Gametogenesis of intergroup hybrids of hemiclonal frogs.
European water frog hybrids Rana esculenta (R. ridibundaxR. lessonae) reproduce hemiclonally, by hybridogenesis: in the germ line they exclude the genome of one parental species and produce haploid gametes with an unrecombined genome of the other parental species. In the widespread L-E population system, both sexes of hybrids (E) coexist with R. lessonae (L). They exclude the lessonae genome and produce ridibunda gametes. In the R-E system, hybrid males coexist with R. ridibunda (R); they exclude either their ridibunda or their lessonae genome and produce sperm with a lessonae or with a ridibunda genome or a mixture of both kinds of sperm. We examined 13 male offspring, 12 of which were from crosses between L-E system and R-E system frogs. All were somatically hybrid. With one exception, they excluded the lessonae genome in the germ line and subsequently endoreduplicated the ridibunda genome. Spermatogonial metaphases contained a haploid or a diploid number of ridibunda chromosomes, identified through in situ hybridization to a satellite DNA marker, and by spermatocyte I metaphases containing a haploid number of ridibunda bivalents. The exception, an F1 hybrid between L-E system R. lessonae and R-E system R. ridibunda, was not hybridogenetic, showed no genome exclusion, and evidenced a disturbed gametogenesis resulting from the combination of two heterospecific genomes. None of the hybridogenetic hybrids showed any cell lines excluding the ridibunda genome, the pattern most frequent in hybrids of the R-E system, unique to that system, and essential for its persistence. A particular combination of R-E system lessonae and R-E system ridibunda genomes seems necessary to induce the R-E system type of hemiclonal gametogenesis.


Single cell analysis of spondyloarthritis regulatory T cells identifies distinct synovial gene expression patterns and clonal fates.
Regulatory T cells (Tregs) play an important role in controlling inflammation and limiting autoimmunity, but their phenotypes at inflammatory sites in human disease are poorly understood. We here analyze the single-cell transcriptome of >16,000 Tregs obtained from peripheral blood and synovial fluid of two patients with HLA-B27+ ankylosing spondylitis and three patients with psoriatic arthritis, closely related forms of inflammatory spondyloarthritis. We identify multiple Treg clusters with distinct transcriptomic profiles, including, among others, a regulatory CD8+ subset expressing cytotoxic markers/genes, and a Th17-like RORC+ Treg subset characterized by IL-10 and LAG-3 expression. Synovial Tregs show upregulation of interferon signature and TNF receptor superfamily genes, and marked clonal expansion, consistent with tissue adaptation and antigen contact respectively. Individual synovial Treg clones map to different clusters indicating cell fate divergence. Finally, we demonstrate that LAG-3 directly inhibits IL-12/23 and TNF secretion by patient-derived monocytes, a mechanism with translational potential in SpA. Our detailed characterization of Tregs at an important inflammatory site illustrates the marked specialization of Treg subpopulations.


Use of combination neonatal prophylaxis for the prevention of mother-to-child transmission of HIV infection in European high-risk infants.
To evaluate use of combination neonatal prophylaxis (CNP) in infants at high risk for mother-to-child transmission (MTCT) of HIV in Europe and investigate whether CNP is more effective in preventing MTCT than single drug neonatal prophylaxis (SNP). Individual patient-data meta-analysis across eight observational studies. Factors associated with CNP receipt and with MTCT were explored by logistic regression using data from nonbreastfed infants, born between 1996 and 2010 and at high risk for MTCT. In 5285 mother-infant pairs, 1463 (27.7%) had no antenatal or intrapartum antiretroviral prophylaxis, 915 (17.3%) had only intrapartum prophylaxis and 2907 (55.0%) mothers had detectable delivery viral load despite receiving antenatal antiretroviral therapy. Any neonatal prophylaxis was administered to 4623 (87.5%) infants altogether; 1105 (23.9%) received CNP. Factors significantly associated with the receipt of CNP were later calendar birth year, no elective caesarean section, maternal CD4 cell count less than 200 cells/μl, maternal delivery viral load more than 1000 copies/ml, no antenatal antiretroviral therapy, receipt of intrapartum single-dose nevirapine and cohort. After adjustment, absence of neonatal prophylaxis was associated with higher risk of MTCT compared to neonatal prophylaxis [adjusted odds ratio (aOR) 2.29; 95% confidence interval (95% CI) 1.46-2.59; P < 0.0001]. Further, there was no association between CNP and MTCT compared to SNP (aOR 1.41; 95% CI 0.97-2.5; P = 0.07). In this European population, CNP use is increasing and associated with presence of MTCT risk factors. The finding of no observed difference in MTCT risk between one drug and CNP may reflect residual confounding or the fact that CNP may be effective only in a subgroup of infants rather than the whole population of high-risk infants.


Drawn to Distraction: Anxiety Impairs Neural Suppression of Known Distractor Features in Visual Search.
When searching for a target, it is possible to suppress the features of a known distractor. This suppression may prevent distractor processing altogether or only after the distractor initially captures attention (i.e., search and destroy). However, suppression may be impaired in individuals with attentional control deficits, such as in high anxiety. In this study (n = 48), we used ERPs to examine the time course of attentional enhancement and suppression when participants were given pretrial information about target or distractor features. Consistent with our hypothesis, we found that individuals with higher levels of anxiety had lower neural measures of suppressing the template-matching distractor, instead showing enhanced processing. These findings indicate that individuals with anxiety are more likely to use a search-and-destroy mechanism of negative templates-highlighting the importance of attentional control abilities in distractor-guided search.


Exposure to crosslinked IgD induces receptors for IgD on T cells in vivo and in vitro.
IgD is a surface immunoglobulin, which is coexpressed with IgM on greater than 90% of mature B cells, but its levels in serum are extremely low compared to those of IgM. It role as a surface receptor has been reemphasized by our recent findings that IgD receptors are induced on helper T cells by exposure to IgD and that such cells have immunoaugmenting properties. The present study shows that crosslinking of soluble IgD or of monomeric cell surface IgD is required and sufficient for the induction of T cells bearing receptors for IgD, both in vivo and in vitro. Effective IgD crosslinking in this respect can be obtained with antigen or with heterologous and immunogenic as well as nonimmunogenic allotype-specific anti-IgD. These results reinforce the concept that the induction of T cells bearing receptors for IgD is an integral component of the normal immune response.


Magnetic resonance imaging features of sinonasal disorders in horses.
Diseases of paranasal sinuses and nasal passages in horses can be a diagnostic challenge because of the complex anatomy of the head and limitations of many diagnostic modalities. Our hypothesis was that magnetic resonance (MR) imaging would provide excellent anatomical detail and soft tissue resolution, and would be accurate in the diagnosis of diseases of the paranasal sinuses and nasal passages in horses. Fourteen horses were imaged. Inclusion criteria were lesions located to the sinuses or nasal passages that underwent MR imaging and subsequent surgical intervention and/or histopathologic examination. A low field, 0.3 tesla open magnet was used. Sequences in the standard protocol were fast spin echo T2 sagittal and transverse, spin echo T1 transverse, short-tau inversion recovery (STIR) dorsal, gradient echo 3D T1 MPR dorsal (plain and contrast enhanced), spin echo T1 fatsat (contrast enhanced). Mean scan time to complete the examination was 53 min (range 39-99 min). Lesions identified were primary or secondary sinusitis (six horses), paranasal sinus cyst (four horses), progressive ethmoid hematoma (two horses), and neoplasia (two horses). The most useful sequences were fast spin echo T2 transverse and sagittal, STIR dorsal and FE3D MPR (survey and contrast enhanced). Fluid accumulation, mucosal thickening, presence of encapsulated contents, bone deformation, and thickening were common findings observed in MR imaging. In selected horses, magnetic resonance imaging is a useful tool in diagnosing lesions of the paranasal sinuses and nasal passages.


Precocious puberty and a sellar mass.
Precocious puberty in a child less than 3 years of age is usually gonadotropin dependent and is often associated with extrasellar central nervous system disorders. The present report describes the case of a 2½-year-old girl who presented with gonadotropin-independent precocious puberty and had a sellar mass. She was referred for neurosurgical intervention to our institute. On investigation, she was found to have primary hypothyroidism of autoimmune aetiology, which is rarely associated with precocious puberty and sellar mass due to thyrotroph hyperplasia. She was successfully treated with levothyroxine, and with this secondary sexual characteristics progressively regressed and the sellar mass disappeared.


Light traffic: photo-crosslinking a novel transport system.
The Tat protein transporter is found in the membranes of many bacteria and in plant chloroplasts. This highly unusual transport machine moves folded and often oligomeric substrate proteins across energy-conserving membranes. A recent paper reports the first use of a photo-crosslinking approach to dissect the early recognition events between the transporter and its substrate.


Using daily diagnostic quality images to validate planning margins for prostate interfractional variations
The purpose of this study is to use the same diagnostic-quality verification and planning CTs to validate planning margin account for residual interfractional variations with image-guided soft tissue alignment of the prostate. For nine pros-tate cancer patients treated with IMRT to 78 Gy in 39 fractions, daily verification CT-on-rails images of the first seven and last seven fractions (n = 126) were retrospectively selected for this study. On these images, prostate, bladder, and rectum were delineated by the same attending physician. Clinical plans were cre-ated with a margin of 8 mm except for 5 mm posteriorly, referred to as 8/5mm. Three additional plans were created for each patient with the margins of 6/4 mm, 4/2mm, and 2 mm uniform. These plans were subsequently applied to daily images and radiation doses were recalculated. The isocenters of these plans were placed according to clinical online shifts, which were based on soft tissue alignment to the prostate. Retrospective offline shifts by aligning prostate contours were com-pared to online shifts. The resultant daily target dose was analyzed using D99, the percentage of the prescription dose received by 99% of CTV. The percent of blad-der volume receiving 65 Gy (V65Gy) and rectum V70Gy were also analyzed. After interfractional correction, using CTV D99 > 97% criteria, 8/5 mm, 6/4 mm, 4/2 mm, and 2 mm planning margins met the CTV dose coverage in 95%, 91%, 65%, and 53% of the 126 fractions with online shifts, and 99%, 98%, 85%, and 68% with offline shifts. The rectum V70Gy and bladder V65Gy were significantly decreased at each level of margin reduction (p < 0.05). With daily diagnostic quality imaging-guidance, the interfractional planning margin may be reduced from 8/5mm to 6/4 mm. The residual interfractional uncertainties most likely stem from prostate rotation anddeformation.


Illicit and injecting drug use among Indigenous young people in urban, regional and remote Australia.
To examine patterns of illicit drug use among Australian Indigenous young people, identify correlates of frequent use separately in urban, regional and remote settings and characterise those who inject. Cross-sectional design at 40 Indigenous events. Self-complete surveys were administered to Indigenous people aged 16-29 years using mobile devices. 2,877 participants completed the survey. One in five reported using cannabis at least weekly in the previous year, but the use of other drugs was less prevalent. Patterns of drug use were largely similar across regions, although more participants in urban and regional areas reported using ecstasy (12% vs 11% vs 5%) and cocaine (6% vs 3% vs 1%) and more reported weekly cannabis use (18% vs 22% vs 14%). Injecting was rare (3%) but those who did inject reported a high incidence of needle sharing (37%). Methamphetamine (37%), heroin (36%) and methadone (26%) were the most commonly injected drugs, and injecting was related to prison experience (AOR 5.3 95% CI 2.8-10.0). Attention is needed in relation to cannabis use, particularly among those Indigenous young people living in regional and urban settings. Also, although injecting is uncommon, it is associated with prison involvement. Priority must be given to reducing the numbers of Indigenous youth entering justice settings, delaying the age at first entry to justice settings, and reducing the risk of BBV acquisition while in custody through, for example, prison-based NSP, BBV education, and Indigenous-specific treatment that emphasises connection to country and culture. [Bryant J, Ward J, Wand H, Byron K, Bamblett A, Waples-Crowe P, Betts S, Coburn T, Delaney-Thiele D, Worth H, Kaldor J, Pitts M. Illicit and injecting drug use among Indigenous young people in urban, regional and remote Australia. Drug Alcohol Rev 2016;35:447-455].


Complexity behind CO2 capture on NH2-MIL-53(Al).
Some Metal Organic Frameworks (MOFs) show excellent performance in extracting carbon dioxide from different gas mixtures. The origin of their enhanced separation ability is not clear yet. Herein, we present a combined experimental and theoretical study of the amino-functionalized MIL-53(Al) to elucidate the mechanism behind its unusual high efficiency in CO(2) capture. Spectroscopic and DFT studies point out only an indirect role of amine moieties. In contrast to other amino-functionalized CO(2) sorbents, no chemical bond between CO(2) and the NH(2) groups of the structure is formed. We demonstrate that the functionalization modulates the "breathing" behavior of the material, that is, the flexibility of the framework and its capacity to alter the structure upon the introduction of specific adsorbates. The absence of strong chemical interactions with CO(2) is of high importance for the overall performance of the adsorbent, since full regeneration can be achieved within minutes under very mild conditions, demonstrating the high potential of this type of adsorbents for PSA like systems.


Steric effects of ions in the charge-related wetting phenomena.
Steric effects of ions on the charge-related wetting phenomena are studied. Along with a general treatment, three specific problems in two-dimensional system are considered: a droplet on an electrode, a droplet on a charged surface, and an electrowetting phenomenon on a dielectric. For computation of wetting tension, the electromechanical approach is adopted with the principle of mechanical force balance for each phase. The modified Poisson-Boltzmann equation, which was originally proposed by Bikerman [Philos. Mag. 33, 384 (1942)], is adopted for the analysis of the steric effects. It is found that the steric hindrance reduces significantly both the osmotic pressure and the electrical stress near the triple contact line. This reduction results in a considerable decrease in the wetting tension when the ratio of the capacitance per unit area of the electrical double layer to that of the dielectric layer is small.


Extended-spectrum-beta-lactamase producing bacteria related urinary tract infection in renal transplant recipients and effect on allograft function.
Urinary tract infection (UTI) is a well-recognized early complication in renal transplant recipients (RTR) and can have significant bearing on their outcome. The recent rise in incidence of extended spectrum beta lactamase (ESBL) producing bacteria causing UTI among RTR poses new and significant challenges in terms of management and outcome. Our aim is to analyze the effect of ESBL producing bacteria causing UTI in these patients and its impact on allograft function. We reviewed the medical records of 147 RTR who were followed at a tertiary care hospital affiliated transplant center between January 2007 and May 2013 and noted five RTR who developed episodes of ESBL producing bacteria related UTI during follow up. Multiple patient characteristics including demographics, immunosuppression, recurrences, allograft function and outcome were analyzed. Five patients (3.4%) out of 147 had ESBL producing bacteria related UTI. We found all patients to be above 60 years of age, with three out of five being females, and all five patients had diabetes mellitus. We identified a total of 37 episodes of UTI among these five patients during this period. Two of these patients had elevated creatinine values during the episodes of UTI and three of them developed bacteremia. Of the five patients, four of them had a favorable outcome except for one patient who developed persistent allograft dysfunction. RTR are at a higher risk for developing ESBL producing bacteria associated UTI. Early diagnosis along with appropriate and judicious use of antibiotics will ensure long term success in allograft and patient outcome.


The empirical status of cognitive-behavioral therapy: a review of meta-analyses.
This review summarizes the current meta-analysis literature on treatment outcomes of CBT for a wide range of psychiatric disorders. A search of the literature resulted in a total of 16 methodologically rigorous meta-analyses. Our review focuses on effect sizes that contrast outcomes for CBT with outcomes for various control groups for each disorder, which provides an overview of the effectiveness of cognitive therapy as quantified by meta-analysis. Large effect sizes were found for CBT for unipolar depression, generalized anxiety disorder, panic disorder with or without agoraphobia, social phobia, posttraumatic stress disorder, and childhood depressive and anxiety disorders. Effect sizes for CBT of marital distress, anger, childhood somatic disorders, and chronic pain were in the moderate range. CBT was somewhat superior to antidepressants in the treatment of adult depression. CBT was equally effective as behavior therapy in the treatment of adult depression and obsessive-compulsive disorder. Large uncontrolled effect sizes were found for bulimia nervosa and schizophrenia. The 16 meta-analyses we reviewed support the efficacy of CBT for many disorders. While limitations of the meta-analytic approach need to be considered in interpreting the results of this review, our findings are consistent with other review methodologies that also provide support for the efficacy CBT.


Long-term effects of caffeine therapy for apnea of prematurity on sleep at school age.
Apnea of prematurity is a common condition that is usually treated with caffeine, an adenosine receptor blocker that has powerful influences on the central nervous system. However, little is known about the long-term effects of caffeine on sleep in the developing brain. We hypothesized that neonatal caffeine use resulted in long-term abnormalities in sleep architecture and breathing during sleep. A total of 201 ex-preterm children aged 5-12 years who participated as neonates in a double-blind, randomized, controlled clinical trial of caffeine versus placebo underwent actigraphy, polysomnography, and parental sleep questionnaires. Coprimary outcomes were total sleep time on actigraphy and apnea-hypopnea index on polysomnography. There were no significant differences in primary outcomes between the caffeine group and the placebo (adjusted mean difference of -6.7 [95% confidence interval (CI) = -15.3 to 2.0 min]; P = 0.13 for actigraphic total sleep time; and adjusted rate ratio [caffeine/placebo] for apnea-hypopnea index of 0.89 [95% CI = 0.55-1.43]; P = 0.63). Polysomnographic total recording time and total sleep time were longer in the caffeine group, but there was no difference in sleep efficiency between groups. The percentage of children with obstructive sleep apnea (8.2% of caffeine group versus 11.0% of placebo; P = 0.22) or elevated periodic limb movements of sleep (17.5% in caffeine group versus 11% in placebo group) was high, but did not differ significantly between groups. Therapeutic neonatal caffeine administration has no long-term effects on sleep duration or sleep apnea during childhood. Ex-preterm infants, regardless of caffeine status, are at risk for obstructive sleep apnea and periodic limb movements in later childhood.


Plasma Monocyte Chemotactic Protein-1 Is Associated With Acute Kidney Injury and Death After Cardiac Operations.
Monocyte chemotactic protein-1 (MCP-1; chemokine C-C ligand-2 [CCL-2]) is upregulated in ischemia-reperfusion injury and is a promising biomarker of inflammation in cardiac operations. We measured preoperative and postoperative plasma MCP-1 levels in adults undergoing cardiac operations to evaluate the association of perioperative MCP-1 levels with acute kidney injury (AKI) and death in Translational Research Investigating Biomarker Endpoints in AKI (TRIBE-AKI), a prospective, multicenter, observational cohort. Of the 972 participants in the study, AKI developed in 329 (34%), and severe AKI developed in 45 (5%). During a median follow-up of 2.9 years (interquartile range, 2.2 to 3.5 years), 119 participants (12%) died. MCP-1 levels were significantly higher in those who developed AKI and died than in those without AKI and death. Participants with a preoperative MCP-1 level in the highest tertile (>196 pg/mL) had an increased AKI risk than those in the lowest tertile (<147 pg/mL; odds ratio [OR], 1.43l; 95% confidence interval [CI], 1.00 to 2.05). The association appeared similar but was not significant for the severe AKI outcome (OR, 1.48; 95% CI, 0.62 to 3.54). Compared with participants with preoperative MCP-1 level in the lowest tertile, those in the highest tertile had higher adjusted risk of death (hazard ratio, 1.82; 95% CI, 1.40 to 2.38). Similarly, participants in the highest tertile had a higher adjusted risk of death (hazard ratio, 1.95; 95% CI, 1.09-3.49) than those with a postoperative MCP-1 level in the lowest tertile. Higher plasma MCP-1 is associated with increased AKI and risk of death after cardiac operations. MCP-1 could be used as a biomarker to identify high-risk patients for potential AKI prevention strategies in the setting of cardiac operations.


Vibrational properties of vitreous GeSe2 with the Becke-Lee-Yang-Parr density functional.
The vibrational properties of vitreous GeSe(2) are studied within the Becke-Lee-Yang-Parr (BLYP) generalized gradient approximation to density functional theory. For this purpose, we consider two models. The first one is derived from a chemically ordered network, previously generated through classical molecular dynamics. The second one is generated through first-principles molecular dynamics simulations based on the BLYP functional and shows a larger variety of structural motifs. We describe the structural and electronic properties of our models, including bond lengths, first-neighbor coordinations, and the electronic density of states. The vibrational properties are addressed through the vibrational density of states, the infrared spectrum, and the Raman spectrum. For the chemically ordered model, the vibrational properties obtained with the BLYP functional are of comparable quality with respect to previous results obtained with the Perdew-Wang (PW) functional. However, a global consideration of our results indicates that the BLYP functional yields an overall improved description for the vibrational properties of vitreous GeSe(2). The BLYP dielectric constants are closer to experimental values. Furthermore, we found that the Raman spectrum of the model generated through first-principles molecular dynamics within the BLYP scheme is largely improved with respect to an analogous model generated within the PW scheme.


The CBF1-independent Notch1 signal pathway activates human c-myc expression partially via transcription factor YY1.
Transcription factor Ying Yang 1 (YY1) indirectly regulates the C promoter-binding factor 1 (CBF1)-dependent Notch1 signaling via direct interaction with the Notch1 receptor intracellular domain (N1IC) on CBF1-response elements. To evaluate the possibility that the N1IC might modulate the gene expression of YY1 target genes through associating with YY1 on the YY1-response elements, we herein investigated the effect of Notch1 signaling on the expression of YY1 target genes. We found that the N1IC bound to the double-stranded oligonucleotides of YY1-response element to activate luciferase activity of the reporter gene with YY1-response elements through a CBF1-independent manner. Furthermore, the N1IC also bound to the promoter of human c-myc oncogene, a YY1 target gene, to elevate c-myc expression via a CBF1-independent pathway. The activation of reporter genes with YY1-response elements or human c-myc promoter by N1IC depended on the formation of N1IC-YY1-associated complex. To delineate the role of the Notch signal pathway in tumorigenesis, K562 cell lines expressing the N1IC were established. Compared with control cells, the proliferation and the tumor growth of N1IC-expressing K562 cells were suppressed. Taken together, these results suggest that the N1IC enhances the human c-myc promoter activity that is partially modulated by YY1 through a CBF1-independent pathway. However, the enhancement of c-myc expression by N1IC is insufficient to promote the tumor growth of K562 cells.


Spatial inference without a cognitive map: the role of higher-order path integration.
The cognitive map has been taken as the standard model for how agents infer the most efficient route to a goal location. Alternatively, path integration - maintaining a homing vector during navigation - constitutes a primitive and presumably less-flexible strategy than cognitive mapping because path integration relies primarily on vestibular stimuli and pace counting. The historical debate as to whether complex spatial navigation is ruled by associative learning or cognitive map mechanisms has been challenged by experimental difficulties in successfully neutralizing path integration. To our knowledge, there are only three studies that have succeeded in resolving this issue, all showing clear evidence of novel route taking, a behaviour outside the scope of traditional associative learning accounts. Nevertheless, there is no mechanistic explanation as to how animals perform novel route taking. We propose here a new model of spatial learning that combines path integration with higher-order associative learning, and demonstrate how it can account for novel route taking without a cognitive map, thus resolving this long-standing debate. We show how our higher-order path integration (HOPI) model can explain spatial inferences, such as novel detours and shortcuts. Our analysis suggests that a phylogenetically ancient, vector-based navigational strategy utilizing associative processes is powerful enough to support complex spatial inferences.


DDT ANTAGONISM TO DIELDRIN STORAGE IN ADIPOSE TISSUE OF RATS.
Storage of dieldrin in the adipose tissue of female rats was markedly depressed when DDT and dieldrin were fed simultaneously. The amount of dieldrin present in the tissues of rats fed 1 and 10 parts of dieldrin per million was significantly reduced by the addition of 5 ppm DDT to the feed. The addition of 50 ppm DDT to the feed caused a 15-fold reduction in the amount of dieldrin stored in rats fed 1 ppm dieldrin, and a 6-fold reduction in rats fed 10 ppm dieldrin. This antagonistic effect of DDT suggests that the criteria used in predicting the pharmacological effects of combined residues of related insecticides need some revision.


Risk factors for death among children aged 5-14 years hospitalised with pneumonia: a retrospective cohort study in Kenya.
There were almost 1 million deaths in children aged between 5 and 14 years in 2017, and pneumonia accounted for 11%. However, there are no validated guidelines for pneumonia management in older children and data to support their development are limited. We sought to understand risk factors for mortality among children aged 5-14 years hospitalised with pneumonia in district-level health facilities in Kenya. We did a retrospective cohort study using data collected from an established clinical information network of 13 hospitals. We reviewed records for children aged 5-14 years admitted with pneumonia between 1 March 2014 and 28 February 2018. Individual clinical signs were examined for association with inpatient mortality using logistic regression. We used existing WHO criteria (intended for under 5s) to define levels of severity and examined their performance in identifying those at increased risk of death. 1832 children were diagnosed with pneumonia and 145 (7.9%) died. Severe pallor was strongly associated with mortality (adjusted OR (aOR) 8.06, 95% CI 4.72 to 13.75) as were reduced consciousness, mild/moderate pallor, central cyanosis and older age (>9 years) (aOR >2). Comorbidities HIV and severe acute malnutrition were also associated with death (aOR 2.31, 95% CI 1.39 to 3.84 and aOR 1.89, 95% CI 1.12 to 3.21, respectively). The presence of clinical characteristics used by WHO to define severe pneumonia was associated with death in univariate analysis (OR 2.69). However, this combination of clinical characteristics was poor in discriminating those at risk of death (sensitivity: 0.56, specificity: 0.68, and area under the curve: 0.62). Children >5 years have high inpatient pneumonia mortality. These findings also suggest that the WHO criteria for classification of severity for children under 5 years do not appear to be a valid tool for risk assessment in this older age group, indicating the urgent need for evidence-based clinical guidelines for this neglected population.


Detailed mapping of the nuclear export signal in the Rous sarcoma virus Gag protein.
The Rous sarcoma virus (RSV) Gag polyprotein undergoes transient nuclear trafficking as an intrinsic part of the virus assembly pathway. Nuclear export of Gag is crucial for the efficient production of viral particles and is accomplished through the action of a leptomycin B (LMB)-dependent nuclear export signal (NES) in the p10 domain (L. Z. Scheifele, R. A. Garbitt, J. D. Rhoads, and L. J. Parent, Proc. Natl. Acad. Sci. USA 99:3944-3949, 2002). We have now mapped the nuclear export activity to the C-terminal portion of the p10 sequence and identified the four hydrophobic amino acids within this region that comprise a leucine-rich NES. Alteration of these hydrophobic residues resulted in the accumulation of Gag proteins within the nucleus and a budding defect greater than that obtained with LMB treatment of cells expressing the wild-type Gag protein (Scheifele et al., Proc. Natl. Acad. Sci. USA 99:3944-3949, 2002). In addition, export of Gag from the nucleus was found to be a rate-limiting step in virus-like particle production. Consistent with a role for the NES sequence in viral replication, this cluster of hydrophobic residues in p10 is conserved across a wide range of avian retroviruses. Furthermore, naturally occurring substitutions within this region in related viruses maintained nuclear export activity and remained sensitive to the activity of LMB. Using gain-of-function approaches, we found that the hydrophobic motif in p10 was sufficient to promote the nuclear export of a heterologous protein and was positionally independent within the Gag polyprotein. Finally, the export pathway was further defined by the ability of specific nucleoporin inhibitors to prevent the egress of Gag from the nucleus, thereby identifying additional cellular mediators of RSV replication.


Hostility predicts magnitude and duration of blood pressure response to anger.
The hypothesis that hostile and nonhostile individuals would differ in both magnitude and duration of cardiovascular reactivity to relived anger was tested. Participants were 66 older adults (mean age, 62; 38 women and 28 men; 70% Caucasian American, 30% African American). Each took part in a structured interview scored using the Interpersonal Hostility Assessment Technique. Later each relived a self-chosen anger memory while heart rate and systolic and diastolic blood pressures were measured continuously using an Ohmeda Finapres monitor. Hostile participants had larger and longer-lasting blood pressure responses to anger. African Americans also showed longer-lasting blood pressure reactivity to anger. Health and measurement implications are discussed.


Optimized anti-biofouling performance of bactericides/cellulose nanocrystals composites modified PVDF ultrafiltration membrane for micro-polluted source water purification.
The covalently functionalized cellulose nanocrystal (CNC) composites were synthesized by bonding common bactericides, such as dodecyl dimethyl benzyl ammonium chloride (DDBAC), ZnO and graphene oxide (GO) nanosheets, onto the CNC's surface. Then, the DDBAC/CNC, ZnO/CNC and GO/CNC nanocomposites modified polyvinylidene fluoride (PVDF) ultrafiltration membranes were fabricated by a simple one-step non-solvent induced phase separation (NIPS) process. The resultant hybrid membranes possessed porous and rough surfaces with more finger-like macropores that even extended through the entire cross-section. The hydrophilicity, permeability, antibacterial and antifouling performance and mechanism of the hybrid ultrafiltration membranes were evaluated and compared in detail, aiming at screening a superior hybrid membrane for practical application in micro-polluted source water purification. Among these newly-developed hybrid membranes, GO/CNC/PVDF exhibited an enhanced perm-selectivity with a water flux of 230 L/(m2 h bar) and humic acid rejection of 92%, the improved antibacterial activity (bacteriostasis rate of 93%) and antifouling performance (flux recovery rate (FRR) of >90%) being due to the optimized pore structure, higher surface roughness, incremental hydrophilicity and electronegativity. A lower biofouling level after three weeks' filtration of the actual micro-polluted source water further demonstrated that embedding the hydrophilic and antibacterial GO/CNC nanocomposite into the polymer matrix is an effective strategy to improve membrane anti-biofouling ability.


Acetazolamide for electrical status epilepticus in slow-wave sleep.
Electrical status epilepticus in slow-wave sleep (ESES) is characterized by nearly continuous spike-wave discharges during non-rapid eye movement (REM) sleep. ESES is present in Landau-Kleffner syndrome (LKS) and continuous spike and wave in slow-wave sleep (CSWS). Sulthiame has demonstrated reduction in spike-wave index (SWI) in ESES, but is not available in the United States. Acetazolamide (AZM) is readily available and has similar pharmacologic properties. Our aims were to assess the effect of AZM on SWI and clinical response in children with LKS and CSWS. Children with LKS or CSWS treated with AZM at our institution were identified retrospectively. Pre- and posttherapy electroencephalography (EEG) studies were evaluated for SWI. Parental and teacher report of clinical improvement was recorded. Six children met criteria for inclusion. Three children (50%) demonstrated complete resolution or SWI <5% after AZM. All children had improvement in clinical seizures and subjective improvement in communication skills and school performance. Five of six children had subjective improvement in hyperactivity and attention. AZM is a potentially effective therapy for children with LKS and CSWS. This study lends to the knowledge of potential therapies that can be used for these disorders, which can be challenging for families and providers.


Expression profiling of selected glutathione transferase genes in Zea mays (L.) seedlings infested with cereal aphids.
The purpose of this report was to evaluate the expression patterns of selected glutathione transferase genes (gst1, gst18, gst23 and gst24) in the tissues of two maize (Zea mays L.) varieties (relatively resistant Ambrozja and susceptible Tasty Sweet) that were colonized with oligophagous bird cherry-oat aphid (Rhopalosiphum padi L.) or monophagous grain aphid (Sitobion avenae L.). Simultaneously, insect-triggered generation of superoxide anion radicals (O2•-) in infested Z. mays plants was monitored. Quantified parameters were measured at 1, 2, 4, 8, 24, 48 and 72 h post-initial aphid infestation (hpi) in relation to the non-infested control seedlings. Significant increases in gst transcript amounts were recorded in aphid-stressed plants in comparison to the control seedlings. Maximal enhancement in the expression of the gst genes in aphid-attacked maize plants was found at 8 hpi (gst23) or 24 hpi (gst1, gst18 and gst24) compared to the control. Investigated Z. mays cultivars formed excessive superoxide anion radicals in response to insect treatments, and the highest overproduction of O2•- was noted 4 or 8 h after infestation, depending on the aphid treatment and maize genotype. Importantly, the Ambrozja variety could be characterized as having more profound increments in the levels of gst transcript abundance and O2•- generation in comparison with the Tasty Sweet genotype.


Stressful life events, genetic liability, and onset of an episode of major depression in women.
This study was undertaken to clarify how genetic liability and stressful life events interact in the etiology of major depression. Information about stressful life events and onset of major depressive episodes in the past year was collected in a population-based sample of female-female twin pairs including 2,164 individuals, 53,215 person-months of observation, and 492 onsets of depression. Nine "personal" and three aggregate "network" stressful events significantly predicted onset of major depression in the month of occurrence, four of which predicted onset with an odds ratio of > 10 and were termed "severe": death of a close relative, assault, serious marital problems, and divorce/breakup. Genetic liability also had a significant impact on risk of onset of depression. For severe stressful events, as well as for 10 of the 12 individual stressful events, the best-fitting model for the joint effect of stressful events and genetic liability on onset of major depression suggested genetic control of sensitivity to the depression-inducing effects of stressful life events. In individuals at lowest genetic risk (monozygotic twin, co-twin unaffected), the probability of onset of major depression per month was predicted to be 0.5% and 6.2%, respectively, for those unexposed and exposed to a severe event. In those at highest genetic risk (monozygotic twin, co-twin affected), these probabilities were 1.1% and 14.6%, respectively. Linear regression analysis indicated significant Genotype by Environment interaction in the prediction of onset of major depression. Genetic factors influence the risk of onset of major depression in part by altering the sensitivity of individuals to the depression-inducing effect of stressful life events.


Early specific cognitive-behavioural psychotherapy in subjects at high risk for bipolar disorders: study protocol for a randomised controlled trial.
Bipolar disorders (BD) are among the most severe mental disorders with first clinical signs and symptoms frequently appearing in adolescence and early adulthood. The long latency in clinical diagnosis (and subsequent adequate treatment) adversely affects the course of disease, effectiveness of interventions and health-related quality of life, and increases the economic burden of BD. Despite uncertainties about risk constellations and symptomatology in the early stages of potentially developing BD, many adolescents and young adults seek help, and most of them suffer substantially from symptoms already leading to impairments in psychosocial functioning in school, training, at work and in their social relationships. We aimed to identify subjects at risk of developing BD and investigate the efficacy and safety of early specific cognitive-behavioural psychotherapy (CBT) in this subpopulation. EarlyCBT is a randomised controlled multi-centre clinical trial to evaluate the efficacy and safety of early specific CBT, including stress management and problem solving strategies, with elements of mindfulness-based therapy (MBT) versus unstructured group meetings for 14 weeks each and follow-up until week 78. Participants are recruited at seven university hospitals throughout Germany, which provide in- and outpatient care (including early recognition centres) for psychiatric patients. Subjects at high risk must be 15 to 30 years old and meet the combination of specified affective symptomatology, reduction of psychosocial functioning, and family history for (schizo)affective disorders. Primary efficacy endpoints are differences in psychosocial functioning and defined affective symptomatology at 14 weeks between groups. Secondary endpoints include the above mentioned endpoints at 7, 24, 52 and 78 weeks and the change within groups compared to baseline; perception of, reaction to and coping with stress; and conversion to full BD. To our knowledge, this is the first study to evaluate early specific CBT in subjects at high risk for BD. Structured diagnostic interviews are used to map the risk status and development of disease. With our study, the level of evidence for the treatment of those young patients will be significantly raised. WHO International Clinical Trials Platform (ICTRP), identifier: DRKS00000444, date of registration: 16 June 2010.


Effect of disinfection of drinking water with ozone or chlorine dioxide on survival of Cryptosporidium parvum oocysts.
Demineralized water was seeded with controlled numbers of oocysts of Cryptosporidium parvum purified from fresh calf feces and subjected to different treatments with ozone or chlorine dioxide. The disinfectants were neutralized by sodium thiosulfate, and neonatal mice were inoculated intragastrically and sacrificed 7 days later for enumeration of oocyst production. Preliminary trials indicated that a minimum infection level of 1,000 oocysts (0.1-ml inoculum) per mouse was necessary to induce 100% infection. Treatment of water containing 10(4) oocysts per ml with 1.11 mg of ozone per liter (concentration at time zero [C0]) for 6 min totally eliminated the infectivity of the oocysts for neonatal mice. A level of 2.27 mg of ozone per liter (C0) was necessary to inactivate water containing 5 x 10(5) oocysts per ml within 8 min. Also, 0.4 mg of chlorine dioxide per liter (C0) significantly reduced infectivity within 15 min of contact, although some oocysts remained viable.


[Oral infection control in hospitalized patients: an approach to cardiologist and intensive care units doctors].
This paper aims to find the current level of periodontal med-care knowledge, as well as the possible existence of some oral infection control protocol regarding hospitalized patients. Our sample gathered 110 cardiologists and intensive care units doctors selected from medical teams of five Rio de Janeiro hospitals. Preliminary numbers: 75.4% said to have heard something about Periodontal Medicine, although only 30% out of this group admitted to have read something concerning such subject. On the other side, only 2.7% of the sample informed to do consistent information searching along their patients anamnese, while 58.2% out of this group admitted such procedure conditional to the patient's general state at the due moment. Through such numbers, we tend to come up to the conclusion that, be it either through direct or indirect Periodontal Medicine technical information (and consequently with regards to the absolut importance of preservation and control of oral biofilm and its impact on one's systemic health), the matter has been dimly spread among medical groups. The search also revealed the probability that Rio de Janeiro hospitals lack either units or agents designed for prevention and control of oral infection; consequently, such organizations do not have any kind of protocols, reliable or not, concerned to oral infections.


Effect of cognitive activity level on duration of post-concussion symptoms.
To determine the effect of cognitive activity level on duration of post-concussion symptoms. We conducted a prospective cohort study of patients who presented to a Sports Concussion Clinic within 3 weeks of injury between October 2009 and July 2011. At each visit, patients completed a scale that recorded their average level of cognitive activity since the previous visit. The product of cognitive activity level and days between visits (cognitive activity-days) was calculated and divided into quartiles. Kaplan-Meier Product Limit method was used to generate curves of symptom duration based on cognitive activity level. To adjust for other possible predictors of concussion recovery, we constructed a Cox proportional hazard model with cognitive activity-days as the main predictor. Of the 335 patients included in the study, 62% were male, 19% reported a loss of consciousness, and 37% reported experiencing amnesia at the time of injury. The mean age of participants was 15 years (range, 8-23) and the mean number of previous concussions was 0.76; 39% of athletes had sustained a previous concussion. The mean Post-Concussion Symptom Scale score at the initial visit was 30 (SD, 26). The overall mean duration of symptoms was 43 days (SD, 53). Of all variables assessed, only total symptom burden at initial visit and cognitive activity level were independently associated with duration of symptoms. Increased cognitive activity is associated with longer recovery from concussion. This study supports the use of cognitive rest and adds to the current consensus opinion.


Correlates of Vaginal Colonization with Group B Streptococci among Pregnant Women.
A study of genital colonization by group B streptococcus (GBS) was conducted in pregnant women in their third trimester, which is a known risk factor of morbidity and mortality among newborns. The present study was undertaken to study the prevalence and the correlates of vaginal colonization by GBS among pregnant women. This observational cross-sectional study was conducted during September 2002 to March 2004 on 524 pregnant women. Three high vaginal swabs were obtained from all the pregnant women admitted at term and in preterm labor. Two swabs were used for aerobic culture and the third one for gram staining. The first set of swabs was cultured on 5% Sheep blood agar plates. The second set of swabs were inoculated into Todd-Hewitt broth and then subcultured in 5% Sheep blood agar plates. The main outcome measures were the presence of GBS infection in comparison to the age group, gravida, gestational age, premature rupture of membrane (PROM), preterm labor and association with febrile spells of the present pregnancy. The culture positivity rate of GBS was 4.77% and coexistent organisms isolated were Candida species (36%), Staphylococcus aureus (8%) and Enterococcus species (8%). Culture positivity in the age group of 18-25 years was 5.71%, of which 5.74% were in their first pregnancy. The correlation between age group and gravida with GBS culture positivity was statistically insignificant. The culture positivity in <36 weeks of gestational age was 6.93%. This relation was statistically significant. Twenty-eight percent developed PROM. Sixty-four percent of culture positives had preterm labor. GBS infection among pregnant women was significantly correlated with the gestational age, PROM and preterm labor. In pregnancy GBS colonization causes asymptomatic bacteriuria or UTI. It is a well known cause of puerperal infections with amnionitis, endometritis and sepsis being the most commonly reported infections.[5].


Modeling the mechanism of CLN025 beta-hairpin formation.
Beta-hairpins are substructures found in proteins that can lend insight into more complex systems. Furthermore, the folding of beta-hairpins is a valuable test case for benchmarking experimental and theoretical methods. Here, we simulate the folding of CLN025, a miniprotein with a beta-hairpin structure, at its experimental melting temperature using a range of state-of-the-art protein force fields. We construct Markov state models in order to examine the thermodynamics, kinetics, mechanism, and rate-determining step of folding. Mechanistically, we find the folding process is rate-limited by the formation of the turn region hydrogen bonds, which occurs following the downhill hydrophobic collapse of the extended denatured protein. These results are presented in the context of established and contradictory theories of the beta-hairpin folding process. Furthermore, our analysis suggests that the AMBER-FB15 force field, at this temperature, best describes the characteristics of the full experimental CLN025 conformational ensemble, while the AMBER ff99SB-ILDN and CHARMM22* force fields display a tendency to overstabilize the native state.


Health worker experiences of and movement between public and private not-for-profit sectors-findings from post-conflict Northern Uganda.
Northern Uganda suffered 20 years of conflict which devastated lives and the health system. Since 2006, there has been investment in reconstruction, which includes efforts to rebuild the health workforce. This article has two objectives: first, to understand health workers' experiences of working in public and private not-for-profit (PNFP) sectors during and after the conflict in Northern Uganda, and second, to understand the factors that influenced health workers' movement between public and PNFP sectors during and after the conflict. A life history approach was used with 26 health staff purposively selected from public and PNFP facilities in four districts of Northern Uganda. Staff with at least 10 years' experience were selected, which resulted in a sample which was largely female and mid-level. Two thirds were currently employed in the public sector and just over a third in the PNFP sector. A thematic data analysis was guided by the framework analysis approach, analysis framework stages and ATLAS.ti software version 7.0. Analysis reveals that most of the current staff were trained in the PNFP sector, which appears to offer higher quality training experiences. During the conflict period, the PNFP sector also functioned more effectively and was relatively better able to support its staff. However, since the end of the conflict, the public sector has been reconstructed and is now viewed as offering a better overall package for staff. Most reported movement has been in that direction, and many in the PNFP sector state intention to move to the public sector. While there is sectoral loyalty on both sides and some bonds created through training, the PNFP sector needs to become more competitive to retain staff so as to continue delivering services to deprived communities in Northern Uganda. There has been limited previous longitudinal analysis of how health staff perceive different sectors and why they move between them, particularly in conflict-affected contexts. This article adds to our understanding, particularly for mid-level cadres, and highlights the need to ensure balanced health labour market incentives which take into account not only the changing context but also needs at different points in individuals' life cycles and across all core service delivery sectors.


Bevacizumab Maintenance Versus No Maintenance During Chemotherapy-Free Intervals in Metastatic Colorectal Cancer: A Randomized Phase III Trial (PRODIGE 9).
Purpose Conflicting results are reported for maintenance treatment with bevacizumab during chemotherapy-free intervals (CFI) in metastatic colorectal cancer after induction chemotherapy. Patients and Methods In this open-label, phase III, randomized controlled trial, we compared the tumor control duration (TCD) observed with bevacizumab maintenance and with no treatment (observation) during CFI subsequent to induction chemotherapy with 12 cycles of fluorouracil, leucovorin, and irinotecan plus bevacizumab. After disease progression, the induction regimen was repeated for eight cycles, followed by a new CFI. Results From March 2010 to July 2013, 491 patients were randomly assigned. Disease progression or death occurred during induction chemotherapy in 85 patients (17%); 261 patients (53%) had at least one reinduction, 107 (22%) had two reinductions, and 56 (11%) had three or more reinductions. The median TCD was 15 months in both groups; the median progression-free survival (PFS) from randomization was 9.2 and 8.9 months in the maintenance group and observation groups, respectively. The TCD observed in both groups was higher compared with the TCD hypotheses of the trial. The median overall survival (OS) was 21.7 and 22.0 months in the maintenance and observation groups, respectively. In the per-protocol population, defined as patients with at least one reinduction after the first progression, the median duration of the first CFI was 4.3 months in both arms; the median TCD was 17.8 and 23.3 months ( P = .339), the median PFS was 9.9 and 9.5 months, and the median OS was 27.6 and 28.5 months in the maintenance and observation groups, respectively. Multivariable analysis revealed that female gender, WHO performance status ≥ 2, and unresected primary tumors were associated with a shorter TCD. Conclusion Bevacizumab maintenance monotherapy did not improve TCD, CFI duration, PFS, or OS.


A community-based aquatic exercise program to improve endurance and mobility in adults with mild to moderate intellectual disability.
The purpose of this study was to investigate the impact of a community-based aquatic exercise program on physical performance among adults with mild to moderate intellectual disability (ID). Twenty-two community-dwelling adults with mild to moderate ID volunteered to participate in this study. Participants completed an 8-week aquatic exercise program (2 days/wk, 1 hr/session). Measures of physical performance, which were assessed prior to and following the completion of the aquatic exercise program, included the timed-up-and-go test, 6-min walk test, 30-sec chair stand test, 10-m timed walk test, hand grip strength, and the static plank test. When comparing participants' measures of physical performance prior to and following the 8-week aquatic exercise program, improvements were seen in all measures, but the change in scores for the 6-min walk test, 30-sec chair stand test, and the static plank test achieved statistical significance (P<0.05). An 8-week group aquatic exercise program for adults with ID may promote improvements in endurance and balance/mobility.


Self-reported use of novel psychoactive substances among attendees of electronic dance music venues.
Novel psychoactive substances (NPSs) continue to emerge in the United States and worldwide. Few epidemiological studies have examined the prevalence and correlates of use. We examined the extent of NPS use in a high-risk population-attendees of electronic dance music (EDM) parties at nightclubs and festivals. We surveyed 682 adults (age 18-25) entering EDM events at nightclubs and festivals in New York City (NYC) in 2015. A variation of time-space sampling was used. We examined the prevalence of self-reported use of 196 NPS and correlates of any NPS use. Over a third (35.1%) of participants reported lifetime use of any NPS. Self-reported use of synthetic cannabinoids was most prevalent (16.3%), followed by psychedelic phenethylamines (14.7%; 2C series: 10.3%, 2-(4-iodo-2,5-dimethoxyphenyl)-N-[(2-methoxyphenyl)methyl]ethanamine [NBOMe] series: 9.0%, Dox series: 3.5%), synthetic cathinones ("bath salts", 6.9%), other psychedelics (6.6%), tryptamines (5.1%), and dissociatives (4.3%). 2C-I was the most prevalent 2C series drug (5.1%); methylone was the most prevalent synthetic cathinone (3.3%), 2-MeO-ketamine was the most prevalent dissociative (3.7%), and 1P-lysergic acid diethylamide (LSD) (2.9%) was the most prevalent non-phenethylamine psychedelic. Risk factors for NPS use included Ecstasy/MDMA/Molly, LSD, and ketamine use; identifying as bisexual (compared to heterosexual), reporting higher frequency of nightclub/festival attendance, and being surveyed outside of a festival (compared to those surveyed outside of nightclubs). NPS use is prevalent in the nightclub and festival scenes in NYC. Since individuals in these scenes-especially frequent attendees-are at high risk for use, prevention and harm reduction services need to be geared toward this population.


Emergent ultra-long-range interactions between active particles in hybrid active-inactive systems.
Particle-particle interactions determine the state of a system. Control over the range of such interactions as well as their magnitude has been an active area of research for decades due to the fundamental challenges it poses in science and technology. Very recently, effective interactions between active particles have gathered much attention as they can lead to out-of-equilibrium cooperative states such as flocking. Inspired by nature, where active living cells coexist with lifeless objects and structures, here we study the effective interactions that appear in systems composed of active and passive mixtures of colloids. Our systems are 2D colloidal monolayers composed primarily of passive (inactive) colloids, and a very small fraction of active (spinning) ferromagnetic colloids. We find an emergent ultra-long-range attractive interaction induced by the activity of the spinning particles and mediated by the elasticity of the passive medium. Interestingly, the appearance of such interaction depends on the spinning protocol and has a minimum actuation timescale below which no attraction is observed. Overall, these results clearly show that, in the presence of elastic components, active particles can interact across very long distances without any chemical modification of the environment. Such a mechanism might potentially be important for some biological systems and can be harnessed for newer developments in synthetic active soft materials.


Role of ASC in hypoxia-mediated cell death in pancreatic cancer.
ASC, an apoptosis-associated speck-like protein, can regulate apoptosis in response to various types of cell death stimuli. In this study, we investigated the role of ASC in hypoxia-mediated cell death in pancreatic cancer. ASC was inducible under a 1% O2 hypoxic condition in pancreatic cancer cells, which was HIF1α-dependent but p53-independent. Two representative chemotherapeutic agents for pancreatic cancer, 5-fluorouracil and gemcitabine, promoted cell death in a p53-dependent manner; however, 1% hypoxia caused chemoresistance to these drugs. Western blot analysis of this condition showed that expression of Bax, a pro-apoptotic gene, decreased, while the anti-apoptotic genes IAP-2 and survivin increased. These results suggest that, although hypoxia induces both pro-apoptotic and anti-apoptotic genes, the total balance seems to be anti-apoptotic dominant, which might explain chemoresistance in pancreatic cancer. To overcome this anti-apoptotic dominant condition, we infected adenovirus-expressing ASC into pancreatic cancer cells. The expressed ASC induced cell death even under 20% normoxia, and was enhanced by hypoxia. Our data demonstrate the possible mechanism of chemoresistance under hypoxia in pancreatic cancer cells, thereby suggesting the potential use of ASC as a new treatment strategy for pancreatic cancer.


Genome assembly forensics: finding the elusive mis-assembly.
We present the first collection of tools aimed at automated genome assembly validation. This work formalizes several mechanisms for detecting mis-assemblies, and describes their implementation in our automated validation pipeline, called amosvalidate. We demonstrate the application of our pipeline in both bacterial and eukaryotic genome assemblies, and highlight several assembly errors in both draft and finished genomes. The software described is compatible with common assembly formats and is released, open-source, at http://amos.sourceforge.net.


A prospective matched study on symptomatic dengue in pregnancy.
Dengue fever is an increasing problem worldwide, but consequences during pregnancy remain unclear. Much of the available literature suffers from methodological biases that compromise the validity of clinical recommendations. We conducted a matched cohort study during an epidemic in French Guiana to compare events and pregnancy outcomes between two paired groups of pregnant women: women having presented with symptomatic dengue during pregnancy (n = 73) and women having had neither fever nor dengue during pregnancy (n = 219). Women in each arm were matched by place of follow up, gestation weeks at inclusion, and place of residence. Dengue infection was considered to be confirmed if viral RNA, N S1 antigen, the seroconversion of IgM antibodies or the presence of IgM was detected in collected samples. According to the 2009 WHO classification, 27% of the women with symptomatic dengue had at least one clinical or biological warning sign. These complications occurred after the 28th week of gestation in 55% of cases. The medical history, socioeconomic status and demographic characteristics were included in multivariate analysis. Exposure to dengue during pregnancy was not significantly associated with prematurity, small for gestational age infants, hypertension or emergency caesarian section. Maternal dengue with warning signs was a risk factor for peripartum hemorrhage with adjusted relative risk = 8.6(95% CI = 1.2-62). There was a near significant association between dengue and in utero death (p = 0.09). This prospective comparative study underlined the importance of taking into account potential confounders between exposure to dengue and the occurrence of obstetrical events. It also confirms the need for increased vigilance for pregnant women with dengue, particularly for women who present with severe dengue.


Suggestive evidence for linkage to chromosome 13qter for autosomal dominant type 1 porencephaly.
A large three-generation family with autosomal dominant type 1 porencephaly from southern Italy was studied. A high rate of miscarriages was observed. Of the nine affected individuals, four displayed a severe phenotype, and five had slight pyramidal signs or mild cognitive abnormalities. The MRI study disclosed unilateral porencephalic cyst, or colpocephaly. A genome-wide screen resulted in suggestive evidence for linkage to chromosome 13qter with a maximum logarithm-of-the-odds score of 3.16, from multipoint analysis, with marker D13S285.


Preferences for cancer investigation: a vignette-based study of primary-care attendees.
The UK lags behind many European countries in terms of cancer survival. Initiatives to address this disparity have focused on barriers to presentation, symptom recognition, and referral for specialist investigation. Selection of patients for further investigation has come under particular scrutiny, although preferences for referral thresholds in the UK population have not been studied. We investigated preferences for diagnostic testing for colorectal, lung, and pancreatic cancers in primary-care attendees. In a vignette-based study, researchers recruited individuals aged at least 40 years attending 26 general practices in three areas of England between Dec 6, 2011, and Aug 1, 2012. Participants completed up to three of 12 vignettes (four for each of lung, pancreatic, and colorectal cancers), which were randomly assigned. The vignettes outlined a set of symptoms, the risk that these symptoms might indicate cancer (1%, 2%, 5%, or 10%), the relevant testing process, probable treatment, possible alternative diagnoses, and prognosis if cancer were identified. Participants were asked whether they would opt for diagnostic testing on the basis of the information in the vignette. 3469 participants completed 6930 vignettes. 3052 individuals (88%) opted for investigation in their first vignette. We recorded no strong evidence that participants were more likely to opt for investigation with a 1% increase in risk of cancer (odds ratio [OR] 1·02, 95% CI 0·99-1·06; p=0·189), although the association between risk and opting for investigation was strong when colorectal cancer was analysed alone (1·08, 1·03-1·13; p=0·0001). In multivariable analysis, age had an effect in all three cancer models: participants aged 60-69 years were significantly more likely to opt for investigation than were those aged 40-59 years, and those aged 70 years or older were less likely. Other variables associated with increased likelihood of opting for investigation were shorter travel times to testing centre (colorectal and lung cancers), a family history of cancer (colorectal and lung cancers), and higher household income (colorectal and pancreatic cancers). Participants in our sample expressed a clear preference for diagnostic testing at all risk levels, and individuals want to be tested at risk levels well below those stipulated by UK guidelines. This willingness should be considered during design of cancer pathways, particularly in primary care. The public engagement with our study should encourage general practitioners to involve patients in referral decision making. The National Institute for Health Research Programme Grants for Applied Research programme.


Practice size: impact on consultation length, workload, and patient assessment of care.
Variations in practice list size are known to be associated with changes in a number of markers of primary care. Few studies have addressed the issue of how single-handed and smaller practices compare with larger group practices and what might be the optimal size of a general practice. To examine variations in markers of the nature of the care being provided by practices of various size. Practice profile questionnaire survey. A randomised sample of general practitioners (GPs) and practices from two inner-London areas, stratified according to practice size and patients attending the practice over a two-week period. Average consultation length was calculated over 200 consecutive consultations. A patient survey using the General Practice Assessment Survey instrument was undertaken in each practice. A practice workload survey was carried out over a two-week period. These outcome measures were examined in relation to five measures of practice size based on total list size and the number of doctors providing care. Out of 202 pratices approached, 54 provided analysable datasets. The patient survey response rate was 7247/11,000 (66%). Smaller practices had shorter average consultation lengths and reduced practice performance scores compared with larger practices. The number of patients corrected for the number of doctors providing care was an important predictor of consultation length in group practices. Responders from smaller practices reported improved accessibility of care and receptionist performance, better continuity of care compared with larger practices, and no disadvantage in relation to 10 other dimensions of care. Practices with smaller numbers of patients per doctor had longer average consultation lengths than those with larger numbers of patients per doctor. Defining the optimal size of practice is a complex decision in which the views of doctors, patients, and health service managers may be at variance. Some markers of practice performance are related to the total number of patients cared for, but the practice size corrected for the number of available doctors gives a different perspective on the issue. An oversimplistic approach that fails to account for the views of patients as well as health professionals is likely to be disadvantageous to service planning.


Pathway from central obesity to childhood asthma. Physical fitness and sedentary time are leading factors.
Available prospective studies of obesity and asthma have used only body mass index (BMI) as an indicator for adiposity; studies using detailed obesity measures are lacking, and the role of physical fitness level and sedentary time remains unexplored in the link between obesity and asthma. To compare various anthropometric measures of obesity in relation to childhood asthma, and to further characterize the interrelations among central obesity, physical fitness level, sedentary time, and asthma. The nationwide Taiwan Children Health Study followed 2,758 schoolchildren from fourth to sixth grade, annually collecting data regarding physical fitness, sedentary time, obesity measures (comprising body weight and height, abdominal and hip circumference, skin fold thickness, and body composition), asthma, and pulmonary function tests. The generalized estimating equation was used for 3 years of repeated measurements to analyze the interrelation among obesity, sedentary time, physical fitness level, and asthma; a structural equation model was used to explore the pathogenesis among these factors. Asthma incidence was analyzed during a 2-year follow-up among centrally obese and nonobese groups in baseline children without asthma. Central obesity most accurately predicts asthma. Low physical fitness levels and high screen time increase the risk of central obesity, which leads to asthma development. Obesity-related reduction in pulmonary function is a possible mechanism in the pathway from central obesity to asthma. Central obesity measures should be incorporated in childhood asthma risk predictions. Children are encouraged to increase their physical fitness levels and reduce their sedentary time to prevent central obesity-related asthma.


Structures of the fungal dynamin-related protein Vps1 reveal a unique, open helical architecture.
Dynamin-related proteins (DRPs) are large multidomain GTPases required for diverse membrane-remodeling events. DRPs self-assemble into helical structures, but how these structures are tailored to their cellular targets remains unclear. We demonstrate that the fungal DRP Vps1 primarily localizes to and functions at the endosomal compartment. We present crystal structures of a Vps1 GTPase-bundle signaling element (BSE) fusion in different nucleotide states to capture GTP hydrolysis intermediates and concomitant conformational changes. Using cryoEM, we determined the structure of full-length GMPPCP-bound Vps1. The Vps1 helix is more open and flexible than that of dynamin. This is due to further opening of the BSEs away from the GTPase domains. A novel interface between adjacent GTPase domains forms in Vps1 instead of the contacts between the BSE and adjacent stalks and GTPase domains as seen in dynamin. Disruption of this interface abolishes Vps1 function in vivo. Hence, Vps1 exhibits a unique helical architecture, highlighting structural flexibilities of DRP self-assembly.


A wide-angle high Mach number modal expansion for infrasound propagation.
The use of modal expansions to solve the problem of atmospheric infrasound propagation is revisited. A different form of the associated modal equation is introduced, valid for wide-angle propagation in atmospheres with high Mach number flow. The modal equation can be formulated as a quadratic eigenvalue problem for which there are simple and efficient numerical implementations. A perturbation expansion for the treatment of attenuation, valid for stratified media with background flow, is derived as well. Comparisons are carried out between the proposed algorithm and a modal algorithm assuming an effective sound speed, including a real data case study. The comparisons show that the effective sound speed approximation overestimates the effect of horizontal wind on sound propagation, leading to errors in traveltime, propagation path, trace velocity, and absorption. The error is found to be dependent on propagation angle and Mach number.


Population Genomic Analysis Reveals Differential Evolutionary Histories and Patterns of Diversity across Subgenomes and Subpopulations of Brassica napus L.
The allotetraploid species Brassica napus L. is a global crop of major economic importance, providing canola oil (seed) and vegetables for human consumption and fodder and meal for livestock feed. Characterizing the genetic diversity present in the extant germplasm pool of B. napus is fundamental to better conserve, manage and utilize the genetic resources of this species. We used sequence-based genotyping to identify and genotype 30,881 SNPs in a diversity panel of 782 B. napus accessions, representing samples of winter and spring growth habits originating from 33 countries across Europe, Asia, and America. We detected strong population structure broadly concordant with growth habit and geography, and identified three major genetic groups: spring (SP), winter Europe (WE), and winter Asia (WA). Subpopulation-specific polymorphism patterns suggest enriched genetic diversity within the WA group and a smaller effective breeding population for the SP group compared to WE. Interestingly, the two subgenomes of B. napus appear to have different geographic origins, with phylogenetic analysis placing WE and WA as basal clades for the other subpopulations in the C and A subgenomes, respectively. Finally, we identified 16 genomic regions where the patterns of diversity differed markedly from the genome-wide average, several of which are suggestive of genomic inversions. The results obtained in this study constitute a valuable resource for worldwide breeding efforts and the genetic dissection and prediction of complex B. napus traits.


Automatic detection of anatomical landmarks in uterine cervix images.
The work focuses on a unique medical repository of digital cervicographic images ("Cervigrams") collected by the National Cancer Institute (NCI) in longitudinal multiyear studies. NCI, together with the National Library of Medicine (NLM), is developing a unique web-accessible database of the digitized cervix images to study the evolution of lesions related to cervical cancer. Tools are needed for automated analysis of the cervigram content to support cancer research. We present a multistage scheme for segmenting and labeling regions of anatomical interest within the cervigrams. In particular, we focus on the extraction of the cervix region and fine detection of the cervix boundary; specular reflection is eliminated as an important preprocessing step; in addition, the entrance to the endocervical canal (the "os"), is detected. Segmentation results are evaluated on three image sets of cervigrams that were manually labeled by NCI experts.


Research on the thermal relaxation effect of nonplanar four-frequency differential laser gyro.
The nonplanar four-frequency differential laser gyro (NFFDLG, also called ZLG in the United States) has many advantages, such as the absence of mechanical noise, a whole solid state, high stability of scale factor, and no time delay. Because of its excellent performance, NFFDLG is currently the subject of research. In this paper, the error induced by the thermal relaxation effect is investigated both theoretically and experimentally, and some methods and conclusions that could restrain this error effectively are proposed. The experiment data confirm reliably that NFFDLG has a thermal relaxation effect error during overturning. At the same time, it is pointed out that the error can be minimized by reducing the stop interval. Furthermore, since the sum frequency could be affected more seriously by the overturning thermal relaxation effect than the difference frequency could, the former one can be used as one more sensitive condition in actual applications. As far as we know, the overturning thermal relaxation effect was first investigated in China, which is even not mentioned publicly.


The glycolytic inhibitor 2-deoxy-D-glucose enhances the efficacy of etoposide in ehrlich ascites tumor-bearing mice.
Earlier studies have shown that 2-deoxy-D-glucose (2-DG), a glucose analogue and inhibitor of glycolytic ATP production significantly enhances the cytotoxic effects of anticancer agents like topoisomerase inhibitors (etoposide and camptothecin) and a radiomimetic drug (bleomycin) in established human tumor cell lines. Therefore, combination of 2-DG and DNA damage causing cytotoxic agents could be very useful in enhancing local tumor control. The purpose of the present studies was to investigate the therapeutic effects of etoposide and 2-DG in Ehrlich ascites tumor (EAT) bearing mice, grown as solid tumor as well as in the ascites form. Cell growth, cell cycle perturbations (flow cytometry), cytogenetic damage (micronuclei assay) and apoptosis (DNA content, morphological changes) were studied as parameters of cellular response, while delay in tumor growth and cure rate (tumor free survival) were evaluated as parameters of systemic response. Body weight and general condition as well as the damage to bone marrow and spleen was monitored to evaluate normal tissue toxicity. Intraperitoneal administration of etoposide (30 mg/Kg b. wt.) resulted in significant tumor growth delay and cure (approximately 11%) only in subcutaneous tumors leading to local tumor control. When etoposide was combined with 2-DG (2 g/Kg b. wt.; i.v./i.p.; 4 h after etoposide injection), these effects were further enhanced resulting in a cure rate of approximately 22% in case of subcutaneous tumors and 20% in ascites bearing mice. Analysis of cells obtained from ascitic fluid as well as solid tumors during follow up clearly showed that etoposide induced cell death was mainly due to apoptosis, which was enhanced further by 2-DG. Although, there was a significant level of toxicity revealed by reduced animal survival, decrease in body weight and damage to sensitive organ status like spleen and bone marrow at 60 mg/Kg b. wt. of etoposide, it was not significant at 30 mg/Kg b.wt. 2-DG, however, did not enhance the etoposide toxicity at both the doses. These results indicate that the administration of 2-DG can improve the local control of tumors without increasing normal tissue toxicity, thereby enhancing the therapeutic efficacy of topoisomerase inhibitor based anticancer drugs like etoposide.


[A global vision of adverse reactions to foods: food allergy and food intolerance].
Over the last years, there has been an increase in adverse food reactions, probably associated with life style changes in the past decades. An adverse food reaction is any clinically abnormal response that can be attributed to ingestion, contact or inhalation of a food, its derivatives or an additive contained in it. They can be classified as food allergy or intolerance. Food allergies are usually immune-mediated, associated with IgE. Adverse reactions to food have a large clinical and social repercussion, which can be fatal in some cases and impair the quality of life of patients. This implies directly the services of collective catering and food manufacturing, which is why a legislature and regulations were implemented for its correct management. The most allergenic foods change according to the age group; being the egg the most frequent in children under 5 years, and fresh fruits in the older than 5 years. The most frequent clinical manifestations are cutaneous-mucous type. Food intolerances may be due to a pharmacological, metabolic, mixed or idiosyncratic mechanism. Clinical manifestations are usually dose dependent. The most common and known food intolerance is lactose, which is a metabolic type. The main treatment of both types of adverse reaction to foods is avoidance of the causal food of the reaction.


Changes in Skinfold Thicknesses and Body Fat in Ultra-endurance Cyclists.
The present study investigated the changes in single skinfold thicknesses and body fat during an ultra-endurance cycling race. One hundred and nineteen ultra-endurance cyclists in the 'Swiss Cycling Marathon' covering a distance of 600 km were included. Changes in skinfold thickness, fat mass, skeletal muscle mass and total body water were estimated using anthropometric methods. The subjects were riding at a mean speed of 23.5±4.0 km/h and finished the race within 1,580±296 min. During the race, body mass decreased by 1.5±1.2 kg (P<0.001), and fat mass decreased by 1.5±1.1 kg (P<0.001). Skeletal muscle mass and total body water remained unchanged (P>0.05). The decrease in body mass correlated to the decrease in fat mass (r = 0.20, P=0.03). The skinfold thicknesses at pectoral (-14.7%), abdominal (-14.9%), and thigh (-10.2%) site showed the largest decrease. The decrease in abdominal skinfold was significantly and negatively related to cycling speed during the race (r = -0.31, P<0.001). Cycling 600 km at ∼23 km/h led to a decrease in fat mass and in all skinfold thicknesses. The largest decrease in skinfold thickness was recorded for pectoral, abdominal, and thigh site. The decrease in abdominal skinfold thickness was negatively related to cycling speed. The body seems to reduce adipose subcutaneous fat during an ultra-endurance performance at the site of the thickest skinfold.


Ethical considerations concerning computers in medicine in the 1980s.
The advent of information technology and computers in medicine two decades ago posed a new set of ethical problems. In recent years, these problems have been compounded by the increasing use of computers for supporting clinical decisions as well as administration and record keeping. This presentation considers some of the problems which are raised by the use of computers to support clinical decisions, under the various headings of beneficence, non-maleficence, autonomy and justice. For each aspect the problems raised are described and the implications discussed.


Evaluation of L-tryptophan for treatment of insomnia: a review.
Sleep laboratory and outpatient studies of the hypnotic efficacy of the amino acid L-tryptophan are reviewed, with particular emphasis on evaluation of therapeutic effectiveness in the treatment of insomnia. In younger situational insomniacs, whose sleep problem consists solely of longer than usual sleep latencies, L-tryptophan is effective in reducing sleep onset time on the first night of administration in doses ranging from 1 to 15 g. In more chronic, well-established sleep-onset insomnia or in more severe insomnias characterized by both sleep onset and sleep maintenance problems, repeated administration of low doses of L-tryptophan over time may be required for therapeutic improvement. In these patients, hypnotic effects appear late in the treatment period or, as shown in some studies, even after discontinuation of treatment. The improvement in sleep measures post-treatment has given rise to use of a treatment regimen known as "interval therapy", in which L-tryptophan treatment alternates with an L-tryptophan-free interval until improvement occurs. The absence of side effects and lack of development of tolerance in long-term use are important factors in the decision to embark upon a trial of L-tryptophan treatment. In addition, L-tryptophan administration is not associated with impairment of visuomotor, cognitive, or memory performance, nor does it elevate threshold for arousal from sleep.


Telomere-independent ageing in the longest-lived non-colonial animal, Arctica islandica.
The shortening of telomeres as a causative factor in ageing is a widely discussed hypothesis in ageing research. The study of telomere length and its regenerating enzyme telomerase in the longest-lived non-colonial animal on earth, Arctica islandica, should inform whether the maintenance of telomere length plays a role in reaching the extreme maximum lifespan (MLSP) of >500years in this species. Since longitudinal measurements on living animals cannot be achieved, a cross-sectional analysis of a short-lived (MLSP 40years from the Baltic Sea) and a long-lived population (MLSP 226years Northeast of Iceland) and in different tissues of young and old animals from the Irish Sea was performed. A high heterogeneity of telomere length was observed in investigated A. islandica over a wide age range (10-36years for the Baltic Sea, 11-194years for Irish Sea, 6-226years for Iceland). Constant telomerase activity and telomere lengths were detected at any age and in different tissues; neither correlated with age or population habitat. Stable telomere maintenance might contribute to the long lifespan of A. islandica. Telomere dynamics are no explanation for the distinct MLSPs of the examined populations and thus the cause of it remains to be investigated.


Mast cells and neutrophils mediate peripheral motor pathway degeneration in ALS.
Neuroinflammation is a recognized pathogenic mechanism underlying motor neuron degeneration in amyotrophic lateral sclerosis (ALS), but the inflammatory mechanisms influencing peripheral motor axon degeneration remain largely unknown. A recent report showed a pathogenic role for c-Kit-expressing mast cells mediating inflammation and neuromuscular junction denervation in muscles from SOD1G93A rats. Here, we have explored whether mast cells infiltrate skeletal muscles in autopsied muscles from ALS patients. We report that degranulating mast cells were abundant in the quadriceps muscles from ALS subjects but not in controls. Mast cells were associated with myofibers and motor endplates and, remarkably, interacted with neutrophils forming large extracellular traps. Mast cells and neutrophils were also abundant around motor axons in the extensor digitorum longus muscle, sciatic nerve, and ventral roots of symptomatic SOD1G93A rats, indicating that immune cell infiltration extends along the entire peripheral motor pathway. Postparalysis treatment of SOD1G93A rats with the tyrosine kinase inhibitor drug masitinib prevented mast cell and neutrophil infiltration, axonal pathology, secondary demyelination, and the loss of type 2B myofibers, compared with vehicle-treated rats. These findings provide further evidence for a yet unrecognized contribution of immune cells in peripheral motor pathway degeneration that can be therapeutically targeted by tyrosine kinase inhibitors.


Epitaxial relationships in urolithiasis: the brushite-whewellite system.
1. Whewellite (calcium oxalate monohydrate) crystals were found to induce epitaxially the heterogeneous nucleation of brushite (calcium monohydrogen phosphate dihydrate) from its metastable supersaturated solution in approximately one-quarter of the time required for spontaneous precipitation in the absence of added nucleating agents. Scanning electron-microscope observation of the crystalline phase showed brushite crystals originating from the whewellite seed crystals. 2. Crystal growth, upon nucleation, proceeded rapidly, and the metastable solutions quickly approached saturation. 3. Brushite crystals also induced the precipitation of calcium oxalate crystals in about one-quarter of the time required for spontaneous precipitation; however, the rate of crystal growth was considerably slower. In support of the chemical data, scanning electron micrographs showed few crystals of calcium oxalate nucleated on the surface of the brushite seed. 4. The results provide some insight into the cause of stones containing calcium oxalate or calcium phosphate (or both), which form in the normally acid environment of human urine.


Expression and disposition of the murine mammary tumor virus (MuMTV) envelope gene products by murine mammary tumor cells.
Three murine mammary tumor virus (MuMTV)-producing epithelial cell lines derived from murine mammary tumors were examined in order to identify the MuMTV-specific cell surface antigens and their distribution on the cell surface, to study the kinetics of the MuMTV envelope precursor processing, virus assembly, and release, and to characterize the soluble MuMTV antigens that are shed into culture medium. Cell surface labeling experiments showed that only the mature MuMTV envelope glycoproteins gp52 and gp36 were exposed on the cell surface, and that gp52 was more abundant than gp36. In cells producing large quantities of MuMTV, expression of gp52 on the cell surface was shown by immunoelectron microscopy to be localized predominantly on the surface of budding virions and not on smooth areas of the cell surface where virus was not budding. The cell surface associated gp36 was found not to be incorporated into budding virions. A few cells in all three cell lines were found to produce only a few or no MuMTV particles and in these cells, unlike in the high virus-producing cells, considerable quantities of gp52 were expressed on the surface membrane. All three cell lines were found to shed large amounts of the MuMTV env precursor polyprotein as well as the mature non-virion-associated glycoprotein, gp52, into the culture medium. The envelope precursor protein (P75env) that was shed into the culture medium was found to differ from the predominant form of the cellular env precursor (Pr70env) in that (1) P75env migrated with an apparent higher molecular weight than Pr70env in SDS gels; (2) Pr70env contained only the core oligosaccharide, whereas P75env contained fucose in addition to the core sugars; (3) two-dimensional gel electrophoretic analysis showed that Pr70env could be resolved into three to four components migrating in the basic region of the isoelectric focussing gel (pH 7-8), whereas P75env was resolved into 9-13 components migrating in a more acidic region of the gel (pH 5-7). The molecular structure of the exfoliated gp52 was found to be similar to that of the gp52 that was incorporated into the virions although the virion-associated gp52 was not the source of the gp52 in the medium. Our quantitative pulse-chase studies suggest that of the two populations of MuMTV env precursors that are present in MuMTV-producing cells, only Pr70env is processed intracellularly to give rise to the mature MuMTV envelope proteins gp52 and gp36.


Mechanistic insights into dopaminergic and serotonergic neurotransmission - concerted interactions with helices 5 and 6 drive the functional outcome.
Brain functions rely on neurotransmitters that mediate communication between billions of neurons. Disruption of this communication can result in a plethora of psychiatric and neurological disorders. In this work, we combine molecular dynamics simulations, live-cell biosensor and electrophysiological assays to investigate the action of the neurotransmitter dopamine at the dopaminergic D2 receptor (D2R). The study of dopamine and closely related chemical probes reveals how neurotransmitter binding translates into the activation of distinct subsets of D2R effectors (i.e.: Gi2, GoB, Gz and β-arrestin 2). Ligand interactions with key residues in TM5 (S5.42) and TM6 (H6.55) in the D2R binding pocket yield a dopamine-like coupling signature, whereas exclusive TM5 interaction is typically linked to preferential G protein coupling (in particular GoB) over β-arrestin. Further experiments for serotonin receptors indicate that the reported molecular mechanism is shared by other monoaminergic neurotransmitter receptors. Ultimately, our study highlights how sequence variation in position 6.55 is used by nature to fine-tune β-arrestin recruitment and in turn receptor signaling and internalization of neurotransmitter receptors.


Pentoxifylline as adjunctive therapy in leg ulcer management.
Refractory or recurring leg ulcers typically signify the presence of impaired venous or arterial microcirculation (or both). A current hypothesis suggests that local oxygen and nutrient deficits occur because capillary lumens narrow and become permeable to fibrinogen and proteins by activated white blood cells and inadequate fibrinolysis. As a result, the deposition of a relatively impermeable perivascular fibrin sheath occurs, preventing adequate delivery of oxygen and nutrients. Therapy should therefore aim at reversing these deficits and improving wound healing. Pentoxifylline (Trental), used in combination with local wound care and appropriate antibiotic therapy, has been successful in significantly improving the healing of refractory leg ulcers. Pentoxifylline can reverse the abnormal function of white and red blood cells and platelets, as well as reduce blood viscosity and vascular permeability. The mechanisms of action of pentoxifylline are described in light of current hypotheses pertaining to leg ulcer development. Also discussed are nine cases in which pentoxifylline, when added to previously unsuccessful local wound care, either improved or healed the refractory ulcers.


Inhibition of growth of Dictyostelium discoideum amoebae by bisphosphonate drugs is dependent on cellular uptake.
The aim of the study was to determine whether bisphosphonates are internalised by Dictyostelium amoebae and whether cellular uptake is required for their growth-inhibitory effects. Bisphosphonates inhibit growth of amoebae of the slime mould Dictyostelium discoideum, by mechanisms that appear to be similar to those that cause inhibition of osteoclastic bone resorption. Cell-free extracts prepared from amoebae that had been incubated with bisphosphonates were analysed by 31P-n.m.r, spectroscopy or ion-exchange f.p.l.c., to identify the presence of bisphosphonates or bisphosphonate metabolites respectively. The growth-inhibitory effect of bisphosphonates towards Dictyostelium amoebae was also examined under conditions in which pinocytosis was inhibited. All of the bisphosphonates studied were internalised by Dictyostelium amoebae, probably by fluid-phase pinocytosis, and could be detected in cell-free extracts. Amoebae that were prevented from internalising bisphosphonates by pinocytosis were markedly resistant to the growth-inhibitory effects of these compounds. In addition, bisphosphonates encapsulated within liposomes were more potent growth inhibitors of Dictyostelium owing to enhanced intracellular delivery of bisphosphonates. All bisphosphonates inhibit Dictyostelium growth by intracellular mechanisms following internalisation of bisphosphonates by fluid-phase pinocytosis. It is therefore likely that bisphosphonates also affect osteoclasts by interacting with intracellular, rather than extracellular, processes.


A fast Monte Carlo algorithm to simulate the spectrum of DNA damages formed by ionizing radiation.
Ionizing radiation produces both singly and multiply damaged DNA sites. Multiply damaged sites (MDS) have been implicated in radiation-induced cell killing and mutagenesis. The spatial distribution of elementary damages (strand breaks and base damages) that constitute MDS is of special interest, since the complexity of MDS has an impact on damage repair. A fast and easy-to-implement algorithm to simulate the local clustering of elementary damages produced by ionizing radiation is proposed. This algorithm captures the major trends in the DNA damage spectrum predicted using detailed track- structure simulations. An attractive feature of the proposed algorithm is that only four adjustable parameters need to be identified to simulate the formation of DNA damage. A convenient recipe to determine the parameters used in the fast Monte Carlo damage simulation algorithm is provided for selected low- and high-LET radiations. The good agreement among the damage yields predicted by the fast and detailed damage formation algorithms suggests that the small-scale spatial distribution of damage sites is determined primarily by independent and purely stochastic events and processes.


Diet quality and survival after ovarian cancer: results from the Women's Health Initiative.
Survival after an ovarian cancer diagnosis is poor. Given the high mortality in these patients, efforts to identify modifiable lifestyle behaviors that could influence survival are needed. Earlier evidence suggests a protective role for vegetables, but no prior studies have evaluated overall dietary quality and ovarian cancer survival. The purpose of this analysis was to evaluate the role of prediagnosis diet quality in ovarian cancer survival. We identified 636 centrally adjudicated cases of ovarian cancer within the Women's Health Initiative Observational Study or Clinical Trials of 161808 postmenopausal women followed from 1995 to 2012. Dietary quality was assessed for the Healthy Eating Index (2005) using a food frequency questionnaire, covariables were obtained from standardized questionnaires, and adiposity was measured by clinic-based measurements of height, weight, and waist circumference. The association between diet quality and mortality was analyzed using Cox proportional hazards regression, adjusted for potential confounders, and stratified by waist circumference, physical activity level, and diabetes status. Tests of statistical significance were two-sided. Overall, higher diet quality was associated with lower all-cause mortality after ovarian cancer (hazard ratio [HR] for highest vs lowest tertile = 0.73; 95% confidence interval [CI] = 0.55 to 0.97, P(trend) = .03). The effect was strongest among women with waist circumference of 88 cm or less and with no history of diabetes (HR = 0.73, 95% CI = 0.54 to 0.98). Physical activity level did not modify the association between diet quality and survival. Our results suggest that overall higher prediagnosis diet quality may protect against mortality after ovarian cancer.