub-Epidemics Explain Localized High Prevalence of Reduced Susceptibility to Rilpivirine in Treatment-Naive HIV-1-Infected Patients: Subtype and Geographic Compartmentalization of Baseline Resistance Mutations.
The latest nonnucleoside reverse transcriptase inhibitor (NNRTI) rilpivirine (RPV) is indicated for human immunodeficiency virus type-1 (HIV-1) patients initiating antiretroviral treatment, but the extent of genotypic RPV resistance in treatment-naive patients outside clinical trials is poorly defined. This retrospective observational study of clinical data from Belgium and Portugal evaluates genotypic information from HIV-1 drug-naive patients obtained for the purpose of drug resistance testing. Rilpivirine resistance-associated mutations (RPV-RAMs) were defined based on clinical trials, phenotypic studies, and expert-based resistance algorithms. Viral susceptibility to RPV alone and to the single-tablet regimen was estimated using expert-based resistance algorithms. In 4,631 HIV-1 treatment-naive patients infected with diverse HIV-1 subtypes, major RPV-RAMs were detected in 4.6%, while complete viral susceptibility to RPV was estimated in 95% of patients. Subtype C- and F1-infected patients displayed the highest levels of reduced viral susceptibility at baseline, respectively 13.2% and 9.3%, mainly due to subtype- and geographic-dependent occurrence of RPV-RAMs E138A and A98G as natural polymorphisms. Strikingly, a founder effect in Portugal resulted in a 138A prevalence of 13.2% in local subtype C-infected treatment-naive patients. The presence of transmitted drug resistance did not impact our estimates. RPV is the first HIV-1 inhibitor for which, in the absence of transmitted drug resistance, intermediate or high-level genotypic resistance can be detected in treatment-naive patients. The extent of RPV susceptibility in treatment-naive patients differs depending on the HIV-1 subtype and dynamics of local compartmentalized epidemics. The highest prevalence of reduced susceptibility was found to be 15.7% in Portuguese subtype C-infected treatment-naive patients. In this context, even in the absence of transmitted HIV-1 drug resistance (TDR), drug resistance testing at baseline should be considered extremely important before starting treatment with this NNRTI.


A geographic cline induced by negative frequency-dependent selection.
Establishment of geographic morph frequency clines is difficult to explain in organisms with limited gene flow. Balancing selection, such as negative frequency-dependent selection (NFDS), is instead suggested to establish a morph frequency cline on a geographic scale at least theoretically. Here we tested whether a large-scale smooth cline in morph frequency is established by NFDS in the female-dimorphic damselfly, Ischnura senegalensis, where andromorphs and gynomorphs are maintained by NFDS. We found a large-scale latitudinal cline in the morph frequency: andromorph frequency ranged from 0.05 (South) to 0.79 (North). Based on the empirical data on the numbers of eggs, the number of ovariole, abdomen length and latitude, the potential fitness of andromorphs was estimated to be lower than that of gynomorphs in the south, and higher in the north, suggesting the gene-by-environment interaction. From the morph-specific latitudinal cline in potential fitness, the frequency of andromorphs was expected to shift from 0 to 1 without NFDS, because a morph with higher potential fitness wins completely and the two morphs will switch at some point. In contrast, NFDS led to the coexistence of two morphs with different potential fitness in a certain geographic range along latitude due to rare morph advantage, and resulted in a smooth geographic cline of morph frequency. Our results provide suggestive evidence that the combination of NFDS and gene-by-environment interaction, i.e., multi-selection pressure on color morphs, can explain the geographic cline in morph frequency in the current system.


Genetic population structure of the malaria vector Anopheles baimaii in north-east India using mitochondrial DNA.
Anopheles baimaii is a primary vector of human malaria in the forest settings of Southeast Asia including the north-eastern region of India. Here, the genetic population structure and the basic population genetic parameters of An. baimaii in north-east India were estimated using DNA sequences of the mitochondrial cytochrome oxidase sub unit II (COII) gene. Anopheles baimaii were collected from 26 geo-referenced locations across the seven north-east Indian states and the COII gene was sequenced from 176 individuals across these sites. Fifty-seven COII sequences of An. baimaii from six locations in Bangladesh, Myanmar and Thailand from a previous study were added to this dataset. Altogether, 233 sequences were grouped into eight population groups, to facilitate analyses of genetic diversity, population structure and population history. A star-shaped median joining haplotype network, unimodal mismatch distribution and significantly negative neutrality tests indicated population expansion in An. baimaii with the start of expansion estimated to be ~0.243 million years before present (MYBP) in north-east India. The populations of An. baimaii from north-east India had the highest haplotype and nucleotide diversity with all other populations having a subset of this diversity, likely as the result of range expansion from north-east India. The north-east Indian populations were genetically distinct from those in Bangladesh, Myanmar and Thailand, indicating that mountains, such as the Arakan mountain range between north-east India and Myanmar, are a significant barrier to gene flow. Within north-east India, there was no genetic differentiation among populations with the exception of the Central 2 population in the Barail hills area that was significantly differentiated from other populations. The high genetic distinctiveness of the Central 2 population in the Barail hills area of the north-east India should be confirmed and its epidemiological significance further investigated. The lack of genetic population structure in the other north-east Indian populations likely reflects large population sizes of An. baimaii that, historically, were able to disperse through continuous forest habitats in the north-east India. Additional markers and analytical approaches are required to determine if recent deforestation is now preventing ongoing gene flow. Until such information is acquired, An. baimaii in north-east India should be treated as a single unit for the implementation of vector control measures.


Clearance of cervical human papillomavirus infection by topical application of curcumin and curcumin containing polyherbal cream: a phase II randomized controlled study.
Curcumin and curcumin containing polyherbal preparations have demonstrated anti-microbial and anti- viral properties in pre-clinical studies. Till date no therapeutic intervention has been proved to be effective and safe in clearing established cervical human papillomavirus (HPV) infection. The present study evaluated the efficacy of Basant polyherbal vaginal cream (containing extracts of curcumin, reetha, amla and aloe vera) and of curcumin vaginal capsules to eliminate HPV infection from cervix. Women were screened by Pap smear and HPV DNA test by PCR. HPV positive women without high grade cervical neoplasias (N=287) were randomized to four intervention arms to be treated with vaginal Basant cream, vaginal placebo cream, curcumin vaginal capsules and placebo vaginal capsules respectively. All subjects were instructed to use one application of the assigned formulation daily for 30 consecutive days except during menstruation and recalled within seven days of the last application for repeat HPV test, cytology and colposcopy. HPV clearance rate in Basant arm (87.7%) was significantly higher than the combined placebo arms (73.3%). Curcumin caused higher rate of clearance (81.3%) than placebo though the difference was not statistically significant. Vaginal irritation and itching, mostly mild to moderate, was significantly higher after Basant application. No serious adverse events were noted.


Changes in extracellular pH mediate the chronotropic responses to L-arginine.
We have recently shown that exogenous nitric oxide (NO) elicits a positive chronotropic response by stimulating the hyperpolarization activated current, I(f). To examine whether L-arginine (L-Arg) can mimic the chronotropic effect of NO by enhancing its endogenous production. In spontaneously beating guinea pig atria we evaluated the heart rate (HR) response to increasing concentrations of L-Arg (1 mumol/l to 10 mmol/l), and compared it with that for D-Arg or L-lysine (L-Lys) (all in free base (FB) or hydrochloride (HCl) formulation). L-ArgFB > 100 mumol/l caused a reversible dose-dependent increase in HR (peak effect +64 +/- 7 bpm at 10 mmol/l, P < 0.05, n = 8). However, a similar HR response occurred with D-ArgFB (n = 7) or L-LysFB (n = 6). All FB formulations increased the perfusate pH (peak [pH]o = 8.61 +/- 0.03). Although alkalinization can stimulate NO release from the endothelium, this is unlikely to have contributed to HR changes in our preparation, since neither NG-methyl-L-arginine, (100-500 mumol/l, which per se reduced HR by 8 +/- 1%, P < 0.05, n = 9) nor NO scavenging (fresh 5% red blood cells, n = 9) caused a rightward shift of the concentration-response curve to L-ArgFB. Furthermore, as opposed to FB formulations, L-ArgHCl, D-ArgHCl or L-LysHCl > 1 mmol/l significantly decreased HR and [pH]o (n = 17). The chronotropic effects of L-ArFB or L-ArgHCl were reproduced by changing [pH]o with NaOH (n = 8) or HCl (n = 7), whereas the HR increase with L-ArgFB was prevented by clamping [pH]o at 7.42 +/- 0.07 (n = 10). In vitro, L-Arg can markedly affect HR through a pH-mediated, NO-independent mechanism. Our data show that the opposing changes in [pH]o induced by different formulations of L-Arg can importantly confound the assessment of the biological effects of this amino acid.


Resolution testing of coherent optical systems by means of linear gratings.
Testing of coherent optical data processing systems has shown a need for a method which measures resolution at all points in the image format. The use of multifrequency linear gratings as test objects satisfies this need. Quantitative data may be obtained if care is taken to calibrate and linearize the photographic process. However, the method's principal application seems to be for qualitative testing. A comparison is made between calculated wavefront aberrations and experimental results obtained using linear gratings.