Molecular therapy for peritoneal dissemination of xenotransplanted human MKN-45 gastric cancer cells with adenovirus mediated Bax gene transfer.
Gene therapy is an innovative therapeutic approach for cancer. An adenoviral vector expressing the tumour suppressor p53 gene (Ad/p53) is currently under clinical evaluation for various cancers. We recently developed a binary adenoviral vector system that can express the strong proapoptotic gene Bax (Ad/PGK-GV16+Ad/GT-Bax: Ad/Bax). To evaluate the potential of Bax gene therapy for gastric cancer, we assessed its antitumour effect in comparison with that of p53. The human gastric cancer cell lines MKN-1, MKN-7, MKN-28, and MKN-45 were treated with Ad/Bax or Ad/p53, and cell viability, transgene expression, and caspase activation were assessed in vitro. To compare the antitumour effects of Ad/Bax and Ad/p53 treatment in vivo, subcutaneous tumours and peritoneal dissemination of MKN-45 cells were generated in nude mice. Each mouse underwent intratumoral or intraperitoneal administration of viruses and the growth of implanted tumours was observed after treatment. Treatment with Ad/Bax and Ad/p53 resulted in marked Bax and p53 protein expression and effective apoptosis induction in MKN-1, MKN-7, and MKN-28 cells in vitro. In contrast, MKN-45 cells showed resistance to Ad/p53 and only treatment with Ad/Bax resulted in activation of caspase 3 expression and massive apoptosis. Ad/Bax treatment was more effective in suppressing both subcutaneous and peritoneally disseminated MKN-45 tumours compared with Ad/p53 treatment. Ad/Bax treatment significantly inhibited the growth of even p53 resistant gastric cancer in vitro and in vivo. Therefore, adenovirus mediated Bax gene transfer may be useful in gene therapy for gastric cancers.


Protective role of Ginkgo biloba on petroleum wastewater-induced toxicity in Vicia faba L. (Fabaceae) root tip cells.
The Melet is one of Turkey's economically important rivers. Most of the petroleum plants are located at fairly nearby of the river This situation is considered as main source of heavy metal pollution in the river. The present study was designed to evaluate the protective role of Ginkgo biloba (GB) on cytotoxicity induced by petroleum wastewater in Vicia faba root tip cells. For this aim, we used the germination percentage, root length, weight gain and micronucleus (MN) frequency as indicators of cytotoxicity. Additionally to the cytological analysis, lipid peroxidation analyses were also performed in V. faba roots. Heavy metal concentrations in wastewater were measured by atomic absorption spectrophotometer (AAS). The V. faba seeds were divided into six groups. They were treated with petroleum wastewater and 10, 20 and 30 microM doses of GB. As a result, the mean concentrations of heavy metals in wastewater were observed in the order: Pb > A > Ni > Cr > Fe > Cu > Zn > Cd. The highest germination percentage was observed in the seeds of the control and positive control groups (in proportion as 98 and 96%, respectively). Wastewater treatment caused a significant decrease in the germination percentage of Group III (in proportion as 44%). The highest root length and weight gain were observed in the seeds of the control and positive control groups at the end of the experimental period. The least root length and weight gain were observed in the seeds of Group III treated with wastewater alone. In the control group, the final weights of all the seeds increased about 4.08 g according to initial weight. The root lengths of the control seeds were measured as 6.80 cm at the end of the experimental period. The final weights of the seeds exposed to wastewater alone increased about 0.90 g according to initial weight. Besides, there was a significantly increase in the MDA levels of the roots exposed to wastewater. Heavy metals in wastewater significantly affected the MDA production indicating lipid peroxidation. But, GB-treatment caused amelioration in indices of the germination percentage, root length, weight gain, MN frequency and lipid peroxidation when compared with group III. Each dose of GB provided protection against wastewater toxicity and its strongest protective effect observed at dose of 30 microM. In vivo results showed that GB is a potential protector against toxicity induced by petroleum wastewater and its protective role is dose-dependent.


The protocadherin 17 gene affects cognition, personality, amygdala structure and function, synapse development and risk of major mood disorders.
Major mood disorders, which primarily include bipolar disorder and major depressive disorder, are the leading cause of disability worldwide and pose a major challenge in identifying robust risk genes. Here, we present data from independent large-scale clinical data sets (including 29 557 cases and 32 056 controls) revealing brain expressed protocadherin 17 (PCDH17) as a susceptibility gene for major mood disorders. Single-nucleotide polymorphisms (SNPs) spanning the PCDH17 region are significantly associated with major mood disorders; subjects carrying the risk allele showed impaired cognitive abilities, increased vulnerable personality features, decreased amygdala volume and altered amygdala function as compared with non-carriers. The risk allele predicted higher transcriptional levels of PCDH17 mRNA in postmortem brain samples, which is consistent with increased gene expression in patients with bipolar disorder compared with healthy subjects. Further, overexpression of PCDH17 in primary cortical neurons revealed significantly decreased spine density and abnormal dendritic morphology compared with control groups, which again is consistent with the clinical observations of reduced numbers of dendritic spines in the brains of patients with major mood disorders. Given that synaptic spines are dynamic structures which regulate neuronal plasticity and have crucial roles in myriad brain functions, this study reveals a potential underlying biological mechanism of a novel risk gene for major mood disorders involved in synaptic function and related intermediate phenotypes.


Population-based estimates of the occurrence of multiple vs first primary basal cell carcinomas in 4 European regions.
To estimate the population-based incidence of first and multiple basal cell carcinomas (BCCs) throughout Europe. The registry practices of 4 population-based cancer registries that routinely register BCC incidence were evaluated for inclusion of first and subsequent histologically confirmed BCCs. Where multiple BCCs were not routinely registered, comparisons with hospital databases were made. Cancer registry databases from Finland, Malta, the Netherlands (Eindhoven), and Scotland were inspected for registry of first and multiple BCCs in recent years. Cross-checks with hospital and pathology databases were made to check for completeness. Age-standardized first BCC incidence rates varied between 77 (Malta) and 158 (Eindhoven) per 100 000 person-years. Generally, rates were higher in males than in females, and incidences increased steeply with increasing age. There were approximately 30% more patients with a BCC and 40% to 100% more BCC tumors diagnosed in a given calendar year than were routinely reported for patients with a first primary BCC. The difference between the number of first primary BCCs and the total number of BCCs in a calendar year was generally slightly higher for males than for females and increased substantially with increasing age. Currently, routinely reported first BCC incidence rates of the included countries should be multiplied by a factor of 1.3 for an estimate of total number of patients diagnosed as having a BCC in a given year.


Incidence and Risk Factors for Volar Wrist Ganglia in the U.S. Military and Civilian Populations.
To identify the incidence and demographic factors associated with volar wrist ganglia in both military and civilian beneficiary populations. The U.S. Department of Defense Management Analysis and Reporting Tool (M2) accesses a comprehensive database of all health care visits by military personnel and their dependents. Because there is no specific code for ganglions of the wrist, the database was searched for all military personnel and civilian beneficiaries with an International Classification of Diseases, 9th Revision, diagnosis of 727.41 (ganglion of a joint) or 727.43 (ganglion, unspecified location) between 2009 and 2014. Two random samples of 1000 patients were selected from both the military and the civilian beneficiary cohorts, and their electronic medical records were examined to identify those with volar wrist ganglia. The proportion of volar wrist ganglia was then applied to the overall population data to estimate the total incidence with a 95% confidence interval and 5% margin of error. Unadjusted incidence rates and adjusted incidence rate ratios were determined using Poisson regression, controlling for age, sex, branch of military service, and military seniority. The unadjusted incidence of volar wrist ganglia is 3.72 per 10,000 person-years (0.04%/y) in female civilian beneficiaries, 1.04 per 10,000 person-years (0.01%/y) in male civilian beneficiaries, 7.98 per 10,000 person-years (0.08%/y) in female military personnel, and 3.73 per 10,000 person-years (0.04%/y) in male military personnel. When controlled for age, military personnel have a 2.5-times increased rate of volar wrist ganglia, and women have a 2.3-times increased rate. In the military cohort, female sex, branch of service, and seniority were significantly associated with the diagnosis of a volar wrist ganglion when controlled for age. In the civilian beneficiary cohort, only female sex was significant. Military service members have higher rates of volar wrist ganglia diagnoses than their age- and sex-matched civilian counterparts. Women are significantly more likely to be diagnosed with a volar wrist ganglion, regardless of age or military status. The epidemiology of volar wrist ganglia is poorly defined, and few studies have firmly defined demographic factors associated with the diagnosis. We provide the overall incidence rate of the diagnosis and report a significant association with female sex even when controlled for age.


Thujone exhibits low affinity for cannabinoid receptors but fails to evoke cannabimimetic responses.
Absinthe, an abused drug in the early 1900s, has been speculated to activate the receptors responsible for marijuana intoxication (the CB1 cannabinoid receptor) (Nature 253:365-356; 1975). To test this hypothesis, we investigated oil of wormwood (Artemisia absinthium) the active plant product found in absinthe, and thujone, the active compound found in oil of wormwood. Radioligand receptor binding assays employing membrane preparations from rat brains containing CB1 cannabinoid receptors, and human tonsils containing CB2 receptors, demonstrated that thujone displaced [3H]CP55940, a cannabinoid agonist, only at concentrations above 10 microM. HPLC analysis of oil of wormwood revealed that only the fractions having mobility close to thujone displaced [3H]CP55940 from the CB1 cannabinoid receptor. [35S]GTPgammaS binding assays revealed that thujone failed to stimulate G-proteins even at 0.1 mM. Thujone failed to inhibit forskolin-stimulated adenylate cyclase activity in N18TG2 membranes at 1 mM. Rats administered thujone exhibited different behavioral characteristics compared with rats administered a potent cannabinoid agonist, levonantradol. Therefore, the hypothesis that activation of cannabinoid receptors is responsible for the intoxicating effects of thujone is not supported by the present data.


Analysis of code relating sequences to conformation in globular prtoeins. Theory and application of expected information.
1. An information theory analysis of the folding of a globular protein is proposed. 2. The folding is seen as a transfer of information between two messages, the primary sequence and the biologically active conformation. 3. It is shown how the information transferred was estimated by inspection of proteins of known primary sequence and conformation. 4. In this estimation, concerted use of subjective (Bayesian) probabilities leads to a more robust approach which can be employed whether the number of proteins of known sequence and conformation is large or small. 5. Further, it is demonstrated that the problem then becomes a very simple algebraic formulation for information estimates. 6. Finally, it is shown how this process of information theory analysis can be reversed to predict the conformation of a protein by using its primary sequence and the above information estimates obtained from other proteins. 7. The present paper provides the theoretical basis for the derivation and application of a stereochemical alphabet (Robson & Pain, 1974a,c), and for an investigation of the effects of residues on the conformations of their neighbours (Robson & Pain, 1974b).


How to make stripes: deciphering the transition from non-periodic to periodic patterns in Drosophila segmentation.
The generation of metameric body plans is a key process in development. In Drosophila segmentation, periodicity is established rapidly through the complex transcriptional regulation of the pair-rule genes. The 'primary' pair-rule genes generate their 7-stripe expression through stripe-specific cis-regulatory elements controlled by the preceding non-periodic maternal and gap gene patterns, whereas 'secondary' pair-rule genes are thought to rely on 7-stripe elements that read off the already periodic primary pair-rule patterns. Using a combination of computational and experimental approaches, we have conducted a comprehensive systems-level examination of the regulatory architecture underlying pair-rule stripe formation. We find that runt (run), fushi tarazu (ftz) and odd skipped (odd) establish most of their pattern through stripe-specific elements, arguing for a reclassification of ftz and odd as primary pair-rule genes. In the case of run, we observe long-range cis-regulation across multiple intervening genes. The 7-stripe elements of run, ftz and odd are active concurrently with the stripe-specific elements, indicating that maternal/gap-mediated control and pair-rule gene cross-regulation are closely integrated. Stripe-specific elements fall into three distinct classes based on their principal repressive gap factor input; stripe positions along the gap gradients correlate with the strength of predicted input. The prevalence of cis-elements that generate two stripes and their genomic organization suggest that single-stripe elements arose by splitting and subfunctionalization of ancestral dual-stripe elements. Overall, our study provides a greatly improved understanding of how periodic patterns are established in the Drosophila embryo.


Development of an intervention for moral injury-related mental health difficulties in UK military veterans: a feasibility pilot study protocol.
Background: Experiencing potentially morally injurious events (PMIEs) has been found to be significantly associated with poor mental health outcomes in military personnel/veterans. Currently, no manualised treatment for moral injury-related mental health difficulties for UK veterans exists. This article describes the design, methods and expected data collection of the Restore & Rebuild (R&R) protocol, which aims to develop procedures to treat moral injury related mental ill health informed by a codesign approach. Methods: The study consists of three main stages. First, a systematic review will be conducted to understand the best treatments for the symptoms central to moral injury-related mental ill health (stage 1). Then the R&R manual will be co-designed with the support of UK veteran participants with lived experience of PMIEs as well as key stakeholders who have experience of supporting moral injury affected individuals (stage 2). The final stage of this study is to conduct a pilot study to explore the feasibility and acceptability of the R&R manual (stage 3). Results: Qualitative data will be analysed using thematic analysis. Conclusions: This study was approved by the King's College London's Research Ethics Committee (HR-20/21-20850). The findings will be disseminated in several ways, including publication in academic journals, a free training event and presentation at conferences. By providing information on veteran, stakeholder and clinician experiences, we anticipate that the findings will not only inform the development of an acceptable evidence-based approach for treating moral injury-related mental health problems, but they may also help to inform broader approaches to providing care to trauma exposed military veterans.


Mechanisms of orexin-induced depolarizations in rat dorsal motor nucleus of vagus neurones in vitro.
1. Whole-cell patch-clamp recordings were made from neurones of the dorsal motor nucleus of the vagus (DMNV), including Fluoro-gold-labelled parasympathetic preganglionic neurones (PPNs), in slices of the rat medulla. In the latter case, rats had received an I.P. injection of Fluoro-gold solution (10 microg) 2-3 days earlier. 2. Superfusion of orexin A or B (10-300 nM) caused a slow depolarization in approximately 30% of the DMNV neurones, including PPNs. Orexin-induced depolarizations, which persisted in TTX (0.5 microM)-containing Krebs solution, were reduced by 70% in a low-Na+ (26 mM) Krebs solution, indicating the involvement of Na+ ions. A significant change in orexin-induced depolarizations was not obtained in either a high-K+ (7 mM) or Cd2+ (100 microM) Krebs solution. 3. Inclusion of the hydrolysis-resistant guanine nucleotide GDP-beta-S in the patch solution significantly reduced the orexin A- or B-induced depolarizations. 4. Under whole-cell voltage-clamp conditions, the orexin-induced inward current declined with hyperpolarization, but did not reverse polarity in the potential range between -120 and 0 mV. In low-Na+ solution, the orexin-induced current was reduced, and the I-V curve reversed polarity at about -105 mV; the response was further reduced and the reversal potential shifted to -90 mV in a low-Na+, high-K+ Krebs solution. 5. It is concluded that the peptides orexin A and B, acting on orexin receptors, which are GTP-binding-protein coupled, are excitatory to DMNV neurones. In addition, more than one conductance, which may include a non-selective cation conductance and a K+ conductance, appears to be involved in the orexin-induced depolarization.


Exploiting vulnerabilities of cancer by targeting nuclear receptors of stromal cells in tumor microenvironment.
The tumor microenvironment is a complex and dynamic cellular community comprising the tumor epithelium and various tumor-supporting cells such as immune cells, fibroblasts, immunosuppressive cells, adipose cells, endothelial cells, and pericytes. The interplay between the tumor microenvironment and tumor cells represents a key contributor to immune evasiveness, physiological hardiness and the local and systemic invasiveness of malignant cells. Nuclear receptors are master regulators of physiological processes and are known to play pro-/anti-oncogenic activities in tumor cells. However, the actions of nuclear receptors in tumor-supporting cells have not been widely studied. Given the excellent druggability and extensive regulatory effects of nuclear receptors, understanding their biological functionality in the tumor microenvironment is of utmost importance. Therefore, the present review aims to summarize recent evidence about the roles of nuclear receptors in tumor-supporting cells and their implications for malignant processes such as tumor proliferation, evasion of immune surveillance, angiogenesis, chemotherapeutic resistance, and metastasis. Based on findings derived mostly from cell culture studies and a few in vivo animal cancer models, the functions of VDR, PPARs, AR, ER and GR in tumor-supporting cells are relatively well-characterized. Evidence for other receptors, such as RARβ, RORγ, and FXR, is limited yet promising. Hence, the nuclear receptor signature in the tumor microenvironment may harbor prognostic value. The clinical prospects of a tumor microenvironment-oriented cancer therapy exploiting the nuclear receptors in different tumor-supporting cells are also encouraging. The major challenge, however, lies in the ability to develop a highly specific drug delivery system to facilitate precision medicine in cancer therapy.


Decreased somatosensory activity to non-threatening touch in combat veterans with posttraumatic stress disorder.
Posttraumatic stress disorder (PTSD) is a severe psychiatric disorder prevalent in combat veterans. Previous neuroimaging studies have demonstrated that patients with PTSD exhibit abnormal responses to non-threatening visual and auditory stimuli, but have not examined somatosensory processing. Thirty male combat veterans, 16 with PTSD and 14 without, completed a tactile stimulation task during a 306-sensor magnetoencephalography (MEG) recording. Significant oscillatory neural responses were imaged using a beamforming approach. Participants also completed clinical assessments of PTSD, combat exposure, and depression. We found that veterans with PTSD exhibited significantly reduced activity during early (0-125 ms) tactile processing compared with combat controls. Specifically, veterans with PTSD had weaker activity in the left postcentral gyrus, left superior parietal area, and right prefrontal cortex in response to nonthreatening tactile stimulation relative to veterans without PTSD. The magnitude of activity in these brain regions was inversely correlated with symptom severity, indicating that those with the most severe PTSD had the most abnormal neural responses. Our findings are consistent with a resource allocation view of perceptual processing in PTSD, which directs attention away from nonthreatening sensory information.


A comparative multi-parametric in vitro model identifies the power of test conditions to predict the fibrotic tendency of a biomaterial.
Despite growing effort to advance materials towards a low fibrotic progression, all implants elicit adverse tissue responses. Pre-clinical biomaterial assessment relies on animals testing, which can be complemented by in vitro tests to address the Russell and Burch's 3R aspect of reducing animal burden. However, a poor correlation between in vitro and in vivo biomaterial assessments confirms a need for suitable in vitro biomaterial tests. The aim of the study was to identify a test setting, which is predictive and might be time- and cost-efficient. We demonstrated how sensitive in vitro biomaterial assessment based on human primary macrophages depends on test conditions. Moreover, possible clinical scenarios such as lipopolysaccharide contamination, contact to autologous blood plasma, and presence of IL-4 in an immune niche influence the outcome of a biomaterial ranking. Nevertheless, by using glass, titanium, polytetrafluorethylene, silicone, and polyethylene representing a specific material-induced fibrotic response and by comparison to literature data, we were able to identify a test condition that provides a high correlation to state-of-the-art in vivo studies. Most important, biomaterial ranking obtained under native plasma test conditions showed a high predictive accuracy compared to in vivo assessments, strengthening a biomimetic three-dimensional in vitro test platform.


All-optical technique for modulation format conversion from NRZ-OOK to RZ-16QAM employing nonlinear optical loop mirror with 1:2 coupler.
All-optical technique for modulation format conversion from 4 channels non-return-to-zero on-off-keying (OOK) to return-to-zero 16 quadrature amplitude modulation (16QAM) employing nonlinear optical loop mirror with 1:2 coupler configuration is proposed and experimentally demonstrated at 10Gs/s. The experimentally converted 16QAM signal was distorted in its phase by cross-phase modulation induced amplitude-to-phase noise conversion. The effect of amplified spontaneous emission noise in the amplified OOK signals to the converted 16QAM's phase was theoretically discussed.


CauloBrowser: A systems biology resource for Caulobacter crescentus.
Caulobacter crescentus is a premier model organism for studying the molecular basis of cellular asymmetry. The Caulobacter community has generated a wealth of high-throughput spatiotemporal databases including data from gene expression profiling experiments (microarrays, RNA-seq, ChIP-seq, ribosome profiling, LC-ms proteomics), gene essentiality studies (Tn-seq), genome wide protein localization studies, and global chromosome methylation analyses (SMRT sequencing). A major challenge involves the integration of these diverse data sets into one comprehensive community resource. To address this need, we have generated CauloBrowser (www.caulobrowser.org), an online resource for Caulobacter studies. This site provides a user-friendly interface for quickly searching genes of interest and downloading genome-wide results. Search results about individual genes are displayed as tables, graphs of time resolved expression profiles, and schematics of protein localization throughout the cell cycle. In addition, the site provides a genome viewer that enables customizable visualization of all published high-throughput genomic data. The depth and diversity of data sets collected by the Caulobacter community makes CauloBrowser a unique and valuable systems biology resource.


Molecular cloning, expression, and chromosomal localization of the human earliest lymphocyte activation antigen AIM/CD69, a new member of the C-type animal lectin superfamily of signal-transmitting receptors.
The activation of T lymphocytes, both in vivo and in vitro, induces the expression of CD69. This molecule, which appears to be the earliest inducible cell surface glycoprotein acquired during lymphoid activation, is involved in lymphocyte proliferation and functions as a signal transmitting receptor in lymphocytes, natural killer (NK) cells, and platelets. To determine the structural basis for CD69 function, the cDNA coding for CD69 was isolated by a polymerase chain reaction-based strategy using oligonucleotides deduced from peptide sequences of the purified protein. The isolated cDNA exhibited a single open reading frame of 597 bp coding for CD69, and predicted a 199-amino acid protein of type II membrane topology, with extracellular (COOH-terminal), transmembrane, and intracellular domains. The CD69 clone hybridized to a 1.7-kb mRNA species, which was rapidly induced and degraded after lymphocyte stimulation, consistent with the presence of rapid degradation signals at the 3' untranslated region. Transient expression of the polypeptide encoded by CD69 cDNA in COS-7 cells demonstrated that it presented properties comparable to native CD69 protein. The CD69 gene was regionally mapped to chromosome 12 p13-p12 by both somatic cell hybrid DNA analysis and fluorescence in situ hybridization coupled with GTG banding (G bands by trypsin using Giemsa). Protein sequence homology search revealed that CD69 is a new member of the Ca(2+)-dependent (C-type) lectin superfamily of type II transmembrane receptors, which includes the human NKG2, the rat NKR-P1, and the mouse NKR-P1 families of NK cell-specific genes. CD69 also has a structural homology with other type II lectin cell surface receptors, such as the T cell antigen Ly49, the low avidity immunoglobulin E receptor (CD23), and the hepatic asialoglycoprotein receptors. The CD69 protein also shares functional characteristics with most members of this superfamily, which act as transmembrane signaling receptors in early phases of cellular activation.


Infectious SIV resides in adipose tissue and induces metabolic defects in chronically infected rhesus macaques.
HIV reservoirs pose major challenges to viral eradication. The main cellular reservoirs include CD4 T cells and macrophages, whereas anatomic reservoirs are thought to be primarily lymphoid tissues. Adipose tissue represents a potentially important non-lymphoid location for HIV replication and persistence because the stromal-vascular-fraction (AT-SVF) contains activated innate and adaptive immune cells that increase in number during infections, obesity, and chronic inflammation. Adipose tissue from two groups of SHIV-SF162p3-infected (~4 weeks acute infection) or SIVmac251-infected (~38 weeks chronic infection) rhesus macaques (N = 8 for each group) were studied for immune cell content, viral infectiousness, and metabolic health. The AT-SVF cells from SHIV-infected monkeys contained abundant memory CD4 and CD8 T cells, with fewer NKT cells and macrophages, and no B cells. Proviral DNA (Gag and Env) was readily detectable by nested PCR in AT-SVF cells from multiple adipose depots (subcutaneous and visceral) of acutely infected monkeys, but mostly from visceral fat. More importantly, viral outgrowth assays using input CD4 T cells derived from AT-SVF cells or peripheral blood of chronically infected monkeys resulted in robust replication of infectious virus from both AT-SVF and peripheral blood CD4 T cells. Chronically infected monkeys also experienced adipocyte dysfunction (suppression of major adipogenic genes) and systemic dyslipidemia (decreased serum total cholesterol and free fatty acids, and increased triglycerides), similar to metabolic abnormalities of HIV patients. Adipose tissues of SIV-infected rhesus macaques become major compartments for infected immune cells, which in turn induce defects in adipose tissue metabolism.


Prediction of anthracycline induced cardiotoxicity: study of thirty-one Iraqi adult patients.
To look for a nearly ideal tool for prediction of anthracycline-induced cardiotoxicity. Thirty-one patients with various hematological malignancies were included in the study which was conducted from Sept. 2005 to Sept. 2006 in Baghdad Teaching Hospital - Hematology Unit. Initial cardiovascular assessment including cardiac troponin I, electrocardiography and echocardiography were done and repeated one month after the commencement of anthracycline-based regimen. Cardiotoxicity was considered present if the patient has clinical and electrocardiographic evidences, troponin positivity, echocardiographic evidence, or any combination of these. The mean age for the study sample was 34.1 ± 17.2 years comprising of 17 male and 14 female patients. Increasing age, body surface area, anthracycline dose as well as the concomitant use of cyclophosphomide/All Trans Retinoic Acid were associated with increased risk of cardiotoxicity. The cut-off point of body surface area above which the risk of anthracycline-induced cardiotoxicity is increased was 1.88 m2 while the cut-off point for anthracyclines dose was 145.5 mg/m2. The constellation of clinical data, ECG, and cTnI was 92% predictive of early evidence of anthracycline-induced cardiotoxicity. More weight is added when echocardiography is used as a diagnostic tool. The incidence of cardiotoxicity attributed to treatment was 38.7%. The predictive power of cardiac troponin I alone was 58.3%, whereas it increases to 91% when combined with electrocardiography and to 95% when combined with echocardiographic study. The age, anthracyclines dose and the use of other chemotherapeutics increase the risk of anthracycline-induced cardiotoxicity. Cardiac troponin I is a simple non-invasive indicator for the presence of anthracycline-induced cardiotoxicity especially when used in combination with other parameters.


Risk of Clostridium difficile diarrhea among hospital inpatients prescribed proton pump inhibitors: cohort and case-control studies.
Antibiotic disruption of the normal intestinal flora is a well-known risk factor for Clostridium difficile-associated diarrhea. Reduced gastric acidity has been suggested as a risk factor, and we hypothesized that proton pump inhibitors, because of their potency, may be an independent risk factor for this problem. For the cohort study we identified from a pharmacy database 1187 inpatients at a Montreal teaching hospital who received antibiotics over a 9-month period beginning in August 2002. We compared patients in this group who had also received a proton pump inhibitor or an H(2) blocker with patients who had not received acid suppressive therapy. Hospital laboratory reports of positive assay results for C. difficile toxin were used to ascertain cases in the cohort. To assess the possibility that proton pump inhibitors were prescribed to patients who were sicker and had other risk factors for C. difficile infection, we did a case-control study at a second Montreal teaching hospital. Cases were defined as patients who were positive for C. difficile toxin and who had a history of diarrhea (n = 94). Control subjects were selected from among patients who had received an antibiotic and were matched to cases by ward, age within 5 years and class of antibiotics (n = 94). In the cohort study, C. difficile diarrhea developed in 81 (6.8%) of the 1187 patients who received antibiotics while in hospital. In a multivariate analysis, C. difficile diarrhea was significantly associated with use of proton pump inhibitors (adjusted odds ratio [OR] 2.1, 95% confidence interval [CI] 1.2- 3.5), receipt of 3 or more antibiotics (OR 2.1, 95% CI 1.3- 3.4) and admission to a medical ward (OR 4.1, 95% CI 2.3- 7.3). In the case-control study C. difficile diarrhea was associated with female sex (adjusted OR 2.1, 95% CI 1.1-4.0), prior renal failure (adjusted OR 4.3, 95% CI 1.5-11.9), hospital admission in the 3 months before the index admission (adjusted OR 2.6, 95% CI 1.4-5.2) and use of proton pump inhibitors (adjusted OR 2.7, 95% CI 1.4-5.2). Patients in hospital who received proton pump inhibitors were at increased risk of C. difficile diarrhea.


Effects of television modeling on residential energy conservation.
A combination of social marketing, communications, social learning (particularly modeling), and behavior analysis may provide an effective framework for behavior change via films and television. We used this approach in developing special television programs about residential energy conservation. The programs were tailored and directed to preselected middle-class homeowners (N = 150), and delivered over a public access channel of a cable TV system. The results indicated that after one program exposure (about 20 minutes), viewers adopted simple strategies modeled in the programs which led to savings of approximately 10% on their home energy use for a substantial part of the cooling and heating season. Although the potential benefits to costs of large-scale media efforts seemed great, institutional barriers for such programs were identified. Less expensive, more local programs seem more viable.


The professional and personal characteristics of effective psychotherapists: a systematic review.
Objective: Psychotherapists differ notably in the outcomes their patients achieve, and the characteristics that may explain these differences have attracted increasing interest. We systematically review studies on therapist pre-treatment characteristics predicting patient outcomes. Method: Systematic searches on databases for psychotherapy research, clinical psychology, and medical science for the years 2000-2018 identified published research examining therapist characteristics and psychotherapy outcomes. Of 2041 studies, 31 met inclusion criteria. Results: Findings show a few direct effects of therapist intrapersonal variables (e.g., self-relatedness, attachment) and several interaction effects with other constructs (e.g., patient pathology) on outcome. There is little support for the relevance of self-rated social skills. However, more consistent evidence has recently emerged for performance-based measurements of professional interpersonal skills, especially when elicited in challenging situations. Patient outcomes were also predicted by therapists' self-rated professional characteristics, such as their experienced difficulties in practice, coping mechanisms, and attitudes towards therapeutic work, indicating that therapist self-perception also matters, although not always in the direction expected. Conclusions: More effective therapists seem characterized by professionally cultivated interpersonal capacities, which are likely rooted in their personal lives and attachment history. Research guidelines are proposed for moving this field forward (including larger samples, multilevel modeling, and in-depth qualitative work).


Azide and trans-cyclooctene dUTPs: incorporation into DNA probes and fluorescent click-labelling.
5-Azidomethyl dUTP and two 5-trans-cyclooctene dUTPs with different linkers between the TCO and the uracil base have been incorporated into DNA by primer extension, reverse-transcription and PCR amplification. For azidomethyl dUTP the PCR reaction was successful even when the modified dUTP was not supplemented with dTTP. In one case 335 azidomethyl dU residues were incorporated into the 523 base pair amplicon using this methodology. 5-Azidomethyl dUTP was found to be a better substrate for DNA polymerases than the trans-cyclooctene dUTPs. However, the inverse electron demand Diels-Alder reaction between cyclooctene DNA and a tetrazine Cy3-dye was more efficient than the strain-promoted reaction between azide DNA and a bicyclo [6.1.0] non-4-yne Cy3 dye.


Differences in cause-specific infant mortality among Chinese, Japanese, and white Americans.
Although black-white differences in infant mortality have received much attention, information is limited about mortality differentials among Asian Americans. This study investigated racial differences in infant mortality in a sample of 21,288 Chinese, 11,882 Japanese, and 65,818 white resident singleton livebirths obtained from the National Center for Health Statistics 1983 and 1984 linked birth/infant death files. The crude infant mortality rates were 8.03, 6.56, and 8.46 per 1,000 livebirths for Chinese, Japanese, and white births, respectively. Cause-specific mortality varied considerably among the three racial groups. While the Japanese had lower rates of infant deaths and deaths from perinatal conditions for firstborn infants, they had higher rates of sudden infant death syndrome, as did Chinese females. The results of a logistic regression analysis indicate that the racial differences in total and cause-specific mortality persist when adjustment is made for demographic factors, use of prenatal care, infant sex, and birth weight. The effect of these latter variables on infant mortality varied by causes of death. The relations between infant mortality and variables such as marital status, maternal education, and birth interval appear indirect, operating partially through birth weight. While birth weight was the single strongest determinant of infant mortality, its relative importance varied by cause of death. The study findings suggest that policy decisions surrounding racial differences in infant mortality should not only be considered in light of specific races, but also with regard to cause-specific mortality. Moreover, additional research is needed to understand the cultural, biological, and behavioral factors that give rise to the racial differences.


Effects of ethanol ingestion on blood pressure reactivity.
We examined the acute effects of ethanol consumption on circulatory responses to the cold pressor test, to handgrip exercise and to intravenous infusion of methoxamine in eight normal male subjects. Ethanol consumption reduced systolic blood pressure responses to the cold pressor test and to handgrip exercise and depressed both systolic and diastolic blood pressure responses to infusion of methoxamine. The depressant effects of ethanol on blood pressure responses to methoxamine were considerably greater than the effects on the cold pressor test and handgrip exercise. Catecholamine concentrations during the cold pressor test and handgrip exercise were not significantly different with or without prior ethanol consumption. Alcohol has been shown to have an acute hypotensive action with all the stimuli.


Diatom aggregation when exposed to crude oil and chemical dispersant: Potential impacts of ocean acidification.
Diatoms play a key role in the marine carbon cycle with their high primary productivity and release of exudates such as extracellular polymeric substances (EPS) and transparent exopolymeric particles (TEP). These exudates contribute to aggregates (marine snow) that rapidly transport organic material to the seafloor, potentially capturing contaminants like petroleum components. Ocean acidification (OA) impacts marine organisms, especially those that utilize inorganic carbon for photosynthesis and EPS production. Here we investigated the response of the diatom Thalassiosira pseudonana grown to present day and future ocean conditions in the presence of a water accommodated fraction (WAF and OAWAF) of oil and a diluted chemically enhanced WAF (DCEWAF and OADCEWAF). T. pseudonana responded to WAF/DCEWAF but not OA and no multiplicative effect of the two factors (i.e., OA and oil/dispersant) was observed. T. pseudonana released more colloidal EPS (< 0.7 μm to > 3 kDa) in the presence of WAF/DCEWAF/OAWAF/OADCEWAF than in the corresponding Controls. Colloidal EPS and particulate EPS in the oil/dispersant treatments have higher protein-to-carbohydrate ratios than those in the control treatments, and thus are likely stickier and have a greater potential to form aggregates of marine oil snow. More TEP was produced in response to WAF than in Controls; OA did not influence its production. Polyaromatic hydrocarbon (PAH) concentrations and distributions were significantly impacted by the presence of dispersants but not OA. PAHs especially Phenanthrenes, Anthracenes, Chrysenes, Fluorenes, Fluoranthenes, Pyrenes, Dibenzothiophenes and 1-Methylphenanthrene show major variations in the aggregate and surrounding seawater fraction of oil and oil plus dispersant treatments. Studies like this add to the current knowledge of the combined effects of aggregation, marine snow formation, and the potential impacts of oil spills under ocean acidification scenarios.


Women's preference for receiving abortion in primary care settings.
Abortion is increasingly being incorporated into primary care training programs. It remains unclear if women, given the option, would accept and access these services with their primary care provider. An anonymous survey was administered to 299 women at two abortion clinics in New York and Chicago. One hundred seventy-four (58.2%) respondents theoretically would choose their primary care clinic for their abortion procedure, with more Chicago participants than New York participants choosing primary care. After adjusting for other variables, only being very or somewhat comfortable with one's primary care provider regarding issues of pregnancy prevention and birth control predicted the choice of primary care for abortion services. The majority of women surveyed theoretically would choose to have a procedure at their primary care clinic. There were some regional differences between New York and Chicago. Although many variables used in this study did not explain the respondents' preferences, the primary care clinic as site for abortion care appears to be an important potential option for many women. Primary care training programs should teach full-spectrum reproductive healthcare to help meet the needs of their patients.


Effect of staining and denture cleaning on color stability of differently polymerized denture base acrylic resins.
Denture hygiene is an important factor to remove dental plaque and discoloration, to provide optimal oral health, and to prevent denture stomatitis. The purpose of this study was to evaluate the effect of coffee staining and denture cleaner on the color stability of two differently polymerized (heat and microwave) denture acrylic resin in vitro. Two commonly used denture base acrylic resins and effervesecnt tablet form denture cleaner were used in this study. Disc-shaped specimens from each acrylic resin were polymerized according to the manufacturer's instructions (n = 7). Color values were recorded before and after immersion in distilled water (I), denture cleaner (II), coffee (III), and coffee + denture cleaner (IV) with a tristimulus colorimeter using CIEDE2000 color difference formula. Two-way analysis of variance (ANOVA) and Fisher's least significant difference tests were used for multiple comparisons at a 95% confidence level (P < 0.05). Surface topography of the acrylic resin denture base specimens before and after immersion were taken under 500 × magnification. The two-way ANOVA revealed significant interactions between denture base resins and solutions (P < 0.001). Immersion in denture cleaner and coffee solution caused significant color changes in acrylic resin denture bases. Heat- and microwave-polymerized acrylic denture base resins can show different color stability. Immersion in coffee and denture cleaner solutions can cause noticeable color changes.


Impaired hypocretin/orexin system alters responses to salient stimuli in obese male mice.
The brain has evolved in an environment where food sources are scarce, and foraging for food is one of the major challenges for survival of the individual and species. Basic and clinical studies show that obesity or overnutrition leads to overwhelming changes in the brain in animals and humans. However, the exact mechanisms underlying the consequences of excessive energy intake are not well understood. Neurons expressing the neuropeptide hypocretin/orexin (Hcrt) in the lateral/perifonical hypothalamus (LH) are critical for homeostatic regulation, reward seeking, stress response, and cognitive functions. In this study, we examined adaptations in Hcrt cells regulating behavioral responses to salient stimuli in diet-induced obese mice. Our results demonstrated changes in primary cilia, synaptic transmission and plasticity, cellular responses to neurotransmitters necessary for reward seeking, and stress responses in Hcrt neurons from obese mice. Activities of neuronal networks in the LH and hippocampus were impaired as a result of decreased hypocretinergic function. The weakened Hcrt system decreased reward seeking while altering responses to acute stress (stress-coping strategy), which were reversed by selectively activating Hcrt cells with chemogenetics. Taken together, our data suggest that a deficiency in Hcrt signaling may be a common cause of behavioral changes (such as lowered arousal, weakened reward seeking, and altered stress response) in obese animals.


Acetylcholine beyond bronchoconstriction: roles in inflammation and remodeling.
Acetylcholine is the primary parasympathetic neurotransmitter in the airways, where it not only induces bronchoconstriction and mucus secretion, but also regulates airway inflammation and remodeling. In this review, we propose that these effects are all primarily mediated via the muscarinic M3 receptor. Acetylcholine promotes inflammation and remodeling via direct effects on airway cells, and via mechanical stress applied to the airways sequential to bronchoconstriction. The effects on inflammation and remodeling are regulated by both neuronal and non-neuronal acetylcholine. Taken together, we believe that the combined effects of anticholinergic therapy on M3-mediated bronchoconstriction, mucus secretion, inflammation, and remodeling may account for the positive outcome of treatment with these drugs for patients with chronic pulmonary obstructive disease (COPD) or asthma.


Congenital laryngeal webs: surgical course and outcomes.
We compare the success of different surgical options in the treatment of laryngeal webs. We performed a retrospective study spanning the years 1980 to 2005. Eighteen patients were identified. The average age at diagnosis was 6 months (range, 1 day to 2.5 years). The presenting symptoms included weak cry, stridor, airway obstruction, and difficulty breathing. Associated cardiac defects consistent with the diagnosis of 22q-syndrome were present in 7 patients. Webs were classified as grade I (5 patients), grade II (2 patients), grade III (10 patients), or grade IV (1 patient) according to the Cohen classification. In 5 patients, only endoscopic lysis was required. The remaining 13 patients underwent open procedures; 9 patients in this group required tracheotomy. An average of 1.3 open airway procedures was necessary to achieve a decannulation rate of 89%. After operation, 34% of patients had residual webbing and 20% had a weak or aphonic voice. Management of laryngeal webs is dependent on the severity of airway obstruction. Grade I and II webs can be treated endoscopically; more severe laryngeal webs usually require tracheotomy and open airway reconstruction.


Location of a polypeptide sequence within the alpha-subunit of the acetylcholine receptor containing the cholinergic binding site.
Proteolytic fragments of the alpha-subunit of the acetylcholine receptor of Torpedo electric organ were generated by digestion with Staphylococcus aureus V8 protease, and their ability to bind alpha-bungarotoxin was assessed following resolution on polyacrylamide gels and transfer to nitrocellulose. The position of the smallest fragment (Mr = 17,000) with toxin-binding activity was located within the primary sequence of the alpha-subunit by isolation and chemical characterization. The amino acid sequence at its amino terminus is Val-Asn-Gln-Ile-Val-Glu, which is identical to a unique sequence on the alpha-subunit beginning at Val 46. Based on considerations of the apparent molecular weight of this polypeptide fragment, the enzyme specificity of V8 protease, analysis of the partial amino acid composition, and the position of various identifying landmarks of the primary sequence of the alpha-subunit, the carboxy terminus is restricted to an acidic amino acid residue between Asp 152 and Asp 180. The apparent affinities of the 17-kDa polypeptide for alpha-bungarotoxin (IC50 = 100 nM) and d-tubocurarine (IC50 = 0.4 mM) were not significantly different from the values obtained for the undigested alpha-subunit, suggesting that the 17-kDa polypeptide contains all of the essential elements present on the alpha-subunit that are necessary for the binding of both of these cholinergic ligands. A second toxin-binding fragment (Mr = 19,000) was also identified. Although the amino terminus of this polypeptide fragment has not been determined directly, its position relative to Asn 141, the site of N-glycosylation, was established, and it appears that its amino terminus cannot be located any closer to the amino terminus of the alpha-subunit than Asp 152. If this tentative assignment is correct, then the region of the alpha-subunit involved in binding cholinergic ligands is more closely defined as occurring within the segment of the primary sequence of the alpha-subunit that the 17- and 19-kDa polypeptide fragments have in common, i.e., an overlapping sequence maximally bounded by Asp 152 and Asp 180.


In vivo angiogenic activity of enamel matrix derivative.
Porcine enamel matrix derivative (EMD) has a clinical use in facilitating periodontal healing by enhancing the regeneration of alveolar bone, cementum, and periodontal ligament. The mechanism of clinical use has not been elucidated, but in vitro studies suggest that EMD may enhance healing, in part, by stimulating angiogenesis. This study analyzes the effect of EMD on the production of blood vessels in the chorioallantoic membrane (CAM) of the developing chicken egg. Various amounts of EMD ranging from 15 to 125 ng/5 microl were pipetted onto a 3-mm diameter x 2-mm thick bioabsorbable hemostatic gelatin and placed onto the surface of the CAM. Recombinant human amelogenin and a purified 5-kDa protein fraction derived from EMD were also tested at various amounts ranging from 15 to 62 ng/5 microl. A mixture of fibroblast growth factor and vascular endothelial growth factor served as the positive control. The negative control was 0.9% saline. A histologic examination of the interface of the gelatin and CAM was performed, evaluating for new blood-vessel formation on an ordinal scale of 0 to 3. Non-parametric statistical analyses were applied to compare test groups with the negative controls. CAM treated with EMD displayed moderate vascularity as indicated by a maximal score of 3.0 +/- 0.05 (mean +/- SEM). This compared favorably to the degree of vascularity stimulated by the mixture of fibroblast growth factor and vascular endothelial growth factor, which had a score of 3.0 +/- 0.05. Interestingly, the stimulation of angiogenesis by EMD was significant only at the lowest amounts tested. At the higher amounts, vascularity was reduced and not significantly different from the negative control. Vascularity was also increased by recombinant human amelogenin as indicated by a maximal score of 2.9 +/- 0.14. By contrast, there was only mild vascularity in sections treated with the negative control as indicated by a score of 1.7 +/- 0.4. The vascularity of the 5-kDa EMD-protein fraction was not different from the negative-control group (2.5 +/- 0.5 versus 1.7 +/- 0.4, respectively). EMD stimulates angiogenesis in the CAM model. As a heterogeneous mixture of extracellular matrix components, EMD may have multiple biologic functions, but it is likely that at least part of the explanation for its observed positive clinical effects may be the stimulation of angiogenesis. The fact that vascularity was also increased by recombinant human amelogenin raises the possibility that this 28.9-kDa protein may be the source of the angiogenic activity because it is the predominant protein of the EMD mixture. These results, taken together with results from previously reported in vitro studies, suggest that EMD may increase angiogenesis directly and/or indirectly at the wound-healing site.


The Australasian Resuscitation in Sepsis Evaluation (ARISE) trial statistical analysis plan.
The Australasian Resuscitation in Sepsis Evaluation (ARISE) study is an international, multicentre, randomised, controlled trial designed to evaluate the effectiveness of early goal-directed therapy compared with standard care for patients presenting to the emergency department with severe sepsis. In keeping with current practice, and considering aspects of trial design and reporting specific to non-pharmacological interventions, our plan outlines the principles and methods for analysing and reporting the trial results. The document is prepared before completion of recruitment into the ARISE study, without knowledge of the results of the interim analysis conducted by the data safety and monitoring committee and before completion of the two related international studies. Our statistical analysis plan was designed by the ARISE chief investigators, and reviewed and approved by the ARISE steering committee. We reviewed the data collected by the research team as specified in the study protocol and detailed in the study case report form. We describe information related to baseline characteristics, characteristics of delivery of the trial interventions, details of resuscitation, other related therapies and other relevant data with appropriate comparisons between groups. We define the primary, secondary and tertiary outcomes for the study, with description of the planned statistical analyses. We have developed a statistical analysis plan with a trial profile, mock-up tables and figures. We describe a plan for presenting baseline characteristics, microbiological and antibiotic therapy, details of the interventions, processes of care and concomitant therapies and adverse events. We describe the primary, secondary and tertiary outcomes with identification of subgroups to be analysed. We have developed a statistical analysis plan for the ARISE study, available in the public domain, before the completion of recruitment into the study. This will minimise analytical bias and conforms to current best practice in conducting clinical trials.


State-change decisions and dorsomedial prefrontal cortex: the importance of time.
Different kinds of decision making can be categorized by their differential effect on the agent's current and future states as well as the computational challenges they pose. Here, we draw a distinction between within-state and state-change decision-making, and propose that a dedicated decision mechanism exists in dorsomedial prefrontal cortex (dmPFC) that is specialized for state-change decisions. We set out a formal framework in which state change decisions may be made on the basis of the integrated momentary reward rate, over the intended time to be spent in a state. A key feature of this framework is that reward rate is expressed as a function of continuous time. We argue that dmPFC is suited for this type of decision making partly due to its ability to track the passage of time. This proposed function of dmPFC is placed in contrast to other evaluative systems such as the orbitofrontal cortex, which is important for careful deliberation within a specific model-space or option-space and within a decision strategy.


Brain anatomy differences in childhood stuttering.
Stuttering is a developmental speech disorder that occurs in 5% of children with spontaneous remission in approximately 70% of cases. Previous imaging studies in adults with persistent stuttering found left white matter deficiencies and reversed right-left asymmetries compared to fluent controls. We hypothesized that similar differences might be present indicating brain development differences in children at risk of stuttering. Optimized voxel-based morphometry compared gray matter volume (GMV) and diffusion tensor imaging measured fractional anisotropy (FA) in white matter tracts in 3 groups: children with persistent stuttering, children recovered from stuttering, and fluent peers. Both the persistent stuttering and recovered groups had reduced GMV from normal in speech-relevant regions: the left inferior frontal gyrus and bilateral temporal regions. Reduced FA was found in the left white matter tracts underlying the motor regions for face and larynx in the persistent stuttering group. Contrary to previous findings in adults who stutter, no increases were found in the right hemisphere speech regions in stuttering or recovered children and no differences in right-left asymmetries. Instead, a risk for childhood stuttering was associated with deficiencies in left gray matter volume while reduced white matter integrity in the left hemisphere speech system was associated with persistent stuttering. Anatomical increases in right hemisphere structures previously found in adults who stutter may have resulted from a lifetime of stuttering. These findings point to the importance of considering the role of neuroplasticity during development when studying persistent forms of developmental disorders in adults.


Upregulation of acid ceramidase contributes to tumor progression in tuberous sclerosis complex.
Tuberous sclerosis complex (TSC) is characterized by multisystem, low-grade neoplasia involving the lung, kidneys, brain, and heart. Lymphangioleiomyomatosis (LAM) is a progressive pulmonary disease affecting almost exclusively women. TSC and LAM are both caused by mutations in TSC1 and TSC2 that result in mTORC1 hyperactivation. Here, we report that single-cell RNA sequencing of LAM lungs identified activation of genes in the sphingolipid biosynthesis pathway. Accordingly, the expression of acid ceramidase (ASAH1) and dihydroceramide desaturase (DEGS1), key enzymes controlling sphingolipid and ceramide metabolism, was significantly increased in TSC2-null cells. TSC2 negatively regulated the biosynthesis of tumorigenic sphingolipids, and suppression of ASAH1 by shRNA or the inhibitor ARN14976 (17a) resulted in markedly decreased TSC2-null cell viability. In vivo, 17a significantly decreased the growth of TSC2-null cell-derived mouse xenografts and short-term lung colonization by TSC2-null cells. Combined rapamycin and 17a treatment synergistically inhibited renal cystadenoma growth in Tsc2+/- mice, consistent with increased ASAH1 expression and activity being rapamycin insensitive. Collectively, the present study identifies rapamycin-insensitive ASAH1 upregulation in TSC2-null cells and tumors and provides evidence that targeting aberrant sphingolipid biosynthesis pathways has potential therapeutic value in mechanistic target of rapamycin complex 1-hyperactive neoplasms, including TSC and LAM.


Influence of carrier lifetime on performance of silicon p-i-n variable optical attenuators fabricated on submicrometer rib waveguides.
We investigated influence of carrier lifetime on performance of silicon (Si) p-i-n variable optical attenuators (VOAs) on submicrometer Si rib waveguides. VOAs were fabricated with and without intentional implantation of lattice defects into their intrinsic region. Carrier lifetime was measured by pulse responses for normal incidence of picosecond laser pulse of 775 nm to the VOA, as approximately 1 ns and approximately 7 ns for the VOAs with and without defects, respectively. Carrier lifetime is determined by the sum of surface recombination and Auger recombination for VOAs without defects, while Schockley-Read-Hall recombination is dominant for the VOA with defects. As a result, attenuation efficiency (dB/mA) is 0.2-0.7 and 0.04-0.1, while 3-dB bandwidth is 40-100 MHz and over 200 MHz for the VOAs with and without defects, respectively. There is a trade-off relation between attenuation and response speed of the VOAs with respect to carrier lifetime i.e., attenuation efficiency is linearly proportional to the carrier lifetime, whereas response speed is inversely proportional to it.


An official European Respiratory Society/American Thoracic Society technical standard: field walking tests in chronic respiratory disease.
Field walking tests are commonly employed to evaluate exercise capacity, assess prognosis and evaluate treatment response in chronic respiratory diseases. In recent years, there has been a wealth of new literature pertinent to the conduct of the 6-min walk test (6MWT), and a growing evidence base describing the incremental and endurance shuttle walk tests (ISWT and ESWT, respectively). The aim of this document is to describe the standard operating procedures for the 6MWT, ISWT and ESWT, which can be consistently employed by clinicians and researchers. The Technical Standard was developed by a multidisciplinary and international group of clinicians and researchers with expertise in the application of field walking tests. The procedures are underpinned by a concurrent systematic review of literature relevant to measurement properties and test conduct in adults with chronic respiratory disease. Current data confirm that the 6MWT, ISWT and ESWT are valid, reliable and responsive to change with some interventions. However, results are sensitive to small changes in methodology. It is important that two tests are conducted for the 6MWT and ISWT. This Technical Standard for field walking tests reflects current evidence regarding procedures that should be used to achieve robust results.


When the ophthalmologists turn blind.
The cost of technology is high in ophthalmology but given the increasingly competitive environment and the social demand, there is a pressure to progressively lower the costs to the consumer. To keep costs down there is a tendency to do as many surgeries as possible in an assembly line fashion both in hospitals as well as in the charitable camps. This article provides ophthalmologists an insight into the legal pitfalls in practice of ophthalmology in India and the dangers of the constant lowering of costs of surgery as well as of free service. This lowering of costs would have been ideal in a Utopian world, but times have now changed and there is cost to be paid even for providing free service. In India the prevalent tradition of providing free service, has also resulted in a lowering of guard by the eye surgeons. These mass eye surgery assembly popularly called "free eye camps" has seen millions of people benefited. But recently there is an increase in number of cases where exorbitant penalty has been imposed by the courts, on these philanthropic surgeons for any deficiency in service, and this has destroyed the careers of many ophthalmologists. Time has now come to introspect and to factor the cost of litigation and compensations into the cost of surgeries so that we not only benefit the patients but also safeguard the ophthalmologists and help them fulfill their responsibilities towards their own dependents.


A cost-effectiveness analysis of in-vitro fertilization by maternal age and number of treatment attempts.
The increase in use and costs of assisted reproductive therapies including in-vitro fertilization (IVF) has led to debate over public funding. A decision analytic model was designed to estimate the incremental cost-effectiveness of IVF by additional treatment programmes and maternal age. Data from the Australian and New Zealand Assisted Reproductive Database were used to estimate incremental effects (live birth and other pregnancy outcomes) and costs for cohorts of women attempting up to three treatment programmes. A treatment programme included one fresh cycle and a variable number of frozen cycles dependent on maternal age. The incremental cost per live birth ranged from AU dollars 27 373 and AU dollars 31 986 for women aged 30-33 on their first and third programmes to AU dollars 130 951 and AU dollars 187 515 for 42-45-year-old women on their first and second attempts. Overall, these trends were not affected by inclusions of costs associated with ovarian hyperstimulation syndrome or multiple births. This study suggests that cost per live birth from IVF increases with maternal age and treatment programme number and indicates that maternal age has the much greater effect. This evidence may help decisionmakers target the use of IVF services conditional on societal willingness to pay for live births and equity considerations.


Nucleotide receptor-mediated decrease of tight-junctional permeability in cultured human cervical epithelium.
Extracellular ATP changes the transepithelial electrical conductance (GT) across cultures of human cervical cells acutely, in a biphasic manner that is characterized by a rapid increase (phase I) followed by a sustained decrease in GT (phase II). We tested the hypothesis that the phase II response is mediated by decreases in the permeability of tight junctions. We studied the effect of ATP on the relative mobilities of Cl- vs. Na+ (uCl/uNa) as calculated from changes in the dilution potential (Vdil). Vdil was induced by lowering NaCl from 130 to 10 mM in either the luminal or subluminal solutions bathing filters containing cells. uCl/uNa was 1.27 across cervical cultures and 1.34 across blank filters, compared with a level of 1.52 in free solution. Increases in GT induced by transepithelial hydrostatic or hypertonic gradients (which increase permeability of lateral intercellular space) had no effect on uCl/uNa. Increases in GT induced by lowering extracellular Ca2+ to < 0.1 mM increased uCl/uNa to levels obtained in blank filters, indicating abrogation of tight-junctional resistance. Phase I response and ionomycin (which produces a sustained phase I-like increase in GT) had no effect on uCl/uNa. The phase II response, however, decreased uCl/uNa from 1.27 to 1.24, and the effect could be abrogated by lowering extracellular Ca2+. These results indicate that phase II decreases in GT across cultured human cervical epithelium are mediated by acute decreases in tight-junctional permeability.


College of American Pathologists' laboratory standards for next-generation sequencing clinical tests.
The higher throughput and lower per-base cost of next-generation sequencing (NGS) as compared to Sanger sequencing has led to its rapid adoption in clinical testing. The number of laboratories offering NGS-based tests has also grown considerably in the past few years, despite the fact that specific Clinical Laboratory Improvement Amendments of 1988/College of American Pathologists (CAP) laboratory standards had not yet been developed to regulate this technology. To develop a checklist for clinical testing using NGS technology that sets standards for the analytic wet bench process and for bioinformatics or "dry bench" analyses. As NGS-based clinical tests are new to diagnostic testing and are of much greater complexity than traditional Sanger sequencing-based tests, there is an urgent need to develop new regulatory standards for laboratories offering these tests. To develop the necessary regulatory framework for NGS and to facilitate appropriate adoption of this technology for clinical testing, CAP formed a committee in 2011, the NGS Work Group, to deliberate upon the contents to be included in the checklist. Results . -A total of 18 laboratory accreditation checklist requirements for the analytic wet bench process and bioinformatics analysis processes have been included within CAP's molecular pathology checklist (MOL). This report describes the important issues considered by the CAP committee during the development of the new checklist requirements, which address documentation, validation, quality assurance, confirmatory testing, exception logs, monitoring of upgrades, variant interpretation and reporting, incidental findings, data storage, version traceability, and data transfer confidentiality.


Challenges from Tuberculosis Diagnosis to Care in Community-Based Active Case Finding among the Urban Poor in Cambodia: A Mixed-Methods Study.
While community-based active case finding (ACF) for tuberculosis (TB) holds promise for increasing early case detection among hard-to-reach populations, limited data exist on the acceptability of active screening. We aimed to identify barriers and explore facilitators on the pathway from diagnosis to care among TB patients and health providers. Mixed-methods study. We administered a survey questionnaire to, and performed in-depth interviews with, TB patients identified through ACF from poor urban settlements in Phnom Penh, Cambodia. Additionally, we conducted focus group discussions and in-depth interviews with community and public health providers involved in ACF, respectively. Acceptance of home TB screening was strong among key stakeholders due to perceived reductions in access barriers and in direct and indirect patient costs. Privacy and stigma were not an issue. To build trust and facilitate communication, the participation of community representatives alongside health workers was preferred. Most health providers saw ACF as complementary to existing TB services; however, additional workload as a result of ACF was perceived as straining operating capacity at public sector sites. Proximity to a health facility and disease severity were the strongest determinants of prompt care-seeking. The main reasons reported for delays in treatment-seeking were non-acceptance of diagnosis, high indirect costs related to lost income/productivity and transportation expenses, and anticipated side-effects from TB drugs. TB patients and health providers considered home-based ACF complementary to facility-based TB screening. Strong engagement with community representatives was believed critical in gaining access to high risk communities. The main barriers to prompt treatment uptake in ACF were refusal of diagnosis, high indirect costs, and anticipated treatment side-effects. A patient-centred approach and community involvement were essential in mitigating barriers to care in marginalised communities.


Disease-causing missense mutations within the N-terminal transmembrane domain of GlcNAc-1-phosphotransferase impair endoplasmic reticulum translocation or Golgi retention.
Transport of newly synthesized lysosomal enzymes to the lysosome requires tagging of these enzymes with the mannose 6-phosphate moiety by UDP-GlcNAc:lysosomal enzyme N-acetylglucosamine-1-phosphotransferase (GlcNAc-1-phosphotransferase), encoded by two genes, GNPTAB and GNPTG. GNPTAB encodes the α and β subunits, which are initially synthesized as a single precursor that is cleaved by Site-1 protease in the Golgi. Mutations in this gene cause the lysosomal storage disorders mucolipidosis II (MLII) and mucolipidosis III αβ (MLIII αβ). Two recent studies have reported the first patient mutations within the N-terminal transmembrane domain (TMD) of the α subunit of GlcNAc-1-phosphotransferase that cause either MLII or MLIII αβ. Here, we demonstrate that two of the MLII missense mutations, c.80T>A (p.Val27Asp) and c.83T>A (p.Val28Asp), prevent the cotranslational insertion of the nascent GlcNAc-1-phosphotransferase polypeptide chain into the endoplasmic reticulum. The remaining four mutations, one of which is associated with MLII, c.100G>C (p.Ala34Pro), and the other three with MLIII αβ, c.70T>G (p.Phe24Val), c.77G>A (p.Gly26Asp), and c.107A>C (p.Glu36Pro), impair retention of the catalytically active enzyme in the Golgi with concomitant mistargeting to endosomes/lysosomes. Our results uncover the basis for the disease phenotypes of these patient mutations and establish the N-terminal TMD of GlcNAc-1-phosphotransferase as an important determinant of Golgi localization.


State of the art of Ready-to-Use Therapeutic Food: a tool for nutraceuticals addition to foodstuff.
Therapeutic foodstuff are a challenge for the use of food and functional food ingredients in the therapy of different pathologies. Ready-to-Use Therapeutic Food (RUTF) are a mixture of nutrients designed and primarily addressed to the therapy of the severe acute malnutrition. The main ingredients of the formulation are powdered milk, peanuts butter, vegetal oil, sugar, and a mix of vitamins, salts, and minerals. The potential of this food are the low percentage of free water and the high energy and nutritional density. The high cost of the powdered milk, and the food safety problems connected to the onset of toxigenic moulds on the peanuts butter, slowed down considerably the widespread and homogenous diffusion of this product. This paper presents the state of the art of RUTF, reviews the different proposed recipes, suggests some possible new formulations as an alternative of novel recipes for this promising food.


Pd2+ fluorescent sensors based on amino and imino derivatives of rhodamine and improvement of water solubility by the formation of inclusion complexes with β-cyclodextrin.
Mono- and bis-rhodamine derivatives appended with amino (RhB1) and imino (BRI) groups, respectively, have been designed as colorimetric and fluorescent sensors for the selective detection of Pd2+. In addition, an attempt has been made to improve the water solubility of the probe by synthesizing inclusion complexes of RhB1 with β-cyclodextrin as the host molecule. The resulting probe, RhB1-CD, exhibited exactly the same physicochemical phenomena as the guest molecule RhB1, indicating that complexation with β-CD improved the water solubility of RhB1 without affecting its sensing ability. With increasing concentrations of Pd2+, the absorption (556 nm) and emission (591 nm) intensities of BRI as well as the absorption intensities of RhB1/RhB1-CD (563 nm) increase, whereas the emission intensities of RhB1/RhB1-CD decrease due to fluorescence quenching. Therefore, BRI can act as an "OFF-ON" fluorescent probe switch in CH3CN/H2O (3 : 2 v/v) with a detection limit of 1 μM in solution, whereas RhB1 and RhB1-CD act as "ON-OFF" fluorescent probes in EtOH/H2O (1 : 1 v/v) and water, respectively, with a detection limit of 10 μM. All these probes exhibited high selectivity for Pd2+ and reversibility by treatment with Na2S. In the presence of commonly coexisting metal ions, such as Hg2+, Fe3+, Cu2+ and Al3+, BRI undergoes either catalytic Schiff base hydrolysis or it is unaffected by its affinity to the metal ions. However, Pd2+ can be effectively distinguished by a 12-15 nm bathochromic shift in its emission spectrum when compared with that of other metal ions. A Job's plot revealed that BRI forms 1 : 2 complexes by a spirolactam ring opening mechanism, whereas RhB1/RhB1-CD form 2 : 1 complexes by a ring opening mechanism followed by Pd2+-induced dimerization; the proposed binding mechanisms are presented.


First report of the in vitro nematicidal effects of camel milk.
Antipathogenic properties of camel milk have been investigated to substitute for drugs hence overcome drug resistance. The main objective of this present study was to investigate the anthelmintic activity of camel milk in relationship to its chemical composition. In vitro anthelmintic effects of camel milk against Haemonchus contortus from sheep were ascertained by egg hatching and worm motility inhibitions in comparison to milks from cow, ewe and goat as well as a reference drug albendazole. Chemical composition revealed that camel milk has higher contents of protective protein (lactoferrin) and vitamin C than other species' milk. It showed ovicidal activity at all tested concentrations and completely inhibited egg hatching at a concentration close to 100mg/mL (inhibitory concentration (IC50)=42.39mg/mL). Camel milk revealed in vitro activity against adult parasites in terms of the paralysis and/or death of the worms at different hours post treatment. After 8h of exposure, it induced 100% mortality at the highest tested concentration. There was 82.3% immobility of worms in albendazole 8h post-exposition. No such effects were seen with the other species' milks. Bioactive compounds such as lactoferrin and vitamin C may be involved in such an effect. To our knowledge, these results depict for the first time that camel milk possesses in vitro anthelmintic properties and further in vitro and in vivo trials against different parasite species and stages are required to make use of this milk for the control of gastrointestinal nematode parasites.


Hepatitis B surface antigen vector delivers protective cytotoxic T-lymphocyte responses to disease-relevant foreign epitopes.
Although hepatitis B surface antigen (HBsAg) per se is highly immunogenic, its use as a vector for the delivery of foreign cytotoxic T-lymphocyte (CTL) epitopes has met with little success because of constraints on HBsAg stability and secretion imposed by the insertion of foreign sequence into critical hydrophobic/amphipathic regions. Using a strategy entailing deletion of DNA encoding HBsAg-specific CTL epitopes and replacement with DNA encoding foreign CTL epitopes, we have derived chimeric HBsAg DNA immunogens which elicited effector and memory CTL responses in vitro, and pathogen- and tumor-protective responses in vivo, when the chimeric HBsAg DNAs were used to immunize mice. We further show that HBsAg DNA recombinant for both respiratory syncytial virus and human papillomavirus CTL epitopes elicited simultaneous responses to both pathogens. These data demonstrate the efficacy of HBsAg DNA as a vector for the delivery of disease-relevant protective CTL responses. They also suggest the applicability of the approach of deriving chimeric HBsAg DNA immunogens simultaneously encoding protective CTL epitopes for multiple diseases. The DNAs we tested formed chimeric HBsAg virus-like particles (VLPs). Thus, our results have implications for the development of vaccination strategies using either chimeric HBsAg DNA or VLP vaccines. HBsAg is the globally administered vaccine for hepatitis B virus infection, inviting its usage as a vector for the delivery of immunogens from other diseases.


Biodiversity, community structural shifts, and biogeography of prokaryotes within Antarctic continental shelf sediment.
16S ribosomal DNA (rDNA) clone library analysis was conducted to assess prokaryotic diversity and community structural changes within a surficial sediment core obtained from an Antarctic continental shelf area (depth, 761 m) within the Mertz Glacier Polynya (MGP) region. Libraries were created from three separate horizons of the core (0- to 0.4-cm, 1.5- to 2.5-cm, and 20- to 21-cm depth positions). The results indicated that at the oxic sediment surface (depth, 0 to 0.4 cm) the microbial community appeared to be dominated by a small subset of potentially r-strategist (fast-growing, opportunistic) species, resulting in a lower-than-expected species richness of 442 operational taxonomic units (OTUs). At a depth of 1.5 to 2.5 cm, the species richness (1,128 OTUs) was much higher, with the community dominated by numerous gamma and delta proteobacterial phylotypes. At a depth of 20 to 21 cm, a clear decline in species richness (541 OTUs) occurred, accompanied by a larger number of more phylogenetically divergent phylotypes and a decline in the predominance of Proteobacteria. Based on rRNA and clonal abundance as well as sequence comparisons, syntrophic cycling of oxidized and reduced sulfur compounds appeared to be the dominant process in surficial MGP sediment, as phylotype groups putatively linked to these processes made up a large proportion of clones throughout the core. Between 18 and 65% of 16S rDNA phylotypes detected in a wide range of coastal and open ocean sediments possessed high levels of sequence similarity (>95%) with the MGP sediment phylotypes, indicating that many sediment prokaryote phylotype groups defined in this study are ubiquitous in marine sediment.


European domestic horses originated in two holocene refugia.
The role of European wild horses in horse domestication is poorly understood. While the fossil record for wild horses in Europe prior to horse domestication is scarce, there have been suggestions that wild populations from various European regions might have contributed to the gene pool of domestic horses. To distinguish between regions where domestic populations are mainly descended from local wild stock and those where horses were largely imported, we investigated patterns of genetic diversity in 24 European horse breeds typed at 12 microsatellite loci. The distribution of high levels of genetic diversity in Europe coincides with the distribution of predominantly open landscapes prior to domestication, as suggested by simulation-based vegetation reconstructions, with breeds from Iberia and the Caspian Sea region having significantly higher genetic diversity than breeds from central Europe and the UK, which were largely forested at the time the first domestic horses appear there. Our results suggest that not only the Eastern steppes, but also the Iberian Peninsula provided refugia for wild horses in the Holocene, and that the genetic contribution of these wild populations to local domestic stock may have been considerable. In contrast, the consistently low levels of diversity in central Europe and the UK suggest that domestic horses in these regions largely derive from horses that were imported from the Eastern refugium, the Iberian refugium, or both.


Floral structure and development in Rafflesiaceae with emphasis on their exceptional gynoecia.
The holoparasitic plant family Rafflesiaceae include the world's largest flowers. Despite their iconic status, relatively little is known about the morphology and development of their flowers. A recent study clarified the organization of the outer (sterile) floral organs, surprisingly revealing that their distinctive floral chambers arose via different developmental pathways in the two major genera of the family. Here, we expand that research to investigate the structure and development of the reproductive organs of Rafflesiaceae. Serial sectioning, scanning electron microscopy, and x-ray tomography of floral buds were employed to reconstruct the structure and development of all three Rafflesiaceae genera. Unlike most angiosperms, which form their shoot apex from the primary morphological surface, the shoot apex of Rafflesiaceae instead forms secondarily via internal cell separation (schizogeny) along the distal boundary of the host-parasite interface. Similarly, the radially directed ovarial clefts of the gynoecium forms via schizogeny within solid tissue, and no carpels are initiated from the floral apex. The development of the shoot apex and gynoecium of Rafflesiaceae are highly unusual. Although secondary formation of the morphological surface from the shoot apex has been documented in other plant groups, secondary derivation of the inner gynoecium surface is otherwise unknown. Both features are likely synapomorphies of Rafflesiaceae. The secondary derivation of the shoot apex may protect the developing floral shoot as it emerges from within dense host tissue. The secondary formation of the ovarial clefts may generate the extensive placental area necessary to produce hundreds of thousands of ovules.


Interleukine-2 therapy does not increase the risk of Hodgkin or non-Hodgkin lymphoma in HIV-infected patients: results from FHDH ANRS CO4.
Concerns have been raised about a possible excess risk of lymphomas in HIV-infected patients exposed to interleukin 2 (IL-2) therapy. Here we compared the risks of non-Hodgkin lymphoma (NHL) and Hodgkin lymphoma (HL) in IL-2-treated and IL-2-untreated HIV-infected patients. Patients monitored through the French Hospital Database on HIV between May 1, 1995, and December 31, 2005, were enrolled in this study. Lymphomas that occurred between the day after study entry and the end of follow-up were eligible for analysis. Poisson regression models were used in 2 separate analyses to quantify the possible relationship between IL-2 therapy and the incidence of NHL and HL. The IL-2-treated group consisted of 861 patients and the IL-2-untreated group of 77,605 patients. Follow-up lasted a total of 3643 and 382,720 person-years, respectively. After adjustment for sex and time-updated age, period, the CD4 cell counts, the plasma HIV RNA levels, and AIDS status, the relative rates of NHL and HL associated with IL-2 therapy were 0.64 (95% confidence interval, 0.25 to 1.65) and 0.33 (95% confidence interval, 0.04 to 2.86), respectively. In this large observational study, IL-2 therapy did not increase the risk of lymphoma, either NHL or HL, in HIV-infected patients.


Stress and reproductive hormones reflect inter-specific social and nutritional conditions mediated by resource availability in a bear-salmon system.
Food availability can influence the nutritional and social dynamics within and among species. Our investigation focused on grizzly and black bears in coastal British Columbia, Canada, where recent and dramatic declines in their primary prey (salmon) raise concerns about potentially negative effects on bear physiology. We examined how salmon availability relates to stress and reproductive hormones in coastal grizzly (n = 69) and black bears (n = 68) using cortisol and testosterone. In hair samples from genotyped individuals, we quantified salmon consumption using stable isotope analysis and hormone levels by enzyme immunoassay. To estimate the salmon biomass available to each bear, we developed a spatially explicit approach based on typical bear home-range sizes. Next, we compared the relative importance of salmon consumption and salmon availability on hormone levels in male bears using an information theoretical approach. Cortisol in grizzly bears was higher in individuals that consumed less salmon, possibly reflecting nutritional stress. In black bears, cortisol was better predicted by salmon availability than salmon consumption; specifically, individuals in areas and years with low salmon availability showed higher cortisol levels. This indicates that cortisol in black bears is more strongly influenced by the socially competitive environment mediated by salmon availability than by nutritional requirements. In both species, testosterone generally decreased with increasing salmon availability, possibly reflecting a less competitive environment when salmon were abundant. Differences between species could relate to different nutritional requirements, social densities and competitive behaviour and/or habitat use. We present a conceptual model to inform further investigations in this and other systems. Our approach, which combines data on multiple hormones with dietary and spatial information corresponding to the year of hair growth, provides a promising tool for evaluating the responses of a broad spectrum of wildlife to changes in food availability or other environmental conditions.


Evidence that both normal and immune elimination of Schistosoma mansoni take place at the lung stage of migration prior to parasite death.
The number and distribution of autoradiographic foci observed in this and previous studies following percutaneous infection with 75Se-labeled Schistosoma mansoni cercariae indicate that the lungs are the principal site of worm elimination in both normal mice and mice immunized with irradiated cercariae. It was observed in the present study, however, that the intensities of the autoradiographic foci produced in the lungs during both the normal (early) and immune (late) phases of elimination were identical to those of foci produced in the livers of the same mice by larvae shown to be alive. In contrast, foci produced in the lungs by heat-killed, intravenously injected, lung schistosomula became smaller and fainter with time, disappearing completely between seven and 10 days after injection in normal mice and between four and six days in immunized mice. These results indicate that although the targets of both normal and immune elimination do not proceed beyond the lung stage of migration, they do not die in the lungs. A possible explanation for this paradoxical situation, for which there is some experimental evidence, is that unsuccessful migrators leave the blood stream, enter alveoli, pass up the trachea, and are eventually digested in the gastrointestinal tract or eliminated from the body intact.


Heparin regulates the collagen phenotype of vascular smooth muscle cells: induced synthesis of an Mr 60,000 collagen.
The effect of heparin on the biosynthetic phenotype of rat vascular smooth muscle cells (SMC) was investigated in vitro. Addition of heparin to the culture medium of early passage rat SMC resulted in a marked (3-15-fold) increase of a cell layer-associated Mr 60,000 protein that was sensitive to digestion by purified bacterial collagenase and contained significant amounts of hydroxyproline. Pulse-chase analysis of heparin-treated SMC revealed that the Mr 60,000 collagen was a primary and abundant product of these cells and was not processed extracellularly to a smaller form. The inductive effect of heparin could be mimicked by iota carrageenan or dextran sulfates but not by hyaluronic acid, dermatan sulfate, or chondroitin sulfates. The induction was concentration dependent with a maximal effect observed at a heparin concentration of 10 micrograms/ml. Synthesis of the Mr 60,000 collagen increased 18-24 h after addition of heparin to the cultures. Following induction and subsequent removal of heparin, synthesis of the protein remained maximal for at least 12 h and required 72 h to return to a basal level. These data demonstrate that the biosynthetic phenotype of vascular SMC in vitro can be controlled, at least in part, by heparin and related polyanions and suggest a role for similar molecules endogenous to the vessel wall in the regulation of SMC function.


Notch ligand Dll4 impairs cell recruitment to aortic clusters and limits blood stem cell generation.
Hematopoietic stem cells (HSCs) develop from the hemogenic endothelium in cluster structures that protrude into the embryonic aortic lumen. Although much is known about the molecular characteristics of the developing hematopoietic cells, we lack a complete understanding of their origin and the three-dimensional organization of the niche. Here, we use advanced live imaging techniques of organotypic slice cultures, clonal analysis, and mathematical modeling to show the two-step process of intra-aortic hematopoietic cluster (IACH) formation. First, a hemogenic progenitor buds up from the endothelium and undergoes division forming the monoclonal core of the IAHC. Next, surrounding hemogenic cells are recruited into the IAHC, increasing their size and heterogeneity. We identified the Notch ligand Dll4 as a negative regulator of the recruitment phase of IAHC. Blocking of Dll4 promotes the entrance of new hemogenic Gfi1+ cells into the IAHC and increases the number of cells that acquire HSC activity. Mathematical modeling based on our data provides estimation of the cluster lifetime and the average recruitment time of hemogenic cells to the cluster under physiologic and Dll4-inhibited conditions.


Virtual Exploration of the Ring Systems Chemical Universe.
Here, we explore the chemical space of all virtually possible organic molecules focusing on ring systems, which represent the cyclic cores of organic molecules obtained by removing all acyclic bonds and converting all remaining atoms to carbon. This approach circumvents the combinatorial explosion encountered when enumerating the molecules themselves. We report the chemical universe database GDB4c containing 916 130 ring systems up to four saturated or aromatic rings and maximum ring size of 14 atoms and GDB4c3D containing the corresponding 6 555 929 stereoisomers. Almost all (98.6%) of these ring systems are unknown and represent chiral 3D-shaped macrocycles containing small rings and quaternary centers reminiscent of polycyclic natural products. We envision that GDB4c can serve to select new ring systems from which to design analogs of such natural products. The database is available for download at www.gdb.unibe.ch together with interactive visualization and search tools as a resource for molecular design.


Principles and application of educational counseling used in progressive audiologic tinnitus management.
Exposure to loud sounds is a common cause and exacerbater of tinnitus - a troubling auditory symptom that affects millions of people worldwide. Clinical research at the National Center for Rehabilitative Auditory Research has resulted in a clinical model of tinnitus management referred to as Progressive Audiologic Tinnitus Management (PATM). The model involves five hierarchical levels of management: Triage, Audiologic Evaluation, Group Education, Tinnitus Evaluation, and Individualized Management. Counseling by audiologists and, as needed, mental health providers, is a key component of PATM. This style of counseling focuses less on didactic informational counseling; instead, counseling is used for facilitating patients' learning to adjust to the disturbing auditory symptom by successfully employing tools from two powerful skillsets for self-management of chronic tinnitus - the therapeutic uses of sound and techniques from cognitive-behavioral psychology. This article provides an overview of the methods of counseling used with PATM and provides details concerning the overarching principles of collaborative adult learning that are believed to be most important in facilitating self-management by patients who complain of tinnitus.


Intermittent preventive treatment in infants for the prevention of malaria in rural Western kenya: a randomized, double-blind placebo-controlled trial.
Intermittent preventive treatment in infants (IPTi) with sulphadoxine-pyrimethamine (SP) for the prevention of malaria has shown promising results in six trials. However, resistance to SP is rising and alternative drug combinations need to be evaluated to better understand the role of treatment versus prophylactic effects. Between March 2004 and March 2008, in an area of western Kenya with year round malaria transmission with high seasonal intensity and high usage of insecticide-treated nets, we conducted a randomized, double-blind placebo-controlled trial with SP plus 3 days of artesunate (SP-AS3), 3 days of amodiaquine-artesunate (AQ3-AS3), or 3 days of short-acting chlorproguanil-dapsone (CD3) administered at routine expanded programme of immunization visits (10 weeks, 14 weeks and 9 months). 1,365 subjects were included in the analysis. The incidence of first or only episode of clinical malaria during the first year of life (primary endpoint) was 0.98 episodes/person-year in the placebo group, 0.74 in the SP-AS3 group, 0.76 in the AQ3-AS3 group, and 0.82 in the CD3 group. The protective efficacy (PE) and 95% confidence intervals against the primary endpoint were: 25.7% (6.3, 41.1); 25.9% (6.8, 41.0); and 16.3% (-5.2, 33.5) in the SP-AS3, AQ3-AS3, and CD3 groups, respectively. The PEs for moderate-to-severe anaemia were: 27.5% (-6.9, 50.8); 23.1% (-11.9, 47.2); and 11.4% (-28.6, 39.0). The duration of the protective effect remained significant for up to 5 to 8 weeks for SP-AS3 and AQ3-AS3. There was no evidence for a sustained beneficial or rebound effect in the second year of life. All regimens were well tolerated. These results support the view that IPTi with long-acting regimens provide protection against clinical malaria for up to 8 weeks even in the presence of high ITN coverage, and that the prophylactic rather than the treatment effect of IPTi appears central to its protective efficacy.


Soft governance, restratification and the 2004 general medical services contract: the case of UK primary care organisations and general practice teams.
In the UK National Health Service, primary care organisation (PCO) managers have traditionally relied on the soft leadership of general practitioners based on professional self-regulation rather than direct managerial control. The 2004 general medical services contract (nGMS) represented a significant break from this arrangement by introducing new performance management mechanisms for PCO managers to measure and improve general practice work. This article examines the impact of nGMS on the governance of UK general practice by PCO managers through a qualitative analysis of data from an empirical study in four UK PCOs and eight general practices, drawing on Hood's four-part governance framework. Two hybrids emerged: (i) PCO managers emphasised a hybrid of oversight, competition (comptrol) and peer-based mutuality by granting increased support, guidance and autonomy to compliant practices; and (ii) practices emphasised a broad acceptance of increased PCO oversight of clinical work that incorporated a restratified elite of general practice clinical peers at both PCO and practice levels. Given the increased international focus on the quality, safety and efficiency in primary care, a key issue for PCOs and practices will be to achieve an effective, contextually appropriate balance between the counterposing governance mechanisms of peer-led mutuality and externally led comptrol.


Polarization-independent active metamaterial for high-frequency terahertz modulation.
We present a polarization-independent metamaterial design for the construction of electrically tunable terahertz (THz) devices. The implemented structure consists of an array of gold crosses fabricated on top of an n-doped gallium arsenide (GaAs) layer. Utilizing THz time-domain spectroscopy, we show that the electric resonance and thus the transmission properties of the cross structure can be tuned by an externally applied bias voltage. We further demonstrate the fast amplitude modulation of a propagating THz wave for modulation frequencies up to 100 kHz.