Analysis of breastfeeding policies and practices in childcare centres in Adelaide, South Australia.
Breastfeeding policies and practices were analysed in childcare settings in the metropolitan area of Adelaide, South Australia. Childcare centres were purposively selected based on their geographical location, type and socioeconomic score of the area. Qualitative inquiry approach was employed by undertaking interviews with childcare centres' director or baby house coordinator to explore their perception towards breastfeeding practice and support within their centre. Breastfeeding related policy documents, where available, were also collected during the interviews to triangulate data. A total of 15 face-to-face interviews were conducted. Six childcare centres had a written policy specifically on breastfeeding support, although the technical issues of handling breastmilk were included in most centres' food and nutrition guidelines. Most participants believed that decision to breastfeed is the personal choice of parents, and hence saw the childcare centre's role as supporting parental choice whether it is breastfeeding or not. The provision of physical space to breastfeed and facilities to store the expressed breast milk were the most common practices in support of parents who had chosen to continue breastfeeding. Participants perceived mothers' work-related issues such as distance from the centre, time, and unsupportive workplace the most important barriers that led to early introduction of bottle feeding or breastfeeding cessation. Most childcare centres support breastfeeding in a more passive than active way. Breastfeeding promotion needs to be an integral part of childcare centres training, policy and practice if an increased rate of breastfeeding is to be achieved particularly amongst working mothers.


Modulation of cisplatin resistance in acquired-resistant nonsmall cell lung cancer cells.
To obtain cisplatin (CDDP)-resistant cells possessing the clinically induced resistance phenotype, H-460 nonsmall cell lung cancer cells (NSCLC) were pulse treated with 20, 60, 80 microM CDDP for 1 h every week, respectively. Twelve months later, three CDDP-resistant cell lines (H-460/CDDP20, H-460/CDDP60, H-460/CDDP80) were obtained that exhibit different levels of CDDP resistance (6- to 22-fold), and the possible mechanisms of resistance were studied. These resistant cells were cross-resistant to carboplatin and melphalan, but not to adriamycin or 5-fluorouracil. CDDP resistance in these cell lines appeared to be stable even after 6 months of growth in cisplatin-free medium. There was no evidence of drug accumulation differences between parental and resistant cells. Although both intracellular glutathione (GSH) content and glutathione S-transferase (GST) activity increased 1.5- to 2.5-fold in the resistant cells, only a minimal reversal of drug resistance was observed after buthionine sulfoximine (BSO) treatment, which depleted intracellular GSH levels. An enhancement of DNA repair activity was found in the resistant cell lines and played the major role in the cisplatin resistance phenotype. Using H-460/CDDP80 cells as a model, addition of a nontoxic concentration of pentoxifylline (PTX) significantly enhanced CDDP-induced cytotoxicity in a synergistic manner. Furthermore, more prominent reversal of CDDP resistance could be achieved when the resistant cells were pretreated with BSO, followed by PTX and CDDP combined treatment. This provides a rationale for combination therapy in refractory lung cancer using CDDP and two resistance modulators.


Antibodies to glutamic acid decarboxylase discriminate major types of diabetes mellitus.
Insulin-dependent diabetes mellitus (IDDM) is marked by circulating antibodies to a 64,000-M(r) islet cell antigen identified as glutamic acid decarboxylase (GAD). We describe a radioimmunoprecipitation assay with GAD isolated from pig brain. The sera tested were from 80 patients with IDDM including 26 with disease of recent onset and 54 with disease of longer duration (3-42 yr), 20 with non-insulin-dependent diabetes mellitus (NIDDM), and 55 nondiabetic subjects. Conventional assays for islet cell cytoplasmic antibodies were performed concurrently. The level of antibody in serum was expressed in units based on percentage reactivity of a standard reference serum. The frequency of antibody to GAD in IDDM was 69% in short-duration cases and 59% in long-duration cases. The latter was substantially higher than the frequency of islet cell cytoplasmic antibody. Antibodies to GAD were elevated (means +/- 3 SD) in 5% NIDDM cases and in none of the nondiabetic subjects. A simple laboratory test with a defined autoantigen has substantial implications for population screening and early diagnosis of IDDM and for better understanding of its pathogenesis.


A direct CO2 control system for ocean acidification experiments: testing effects on the coralline red algae Phymatolithon lusitanicum.
Most ocean acidification (OA) experimental systems rely on pH as an indirect way to control CO2. However, accurate pH measurements are difficult to obtain and shifts in temperature and/or salinity alter the relationship between pH and pCO2. Here we describe a system in which the target pCO2 is controlled via direct analysis of pCO2 in seawater. This direct type of control accommodates potential temperature and salinity shifts, as the target variable is directly measured instead of being estimated. Water in a header tank is permanently re-circulated through an air-water equilibrator. The equilibrated air is then routed to an infrared gas analyzer (IRGA) that measures pCO2 and conveys this value to a Proportional-Integral-Derivative (PID) controller. The controller commands a solenoid valve that opens and closes the CO2 flush that is bubbled into the header tank. This low-cost control system allows the maintenance of stabilized levels of pCO2 for extended periods of time ensuring accurate experimental conditions. This system was used to study the long term effect of OA on the coralline red algae Phymatolithon lusitanicum. We found that after 11 months of high CO2 exposure, photosynthesis increased with CO2 as opposed to respiration, which was positively affected by temperature. Results showed that this system is adequate to run long-term OA experiments and can be easily adapted to test other relevant variables simultaneously with CO2, such as temperature, irradiance and nutrients.


Quantitative Erythrocyte Omega-3 EPA Plus DHA Levels are Related to Higher Regional Cerebral Blood Flow on Brain SPECT.
The interrelationships between omega-3 fatty acids status, brain perfusion, and cognition are not well understood. To evaluate if SPECT brain imaging of cerebral perfusion and cognition varies as a function of omega-3 fatty acid levels. A random sample of 166 study participants was drawn from a psychiatric referral clinical for which erythrocyte quantification of omega-3 eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA) (the Omega-3 Index) was available. Quantitative brain SPECT was done on 128 regions based on a standard anatomical Atlas. Persons with erythrocyte EPA+DHA concentrations were dichotomized based on membership in top 50th percentile versus bottom 50th percentile categories. Two-sample t-tests were done to identify statistically significant differences in perfusion between the percentile groups. Partial correlations were modeled between EPA+DHA concentration and SPECT regions. Neurocognitive status was assessed using computerized testing (WebNeuro) and was separately correlated to cerebral perfusion on brain SPECT imaging and omega-3 EPA+DHA levels. Partial correlation analyses showed statistically significant relationships between higher omega-3 levels and cerebral perfusion were in the right parahippocampal gyrus (r = 0.20, p = 0.03), right precuneus (r = 0.20, p = 0.03), and vermis subregion 6 (p = 0.21, p = 0.03). Omega-3 Index levels separately correlated to the feeling subsection of the WebNeuro (r = 0.25, p = 0.01). Quantitative omega-3 EPA+DHA erythrocyte concentrations are independently correlated with brain perfusion on SPECT imaging and neurocognitive tests. These results have implications for the role of omega-3 fatty acids toward contributing to cognitive reserve.


Intermediate filament protein partnership in astrocytes.
Intermediate filaments are general constituents of the cytoskeleton. The function of these structures and the requirement for different types of intermediate filament proteins by individual cells are only partly understood. Here we have addressed the role of specific intermediate filament protein partnerships in the formation of intermediate filaments in astrocytes. Astrocytes may express three types of intermediate filament proteins: glial fibrillary acidic protein (GFAP), vimentin, and nestin. We used mice with targeted mutations in the GFAP or vimentin genes, or both, to study the impact of loss of either or both of these proteins on intermediate filament formation in cultured astrocytes and in normal or reactive astrocytes in vivo. We report that nestin cannot form intermediate filaments on its own, that vimentin may form intermediate filaments with either nestin or GFAP as obligatory partners, and that GFAP is the only intermediate filament protein of the three that may form filaments on its own. However, such filaments show abnormal organization. Aberrant intermediate filament formation is linked to diseases affecting epithelial, neuronal, and muscle cells. Here we present models by which the normal and pathogenic functions of intermediate filaments may be elucidated in astrocytes.


Mechanisms of Body Weight Reduction by Black Tea Polyphenols.
Obesity is one of the most common nutritional diseases worldwide. This disease causes health problems, such as dyslipidemia, hyperglycemia, hypertension and inflammation. There are drugs used to inhibit obesity. However, they have serious side effects outweighing their beneficial effects. Black tea, commonly referred to as "fermented tea", has shown a positive effect on reducing body weight in animal models. Black tea polyphenols are the major components in black tea which reduce body weight. Black tea polyphenols are more effective than green tea polyphenols. Black tea polyphenols exert a positive effect on inhibiting obesity involving in two major mechanisms: (i) inhibiting lipid and saccharide digestion, absorption and intake, thus reducing calorie intake; and (ii) promoting lipid metabolism by activating AMP-activated protein kinase to attenuate lipogenesis and enhance lipolysis, and decreasing lipid accumulation by inhibiting the differentiation and proliferation of preadipocytes; (iii) blocking the pathological processes of obesity and comorbidities of obesity by reducing oxidative stress. Epidemiological studies of the health relevance between anti-obesity and black tea polyphenols consumption remain to be further investigated.


Precision Medicine in the Transition to Dialysis and Personalized Renal Replacement Therapy.
Launched in 2016, the overarching goal of the Precision Medicine Initiative is to promote a personalized approach to disease management that takes into account an individual's unique underlying biology and genetics, lifestyle, and environment, in lieu of a one-size-fits-all model. The concept of precision medicine is pervasive across many areas of nephrology and has been particularly relevant to the care of advanced chronic kidney disease patients transitioning to end-stage kidney disease (ESKD). Given many uncertainties surrounding the optimal transition of incident ESKD patients to dialysis and transplantation, as well as the high mortality rates observed during this delicate transition period, there is a pressing urgency for implementing precision medicine in the management of this population. Although the traditional paradigm has been to commence incident hemodialysis patients on a 3 times/week treatment regimen, largely driven by adequacy targets, there has been growing recognition that alternative treatment regimens (ie, incremental hemodialysis) may be preferred among certain subpopulations when taking into consideration factors such as patients' residual kidney function, volume status fluctuations, symptoms, and preferences. In this review, we examine the origins of current practices in how dialysis is initiated among incident ESKD patients; incremental dialysis therapy as a dynamic and patient-centric approach that is tailored to patients' unique characteristics; recent data on the incremental hemodialysis regimen and outcomes; and future research directions using a precision nephrology approach to ESKD management with the potential to develop novel approaches, tools, and collaborative efforts to improve the health, well-being, and survival of this population.


Graft outcomes influenced by co-expression of Pax7 in graft and host tissue.
Cell replacement therapies offer promise in the treatment of neurotrauma and neurodegenerative disorders and have concentrated on the use of primary fetal brain tissue. However, there is a growing promise of using neural stem cells, in which case other factors may be important in their successful engraftment. We therefore investigated whether the co-expression of the major developmental transcription factor (Pax7 in this study) of donor tissue to graft site influences transplant survival and differentiation in the rat midbrain. Neural progenitor cells were prepared from either the Pax7-expressing dorsal (DM) or non-Pax7-expressing ventral mesencephalon (VM) of embryonic EGFP(+/+) rats. Cells were dissociated and grafted into the adult rat superior colliculus (SC) lesioned with quinolinic acid 3 days previously, a time shown to be associated with the up-regulation of Pax7. Grafts were then examined 4 weeks later. Our results suggest the origin of the graft tissue did not alter graft survival in the SC; however, dorsal grafts appear to have a higher incidence of neuronal survival, whereas ventral grafts have a higher incidence of astrocytic survivors.


Lumbopelvic Core Stabilization Exercise and Pain Modulation Among Individuals with Chronic Nonspecific Low Back Pain.
Lumbopelvic stabilization training (LPST) may provide therapeutic benefits on pain modulation in chronic nonspecific low back pain conditions. This study aimed to examine the effects of LPST on pain threshold and pain intensity in comparison with the passive automated cycling intervention and control intervention among patients with chronic nonspecific low back pain. A within-subject, repeated-measures, crossover randomized controlled design was conducted among 25 participants (7 males and 18 females) with chronic nonspecific low back pain. All the participants received 3 different types of experimental interventions, which included LPST, the passive automated cycling intervention, and the control intervention randomly, with 48 hours between the sessions. The pressure pain threshold (PPT), hot-cold pain threshold, and pain intensity were estimated before and after the interventions. Repeated-measures analysis of variance showed that LPST provided therapeutic effects as it improved the PPT beyond the placebo and control interventions (P < 0.01). The pain intensity under the LPST condition was significantly better than that under the passive automated cycling intervention and controlled intervention (P < 0.001). Heat pain threshold under the LPST condition also showed a significant trend of improvement beyond the control (P < 0.05), but no significant effects on cold pain threshold were evident. Lumbopelvic stabilization training may provide therapeutic effects by inducing pain modulation through an improvement in the pain threshold and reduction in pain intensity. LPST may be considered as part of the management programs for treatment of chronic low back pain.


Excitation-emission fluorimeter based on linear interference filters.
We describe the design, properties, and performance of an excitation-emission (EE) fluorimeter that enables spectral characterization of an object simultaneously with respect to both its excitation and its emission properties. Such devices require two wavelength-selecting elements, one in the optical path of the excitation broadband light to obtain tunable excitation and the other to analyze the resulting fluorescence. Existing EE instruments are usually implemented with two monochromators. The key feature of our EE fluorimeter is that it employs lightweight and compact linear interference filters (LIFs) as the wavelength-selection elements. The spectral tuning of both the excitation and the detection LIFs is achieved by their mechanical shift relative to each other by use of two computer-controlled linear step motors. The performance of the LIF-based EE fluorimeter is demonstrated with the fluorescent spectra of various dyes and their mixtures.


American Neurogastroenterology and Motility Society Task Force Recommendations for Resumption of Motility Laboratory Operations During the COVID-19 Pandemic.
The American Neurogastroenterology and Motility Society Task Force recommends that gastrointestinal motility procedures should be performed in motility laboratories adhering to the strict recommendations and personal protective equipment (PPE) measures to protect patients, ancillary staff, and motility allied health professionals. When available and within constraints of institutional guidelines, it is preferable for patients scheduled for motility procedures to complete a coronavirus disease 2019 (COVID-19) test within 48 hours before their procedure, similar to the recommendations before endoscopy made by gastroenterology societies. COVID-19 test results must be documented before performing procedures. If procedures are to be performed without a COVID-19 test, full PPE use is recommended, along with all social distancing and infection control measures. Because patients with suspected motility disorders may require multiple procedures, sequential scheduling of procedures should be considered to minimize need for repeat COVID-19 testing. The strategies for and timing of procedure(s) should be adapted, taking into consideration local institutional standards, with the provision for screening without testing in low prevalence areas. If tested positive for COVID-19, subsequent negative testing may be required before scheduling a motility procedure (timing is variable). Specific recommendations for each motility procedure including triaging, indications, PPE use, and alternatives to motility procedures are detailed in the document. These recommendations may evolve as understanding of virus transmission and prevalence of COVID-19 infection in the community changes over the upcoming months.


Detection of animals in natural images using far peripheral vision.
It is generally believed that the acuity of the peripheral visual field is too poor to allow accurate object recognition and, that to be identified, most objects need to be brought into foveal vision by using saccadic eye movements. However, most measures of form vision in the periphery have been done at eccentricities below 10 degrees and have used relatively artificial stimuli such as letters, digits and compound Gabor patterns. Little is known about how such data would apply in the case of more naturalistic stimuli. Here humans were required to categorize briefly flashed (28 ms) unmasked photographs of natural scenes (39 degrees high, and 26 degrees across) on the basis of whether or not they contained an animal. The photographs appeared randomly in nine locations across virtually the entire extent of the horizontal visual field. Accuracy was 93.3% for central vision and decreased almost linearly with increasing eccentricity (89.8% at 13 degrees, 76.1% at 44.5 degrees and 71.2% at 57.5 degrees ). Even at the most extreme eccentricity, where the images were centred at 70.5 degrees, subjects scored 60.5% correct. No evidence was found for hemispheric specialization. This level of performance was achieved despite the fact that the position of the image was unpredictable, ruling out the use of precued attention to target locations. The results demonstrate that even high-level visual tasks involving object vision can be performed using the relatively coarse information provided by the peripheral retina.


Unravelling the Complete Genome of Archangium gephyra DSM 2261T and Evolutionary Insights into Myxobacterial Chitinases.
Family Cystobacteraceae is a group of eubacteria within order Myxococcales and class Deltaproteobacteria that includes more than 20 species belonging to 6 genera, that is, Angiococcus, Archangium, Cystobacter, Hyalangium, Melittangium, and Stigmatella. Earlier these members have been classified based on chitin degrading efficiency such as Cystobacter fuscus and Stigmatella aurantiaca, which are efficient chitin degraders, C. violaceus a partial chitin degrader and Archangium gephyra a chitin nondegrader. Here we report the 12.5 Mbp complete genome of A. gephyra DSM 2261T and compare it with four available genomes within the family Cystobacteraceae. Phylogeny and DNA-DNA hybridization studies reveal that A. gephyra is closest to Angiococcus disciformis, C. violaceus and C. ferrugineus, which are partial chitin degraders of the family Cystobacteraceae. Homology studies reveal the conservation of approximately half of the proteins in these genomes, with about 15% unique proteins in each genome. The total carbohydrate-active enzymes (CAZome) analysis reveals the presence of one GH18 chitinase in the A. gephyra genome whereas eight copies are present in C. fuscus and S. aurantiaca. Evolutionary studies of myxobacterial GH18 chitinases reveal that most of them are likely related to Terrabacteria and Proteobacteria whereas the Archangium GH18 homolog shares maximum similarity with those of chitin nondegrading Acidobacteria.


The majority of human immunodeficiency virus type 1 particles present within splenic germinal centres are produced locally.
In most stages of human immunodeficiency virus (HIV) infection, cell-free viral particles can be detected in germinal centres (GCs) that are principally retained, in the form of immune complexes, on the surface of follicular dendritic cells (FDCs). The source of this virus remains unknown, although it is agreed that the FDCs themselves are not infected productively. By sequencing HIV viral DNA, genomic RNA and spliced mRNA isolated from individual splenic white pulps, it was shown here that the majority of HIV-1 viral particles are produced locally within the supporting lymphoid structure and do not result from trapping of circulating viruses or immune complexes. These findings underline the exquisite spatial organization of HIV-1 replication in vivo, suggesting a local origin for viruses trapped in splenic GCs.


Systematic review and meta-analysis of metacestodes prevalence in small ruminants in Ethiopia.
Metacestodes, the larval stages of canid cestode parasites, are among the causes of morbidity, mortality and financial losses in small ruminants in Ethiopia as a result of organ and carcass condemnation at slaughter. Several studies have been conducted over the years; however, these studies often had limited scope and coverage. This systematic review and meta-analysis was conducted to collate the information so far available in order to provide a pooled prevalence estimate at national level and identify potential predictors. Published and grey literature written in English and Amharic in the period from, 1st of January 1990 to June 25, 2015 were searched from electronic databases and repositories of academic and research institutions. Relevant animal level data on 67,743 small ruminants was extracted from 23 published articles and one master's thesis resulting altogether in 86 animal level reports that conformed to predefined criteria. The dataset was analyzed using a meta-analytical approach. The pooled prevalence estimate computed for metacestodes infection was 11.8% with a 95% confidence interval (CI) of 9.1, 15.4. The highest prevalence estimate 31.2% (95% CI: 23.1, 40.9) was found for Cysticercus tenuicollis (Taenia hydatigena) followed by cystic echinococcosis (Echinococcus granulosus) 8.8% (95% CI: 5.8, 13.1), Cysticercus ovis (Taenia ovis) 4.9% (95% CI: 2.9, 9.4) and Coenurus cerebralis (Taenia multiceps) 4.6% (95% CI: 1.6, 12.2). Among the predictors considered for heterogeneity analysis only sample size and metacestode type fitted the final multivariable meta-regression model and explained 26.3% of the explainable heterogeneity between studies (p<0.05). The prevalence was noted to decrease with increasing sample size. No significant difference in prevalence was observed between sheep and goats (p>0.05). In conclusion, this review showed a widespread occurrence of metacestodes in small ruminants in Ethiopia. Thus, a holistic approach to break the life cycle of these parasitic stages is suggested, including regulatory interventions that encourage dog owners to keep their dogs confined and prevent backyard slaughter and proper management of abattoir waste disposal.


Features of the secondary structure of a protein molecule from powder diffraction data.
Protein powder diffraction is shown to be suitable for obtaining de novo solutions to the phase problem at low resolution via phasing methods such as the isomorphous replacement method. Two heavy-atom derivatives (a gadolinium derivative and a holmium derivative) of the tetragonal form of hen egg-white lysozyme were crystallized at room temperature. Using synchrotron radiation, high-quality powder patterns were collected in which pH-induced anisotropic lattice-parameter changes were exploited in order to reduce the challenging and powder-specific problem of overlapping reflections. The phasing power of two heavy-atom derivatives in a multiple isomorphous replacement analysis enabled molecular structural information to be obtained up to approximately 5.3 A resolution. At such a resolution, features of the secondary structure of the lysozyme molecule can be accurately located using programs dedicated to that effect. In addition, the quoted resolution is sufficient to determine the correct hand of the heavy-atom substructure which leads to an electron-density map representing the protein molecule of proper chirality.


Multisubstance use as a feature of addiction to anabolic-androgenic steroids.
The aim of this study was to explore and describe total drug use among anabolic-androgenic steroid (AAS) users and the reasons given for the use of these drugs. The study was based on semi-structured interviews and questionnaires involving 32 patients who were attending an addiction centre in Orebro, Sweden, for AAS use. The results indicated that a history of polysubstance use among the patients was frequent. Over half were using drugs of abuse and also taking various other pharmaceuticals. Almost half of the patients took human growth hormones, and almost half of the interviewed persons were drinking alcohol to a hazardous or harmful extent. The most common reason given for taking AAS and other hormones was to increase muscle mass and strength, but some participants also used insulin as a means of losing fat. Cannabis was used to improve sleep, heroin to decrease pain and amphetamine to increase endurance and burn fat. Our data suggest that most of the current AAS users who have been admitted to a treatment programme are multiple drug users with polysubstance dependence. The study stresses the importance of carefully examining total drug use as part of the assessment regimen for this group.


Supervised detection of conserved motifs in DNA sequences with cosmo.
A number of computational methods have been proposed for identifying transcription factor binding sites from a set of unaligned sequences that are thought to share the motif in question. We here introduce an algorithm, called cosmo, that allows this search to be supervised by specifying a set of constraints that the position weight matrix of the unknown motif must satisfy. Such constraints may be formulated, for example, on the basis of prior knowledge about the structure of the transcription factor in question. The algorithm is based on the same two-component multinomial mixture model used by MEME, with stronger reliance, however, on the likelihood principle instead of more ad-hoc criteria like the E-value. The intensity parameter in the ZOOPS and TCM models, for instance, is estimated based on a profile-likelihood approach, and the width of the unknown motif is selected based on BIC. These changes allow cosmo to outperform MEME even in the absence of any constraints, as evidenced by 2- to 3-fold greater sensitivity in some simulation studies. Additional improvements in performance can be achieved by selecting the model type (OOPS, ZOOPS, or TCM) data-adaptively or by supplying correctly specified constraints, especially if the motif appears only as a weak signal in the data. The algorithm can data-adaptively choose between working in a given constrained model or in the completely unconstrained model, guarding against the risk of supplying mis-specified constraints. Simulation studies suggest that this approach can offer 3 to 3.5 times greater sensitivity than MEME. The algorithm has been implemented in the form of a stand-alone C program as well as a web application that can be accessed at http://cosmoweb.berkeley.edu. An R package is available through Bioconductor (http://bioconductor.org).


Access to information on home- and community-based services and functional status.
To examine differences in access to home- and community-based service (HCBS) information among older adults of different functional status. Cross-sectional survey of 5435 out of 11,000 randomly selected Swiss older community dwellers from the state of Vaud. Analyses stratified by functional status examined characteristics associated with limited access to HCBS information, based on self-reported knowledge on where to find information on 13 HCBS. Proportionally, more individuals did not know where to look for information in the functionally vulnerable group than in the robust and dependent ones for virtually each service. Among robust individuals, males and persons with low financial status had increased odds of limited access. Low financial status was also negatively associated with access to information among vulnerable people. Belonging to the youngest group increased the odds of limited access for dependent individuals. Efforts should be made to improve elders' access to HCBS information by developing specific strategies relevant to each functional status group. Further studies on access to HCBS information should be conducted using complex conceptual frameworks as it has been done for HCBS use.


Oncolytic activity of p53-expressing conditionally replicative adenovirus AdDelta24-p53 against human malignant glioma.
Prognosis of malignant glioma is poor, and results of treatment remain mediocre. Conditionally replicative adenoviruses hold promise as alternative anticancer agents for the treatment of malignant glioma. Here, we evaluated the conditionally replicative adenovirus AdDelta24 and its recently developed derivative AdDelta24-p53, which expresses functional p53 tumor suppressor protein while replicating in cancer cells, for treatment of malignant glioma. In comparison to its parent AdDelta24, AdDelta24-p53 killed most malignant glioma cell lines and primary glioblastoma multiforme short-term cultures more effectively, irrespective of their p53 status. Moreover, AdDelta24-p53 caused more frequent regression and more delayed growth of IGRG121 xenografts derived from a glioblastoma multiforme in vivo. Five intratumoral injections of 10(7) pfu AdDelta24 gave 24 days median tumor growth delay (P < 0.01), 30% tumor regressions, and 30% animals surviving >120 days tumor-free or with a minimal tumor residual. The same dose of AdDelta24-p53 caused >113 days of median tumor growth delay (P < 0.001), 70% tumor regressions, and 60% animals surviving >120 days tumor-free or with a minimal tumor residual. Antitumor effects in vivo were associated with extensive conditionally replicative adenovirus replication, apoptosis induction, and tumor morphology changes, including dissociation, inflammatory cell infiltration, and necrosis. We conclude that conditionally replicative adenoviruses expressing p53 are promising new agents for treatment of malignant glioma.


Nutritional manipulation of adult female Orius pumilio (Hemiptera: Anthocoridae) enhances initial predatory performance.
Commercial shipments of Orius spp. (Hemiptera: Anthocoridae) commonly include water and protein, the latter typically supplied by eggs from a moth such as Ephestia kuehniella (Zeller). To determine whether modified dietary conditions might improve predation, newly eclosed adult female Orius pumilio (Champion) were fed on E. kuehniella eggs plus encapsulated water, encapsulated 5% sucrose solution only, or encapsulated water only for periods of 24, 48, or 72 h. Feeding activity was assessed by measuring the area of a crop in digital images of dissected digestive tracts. Adult females fed continuously on eggs had larger crops than did females fed on encapsulated sucrose solution. When females were prefed encapsulated water or sucrose and then fed 3 h on eggs, their crops became highly engorged and were larger than those in females fed continuously on eggs for the same periods. In behavioral choice tests, adult females prefed on encapsulated water or 5% sucrose solution spent a larger portion of time in contact with eggs, presumably feeding, whereas females prefed on eggs showed no preference between eggs or encapsulated water. After overnight shipping, females prefed on water or sucrose and held for 48 h total consumed 3.6- and 4.3-fold, respectively, more western flower thrips, Frankliniella occidentalis (Pergande), in 3 h than those prefed on eggs. Survival rates when prefed on sugar or water were comparable with prefeeding on eggs. Thus, inundative releases of Orius can be enhanced by starvation, because females initially feed much more voraciously after shipment with no apparent reduction in fitness.


Sex and race and/or ethnicity differences in patients undergoing radiofrequency ablation for Barrett's esophagus: results from the U.S. RFA Registry.
Little is known about differences in Barrett's esophagus (BE) characteristics by sex and race and/or ethnicity or these differences in response to radiofrequency ablation (RFA). We compared disease-specific characteristics, treatment efficacy, and safety outcomes by sex and race and/or ethnicity in patients treated with RFA for BE. The U.S. RFA patient registry is a multicenter collaboration reporting processes and outcomes of care for patients treated with RFA for BE. Patients enrolled with BE. RFA. We assessed safety (stricture, bleeding, perforation, hospitalization), efficacy (complete eradication of intestinal metaplasia [CEIM]), complete eradication of dysplasia, and number of treatments to CEIM by sex and race and/or ethnicity. Among 5521 patients (4052 men; 5126 white, 137 Hispanic, 82 African American, 40 Asian, 136 heritage not identified), women were younger (60.0 vs 62.1 years) and had shorter BE segments (3.2 vs 4.4 cm) and less dysplasia (37% vs 57%) than did men. Women were almost twice as likely to stricture (odds ratio 1.7; 95% confidence interval, 1.2-2.3). Although white patients were predominantly male, about half of African Americans and Asians with BE were female. African Americans and Asians had less dysplasia than white patients. Asians and African Americans had more strictures than did white patients. There were no sex or race differences in efficacy. Observational study with non-mandated paradigms, no central laboratory for reinterpretation of pathology. In the U.S. RFA patient registry, women had shorter BE segments and less-aggressive histology. The usual tendency toward BE in men was absent in African Americans and Asians. Posttreatment stricture was more common among women and Asians. RFA efficacy did not differ by sex or race.


Multiparameter persistent homology landscapes identify immune cell spatial patterns in tumors.
Highly resolved spatial data of complex systems encode rich and nonlinear information. Quantification of heterogeneous and noisy data-often with outliers, artifacts, and mislabeled points-such as those from tissues, remains a challenge. The mathematical field that extracts information from the shape of data, topological data analysis (TDA), has expanded its capability for analyzing real-world datasets in recent years by extending theory, statistics, and computation. An extension to the standard theory to handle heterogeneous data is multiparameter persistent homology (MPH). Here we provide an application of MPH landscapes, a statistical tool with theoretical underpinnings. MPH landscapes, computed for (noisy) data from agent-based model simulations of immune cells infiltrating into a spheroid, are shown to surpass existing spatial statistics and one-parameter persistent homology. We then apply MPH landscapes to study immune cell location in digital histology images from head and neck cancer. We quantify intratumoral immune cells and find that infiltrating regulatory T cells have more prominent voids in their spatial patterns than macrophages. Finally, we consider how TDA can integrate and interrogate data of different types and scales, e.g., immune cell locations and regions with differing levels of oxygenation. This work highlights the power of MPH landscapes for quantifying, characterizing, and comparing features within the tumor microenvironment in synthetic and real datasets.


Ethical issues that confront nurses in private hospitals in the Western Cape Metropolitan area.
Nurses are faced daily with a variety of ethical issues which could be as a result of budget cuts, target setting, the shortage of nurses and expertise. The objectives of the study were to identify ethical issues related to patient care, to describe ethical issues related to patient diversity, rights and human dignity. To describe ethical issues related to caring in nursing and to the workplace environment. A quantitative explorative descriptive research design was applied. A stratified sample of (n = 142/5%) was drawn from all nurses and caregivers (N = 2990) working in a selected group of eight private hospitals. Self-administered questionnaires were used to collect the data. Statistical tests were applied to determine statistical relationships between variables. RESULTS included (95%) of respondents provided safe and committed care to their patients, (99%) loved to care for their patients and (93%) believed in the Nurses' Pledge of Service. Fifty percent (50%) of the respondents indicated verbal abuse from patients and only (59%) experienced openness and transparency in the work environment. Analysis further identified that the caregivers did not respect the noble tradition of the profession and experienced the most verbal abuse. This study has identified ethical issues which may give rise to conflict within the workplace environment if not adequately addressed by management. The study further showed that the use of caregivers not regulated in nursing practice may pose as a threat to the safety of the patient.


Elliptic flow and nuclear modification factor in ultrarelativistic heavy-ion collisions within a partonic transport model.
The quark gluon plasma produced in ultrarelativistic heavy-ion collisions exhibits remarkable features. It behaves like a nearly perfect liquid with a small shear viscosity to entropy density ratio and leads to the quenching of highly energetic particles. We show that both effects can be understood for the first time within one common framework. Employing the parton cascade Boltzmann approach to multiparton scatterings, the microscopic interactions and the space-time evolution of the quark gluon plasma are calculated by solving the relativistic Boltzmann equation. Based on cross sections obtained from perturbative QCD with explicitly taking the running coupling into account, we calculate the nuclear modification factor and elliptic flow in ultrarelativistic heavy-ion collisions. With only one single parameter associated with coherence effects of medium-induced gluon radiation, the experimental data of both observables can be understood on a microscopic level. Furthermore, we show that perturbative QCD interactions with a running coupling lead to a sufficiently small shear viscosity to entropy density ratio of the quark gluon plasma, which provides a microscopic explanation for the observations stated by hydrodynamic calculations.


Metabolic control and regulation of the tricarboxylic acid cycle in photosynthetic and heterotrophic plant tissues.
The tricarboxylic acid (TCA) cycle is a crucial component of respiratory metabolism in both photosynthetic and heterotrophic plant organs. All of the major genes of the tomato TCA cycle have been cloned recently, allowing the generation of a suite of transgenic plants in which the majority of the enzymes in the pathway are progressively decreased. Investigations of these plants have provided an almost complete view of the distribution of control in this important pathway. Our studies suggest that citrate synthase, aconitase, isocitrate dehydrogenase, succinyl CoA ligase, succinate dehydrogenase, fumarase and malate dehydrogenase have control coefficients flux for respiration of -0.4, 0.964, -0.123, 0.0008, 0.289, 0.601 and 1.76, respectively; while 2-oxoglutarate dehydrogenase is estimated to have a control coefficient of 0.786 in potato tubers. These results thus indicate that the control of this pathway is distributed among malate dehydrogenase, aconitase, fumarase, succinate dehydrogenase and 2-oxoglutarate dehydrogenase. The unusual distribution of control estimated here is consistent with specific non-cyclic flux mode and cytosolic bypasses that operate in illuminated leaves. These observations are discussed in the context of known regulatory properties of the enzymes and some illustrative examples of how the pathway responds to environmental change are given.


Fluorescence lifetime cross correlation spectroscopy resolves EGFR and antagonist interaction in live cells.
Fluorescence correlation or cross-correlation spectroscopy (FCS or FCCS), a single molecule technique, has the ability to provide highly sensitive information on interaction and dynamics of biomolecules both in vitro and in vivo. However, the inherent drawback of FCS is that species with similar molecular weight could not be differentiated. Although FCCS could resolve this through cross-correlation, it suffers from nonideal confocal volume overlap and spectral cross-talk which limits its application. In this work, we demonstrate for the first time the applicability of fluorescence lifetime correlation spectroscopy (FLCS) to monitor the interaction of an antagonist antibody with the epidermal growth factor receptor (EGFR) in live cells. As a proof of concept, we demonstrate the interaction of Cy5 labeled IgG and Alexa633 labeled anti-IgG using a single laser source (636 nm excitation) in vitro. The autocorrelation functions were separated based on their respective lifetime with a single detector and their K(d) value was determined to be 11 +/- 3 nM. An in vivo application constituting the interaction of EGFR neutralizing antibody labeled with Alexa488 and EGFR-GFP in live HEK293 cells was successfully demonstrated. The binding specificity of EGFR neutralizing antibody was confirmed by fluorescence lifetime cross-correlation measurements and fluorescence lifetime imaging (FLIM). The dissociation constant of this complex was found to be 9.2 +/- 2.7 nM. A quantitative assessment of receptor density calculations show that the density of EGFR significantly decreased, from 540 +/- 64 receptors/microm(2) to 38 +/- 7 receptors/microm(2) upon addition of the neutralizing EGFR antibody, indicating that the antagonist could induce receptor internalization. The demonstrated work not only opens up new opportunities in studying protein-protein interactions in solutions and in live cells but also provide new insights in biology to understand how the antagonists influence EGFR through live cell quantification and imaging.


Mesenchymal stem cells: a perspective from in vitro cultures to in vivo migration and niches.
Mesenchymal Stromal Progenitor/Stem Cells (MSCs) are a rare population of non-hematopoietic stromal cells, present in the bone marrow and most connective tissues of the body. They are capable of differentiation into mesenchymal tissues such as bone, cartilage, adipose tissue and muscle. In the absence of specific markers, MSCs have been defined following isolation and culture expansion, by their expression of various molecules including CD90, CD105 and CD73 and absence of markers like CD34, CD45, and CD14. MSCs have extensive proliferative ability in culture in an uncommitted state while retaining their multilineage differentiation potential, which make them attractive candidates for biological cell-based tissue repair approaches. However, their identity in their tissues of origin is not clear and the niches in which they reside are not defined. This review addresses the current state of MSC research including the differentiation potency of culture expanded MSCs, expression of chemokines and their receptors in MSCs--both relevant issues for the advocated use of MSCs for tissue repair and their systemic delivery to the affected tissues. It also reviews current knowledge of MSC niches in their native tissues, addressing the relationship with pericytes. Finally, it provides a scientific basis for the requirement of a thorough characterisation of the endogenous MSC niches within their native tissues in vivo. The knowledge of MSC niches will instruct development of innovative therapeutic measures such as producing pharmacological substances that target endogenous MSCs and their niches in order to activate and guide intrinsic repair and to improve disease outcomes.


This is Africa: an introduction to medical operations on the African continent.
This article regarding Special Operations Forces (SOF) medical operations in Africa is an introduction to a follow-on series of articles that will address in more detail pertinent medical topics which pertain to operations on the African continent. Medical operations in Africa require dynamic and systematic approaches that consider the myriad challenges, yet offer flexible solutions applied as situations and environments dictate. We believe this series of articles will be of high interest to readers and provide key information that will be germane to future SOF operations.


Fluid intake and all-cause mortality, cardiovascular mortality and kidney function: a population-based longitudinal cohort study.
Drinking eight glasses of fluid or water each day is widely believed to improve health, but evidence is sparse and conflicting. We aimed to investigate the association between fluid consumption and long-term mortality and kidney function. We conducted a longitudinal analysis within a prospective, population-based cohort study of 3858 men and women aged 49 years or older residing in Australia. Daily fluid intake from food and beverages not including water was measured using a food frequency questionnaire. We did multivariable adjusted Cox proportional hazard models for all-cause and cardiovascular mortality and a boot-strapping procedure for estimated glomerular filtration rate (eGFR). Upper and lower quartiles of daily fluid intake corresponded to >3 L and <2 L, respectively. During a median follow-up of 13.1 years (total 43 093 years at risk), 1127 deaths (26.1 per 1000 years at risk) including 580 cardiovascular deaths (13.5 per 1000 years at risk) occurred. Daily fluid intake (per 250 mL increase) was not associated with all-cause [adjusted hazard ratio (HR) 0.99 (95% CI 0.98-1.01)] or cardiovascular mortality [HR 0.98 (95% CI 0.95-1.01)]. Overall, eGFR reduced by 2.2 mL/min per 1.73 m(2) (SD 10.9) in the 1207 (31%) participants who had repeat creatinine measurements and this was not associated with fluid intake [adjusted regression coefficient 0.06 mL/min/1.73 m(2) per 250 mL increase (95% CI -0.03 to 0.14)]. Fluid intake from food and beverages excluding water is not associated with improved kidney function or reduced mortality.


Profoundly Reduced CD1c+ Myeloid Dendritic Cell HLA-DR and CD86 Expression and Increased Tumor Necrosis Factor Production in Experimental Human Blood-Stage Malaria Infection.
Dendritic cells (DCs) are sentinels of the immune system that uniquely prime naive cells and initiate adaptive immune responses. CD1c (BDCA-1) myeloid DCs (CD1c(+) mDCs) highly express HLA-DR, have a broad Toll-like receptor (TLR) repertoire, and secrete immune modulatory cytokines. To better understand immune responses to malaria, CD1c(+) mDC maturation and cytokine production were examined in healthy volunteers before and after experimental intravenous Plasmodium falciparum infection with 150- or 1,800-parasite-infected red blood cells (pRBCs). After either dose, CD1c(+) mDCs significantly reduced HLA-DR expression in prepatent infections. Circulating CD1c(+) mDCs did not upregulate HLA-DR after pRBC or TLR ligand stimulation and exhibited reduced CD86 expression. At peak parasitemia, CD1c(+) mDCs produced significantly more tumor necrosis factor (TNF), whereas interleukin-12 (IL-12) production was unchanged. Interestingly, only the 1,800-pRBC dose caused a reduction in the circulating CD1c(+) mDC count with evidence of apoptosis. The 1,800-pRBC dose produced no change in T cell IFN-γ or IL-2 production at peak parasitemia or at 3 weeks posttreatment. Overall, CD1c(+) mDCs are compromised by P. falciparum exposure, with impaired HLA-DR and CD86 expression, and have an increased capacity for TNF but not IL-12 production. A first prepatent P. falciparum infection is sufficient to modulate CD1c(+) mDC responsiveness, likely contributing to hampered effector T cell cytokine responses and assisting parasite immune evasion.


Diversity and plasticity in Rab GTPase nucleotide release mechanism has consequences for Rab activation and inactivation.
Ras superfamily GTPase activation and inactivation occur by canonical nucleotide exchange and GTP hydrolysis mechanisms. Despite conservation of active-site residues, the Ras-related Rab GTPase activation pathway differs from Ras and between different Rabs. Analysis of DENND1-Rab35, Rabex-Rab5, TRAPP-Rab1 and DrrA-Rab1 suggests Rabs have the potential for activation by distinct GDP-release pathways. Conserved active-site residues in the Rab switch II region stabilising the nucleotide-free form differentiate these pathways. For DENND1-Rab35 and DrrA-Rab1 the Rab active-site glutamine, often mutated to create constitutively active forms, is involved in GEF mediated GDP-release. By contrast, in Rab5 the switch II aspartate is required for Rabex mediated GDP-release. Furthermore, Rab1 switch II glutamine mutants refractory to activation by DrrA can be activated by TRAPP, showing that a single Rab can be activated by more than one mechanistically distinct GDP-release pathway. These findings highlight plasticity in the activation mechanisms of closely related Rab GTPases. DOI: http://dx.doi.org/10.7554/eLife.01623.001.


Smorgasbord or symphony? Assessing public health nutrition policies across 30 European countries using a novel framework.
Countries across Europe have introduced a wide variety of policies to improve nutrition. However, the sheer diversity of interventions represents a potentially bewildering smorgasbord. We aimed to map existing public health nutrition policies, and examine their perceived effectiveness, in order to inform future evidence-based diet strategies. We created a public health nutrition policy database for 30 European countries. National nutrition policies were classified and assigned using the marketing "4 Ps" approach Product (reformulation, elimination, new healthier products); Price (taxes, subsidies); Promotion (advertising, food labelling, health education) and Place (schools, workplaces, etc.). We interviewed 71 senior policy-makers, public health nutrition policy experts and academics from 14 of the 30 countries, eliciting their views on diverse current and possible nutrition strategies. Product Voluntary reformulation of foods is widespread but has variable and often modest impact. Twelve countries regulate maximum salt content in specific foods. Denmark, Austria, Iceland and Switzerland have effective trans fats bans. Price EU School Fruit Scheme subsidies are almost universal, but with variable implementation.Taxes are uncommon. However, Finland, France, Hungary and Latvia have implemented 'sugar taxes' on sugary foods and sugar-sweetened beverages. Finland, Hungary and Portugal also tax salty products. Promotion Dialogue, recommendations, nutrition guidelines, labelling, information and education campaigns are widespread. Restrictions on marketing to children are widespread but mostly voluntary. Place Interventions reducing the availability of unhealthy foods were most commonly found in schools and workplace canteens. Interviewees generally considered mandatory reformulation more effective than voluntary, and regulation and fiscal interventions much more effective than information strategies, but also politically more challenging. Public health nutrition policies in Europe appear diverse, dynamic, complex and bewildering. The "4 Ps" framework potentially offers a structured and comprehensive categorisation. Encouragingly, the majority of European countries are engaged in activities intended to increase consumption of healthy food and decrease the intake of "junk" food and sugary drinks. Leading countries include Finland, Norway, Iceland, Denmark, Hungary, Portugal and perhaps the UK. However, all countries fall short of optimal activities. More needs to be done across Europe to implement the most potentially powerful fiscal and regulatory nutrition policies.


Recognition of single-stranded RNA viruses by Toll-like receptor 7.
Viral infection of mammalian host results in the activation of innate immune responses. Toll-like receptors (TLRs) have been shown to mediate the recognition of many types of pathogens, including viruses. The genomes of viruses possess unique characteristics that are not found in mammalian genomes, such as high CpG content and double-stranded RNA. These genomic nucleic acids serve as molecular signatures associated with viral infections. Here we show that TLR7 recognizes the single-stranded RNA viruses, vesicular stomatitis virus and influenza virus. The recognition of these viruses by plasmacytoid dendritic cells and B cells through TLR7 results in their activation of costimulatory molecules and production of cytokines. Moreover, this recognition required intact endocytic pathways. Mice deficient in either the TLR7 or the TLR adaptor protein MyD88 demonstrated reduced responses to in vivo infection with vesicular stomatitis virus. These results demonstrate microbial ligand recognition by TLR7 and provide insights into the pathways used by the innate immune cells in the recognition of viral pathogens.


Supercontinuum generation in dispersion-managed tapered-rib waveguide.
We have designed a tapered-rib waveguide and numerically studied the generation of supercontinuum using such waveguides. The Air-SF57 glass-SiO(2) waveguide is 3 cm long, with a varying etched depth to manage the total dispersion. Numerical simulations are conducted for input pulses at a wavelength of 1.55 μm with a width of 150 fs and peak power of 5 kW. The proposed waveguide geometry greatly broadens the output spectrum, extending from ∼1 to ∼6 μm, caused by the continuous modification of the phase-matching condition for the generated waves.


Systematic Nanoscale Analysis of Endocytosis Links Efficient Vesicle Formation to Patterned Actin Nucleation.
Clathrin-mediated endocytosis is an essential cellular function in all eukaryotes that is driven by a self-assembled macromolecular machine of over 50 different proteins in tens to hundreds of copies. How these proteins are organized to produce endocytic vesicles with high precision and efficiency is not understood. Here, we developed high-throughput superresolution microscopy to reconstruct the nanoscale structural organization of 23 endocytic proteins from over 100,000 endocytic sites in yeast. We found that proteins assemble by radially ordered recruitment according to function. WASP family proteins form a circular nanoscale template on the membrane to spatially control actin nucleation during vesicle formation. Mathematical modeling of actin polymerization showed that this WASP nano-template optimizes force generation for membrane invagination and substantially increases the efficiency of endocytosis. Such nanoscale pre-patterning of actin nucleation may represent a general design principle for directional force generation in membrane remodeling processes such as during cell migration and division.


Anti-oxidant and anti-inflammatory activities of Inonotus obliquus and germinated brown rice extracts.
Inonotus obliquus (IO) is parasitic mushroom that grows on birch and other trees in Russia, Korea, Europe and United States. However, IO is not readily available for consumption due to its high cost and difficult growth. In this regard, IO was inoculated on germinated brown rice (GBR) in the present study and the antioxidant and anti-inflammatory activities of the IO grown on germinated brown rice (IOGBR) extracts were evaluated extensively and compared with those for IO and GBR. IOGBR showed highest antioxidant activities with scavenging total intracellular ROS and MDA levels as well as increasing the antioxidant enzymes activity in the H₂O₂-stimulated mice liver. It also exhibited best inflammatory activities by suppressing the proinflammatory mediators such as NO, PGE₂, iNOS, COX-2, TNF-α, IL-1β, and IL-6 in an LPS-stimulated RAW 264.7 cell line. This study provides a comparative approach to find out an excellent natural source of antioxidants and anti-inflammatory agent as a dietary supplement.


Social anxiety disorder and alcohol use disorder co-morbidity in the National Epidemiologic Survey on Alcohol and Related Conditions.
To assess the prevalence and clinical impact of co-morbid social anxiety disorder (SAD) and alcohol use disorders (AUD, i.e. alcohol abuse and alcohol dependence) in a nationally representative sample of adults in the United States. Data came from a large representative sample of the US population. Face-to-face interviews of 43093 adults residing in households were conducted during 2001-2002. Diagnoses of mood, anxiety, alcohol and drug use disorders and personality disorders were based on the Alcohol Use Disorder and Associated Disabilities Interview Schedule - DSM-IV version. Lifetime prevalence of co-morbid AUD and SAD in the general population was 2.4%. SAD was associated with significantly increased rates of alcohol dependence [odds ratio (OR) 2.8] and alcohol abuse (OR 1.2). Among respondents with alcohol dependence, SAD was associated with significantly more mood, anxiety, psychotic and personality disorders. Among respondents with SAD, alcohol dependence and abuse were most strongly associated with more substance use disorders, pathological gambling and antisocial personality disorders. SAD occurred before alcohol dependence in 79.7% of co-morbid cases, but co-morbidity status did not influence age of onset for either disorder. Co-morbid SAD was associated with increased severity of alcohol dependence and abuse. Respondents with co-morbid SAD and alcohol dependence or abuse reported low rates of treatment-seeking. Co-morbid lifetime AUD and SAD is a prevalent dual diagnosis, associated with substantial rates of additional co-morbidity, but remaining largely untreated. Future research should clarify the etiology of this co-morbid presentation to better identify effective means of intervention.


Dennexin peptides modeled after the homophilic binding sites of the neural cell adhesion molecule (NCAM) promote neuronal survival, modify cell adhesion and impair spatial learning.
Neural cell adhesion molecule (NCAM)-mediated cell adhesion results in activation of intracellular signaling cascades that lead to cellular responses such as neurite outgrowth, neuronal survival, and modulation of synaptic activity associated with cognitive processes. The crystal structure of the immunoglobulin (Ig) 1-2-3 fragment of the NCAM ectodomain has revealed novel mechanisms for NCAM homophilic adhesion. The present study addressed the biological significance of the so called dense zipper formation of NCAM. Two peptides, termed dennexinA and dennexinB, were modeled after the contact interfaces between Ig1 and Ig3 and between Ig2 and Ig2, respectively, observed in the crystal structure. Although the two dennexin peptides differed in amino acid sequence, they both modulated cell adhesion, reflected by inhibition of NCAM-mediated neurite outgrowth. Both dennexins also promoted neuronal survival, and the effect of dennexinA was independent of polysialic acid expression. Consistent with the effect of dennexinA on NCAM-mediated adhesion in vitro, the peptide impaired long-term memory retention in rats in the Morris water maze test. Thus, dennexins are novel site-specific pharmacological tools for elucidation of the role of NCAM in a variety of biological processes under normal and pathological conditions.


The influence of S17 stromal cells and interleukin 7 on B cell development.
A clonal assay was used to study different stimuli involved in the progression of fetal liver B cell precursors to mature B lymphocytes. In this report we replaced fetal liver heterogenous feeder cells by a recombinant growth factor, interleukin 7 (IL 7), and a clonal stromal cell line, S17. Under those conditions we could clone 1 in 10 B220+ B cell precursors from fetal liver and the cells could differentiate to a mitogen-responsive, immunoglobulin-secreting stage. We found that IL 7 stimulates proliferation of B220+ precursors but is not sufficient to support maturation of those precursors to a stage of mitogen responsiveness. We show further that the cell line S17 does not produce IL 7 at functionally detectable level but provides support for B cell maturation. We conclude that this cell line supplies an exogenous stimulus required by B cell precursors to become mature lymphocytes. We describe therefore two stages in pre-B cell development: (a) IL 7-dependent proliferation and (b) S17-dependent maturation to mitogen reactivity. Further studies demonstrate that S17 has a profound effect on B cells by increasing the clonal efficiency of lipopolysaccharide-responsive cells to nearly 1:1 B cell in the spleen of adult C57BL/6 mice.


Chest physiotherapy techniques in neurological intensive care units of India: A survey.
Neurological intensive care units (ICUs) are a rapidly developing sub-specialty of neurosciences. Chest physiotherapy techniques are of great value in neurological ICUs in preventing, halting, or reversing the impairments caused due to neurological disorder and ICU stay. However, chest physiotherapy techniques should be modified to a greater extent in the neurological ICU as compared with general ICUs. The aim of this study is to obtain data on current chest physiotherapy practices in neurological ICUs of India. A tertiary care hospital in Karnataka, India, and cross-sectional survey. A questionnaire was formulated and content validated to assess the current chest physiotherapy practices in neurological ICUs of India. The questionnaire was constructed online and a link was distributed via E-mail to 185 physiotherapists working in neurological ICUs across India. Descriptive statistics. The response rate was 44.3% (n = 82); 31% of the physiotherapists were specialized in cardiorespiratory physiotherapy and 30% were specialized in neurological physiotherapy. Clapping, vibration, postural drainage, aerosol therapy, humidification, and suctioning were used commonly used airway clearance (AC) techniques by the majority of physiotherapists. However, devices for AC techniques such as Flutter, Acapella, and standard positive expiratory pressure devices were used less frequently for AC. Techniques such as autogenic drainage and active cycle of breathing technique are also frequently used when appropriate for the patients. Lung expansion therapy techniques such as breathing exercises, incentive spirometry exercises, and positioning, proprioceptive neuromuscular facilitation of breathing are used by majority of physiotherapists. Physiotherapists in this study were using conventional chest physiotherapy techniques more frequently in comparison to the devices available for AC.


Occurrence of C-terminal residue exclusion in peptide fragmentation by ESI and MALDI tandem mass spectrometry.
By screening a data set of 392 synthetic peptides MS/MS spectra, we found that a known C-terminal rearrangement was unexpectedly frequently occurring from monoprotonated molecular ions in both ESI and MALDI tandem mass spectrometry upon low and high energy collision activated dissociations with QqTOF and TOF/TOF mass analyzer configuration, respectively. Any residue localized at the C-terminal carboxylic acid end, even a basic one, was lost, provided that a basic amino acid such arginine and to a lesser extent histidine and lysine was present in the sequence leading to a fragment ion, usually depicted as (b(n-1) + H(2)O) ion, corresponding to a shortened non-scrambled peptide chain. Far from being an epiphenomenon, such a residue exclusion from the peptide chain C-terminal extremity gave a fragment ion that was the base peak of the MS/MS spectrum in certain cases. Within the frame of the mobile proton model, the ionizing proton being sequestered onto the basic amino acid side chain, it is known that the charge directed fragmentation mechanism involved the C-terminal carboxylic acid function forming an anhydride intermediate structure. The same mechanism was also demonstrated from cationized peptides. To confirm such assessment, we have prepared some of the peptides that displayed such C-terminal residue exclusion as a C-terminal backbone amide. As expected in this peptide amide series, the production of truncated chains was completely suppressed. Besides, multiply charged molecular ions of all peptides recorded in ESI mass spectrometry did not undergo such fragmentation validating that any mobile ionizing proton will prevent such a competitive C-terminal backbone rearrangement. Among all well-known nondirect sequence fragment ions issued from non specific loss of neutral molecules (mainly H(2)O and NH(3)) and multiple backbone amide ruptures (b-type internal ions), the described C-terminal residue exclusion is highly identifiable giving raise to a single fragment ion in the high mass range of the MS/MS spectra. The mass difference between this signal and the protonated molecular ion corresponds to the mass of the C-terminal residue. It allowed a straightforward identification of the amino acid positioned at this extremity. It must be emphasized that a neutral residue loss can be misattributed to the formation of a y(m-1) ion, i.e., to the loss of the N-terminal residue following the a(1)-y(m-1) fragmentation channel. Extreme caution must be adopted when reading the direct sequence ion on the positive ion MS/MS spectra of singly charged peptides not to mix up the attribution of the N- and C-terminal amino acids. Although such peculiar fragmentation behavior is of obvious interest for de novo peptide sequencing, it can also be exploited in proteomics, especially for studies involving digestion protocols carried out with proteolytic enzymes other than trypsin (Lys-N, Glu-C, and Asp-N) that produce arginine-containing peptides.


A novel histone deacetylase inhibitor identified by high-throughput transcriptional screening of a compound library.
Libraries of compounds are increasingly becoming commercially available for the use of individual academic laboratories. A high-throughput system based on a stably integrated transcriptional reporter was used to screen a library of random compounds to identify agents that conferred robust augmentation of a signal transduction pathway. A novel histone deacetylase (HDAC) inhibitor, termed scriptaid, conferred the greatest effect, a 12- to 18-fold augmentation. This facilitation of transcriptional events was generally applicable to exogenous gene constructs, including viral and cellular promoters, different cell lines and reporter genes, and stably integrated and transiently introduced sequences. Scriptaid did not interfere with a further induction provided by stimulation of the cognate signal transduction pathway (transforming growth factor beta/Smad4), which implied the functional independence of ligand-stimulated transcriptional activation and histone acetylation states in this system. Additional insights into this and other signal transduction systems are likely to be afforded through the application of compound screening technologies.


An evaluation of the potential use of Cryptosporidium species as agents for deliberate release.
We evaluated the potential of Cryptosporidium spp. for intentional transmission as a terrorist tactic in asymmetric conflicts in terms of the recognised optimum conditions for biological warfare. Published and widely accepted criteria regarding the optimum conditions for the success of biological warfare based on experience from passive biological warfare research were applied to hypothetical intentional Cryptosporidium spp. transmission. The feasibility of the use of Cryptosporidium spp. transmission for terrorist purposes was established. Particularly on tropical deployments with poor hygiene conditions, such attacks might have a good chance of remaining undetected as a deliberate terrorist attack. Intentional transmission should be suspected in cases of sudden outbreaks of cryptosporidiosis, particularly where adequate food and drinking water hygiene precautions are being enforced. Appropriate diagnostic procedures should be available so that the diagnosis is not missed.


Acute Pancreatitis: Updates for Emergency Clinicians.
Acute pancreatitis is a frequent reason for patient presentation to the emergency department (ED) and the most common gastrointestinal disease resulting in admission. Emergency clinicians are often responsible for the diagnosis and initial management of acute pancreatitis. This review article provides emergency clinicians with a focused overview of the diagnosis and management of pancreatitis. Pancreatitis is an inflammatory process within the pancreas. While the disease is often mild, severe forms can have a mortality rate of up to 30%. The diagnosis of pancreatitis requires two of the following three criteria: epigastric abdominal pain, an elevated lipase, and imaging findings of pancreatic inflammation. The most common etiologies include gallbladder disease and alcohol use. After the diagnosis has been made, it is important to identify underlying etiologies requiring specific intervention, as well as obtain a right upper quadrant ultrasound. The initial management of choice is fluid resuscitation and pain control. Recent data have suggested that more cautious fluid resuscitation in the first 24 h might be more appropriate for some patients. Intravenous opiates are generally safe if used judiciously. Appropriate disposition is a multifactorial decision, which can be facilitated by using Ranson criteria or the Bedside Index of Severity in Acute Pancreatitis score. Complications, though rare, can be severe. Pancreatitis is a potentially deadly disease that commonly presents to most emergency departments. It is important for clinicians to be aware of the current evidence regarding the diagnosis, treatment, and disposition of these patients.


The effect of vaccinating infection during pregnancy and dietary protein supply on the peri-parturient immune response of sheep to infection with Teladorsagia circumcincta and Trichostrongylus colubriformis larvae.
It is well established that dietary protein supply can influence the peri-parturient breakdown of immunity to nematode parasites but there is no information on the importance of exposure to nematode larvae during pregnancy for this response. We investigated this by exposing housed pregnant sheep, scanned as carrying two lambs, to a vaccinating infection with a trickle mixed infection of Teladorsagia circumcincta and Trichostrongylus colubriformis larvae (L3) or to no infection during weeks - 9 to - 4 relative to parturition. At the beginning of week - 3 all sheep were treated with anthelmintic to remove any vaccinating worm burden and from week - 2 to week +6 received a trickle challenge infection with the same nematodes. Within each vaccinating treatment there were two nutritional treatments (no. = 20 per subgroup) designed to provide 1.5 or 1.0 and 1.3 or 0.8 of metabolisable protein (MP) requirement during pregnancy and lactation, respectively. Five ewes were necropsied during weeks +1 and +3 to measure worm burdens and mucosal inflammatory cells and the remainder maintained until week +6. Serum levels of total, IgA and IgE antibodies against L3 antigen of each nematode were measured.Scanning errors and lamb losses resulted in some ewes carrying and/or rearing only one lamb. Numbers of lambs reared was therefore introduced as a treatment effect. Vaccinating infection delayed the peri-parturient rise in faecal egg count (FEC) by an average of 2 weeks but its effect on FEC during the first 6 weeks of lactation was smaller and less persistent than that of dietary MP supply and single- v. twin-suckling.Populations of both nematodes were lower in association with high MP supply, vaccination and single suckling. These changes were associated with increases in numbers of mucosal mast cells (MMC) as a result of both increased MP supply and vaccination. Evidence for a more rapid return of host ability to limit populations of the abdominal nematode T. circumcincta than of the intestinal nematode T. colubriformis was associated with fewer eosinophils and more globule leucocytes (GL) in abomasal than in intestinal tissue.None of the serum antibody isotypes was affected by dietary protein supply. Total and IgA antibodies were maintained by a current larval (vaccinating) intake. IgA titres, however, increased progressively during pregnancy, especially in twin-bearing ewes. IgE titres appeared to be sensitive primarily to the reproductive cycle itself, peaking around parturition.This work supports the conclusion that availability of MP supply influences the recruitment and activity of cells of the immune armoury of the gastro-intestinal tract to nematode parasites. The precise outcome may differ with site and/or nematode species.


Brain substrates of social decision-making in dual diagnosis: cocaine dependence and personality disorders.
Cocaine dependence frequently co-occurs with personality disorders, leading to increased interpersonal problems and greater burden of disease. Personality disorders are characterised by patterns of thinking and feeling that divert from social expectations. However, the comorbidity between cocaine dependence and personality disorders has not been substantiated by measures of brain activation during social decision-making. We applied functional magnetic resonance imaging to compare brain activations evoked by a social decision-making task-the Ultimatum Game-in 24 cocaine dependents with personality disorders (CDPD), 19 cocaine dependents without comorbidities and 19 healthy controls. In the Ultimatum Game participants had to accept or reject bids made by another player to split monetary stakes. Offers varied in fairness (in fair offers the proposer shares ~50 percent of the money; in unfair offers the proposer shares <30 percent of the money), and participants were told that if they accept both players get the money, and if they reject both players lose it. We contrasted brain activations during unfair versus fair offers and accept versus reject choices. During evaluation of unfair offers CDPD displayed lower activation in the insula and the anterior cingulate cortex and higher activation in the lateral orbitofrontal cortex and superior frontal and temporal gyri. Frontal activations negatively correlated with emotion recognition. During rejection of offers CDPD displayed lower activation in the anterior cingulate cortex, striatum and midbrain. Dual diagnosis is linked to hypo-activation of the insula and anterior cingulate cortex and hyper-activation of frontal-temporal regions during social decision-making, which associates with poorer emotion recognition.


A role for PS integrins in morphological growth and synaptic function at the postembryonic neuromuscular junction of Drosophila.
A family of three position-specific (PS) integrins are expressed at the Drosophila neuromuscular junction (NMJ): a beta subunit ((betaPS), expressed in both presynaptic and postsynaptic membranes, and two alpha subunits (alphaPS1, alphaPS2), expressed at least in the postsynaptic membrane. PS integrins appear at postembryonic NMJs coincident with the onset of rapid morphological growth and terminal type-specific differentiation, and are restricted to type I synaptic boutons, which mediate fast, excitatory glutamatergic transmission. We show that two distinctive hypomorphic mutant alleles of the beta subunit gene myospheroid (mys(b9) and mys(ts1)), differentially affect betaPS protein expression at the synapse to produce distinctive alterations in NMJ branching, bouton formation, synaptic architecture and the specificity of synapse formation on target cells. The mys(b9) mutation alters betaPS localization to cause a striking reduction in NMJ branching, bouton size/number and the formation of aberrant 'mini-boutons', which may represent a developmentally arrested state. The mys(ts1) mutation strongly reduces betaPS expression to cause the opposite phenotype of excessive synaptic sprouting and morphological growth. NMJ function in these mutant conditions is altered in line with the severity of the morphological aberrations. Consistent with these mutant phenotypes, transgenic overexpression of the betaPS protein with a heat-shock construct or tissue-specific GAL4 drivers causes a reduction in synaptic branching and bouton number. We conclude that betaPS integrin at the postembryonic NMJ is a critical determinant of morphological growth and synaptic specificity. These data provide the first genetic evidence for a functional role of integrins at the postembryonic synapse.


Managing Noncommunicable Diseases in an African Community: Effects, Compliance, and Barriers to Participation in a 4-Week Exercise Intervention.
To determine the compliance, barriers, and effects of participation in a 4-week exercise intervention aimed at reducing risk factors for noncommunicable diseases among community-dwelling adults from a low-resourced area of South Africa. An exercise program and associated pre-posttest were performed by 76 participants (men, n = 26 and women, n = 50) aged 35 to 65 years. Baseline and end tests included height, weight, hip and waist circumference, heart rate, blood pressure, glucose, cholesterol, quality of life, and cardiorespiratory fitness measurements. The intervention consisted of 3 days/week combined aerobic and resistance exercise at an intensity of 70% heart rate reserved as determined at baseline. Compliance and barriers to participation were determined post-intervention by means of attendance registers and interviews. ANCOVA with adjustment for pretest was performed for all repeated variables. The Cronbach's alpha coefficients for exercise benefits were 0.81 and for barriers 0.84. Of the 26 men (40.8 ± 5.45 years) and 50 women (43.6 ± 7.8 years) recruited, 54 completed the intervention (71% compliance). The 4-week aerobic exercise intervention significantly reduced body mass, rate of perceived exertion, and mental components summary in men, and body mass, body mass index, VO2max, rate of perceived exertion, glucose, physical components summary, and mental components summary in women. Participants reported that the exercise milieu as a major barrier to exercise compliance while the interviews reported lack of time. A 1-month exercise intervention elucidated positive changes in risk factors for noncommunicable diseases in a low-resource community. A drop-out rate of 29% in this study is consistent with other exercise intervention trials. Exploration of the reported barriers may be useful for planning to increase compliance with future programs.


Developmental evolutionary biology of the vertebrate ear: conserving mechanoelectric transduction and developmental pathways in diverging morphologies.
This brief overview shows that a start has been made to molecularly dissect vertebrate ear development and its evolutionary conservation to the development of the insect hearing organ. However, neither the patterning process of the ear nor the patterning process of insect sensory organs is sufficiently known at the moment to provide more than a first glimpse. Moreover, hardly anything is known about otocyst development of the cephalopod molluscs, another triploblast lineage that evolved complex 'ears'. We hope that the apparent conserved functional and cellular components present in the ciliated sensory neurons/hair cells will also be found in the genes required for vertebrate ear and insect sensory organ morphogenesis (Fig. 3). Likewise, we expect that homologous pre-patterning genes will soon be identified for the non-sensory cell development, which is more than a blocking of neuronal development through the Delta/Notch signaling system. Generation of the apparently unique ear could thus represent a multiplication of non-sensory cells by asymmetric and symmetric divisions as well as modification of existing patterning process by implementing novel developmental modules. In the final analysis, the vertebrate ear may come about by increasing the level of gene interactions in an already existing and highly conserved interactive cascade of bHLH genes. Since this was apparently achieved in all three lineages of triploblasts independently (Fig. 3), we now need to understand how much of the morphogenetic cascades are equally conserved across phyla to generate complex ears. The existing mutations in humans and mice may be able to point the direction of future research to understand the development of specific cell types and morphologies in the formation of complex arthropod, cephalopod, and vertebrate 'ears'.


Spectral characterization of integrated acousto-optic tunable filters by means of laser frequency modulation spectroscopy.
The spectral characteristics of an integrated acousto-optic tunable filter (AOTF) as well as its responsivity to the rf driving signal and sensitivity to temperature changes are experimentally investigated and quantified using a diode-laser-based interrogation system. A spectroscopic technique, exploiting the rf frequency modulation of the laser beam and the phase-sensitive detection of the AOTF transmission, has been used for this purpose. That allows for the generation of a dispersivelike signal, which serves as a reference for tracking any wavelength change of the filter's peak with high resolution. The possibility of using the integrated AOTF as a spectrum analyzer with this interrogation scheme for fiber Bragg grating (FBG) strain sensing is also discussed.


Early Adolescent Trajectories of Impulsiveness and Sensation Seeking in Children of Fathers with Histories of Alcohol and Other Substance Use Disorders.
Problem substance use often begins in adolescence. This vulnerability likely stems, at least partially, from relatively rapid increases in sensation seeking occurring in early to mid-adolescence and more gradual improvements in impulse control occurring through later adolescence. Better understanding how these processes develop in high-risk youth may lead to enhanced substance use disorder treatment and prevention strategies. We characterized trajectories of self-reported impulsivity and sensation seeking in 305 FH+ youths who at minimum had a father with a history of alcohol or other drug use disorders and 81 youths with no family histories of substance use disorders (FH-). Assessments started at ages 10 to 12 and continued at 6-month intervals for up to 42 months. In addition, a subset of 58 FH+ youths who began alcohol or other drug use before age 15 (FH+ Users) were compared to 58 FH+ propensity-matched adolescents who did not initiate substance use before age 15 (FH+ Non-Users). Compared to FH- youths at preadolescence, FH+ youths reported higher general impulsivity and higher impulsivity related to poor planning and attention. Over time, there were no differential effects of FH status on changes in impulsivity or sensation seeking across adolescence. FH+ Users had smaller decreases in general impulsivity and impulsivity related to restlessness and fidgeting across adolescence than FH+ Non-Users. FH+ Users also had greater increases in sensation seeking across adolescence than FH+ Non-Users. Increased impulsivity in FH+ youths may make them less able to regulate sensation seeking drives that peak in adolescence, which may contribute to their high risk for developing substance use disorders. Additionally, FH+ adolescents who initiate early use may be at increased risk in part due to increased impulsivity coupled with greater increases in sensation seeking.


Inactivating mutations in NPC1L1 and protection from coronary heart disease.
Ezetimibe lowers plasma levels of low-density lipoprotein (LDL) cholesterol by inhibiting the activity of the Niemann-Pick C1-like 1 (NPC1L1) protein. However, whether such inhibition reduces the risk of coronary heart disease is not known. Human mutations that inactivate a gene encoding a drug target can mimic the action of an inhibitory drug and thus can be used to infer potential effects of that drug. We sequenced the exons of NPC1L1 in 7364 patients with coronary heart disease and in 14,728 controls without such disease who were of European, African, or South Asian ancestry. We identified carriers of inactivating mutations (nonsense, splice-site, or frameshift mutations). In addition, we genotyped a specific inactivating mutation (p.Arg406X) in 22,590 patients with coronary heart disease and in 68,412 controls. We tested the association between the presence of an inactivating mutation and both plasma lipid levels and the risk of coronary heart disease. With sequencing, we identified 15 distinct NPC1L1 inactivating mutations; approximately 1 in every 650 persons was a heterozygous carrier for 1 of these mutations. Heterozygous carriers of NPC1L1 inactivating mutations had a mean LDL cholesterol level that was 12 mg per deciliter (0.31 mmol per liter) lower than that in noncarriers (P=0.04). Carrier status was associated with a relative reduction of 53% in the risk of coronary heart disease (odds ratio for carriers, 0.47; 95% confidence interval, 0.25 to 0.87; P=0.008). In total, only 11 of 29,954 patients with coronary heart disease had an inactivating mutation (carrier frequency, 0.04%) in contrast to 71 of 83,140 controls (carrier frequency, 0.09%). Naturally occurring mutations that disrupt NPC1L1 function were found to be associated with reduced plasma LDL cholesterol levels and a reduced risk of coronary heart disease. (Funded by the National Institutes of Health and others.).


Single-cell analysis reveals a stem-cell program in human metastatic breast cancer cells.
Despite major advances in understanding the molecular and genetic basis of cancer, metastasis remains the cause of >90% of cancer-related mortality. Understanding metastasis initiation and progression is critical to developing new therapeutic strategies to treat and prevent metastatic disease. Prevailing theories hypothesize that metastases are seeded by rare tumour cells with unique properties, which may function like stem cells in their ability to initiate and propagate metastatic tumours. However, the identity of metastasis-initiating cells in human breast cancer remains elusive, and whether metastases are hierarchically organized is unknown. Here we show at the single-cell level that early stage metastatic cells possess a distinct stem-like gene expression signature. To identify and isolate metastatic cells from patient-derived xenograft models of human breast cancer, we developed a highly sensitive fluorescence-activated cell sorting (FACS)-based assay, which allowed us to enumerate metastatic cells in mouse peripheral tissues. We compared gene signatures in metastatic cells from tissues with low versus high metastatic burden. Metastatic cells from low-burden tissues were distinct owing to their increased expression of stem cell, epithelial-to-mesenchymal transition, pro-survival, and dormancy-associated genes. By contrast, metastatic cells from high-burden tissues were similar to primary tumour cells, which were more heterogeneous and expressed higher levels of luminal differentiation genes. Transplantation of stem-like metastatic cells from low-burden tissues showed that they have considerable tumour-initiating capacity, and can differentiate to produce luminal-like cancer cells. Progression to high metastatic burden was associated with increased proliferation and MYC expression, which could be attenuated by treatment with cyclin-dependent kinase (CDK) inhibitors. These findings support a hierarchical model for metastasis, in which metastases are initiated by stem-like cells that proliferate and differentiate to produce advanced metastatic disease.


liqDB: a small-RNAseq knowledge discovery database for liquid biopsy studies.
MiRNAs are important regulators of gene expression and are frequently deregulated under pathologic conditions. They are highly stable in bodily fluids which makes them feasible candidates to become minimally invasive biomarkers. In fact, several studies already proposed circulating miRNA-based biomarkers for different types of neoplastic, cardiovascular and degenerative diseases. However, many of these studies rely on small RNA sequencing experiments that are based on different RNA extraction and processing protocols, rendering results incomparable. We generated liqDB, a database for liquid biopsy small RNA sequencing profiles that provides users with meaningful information to guide their small RNA liquid biopsy research and to overcome technical and conceptual problems. By means of a user-friendly web interface, miRNA expression profiles from 1607 manually annotated samples can be queried and explored at different levels. Result pages include downloadable expression matrices, differential expression analysis, most stably expressed miRNAs, cluster analysis and relevant visualizations by means of boxplots and heatmaps. We anticipate that liqDB will be a useful tool in liquid biopsy research as it provides a consistently annotated large compilation of experiments together with tools for reproducible analysis, comparison and hypothesis generation. LiqDB is available at http://bioinfo5.ugr.es/liqdb.


Complete Genome Sequence of the Bioluminescent Marine Bacterium Vibrio harveyi ATCC 33843 (392 [MAV]).
Vibrio harveyi is a Gram-negative marine γ-proteobacterium that is known to be a formidable pathogen of aquatic animals and is a model organism for the study of bacterial bioluminescence and quorum sensing. In this report, we describe the complete genome sequence of the most studied strain of this species: V. harveyi ATCC 33843 (392 [MAV]).


[Prevalence and costs of chronicity and multimorbidity in the population covered by the Basque public telecare service].
The public telecare service in the Basque Autonomous Community (betiON) is a service offered by the Department of Employment and Social Policy of the Basque Government and offers telephone support to elderly and disabled populations, providing communication with an assistance centre. This connection with health care devices makes it possible to implement shared information systems with health and social data, which enable a characterization of the population attended to and the development of integrated models for health and social provision. To describe the distribution of chronic diseases and multimorbidity among users of betiON, to estimate the cost of their health care and to compare these indicators with those of the general population. We extracted information from betiON and public healthcare databases. We characterized the telecare population above 65 years (N = 23.742) and compared it with the total population in the Basque Autonomous Community (N = 2.262.707) with respect to variables in demography socio-economic burden, costs and ambulatory care conditions. In the telecare population, the average age is 83 years, 80% are women, 55.1% live alone and reside in poorer neighbourhoods. Forty-two point five per cent of males and 36.5% of women are affected by five or more chronic conditions, averaging 4.06 pathologies per person. They also show higher healthcare costs than the general population. Telecare users have a higher disease burden, greater need for healthcare and unfavourable social conditions. Telecare provides an opportunity for innovation and intervention, through the coordination of social and healthcare services.


Levels of detection of tumor cells in human bone marrow with or without prior culture.
With an in vitro culture technique combined with light microscopy, immunocytochemistry and molecular probing, we previously detected occult tumor cells in histologically-normal human bone marrow harvested for autologous transplantation. In this study, we mixed known numbers of malignant lymphoid (Raji and CEM) or breast cancer (MCF-7) cells with normal human bone marrow cells to determine the levels at which tumor cells can be detected before and after culture. Cytocentrifuge preparations were made before culture and after 2 or more weeks of culture and examined by light microscopy. We detected contaminating lymphoma cells at a level of more than 5% before culture, and at a level of 0.01% after culture for 2 or more weeks in 2% human lymphocyte conditioned medium. Before culture, we detected MCF-7 cells at a level of 0.001% using glucose oxidase immunocytochemical staining techniques; these cells were detected at a level of 0.00001% after culture. Since, of necessity, these calibrations rations were performed using cell lines, it is likely that these results overestimate the absolute sensitivity of these methods for detection of tumor cells in patient samples. We found the glucose oxidase immunocytochemical method more specific for detecting occult tumor cells in bone marrow than the immunoperoxidase staining method because of the absence of non-specific staining arising from endogenous peroxidase in bone marrow cells which makes the interpretation of the latter difficult. We conclude that culture techniques can increase the sensitivity of detection of occult tumor cells in human bone marrow about 100-fold.


Neural defects and cardiac arrhythmia in fish larvae following embryonic exposure to 2,2',4,4'-tetrabromodiphenyl ether (PBDE 47).
Polybrominated diphenyl ethers (PBDEs) are added to plastics, polyurethane foam, and textiles as a flame retardant. While PBDEs play a key role in reducing loss of human life and property from fires, these flame retardants have become pervasive organic contaminants in the environment and in the tissues of fish, birds, marine mammals, and humans. Levels of PBDEs in wildlife and humans continue to rise, raising concerns about potential ecological and health risks associated with exposure to these chemicals. Nevertheless, there is little currently known about the toxicological effects of PBDE exposure. Here, we examined the developmental toxicity of the PBDE congener 2,2',4,4'-tetrabromodiphenyl ether (PBDE 47) using the zebrafish (Danio rerio) as an ontogenetic model. Zebrafish embryos were exposed continuously to dissolved phase PBDE 47 (100-5000 microg/l) beginning 3-5 h post-fertilization (hpf). Fish treated with the highest concentrations of PBDE 47 delayed hatching, had reduced growth post-hatching, and displayed an abnormal dorsal curvature of the body with flexion at the hindbrain. By 96h post-fertilization larvae exposed to PBDE 47 had significant tachycardia, which progressed into atrioventricular block arrhythmias. Microinjection of fluorescent dye into the hindbrain ventricle revealed that cerebrospinal fluid in the neural tube and brain ventricles flowed more slowly in fish larvae exposed to PBDE 47, a likely etiology for the dorsal curvature. Similar, though much less pronounced, developmental toxicity also occurred in larvae exposed to PBDE 47 only for a 20h period during early embryogenesis (3-23 hpf), suggesting that PBDEs incorporated in lipid of the egg are bioavailable and cause toxicity later in life. Taken together, this work indicates that exposure to PBDE 47 can cause morphological abnormalities, impair cardiovascular function and cerebrospinal fluid flow, and provides a tractable starting point for using the zebrafish model to explore molecular mechanisms of PBDE toxicity.


Liquid-phase membrane extraction of targeted pesticides from manufacturing wastewaters in a hollow fibre contactor with feed-stream recycle.
A two-phase membrane extraction in a hollow fibre contactor with feed-stream recycle was applied to remove selected pesticides (tebufenozide, linuron, imidacloprid, acetamiprid and dimethoate) from their mixed aqueous solutions. The contactor consisted of 50 polypropylene hollow fibres impregnated with 5% tri-n-octylphosphine oxide in di-n-hexyl ether. For low-polar pesticides with log P ≥ 2 (tebufenozide and linuron), the maximum removal efficiency increased linearly from 85% to 96% with increasing the feed flow rate. The maximum removal efficiencies of more polar pesticides were significantly higher under feed recirculation (86%) than in a continuous single-pass operation (30%). It was found from the Wilson's plot that the mass transfer resistance of the liquid membrane can be neglected for low-polar pesticides. The pesticide removals from commercial formulations were similar to those from pure pesticide solutions, indicating that built-in adjuvants did not affect the extraction process.