(-)-Epigallocatechin-3-gallate induces apoptosis and inhibits invasion and migration of human cervical cancer cells.
Invasion and metastasis are the major causes of cancer-related death. Pharmacological or therapeutic interventions such as chemoprevention of the progression stages of neoplastic development could result in substantial reduction in the incidence of cancer mortality. (-)-Epigallocatechin-3-gallate (EGCG), a promising chemopreventive agent, has attracted extensive interest for cancer therapy utilizing its antioxidant, anti- proliferative and inhibitory effects on angiogenesis and tumor cell invasion. In this study, we assessed the influence of EGCG on the proliferative potential of HeLa cells by cell viability assay and authenticated the results by nuclear morphological examination, DNA laddering assay and cell cycle analysis. Further we analyzed the anti-invasive properties of EGCG by wound migration assay and gene expression of MMP-9 and TIMP-1 in HeLa cells. Our results indicated that EGCG induced growth inhibition of HeLa cells in a dose- and time- dependent manner. It was observed that cell death mediated by EGCG was through apoptosis. Interestingly, EGCG effectively inhibited invasion and migration of HeLa cells and modulated the expression of related genes (MMP-9 and TIMP-1) . These results indicate that EGCG may effectively suppress promotion and progression stages of cervical cancer development.


Genome-wide methylation in alcohol use disorder subjects: implications for an epigenetic regulation of the cortico-limbic glucocorticoid receptors (NR3C1).
Environmental factors, including substance abuse and stress, cause long-lasting changes in the regulation of gene expression in the brain via epigenetic mechanisms, such as DNA methylation. We examined genome-wide DNA methylation patterns in the prefrontal cortex (PFC, BA10) of 25 pairs of control and individuals with alcohol use disorder (AUD), using the Infinium® MethylationEPIC BeadChip. We identified 5254 differentially methylated CpGs (pnominal < 0.005). Bioinformatic analyses highlighted biological processes containing genes related to stress adaptation, including the glucocorticoid receptor (encoded by NR3C1). Considering that alcohol is a stressor, we focused our attention on differentially methylated regions of the NR3C1 gene and validated the differential methylation of several genes in the NR3C1 network. Chronic alcohol drinking results in a significant increased methylation of the NR3C1 exon variant 1H, with a particular increase in the levels of 5-hydroxymethylcytosine over 5-methylcytosine. These changes in DNA methylation were associated with reduced NR3C1 mRNA and protein expression levels in PFC, as well as other cortico-limbic regions of AUD subjects when compared with controls. Furthermore, we show that the expression of several stress-responsive genes (e.g., CRF, POMC, and FKBP5) is altered in the PFC of AUD subjects. These stress-response genes were also changed in the hippocampus, a region that is highly susceptible to stress. These data suggest that alcohol-dependent aberrant DNA methylation of NR3C1 and consequent changes in other stress-related genes might be fundamental in the pathophysiology of AUD and lay the groundwork for treatments targeting the epigenetic mechanisms regulating NR3C1 in AUD.


Survey of ophthalmic abnormalities in the labradoodle in the UK.
One-hundred and seventy-five labradoodles underwent slit-lamp biomicroscopy and direct and indirect ophthalmoscopy between January 2008 and December 2009. These examinations were performed at shows by the first author. In addition, labradoodle eye certificates from the 2008 and 2009 British Veterinary Association/Kennel Club/International Sheep Dog Society (BVA/KC/ISDS) Eye Scheme were analysed (n=260). The results of the examinations were reviewed, and all ophthalmic abnormalities were recorded. The prevalence of any abnormality was compared with that reported by the Eye Scheme and the KC during 2009 for the labrador retriever, miniature, toy and standard poodles. Multifocal retinal dysplasia (MRD) was identified in 20 labradoodles (4.6 per cent), and cataracts were identified in 16 labradoodles (3.7 per cent). The prevalence of MRD in labradoodles was significantly greater than in labrador retrievers (P<0.0001). There was no difference in the prevalence of cataract between labradoodles and labrador retrievers (P=0.4866). The results of this study suggest that MRD is a relatively common finding in the labradoodle population in the UK. Given such an apparent high prevalence of MRD, routine screening for hereditary eye disease before breeding is advised for this increasingly popular new crossbreed.


Molecular survey of neglected bacterial pathogens reveals an abundant diversity of species and genotypes in ticks collected from animal hosts across Romania.
Ticks are transmitting a wide range of bacterial pathogens that cause substantial morbidity and mortality in domestic animals. The full pathogen burden transmitted by tick vectors is incompletely studied in many geographical areas, and extensive studies are required to fully understand the diversity and distribution of pathogens transmitted by ticks. We sampled 824 ticks of 11 species collected in 19 counties in Romania. Ticks were collected mainly from dogs, but also from other domestic and wild animals, and were subjected to molecular screening for pathogens. Rickettsia spp. was the most commonly detected pathogen, occurring in 10.6% (87/824) of ticks. Several species were detected: Rickettsia helvetica, R. raoultii, R. massiliae, R. monacensis, R. slovaca and R. aeschlimannii. A single occurrence of the zoonotic bacterium Bartonella vinsonii berkhoffii was detected in a tick collected from a dog. Anaplasma phagocytophilum occurred in four samples, and sequences similar to Anaplasma marginale/ovis were abundant in ticks from ruminants. In addition, molecular screening showed that ticks from dogs were carrying an Ehrlichia species identical to the HF strain as well as the enigmatic zoonotic pathogen "Candidatus Neoehrlichia mikurensis". An organism similar to E. chaffeensis or E. muris was detected in an Ixodes ricinus collected from a fox. We describe an abundant diversity of bacterial tick-borne pathogens in ticks collected from animal hosts in Romania, both on the level of species and genotypes/strains within these species. Several findings were novel for Romania, including Bartonella vinsonii subsp. berkhoffii that causes bacteremia and endocarditis in dogs. "Candidatus Neoehrlichia mikurensis" was detected in a tick collected from a dog. Previously, a single case of infection in a dog was diagnosed in Germany. The results warrant further studies on the consequences of tick-borne pathogens in domestic animals in Romania.


Ca(2+)-sensor region of IP(3) receptor controls intracellular Ca(2+) signaling.
Many important cell functions are controlled by Ca(2+) release from intracellular stores via the inositol 1,4,5-trisphosphate receptor (IP(3)R), which requires both IP(3) and Ca(2+) for its activity. Due to the Ca(2+) requirement, the IP(3)R and the cytoplasmic Ca(2+) concentration form a positive feedback loop, which has been assumed to confer regenerativity on the IP(3)-induced Ca(2+) release and to play an important role in the generation of spatiotemporal patterns of Ca(2+) signals such as Ca(2+) waves and oscillations. Here we show that glutamate 2100 of rat type 1 IP(3)R (IP(3)R1) is a key residue for the Ca(2+) requirement. Substitution of this residue by aspartate (E2100D) results in a 10-fold decrease in the Ca(2+) sensitivity without other effects on the properties of the IP(3)R1. Agonist-induced Ca(2+) responses are greatly diminished in cells expressing the E2100D mutant IP(3)R1, particularly the rate of rise of initial Ca(2+) spike is markedly reduced and the subsequent Ca(2+) oscillations are abolished. These results demonstrate that the Ca(2+) sensitivity of the IP(3)R is functionally indispensable for the determination of Ca(2+) signaling patterns.


Molecular imaging with contrast ultrasound and targeted microbubbles.
There is growing interest in the development of methods for imaging cellular and molecular mediators of cardiovascular diseases. Techniques for imaging molecular and cellular alterations have been explored for essentially all noninvasive cardiac imaging modalities. Molecular imaging with contrast-enhanced ultrasound relies on the detection of novel site-targeted microbubble contrast agents. These microbubbles are retained within regions of a specific disease process, thereby allowing phenotypic characterization of tissue. As microbubbles are pure intravascular tracers, the disease processes assessed must be characterized by antigens that are expressed within the vascular compartment. Accordingly, the pathologic states that have been targeted include inflammation, neoplasms, angiogenesis, and thrombus formation, all of which are mediated in part by molecular events within the vascular space. This review describes (1) different strategies that have been used to target microbubbles to regions of disease, (2) the unique challenges for imaging targeted ultrasound contrast agents, and (3) some of the early experience imaging molecular events in animal models of disease.


Regulation of the CTL response by macrophage migration inhibitory factor.
Macrophage migration inhibitory factor (MIF) has been shown to be a pivotal cytokine that mediates host inflammatory and immune responses. Recently, immunoneutralization of MIF has been found to inhibit tumor growth in mice; however, the contributing mechanisms underlying this effect have not been well defined. We investigated whether MIF plays a regulatory role in the expression of CTL activity. In a mouse model of the CTL response using the OVA-transfected tumor cell line EL4 (EG.7), we found that cultures of splenocytes obtained from EG.7-primed mice secrete high levels of MIF following Ag stimulation in vitro. Notably, parallel splenocyte cultures treated with neutralizing anti-MIF mAb showed a significant increase in the CTL response directed against EG.7 cells compared with control mAb-treated cultures. This effect was accompanied by elevated expression of IFN-gamma. Histological examination of the EG. 7 tumors from anti-MIF-treated animals showed a prominent increase in both CD4(+) and CD8(+) T cells as well as apoptotic tumor cells, consistent with the observed augmentation of CTL activity in vivo by anti-MIF. This increased CTL activity was associated with enhanced expression of the common gamma(c)-chain of the IL-2R that mediates CD8(+) T cell survival. Finally, CD8(+) T lymphocytes obtained from the spleens of anti-MIF-treated EG.7 tumor-bearing mice, when transferred into recipient tumor-bearing mice, showed increased accumulation in the tumor tissue. These data provide the first evidence of an important role for MIF in the regulation and trafficking of anti-tumor T lymphocytes in vivo.


Immunoglobulin light-chain amyloidosis: growing recognition, new approaches to therapy, active clinical trials.
Immunoglobulin light-chain amyloidosis needs to be considered in any patient presenting with cardiomyopathy with preserved systolic function, heavy albuminuria, an unexplained sensorimotor peripheral neuropathy, hepatomegaly, or atypical MGUS (monoclonal gammopathy of undetermined significance) or myeloma.The prognosis of the disease is determined by the levels of cardiac biomarkers and the pretreatment levels of immunoglobulin free light chains. All patients with systemic light-chain amyloid require therapy. There is no presymptomatic phase that warrants observation. Stem-cell transplantation produces a high response rate but is a viable option in only 20% of patients. Corticosteroids, alkylating agents, immunomodulatory drugs, and proteasome inhibitors all have shown activity in this disorder, and combinations are currently being explored in clinical trials. Despite advances in the past decade, 30% of patients still die within a year of diagnosis, suggesting that failure to recognize this disorder prior to advanced organ dysfunction remains a major impediment to improving outcomes.


Co-Creating the Cities of the Future.
In recent years, the evolution of urban environments, jointly with the progress of the Information and Communication sector, have enabled the rapid adoption of new solutions that contribute to the growth in popularity of Smart Cities. Currently, the majority of the world population lives in cities encouraging different stakeholders within these innovative ecosystems to seek new solutions guaranteeing the sustainability and efficiency of such complex environments. In this work, it is discussed how the experimentation with IoT technologies and other data sources form the cities can be utilized to co-create in the OrganiCity project, where key actors like citizens, researchers and other stakeholders shape smart city services and applications in a collaborative fashion. Furthermore, a novel architecture is proposed that enables this organic growth of the future cities, facilitating the experimentation that tailors the adoption of new technologies and services for a better quality of life, as well as agile and dynamic mechanisms for managing cities. In this work, the different components and enablers of the OrganiCity platform are presented and discussed in detail and include, among others, a portal to manage the experiment life cycle, an Urban Data Observatory to explore data assets, and an annotations component to indicate quality of data, with a particular focus on the city-scale opportunistic data collection service operating as an alternative to traditional communications.


Induction of host defences by Rhizobium during ineffective nodulation of pea (Pisum sativum L.) carrying symbiotically defective mutations sym40 (PsEFD), sym33 (PsIPD3/PsCYCLOPS) and sym42.
Rhizobia are able to establish a beneficial interaction with legumes by forming a new organ, called the symbiotic root nodule, which is a unique ecological niche for rhizobial nitrogen fixation. Rhizobial infection has many similarities with pathogenic infection and induction of defence responses accompanies both interactions, but defence responses are induced to a lesser extent during rhizobial infection. However, strong defence responses may result from incompatible interactions between legumes and rhizobia due to a mutation in either macro- or microsymbiont. The aim of this research was to analyse different plant defence reactions in response to Rhizobium infection for several pea (Pisum sativum) mutants that result in ineffective symbiosis. Pea mutants were examined by histochemical and immunocytochemical analyses, light, fluorescence and transmission electron microscopy and quantitative real-time PCR gene expression analysis. It was observed that mutations in pea symbiotic genes sym33 (PsIPD3/PsCYCLOPS encoding a transcriptional factor) and sym40 (PsEFD encoding a putative negative regulator of the cytokinin response) led to suberin depositions in ineffective nodules, and in the sym42 there were callose depositions in infection thread (IT) and host cell walls. The increase in deposition of unesterified pectin in IT walls was observed for mutants in the sym33 and sym42; for mutant in the sym42, unesterified pectin was also found around degrading bacteroids. In mutants in the genes sym33 and sym40, an increase in the expression level of a gene encoding peroxidase was observed. In the genes sym40 and sym42, an increase in the expression levels of genes encoding a marker of hypersensitive reaction and PR10 protein was demonstrated. Thus, a range of plant defence responses like suberisation, callose and unesterified pectin deposition as well as activation of defence genes can be triggered by different pea single mutations that cause perception of an otherwise beneficial strain of Rhizobium as a pathogen.


Ethics and policy: dealing with public attitudes.
The current trend towards ethical scrutiny and oversight is very much a social trend. Many of the results of this trend are perfectly reasonable but some go harmfully too far. In this paper, caution is advocated about public attitudes and social trends. Although there is often a degree of truth in them, there is an inevitable simplification of the issues involved. The more specific danger for the professions is to think that public attitudes and social trends simply deliver 'the ethical'. In this context a more adequate account of ethics is considered--one that is relevant for professions like radiology confronting the demands of ethical scrutiny and oversight. The paper concludes with some suggestions about how to incorporate the important aspects of public attitudes and social trends without being subservient to them.


Predictors of motor complications in early Parkinson's disease: A prospective cohort study.
The objective of this study was to identify clinical predictors of motor complications (dyskinesia and motor fluctuations) of levodopa in a prospectively recruited PD cohort using longitudinal analysis. An inception cohort (Oxford Discovery) of 734 patients was followed to a maximum of 10 years from diagnosis using a discrete-time survival analysis. A subset analysis was used to validate an online dyskinesia-risk calculator developed from the results of the Stalevo Reduction in Dyskinesia Evaluation PD trial. A total of 186 cases of dyskinesia and 254 cases of motor fluctuations were observed. Dyskinesia incidence increased with time (risk per 100 participants [95% confidence interval] 13 [11-16] <3.5 years, 16 [13-21] 3.5-5.0 years, 19 [14-26] 5-6.5 years, and 23 [16-33] >6.5 years from diagnosis). Motor complication predictors were grouped as medication predictors, disease predictors and patient predictors. Baseline nonmotor feature severity, low mood, anxiety, and age at symptom onset were associated with motor complications among a number of previously identified predictors. Replication of the Stalevo Reduction in Dyskinesia Evaluation PD calculator was reasonable with the area under the curve for dyskinesia risk score as a predictor of dyskinesia being 0.68 (95% confidence interval, 0.55-0.81). This study quantifies risk of motor complications, finds consistent predictors, and demonstrates the novel finding that nonmotor features of PD, particularly low mood and anxiety, are significant risk factors for motor complications. Further validation of dyskinesia risk scores are required as well as evidence to determine if the routine use of such scores can be clinically valuable in enhancing patient care and quality of life. © 2019 The Authors. Movement Disorders published by Wiley Periodicals, Inc. on behalf of International Parkinson and Movement Disorder Society.


Long-term Outcomes of Critically Ill Patients With Stroke Requiring Mechanical Ventilation.
Data on outcomes of critically ill patients requiring mechanical ventilation at the onset of stroke are limited. To assess the hospital and long-term functional outcomes of patients with stroke who require mechanical ventilation. This retrospective single-center cohort study performed from 1994 to 2008 involved adult patients within 7 days of stroke onset and who required intensive care unit admission and mechanical ventilation. A total of 274 patients requiring mechanical ventilation at the onset of stroke were analyzed. Indications for intubation included coma in 195 patients (71%). The median (interquartile range) score on the Glasgow Coma Scale at admission to the intensive care unit was 6 (3-9). Forty-four patients (16%) had sepsis at intensive care unit admission. The overall hospital mortality rate was 53%. After adjustment for confounders, severity of illness at admission as assessed by the Simplified Acute Physiology Score II (odds ratio, 1.07; 95% CI, 1.05-1.10), anisocoria (odds ratio, 5.26; 95% CI, 1.76-15.80), and sepsis at intensive care unit admission (odds ratio, 0.40; 95% CI, 0.19-0.85) were associated with outcome. At 1 year, median (interquartile range) modified Rankin Scale score was 6 (2-6). Only 89 patients (32%) exhibited mild to moderate neurologic impairment. In this study, adult patients requiring mechanical ventilation at the onset of stroke experienced high 1-year mortality, with survivors having poor functional status.


Osteogenic response to BMP-2 of hMSCs grown on apatite-coated scaffolds.
Osteoconductive materials play a critical role in promoting integration with surrounding bone tissue and resultant bone repair in vivo. However, the impact of 3D osteoconductive substrates coupled with soluble signals on progenitor cell differentiation is not clear. In this study, we investigated the influence of bone morphogenetic protein-2 (BMP-2) concentration on the osteogenic differentiation of human mesenchymal stem cells (hMSCs) when seeded in carbonated apatite-coated polymer scaffolds. Mineralized scaffolds were more hydrophilic and adsorbed more BMP-2 compared to non-mineralized scaffolds. Changes in alkaline phosphatase (ALP) activity within stimulated hMSCs were dependent on the dose of BMP-2 and the scaffold composition. We detected more cell-secreted calcium on mineralized scaffolds at all time points, and higher BMP-2 concentrations resulted in increased ALP and calcium levels. RUNX2 and IBSP gene expression within hMSCs was affected by both substrate and soluble signals, SP7 by soluble factors, and SPARC by substrate-mediated cues. The present data indicate that a combination of apatite and BMP-2 do not simply enhance the osteogenic response of hMSCs, but act through multiple pathways that may be both substrate- and growth factor-mediated. Thus, multiple signaling strategies will likely be necessary to achieve optimal bone regeneration.


Low dose of propranolol down-modulates bone resorption by inhibiting inflammation and osteoclast differentiation.
Bones are widely innervated, suggesting an important role for the sympathetic regulation of bone metabolism, although there are controversial studies. We investigated the effects of propranolol in a model of experimental periodontal disease. Rats were assigned as follows: animals without ligature; ligated animals receiving vehicle and ligated animals receiving 0.1, 5 or 20 mg·kg(-1) propranolol. After 30 days, haemodynamic parameters were measured by cardiac catheterization. Gingival tissues were removed and assessed for IL-1β, TNF-α and cross-linked carboxyterminal telopeptides of type I collagen (CTX) by elisa, or intercellular adhesion molecule 1 (ICAM-1), receptor activator of NF-κ B ligand (RANKL) and osteoprotegerin (OPG) by Western blot analysis. Sections from the mandibles were evaluated for bone resorption. Also, we analysed the ability of propranolol to inhibit osteoclastogenesis in vitro. Propranolol at 0.1 and 5 mg·kg(-1) reduced the bone resorption as well as ICAM-1 and RANKL expression. However, only 0.1 mg·kg(-1) reduced IL-1β, TNF-α and CTX levels as well as increased the expression of OPG, but did not alter any of the haemodynamic parameters. Propranolol also suppressed in vitro osteoclast differentiation and resorptive activity by inhibiting the nuclear factor of activated T cells (NFATc)1 pathway and the expression of tartrate-resistant acid phosphatase (TRAP), cathepsin K and MMP-9. Low doses of propranolol suppress bone resorption by inhibiting RANKL-mediated osteoclastogenesis as well as inflammatory markers without affecting haemodynamic parameters.


Evidence for curricular and instructional design approaches in undergraduate medical education: An umbrella review.
An umbrella review compiles evidence from multiple reviews into a single accessible document. This umbrella review synthesizes evidence from systematic reviews on curricular and instructional design approaches in undergraduate medical education, focusing on learning outcomes. We conducted bibliographic database searches in Medline, EMBASE and ERIC from database inception to May 2013 inclusive, and digital keyword searches of leading medical education journals. We identified 18,470 abstracts; 467 underwent duplicate full-text scrutiny. Thirty-six articles met all eligibility criteria. Articles were abstracted independently by three authors, using a modified Kirkpatrick model for evaluating learning outcomes. Evidence for the effectiveness of diverse educational approaches is reported. This review maps out empirical knowledge on the efficacy of a broad range of educational approaches in medical education. Critical knowledge gaps, and lapses in methodological rigour, are discussed, providing valuable insight for future research. The findings call attention to the need for adopting evaluative strategies that explore how contextual variabilities and individual (teacher/learner) differences influence efficacy of educational interventions. Additionally, the results underscore that extant empirical evidence does not always provide unequivocal answers about what approaches are most effective. Educators should incorporate best available empirical knowledge with experiential and contextual knowledge.


Pre-saccadic perception: Separate time courses for enhancement and spatial pooling at the saccade target.
We interact with complex scenes using eye movements to select targets of interest. Studies have shown that the future target of a saccadic eye movement is processed differently by the visual system. A number of effects have been reported, including a benefit for perceptual performance at the target ("enhancement"), reduced influences of backward masking ("un-masking"), reduced crowding ("un-crowding") and spatial compression towards the saccade target. We investigated the time course of these effects by measuring orientation discrimination for targets that were spatially crowded or temporally masked. In four experiments, we varied the target-flanker distance, the presence of forward/backward masks, the orientation of the flankers and whether participants made a saccade. Masking and randomizing flanker orientation reduced performance in both fixation and saccade trials. We found a small improvement in performance on saccade trials, compared to fixation trials, with a time course that was consistent with a general enhancement at the saccade target. In addition, a decrement in performance (reporting the average flanker orientation, rather than the target) was found in the time bins nearest saccade onset when random oriented flankers were used, consistent with spatial pooling around the saccade target. We did not find strong evidence for un-crowding. Overall, our pattern of results was consistent with both an early, general enhancement at the saccade target and a later, peri-saccadic compression/pooling towards the saccade target.


1306-km 20x124.8-Gb/s PM-64QAM transmission over PSCF with net SEDP 11,300 (b ∙ km)/s/Hz using 1.15 samp/symb DAC.
We demonstrated the transmission of a Nyquist-WDM signal based on PM-64QAM modulation in an EDFA-only submarine configuration composed of 54.4 km-long fiber spans: 20 channels at 124.8-Gb/s were propagated over 1306 km of low-loss pure-silica-core fiber (PSCF). Thanks to an aggressive digital spectral shaping, we achieved a raw spectral efficiency (SE) of 10.4 b/s/Hz, corresponding to 8.67 b/s/Hz net SE when considering a 20% FEC overhead. Transmitter DACs are operated at a record-low 1.15 samples/symbol, enabled by the insertion of advanced anti-alias filters. The achieved SE-times-distance product was 11,327 (b ∙ km)/(s ∙ Hz), the highest reported so far for PM-64QAM. Combining the experimental results with the performance predictions obtained using an analytical model of nonlinear propagation in uncompensated coherent optical systems (the so-called "GN-model"), we show that PM-64QAM is a realistic option for ultra-high capacity systems in the 1,000 km range, carrying up 40 Tb/s in the C-band.


Reconstruction and minimal gene requirements for the alternative iron-only nitrogenase in Escherichia coli.
All diazotrophic organisms sequenced to date encode a molybdenum-dependent nitrogenase, but some also have alternative nitrogenases that are dependent on either vanadium (VFe) or iron only (FeFe) for activity. In Azotobacter vinelandii, expression of the three different types of nitrogenase is regulated in response to metal availability. The majority of genes required for nitrogen fixation in this organism are encoded in the nitrogen fixation (nif) gene clusters, whereas genes specific for vanadium- or iron-dependent diazotophy are encoded by the vanadium nitrogen fixation (vnf) and alternative nitrogen fixation (anf) genes, respectively. Due to the complexities of metal-dependent regulation and gene redundancy in A. vinelandii, it has been difficult to determine the precise genetic requirements for alternative nitrogen fixation. In this study, we have used Escherichia coli as a chassis to build an artificial iron-only (Anf) nitrogenase system composed of defined anf and nif genes. Using this system, we demonstrate that the pathway for biosynthesis of the iron-only cofactor (FeFe-co) is likely to be simpler than the pathway for biosynthesis of the molybdenum-dependent cofactor (FeMo-co) equivalent. A number of genes considered to be essential for nitrogen fixation by FeFe nitrogenase, including nifM, vnfEN, and anfOR, are not required for the artificial Anf system in E. coli. This finding has enabled us to engineer a minimal FeFe nitrogenase system comprising the structural anfHDGK genes and the nifBUSV genes required for metallocluster biosynthesis, with nifF and nifJ providing electron transport to the alternative nitrogenase. This minimal Anf system has potential implications for engineering diazotrophy in eukaryotes, particularly in compartments (e.g., organelles) where molybdenum may be limiting.


Arcs and Clumps in the Uranian lgr Ring.
Careful reprocessing of the Voyager images reveals that the Uranìan lambda ring has marked longitudinal variations in brightness comparable in magnitude to those in Saturn's F ring and Neptune's Adams ring. The ring's variations show a dominant five-cycle (72-degree) periodicity, although additional structure down to scales of about 0.5 degree is also present. The ring's shape is defined by a small overall eccentricity plus a six-cycle (60-degree) sinusoidal variation of radial amplitude around 4 kilometers. Both of these properties can be explained by the resonant perturbations of a moon at a semimajor axis of 56,479 kilometers, but no known moon orbits at this location. Unfortunately, the mass required suggests that such a body should have been imaged by Voyager.


Mortality among participants in the Multicenter AIDS Cohort Study and the Women's Interagency HIV Study.
Many studies that have reported decreases in human immunodeficiency virus (HIV)-related mortality since the advent of highly active antiretroviral therapy (HAART) have also reported steady increases in non-HIV-related mortality over the same time periods. We examined temporal trends and risk factors for accident- and injury-related mortality among HIV-infected and -uninfected participants in the Women's Interagency HIV Study (WIHS) and the Multicenter AIDS Cohort Study (MACS). Information on causes of death was recorded for all participants in the MACS and WIHS cohort studies who died, and causes of death were categorized as accident- or injury-related deaths or not. Mortality rates were calculated by time periods, prior to the widespread use of HAART (before 1997) and after (1997-2002), and risk factors. Cause of death information was available for 619 women in the WIHS who died (during the period 1994-2002) and 1830 men in the MACS who died (during the period 1984-2002). The death rates were higher for accident- or injury-related mortality in the WIHS (2.96 deaths per 1000 person-years for the HIV-infected group and 2.96 per 1000 person-years for the HIV-uninfected group), compared with the participants in MACS (0.79 deaths per 1000 person-years for the HIV-infected group and 0.63 per 1000 person-years for the HIV-uninfected group). In the final multivariate analysis, the following factors were associated with significantly higher risk in men: higher education, depressive symptoms, and a greater number of sex partners. Among women, the significant risk factors for death were decreased CD4+ T cell count, unemployment, higher alcohol use, and injection drug use. The characteristics of the men in the MACS who died and women in the WIHS who died differ, as do the risk factors for mortality. These results characterize important target groups for interventions to reduce accident- and injury-related deaths.


CNS location of uterine-related neurons revealed by trans-synaptic tracing with pseudorabies virus and their relation to estrogen receptor-immunoreactive neurons.
Retrograde, transneuronal tracing with Bartha's strain of pseudorabies virus was used in rats to identify spinal cord, brainstem and hypothalamic loci of uterine-related neurons that could function in the regulation of uterine activity. Based on the premise that estrogen might influence such uterine-related neurons, the existence of estrogen receptors in neurons in these same loci was examined. Viral injections were made into the uterine cervix, body and cervical end of the uterine horns, and the rats allowed to survive for four to six days. After four days, mainly the spinal cord, medulla and pons contained virus-infected neurons. After longer survival times, progressively higher levels of the neuraxis contained viral-labeled neurons, so that by six days hypothalamic uterine-related neurons were identified. First-order virus-infected neurons were visualized by immunohistochemistry in the pelvic paracervical parasympathetic ganglia and in inferior mesenteric sympathetic ganglia. Preganglionic and putative interneurons were labeled in the lumbosacral spinal cord and thoracic spinal cord mainly in the lateral horn area (sacral parasympathetic nucleus and intermediolateral nucleus), lateral aspect of the dorsal horn, intermediate gray, lamina X and dorsal gray commissural area. In the brainstem, labeling was most evident and consistent in the nucleus tractus solitarius, ventrolateral medulla, raphe magnus and pallidus nuclei, parapyramidal area, A5 cell group, Barrington's nucleus of the pons and periaqueductal gray of the midbrain. In the hypothalamus, virus-infected neurons were most marked in the paraventricular nucleus, with fewer in the medial preoptic area and ventromedial hypothalamic nucleus. Estrogen receptor-immunoreactive neurons were most often present among the virus-labeled uterine-related neurons of the spinal cord, nucleus tractus solitarius, ventrolateral medulla, periaqueductal gray, medial preoptic area and ventromedial hypothalamic nucleus. These results identify a multisynaptic pathway of neurons whose eventual output is involved in uterine functions, whose distribution is similar to that revealed by pseudorabies virus tracing from other visceral organs, and which are often mixed among estrogen-responsive neurons.


Quantifying reproducibility in computational biology: the case of the tuberculosis drugome.
How easy is it to reproduce the results found in a typical computational biology paper? Either through experience or intuition the reader will already know that the answer is with difficulty or not at all. In this paper we attempt to quantify this difficulty by reproducing a previously published paper for different classes of users (ranging from users with little expertise to domain experts) and suggest ways in which the situation might be improved. Quantification is achieved by estimating the time required to reproduce each of the steps in the method described in the original paper and make them part of an explicit workflow that reproduces the original results. Reproducing the method took several months of effort, and required using new versions and new software that posed challenges to reconstructing and validating the results. The quantification leads to "reproducibility maps" that reveal that novice researchers would only be able to reproduce a few of the steps in the method, and that only expert researchers with advance knowledge of the domain would be able to reproduce the method in its entirety. The workflow itself is published as an online resource together with supporting software and data. The paper concludes with a brief discussion of the complexities of requiring reproducibility in terms of cost versus benefit, and a desiderata with our observations and guidelines for improving reproducibility. This has implications not only in reproducing the work of others from published papers, but reproducing work from one's own laboratory.


Development and Validation of the Stalking and Obsessive Relational Intrusions Questionnaire (SORI-Q).
Stalking and obsessive relational intrusions both refer to a pervasive and unwanted pattern of pursuit behaviors, the former being a criminal offense evoking fear and a sense of menace in the victim, while the latter may be perceived as annoying or otherwise undesirable, but not necessarily fear inducing. While the individual and societal costs of stalking and obsessive relational intrusion are increasingly recognized, research regarding these behaviors and their consequences has been limited by measurement issues, as most studies have relied on questionnaires and checklists based on very limited validation data. The goal of the present study is to report on the development and validation of the Stalking and Obsessive Relational Intrusions Questionnaire (SORI-Q), a 28-item self-report questionnaire designed to probe for perpetration of stalking-like behaviors. Young adults (age 18-30 years) from a community sample (N = 1,804; 82.6% women) were recruited online. They completed the SORI-Q, along with measures of dark personality traits, insecure attachment dimensions, and intimate partner violence. Overall, the SORI-Q displayed sound psychometric properties. Exploratory and Confirmatory Factor Analysis yielded a two-factor solution (Hyper-intimacy and Domineering control) with adequate to good fit indices. The total scale and the two factor scores showed high internal consistency (above 0.70 for all indices). A number of gender differences were observed at total-, factor-, and item-level, the most outstanding being that women had a higher score on the total SORI-Q score, and on the Domineering control factor and most of its items. The questionnaire showed conceptually meaningful positive correlations with dark personality traits, attachment anxiety, and intimate partner violence. Dominance analysis revealed that attachment anxiety and Machiavellianism were the strongest statistical predictors of SORI-Q scores. The SORI-Q should be seen as a promising new measure of stalking-like and ORI behaviors in young adults from community settings.


Uncovering Natural Longevity Alleles from Intercrossed Pools of Aging Fission Yeast Cells.
Quantitative traits often show large variation caused by multiple genetic factors . One such trait is the chronological lifespan of non-dividing yeast cells, serving as a model for cellular aging. Screens for genetic factors involved in aging typically assay mutants of protein-coding genes. To identify natural genetic variants contributing to cellular aging, we exploited two strains of the fission yeast, Schizosaccharomyces pombe, that differ in chronological lifespan. We generated segregant pools from these strains and subjected them to advanced intercrossing over multiple generations to break up linkage groups. We chronologically aged the intercrossed segregant pool, followed by genome sequencing at different times to detect genetic variants that became reproducibly enriched as a function of age. A region on Chromosome II showed strong positive selection during aging. Based on expected functions, two candidate variants from this region in the long-lived strain were most promising to be causal: small insertions and deletions in the 5'-untranslated regions of ppk31 and SPBC409.08 Ppk31 is an ortholog of Rim15, a conserved kinase controlling cell proliferation in response to nutrients, while SPBC409.08 is a predicted spermine transmembrane transporter. Both Rim15 and the spermine-precursor, spermidine, are implicated in aging as they are involved in autophagy-dependent lifespan extension. Single and double allele replacement suggests that both variants, alone or combined, have subtle effects on cellular longevity. Furthermore, deletion mutants of both ppk31 and SPBC409.08 rescued growth defects caused by spermidine. We propose that Ppk31 and SPBC409.08 may function together to modulate lifespan, thus linking Rim15/Ppk31 with spermidine metabolism.


A cluster randomized controlled trial comparing the effectiveness of an individual planning intervention with collaborative planning in adolescent friendship dyads to enhance physical activity (TWOgether).
Most adolescents do not meet the recommendations for physical activity (PA) of at least 1 h per day. Individual planning (IP) interventions, including forming plans for when, where and how (action planning) to engage in a behavior, as well as the planning for how to deal with arising barriers (coping planning), are effective to enhance PA in adults. Collaborative planning (CP) is conjoint planning of two individuals regarding a behavior which is performed together. It is assumed that CP stimulates social exchange processes between the planning partners. However, it remains unclear whether planning interventions of PA in adolescents are successful and which planning intervention is more effective. Thus, this cluster randomized controlled trial (RCT) examines changes in daily moderate-to-vigorous PA in adolescents' friendship dyads resulting from planning. Individual self-regulating mechanism and social exchange processes are proposed as mediating mechanisms of the effects of planning for health behavior change. A single-blind four-arm parallel-group cluster-RCT is used. The sample consists of 400 friendship dyads between 14 and 18 years of age. As the recruitment takes place in schools, a cluster randomization of the schools is used to enroll dyads to (a) an IP intervention, (b) a CP intervention or (c) one of the two no-planning control conditions. Devise-measured and self-reported PA as the primary outcomes, self-regulatory strategies, and social exchange processes as secondary outcomes are assessed at three or four time points. After baseline measurement, the baseline ecological momentary assessment of the main variables takes place for 8 days followed by the intervention and a 7-days diary phase. Follow-ups are 1 month and 6 months later. Subsequent to the six-month follow-up, another 7-days diary phase takes place. This is the first study examining IP in comparison to CP in adolescents applying a single-blind cluster RCT. Consequently, the study allows for understanding the efficacy of individual and collaborative planning and the underlying mechanisms in adolescent dyads. This RCT was funded by the Swiss National Science Foundation (100019_169781/1) and was registered on 18/06/2018 at ClinicalTrials.gov : NCT03575559 .


Disseminated Penicillium marneffei infection presenting as a right upper lobe mass in an HIV positive patient.
A 35 year old HIV positive patient from Hong Kong presented with a fever, cough and a skin rash in association with a lung mass, all of which were due to disseminated Penicillium marneffei infection. He made a good response to antifungal therapy. The lung mass is a previously undescribed pulmonary manifestation of disseminated Penicillium marneffei infection. Infections with this fungus should be suspected in any patient with HIV and respiratory symptoms who has visited southeast Asia.


Blunted sensitivity to sucrose in autoimmune MRL-lpr mice: a curve-shift study.
Lupus-prone MRL-lpr mice show an autoimmunity-associated behavioral syndrome that has many features similar to the effects of chronic stress. The present study evaluated whether autoimmune MRL-lpr mice show reduced responsiveness to sucrose, as observed in normal animals exposed to chronic mild stress. Sixteen-week old MRL-lpr mice and their age-matched congenic MRL +/+ controls were given 0%, 0.5%, 1%, 2%, 4%, 8%, or 16% sucrose solution to drink every 48 h in a one-bottle test. The MRL-lpr mice drank less than controls at all concentrations, except at 16%. The amount of sucrose consumed vs. solution concentration followed a saturation curve. Estimates were obtained for the concentration yielding the half-maximum response (X50) and the response at saturating concentration of sucrose (Rmax). The X50 was significantly higher in MRL-lpr than in MRL +/+ mice, indicating a shift to the right of the concentration-intake curve. The Rmax did not differ significantly between substrains, suggesting that the autoimmune process did not affect performance capacity. Pretreatment with the immunosuppressant cyclophosphamide diminished the substrain difference in X50, suggesting that reduced sensitivity to sucrose is related to autoimmune/inflammatory factors. These results support the similarity between autoimmunity-associated behavioral syndrome and behavioral changes produced by chronic stress, and suggest common neuroendocrine mechanisms. Because reduced sensitivity to palatable stimulus may reflect blunted hedonic responsiveness ("anhedonia"), it is hypothesized that an autoimmune/inflammatory factor(s) produces the depression found in human lupus, and some cases of affective disorder.


Bile acids induce glucagon-like peptide 2 secretion with limited effects on intestinal adaptation in early weaned pigs.
Early weaning is a stressful event characterized by a transient period of intestinal atrophy that may be mediated by reduced secretion of glucagon-like peptide (GLP) 2. We tested whether enterally fed bile acids or plant sterols could increase nutrient-dependent GLP-2 secretion and improve intestinal adaptation in weanling pigs. During the first 6 d after weaning, piglets were intragastrically infused once daily with either deionized water (control), chenodeoxycholic acid (CDC; 60 mg/kg body weight), or β-sitoesterol (BSE; 100 mg/kg body weight). Infusing CDC increased plasma GLP-2 (P < 0.05) but did not affect plasma GLP-1 and feed intake. The intestinal expression of glucagon-like peptide 2 receptor, sodium-dependent bile acid transporter, farnesoid X receptor, and guanosine protein-coupled bile acid receptor genes were not affected by CDC treatment. The intragastric administration of CDC did not alter the weight and length of the intestine, yet increased the activation of caspase-3 in ileal villi (P < 0.02) and the expression of interleukin 6 (P < 0.002) in the jejunum. In contrast, infusing BSE did not affect any of the variables that were measured. Our results show that the enteral administration of the bile acid CDC potentiates the nutrient-induced secretion of endogenous GLP-2 in early-weaned pigs. Bile acid-enhanced release of GLP-2, however, did not result in improved intestinal growth, morphology, or inflammation during the postweaning degenerative phase.


[Headache in cervical syndrome].
Headache is a common symptom in patients suffering from cervical spine disorders. The percentage of headaches in association with degenerative changes of the cervical spine ranges from 13 to 79% and that in association with indirect trauma of the cervical spine from 48 to 82%. Based on neuroanatomical and neurophysiological studies, the relationship of the upper cervical spine and the trigeminal nuclei has been demonstrated and serves as an explanation for perceived head pain in cervical spine disorders. As a source of pain, tension in the suboccipital muscles, irritation of the third occipital nerve, and degenerative changes of the C2/C3 joints have been discussed. Bogduk, in his studies, asserts a direct causative role of mechanical derangement of the cervical spine in the pathogenesis of cervicogenic headaches. In 1983, Sjaastad et al. postulated the concept of 'cervicogenic headaches': a migraine-like headache due to certain disorders of the cervical spine, strictly located unilaterally, its manifestations being in the temporal, frontal, and ocular areas, with associated symptoms such as slight lacrimation, conjunctival injections, tinnitus, runny nose, and erythema in the forehead ipsilaterally. As arguments in favour of a cervical origin, Sjaastad mentioned the following features: precipitation of the headaches either by neck movements or by pressure against certain tender spots on the neck, the possibility of homolateral shoulder or arm pain, stiffness and pain of the neck, and reduced mobility of the cervical spine. In 1988, the Headache Classification Committee of the International Headache Society set strict criteria for 'headaches' to be classified as to be of cervical origin.


Genotype-phenotype relationships in U.S. melanoma-prone families with CDKN2A and CDK4 mutations.
Two genes have been implicated in the development of cutaneous malignant melanoma (CMM). CDK4 (the gene encoding cyclin-dependent kinase 4, an oncogene) has exhibited germline mutations found in only three melanoma-prone families to date. CDKN2A is a tumor suppressor gene that encodes p16 (which inhibits activity of the cyclin D1-CDK4 complex) with germline mutations detected in 10%-25% of melanoma-prone families, some of whom are also prone to pancreatic cancer. We compared 104 CMM patients from 17 CDKN2A families and 12 CMM case subjects from two CDK4 families. We used nonparametric statistics to test for differences in median age at first CMM diagnosis, numbers of CMMs, and numbers of nevi. The three recurrent mutations were haplotyped. All P values were two-sided. The median age at CMM diagnosis (P =.70) and the median numbers of CMMs (P =.73) did not differ between CMM case subjects from CDKN2A versus CDK4 families. Assessment of CMM case subjects from CDKN2A families with and without pancreatic cancer revealed no statistically significant differences in median age at diagnosis (P =.80) or in tumor number (P =.24). There was, however, a statistically significant difference in age-adjusted median numbers of nevi (P =.004), and CMM case subjects from CDKN2A families without pancreatic cancer had greater numbers of nevi. Recurrent CDKN2A mutations were a change from valine to aspartic acid at codon 126 (n = 3) and from glycine to tryptophan at codon 101 (n = 3). Six CDKN2A families had pancreatic cancer. Both CDK4 families carried a mutation resulting in an arginine-to-cysteine substitution at codon 24. Analyses of recurrent CDKN2A and CDK4 mutations suggested common haplotypes. The recurrent CDKN2A mutations were observed in families with and without pancreatic cancer, which suggests that other factors may be involved in the development of pancreatic cancer. Despite hypothetical differences in the mechanisms of action between CDKN2A and CDK4, clinical factors were indistinguishable between CMM case subjects from CDKN2A versus CDK4 families.


North American SOLITAIRE Stent-Retriever Acute Stroke Registry: choice of anesthesia and outcomes.
Previous work that predated the availability of the safer stent-retriever devices has suggested that general anesthesia (GA) may have a negative impact on outcomes in patients with acute ischemic stroke undergoing endovascular therapy. We reviewed demographic, clinical, procedural (GA versus local anesthesia [LA], etc), and site-adjudicated angiographic and clinical outcomes data from consecutive patients treated with the Solitaire FR device in the investigator-initiated North American SOLITAIRE Stent-Retriever Acute Stroke (NASA) Registry. The primary outcomes were 90-day modified Rankin Scale, mortality, and symptomatic intracranial hemorrhage. A total of 281 patients from 18 centers were enrolled. GA was used in 69.8% (196/281) of patients. Baseline demographic and procedural factors were comparable between the LA and GA groups, except the former demonstrated longer time-to-groin puncture (395.4±254 versus 337.4±208 min; P=0.04), lower National Institutes of Health Stroke Scale (NIHSS; 16.2±5.8 versus 18.8±6.9; P=0.002), lower balloon-guide catheter usage (22.4% versus 49.2%; P=0.0001), and longer fluoroscopy times (39.5±33 versus 28±22.8 min; P=0.008). Recanalization (thrombolysis in cerebral infarction ≥2b; 72.94% versus 73.6%; P=0.9) and rate of symptomatic intracranial hemorrhage (7.1% versus 11.2%; P=0.4) were similar but modified Rankin Scale ≤2 was achieved in more LA patients, 52.6% versus 35.6% (odds ratio, 1.4 [1.1-1.8]; P=0.01). In multivariate analysis, hypertension, NIHSS, unsuccessful revascularization, and GA use (odds ratio, 3.3 [1.6-7.1]; P=0.001) were associated with death. When only anterior circulation and elective GA patients were included, there was a persistent difference in good outcomes in favor of LA patients (50.7% versus 35.5%; odds ratio, 1.3 [1.01-1.6]; P=0.04). The NASA Registry has demonstrated that clinical outcomes and survival are significantly better in patients treated with LA, without increased symptomatic intracranial hemorrhage risk. Future trials should prospectively evaluate the effect of GA on outcomes.


Development of a ReaxFF reactive force field for lithium ion conducting solid electrolyte Li1+xAlxTi2-x(PO4)3 (LATP).
We developed a ReaxFF reactive force field for NASICON-type Li1+xAlxTi2-x(PO4)3 (LATP) materials, which is a promising solid-electrolyte that may enable all-solid-state lithium-ion batteries. The force field parameters were optimized based on density functional theory (DFT) data, including equations of state and the heats of formation of ternary metal oxides and metal phosphate crystal phases (e.g., LixTiO2, Al2TiO5, LiAlO2, AlPO4, Li3PO4 and LiTi2(PO4)3 (LTP)), and the energy barriers for Li diffusion in TiO2 and LTP via vacancies and interstitial sites. Using ReaxFF, the structural and the energetic features of LATP were described properly across various compositions - Li occupies more preferentially the interstitial site next to Al than next to Ti. Also, as observed in experimental data, the lattice parameters decrease when Ti is partly substituted by Al because of the smaller size of the Al cation. Using this force field, the diffusion mechanism and the ionic conductivity of Li in LTP and LATP were investigated at T = 300-1100 K. Low ionic conductivity (5.9 × 10-5 S cm-1 at 300 K) was obtained in LTP as previously reported. In LATP at x = 0.2, the ionic conductivity was slightly improved (8.4 × 10-5 S cm-1), but it is still below the experimental value, which is on the order of 10-4 to 10-3 S cm-1 at x = 0.3-0.5. At higher x (higher Al composition), LATP has a configurational diversity due to the Al substitution and the concomitant insertion of Li. By performing a hybrid MC/MD simulation for LATP at x = 0.5, a thermodynamically stable LATP configuration was obtained. The ionic conductivity of this LATP configuration was calculated to be 7.4 × 10-4 S cm-1 at 300 K, which is one order of magnitude higher than the ionic conductivity for LTP and LATP at x = 0.2. This value is in good agreement with our experimental value (2.5 × 10-4 S cm-1 at 300 K) and the literature values. The composition-dependent ionic conductivity of LATP was successfully demonstrated using the ReaxFF reactive force field, verifying the applicability of the LATP force field for the understanding of Li diffusion and the design of highly Li ion conductive solid electrolytes. Furthermore, our results also demonstrate the feasibility of the MC/MD method in modeling LATP configuration, and provide compelling evidence for the solid solution sensitivity on ionic conductivity.


Harnessing endogenous signals from hepatocytes using a low volume multi-well plate.
Hepatocytes are highly differentiated epithelial cells that lose their phenotype and function when removed from the in vivo environment. Given the importance of hepatic cultures for drug toxicity, bioartificial liver assist devices and basic biology studies, considerable efforts have been focused on the maintenance of hepatic function in vitro. The methods used to date include co-cultivation of hepatocytes with stromal cells, organizing these cells into spheroids and imbedding them into bioactive gels. Our team has recently demonstrated that primary rat hepatocytes confined to microfluidic channels in the absence of convection maintained the epithelial phenotype through upregulation of endogenous signals including hepatocyte growth factor (HGF). The objective of the present study was to transition from microfluidic devices, which are somewhat specialized and challenging to use, towards low volume multiwell plates ubiquitous in biology laboratories. Using a combination of 3D printing and micromolding we have constructed inserts that can be placed into standard 12-well plates and can be used to create low volume culture conditions under which primary hepatocytes maintained a differentiated phenotype. This phenotype enhancement was confirmed by hepatic function assays including albumin synthesis and expression. Importantly we confirmed upregulation of HGF inside the low volume culture plates and demonstrated that inhibition of HGF signaling degraded the hepatic phenotype in our cell culture platform. Overall, this study outlines a new cell culture system that leverages the low volume effects of microfluidic channels in a multiwell plate format. Beyond hepatocytes, such a system may be of use in the maintenance of other difficult-to-culture cells including stem cells and primary cancer cells.


Why do some physicians in Portuguese-speaking African countries work exclusively for the private sector? Findings from a mixed-methods study.
Despite the growing interest in the private health sector in low- and middle-income countries, little is known about physicians working outside the public sector. The present work adopts a mixed-methods approach to explore characteristics, working patterns, choices, and motivations of the physicians working exclusively for the private sector in the capital cities of Cape Verde, Guinea Bissau, and Mozambique. The paper's objective is to contribute to the understanding of such physicians, ultimately informing the policies regulating the medical profession in low- and middle-income countries. The qualitative part of the study involved 48 interviews with physicians and health policy-makers and aimed at understanding the practice in the three locations. The quantitative study included a survey of 329 physicians, and multivariate analysis was conducted to analyse characteristics, time allocation, earnings, and motivations of those physicians working only for the private sector, in comparison to their public sector-only and dual practice peers. Our findings showed that only a limited proportion of physicians in the three locations work exclusively for the private sector (11.2%), with members of this group being older than those practicing only in the public or in both sectors. They were found to work fewer hours per week (49 hours) than their public (56 hours) and dual practice peers (62 hours) (P <0.001 and P = 0.011, respectively). Their median earnings were USD 4,405 per month, with substantial variations across the three locations. Statistically significant differences were found with the earnings of public-only physicians (P <0.001), but not with those of the dual practice group (P = 0.340). The qualitative data from the interviews showed private-only physicians' preference for an independent and more flexible work modality, and this was quoted as a determining factor for their choice of sector. This group appears to include those working in the more informal sector, and those who decided to leave the civil service following a disagreement with the public employer. The study shows the importance of understanding the relation between health professionals' characteristics, motivations, and their engagement with the private sector to develop effective policies to regulate the profession. This may ultimately contribute to achieving universal access to medical services in low- and middle-income countries.


A molecular portrait of maternal sepsis from Byzantine Troy.
Pregnancy complications are poorly represented in the archeological record, despite their importance in contemporary and ancient societies. While excavating a Byzantine cemetery in Troy, we discovered calcified abscesses among a woman's remains. Scanning electron microscopy of the tissue revealed 'ghost cells', resulting from dystrophic calcification, which preserved ancient maternal, fetal and bacterial DNA of a severe infection, likely chorioamnionitis. Gardnerella vaginalis and Staphylococcus saprophyticus dominated the abscesses. Phylogenomic analyses of ancient, historical, and contemporary data showed that G. vaginalis Troy fell within contemporary genetic diversity, whereas S. saprophyticus Troy belongs to a lineage that does not appear to be commonly associated with human disease today. We speculate that the ecology of S. saprophyticus infection may have differed in the ancient world as a result of close contacts between humans and domesticated animals. These results highlight the complex and dynamic interactions with our microbial milieu that underlie severe maternal infections.


Cyanobacterial diversity held in microbial biological resource centers as a biotechnological asset: the case study of the newly established LEGE culture collection.
Cyanobacteria are a well-known source of bioproducts which renders culturable strains a valuable resource for biotechnology purposes. We describe here the establishment of a cyanobacterial culture collection (CC) and present the first version of the strain catalog and its online database (http://lege.ciimar.up.pt/). The LEGE CC holds 386 strains, mainly collected in coastal (48%), estuarine (11%), and fresh (34%) water bodies, for the most part from Portugal (84%). By following the most recent taxonomic classification, LEGE CC strains were classified into at least 46 genera from six orders (41% belong to the Synechococcales), several of them are unique among the phylogenetic diversity of the cyanobacteria. For all strains, primary data were obtained and secondary data were surveyed and reviewed, which can be reached through the strain sheets either in the catalog or in the online database. An overview on the notable biodiversity of LEGE CC strains is showcased, including a searchable phylogenetic tree and images for all strains. With this work, 80% of the LEGE CC strains have now their 16S rRNA gene sequences deposited in GenBank. Also, based in primary data, it is demonstrated that several LEGE CC strains are a promising source of extracellular polymeric substances (EPS). Through a review of previously published data, it is exposed that LEGE CC strains have the potential or actual capacity to produce a variety of biotechnologically interesting compounds, including common cyanotoxins or unprecedented bioactive molecules. Phylogenetic diversity of LEGE CC strains does not entirely reflect chemodiversity. Further bioprospecting should, therefore, account for strain specificity of the valuable cyanobacterial holdings of LEGE CC.


Psoriasis and psoriatic arthritis video project: an update from the GRAPPA 2011 annual meeting.
Numerous physical examination instruments are used to assess and measure severity of psoriasis and psoriatic arthritis (PsA) in practice and in clinical trials. The Group for Research and Assessment of Psoriasis and Psoriatic Arthritis (GRAPPA) has developed several online training modules used by GRAPPA members and investigators participating in psoriasis and PsA research. At the 2011 GRAPPA meeting, attendees were updated on the ongoing development of the training modules. Several Internet-based multimedia presentations for psoriasis and PsA assessments have been completed. Available psoriasis modules include the Psoriasis Area and Severity Index (PASI) and Body Surface Area, one 5-point and two 6-point Physician Global Assessments, the original and modified Nail Psoriasis Severity Index, the Palmar-Plantar Pustular Psoriasis Area and Severity Index, the Psoriasis Scalp Severity Index, and the Total Plaque Severity Score. Rheumatology modules that demonstrate evaluation of swollen and tender joints, enthesitis, and dactylitis are now available, and an axial disease evaluation module is near completion. Each video includes the background and rationale for each measure, demonstration videos of select examinations, diagrams, and photographs to emphasize teaching points, and for most dermatology modules, an optional test to assess competence. Preliminary data generated by a pilot study of pre- and post-education PASI scoring by experienced and naive physicians and patient assessors were presented, revealing improved accuracy of scoring after viewing the PASI video. Attendees agreed that additional patient examples with more diverse skin types and psoriasis phenotypes, translation to languages other than English, and further validation studies are needed.


Role and involvement of life end information forum physicians in euthanasia and other end-of-life care decisions in Flanders, Belgium.
To describe role and involvement of Life End Information Forum (LEIF) physicians in end-of-life care decisions and euthanasia in Flanders. All 132 LEIF physicians in Belgium received a questionnaire inquiring about their activities in the past year, and their end-of-life care training and experience. Response rate was 75 percent. Most respondents followed substantive training in end-of-life care. In 1 year, LEIF physicians were contacted 612 times for consultations in end-of-life decisions, of which 355 concerned euthanasia requests eventually resulting in 221 euthanasia cases. LEIF physicians also gave information about various end-of-life issues (including palliative care) to patients and colleagues. LEIF physicians provide a forum for information and advice for physicians and patients. A similar health service providing support to physicians for all end-of-life decisions could also be beneficial for countries without a euthanasia law.


Effect of estrogen plus progestin on global cognitive function in postmenopausal women: the Women's Health Initiative Memory Study: a randomized controlled trial.
Observational studies have suggested that postmenopausal hormone treatment may improve cognitive function, but data from randomized clinical trials have been sparse and inconclusive. The Women's Health Initiative Memory Study (WHIMS) is an ancillary study of the Women's Health Initiative (WHI) hormone therapy trials. On July 8, 2002, the estrogen plus progestin therapy in the WHI trial was discontinued because of certain increased health risks for women. To determine whether estrogen plus progestin therapy protects global cognitive function in older postmenopausal women. A randomized, double-blind, placebo-controlled clinical trial, WHIMS is an ancillary study of geographically diverse, community-dwelling women aged 65 years or older from 39 of 40 clinical centers within the WHI estrogen plus progestin trial that started in June 1995. Of 4894 eligible postmenopausal women aged 65 years or older and free of probable dementia at baseline, 4532 (92.6%) were enrolled in the estrogen plus progestin component of WHIMS. A total of 4381 participants (96.7%) provided at least 1 valid cognitive function score between June 1995 and July 8, 2002. Participants received either 1 daily tablet containing 0.625 mg of conjugated equine estrogen with 2.5 mg of medroxyprogesterone acetate (n = 2145) or matching placebo (n = 2236). Global cognitive function measured annually with the Modified Mini-Mental State Examination. The Modified Mini-Mental State Examination mean total scores in both groups increased slightly over time (mean follow-up of 4.2 years). Women in the estrogen plus progestin group had smaller average increases in total scores compared with women receiving placebo (P =.03), but these differences were not clinically important. Removing women by censoring them after adjudicated dementia, mild cognitive impairment, or stroke, and nonadherence to study protocol, did not alter the findings. Prior hormone therapy use and duration of prior use did not affect the interpretation of the results, nor did timing of prior hormone therapy initiation with respect to the final menstrual period. More women in the estrogen plus progestin group had a substantial and clinically important decline (> or =2 SDs) in Modified Mini-Mental State Examination total score (6.7%) compared with the placebo group (4.8%) (P =.008). Among postmenopausal women aged 65 years or older, estrogen plus progestin did not improve cognitive function when compared with placebo. While most women receiving estrogen plus progestin did not experience clinically relevant adverse effects on cognition compared with placebo, a small increased risk of clinically meaningful cognitive decline occurred in the estrogen plus progestin group.


Disruption of neuropsin mRNA expression via RNA interference facilitates the photoinduced increase in thyrotropin-stimulating subunit β in birds.
It has long been known that the avian brain is capable of light detection independently of the eyes. The photoreceptive molecule neuropsin (OPN5) was identified in mammalian and avian brains, and shown to respond to biologically relevant light wavelengths. Whether OPN5 is functionally involved in light detection is unknown. Daylength plays a critical role in regulating the neuroendocrine control of reproduction in birds. The presence of light during a 'photoinducible' phase of the circadian cycle, which occurs 12-16 h after dawn, results in marked changes in hypothalamic gene expression. These changes ultimately control gonadotropin release from the pituitary gland that, in turn, stimulates gonadal development. In this study, we first measured OPN5 expression in the mediobasal hypothalamus (MBH) in border canaries during the photoinducible period in relation to thyrotropin (TSH) β-subunit mRNA expression, which is implicated in the control of avian reproduction. Second, the knockdown of OPN5 via small interfering RNA antisense in the MBH revealed that there is an inhibitory input in the photoinduced regulation of TSHβ mRNA expression. Our data indicate that a decrease in OPN5 mRNA expression is associated with the facilitation in TSHβ mRNA expression in the MBH, a critical step for the light-induced increase in gonadal recrudescence. We hypothesise that the removal of an inhibitory input by OPN5 in birds may be a step that occurs during the photoinducible period. Given the distribution of OPN5 in the brain and periphery, this suggests a possible multifunctional role for light information in regulating other physiological processes.


Adoptive T-cell transfer and chemotherapy in the first-line treatment of metastatic and/or locally recurrent nasopharyngeal carcinoma.
The outcomes for patients with metastatic or locally recurrent Epstein-Barr virus (EBV)-positive nasopharyngeal carcinoma (NPC) remain poor. Adoptive immunotherapy with EBV-specific cytotoxic T lymphocytes (EBV-CTLs) has proven clinical efficacy, but it has never been evaluated in the first-line treatment setting in combination with chemotherapy. To evaluate the safety and efficacy of a chemotherapy in combination with adoptive EBV-CTL transfer, we conducted a phase 2 clinical trial consisting of four cycles of gemcitabine and carboplatin (GC) followed by up to six doses of EBV-CTL. Thirty-eight patients were enrolled, and 35 received GC and EBV-CTL. GC-CTL therapy resulted in a response rate of 71.4% with 3 complete responses and 22 partial responses. With a median follow up of 29.9 months, the 2-year and 3-year overall survival (OS) rate was 62.9 and 37.1%, respectively. Five patients did not require further chemotherapy for more than 34 months since initiation of CTL. Infusion of CTL products containing T cells specific for LMP2 positively correlated with OS (hazard ratio: 0.35; 95% confidence interval: 0.14-0.84; P = 0.014). Our study achieved one of the best survival outcomes in patients with advanced NPC, setting the stage for a future randomized study of chemotherapy with and without EBV-CTL.


Ancient Lowland Maya neighborhoods: Average Nearest Neighbor analysis and kernel density models, environments, and urban scale.
Many humans live in large, complex political centers, composed of multi-scalar communities including neighborhoods and districts. Both today and in the past, neighborhoods form a fundamental part of cities and are defined by their spatial, architectural, and material elements. Neighborhoods existed in ancient centers of various scales, and multiple methods have been employed to identify ancient neighborhoods in archaeological contexts. However, the use of different methods for neighborhood identification within the same spatiotemporal setting results in challenges for comparisons within and between ancient societies. Here, we focus on using a single method-combining Average Nearest Neighbor (ANN) and Kernel Density (KD) analyses of household groups-to identify potential neighborhoods based on clusters of households at 23 ancient centers across the Maya Lowlands. While a one-size-fits all model does not work for neighborhood identification everywhere, the ANN/KD method provides quantifiable data on the clustering of ancient households, which can be linked to environmental zones and urban scale. We found that centers in river valleys exhibited greater household clustering compared to centers in upland and escarpment environments. Settlement patterns on flat plains were more dispersed, with little discrete spatial clustering of households. Furthermore, we categorized the ancient Maya centers into discrete urban scales, finding that larger centers had greater variation in household spacing compared to medium-sized and smaller centers. Many larger political centers possess heterogeneity in household clustering between their civic-ceremonial cores, immediate hinterlands, and far peripheries. Smaller centers exhibit greater household clustering compared to larger ones. This paper quantitatively assesses household clustering among nearly two dozen centers across the Maya Lowlands, linking environment and urban scale to settlement patterns. The findings are applicable to ancient societies and modern cities alike; understanding how humans form multi-scalar social groupings, such as neighborhoods, is fundamental to human experience and social organization.


High-frequency demodulation of multiphoton fluorescence in long-wavelength hyper-Rayleigh scattering.
Suppression of the multiphoton fluorescence contribution to the hyper-Rayleigh (second-order nonlinear optical) scattering signal was recently achieved by intrinsic demodulation of the fluorescence at high amplitude-modulation (AM) frequencies [Olbrechts et al., Rev. Sci. Instrum. 69, 2233 (1998)]. These high AM frequencies were obtained from the high harmonic content in the Fourier spectrum of a repetitive train of femtosecond pulses from a Ti:sapphire laser emitting at 800 nm. We have used a femtosecond parametric oscillator to shift the fundamental wavelength to 1.3 mum . By further improving the detection electronics, we can now obtain fluorescence suppression at AM frequencies up to 600 MHz. Fluorescence-free hyperpolarizability values were obtained for fluorescent dipolar compounds as well as for an ionic fluorophore. The results also indicate that shifting the fundamental wavelength to the near infrared only is not a general solution to the multiphoton fluorescence problem in hyper-Rayleigh scattering.


Non-modulation as an intermediate phenotype in essential hypertension.
Non-modulation is a trait characterized by abnormal angiotensin-mediated control of aldosterone release and the renal blood supply. To determine whether non-modulation defines a specific subgroup of the hypertensive population and its utility as an intermediate phenotype, we have studied the distribution of this quantitative trait, whether its features are reproducible on repeated testing, and whether there is concordance of its multiple features. Essential hypertensive patients (224) and normotensive subjects (119) received an infusion of angiotensin II (Ang II) at 3 ng.kg-1.min-1 for 30-45 minutes. p-Aminohippurate (PAH) clearance was assessed as an index of renal plasma flow while the subjects were on a 200 meq sodium diet; plasma aldosterone levels were measured while the subjects were on a 10 meq sodium diet. In 54 subjects, diuretic-induced volume depletion superimposed on a low salt diet was substituted for the Ang II infusion. The results of each study were submitted to maximum likelihood analysis to assess bimodality. In response to both diuretic-induced volume depletion (p < 0.000023) and Ang II infusion (p < 0.0009), aldosterone responses were bimodally distributed in the essential hypertensive but not in the normotensive subjects, suggesting that this trait identifies a discrete subgroup. In the 59 subjects who had both an adrenal and renal study, 50 (85%) were concordant. Finally, in 27 subjects studied two to six times over a span of 1-60 months, the intraclass correlations of the adrenal, PAH, or both responses were highly significant (p values between 0.001 and 0.00007), indicating high reproducibility of results on repeated testing.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of the T-domain on intracellular transport of diphtheria toxin.
Subunit B of diphtheria toxin (DT), which consists of two domains: R (receptor-binding) and T (transmembrane), plays an important role in toxin-receptor binding on the cell-targets and in transportation of catalytic subunit A to the cell cytosol. Recombinant analogues of the subunit B are promising representatives in the unique class of transporting proteins, able to deliver different types of biologically active molecules to cell cytosol. In the development of these protein constructs understanding of the role of each of the DT fragments in determination of transporting pathways of endocytosed complex toxin-receptor is urgently required. We have studied in this work the T-domain effect on intracellular transport of recombinant fragments of DT. We have compared intracellular transport of the R-domain and the subunit B, the last one consisted of both R-domain and T-domain. Recombinant fragments of DT used in this work were labeled with fluorescent proteins, which allowed applying colocalization technique for our study. Application of confocal microscopy technique revealed differences in transportation of recombinant derivates of DT in Vero cells: R-domain moved faster than subunit B to tubular compartments. Analysis of R-domain and subunit B transportation confirmed almost linear increase of their colocalization with the time regarding to Pearsons correlation coefficient (PCC). However, amount of colocalized with R-domain subunit B were not linearly increased with time according to Manders coefficient (M1), this could indicate the ability of subunit B to transport to such compartments that R-domain do not reach. Possible role of the T-domain in intracellular transportation and compartmentalization of the toxin may be associated with the ability of the T-domain to form a proton channels and its ability to interact with COPI complex.


Loss of Stat6 affects chromatin condensation in intestinal epithelial cells causing diverse outcome in murine models of inflammation-associated and sporadic colon carcinogenesis.
While great advances have been achieved regarding the genetic basis of colorectal cancer, the complex role of cell-cell communication and cytokine-induced signaling during its pathogenesis remains less understood. Signal transducer and activator of transcription 6 (Stat6) is the main transcription factor of interleukin-4 (IL-4) signaling and its participation in the development of various tumor types has been already reported. Here we aimed to examine the contribution of Stat6 in intestinal epithelial cells (IEC) in mouse models of intestinal carcinogenesis. Wild-type (WT), Stat6 knockout (Stat6-/-), and intestinal epithelial cell-specific IL-4Rα knockout (Il-4rαΔIEC) mice were subjected to colitis-associated (AOM/DSS) and colitis-independent (sporadic) carcinogenesis. IEC proliferation, apoptosis and RNA expression were evaluated by immunohistochemical, immunoblot, and RT-PCR analysis. We found that Stat6-/- mice developed more tumors in the colitis-associated carcinogenesis model. This was accompanied by a more pronounced inflammatory response during colitis and an elevated Stat3-dependent proliferation of IEC. Increased sensitivity to DSS-induced colitis was caused by elevated cell death in response to the initial carcinogen exposure as Stat6 deficiency led to increased chromatin compaction affecting DNA damage response in IEC upon treatment with alkylating agents independently of IL-4Rα engagement. Thus, loss of Stat6 caused more severe colitis and increased tumor load, however loss-of-initiated Stat6-/- IEC prevented tumor formation in the absence of overt inflammation. Our data unravel unexpected IL-4-independent functions of Stat6 in chromatin compaction in intestinal epithelial cells ultimately providing both tumor suppressive as well as tumor promoting effects in different models of intestinal tumorigenesis.


Systematic review of the cost effectiveness of radiation therapy for prostate cancer from 2003 to 2013.
Prostate cancer remains a prevalent diagnosis with a spectrum of treatment choices that offer similar oncologic outcomes but differing side effect profiles and associated costs. As the technology for prostate radiation therapy has advanced, its associated costs have escalated, thus making cost-effectiveness analyses critical to assess the value of competing treatment options, including watchful waiting, surgery, brachytherapy, intensity-modulated radiation therapy (IMRT), 3D-conformal radiation therapy (3D-CRT), proton beam therapy (PBT), and stereotactic body radiation therapy (SBRT). The aim of this systematic review was to identify articles that performed a cost-effectiveness analysis on different radiation treatment options for localized prostate cancer, summarize their findings, and highlight the main drivers of cost effectiveness. A literature search was performed on two databases, PubMed and the Cost-Effectiveness Analysis Registry ( https://research.tufts-nemc.org/cear4 ), using search terms that included 'prostate', 'cost effectiveness prostate radiation' and 'cost analysis comparative effectiveness prostate radiation'. Studies were included in this review if the cost data were from 2002 or later, and outcomes reported both cost and effectiveness, preferably including a cost-utility analysis with the outcome of an incremental cost-effectiveness ratio with quality-adjusted life-year (QALY) as the effectiveness measure. There were 14 articles between 2003 and 2013 that discussed cost effectiveness of prostate radiotherapy in men over the age of 65. All but four of the papers were from the US; the others were from Canada and the UK. The majority of the papers used Markov decision analysis and estimated cost from a payer's perspective, usually from Medicare reimbursement data. Assumptions for the model and utilities to calculate QALYs were estimated using published literature at the time of the analysis. Each analysis had a sensitivity analysis to compensate for the uncertainty of the model inputs. The main drivers of cost effectiveness were the cost of the radiation treatment and the differential QALYs accrued because of different treatment-related morbidities. Brachytherapy was consistently found to be more cost effective when compared with surgery and other radiation treatment options. IMRT was cost effective when compared with 3D-CRT. PBT was not found to be cost effective in any of the analyses, mostly due to the high costs of PBT. SBRT was the newest technology that was analyzed, and it was also found to be cost effective compared with IMRT and PBT. Cost-effectiveness research of prostate radiation treatments allows patients, providers, and payers to better understand the true value of each treatment choice. Due to the variation in each of these analyses (e.g., costing, and disease and complication assumptions, etc.), it is difficult to generalize the results. One must be careful in drawing conclusions from these studies and extrapolating to individual patients, particularly with the clear utility dependence seen in the majority of these studies.


Hierarchies, similarity, and interactivity in object recognition: "category-specific" neuropsychological deficits.
Category-specific impairments of object recognition and naming are among the most intriguing disorders in neuropsychology, affecting the retrieval of knowledge about either living or nonliving things. They can give us insight into the nature of our representations of objects: Have we evolved different neural systems for recognizing different categories of object? What kinds of knowledge are important for recognizing particular objects? How does visual similarity within a category influence object recognition and representation? What is the nature of our semantic knowledge about different objects? We review the evidence on category-specific impairments, arguing that deficits even for one class of object (e.g., living things) cannot be accounted for in terms of a single information processing disorder across all patients; problems arise at contrasting loci in different patients. The same apparent pattern of impairment can be produced by damage to different loci. According to a new processing framework for object recognition and naming, the hierarchical interactive theory (HIT), we have a hierarchy of highly interactive stored representations. HIT explains the variety of patients in terms of (1) lesions at different levels of processing and (2) different forms of stored knowledge used both for particular tasks and for particular categories of object.


Repetitive locomotor training and physiotherapy improve walking and basic activities of daily living after stroke: a single-blind, randomized multicentre trial (DEutsche GAngtrainerStudie, DEGAS).
To evaluate the effect of repetitive locomotor training on an electromechanical gait trainer plus physiotherapy in subacute stroke patients. Randomized controlled trial. Four German neurological rehabilitation centres. One hundred and fifty-five non-ambulatory patients (first-time stroke <60 days). Group A received 20 min locomotor training and 25 min physiotherapy; group B had 45 min physiotherapy every week day for four weeks. Primary variables were gait ability (Functional Ambulation Category, 0-5) and the Barthel Index (0-100), blindly assessed at study onset, end, and six months later for follow-up. Responders to the therapy had to become ambulatory (Functional Ambulation Category 4 or 5) or reach a Barthel Index of > or = 75. Secondary variables were walking velocity, endurance, mobility and leg power. The intention-to-treat analysis revealed that significantly greater number of patients in group A could walk independently: 41 of 77 versus 17 of 78 in group B (P B < 0.0001) at treatment end. Also, significantly more group A patients had reached a Barthel Index > or = 75: 44 of 77 versus 21 of 78 (P B < 0.0001). At six-month follow-up, the superior gait ability in group A persisted (54 of 77 versus 28 of 78, P B < 0.0001), while the Barthel Index responder rate did not differ. For all secondary variables, group A patients had improved significantly more (P B < 0.0001) during the treatment period, but not during follow-up. Intensive locomotor training plus physiotherapy resulted in a significantly better gait ability and daily living competence in subacute stroke patients compared with physiotherapy alone.


Identification and localization of the multiple bacterial symbionts of the termite gut flagellate Joenia annectens.
The hindgut of wood-feeding lower termites is densely colonized by a multitude of symbiotic micro-organisms. While it is well established that the eukaryotic flagellates play a major role in the degradation of lignocellulose, much less is known about the identity and function of the prokaryotic symbionts associated with the flagellates. Our ultrastructural investigations of the gut flagellate Joenia annectens (from the termite Kalotermes flavicollis) revealed a dense colonization of this flagellate by diverse ecto- and endosymbiotic bacteria. Phylogenetic analysis of the small-subunit rRNA gene sequences combined with fluorescence in situ hybridization allowed us to identify and localize the different morphotypes. Furthermore, we could show that K. flavicollis harbours two phylotypes of J. annectens that could be distinguished not only by their small-subunit rRNA gene sequences, but also by differences in their assemblages of bacterial symbionts. Each of the flagellate populations hosted phylogenetically distinct ectosymbionts from the phylum Bacteroidetes, one of them closely related to the ectosymbionts of other termite gut flagellates. A single phylotype of 'Endomicrobia' was consistently associated with only one of the host phylotypes, although not all individuals were colonized, corroborating that 'Endomicrobia' symbionts do not always cospeciate with their host lineages. Flagellates from both populations were loosely associated with a single phylotype of Spirochaetales attached to their cell surface in varying abundance. Current evidence for the involvement of Bacteroidales and 'Endomicrobia' symbionts in the nitrogen metabolism of the host flagellate is discussed.


Balancing ballistic protection against physiological strain: evidence from laboratory and field trials.
This project was based on the premise that decisions concerning the ballistic protection provided to defence personnel should derive from an evaluation of the balance between protection level and its impact on physiological function, mobility, and operational capability. Civilians and soldiers participated in laboratory- and field-based studies in which ensembles providing five levels of ballistic protection were evaluated, each with progressive increases in protection, mass (3.4-11.0 kg), and surface-area coverage (0.25-0.52 m(2)). Physiological trials were conducted on volunteers (N = 8) in a laboratory, under hot-dry conditions simulating an urban patrol: walking at 4 km·h(-1) (90 min) and 6 km·h(-1) (30 min or to fatigue). Field-based trials were used to evaluate tactical battlefield movements (mobility) of soldiers (N = 31) under tropical conditions, and across functional tests of power, speed, agility, endurance, and balance. Finally, trials were conducted at a jungle training centre, with soldiers (N = 32) patrolling under tropical conditions (averaging 5 h). In the laboratory, work tolerance was reduced as protection increased, with deep-body temperature climbing relentlessly. However, the protective ensembles could be grouped into two equally stressful categories, each providing a different level of ballistic protection. This outcome was supported during the mobility trials, with the greatest performance decrement evident during fire and movement simulations, as the ensemble mass was increased (-2.12%·kg(-1)). The jungle patrol trials similarly supported this outcome. Therefore, although ballistic protection does increase physiological strain, this research has provided a basis on which to determine how that strain can be balanced against the mission-specific level of required personal protection.


dMyc transforms cells into super-competitors.
Overexpression of myc protooncogenes has been implicated in the genesis of many human tumors. Myc proteins seem to regulate diverse biological processes, but their role in tumorigenesis remains enigmatic. Here we use Drosophila imaginal discs to mimic situations in which cells with unequal levels of Myc protein are apposed and show that this invariably elicits a win/lose situation reminiscent of cell competition; cells with lower levels of dMyc are eliminated by apoptosis whereas cells with higher levels of dMyc over-proliferate. We find that this competitive behavior correlates with, and can be corrected by, the activation of the BMP/Dpp survival signaling pathway. Hence the heritable increase in dMyc levels causes cells to behave as "super-competitors" and reveals a novel mode of clonal expansion that causes, but also relies on, the killing of surrounding cells.


Comparison of hepatitis B virus subtyping of d/y determinants by radioimmunoprecipitation assay and the polymerase chain reaction.
Using a double polymerase chain reaction a method was devised for detecting and subtyping hepatitis B virus DNA in serum samples. Primers from the S-gene were selected from the sequence analyses of five HBV HBsAg subtypes, to amplify HBV DNA and subtype for y specific DNA. Thirty-eight samples were subtyped for d and y determinants by radioimmunoprecipitation assay (RIPA) and the polymerase chain reaction (PCR). Subtyping by PCR and RIPA was in agreement in 100% of subtype y samples and 83.3% of subtype d, giving an overall correlation of 92.1%. As a third comparison, 12 amplified samples were digested by the restriction enzyme Sau 3A, which differentiates between subtypes y and d. The digest results agreed with PCR in 83.3% of the samples. In addition, we compared our standard phenol/chloroform extraction against a rapid one step method. The phenol/chloroform stage was found to be essential for the removal of nucleases and polymerase inhibitors present in sera.


New therapy using omega-3-Acid ethyl esters for decubitus ulcers and stasis dermatitis: a case report.
In daily practice, it is common to experience difficulty in treating decubitus ulcers (pressure ulcers, also known as decubitus ulcers) and stasis dermatitis of the lower limbs. We hereby report that omega-3-acid ethyl esters were remarkably effective when administered to cases of refractory pressure ulcers and stasis dermatitis for the purpose of improving the blood flow and promoting blood circulation. Case 1: A 21-year-old Japanese female with lower-body paralysis. Pressure ulcers appeared on the heel and first toe of her left lower extremity. Although the patient had been treated with various ointments such as dimethyl isopropylazulene and 0.9% iodine-containing ointment, the course showed no improvement, so omega-3-acid ethyl esters was administered orally, completely healing the ulcer of the first toe in 10 weeks. Case 2: A 76-year-old Japanese male. The patient had been treated on an outpatient basis for 15 years due to hypertension, heart failure, type 2 diabetes mellitus, and hyperlipidemia. Two years prior to this presentation, stasis dermatitis occurred in the lower limbs and at the end of last year, erosive ulcers appeared on the front part of the lower-right thigh and shin. Although treatment with various topical ointment and dressings was performed, the course showed no improvement. Oral administration of omega-3-acid ethyl esters was initiated. At 12 weeks, his condition entered the white phase and healed almost completely. This report is the first to document other treatment possibilities for pressure ulcer and/or stasis dermatitis in cases where the use of topical applied ointments and medications is difficult. This new therapy may therefore help physicians to treat pressure ulcers and stasis dermatitis.


Safer sex media messages and adolescent sexual behavior: 3-year follow-up results from project iMPPACS.
We estimated the long-term (36-month) effects of Project iMPPACS, a multisite randomized controlled trial of mass media and small-group intervention for African American adolescents. We collected 6 waves of longitudinal data on program participants aged 14 to 17 years (n = 1139) in Providence, Rhode Island; Syracuse, New York; Columbia, South Carolina; and Macon, Georgia, 36 months (December 2009-December 2010) after the intervention began (August 2006-January 2008). Seemingly unrelated regressions at each wave estimated the effects of 3 types of mass media messages (the thematic mediators: selection, pleasure, and negotiation) on condom use intention and self-reported unprotected vaginal sex events. All 3 mediators of behavior change that were introduced during the media intervention were sustained at the follow-up assessments at least 18 months after the intervention ended, with intention having the largest correlation. Unprotected vaginal sex increased with each wave of the study, although cities receiving media exposure had smaller increases. Project iMPPACS demonstrates that mass media influence delivered over an extended period, when adolescents were beginning to learn patterns of behavior associated with sex, persisted after the media program ended.


Oxytocin, Vasopressin and Prolactin in New Breastfeeding Mothers: Relationship to Clinical Characteristics and Infant Weight Loss.
Maternal milk production requires the neuropeptide oxytocin. Individual variation in oxytocin function is a compelling target for understanding low milk production, a leading cause of breastfeeding attrition. Complicating the understanding of oxytocin pathways is that vasopressin may interact with oxytocin receptors, yet little is known about the role of vasopressin in lactation. The aims of this study were (1) to describe maternal plasma oxytocin, vasopressin, and prolactin patterns during breastfeeding following low-risk spontaneous labor and birth in healthy first-time mothers and (2) to relate hormone patterns to maternal characteristics and breastfeeding measures. Eligible women were recruited before hospital discharge. Forty-six participants enrolled and 35 attended the study visit. Participants kept a journal of breastfeeding frequency, symptoms of lactogenesis, and infant weight. Plasma samples were obtained at breastfeeding onset on Day 4-5 postpartum, and repeated after 20 min. Hormones were measured with immunoassays. Infant weight change, milk transfer, and onset of lactogenesis were also measured. Baseline oxytocin and vasopressin were inversely related to one another. Oxytocin and prolactin increased significantly across the 20-min sampling period while vasopressin decreased. Higher oxytocin was associated with higher maternal age, lower BMI, shorter active labor, physiologic labor progression, and less weight loss in the newborn. Higher vasopressin correlated with younger maternal age, higher BMI, and greater newborn weight loss. Oxytocin and vasopressin have contrasting relationships with maternal clinical characteristics and newborn weight gain in early breastfeeding infants. Further study is needed to understand how oxytocin and vasopressin influence lactation outcomes.