Hemogenic Reprogramming of Human Fibroblasts by Enforced Expression of Transcription Factors.
The cellular and molecular mechanisms underlying specification of human hematopoietic stem cells (HSCs) remain elusive. Strategies to recapitulate human HSC emergence in vitro are required to overcome limitations in studying this complex developmental process. Here, we describe a protocol to generate hematopoietic stem and progenitor-like cells from human dermal fibroblasts employing a direct cell reprogramming approach. These cells transit through a hemogenic intermediate cell-type, resembling the endothelial-to-hematopoietic transition (EHT) characteristic of HSC specification. Fibroblasts were reprogrammed to hemogenic cells via transduction with GATA2, GFI1B and FOS transcription factors. This combination of three factors induced morphological changes, expression of hemogenic and hematopoietic markers and dynamic EHT transcriptional programs. Reprogrammed cells generate hematopoietic progeny and repopulate immunodeficient mice for three months. This protocol can be adapted towards the mechanistic dissection of the human EHT process as exemplified here by defining GATA2 targets during the early phases of reprogramming. Thus, human hemogenic reprogramming provides a simple and tractable approach to identify novel markers and regulators of human HSC emergence. In the future, faithful induction of hemogenic fate in fibroblasts may lead to the generation of patient-specific HSCs for transplantation.


Mutations of Arabidopsis in potential transduction and response components of the phototropic signaling pathway.
Four genetic loci were recently identified by mutations that affect phototropism in Arabidopsis thaliana (L.) Heyhn. seedlings. It was hypothesized that one of these loci, NPH1, encodes the apoprotein for a phototropic photoreceptor. All of the alleles at the other three mutant loci (nph2, nph3, and nph4) contained wild-type levels of the putative NPH1 protein and exhibited normal blue-light-dependent phosphorylation of the NPH1 protein. This indicated that the NPH2, NPH3, and NPH4 proteins likely function downstream of NPH1 photoactivation. We show here that, although the nph2, nph3, and nph4 mutants are all altered with respect to their phototropic responses, only the nph4 mutants are also altered in their gravitropic responsiveness. Thus, NPH2 and NPH3 appear to act as signal carriers in a phototropism-specific pathway, whereas NPH4 is required for both phototropism and gravitropism and thus may function directly in the differential growth response. Despite their altered phototropic responses in blue and green light as etiolated seedlings, the nph2 and nph4 mutants exhibited less dramatic mutant phenotypes as de-etiolated seedlings and when etiolated seedlings were irradiated with unilateral ultraviolet-A (UV-A) light. Examination of the phototropic responses of a mutant deficient in biologically active phytochromes, hy1-100, indicated that phytochrome transformation by UV-A light mediates an increase in phototropic responsiveness, accounting for the greater phototropic curvature of the nph2 and nph4 mutants to UV-A light than to blue light.


Canine parvovirus serology: a collaborative assay.
Fifteen laboratories were supplied with coded samples of canine sera for testing for the presence of antibodies against canine parvoviruses. One of these sera had been designated as a potential British standard canine parvovirus antiserum. Most of these laboratories were either providing a canine parvovirus serology service, or represented pharmaceutical companies which manufacture canine parvovirus vaccines for the United Kingdom market. No attempt was made to influence the test methods used. Thirteen of the laboratories used a haemagglutination inhibition test, three an enzyme-linked immunosorbent assay (ELISA), and two performed serum neutralisation tests. Three laboratories used two different techniques. Adequate analysis was possible only with the results of the haemagglutination inhibition tests. The variability of the results between laboratories could be partly controlled by the use of the standard serum. Much of the residual variability was associated with particular laboratories. The results from the vaccine manufacturers tended to be less variable than those from the diagnostic laboratories.


GDNF: a glial cell line-derived neurotrophic factor for midbrain dopaminergic neurons.
A potent neurotrophic factor that enhances survival of midbrain dopaminergic neurons was purified and cloned. Glial cell line-derived neurotrophic factor (GDNF) is a glycosylated, disulfide-bonded homodimer that is a distantly related member of the transforming growth factor-beta superfamily. In embryonic midbrain cultures, recombinant human GDNF promoted the survival and morphological differentiation of dopaminergic neurons and increased their high-affinity dopamine uptake. These effects were relatively specific; GDNF did not increase total neuron or astrocyte numbers nor did it increase transmitter uptake by gamma-aminobutyric-containing and serotonergic neurons. GDNF may have utility in the treatment of Parkinson's disease, which is marked by progressive degeneration of midbrain dopaminergic neurons.


Plasma carotenoid levels are not directly related to heavy metal exposure or reproductive success in three insectivorous passerines.
The authors measured plasma carotenoid levels in three insectivorous bird species, the pied flycatcher (Ficedula hypoleuca), the blue tit (Cyanistes caeruleus), and the great tit (Parus major) in metal-polluted and unpolluted sites around a copper smelter. Their aim was to determine whether there was interspecific, age-related, or yearly variation in carotenoid levels and their responses to the ambient pollution level. The three bird species showed qualitatively and quantitatively similar carotenoid profiles, with lutein being always the predominant plasma carotenoid. Ficedula hypoleuca nestlings showed higher plasma carotenoid levels in the unpolluted than in the polluted area, whereas no significant differences were found in F. hypoleuca females or in nestlings or females of the Parids. Yearly comparison of plasma concentrations (only for P. major nestlings) showed, however, that levels varied nonsynchronously between polluted and unpolluted sites among the breeding seasons, with the yearly variation being higher in the polluted area. The survival of nestlings did not depend on their plasma carotenoid concentrations. The two Parid species showed lower nestling survival at sites with high metal exposure levels. The authors conclude that plasma carotenoid levels showed a marked spatial, temporal, and age-related variation, but were not directly related to heavy metal exposure or the reproductive success of birds.


Non-targeted metabolomics in sport and exercise science.
Metabolomics incorporates the study of metabolites that are produced and released through physiological processes at both the systemic and cellular levels. Biological compounds at the metabolite level are of paramount interest in the sport and exercise sciences, although research in this field has rarely been referred to with the global 'omics terminology. Commonly studied metabolites in exercise science are notably within cellular pathways for adenosine triphosphate production such as glycolysis (e.g., pyruvate and lactate), β-oxidation of free fatty acids (e.g., palmitate) and ketone bodies (e.g., β-hydroxybutyrate). Non-targeted metabolomic technologies are able to simultaneously analyse the large numbers of metabolites present in human biological samples such as plasma, urine and saliva. These analytical technologies predominately employ nuclear magnetic resonance spectroscopy and chromatography coupled to mass spectrometry. Performing experiments based on non-targeted methods allows for systemic metabolite changes to be analysed and compared to a particular physiological state (e.g., pre-/post-exercise) and provides an opportunity to prospect for metabolite signatures that offer beneficial information for translation into an exercise science context, for both elite performance and public health monitoring. This narrative review provides an introduction to non-targeted metabolomic technologies and discusses current and potential applications in sport and exercise science.


Alcohol assessment & feedback by e-mail for university student hazardous and harmful drinkers: study protocol for the AMADEUS-2 randomised controlled trial.
Alcohol is responsible for a large and growing proportion of the global burden of disease, as well as being the cause of social problems. Brief interventions are one component of comprehensive policy measures necessary to reduce these harms. Brief interventions increasingly take advantage of the Internet to reach large numbers of high risk groups such as students. The research literature on the efficacy and effectiveness of online interventions is developing rapidly. Although many studies show benefits in the form of reduced consumption, other intervention studies show no effects, for reasons that are unclear. Sweden became the first country in the world to implement a national system in which all university students are offered a brief online intervention via an e-mail. This randomized controlled trial (RCT) aims to evaluate the effectiveness of this national system comprising a brief online intervention among university students who are hazardous and harmful drinkers. This study employs a conventional RCT design in which screening to determine eligibility precedes random allocation to immediate or delayed access to online intervention. The online intervention evaluated comprises three main components; assessment, normative feedback and advice on reducing drinking. Screening is confined to a single question in order to minimise assessment reactivity and to prevent contamination. Outcomes will be evaluated after 2 months, with total weekly alcohol consumption being the primary outcome measure. Invitations to participate are provided by e-mail to approximately 55,000 students in 9 Swedish universities. This RCT evaluates routine service provision in Swedish universities via a delay in offer of intervention to the control group. It evaluates effects in the key population for whom this intervention has been designed. Study findings will inform the further development of the national service provision. ISRCTN02335307.


A qualitative study of senior hospital managers' views on current and innovative strategies to improve hand hygiene.
Despite universal recognition of the importance of hand hygiene in reducing the incidence of healthcare associated infections, health care workers' compliance with best practice has been sub-optimal. Senior hospital managers have responsibilities for implementing patient safety initiatives and are therefore ideally placed to provide suggestions for improving strategies to increase hand hygiene compliance. This is an under-researched area, accordingly the aim of this study was to identify senior hospital managers' views on current and innovative strategies to improve hand hygiene compliance. Qualitative design comprising face-to-face interviews with thirteen purposively sampled senior managers at a major teaching and referral hospital in Sydney, Australia. Data were analysed thematically. Seven themes emerged: culture change starts with leaders, refresh and renew the message, connect the five moments to the whole patient journey, actionable audit results, empower patients, reconceptualising non-compliance and start using the hammer. To strengthen hand hygiene programmes, strategies based on the five moments of hand hygiene should be tailored to specific roles and settings and take into account the whole patient journey including patient interactions with clinical and non-clinical staff. Senior clinical and non-clinical leaders should visibly champion and mandate best practice initiatives and articulate that hand hygiene non-compliance is culturally and professionally unacceptable to the organization. Strategies that included a disciplinary component and which conceptualise hand hygiene non-compliance as a patient safety error may be worth evaluating in terms of staff acceptability and effectiveness.


Comparison of total ionic strength adjustment buffers III and IV in the measurement of fluoride concentration of teas.
Tea is the second most consumed drink in the UK and a primary source of hydration; it is an important source of dietary fluoride (F) for consumers and also abundant in aluminium (Al). Varying ranges of F concentrations in teas have been reported worldwide which may be, in part, due to differences in analytical techniques used to measure this ion. The effect of using total ionic adjustment buffers (TISAB) III or IV when measuring F concentration of black teas available in the UK was investigated and compared. Based on this evaluation, the effects of three different infusion times, 1 min, 10 min and 1 h, caffeine content and tea form on the F contents of the tea samples were investigated. The F concentrations of 47 tea samples were measured directly using a fluoride ion-selective electrode (F-ISE), TISAB III and IV and infusion times of 1 min, 10 min and 1 h. Mean (SD) F concentration of tea samples for all infusion times was statistically significantly higher ( p < 0.001) measured by TISAB IV (4.37 (2.16) mg/l) compared with TISAB III (3.54 (1.65) mg/l). A statistically significant positive correlation ( p < 0.001) was found between Al concentration (mg/l) and differences in F concentration (mg/l) measured using the two TISABs; the difference in F concentration measured by the two TISABs increased with the magnitude of Al concentration. Due to higher concentrations of F and Al in teas and their complexing potential, use of TISAB IV facilitates more accurate measurement of F concentration when using an F-ISE and a direct method.


Polarized Raman spectroscopy to elucidate the texture of synthesized MoS2.
Texture has a significant impact on several key properties of transition-metal dichalcogenides (TMDs) films. Films with in-plane oriented grains have been successfully implemented in nano- and opto-electronic devices, whereas, films with out-of-plane oriented material have shown excellent performance in catalytic applications. It will be demonstrated that the texture of nanocrystalline TMD films can be determined with polarized Raman spectroscopy. A model describing the impact of texture on the Raman response of 2D-TMDs will be presented. For the specific case of MoS2, the model was used to quantify the impact of texture on the relative strength of the A1g and E12g modes in both the unpolarized and polarized Raman configuration. Subsequently, the capability to characterize texture by polarized Raman was demonstrated on various MoS2 films grown by atomic-layer deposition (ALD) and validated by complementary transmission electron microscopy (TEM) and synchrotron based 2D grazing-incidence X-ray diffraction (GIXD) measurements. This also revealed how the texture evolved during ALD growth of MoS2 and subsequent annealing of the films. The insights presented in this work allow a deeper understanding of Raman spectra of nanocrystalline TMDs and enable a rapid and non-destructive method to probe texture.


Flaxseed sprouts induce apoptosis and inhibit growth in MCF-7 and MDA-MB-231 human breast cancer cells.
Flaxseeds have been shown to play a role in the prevention of cancer and heart disease, and it is believed that their more favorable fatty acid composition is responsible. Sprouting is a natural method to modify nutritional components and to decrease cyanide poisoning of raw flaxseeds. Here, we investigated the in vitro effects of flaxseed sprouts on cell growth and apoptosis of human breast cancer cells. In a series of in vitro experiments, estrogen-receptor-positive (MCF-7) and estrogen-receptor-negative (MDA-MB-231) cells were cultured and treated with flaxseed sprouts, and then cell proliferation, apoptosis, and gene expression were measured. Flaxseed sprouts significantly reduced the growth of both of MCF-7 and MDA-MB-231 cells and also increased apoptosis. However, flaxseed sprouts did not affect the growth of MCF-10A mammary epithelial cells. In gene transcription analysis using quantitative real-time polymerase chain reaction, flaxseed sprout treatment significantly upregulated p53 mRNA in both cell cancer lines. These results suggest that flaxseed sprouts induce apoptosis and inhibit cancer cell growth, thereby demonstrating their anti-proliferative effects in breast cancer cells. This study may provide important information for devising dietary strategies to reduce breast cancer risk.


Effects of Cone Connexin-36 Disruption on Light Adaptation and Circadian Regulation of the Photopic ERG.
The present study tested the hypothesis that connexin-36 (Cx36) and gap junctions between photoreceptor cells contribute to the circadian rhythm of the photopic electroretinogram (ERG) b-wave amplitude. Cone-specific disruption of Cx36 was obtained in mice with a floxed Gjd2 gene and human red/green pigment promoter (HRGP)-driven Cre recombinase. Standard ERG, spectral-domain optical coherence tomography (SD-OCT) and histochemical methods were used. HRGPcreGjd2fl/fl mice had a selective reduction in Cx36 protein in the outer plexiform layer; no reduction in Cx36 was observed in the inner plexiform layer. Cx36 disruption had no effect on the number of cones, the thickness of the photoreceptor layer, or the scotopic ERG responses. However, there was a reduction of the photopic ERG circadian rhythm, with b-wave amplitudes in the day and the night locked in the daytime, light-adapted state. In HRGPcreGjd2+/+and Gjd2fl/fl controls, the circadian rhythm of light-adapted ERG persisted, similar to that in wild type mice. Cx36 regulation contributes to the circadian rhythm of light-adapted ERG; in the absence of photoreceptor gap junctions, mice appear to be in a fully light-adapted state regardless of the time of day. The higher amplitudes and reduced circadian regulation of the b-wave of HRGPcreGjd2fl/fl mice may be due to increased synaptic strength at the cone to ON bipolar cell synapse due to electrotonic isolation of the terminals lacking gap junctions.


Dynamics of viremia in primary HIV-1 infection in Africans: insights from analyses of host and viral correlates.
In HIV-1 infection, plasma viral load (VL) has dual implications for pathogenesis and public health. Based on well-known patterns of HIV-1 evolution and immune escape, we hypothesized that VL is an evolving quantitative trait that depends heavily on duration of infection (DOI), demographic features, human leukocyte antigen (HLA) genotypes and viral characteristics. Prospective data from 421 African seroconverters with at least four eligible visits did show relatively steady VL beyond 3 months of untreated infection, but host and viral factors independently associated with cross-sectional and longitudinal VL often varied by analytical approaches and sliding time windows. Specifically, the effects of age, HLA-B(⁎)53 and infecting HIV-1 subtypes (A1, C and others) on VL were either sporadic or highly sensitive to time windows. These observations were strengthened by the addition of 111 seroconverters with 2-3 eligible VL results, suggesting that DOI should be a critical parameter in epidemiological and clinical studies.


Absence of a difference in the neurosecretory activity of supraoptic nucleus vasopressin neurons of neuroleptic-treated schizophrenic patients.
Dysfunction in water intake and metabolism has frequently been reported in schizophrenia. The general population of schizophrenics under neuroleptic treatment secretes lower amounts of vasopressin than controls at comparable values of plasma osmolality. The purpose of the present study was to investigate the synthetic activity of vasopressin neurons of the dorsolateral supraoptic nucleus in schizophrenia on postmortem material using a battery of histochemical activity markers. Our material consisted of formalin-fixed and paraffin-embedded hypothalami from 5 schizophrenic patients under neuroleptic treatment and from 5 matched controls, obtained from The Netherlands' Brain Bank. DSM-III or DSM-IV criteria were used for the clinical diagnosis. The histochemical markers used to study the neuronal activity of the magnocellular vasopressin-synthesizing neurons were: cell size, size of the Golgi apparatus, and expression of vasopressin and tyrosine hydroxylase mRNA by in situ hybridization. Morphometric evaluation and statistical analysis (Mann-Whitney U test) were performed. Our results showed no statistically significant differences in any of the neuronal activity markers between schizophrenic patients and controls. Therefore, the neurosecretory activity of vasopressin neurons of the dorsolateral part of the supraoptic nucleus does not appear to be changed in schizophrenic patients under medication. Since our sample did not include patients with reported polydipsia or hyponatremia, prospective investigation is needed to evaluate the above-mentioned neuronal activity markers in such a particular subgroup of schizophrenic patients.


A novel motif in the yeast mitochondrial dynamin Dnm1 is essential for adaptor binding and membrane recruitment.
To initiate mitochondrial fission, dynamin-related proteins (DRPs) must bind specific adaptors on the outer mitochondrial membrane. The structural features underlying this interaction are poorly understood. Using yeast as a model, we show that the Insert B domain of the Dnm1 guanosine triphosphatase (a DRP) contains a novel motif required for association with the mitochondrial adaptor Mdv1. Mutation of this conserved motif specifically disrupted Dnm1-Mdv1 interactions, blocking Dnm1 recruitment and mitochondrial fission. Suppressor mutations in Mdv1 that restored Dnm1-Mdv1 interactions and fission identified potential protein-binding interfaces on the Mdv1 β-propeller domain. These results define the first known function for Insert B in DRP-adaptor interactions. Based on the variability of Insert B sequences and adaptor proteins, we propose that Insert B domains and mitochondrial adaptors have coevolved to meet the unique requirements for mitochondrial fission of different organisms.


An immunotherapy survivor population: health-related quality of life and toxicity in patients with metastatic melanoma treated with immune checkpoint inhibitors.
The immune checkpoint inhibitors (ICIs) have resulted in subgroups of patients with metastatic melanoma achieving high-quality durable responses. Metastatic melanoma survivors are a new population in the era of cancer survivorship. The aim of this study was to evaluate metastatic melanoma survivors in terms of health-related quality of life (HRQoL), immune-related adverse events (irAEs) and exposure to immunosuppressive agents in a large single centre in the UK. We defined the survivor population as patients with a diagnosis of metastatic melanoma who achieved a durable response to an ICI and had been followed-up for a minimum of 12 months from initiation of ICI without disease progression. HRQoL was assessed using SF-36. Electronic health records were accessed to collect data on demographics, treatments, irAEs and survival. HRQoL data was compared with two norm-based datasets. Eighty-four metastatic melanoma survivors were eligible and 87% (N = 73) completed the SF-36. ICI-related toxicity of any grade occurred in 92% of patients and 43% had experienced a grade 3 or 4 toxicity. Almost half (49%) of the patients required steroids for the treatment of ICI-related toxicity, whilst 14% required treatment with an immunosuppressive agent beyond steroids. Melanoma survivors had statistically significant lower HRQoL scores with regard to physical, social and physical role functioning and general health compared with the normative population. There was a trend towards inferior scores in patients with previous exposure to ipilimumab compared with those never exposed to ipilimumab. Our results show that metastatic melanoma survivors have potentially experienced significant ICI-related toxicity and experience significant impairments in specific HRQoL domains. Future service planning is required to meet this population's unique survivorship needs.


Differences in Electronic Personal Health Information Tool Use Between Rural and Urban Cancer Patients in the United States: Secondary Data Analysis.
Studies have previously shown that rural cancer patients are diagnosed at later stages of disease. This delay is felt throughout treatment and follow-up, reflected in the fact that rural patients often have poorer clinical outcomes compared with their urban counterparts. Few studies have explored whether there is a difference in cancer patients' current use of health information technology tools by residential location. Data from 7 cycles of the Health Information National Trends Survey (HINTS, 2003-2017) were merged and analyzed to examine whether differences exist in managing electronic personal health information (ePHI) and emailing health care providers among rural and urban cancer patients. Geographic location was categorized using Rural-Urban Continuum Codes (RUCCs). Bivariate analyses and multivariable logistic regression were used to determine whether associations existed between rural/urban residency and use of health information technology among cancer patients. Of the 3031 cancer patients/survivors who responded across the 7 cycles of HINTS, 797 (26.9%) resided in rural areas. No difference was found between rural and urban cancer patients in having managed ePHI in the past 12 months (OR 0.78, 95% CI 0.43-1.40). Rural cancer patients were significantly less likely to email health care providers than their urban counterparts (OR 0.52, 95% CI 0.32-0.84). The digital divide between rural and urban cancer residents does not extend to general ePHI management; however, electronic communication with providers is significantly lower among rural cancer patients than urban cancer patients. Further research is needed to determine whether such disparities extend to other health information technology tools that might benefit rural cancer patients as well as other chronic conditions.


Cysteine-Rich Atrial Secretory Protein from the Snail Achatina achatina: Purification and Structural Characterization.
Despite extensive studies of cardiac bioactive peptides and their functions in molluscs, soluble proteins expressed in the heart and secreted into the circulation have not yet been reported. In this study, we describe an 18.1-kDa, cysteine-rich atrial secretory protein (CRASP) isolated from the terrestrial snail Achatina achatina that has no detectable sequence similarity to any known protein or nucleotide sequence. CRASP is an acidic, 158-residue, N-glycosylated protein composed of eight alpha-helical segments stabilized with five disulphide bonds. A combination of fold recognition algorithms and ab initio folding predicted that CRASP adopts an all-alpha, right-handed superhelical fold. CRASP is most strongly expressed in the atrium in secretory atrial granular cells, and substantial amounts of CRASP are released from the heart upon nerve stimulation. CRASP is detected in the haemolymph of intact animals at nanomolar concentrations. CRASP is the first secretory protein expressed in molluscan atrium to be reported. We propose that CRASP is an example of a taxonomically restricted gene that might be responsible for adaptations specific for terrestrial pulmonates.


Relationship between perceived stigma and depression severity.
The purpose of this study was to explore the relationship between perceived stigma and being in treatment for depression and current depression severity. Face-to-face interviews were conducted with a convenience sample of depressed subjects from a Veterans Administration outpatient mental health clinic (N = 54) and never-depressed subjects from a Veterans Administration primary care clinic (N = 50). Depression severity was measured using the 9-item Primary Care Evaluation of Mental Disorders depression measure. Stigma was measured using the 5-item Stigma Scale for Receiving Psychological Help modified for depression treatment. Statistical analyses included Spearman correlation and multivariate regression. In the correlation analysis, being in treatment for depression compared with never experiencing depression was associated with significantly higher levels of perceived stigma (p <.001). In separate multivariate models controlling for significant univariate correlates, greater depression severity (p <.001) and meeting criteria for current major depression (p <.001) were significant predictors of perceived stigma. Greater depression severity appears to be a strong predictor of perceived stigma.


Effect of mechanical cleaning of endotracheal tubes with sterile urethral catheters to reduce biofilm formation in ventilator patients.
To investigate the effectiveness of mechanical cleaning with sterile urethral catheters to prevent formation of biofilms on endotracheal tubes. Forty-five children were randomized in equal numbers to endotracheal tube cleaning group for three times a day (group A), twice daily (group B), or to a control group with no endotracheal tube cleaning (group C). Bacterial studies and confocal laser scanning microscopy were performed to assess bacterial colonization and biofilm thickness on the internal surface of the endotracheal tube. In group B, the quantities of viable bacteria adhering to the endotracheal tube after 1 day of ventilation were similar to the control group but were significantly less by 3 days (p < 0.05). The quantities of viable bacteria adhering to the endotracheal tube in group A were significantly lower than group C from day 1 to day 7 (p < 0.05). The numbers of culture-positive endotracheal tube and lower respiratory tract secretions were both reduced in groups A and B compared with group C. Confocal laser scanning microscopy showed progressive development of mature biofilms in group C. Scattered bacteria were seen in group A with no biofilm formation. In group B, a small amount of extracellular polymeric substance was seen, with more bacterial cells than in group A. The biofilms in group B were significantly thinner than those in group C (p < 0.05). The occurrence of ventilator-associated pneumonia was significantly reduced by endotracheal tube cleaning. Mechanical cleaning with sterile urethral catheters reduced bacterial colonization, prevented formation of endotracheal tube biofilm, and reduced the occurrence of ventilator-associated pneumonia.


Formation and closure of microchannels in skin following microporation.
To characterize the microchannels created in hairless rat skin by microneedles and investigate their closure following exposure to different occlusive conditions. Maltose microneedles were characterized by scanning electron microscopy. The microchannels created and their closure when exposed to different conditions was investigated using a variety of techniques. Microscopic imaging indicates a pyramidal geometry of maltose microneedles with an average length of 559 ± 14 μm and tip radius of 4 μm. Upon insertion into skin, they created microchannels with an average surface diameter of 60 μm and an average depth of 160 ± 20 μm as observed by histological sectioning and confocal microscopy. Skin recovers its barrier function within 3-4 hrs, and microchannels closed within 15 hrs of poration when exposed to environment. However, when occluded, the microchannels remained open for up to 72 hrs in vivo, as observed by calcein imaging, transepidermal water loss measurements and methylene blue staining. Maltose microneedles penetrated the stratum corneum barrier and created microchannels in skin which completely close within 15 hrs after poration. However, under occluded conditions, barrier recovery can be delayed for up to 72 hrs in vivo.


Cloak/anti-cloak interactions.
Coordinate-transformation cloaking is based on the design of a metamaterial shell made of an anisotropic, spatially inhomogeneous "transformation medium" that allows rerouting the impinging wave around a given region of space. In its original version, it is generally believed that, in the ideal limit, the radiation cannot penetrate the cloaking shell (from outside to inside, and viceversa). However, it was recently shown by Chen et al. that electromagnetic fields may actually penetrate the cloaked region, provided that this region contains double-negative transformation media which, via proper design, may be in principle used to (partially or totally) "undo" the cloaking transformation, thereby acting as an "anti-cloak." In this paper, we further elaborate this concept, by considering a more general scenario of cloak/anti-cloak interactions. Our full-wave analytical study provides new insightful results and explores the effects of departure from ideality, suggesting also some novel scenarios for potential applications.


Hybrid Nanomaterials of Conjugated Polymers and Albumin for Precise Photothermal Therapy.
Heretofore, conjugated polymers (CPs) attract considerable attention in photothermal therapy (PTT). Although various CPs with different structures have been reported, the suboptimal circulation persistence and biodistribution limit their efficacy in tumor treatment. Human serum albumin (HSA), an endogenous plasma protein, has been widely functioned as a carrier for therapeutic agents. Herein, we construct nanocomplex C16 pBDP@HSA nanoparticles (NPs) from hydrophobic 4,4-difluoro-4-bora-3 a,4 a-diaza- s-indacene (BODIPY)-containing CPs and HSA, which exhibit robust stability in physiological conditions and excellent photothermal activity upon irradiation. The high photothermal conversion efficiency of 37.5%, higher than that of other reported PTT agents such as gold nanorods, phosphorus quantum dots, and 2D materials, results in the potent photocytotoxicity toward cancer cells. Simultaneously, C16 pBDP@HSA NPs' capabilities of near-infrared fluorescence and photoacoustic imaging can provide guidance to the PTT. The outstanding inhibition of tumor growth results from great photothermal activity, the benefited accumulation in tumor, and optimal timing of treatment. To the best of our knowledge, this is the first study which combines the BODIPY-based CPs and HSA in one nanostructure and finds application in cancer treatment. Moreover, this article also offers a new strategy for other insoluble macromolecules to explore more biomedical applications.


Cognitive ontologies for neuropsychiatric phenomics research.
Now that genome-wide association studies (GWAS) are dominating the landscape of genetic research on neuropsychiatric syndromes, investigators are being faced with complexity on an unprecedented scale. It is now clear that phenomics, the systematic study of phenotypes on a genome-wide scale, comprises a rate-limiting step on the road to genomic discovery. To gain traction on the myriad paths leading from genomic variation to syndromal manifestations, informatics strategies must be deployed to navigate increasingly broad domains of knowledge and help researchers find the most important signals. The success of the Gene Ontology project suggests the potential benefits of developing schemata to represent higher levels of phenotypic expression. Challenges in cognitive ontology development include the lack of formal definitions of key concepts and relations among entities, the inconsistent use of terminology across investigators and time, and the fact that relations among cognitive concepts are not likely to be well represented by simple hierarchical "tree" structures. Because cognitive concept labels are labile, there is a need to represent empirical findings at the cognitive test indicator level. This level of description has greater consistency, and benefits from operational definitions of its concepts and relations to quantitative data. Considering cognitive test indicators as the foundation of cognitive ontologies carries several implications, including the likely utility of cognitive task taxonomies. The concept of cognitive "test speciation" is introduced to mark the evolution of paradigms sufficiently unique that their results cannot be "mated" productively with others in meta-analysis. Several projects have been initiated to develop cognitive ontologies at the Consortium for Neuropsychiatric Phenomics (www.phenomics.ucla.edu), in the hope that these ultimately will enable more effective collaboration, and facilitate connections of information about cognitive phenotypes to other levels of biological knowledge. Several free web applications are available already to support examination and visualisation of cognitive concepts in the literature (PubGraph, PubAtlas, PubBrain) and to aid collaborative development of cognitive ontologies (Phenowiki and the Cognitive Atlas). It is hoped that these tools will help formalise inference about cognitive concepts in behavioural and neuroimaging studies, and facilitate discovery of the genetic bases of both healthy cognition and cognitive disorders.


F-cell world drive 2011: are tactical medicine principles applicable to a civilian scenario?
In 2011, a Mercedes Benz (MB) conducted the F-Cell World Drive tour around the globe in 125 days. While crossing Asia from Shanghai (China) to Helsinki (Finland) by car, en route medical care was provided by embedded emergency physicians. The designated route crossed four different countries, multiple climate zones, and challenging road conditions. There was only limited information provided about hospitals and emergency medical services within different host nations in the planning phase, so we adopted tactical medical principles for mission planning and execution, as we were facing remote conditions and limitations to equipment, staffing, and patient transport.


Utilizing van der Waals Slippery Interfaces to Enhance the Electrochemical Stability of Silicon Film Anodes in Lithium-Ion Batteries.
High specific capacity anode materials such as silicon (Si) are increasingly being explored for next-generation, high performance lithium (Li)-ion batteries. In this context, Si films are advantageous compared to Si nanoparticle based anodes since in films the free volume between nanoparticles is eliminated, resulting in very high volumetric energy density. However, Si undergoes volume expansion (contraction) under lithiation (delithiation) of up to 300%. This large volume expansion leads to stress build-up at the interface between the Si film and the current collector, leading to delamination of Si from the surface of the current collector. To prevent this, adhesion promotors (such as chromium interlayers) are often used to strengthen the interface between the Si and the current collector. Here, we show that such approaches are in fact counter-productive and that far better electrochemical stability can be obtained by engineering a van der Waals "slippery" interface between the Si film and the current collector. This can be accomplished by simply coating the current collector surface with graphene sheets. For such an interface, the Si film slips with respect to the current collector under lithiation/delithiation, while retaining electrical contact with the current collector. Molecular dynamics simulations indicate (i) less stress build-up and (ii) less stress "cycling" on a van der Waals slippery substrate as opposed to a fixed interface. Electrochemical testing confirms more stable performance and much higher Coulombic efficiency for Si films deposited on graphene-coated nickel (i.e., slippery interface) as compared to conventional nickel current collectors.


Are specialist certification examinations a reliable measure of physician competence?
High stakes postgraduate specialist certification examinations have considerable implications for the future careers of examinees. Medical colleges and professional boards have a social and professional responsibility to ensure their fitness for purpose. To date there is a paucity of published data about the reliability of specialist certification examinations and objective methods for improvement. Such data are needed to improve current assessment practices and sustain the international credibility of specialist certification processes. To determine the component and composite reliability of the Fellowship examination of the College of Physicians of South Africa, and identify strategies for further improvement, generalizability and multivariate generalizability theory were used to estimate the reliability of examination subcomponents and the overall reliability of the composite examination. Decision studies were used to identify strategies for improving the composition of the examination. Reliability coefficients of the component subtests ranged from 0.58 to 0.64. The composite reliability of the examination was 0.72. This could be increased to 0.8 by weighting all test components equally or increasing the number of patient encounters in the clinical component of the examination. Correlations between examination components were high, suggesting that similar parameters of competence were being assessed. This composite certification examination, if equally weighted, achieved an overall reliability sufficient for high stakes examination purposes. Increasing the weighting of the clinical component decreased the reliability. This could be rectified by increasing the number of patient encounters in the examination. Practical ways of achieving this are suggested.


Effect of OSA on hypoxic and inflammatory markers during CPAP withdrawal: Further evidence from three randomized control trials.
Obstructive sleep apnoea (OSA) is associated with cardiovascular disease. Intermittent hypoxia, endothelial dysfunction and adipose tissue-mediated inflammation have all been linked to cardiovascular disease in OSA. We therefore explored the effect of OSA on relevant associated blood markers: adrenomedullin (ADM), endocan, endothelin-1 (ET-1), resistin and vascular endothelial growth factor (VEGF). Patients with OSA, established on and compliant with continuous positive airways pressure (CPAP) therapy for >1 year were included from three randomized controlled trials, conducted at two centres. Patients were randomized to either continued therapeutic CPAP or sham CPAP (CPAP withdrawal) for 2 weeks. Blood markers were measured at baseline and at 14 days and the treatment effect between sham CPAP and therapeutic CPAP was analysed. A total of 109 patients were studied (therapeutic CPAP n = 54, sham CPAP n = 55). Sham CPAP was associated with a return of OSA (between-group difference in oxygen desaturation index (ODI) 36.0/h, 95% CI 29.9-42.2, P < 0.001). Sham CPAP was associated with a reduction in ADM levels at 14 days (-26.0 pg/mL, 95% CI -47.8 to -4.3, P = 0.02), compared to therapeutic CPAP. Return of OSA was not associated with changes in endocan, ET-1, resistin or VEGF. Whilst CPAP withdrawal was associated with return of OSA, it was associated with an unexpected significant reduction in the vasodilator ADM and not with expected increases in hypoxia-induced markers, markers of endothelial function or resistin. We propose that the vascular effects occurring in OSA may be brought about by other mechanisms, perhaps partly through a reduction in ADM.


Genetic Diversity through the Looking Glass: Effect of Enrichment Bias.
The effect of enrichment bias on the diversity of 2,4-dichlorophenoxyacetate (2,4-D)-degrading (2,4-D(sup+)) bacteria recovered from soil was evaluated by comparing the diversity of isolates obtained by direct plating to the diversity of isolates obtained from 85 liquid batch cultures. By the two methods, a total of 159 isolates were purified from 1 g of soil and divided into populations based on repeated extragenic palindromic sequence PCR (rep-PCR) genomic fingerprints. Approximately 42% of the direct-plating isolates hybridized with the tfdA and tfdB genes from Alcaligenes eutrophus JMP134(pJP4), 27% hybridized with the tfdA and tfdB genes from Burkholderia sp. strain RASC, and 30% hybridized with none of the probes. In contrast, the enrichment isolates not only represented fewer populations than the isolates obtained by direct plating but also exhibited, almost exclusively, a single hybridization pattern with 2,4-D catabolic gene probes. Approximately 98% of the enrichment isolates possessed pJP4-type tfdA and tfdB genes, whereas isolates containing RASC-type tfdA and tfdB genes were obtained from only 2 of the 85 enrichment cultures. The skewed occurrence of the pJP4-type genes among the isolates obtained by enrichment suggests that the competitive fitness of 2,4-D(sup+) populations during growth with 2,4-D may be influenced either by specific tfd alleles or by genetic factors linked to these alleles. Moreover, the results indicate that evaluation of the diversity and distribution of catabolic pathways in nature can be highly distorted by the use of enrichment culture techniques.


Stereotactic injection of adenoviral vectors that target gene expression to specific pituitary cell types: implications for gene therapy.
Gene therapy is a potentially useful strategy for the treatment of pituitary adenomas or hormone deficiency disorders. We investigated the feasibility of targeting gene expression to specific pituitary cell types in vivo, using a combination of stereotactic injection and adenoviral vectors that carry pituitary-specific promoters. Recombinant adenoviruses containing the human growth hormone promoter (AdGHGal) or the human glycoprotein hormone alpha-subunit promoter (AdalphaGal) were used to drive expression of the beta-galactosidase gene. The expression of beta-galactosidase activity in the pituitary was analyzed after the administration of recombinant adenoviruses via the peripheral vein or the carotid artery, or by stereotactic injection into the rat pituitary. Double-label histology was used to evaluate cell-type expression in the pituitary. Intravascular injection of AdGHGal or AdalphaGal failed to deliver the marker gene to the pituitary. However, direct stereotactic injection of recombinant adenoviral vectors into the pituitary achieved a high level of transgene expression. In addition, immunohistochemical staining revealed selective expression of the AdGHGal or AdalphaGal transgenes in pituitary cells that normally produce the respective hormones. These findings indicate that adenoviral vectors carrying pituitary gland-specific promoters may be useful for targeted gene therapy of pituitary diseases. However, because of low transduction after peripheral administration, stereotactic injection or local administration of viruses at the time of pituitary surgery is probably required for efficient gene expression.


An evaluation of the Reltus ear massager for short-term tinnitus relief.
Auditory stimulation has been shown to suppress the loudness of tinnitus (residual inhibition, RI). Somatosensory manipulations have also been shown to sometimes decrease tinnitus perception. An 'ear-massaging' device, the 'Reltus' ( www.reltus.com ), has been marketed as a tinnitus treatment device. This study was undertaken to evaluate its short-term effectiveness and mode of effect. The research was undertaken in two phases. Phase 1 measured the change in tinnitus perception after one minute of auditory stimulation through headphones and after application of a vibrating device to four different stimulation points around the pinna for one minute each. Phase 2 evaluated if it was the vibrations that were responsible for the effect on tinnitus perception, or sound produced by the vibrator. Twenty-three participants completed phase 1 and 10 participants participated in the second phase. RI to auditory stimulation was found in 87% of participants and to tactile stimulation in 83%. No significant differences were found in the effectiveness between the four vibration stimulation points, or between the left and right ear of the participants. The Reltus produced a sound that resulted in RI. It is the auditory artifact of the Reltus that was responsible for short-term tinnitus suppression.


A metabolic model for members of the genus Tetrasphaera involved in enhanced biological phosphorus removal.
Members of the genus Tetrasphaera are considered to be putative polyphosphate accumulating organisms (PAOs) in enhanced biological phosphorus removal (EBPR) from wastewater. Although abundant in Danish full-scale wastewater EBPR plants, how similar their ecophysiology is to 'Candidatus Accumulibacter phosphatis' is unclear, although they may occupy different ecological niches in EBPR communities. The genomes of four Tetrasphaera isolates (T. australiensis, T. japonica, T. elongata and T. jenkinsii) were sequenced and annotated, and the data used to construct metabolic models. These models incorporate central aspects of carbon and phosphorus metabolism critical to understanding their behavior under the alternating anaerobic/aerobic conditions encountered in EBPR systems. Key features of these metabolic pathways were investigated in pure cultures, although poor growth limited their analyses to T. japonica and T. elongata. Based on the models, we propose that under anaerobic conditions the Tetrasphaera-related PAOs take up glucose and ferment this to succinate and other components. They also synthesize glycogen as a storage polymer, using energy generated from the degradation of stored polyphosphate and substrate fermentation. During the aerobic phase, the stored glycogen is catabolized to provide energy for growth and to replenish the intracellular polyphosphate reserves needed for subsequent anaerobic metabolism. They are also able to denitrify. This physiology is markedly different to that displayed by 'Candidatus Accumulibacter phosphatis', and reveals Tetrasphaera populations to be unusual and physiologically versatile PAOs carrying out denitrification, fermentation and polyphosphate accumulation.


Optimisation of window settings for traditional and noise-optimised virtual monoenergetic imaging in dual-energy computed tomography pulmonary angiography.
To define optimal window settings for displaying virtual monoenergetic images (VMI) of dual-energy CT pulmonary angiography (DE-CTPA). Forty-five patients who underwent clinically-indicated third-generation dual-source DE-CTPA were retrospectively evaluated. Standard linearly-blended (M_0.6), 70-keV traditional VMI (M70), and 40-keV noise-optimised VMI (M40+) reconstructions were analysed. For M70 and M40+ datasets, the subjectively best window setting (width and level, B-W/L) was independently determined by two observers and subsequently related with pulmonary artery attenuation to calculate separate optimised values (O-W/L) using linear regression. Subjective evaluation of image quality (IQ) between W/L settings were assessed by two additional readers. Repeated measures of variance were performed to compare W/L settings and IQ indices between M_0.6, M70, and M40+. B-W/L and O-W/L for M70 were 460/140 and 450/140, and were 1100/380 and 1070/380 for M40+, respectively, differing from standard DE-CTPA W/L settings (450/100). Highest subjective scores were observed for M40+ regarding vascular contrast, embolism demarcation, and overall IQ (all p<0.001). Application of O-W/L settings is beneficial to optimise subjective IQ of VMI reconstructions of DE-CTPA. A width slightly less than two times the pulmonary trunk attenuation and a level approximately of overall pulmonary vessel attenuation are recommended. • Application of standard window settings for VMI results in inferior image perception. • No significant differences between B-W/L and O-W/L for M70/M40+ were observed. • O-W/L for M70 were 450/140 and were 1070/380 for M40+. • Improved subjective IQ characteristics were observed for VMI displayed with O-W/L.


Cone dystrophy with "supernormal" rod ERG: psychophysical testing shows comparable rod and cone temporal sensitivity losses with no gain in rod function.
We report a psychophysical investigation of 5 observers with the retinal disorder "cone dystrophy with supernormal rod ERG," caused by mutations in the gene KCNV2 that encodes a voltage-gated potassium channel found in rod and cone photoreceptors. We compared losses for rod- and for cone-mediated vision to further investigate the disorder and to assess whether the supernormal ERG is associated with any visual benefit. L-cone, S-cone, and rod temporal acuity (critical flicker fusion frequency) were measured as a function of target irradiance; L-cone temporal contrast sensitivity was measured as a function of temporal frequency. Temporal acuity measures revealed that losses for vision mediated by rods, S-cones, and L-cones are roughly equivalent. Further, the gain in rod function implied by the supernormal ERG provides no apparent benefit to near-threshold rod-mediated visual performance. The L-cone temporal contrast sensitivity function in affected observers was similar in shape to the mean normal function but only after the mean function was compressed by halving the logarithmic sensitivities. The name of this disorder is potentially misleading because the comparable losses found across rod and cone vision suggest that the disorder is a generalized cone-rod dystrophy. Temporal acuity and temporal contrast sensitivity measures are broadly consistent with the defect in the voltage-gated potassium channel producing a nonlinear distortion of the photoreceptor response but after otherwise normal transduction processes.


The return of metabolism: biochemistry and physiology of the pentose phosphate pathway.
The pentose phosphate pathway (PPP) is a fundamental component of cellular metabolism. The PPP is important to maintain carbon homoeostasis, to provide precursors for nucleotide and amino acid biosynthesis, to provide reducing molecules for anabolism, and to defeat oxidative stress. The PPP shares reactions with the Entner-Doudoroff pathway and Calvin cycle and divides into an oxidative and non-oxidative branch. The oxidative branch is highly active in most eukaryotes and converts glucose 6-phosphate into carbon dioxide, ribulose 5-phosphate and NADPH. The latter function is critical to maintain redox balance under stress situations, when cells proliferate rapidly, in ageing, and for the 'Warburg effect' of cancer cells. The non-oxidative branch instead is virtually ubiquitous, and metabolizes the glycolytic intermediates fructose 6-phosphate and glyceraldehyde 3-phosphate as well as sedoheptulose sugars, yielding ribose 5-phosphate for the synthesis of nucleic acids and sugar phosphate precursors for the synthesis of amino acids. Whereas the oxidative PPP is considered unidirectional, the non-oxidative branch can supply glycolysis with intermediates derived from ribose 5-phosphate and vice versa, depending on the biochemical demand. These functions require dynamic regulation of the PPP pathway that is achieved through hierarchical interactions between transcriptome, proteome and metabolome. Consequently, the biochemistry and regulation of this pathway, while still unresolved in many cases, are archetypal for the dynamics of the metabolic network of the cell. In this comprehensive article we review seminal work that led to the discovery and description of the pathway that date back now for 80 years, and address recent results about genetic and metabolic mechanisms that regulate its activity. These biochemical principles are discussed in the context of PPP deficiencies causing metabolic disease and the role of this pathway in biotechnology, bacterial and parasite infections, neurons, stem cell potency and cancer metabolism.


Photoluminescence quantum efficiency of Er optical centers in GaN epilayers.
We report the quantum efficiency of photoluminescence processes of Er optical centers as well as the thermal quenching mechanism in GaN epilayers prepared by metal-organic chemical vapor deposition. High resolution infrared spectroscopy and temperature dependence measurements of photoluminescence intensity from Er ions in GaN under resonant excitation excitations were performed. Data provide a picture of the thermal quenching processes and activation energy levels. By comparing the photoluminescence from Er ions in the epilayer with a reference sample of Er-doped SiO2, we find that the fraction of Er ions that emits photon at 1.54 μm upon a resonant optical excitation is approximately 68%. This result presents a significant step in the realization of GaN:Er epilayers as an optical gain medium at 1.54 μm.


Global warming: the balance of evidence and its policy implications. A review of the current state-of-the-controversy.
Global warming and attendant climate change have been controversial for at least a decade. This is largely because of its societal implications. With the recent publication of the Third Assessment Report of the United Nations' Intergovernmental Panel on Climate Change there has been renewed interest and controversy about how certain the scientific community is of its conclusions: that humans are influencing the climate and that global temperatures will continue to rise rapidly in this century. This review attempts to update what is known and in particular what advances have been made in the past 5 years or so. It does not attempt to be comprehensive. Rather it focuses on the most controversial issues, which are actually few in number. They are: Is the surface temperature record accurate or is it biased by heat from cities, etc.?, Is that record significantly different from past warmings such as the Medieval Warming Period?, Is not the sun's increasing activity the cause of most of the warming?, Can we model climate and predict its future, or is it just too complex and chaotic?, Are there any other changes in climate other than warming, and can they be attributed to the warming? Despite continued uncertainties, the review finds affirmative answers to these questions. Of particular interest are advances that seem to explain why satellites do not see as much warming as surface instruments, how we are getting a good idea of recent paleoclimates, and why the 20th century temperature record was so complex. It makes the point that in each area new information could come to light that would change our thinking on the quantitative magnitude and timing of anthropogenic warming, but it is unlikely to alter the basic conclusions. Finally, there is a very brief discussion of the societal policy response to the scientific message, and the author comments on his 2-year email discussions with many of the world's most outspoken critics of the anthropogenic warming hypothesis.


Randomized multicenter trial of sirolimus vs prednisone as initial therapy for standard-risk acute GVHD: the BMT CTN 1501 trial.
Clinical- and biomarker-based tools may identify a lower-risk acute graft-versus-host disease (GVHD) population amenable to novel, reduced-intensity treatments. Previous data suggest sirolimus may rival standard of care prednisone. We conducted a National Heart, Lung, and Blood Institute/National Cancer Institute-funded Blood and Marrow Transplant Clinical Trials Network multicenter, open-label, randomized phase 2 trial to estimate the difference in day 28 complete response (CR)/partial response (PR) rates for sirolimus vs prednisone as initial treatment of patients with standard risk (SR) acute GVHD as defined by the Minnesota (MN) GVHD Risk Score and Ann Arbor (AA1/2) biomarker status. A total of 127 MN-SR patients were randomized (1:1), and 122 were AA1/2 (sirolimus, n = 58; prednisone, n = 64). Others were AA3 (n = 4), or AA status missing (n = 1). The day 28 CR/PR rates were similar for sirolimus 64.8% (90% confidence interval [CI], 54.1%-75.5%) vs 73% (90% CI, 63.8%-82.2%) for prednisone. The day 28 rate of CR/PR with prednisone ≤0.25 mg/kg/day was significantly higher for sirolimus than prednisone (66.7% vs 31.7%; P < .001). No differences were detected in steroid-refractory acute GVHD, disease-free survival, relapse, nonrelapse mortality, or overall survival. Sirolimus was associated with reduced steroid exposure and hyperglycemia, reduced grade 2 to 3 infections, improvement in immune suppression discontinuation and patient-reported quality of life, and increased risk for thrombotic microangiopathy. For patients with clinical- and biomarker-based SR acute GVHD, sirolimus demonstrates similar overall initial treatment efficacy as prednisone. In addition, sirolimus therapy spares steroid exposure and allied toxicity, does not compromise long-term survival outcomes, and is associated with improved patient-reported quality of life. This trial was registered at www.clinicaltrials.gov as #NCT02806947.


Estimating the burden of malaria in Senegal: Bayesian zero-inflated binomial geostatistical modeling of the MIS 2008 data.
The Research Center for Human Development in Dakar (CRDH) with the technical assistance of ICF Macro and the National Malaria Control Programme (NMCP) conducted in 2008/2009 the Senegal Malaria Indicator Survey (SMIS), the first nationally representative household survey collecting parasitological data and malaria-related indicators. In this paper, we present spatially explicit parasitaemia risk estimates and number of infected children below 5 years. Geostatistical Zero-Inflated Binomial models (ZIB) were developed to take into account the large number of zero-prevalence survey locations (70%) in the data. Bayesian variable selection methods were incorporated within a geostatistical framework in order to choose the best set of environmental and climatic covariates associated with the parasitaemia risk. Model validation confirmed that the ZIB model had a better predictive ability than the standard Binomial analogue. Markov chain Monte Carlo (MCMC) methods were used for inference. Several insecticide treated nets (ITN) coverage indicators were calculated to assess the effectiveness of interventions. After adjusting for climatic and socio-economic factors, the presence of at least one ITN per every two household members and living in urban areas reduced the odds of parasitaemia by 86% and 81% respectively. Posterior estimates of the ORs related to the wealth index show a decreasing trend with the quintiles. Infection odds appear to be increasing with age. The population-adjusted prevalence ranges from 0.12% in Thillé-Boubacar to 13.1% in Dabo. Tambacounda has the highest population-adjusted predicted prevalence (8.08%) whereas the region with the highest estimated number of infected children under the age of 5 years is Kolda (13940). The contemporary map and estimates of malaria burden identify the priority areas for future control interventions and provide baseline information for monitoring and evaluation. Zero-Inflated formulations are more appropriate in modeling sparse geostatistical survey data, expected to arise more frequently as malaria research is focused on elimination.


Overview of randomised trials of diuretics in pregnancy.
Over the past 20 years at least 11 randomised trials of the prevention with diuretics of pre-eclampsia and its sequelae have been undertaken. Nine of these were reviewed. Reliable data from the remaining two were not available. The nine reviewed had investigated a total of nearly 7000 people. Significant evidence of prevention of "pre-eclampsia" was overwhelming, even when oedema was not included as a diagnostic criterion. But as the definitions of pre-eclampsia that had been used depended heavily on increases in blood pressure this evidence may simply have reflected the well known ability of diuretics to reduce blood pressure. When the data on perinatal death were reviewed a little difference was seen in postnatal survival. The incidence of stillbirths was reduced by about one third with treatment, but, perhaps owing to small numbers (only 37 stillbirths), the difference was not significant. Thus these randomised trials failed to provide reliable evidence of either the presence or the absence of any worthwhile effects of treatment with diuretics on perinatal mortality. The implications of this for current and future trials of beta blockers and other agents in the prevention of pre-eclampsia and its sequelae are that extremely large, ultra simple randomised trials are needed, of a size sufficient to permit direct assessment of the effects of treatment not on pre-eclampsia but on perinatal mortality itself. This may require the study of tens of thousands of pregnancies.


Design and modeling of electrode networks for code-division multiplexed resistive pulse sensing in microfluidic devices.
A typical microfluidic device sorts, captures or fractionates sample constituents by exposing them to discriminating microenvironments. Direct electronic acquisition of such manipulation by a network of integrated sensors can provide a fast, integrated readout, replacing otherwise required microscopy. We have recently introduced a sensor technology, Microfluidic CODES, which allows us to multiplex resistive pulse sensors on a microfluidic device. Microfluidic CODES employs a network of micromachined coplanar electrodes such that particles passing over these electrodes produce distinguishable code sequences. In this paper, we explain the design process to specifically generate an orthogonal digital code set for an efficient and accurate demultiplexing of the sensor signals. We also introduce an equivalent circuit model for a network of code-multiplexed resistive pulse sensors by utilizing the Foster-Schwan model and conformal mapping, to model dynamic cell-electrode interaction in a non-uniform electric field. Our results closely match with both experimental measurements using cell lines and finite element analysis. The coding and modeling framework presented here will enable the design of code-division multiplexed resistive pulse sensors optimized to produce desired waveform patterns to ensure reliable and efficient decoding.


Normal weight individuals who develop type 2 diabetes: the personal fat threshold.
Type 2 diabetes (T2DM) is frequently regarded as a disease of obesity and its occurrence in individuals of normal body mass index (BMI) is often regarded as indicating a non-obesity-related subtype. However, the evidence for such a distinct, common subtype is lacking. The United Kingdom Prospective Diabetes Study (UKPDS) cohort of people diagnosed with T2DM in the 1970s and 1980s had a median BMI of only 28 kg/m2. UKPDS data form the basis of current understanding of the condition even though one in three of those studied had a BMI of less than 25 kg/m2. BMI, though, is a population measure and not a rigid personal guide. Weight loss is considered de rigueur for treating obese diabetic individuals, but it is not usually considered for those deemed to have a normal BMI. Given the new evidence that early T2DM can be reversed to normal glucose tolerance by substantial weight loss, it is important to explain why non-overweight people respond to this intervention as well as obese individuals. We hypothesize that each individual has a personal fat threshold (PFT) which, if exceeded, makes likely the development of T2DM. Subsequent weight loss to take the individual below their level of susceptibility should allow return to normal glucose control. Crucially, the hypothesized PFT is independent of BMI. It allows both understanding of development of T2DM in the non-obese and remission of diabetes after substantial weight loss in people who remain obese by definition. To illustrate this concept, we present the distribution curve of BMI at diagnosis for the UKPDS cohort, together with a diagram explaining individual behaviour within the population. The concept of PFT is of practical benefit in explaining the onset of diabetes and its logical management to the non-obese majority of people with T2DM.


MicroRNAs as new player in rheumatoid arthritis.
MicroRNAs (miRNAs) are small noncoding RNA molecules that negatively regulate gene expression at the post-transcriptional level. Currently, there are 939 mature human miRNA sequences listed in the Sanger updated miRNA registry. There are approximately 1500 predicted miRNAs in the human genome that may regulate the expression of one third of our genes. By controlling the accumulation of the target protein(s) in cells, these regulatory RNA molecules participate in key functions in many physiological networks and their deregulation has been implicated in the pathogenesis of serious human disorders, such as cancer and infection. The implication of miRNAs in immune-mediated disorders such as rheumatoid arthritis (RA) has recently emerged suggesting that miRNA-based therapeutic approaches may have a promising potential in these diseases. Here, we provide an overview of the state-of-the-art on miRNAs in RA, focusing on both systemic and local features of the pathology.


Pycnodysostosis with Special Emphasis on Dentofacial Characteristics.
Pycnodysostosis is an autosomal recessive disorder that manifests as osteosclerosis of the skeleton due to the defective osteoclasts mediated bone turnover. The diagnosis of this disorder is established on the basis of its characteristic features and must be differentially diagnosed with other bone disorders. Dental surgeons should be aware of the limitations and possible adverse oral complications such as osteomyelitis of bone in these patients. This will guide them in planning realistic treatment goals. This paper reports the clinical and radiographic features of pycnodysostosis with the great emphasis on its dentofacial characteristics. The aim of this case report is to give an insight into the etiology, pathogenesis, and differential diagnosis of this disorder and to prepare the dentists and maxillofacial surgeons to overcome the challenges in treating these patients.


Hypocapnia Alone Fails to Provoke Important Electrocardiogram Changes in Coronary Artery Diseased Patients.
There is still an urgent clinical need to develop non-invasive diagnostic tests for early ischemic heart disease because, once angina occurs, it is too late. Hypocapnia has long been known to cause coronary artery vasoconstriction. Some new cardiology tests are accompanied by the claim that they must have potential diagnostic value if hypocapnia enhances their cardiac effects in healthy subjects. But no previous study has tested whether hypocapnia produces bigger cardiac effects in patients with angina than in healthy subjects. Severe hypocapnia (a PetCO2 level of 20 mmHg) lasting >15 min was mechanically induced by facemask, while conscious and unmedicated, in 18 healthy subjects and in 10 patients with angina and angiographically confirmed coronary artery disease, awaiting by-pass surgery. Each participant was their own control in normocapnia (where CO2 was added to the inspirate) and the order of normocapnia and hypocapnia was randomized. Twelve lead electrocardiograms (ECG) were recorded and automated measurements were made on all ECG waveforms averaged over >120 beats. 2D echocardiography was also performed on healthy subjects. In the 18 healthy subjects, we confirm that severe hypocapnia (a mean PetCO2 of 20 ± 0 mmHg, P < 0.0001) consistently increased the mean T wave amplitude in leads V1-V3, but by only 31% (P < 0.01), 15% (P < 0.001) and 11% (P < 0.05), respectively. Hypocapnia produced no other significant effects (p > 0.05) on their electro- or echocardiogram. All 10 angina patients tolerated the mechanical hyperventilation well, with minimal discomfort. Hypocpania caused a similar increase in V1 (by 39%, P < 0.05 vs. baseline, but P > 0.05 vs. healthy controls) and did not induce angina. Its effects were no greater in patients who did not take β-blockers, or did not take organic nitrates, or had the worst Canadian Cardiovascular Society scores. Non-invasive mechanical hyperventilation while awake and unmedicated is safe and acceptable, even to patients with angina. Using it to produce severe and prolonged hypocapnia alone does produce significant ECG changes in angina patients. But its potential diagnostic value for identifying patients with coronary stenosis requires further evaluation.


The HyPRP gene EARLI1 has an auxiliary role for germinability and early seedling development under low temperature and salt stress conditions in Arabidopsis thaliana.
The effect of the hybrid proline-rich protein (HyPRP) gene EARLI1 on the rate of germination (germinability) of Arabidopsis seeds and seedling growth under low temperature and salt stress conditions was investigated. EARLI1 was induced during germination in embryonic tissues, and was strongly expressed in certain parts of young seedlings. Comparisons of control, overexpressing (OX), and knockout (KO) lines indicated that higher than wild type levels of EARLI1 improved germinability, root elongation, and reduction of sodium accumulation in leaves under salt stress, as well as germinability under low-temperature stress. Abscisic acid (ABA) contents were relatively low after prolonged salt stress, suggesting that EARLI1 has an ABA-independent effect on germinability under these conditions. Overexpression of EARLI1 during germination enhanced the sensitivity of seeds to exogenously applied ABA, suggesting that EARLI1 has an ABA-dependent negative effect on seed germinability under high ABA stress conditions. Well-known stress response marker genes such as COR15a, KIN1, P5SC1, and RD29 were unaffected whereas P5SC2, RD22, or RAB18 were only slightly affected in OX and KO plants. The pleiotropic effects of EARLI1 during stress and an absence of strong regulatory effects on stress marker genes suggest that this HyPRP gene has an auxiliary role for various stress protection responses in Arabidopsis.


Challenges of the Use of Sound Emergence for Setting Legal Noise Limits.
In the vast majority of legislation on environmental noise, the metric used for expressing limit values is based on sound pressure levels. But some countries have introduced sound emergence limit values where the compliance of a noise-generating activity is defined as a maximum allowable difference between the sound pressure level with and without the regulated activity operating. This paper investigates the foundations and the merits of this kind of differential noise limit values. Our review of literature indicates that there is very little evidence supporting the use of differential noise limits over absolute ones. Moreover, while sound emergence limits seem to originate from consideration about audibility of the regulated noise source, they appear to give little insight into what is audible and what is not. Furthermore, both the definition and the practical measurement of sound emergence raise several challenges that compromise reproducibility. In addition, first, the reference to background noise makes it very difficult first to ascertain the conformity of noisy installations in the long run, second to effectively protect the community from excessive noise and third to evaluate conformity on the basis of simulations. When switching to another metric is not an option the paper makes recommendations toward a more reliable use of sound emergence.


Echinococcus multilocularis in south-eastern Europe (Romania).
Echinococcus multilocularis, the causative agent of alveolar echinococcosis (AE) in humans, has been found in 4.8% of 561 red foxes originating from various regions of Romania. Infected foxes were identified in 8 of 15 counties with average prevalence rates between 1.7% and 14.6%. In previous studies, E. multilocularis was not found in 535 foxes from three counties, but larval stages (metacestodes) were present in four species of rodents. Furthermore, AE was diagnosed in two patients. Experiences from other European regions indicate that several factors (such as increasing fox populations with higher parasite prevalences and urban cycles of the parasite) may result in an increased infection risk for humans.


Laparoscopic antireflux surgery. What is real progress?
The authors aim to substantiate, with objective arguments, potential advantages of laparoscopic versus open antireflux surgery in the light of the recent crude experience of the Louvain Medical School Hospital. Seventy-two consecutive patients with disabling gastroesophageal reflux disease ([GERD], n = 56), symptomatic hiatal hernia without GERD (n = 5), or unsatisfactory outcome after unsuccessful antireflux procedure (n = 11) were operated on by laparotomy (n = 28), laparoscopy (n = 39), or thoracotomy (n = 5). The antireflux procedure was a subdiaphragmatic Nissen fundoplication (n = 60), an intrathoracic Nissen fundoplication (short esophagus, n = 3), a subdiaphragmatic 240 degrees fundoplication (severe motility disorders, n = 3), a Lortat-Jacob repair (hiatal hernia without GERD, n = 5), and a duodenal diversion (delayed gastric emptying, n = 1). Major postoperative morbidity included two pulmonary embolisms (one laparoscopy patient and one laparotomy patient), and one hemothorax (one thoracotomy patient). Mean hospital stay was 6.4 days for laparoscopy, 7.8 days for laparotomy, and 12.5 days for thoracotomy. Postoperative morphine consumption (patient-controlled analgesia) averaged 47 mg/48 hrs (laparoscopy) versus 46 mg/48 hrs (laparotomy with primary antireflux surgery) (p > 0.05). Although 93% of the laparoscopy patients returned to work within 3 weeks after surgery, 92% of the laparotomy and thoracotomy patients resumed their activity after more than 6 weeks. At follow-up, 87.5% of the patients were asymptomatic or had inconsequential symptoms, 9.8% had disabling side effects, and 2.7% had persistent or recurring esophageal symptoms. There were four parietal herniations, i.e., one incisional hernia and one recurrence of a repaired umbilical hernia in the laparotomy group, and two herniations of the wrap into the chest--probably related to a premature return to manual work--in the laparoscopy group. Three laparoscopy patients were dissatisfied with the esthetics of their scars. Lower esophageal sphincter pressure and esophageal acid exposure in the laparoscopy patients who were investigated were normal in 100% and 95%, respectively. Laparoscopy is a good approach for achieving successful antireflux surgery in selected cases. However, its fails to substantially reduce postoperative complication rate and discomfort, duration of the hospital stay, and the risk of esthetic sequela. Early return to work is questionable for manual workers. The subdiaphragmatic Nissen fundoplication is not an all-purpose antireflux procedure.


The influence of ration size on copper homeostasis during sublethal dietary copper exposure in juvenile rainbow trout, Oncorhynchus mykiss.
The influence of ration size on homeostasis and sublethal toxicity of copper (Cu) was assessed in rainbow trout (Oncorhynchus mykiss) during dietary Cu exposure in synthetic soft water. A constant dietary dose of 0.24 micromol Cu per g fish per day as CuSO(4).5H(2)O was delivered via diets containing 15.75, 7.87, and 5.24 micromol Cu g(-1) fed at 1.5, 3.0, and 4.5% wet body weight daily ration, respectively. Juvenile rainbow trout showed clear effects of ration but not Cu on growth suggesting that growth is hardly a sensitive endpoint for detection of sublethal dietary Cu exposure. All Cu-exposed fish accumulated the same total metal load when expressed on a per fish basis. This suggests that differences in tissue and whole-body Cu concentrations among the treatments reflected the differences in the fish size rather than total Cu accumulation, and demonstrate that absorption and accumulation of Cu from the gut during dietary exposure are independent of the food quantity in which the Cu is delivered. Fish fed a high ration exhibited greater mass-specific unidirectional uptake of waterborne Cu than fish fed a low ration indicating an increased need for Cu for growth processes in rapidly growing fish. Stimulated excretion of Cu was indicated by greater Cu accumulation in the bile of the Cu-exposed fish. Branchial Na(+), K(+)-ATPase was not affected by dietary Cu exposure or ration but gut Na(+), K(+)-ATPase activities showed stimulatory effects of increasing ration but not of Cu exposure. The 96-h LC50 for waterborne Cu (range 0.17-0.21 micromol l(-1) (10.52-13.20 microg l(-1)) was the same in all treatment groups indicating that ration size was unimportant and that dietary Cu did not induce an increase in tolerance to waterborne Cu. Taken together these results suggest that the nutritional status, fish size, and growth rates should be considered when comparing whole-body and tissue Cu concentration data for biomonitoring and risk assessment. Moreover, expressing the exposure as total metal dose rather than metal concentration in the diet is more appropriate.


Intermediate filament collapse is an ATP-dependent and actin-dependent process.
In this study, we have investigated the properties of intermediate filament rearrangements using experimentally induced collapse of vimentin intermediate filaments in mouse fibroblasts. In these cells, depolymerizing microtubules by colchicine or vinblastine treatment at 37 degrees C results in a two-stage collapse of intermediate filaments. First, the vimentin filaments aggregate into large cables; then, the cables coil into a dense mass surrounding the nucleus. By using inhibitors of oxidative phosphorylation along with glucose deprivation to lower intracellular ATP levels by 95%, we have found that both stages of intermediate filament collapse require ATP. However, once collapse has occurred, only the second stage can be reversed in the absence of microtubules by lowering ATP levels. An additional difference between the two stages of collapse was revealed by treating cells with cytochalasin D: the formation of intermediate filament cables still occurs after disruption of the actin filament system by cytochalasin, but the subsequent coiling of cables to form a perinuclear mass is strongly inhibited by these conditions, and can be reversed by applying cytochalasin to cells in which intermediate filaments have already undergone complete collapse. We propose that the formation of vimentin cables involves a phosphorylation event, while the coiling of cables into a perinuclear mass relies on interaction of intermediate filaments with a component of the actin cortex.


Canine vector-borne diseases in India: a review of the literature and identification of existing knowledge gaps.
Despite the combination of favourable climate for parasites and vectors, and large populations of stray dogs, information concerning the epidemiology, diagnosis and management of canine vector-borne diseases in India is limited. However, with the country's expanding economy and adaptation to western culture, higher expectations and demands are being placed on veterinary surgeons for improved knowledge of diseases and control. This review aims to provide an overview of the current state of knowledge of these diseases in India and identify existing knowledge gaps in the literature which need to be addressed. The available literature on this subject, although limited, suggests that a number of canine vector-borne diseases such as filariasis, babesiosis and ehrlichiosis are endemic throughout India, as diagnosed mostly by morphological methods. Detailed investigations of the epidemiology and zoonotic potential of these pathogens has been neglected. Further study is essential to develop a better understanding of the diversity of canine vector-borne diseases in India, and their significance for veterinary and public health.


Achromobacter Species Isolated from Cystic Fibrosis Patients Reveal Distinctly Different Biofilm Morphotypes.
Achromobacter species have attracted attention as emerging pathogens in cystic fibrosis. The clinical significance of Achromobacter infection is not yet fully elucidated; however, their intrinsic resistance to antimicrobials and ability to form biofilms renders them capable of establishing long-term chronic infections. Still, many aspects of Achromobacter biofilm formation remain uncharacterized. In this study, we characterized biofilm formation in clinical isolates of Achromobacter and investigated the effect of challenging the biofilm with antimicrobials and/or enzymes targeting the extracellular matrix. In vitro biofilm growth and subsequent visualization by confocal microscopy revealed distinctly different biofilm morphotypes: a surface-attached biofilm morphotype of small aggregates and an unattached biofilm morphotype of large suspended aggregates. Aggregates consistent with our in vitro findings were visualized in sputum samples from cystic fibrosis patients using an Achromobacter specific peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) probe, confirming the presence of Achromobacter biofilms in the CF lung. High antibiotic tolerance was associated with the biofilm phenotype, and biocidal antibiotic concentrations were up to 1000 fold higher than for planktonic cultures. Treatment with DNase or subtilisin partially dispersed the biofilm and reduced the tolerance to specific antimicrobials, paving the way for further research into using dispersal mechanisms to improve treatment strategies.


Principles of branch formation and branch patterning in Hydrozoa.
The freshwater polyp Hydra produces buds which separate from the parent. Other Hydrozoa produce branches which remain connected to the parent, thus forming a colony. Some Hydrozoa grow by means of an organ that is like a shoot apical meristem. Others display a sympodial type of growth. In this article, I propose that these different types of branches are organized by a common pattern-forming system. This system has self-organizing properties. It causes branch tip formation and is kept active in the tip when the tip finally differentiates into a hypostome of a polyp. The system does not cause structure formation directly but rather, determines a tissue property called positional value, in such a way that a gradient of values forms in the tissue of the bud or branch. The local value determines the local morphodynamic processes, including differentiation of the hypostome (highest positional value), tentacles and basal disc and of the exoskeleton pattern along the shoot. A high positional value favors the onset of a new self-organizing process and by lateral inhibition, such a process prevents the initiation of a further process in its surroundings. Small quantitative differences in the range of the signals involved determine whether a bud or a branch forms and whether monopodial and sympodial growth follows.


What do we know about brief interventions for physical activity that could be delivered in primary care consultations? A systematic review of reviews.
This systematic review of reviews aims to investigate how brief interventions (BIs) are defined, whether they increase physical activity, which factors influence their effectiveness, who they are effective for, and whether they are feasible and acceptable. We searched CINAHL, Cochrane database of systematic reviews, DARE, HTA database, EMBASE, MEDLINE, PsycINFO, Science Citation Index-Expanded and Social Sciences Citation Index, and Scottish Intercollegiate Guidelines Network from their inception until May 2015 to identify systematic reviews of the effectiveness of BIs aimed at promoting physical activity in adults, reporting a physical activity outcome and at least one BI that could be delivered in a primary care setting. A narrative synthesis was conducted. We identified three specific BI reviews and thirteen general reviews of physical activity interventions that met the inclusion criteria. The BI reviews reported varying definitions of BIs, only one of which specified a maximum duration of 30min. BIs can increase self-reported physical activity in the short term, but there is insufficient evidence about their long-term impact, their impact on objectively measured physical activity, and about the factors that influence their effectiveness, feasibility and acceptability. Current definitions include BIs that are too long for primary care consultations. Practitioners, commissioners and policy makers should be aware of this when interpreting evidence about BIs, and future research should develop and evaluate very brief interventions (of 5min or less) that could be delivered in a primary care consultation.


A new medical research model: ethically and responsibly advancing health for humans and animals.
With the increasing use of genomics, computational analytics, emerging technologies, and personalized medicine, the possibility of a new research model is emerging. Using the clues from thousands of species living on our planet, scientists from many disciplines (medicine, veterinary medicine, wildlife) must collaborate, prioritize, and strategize on how to address causes of health and disease. Such clues should guide disease prevention, as well as the development of innovative, efficacious, and gentler therapies. Geographic and language barriers must be broken down, and scientists--even within a single academic, corporate, or government research site--must be vigilant in seeking the help of nonmedical disciplines of colleagues from whence answers might come. The public will become more interested in and demanding of such a model, desiring that all family members (humans and animals) have an opportunity for a long and healthy life. Above all, such activities will be humanely conducted with outcomes having the greatest chance for success.


DNA-DNA interactions in bacteriophage capsids are responsible for the observed DNA knotting.
Recent experiments showed that the linear double-stranded DNA in bacteriophage capsids is both highly knotted and neatly structured. What is the physical basis of this organization? Here we show evidence from stochastic simulation techniques that suggests that a key element is the tendency of contacting DNA strands to order, as in cholesteric liquid crystals. This interaction favors their preferential juxtaposition at a small twist angle, thus promoting an approximately nematic (and apolar) local order. The ordering effect dramatically impacts the geometry and topology of DNA inside phages. Accounting for this local potential allows us to reproduce the main experimental data on DNA organization in phages, including the cryo-EM observations and detailed features of the spectrum of DNA knots formed inside viral capsids. The DNA knots we observe are strongly delocalized and, intriguingly, this is shown not to interfere with genome ejection out of the phage.


Epidemiological trends and risk factors for tobacco, alcohol and drug use among adolescents in Scotland, 2002-13.
This study estimates trends in prevalence, and patterns, of individual and multiple substance use between 2002 and 2013 amongst adolescents in Scotland. The study uses data from 134 387 participants of the biennial national 'Scottish Schools Adolescent Lifestyle and Substance Use Survey' on smoking, alcohol and illicit drug use. Current regular use and current heavy use of smoking, alcohol, illicit drugs and multiple substances was measured. Time trends in the prevalence of each outcome were estimated using univariate and multivariate logistic regression. Regular smoking, alcohol, illicit drug and multiple substance use declined significantly amongst adolescents in Scotland. However, multivariate analyses that focussed upon high-risk levels of these behaviours revealed an upward linear trend in heavy alcohol (OR = 1.06; 95% CI: 1.04, 1.07) and heavy illicit drug (OR = 1.04; 95% CI: 1.00, 1.08) use (P < 0.05). Non-white pupils were more likely to be involved in individual and multiple substance use than ethnically white British pupils. In comparison to pupils from the least deprived socioeconomic quintile, pupils from the most deprived quintile had increased odds of 1.41 (95% CI: 1.02, 1.97; P < 0.05) and 1.62 (95% CI: 1.14, 2.29; P < 0.05) of being regular and heavy multiple substance users, respectively. Further effort is required to tackle heavy alcohol and heavy illicit drug use amongst adolescents in Scotland. Prevention strategies should be informed by the risk profiles of substance misusers and evidence around the clinical and cost-effectiveness of preventive interventions.


Suppression of Fas/APO-1-mediated apoptosis by mitogen-activated kinase signaling.
Jurkat T cells undergo rapid apoptosis upon stimulation of the Fas/APO-1 (CD95) receptor. We examined the role of the mitogen-activated protein kinase (MAPK) cascade as a negative regulator of Fas-mediated apoptosis. To this end, we used both physiologic and artificial activators of MAPK, all of which activate MAPK by distinct routes. MAPK activity could be efficiently elevated by two T cell mitogens, the lectin PHA and an agonistic Ab to the T cell receptor complex as well as by the type 1 and 2A phosphatase inhibitor, calyculin A, and the protein kinase C-activating phorbol ester, tetradecanoyl phorbol acetate. All these treatments were effective in preventing the characteristic early and late features of Fas-mediated apoptosis, including activation of caspases. Our results indicate that the elevated MAPK activities intervene upstream of caspase activation. The degree of MAPK activation by the different stimuli used in our study corresponds well to their potency to inhibit apoptosis, indicating that MAPK activation serves as an efficient modulator of Fas-mediated apoptosis. The role of MAPK in modulation of Fas-mediated apoptosis was further corroborated by transient transfection with constitutively active MAPK kinase, resulting in complete inhibition of the Fas response, whereas transfection with a dominant negative form of MAPK kinase had no effect. Furthermore, the apoptosis inhibitory effect of the MAPK activators could be abolished by the specific MAPK kinase inhibitor PD 098059. Modulation of Fas responses by MAPK signaling may determine the persistence of an immune response and may explain the insensitivity of recently activated T cells to Fas receptor stimulation.


Histoincompatibility and maternal immunological status as determinants of fetoplacental weight and litter size in rodents.
Studies conducted upon inbred strains of mice, hamsters and rats have shown that following the interstrain matings the now familiar covert reactivity of pregnant females to the alloantigens of their conceptuses may benefit the latter in two ways; firstly, it exerts a significant influence upon placental weight, and indirectly upon the birth weight of the fetus-allogeneic placentas tending to be heavier than syngeneic placentas, and mothers specifically presensitized against alien paternal tissue antigens gestate fetuses with heavier placentas than normal females. Specifically tolerant mothers, on the other hand, produce smaller, F1 hybrid, fetoplacental (fp) units. The classic notion that the disparity between the birth weights of F1 hybrid and homozygous offspring is due to hybrid vigor has been challenged by the finding that DA and (DA times F1)F1 hybrid blastocysts transferred to the uteri of genetically tolerant (DA times F1)F1 hybrid rats produce fp units of similar weight Maternal immunological reactivity against the fetus qua allograft may make a significant contribution here. Additional support for the premise that maternal reactivity against fetal alloantigens in some way promotes the growth of the fp unit was afforded by the finding that excision of the para-aortic lymph nodes (which drain the uterine horns) from females before interstrain matings resulted in smaller fp units than in females subjected to sham operations. The finding with one rat strain combination that passive immunization of females with serum against their F1 hybrid conceptuses promoted the growth of the latter suggests that a humoral rather than a cellular immunity may be involved. Secondly, in the three species studied, it was observed that genetic disparity between a conceptus and its mother significantly improved its chances of implantation and development to term.


Varicella vaccine (VARIVAX) in healthy children and adolescents: results from clinical trials, 1987 to 1989.
A total of 3303 healthy children and adolescents, aged 12 months to 17 years, were vaccinated with one of five production lots of a live attenuated varicella vaccine (VARIVAX) containing 1000 to 1625 plaque-forming units per dose. The vaccine was generally well tolerated. Ninety-six percent (2381/2475) of vaccinees responded to vaccination by producing antibody as measured by a glycoprotein-based enzyme-linked immunosorbent assay; 99% (569/576) of those tested maintained antibody at 1 year following vaccination. The incidence of varicella following household exposure in vaccinees was approximately 12%; household contact historically results in 87% infection. Nearly all of the vaccinees who had varicella after vaccination had a clinically modified disease.


Adenosine triphosphate induces activation of caspase-3 in apoptosis of human granulosa-luteal cells.
Adenosine triphosphate (ATP) has been shown to induce programmed cell death in various systems. However, little is known about the effect of ATP on human granulosa-luteal cells (hGLCs). The present study was designed to examine the effect of ATP on the activation of the caspase signaling pathway and its role in inducing programmed cell death. Human GLCs were collected from patients undergoing in vitro fertilization programs, and then were cultured in FBS-supplemented DMEM for 3 days prior to our studies. To examine the dose-response relationship, hGLCs were treated with increasing concentrations of ATP (10 microM, 100 microM, 1 mM or 10 mM) for 24 hours. For time-course experiments, hGLCs were treated with 10 mM ATP for 6, 12, or 24 hours. Western blot analysis was performed using antibodies against the pro- and active forms of caspase-3, -9, or PARP. To quantify the induction of apoptosis, DNA fragmentation was measured using the cell death detection enzyme-linked immunosorbent assay. To examine the effect of human chorionic gonadotropin (hCG) in protecting cells from apoptosis, hGLCs were treated with 10 IU hCG in the presence of 10 mM ATP for 12 hours. It was demonstrated that ATP was capable of inducing DNA fragmentation in a dose- and time-dependent manner. Furthermore, Western blot analysis, which detected the pro- and active forms of caspase-3, or PARP, demonstrated that ATP activated the caspase-signaling pathway, leading to the proteolytic conversion of pro-caspase-3 to active caspase-3, and the subsequent cleavage of the caspase substrate PARP. Based on our observation, caspase-9 was not triggered by ATP. Interestingly, hCG attenuated the effect of ATP in activating the caspase signaling pathway. To our knowledge, this is the first demonstration of the ATP-induced activation of the caspase signaling pathway in the human ovary. These results support the notion that the caspase-signaling pathway is involved in mediating ATP actions in the human ovary.