Evaluation of infradiaphragmatic total anomalous pulmonary venous connection with two-dimensional echocardiography.
In three newborn infants with infradiaphragmatic total anomalous pulmonary venous connection, the common pulmonary vein was visualized with two-dimensional echocardiography and validated with saline contrast injections. The transducer was placed in the subcostal region in a sagittal body plane so as to image the descending aorta and the vertebral column. The common pulmonary vein, which connected the pulmonary veins to a systemic vein in the abdomen, was seen lying parallel and anterior to the descending aorta and to the left of the inferior vena cava. With peripheral venous contrast injections, contrast echoes first filled the inferior vena cava and then the descending aorta because of obligatory right-to-left atrial shunting. The common pulmonary vein was the only structure that remained free of contrast echoes. Thus, contrast echocardiography provided a method for identifying the site of pulmonary venous drainage and for distinguishing the common pulmonary vein from other structures such as the inferior vena cava. In cyanotic infants with respiratory distress, two-dimensional contrast echocardiography permits a rapid diagnosis of infradiaphragmatic total anomalous pulmonary venous connection.


Effects of short-term nicotine deprivation on decision-making: delay, uncertainty and effort discounting.
Experimental evidence suggests that when opioid-dependent drug users are deprived of heroin, they become more likely to behave impulsively on a computer task. The current study examined whether nicotine deprivation has similar effects in cigarette smokers, causing an increase in impulsive decision-making. Simultaneously, the impact of deprivation on several other related decision-making tasks was assessed. Eleven smokers (> or = 15 cigarettes/day) participated in two experimental sessions. For one session, they smoked as usual until the session began. For the other, participants did not smoke for 24 hr. Abstinence was verified using levels of breath carbon monoxide and urinary cotinine. During each session, they completed computer tasks that assessed impulsivity by measuring the tendency to choose small, immediate rewards (cigarettes or money) over US dollar 10 available following a delay, as well as tasks in which smokers chose between small, certain, easily obtained rewards and US dollar 10 whose availability was uncertain or required high levels of effort to obtain. Deprivation increased preference for immediate cigarettes over delayed money, but did not alter preference for immediate money over delayed money. These findings indicate that short-term nicotine abstinence does increase impulsive decision-making, but only when the impulsive choice is drug-related. Increases were not related to a general increase in the value of immediate rewards per se or a general increase in aversion to delayed rewards. Decision-making in the other tasks followed a similar pattern: Deprivation increased preference for the cigarette alternative but did not alter the decision-making processes for nondrug rewards.


Hydrogen production by termite gut protists: characterization of iron hydrogenases of Parabasalian symbionts of the termite Coptotermes formosanus.
Cellulolytic flagellated protists in the guts of termites produce molecular hydrogen (H(2)) that is emitted by the termites; however, little is known about the physiology and biochemistry of H(2) production from cellulose in the gut symbiotic protists due to their formidable unculturability. In order to understand the molecular basis for H(2) production, we here identified two genes encoding proteins homologous to iron-only hydrogenases (Fe hydrogenases) in Pseudotrichonympha grassii, a large cellulolytic symbiont in the phylum Parabasalia, in the gut of the termite Coptotermes formosanus. The two Fe hydrogenases were phylogenetically distinct and had different N-terminal accessory domains. The long-form protein represented a phylogenetic lineage unique among eukaryotic Fe hydrogenases, whereas the short form was monophyletic with those of other parabasalids. Active recombinant enzyme forms of these two Fe hydrogenases were successfully obtained without the specific auxiliary maturases. Although they differed in their extent of specific activity and optimal pH, both enzymes preferentially catalyzed H(2) evolution rather than H(2) uptake. H(2) evolution, at least that associated with the short-form enzyme, was still active even under high hydrogen partial pressure. H(2) evolution activity was detected in the hydrogenosomal fraction of P. grassii cells; however, the vigorous H(2) uptake activity of the endosymbiotic bacteria compensated for the strong H(2) evolution activity of the host protists. The results suggest that termite gut symbionts are a rich reservoir of novel Fe hydrogenases whose properties are adapted to the gut environment and that the potential of H(2) production in termite guts has been largely underestimated.


Mechanism of the central effects of dopamine and metoclopramide on aldosterone regulation in the rat.
To investigate the mechanism of the central action of dopamine and its antagonist, metoclopramide, on the regulation of aldosterone, studies were performed in 54 conscious rats with and without bilateral nephrectomy. In normal and sham-operated rats, intracerebroventricular injection of dopamine resulted in a significant suppression of plasma renin activity and plasma aldosterone at 30 min, and intracerebroventricular injection of metoclopramide resulted in a significant elevation of plasma renin activity and plasma aldosterone at 30 min without altering the plasma corticosterone and potassium levels. In bilaterally nephrectomized rats, the plasma renin activity was significantly reduced and it did not respond to dopamine or metoclopramide. In these rats, intracerebroventricular injection of metoclopramide exerted no effect on the plasma aldosterone, but intracerebroventricular injection of dopamine increased the plasma aldosterone slightly. However, this increase was not statistically significant. These findings suggest that the dopaminergic system in the brain is involved in the regulation of aldosterone secretion, mainly with changes in the peripheral renin-angiotensin axis in rats.


Meerkat close calling patterns are linked to sex, social category, season and wind, but not fecal glucocorticoid metabolite concentrations.
It is well established that animal vocalizations can encode information regarding a sender's identity, sex, age, body size, social rank and group membership. However, the association between physiological parameters, particularly stress hormone levels, and vocal behavior is still not well understood. The cooperatively breeding African meerkats (Suricata suricatta) live in family groups with despotic social hierarchies. During foraging, individuals emit close calls that help maintain group cohesion. These contact calls are acoustically distinctive and variable in rate across individuals, yet, information on which factors influence close calling behavior is missing. The aim of this study was to identify proximate factors that influence variation in call rate and acoustic structure of meerkat close calls. Specifically, we investigated whether close calling behavior is associated with sex, age and rank, or stress hormone output (i.e., measured as fecal glucocorticoid metabolite (fGCM) concentrations) as individual traits of the caller, as well as with environmental conditions (weather) and reproductive seasonality. To disentangle the effects of these factors on vocal behavior, we analyzed sound recordings and assessed fGCM concentrations in 64 wild but habituated meerkats from 9 groups during the reproductive and non-reproductive seasons. Dominant females and one-year old males called at significantly higher rates compared to other social categories during the reproductive season. Additionally, dominant females produced close calls with the lowest mean fundamental frequencies (F0) and the longest mean pulse durations. Windy conditions were associated with significantly higher call rates during the non-reproductive season. Fecal GCM concentrations were unrelated to close calling behavior. Our findings suggest that meerkat close calling behavior conveys information regarding the sex and social category of the caller, but shows no association with fGCM concentrations. The change in call rate in response to variation in the social and ecological environments individuals experience indicates some degree of flexibility in vocal production.


SpMyb functions as an intramodular repressor to regulate spatial expression of CyIIIa in sea urchin embryos.
The CyIIIa actin gene of Strongylocentrotus purpuratus is transcribed exclusively in the embryonic aboral ectoderm, under the control of 2.3 kb cis-regulatory domain that contains a proximal module that controls expression in early embryogenesis, and a middle module that controls expression in later embryogenesis. Previous studies demonstrated that the SpRunt-1 target site within the middle module is required for the sharp increase in CyIIIa transcription which accompanies differentiation of the aboral ectoderm, and that a negative regulatory region near the SpRunt-1 target site is required to prevent ectopic transcription in the oral ectoderm and skeletogenic mesenchyme. This negative regulatory region contains a consensus binding site for the myb family of transcription factors. In vitro DNA-binding experiments reveal that a protein in blastula-stage nuclei interacts specifically with the myb target site. Gene transfer experiments utilizing CyIIIa reporter constructs containing oligonucleotide substitutions indicate that this site is both necessary and sufficient to prevent ectopic expression of CyIIIa. Synthetic oligonucleotides containing the myb target site were used to purify a protein from sea urchin embryo nuclear extracts by affinity chromatography. This protein is immunoprecipitated by antibodies specific to the evolutionarily conserved myb domain, and amino acid sequences obtained from the purified protein were found to be identical to sequences within the myb domain. Sequence information was used to obtain cDNA clones of SpMyb, the S. purpuratus member of the myb family of transcription factors. Through interactions within the middle module, SpMyb functions to repress activation of CyIIIa in the oral ectoderm and skeletogenic mesenchyme.


Differentiating midazolam over-sedation from neurological damage in the intensive care unit.
Midazolam is used routinely to sedate patients in the intensive care unit (ICU). We suspected that midazolam over-sedation was occurring in the ICU of the Guy's and St. Thomas' Trust and that it could be difficult to differentiate this from underlying neurological damage. A sensitive assay for detecting midazolam and 1-hydroxymidazolam glucuronide (1-OHMG) in serum was developed and applied in the clinical setting. In the present study we evaluated a series of cases managed in a mixed medical, surgical and trauma ICU. Serum was collected from 26 patients who received midazolam, were 'slow to wake' and in whom there was suspicion of neurological damage. Patient outcome was followed in terms of mortality, neurological recovery and neurological damage on discharge. Out of 26 patients, 13 had detectable serum levels of midazolam and/or 1-OHMG after a median of 67 hours (range 36-146 hours) from midazolam cessation. Of these 13 patients in whom midazolam/1-OHMG was detectable, 10 made a full neurological recovery. Of the remaining 13 patients with no detectable midazolam/1-OHMG, three made a full neurological recovery; 10 patients were subsequently found to have suffered neurological damage (P < 0.002), eight of whom died and two were discharged from the ICU with profound neurological damage. These findings confirm that prolonged sedation after midazolam therapy should be considered in the differential diagnosis of neurological damage in the ICU. This can be reliably detected by the assay method described. The effects of midazolam/1-OHMG persist days after administration of midazolam has ceased. After prolonged sedation has been excluded in this patient group, it is highly likely that neurological damage has occurred.


Safety and immunogenicity of an AMA1 malaria vaccine in Malian children: results of a phase 1 randomized controlled trial.
The objective was to evaluate the safety and immunogenicity of the AMA1-based malaria vaccine FMP2.1/AS02(A) in children exposed to seasonal falciparum malaria. A Phase 1 double blind randomized controlled dose escalation trial was conducted in Bandiagara, Mali, West Africa, a rural town with intense seasonal transmission of Plasmodium falciparum malaria. The malaria vaccine FMP2.1/AS02(A) is a recombinant protein (FMP2.1) based on apical membrane antigen 1 (AMA1) from the 3D7 clone of P. falciparum, formulated in the Adjuvant System AS02(A). The comparator vaccine was a cell-culture rabies virus vaccine (RabAvert). One hundred healthy Malian children aged 1-6 years were recruited into 3 cohorts and randomized to receive either 10 microg FMP2.1 in 0.1 mL AS02(A), or 25 microg FMP2.1 in 0.25 mL AS02(A), or 50 microg FMP2.1 50 microg in 0.5 mL AS02(A), or rabies vaccine. Three doses of vaccine were given at 0, 1 and 2 months, and children were followed for 1 year. Solicited symptoms were assessed for 7 days and unsolicited symptoms for 30 days after each vaccination. Serious adverse events were assessed throughout the study. Transient local pain and swelling were common and more frequent in all malaria vaccine dosage groups than in the comparator group, but were acceptable to parents of participants. Levels of anti-AMA1 antibodies measured by ELISA increased significantly (at least 100-fold compared to baseline) in all 3 malaria vaccine groups, and remained high during the year of follow up. The FMP2.1/AS02(A) vaccine had a good safety profile, was well-tolerated, and induced high and sustained antibody levels in malaria-exposed children. This malaria vaccine is being evaluated in a Phase 2 efficacy trial in children at this site. ClinicalTrials.gov NCT00358332 [NCT00358332].


Uncovering the complexity of transcriptomes with RNA-Seq.
In recent years, the introduction of massively parallel sequencing platforms for Next Generation Sequencing (NGS) protocols, able to simultaneously sequence hundred thousand DNA fragments, dramatically changed the landscape of the genetics studies. RNA-Seq for transcriptome studies, Chip-Seq for DNA-proteins interaction, CNV-Seq for large genome nucleotide variations are only some of the intriguing new applications supported by these innovative platforms. Among them RNA-Seq is perhaps the most complex NGS application. Expression levels of specific genes, differential splicing, allele-specific expression of transcripts can be accurately determined by RNA-Seq experiments to address many biological-related issues. All these attributes are not readily achievable from previously widespread hybridization-based or tag sequence-based approaches. However, the unprecedented level of sensitivity and the large amount of available data produced by NGS platforms provide clear advantages as well as new challenges and issues. This technology brings the great power to make several new biological observations and discoveries, it also requires a considerable effort in the development of new bioinformatics tools to deal with these massive data files. The paper aims to give a survey of the RNA-Seq methodology, particularly focusing on the challenges that this application presents both from a biological and a bioinformatics point of view.


Clinical management of an unusual case of gingival enlargement.
The purpose of this article is to report a case of conditioned gingival enlargement managed by non-surgical periodontal therapy. Hormones are specific regulatory molecules that modulate a host of body functions. Hormonal effects reflect physiologic and pathologic changes in almost all tissues of the body with the periodontium being no exception. Physiologic changes like puberty, the menstrual cycle, and pregnancy cause hormonal variations that may cause inflammation of the gingiva. Oral contraceptives that contain estrogen and/or progesterone are associated with gingival enlargement. A 28-year-old female presented with a complaint of swelling of the gingiva with spontaneous bleeding in the maxillary anterior region for a period of one year. The health history documented the use of contraceptives for one year, and a clinical examination revealed the existence of poor oral hygiene and enlarged painful gingival tissues that bled when touched. This case reaffirms the fact plaque control is the most important procedure in any periodontal therapy. Another factor contributing to the excellent response to therapy is patient compliance. The patient followed home care instructions well and was effective in personal oral hygiene measures.


Citizen Science for Mining the Biomedical Literature.
Biomedical literature represents one of the largest and fastest growing collections of unstructured biomedical knowledge. Finding critical information buried in the literature can be challenging. To extract information from free-flowing text, researchers need to: 1. identify the entities in the text (named entity recognition), 2. apply a standardized vocabulary to these entities (normalization), and 3. identify how entities in the text are related to one another (relationship extraction). Researchers have primarily approached these information extraction tasks through manual expert curation and computational methods. We have previously demonstrated that named entity recognition (NER) tasks can be crowdsourced to a group of non-experts via the paid microtask platform, Amazon Mechanical Turk (AMT), and can dramatically reduce the cost and increase the throughput of biocuration efforts. However, given the size of the biomedical literature, even information extraction via paid microtask platforms is not scalable. With our web-based application Mark2Cure (http://mark2cure.org), we demonstrate that NER tasks also can be performed by volunteer citizen scientists with high accuracy. We apply metrics from the Zooniverse Matrices of Citizen Science Success and provide the results here to serve as a basis of comparison for other citizen science projects. Further, we discuss design considerations, issues, and the application of analytics for successfully moving a crowdsourcing workflow from a paid microtask platform to a citizen science platform. To our knowledge, this study is the first application of citizen science to a natural language processing task.


Promoting oral cancer examinations to medical primary care providers: a cluster randomized trial.
To compare the percentage of patients who had an oral cancer examination (OCE) by their primary care provider (PCP) in medical clinics participating in a web-based education with poster reminder intervention to that of patients in control clinics. To also determine the effects for PCPs in medical clinics participating in the web-based education with poster reminder intervention as compared with those in control clinics regarding: a) index of knowledge of oral cancer risk factors (RiskOC) and b) index of knowledge of oral cancer diagnostic procedures (DiagOC). Six medical clinics were recruited to participate in this study and randomly assigned to an intervention group or a control group. PCPs (physicians, physician assistants, and advanced practice registered nurses) took a pretest; 2 weeks later, they participated in the web-based educational program, including a posttest (intervention group) or took a posttest only (control group). In each clinic, 1 week following completion of the PCPs' posttests, 94 patients were recruited to complete a one-page survey. The intervention clinics were found to be a significant factor for the PCPs to perform patient OCEs, after controlling for significant covariates, that is, age, main reason for clinic visit, OCE for patient in the past year, clinic's mean DiagOC score, and clinic's mean RiskOC score. The intervention also resulted in the PCPs increasing their pretest to posttest RiskOC scores. The use of intervention has the potential to increase PCPs' short-term knowledge and to increase the frequency of PCPs' routine, nonsymptomatic opportunistic OCE on patients.


Resting State and Diffusion Neuroimaging Predictors of Clinical Improvements Following Constraint-Induced Movement Therapy in Children With Hemiplegic Cerebral Palsy.
The aim was to identify neuroimaging predictors of clinical improvements following constraint-induced movement therapy. Resting state functional magnetic resonance and diffusion tensor imaging data was acquired in 7 children with hemiplegic cerebral palsy. Clinical and magnetic resonance imaging (MRI) data were acquired at baseline and 1 month later following a 3-week constraint therapy regimen. A more negative baseline laterality index characterizing an atypical unilateral sensorimotor resting state network significantly correlated with an improvement in the Canadian Occupational Performance Measure score (r = -0.81, P = .03). A more unilateral network with decreased activity in the affected hemisphere was associated with greater improvements in clinical scores. Higher mean diffusivity in the posterior limb of the internal capsule of the affect tract correlated significantly with improvements in the Jebsen-Taylor score (r = -0.83, P = .02). Children with more compromised networks and tracts improved the most following constraint therapy.


The influence of smoking on plasma folate and lipoproteins in Alzheimer disease, mild cognitive impairment and depression.
The risk for Alzheimer's disease (AD) is associated with lifestyle factors, especially cigarette smoking. In this study we investigated the influence of smoking on the serum levels of folic acid, LDL and HDL in AD patients, patients with minimal cognitive impairment (MCI) and patients with major depression. We investigated a total of n = 374 patients in the diagnostic categories:, AD: n = 272, MCI: n = 60, Major depression: n = 42. We found significantly lower HDL levels in smokers and previous smokers in comparison to non-smokers, p<0,05. The LDL: HDL ratio in smokers was significant higher (+20%) compared to previous smokers and non-smokers, p < 0.05. The mean levels of folic acid were statistically significant (p<0.05) lower (-24%) in smokers compared to non-smokers. Patients with MCI and Alzheimer;s disease (and also major depression) who are "smokers" show serum levels of HDL and folic acid that are known to be strong risk factors for vascular damage and increased risk for vascular brain damage and impaired cognitive function. Therefore cessation of smoking, substitution with folate or statin therapy of smoking patients with MCI or AD might be beneficial to slow down further cognitive decline.


Mucoadhesive thin films for the simultaneous delivery of microbicide and anti-inflammatory drugs in the treatment of periodontal diseases.
There is an unmet clinical need for new products to address the high percentage of the populous who present with periodontal diseases. Drug dose retention at the point of application would facilitate sustained release and more efficacious treatments. The aim of this study was to evaluate mucoadhesive polymeric thin films for simultaneous in situ delivery chlorhexidine and anti-inflammatory and analgesic drugs. Mucoadhesive thin films were prepared using a polymer mixture containing chlorhexidine (25 mg) ± diclofenac sodium (10 and 50 mg), and lidocaine hydrochloride (10 mg) or betamethasone dipropionate (10 and 50 mg). The films were assessed for in vitro drug release and localised tissue delivery, followed by determination of modulated prostaglandin E2 (PGE2) levels in ex vivo tissue and cytotoxicity using a HaCaT keratinocyte cell line. Antibacterial activity of the chlorhexidine/diclofenac film was determined against planktonic and biofilm bacteria associated with periodontal disease and dental plaque. Chlorhexidine release was consistently low (up to 10% of initial loading) from all films, whereas the release of diclofenac, betamethasone and lidocaine exceeded 50% within 30 min. The 50 mg betamethasone film released up to 4-fold more than the 10 mg film. Statistically significant reduction of PGE2 was observed in ex vivo porcine gingival tissue for films containing chlorhexidine with or without diclofenac, and betamethasone. No cytotoxicity was observed for any film, apart from 50 mg betamethasone at 24 h. Films loaded with chlorhexidine and diclofenac were inhibitory against relevant test bacteria. Between 3 and 6 log10 reductions in bacterial cell recovery was observed after biofilm exposure to the chlorhexidine films irrespective of the presence of the anti-inflammatory or anaesthetic. This work demonstrated that thin film formulations have the potential to simultaneously counter key causative factors in periodontal diseases, namely associated bacteria biofilm and chronic local inflammation.


Defining whole-grain foods - does it change estimations of intakes and associations with CVD risk factors: an Australian and Swedish perspective.
Historically, there are inconsistencies in the calculation of whole-grain intake, particularly through use of highly variable whole-grain food definitions. The current study aimed to determine the impact of using a whole-grain food definition on whole-grain intake estimation in Australian and Swedish national cohorts and investigate impacts on apparent associations with CVD risk factors. This utilised the Australian National Nutrition and Physical Activity Survey 2011-2012, the Swedish Riksmaten adults 2010-2011 and relevant food composition databases. Whole-grain intakes and associations with CVD risk factors were determined based on consumption of foods complying with the Healthgrain definition (≥30 % whole grain (dry weight), more whole than refined grain and meeting accepted standards for 'healthy foods' based on local regulations) and compared with absolute whole-grain intake. Compliance of whole-grain containing foods with the Healthgrain definition was low in both Sweden (twenty-nine of 155 foods) and Australia (214 of 609 foods). Significant mean differences of up to 24·6 g/10 MJ per d of whole-grain intake were highlighted using Swedish data. Despite these large differences, application of a whole-grain food definition altered very few associations with CVD risk factors, specifically, changes with body weight and blood glucose associations in Australian adults where a whole-grain food definition was applied, and some anthropometric measures in Swedish data where a high percentage of whole-grain content was included. Use of whole-grain food definitions appears to have limited impact on measuring whole-grain health benefits but may have greater relevance in public health messaging.


Speed and direction of locally-paired dot patterns.
Phenomenal transparency in random-dot kinematograms is abolished when two motion directions are 'locally-balanced' by pairing limited-lifetime dots at each location [Qian, Andersen and Adelson (1994). Journal of Neuroscience, 14, 7357-7366]. Qian et al. also report that locally-paired stimuli appear as directionless flicker when the paired dots differ in their directions by 90 degrees or more. They attribute this to local inhibition between motion detectors more than 45 degrees apart. We investigated perceived motion in such displays, by requiring subjects to make direction and speed judgements with locally-paired stimuli containing two directions 60, 90 or 120 degrees apart. Subjects perceived coherent motion in these displays and made reliable direction judgements, indicating that the two motions are combined rather than interfering destructively. Our results show that the judged motion of locally-paired stimuli is in the vector-average direction of the two components. This vector-averaging rule also applies when the two sets of component dots differ in their velocity. Similarly, speed judgements comply with a vector-averaging rule for a range of speeds as well as for mixed-speed stimuli. These results suggest that the abolition of transparency does not necessarily imply abolition of a global motion percept. The local interaction abolishing transparency is not exclusively inhibitory, at least for directions up to 120 degrees apart, but generates a vector combination of the superimposed motions.


Use of an integrated Atlas of Mental Health Care for evidence informed policy in Catalonia (Spain).
This paper aims to present the Integrated Atlas of Mental Health of Catalonia (2010) focusing on: (a) the importance of using a taxonomy-based coding and standard system of data collection when assessing health services; and (b) its relevance as a tool for evidence-informed policy. This study maps all the care-related services for people with mental disorders available in Catalonia in 2010, using the 'Description and Evaluation of Services and Directories in Europe for long-term care' (DESDE-LTC). The unit of analysis is the Basic Stable Input of Care (BSIC), which is the minimal organisation unit composed by a set of inputs with temporal stability. We presented data on: (a) availability of BSICs and their capacity; (b) the adequacy of the provision of care, taking into account availability and accessibility; (c) the evolution of BSCIs from 2002 to 2010; and (d) the perceived relevance of Atlas of Mental Health as a tool for evidence-informed policy. We identified a total of 639 BSICs. A lack of Health services was detected in highly rural areas, although there was moderate availability of Social Services. Overall, more than 80% of the small mental health areas in Catalonia had an adequate core mental health service. Since 2002 the availability of mental health services has increased. Decision makers found the Atlas a useful and relevant tool for evidence informed policy. Policy makers can use Atlases to detect gaps and inequities in the provision of care for people with mental health needs.


Short-term predictors of maintenance of normotension after withdrawal of antihypertensive drugs in the second Australian National Blood Pressure Study (ANBP2).
Antihypertensive drug therapy is considered lifelong but in the family practice environment drug cessation may provide an opportunity to attempt nonpharmacologic strategies for blood pressure (BP) control with a clear outcome, maintaining drug-free status. The identification of simple predictors would assist the family physician to select who may or may not have their medication ceased. To monitor a drug cessation program in currently treated hypertensive patients in Australian family practice, 25,826 patients aged 65 to 84 years currently receiving antihypertensive medication, were offered drug withdrawal as part of the run-in phase of a large clinical trial. Outcomes investigated were the proportion of patients completing drug withdrawal and maintaining short-term BP control and factors that predicted these patients. A total of 18,993 patients did not enter the withdrawal program; 6291 (92% of those who entered) completed drug withdrawal. In comparison to patients who did not complete drug withdrawal, they were younger and more likely male. A total of 1,228 (18% of those who entered) ceased medication and maintained adequate BP control for 0 to 76 weeks (median, 4 weeks). Cox regression analysis identified lower on therapy systolic and diastolic BP, younger age, type of agent, and monotherapy as predictors of successful drug withdrawal and maintenance of BP control. Cessation of antihypertensive drug therapy is possible in a substantial proportion (18%) of patients attending family practice who are willing to do so and is most successful in those who are younger with BP controlled on monotherapy. Where this strategy is initiated, use of such predictors, effective behavioral change, and systematic follow-up is recommended.


First microbiota assessments of children's paddling pool waters evaluated using 16S rRNA gene-based metagenome analysis.
Insufficient chloric sterilization of children's paddling pool waters increases the risk of diarrheal illness. Therefore, we investigated the microbiota changes after children use pools. First, we applied 16S rRNA gene-based metagenome analysis to understand the dynamics of microbiota in pool water, especially with respect to the bio-contamination by potential pathogens. Proteobacteria were major taxa detected in every pool water sample after children spent time in the pool. In more detail, Gammaproteobacteria comprised the dominant class, which was followed by Betaproteobacteria. Five phyla, Bacteroidetes, Firmicutes, Actinobacteria and Deinococcus-Thermus phyla were minor groups. The pool water microbiota are likely to be a consortium of intestinal and skin microbiota from humans. Interestingly, the ratio of Gammaproteobacteria and Betaproteobacteria differed according to the age of the children who used the pool, which means the pool water was additionally contaminated by soil microbiota as a result of the children's behavior. Furthermore, potential pathogens, such as Campylobacter spp., Comamonas testosteroni and Burkholderia pseudomallei, were also found. Considering the standard plate counts, the abundances of these human pathogens are unlikely to be a sufficiently infectious dose. We suggest the importance of sanitary measures in paddling pool waters to reduce bio-contamination from both humans and the environment.


Does Rheumatoid Cachexia Predispose Patients with Rheumatoid Arthritis to Osteoporosis and Vertebral Fractures?
To assess the prevalence and risk factors of rheumatoid cachexia (RC) and evaluate its relationship with osteoporosis and vertebral fractures (VF) in patients with rheumatoid arthritis (RA). We enrolled into a cross-sectional study 178 consecutive patients with RA (82.6% women) with a mean age of 54.1 ± 11.5 years (25-82) and who fulfilled the American College of Rheumatology criteria for the classification of RA. Body composition, lateral VF assessment images, and scans of the lumbar spine and proximal femur were obtained using dual-energy x-ray absorptiometry. RC was defined by a fat-free mass index below the 10th percentile and a fat mass index above the 25th percentile compared with a reference population. VF were defined using Genant semiquantitative approach. RC was observed in 96 patients (53.9%) and osteoporosis in 52 patients (29.2%). Comparison between women with and without RC showed that women with RC had a longer disease duration, higher disease activity variables, higher steroid cumulative dose, and higher proportion of patients with erosive arthritis. Women with RC had lower total hip bone mineral density (BMD) and T score than women without RC, while comparison in men found only body mass index to be significantly lower in men with RC. Regression logistic analysis showed an independent and significant association between RC and age and disease activity in women. Our study showed that half of the patients with RA may have RC, a condition that was significantly associated with disease activity and low hip BMD, but not with VF.


Stronger synaptic connectivity as a mechanism behind development of working memory-related brain activity during childhood.
The cellular maturational processes behind cognitive development during childhood, including the development of working memory capacity, are still unknown. By using the most standard computational model of visuospatial working memory, we investigated the consequences of cellular maturational processes, including myelination, synaptic strengthening, and synaptic pruning, on working memory-related brain activity and performance. We implemented five structural developmental changes occurring as a result of the cellular maturational processes in the biophysically based computational network model. The developmental changes in memory activity predicted from the simulations of the model were then compared to brain activity measured with functional magnetic resonance imaging in children and adults. We found that networks with stronger fronto-parietal synaptic connectivity between cells coding for similar stimuli, but not those with faster conduction, stronger connectivity within a region, or increased coding specificity, predict measured developmental increases in both working memory-related brain activity and in correlations of activity between regions. Stronger fronto-parietal synaptic connectivity between cells coding for similar stimuli was thus the only developmental process that accounted for the observed changes in brain activity associated with development of working memory during childhood.


Dark Classics in Chemical Neuroscience: 3,4-Methylenedioxymethamphetamine.
Better known as "ecstasy", 3,4-methylenedioxymethamphetamine (MDMA) is a small molecule that has played a prominent role in defining the ethos of today's teenagers and young adults, much like lysergic acid diethylamide (LSD) did in the 1960s. Though MDMA possesses structural similarities to compounds like amphetamine and mescaline, it produces subjective effects that are unlike any of the classical psychostimulants or hallucinogens and is one of the few compounds capable of reliably producing prosocial behavioral states. As a result, MDMA has captured the attention of recreational users, the media, artists, psychiatrists, and neuropharmacologists alike. Here, we detail the synthesis of MDMA as well as its pharmacology, metabolism, adverse effects, and potential use in medicine. Finally, we discuss its history and why it is perhaps the most important compound for the future of psychedelic science-having the potential to either facilitate new psychedelic research initiatives, or to usher in a second Dark Age for the field.


Genome-wide association analysis identifies new lung cancer susceptibility loci in never-smoking women in Asia.
To identify common genetic variants that contribute to lung cancer susceptibility, we conducted a multistage genome-wide association study of lung cancer in Asian women who never smoked. We scanned 5,510 never-smoking female lung cancer cases and 4,544 controls drawn from 14 studies from mainland China, South Korea, Japan, Singapore, Taiwan and Hong Kong. We genotyped the most promising variants (associated at P < 5 × 10(-6)) in an additional 1,099 cases and 2,913 controls. We identified three new susceptibility loci at 10q25.2 (rs7086803, P = 3.54 × 10(-18)), 6q22.2 (rs9387478, P = 4.14 × 10(-10)) and 6p21.32 (rs2395185, P = 9.51 × 10(-9)). We also confirmed associations reported for loci at 5p15.33 and 3q28 and a recently reported finding at 17q24.3. We observed no evidence of association for lung cancer at 15q25 in never-smoking women in Asia, providing strong evidence that this locus is not associated with lung cancer independent of smoking.


Oocyte activation and phospholipase C zeta (PLCζ): diagnostic and therapeutic implications for assisted reproductive technology.
Infertility affects one in seven couples globally and has recently been classified as a disease by the World Health Organisation (WHO). While in-vitro fertilisation (IVF) offers effective treatment for many infertile couples, cases exhibiting severe male infertility (19-57%) often remain difficult, if not impossible to treat. In such cases, intracytoplasmic sperm injection (ICSI), a technique in which a single sperm is microinjected into the oocyte, is implemented. However, 1-5% of ICSI cycles still fail to fertilise, affecting over 1000 couples per year in the UK alone. Pregnancy and delivery rates for IVF and ICSI rarely exceed 30% and 23% respectively. It is therefore imperative that Assisted Reproductive Technology (ART) protocols are constantly modified by associated research programmes, in order to provide patients with the best chances of conception. Prior to fertilisation, mature oocytes are arrested in the metaphase stage of the second meiotic division (MII), which must be alleviated to allow the cell cycle, and subsequent embryogenesis, to proceed. Alleviation occurs through a series of concurrent events, collectively termed 'oocyte activation'. In mammals, oocytes are activated by a series of intracellular calcium (Ca2+) oscillations following gamete fusion. Recent evidence implicates a sperm-specific phospholipase C, PLCzeta (PLCζ), introduced into the oocyte following membrane fusion as the factor responsible. This review summarises our current understanding of oocyte activation failure in human males, and describes recent advances in our knowledge linking certain cases of male infertility with defects in PLCζ expression and activity. Systematic literature searches were performed using PubMed and the ISI-Web of Knowledge. Databases compiled by the United Nations and World Health Organisation databases (UNWHO), and the Human Fertilization and Embryology Authority (HFEA) were also scrutinised. It is clear that PLCζ plays a fundamental role in the activation of mammalian oocytes, and that genetic, molecular, or biochemical perturbation of this key enzyme is strongly linked to human infertility where oocyte activation is deficient. Consequently, there is significant scope for our understanding of PLCζ to be translated to the ART clinic, both as a novel therapeutic agent with which to rescue oocyte activation deficiency (OAD), or as a prognostic/diagnostic biomarker of oocyte activation ability in target sperm samples.


Characterization of antibody V segment diversity in the Tasmanian devil (Sarcophilus harrisii).
The Tasmanian devil (Sarcophilus harrisii) immune system has recently been under scrutiny because of the emergence of a contagious cancer, which has decimated devil numbers. Here we provide a comprehensive description of the Tasmanian devil immunoglobulin variable regions. We show that heavy chain variable (VH) and light chain variable (VL) repertoires are similar to those described in other marsupial taxa: VL diversity is high, but VH diversity is restricted and belongs only to clan III. As in other mammals, one VH and one Vλ germline family and multiple incomplete Vκ germline sequences were identified in the genome. High Vκ variation was observed in transcripts and we predict that it may have arisen by gene conversion and/or somatic mutations, as it does not appear to have originated from germline variation. Phylogenetic analyses revealed that devil VL gene segments are highly complex and ancient, with some lineages predating the separation of marsupials and eutherians. These results indicate that although the evolutionary history of immune genes lead to the expansions and contractions of immune gene families between different mammalian lineages, some of the ancestral immune gene variants are still maintained in extant species. A high degree of similarity was found between devil and other marsupial VH segments, demonstrating that they originated from a common clade of closely related sequences. The VL families had a higher variation than VH both between and within species. We suggest that, similar to other studied marsupial species, the complex VL segment repertoire compensates for the limited VH diversity in Tasmanian devils.


Radiation response of mesenchymal stem cells derived from bone marrow and human pluripotent stem cells.
Mesenchymal stem cells (MSCs) isolated from human pluripotent stem cells are comparable with bone marrow-derived MSCs in their function and immunophenotype. The purpose of this exploratory study was comparative evaluation of the radiation responses of mesenchymal stem cells derived from bone marrow- (BMMSCs) and from human embryonic stem cells (hESMSCs). BMMSCs and hESMSCs were irradiated at 0 Gy (control) to 16 Gy using a linear accelerator commonly used for cancer treatment. Cells were harvested immediately after irradiation, and at 1 and 5 days after irradiation. Cell cycle analysis, colony forming ability (CFU-F), differentiation ability, and expression of osteogenic-specific runt-related transcription factor 2 (RUNX2), adipogenic peroxisome proliferator-activated receptor gamma (PPARγ), oxidative stress-specific dismutase-1 (SOD1) and Glutathione peroxidase (GPX1) were analyzed. Irradiation arrested cell cycle progression in BMMSCs and hESMSCs. Colony formation ability of irradiated MSCs decreased in a dose-dependent manner. Irradiated hESMSCs showed higher adipogenic differentiation compared with BMMSCs, together with an increase in the adipogenic PPARγ expression. PPARγ expression was upregulated as early as 4 h after irradiation, along with the expression of SOD1. More than 70% downregulation was found in Wnt3A, Wnt4, Wnt 7A, Wnt10A and Wnt11 in BMMSCs, but not in hESMSCs. hESMSCs are highly proliferative but radiosensitive compared with BMMSCs. Increased PPARγ expression relative to RUNX2 and downregulation of Wnt ligands in irradiated MSCs suggest Wnt mediated the fate determination of irradiated MSCs.


Renal cell carcinoma in the Canadian Indigenous population.
Diagnosis and treatment of renal cell carcinoma (rcc) might be different in Indigenous Canadians than in non-Indigenous Canadians. In this cohort study, we compared rcc presentation and treatments in Indigenous and non-Indigenous Canadians. Patients registered in the Canadian Kidney Cancer Information System treated at 16 institutions between 2011 and 2018 were included. Baseline patient, tumour, and treatment characteristics were compared between Indigenous and non-Indigenous Canadians. The primary objective was to determine if differences in rcc stage at diagnosis were evident between the groups. The secondary objective was to determine if treatments and outcomes were different between the groups. During the study period, 105 of the 4529 registered patients self-identified as Indigenous. Those patients were significantly younger at the time of clinical diagnosis (57.9 ± 11.3 years vs. 62.0 ± 12.1 years, p = 0.0006) and had a family history prevalence of rcc that was double the prevalence in the non-Indigenous patients (14% vs. 7%, p = 0.004). Clinical stage at diagnosis was similar in the two groups (p = 0.61). The disease was metastatic at presentation in 11 Indigenous Canadians (10%) and in 355 non-Indigenous Canadians (8%). Comorbid conditions that could affect the management of rcc-such as obesity, renal disease, diabetes mellitus, and smoking-were more common in Indigenous Canadians (p < 0.05). Indigenous Canadians experienced a lower rate of active surveillance (p = 0.01). Treatments and median time to treatments were similar in the two groups. Compared with their non-Indigenous counterparts, Indigenous Canadian patients with rcc are diagnosed at an earlier age and at a similar clinical stage. Despite higher baseline comorbid conditions, clinical outcomes are not worse for Indigenous Canadians than for non-Indigenous Canadians.


A retrospective study on the unseen epidemic of road traffic injuries and deaths due to accidents in Mwanza City - Tanzania.
Sixty percent of the global deaths and injuries occur in the developing world and mostly are due to Road traffic accidents (RTAs. looking at the etiological related factors which include, carelessness of the driver, condition of the vehicle or motorcycle, poor condition of roads, risky behavior of the driver, most of these factors can be prevented to some extent. This study therefore, determined the pattern of cases and deaths due to traffic road accidents in Mwanza City Tanzania. In this retrospective study, records, registers and case notes In the surgical ward and causality, medical records and central police station from 2008 to 2011 were used. The study focused on the two referral hospitals (Sekouture regional hospital and Bugando Medical Center). There were 3450 cases due to accidents reported at both centers (Sekouture regional hospital and Bugando Medical Center of which 3224 (93.4%) had complete information for analysis.2225 (69%) were male and 999 (31%) were female, and the most affected group were male. Among the RTAs2809 cases (87%) were due to motor cycle accidents which were the leading cause of RTAs with case fatality rate of 5% while motor vehicle has case fatality rate of 24% which is 5 times that of motor cycle. Among all RTAs the leading cause of injuries is Motor cycle traffic accidents followed by motor vehicle. RTAs are on increase particularly the motor cycle traffic accidents and has claimed a good number of innocent people's lives however most of them are preventable, therefore driving course to be introduced to motor cycle drivers with emphasize on the road posters signal, rules and regular checkup of their motor cycles especially commercial motor cycle.


Isolation, purification and biological activity of major docetaxel metabolites from human feces.
We have developed a procedure suited for the isolation of metabolites of docetaxel (Taxotere) from human feces. The compounds were extracted from the feces with diethyl ether and further purified by (semipreparative) HPLC. Four metabolic products were obtained in submilligram quantities. Analytical HPLC with photodiode array detection showed that the purity of each compound was higher than 98%. There structures have been characterized by UV absorption and FAB/MS. All four compounds were oxidation products of the tert-butyl group attached to the C13-side chain, and corresponded to structures identified previously. The purified products were used for evaluating their cytotoxic activities against a human ovarian cancer (A2780) and a rat colon cancer (CC531) cell line, and their myelosuppressive effects in a hematopoietic progenitor toxicity assay. Although distinctions in biological activities between the compounds were evident, all metabolites showed a marked reduction in both cytotoxic and myelotoxic properties.


The hypothesis of an effective safe and novel radioprotective agent: hydrogen-rich solution.
Most ionizing radiation-induced damage is caused by radical oxygen species (ROS). Some radioprotectors, such as amifostine, exert radioprotective effects by scavenging radical oxygen species. Recent studies show that hydrogen (H) has antioxidant activities that protect the brain and intestine against ischaemia-reperfusion injury and stroke by selectively reducing hydroxyl and peroxynitrite radicals. However it is seldom regarded as a radioprotective agent. In like manner we hypothesize that hydrogen may be an effective, specific and novel radioprotective agent. But H2 is explosive, while hydrogen-rich solution (solution such as physiological saline saturated with molecular hydrogen) is safer.


[Validation of noise reduction in iterative reconstruction: a simulation phantom study].
In the iterative reconstruction method, image noise tends to increase in proportion to falling available photon count and increasing update number. Image filtering is an important factor in single photon emission computed tomography (SPECT) image reconstruction, but it is frequently treated in a subjective way. The aim of this study was to evaluate the effects of pre-reconstruction filtering and post-reconstruction filtering on the iterative reconstruction process. Using simulation phantoms, projection data were reconstructed using ordered subsets expectation maximization (OSEM) with or without compensation for resolution recovery. Pre- and post-reconstruction filtering was performed using a Butterworth filter (BW) (range: 0.3-1.3 cycles/cm) and a Gaussian filter (GA) (range: 0.3-1.3 mm) with various parameters. We evaluated the variances of full width at half maximum (FWHM), coefficients of variation (CV), image contrast and normalized mean squared error (NMSE) values. The FWHM values for pre-reconstruction filtering tended to be lower than those observed for post-reconstruction filtering. These values were 5.1 mm (pre-reconstruction) and 6.7 mm (post-reconstruction). The CV on pre- and post-reconstruction filtering was 7.5% and 11.6%. Pre-reconstruction filtering reduced image noise more effectively than post-reconstruction filtering. The contrast for pre-reconstruction filtering was similar to that observed after post-reconstruction filtering. However, contrast after filtering with a GA slowly decreased as compared to the BW. NMSE values obtained by pre-reconstruction filtering tended to be lower than those observed for post-reconstruction filtering. Pre-reconstruction filtering provided SPECT image quality comparable to that from post-reconstruction filtering, especially when using the BW. Our results suggest that pre-reconstruction filtering is a beneficial method when applied to the iterative reconstruction method with or without compensation for resolution recovery.


Synthesis of the Ca2+-mobilizing messengers NAADP and cADPR by intracellular CD38 enzyme in the mouse heart: Role in β-adrenoceptor signaling.
Nicotinic acid adenine dinucleotide phosphate (NAADP) and cyclic ADP-ribose (cADPR) are Ca2+-mobilizing messengers important for modulating cardiac excitation-contraction coupling and pathophysiology. CD38, which belongs to the ADP-ribosyl cyclase family, catalyzes synthesis of both NAADP and cADPR in vitro However, it remains unclear whether this is the main enzyme for their production under physiological conditions. Here we show that membrane fractions from WT but not CD38-/- mouse hearts supported NAADP and cADPR synthesis. Membrane permeabilization of cardiac myocytes with saponin and/or Triton X-100 increased NAADP synthesis, indicating that intracellular CD38 contributes to NAADP production. The permeabilization also permitted immunostaining of CD38, with a striated pattern in WT myocytes, whereas CD38-/- myocytes and nonpermeabilized WT myocytes showed little or no staining, without striation. A component of β-adrenoreceptor signaling in the heart involves NAADP and lysosomes. Accordingly, in the presence of isoproterenol, Ca2+ transients and contraction amplitudes were smaller in CD38-/- myocytes than in the WT. In addition, suppressing lysosomal function with bafilomycin A1 reduced the isoproterenol-induced increase in Ca2+ transients in cardiac myocytes from WT but not CD38-/- mice. Whole hearts isolated from CD38-/- mice and exposed to isoproterenol showed reduced arrhythmias. SAN4825, an ADP-ribosyl cyclase inhibitor that reduces cADPR and NAADP synthesis in mouse membrane fractions, was shown to bind to CD38 in docking simulations and reduced the isoproterenol-induced arrhythmias in WT hearts. These observations support generation of NAADP and cADPR by intracellular CD38, which contributes to effects of β-adrenoreceptor stimulation to increase both Ca2+ transients and the tendency to disturb heart rhythm.


Oxidation of aromatic alcohols in irradiated aqueous suspensions of commercial and home-prepared rutile TiO(2): a selectivity study.
The photocatalytic oxidation of benzyl alcohol (BA) and 4-methoxybenzyl alcohol (MBA) has been performed in pure water by using commercial TiO(2) samples (Sigma-Aldrich, Merck, Degussa P25) and rutile TiO(2) prepared from TiCl(4) at low temperature. Particular attention has been devoted to the identification of the produced aromatic compounds along with the formed CO(2). Oxidation products such as the corresponding aromatic aldehyde and acid, as well as mono- and dihydroxylated aldehydes have been detected. The home-prepared rutile sample showed a marked selectivity towards the formation of the aromatic aldehyde (38 and 60 % for BA and MBA, respectively), resulting in a three- to sevenfold improvement relative to commercial samples, with the only byproduct being CO(2). This catalyst was found to be the most selective in the formation of aldehyde in water. By using the commercial or the calcined home-prepared samples, many hydroxylated aromatic compounds were detected besides the aldehyde and the acid. This finding points to a higher selectivity performance of the home-prepared rutile relative to the commercial TiO(2) samples. Some of the home-prepared samples were also dialysed to check the influence of the presence of Cl(-) species on catalyst reactivity and selectivity. We have attempted to explain the different reaction rate and selectivity observed for MBA and BA.


[A compound heterozygous mutation in CYP17A1 gene in a female subject with partial 17α-hydroxylase/17, 20 lyase deficiency].
To explore the clinical and molecular genetic characteristics of a Chinese female patient with partial 17α-hydroxylase/17, 20 lyase deficiency (17OHD), a rare type of congenital adrenal hyperplasia. Her clinical features and laboratory data were collected. Genomic DNA was extracted from leukocytes of peripheral blood of her and her mother. All eight exons of CYP17A1 gene, including flanking regions of introns, were amplified by PCR. The mutations of CYP17A1 gene were identified by direct sequencing or cloning and sequencing the amplified DNA fragments. The patient presented with hypertension, hypokalemia and irregular menstruation. DNA sequencing results demonstrated a compound heterozygous mutation in CYP17A1 gene. One allele of her had the deletion of phenylalanine (TTC) at either codon 53 or 54 and the other allele contained a base transversion at codon 329 (TAC/AA) and leading to a missense mutation of tyrosine to lysine and the open reading frame shift following this codon to produce a truncated enzyme with 417 amino acids and without activity site. Her mother was a heterozygous carrier of the latter allele. The partial 17OHD in this patient is caused by a compound heterozygous mutation in CYP17A1 gene.


Developments in transgenic fish in the People's Republic of China.
In the People's Republic of China, genetically modified (GM) fish are being developed primarily to produce desirable alterations to growth rates or feed-conversion efficiency. Up to the present, no transgenic fish have been commercially approved for human consumption. This review introduces advances in the People's Republic of China in transgenic fish studies, biosafety studies of fast-growth GM fish, and the regulation of GM fish.


Harmonization of HTA--based reimbursement and regulatory approval activities: a qualitative study.
Regulatory approval and reimbursement are necessary to bring health technologies to market. As both are distinct processes, the lack of interaction between regulators and payers has been criticized for resulting in health system inefficiencies. This study describes the experiences of health system stakeholders on the development and implementation of initiatives to better harmonize HTA-reimbursement and regulatory activities, as well as Canadian perceptions on such initiatives. Thematic analysis of semi-structured interviews was conducted. Participants represented those whom have published or worked actively in this area or who held positions in HTA and/or reimbursement agencies that gave them insight into the Canadian healthcare system. Interviews were recorded, transcribed and analyzed to identify emerging themes and relationships. Fourteen individuals from twelve organizations participated, including representatives from Canada, US, UK and Netherlands. Harmonization was identified as either a means to: (i) develop economies of scale in the generation of clinical data; and/or (ii) align the lifecycle of a health technology. For such initiatives to be successfully implemented, the following key issues emerged: (i) fostering healthy relationships; (ii) promoting well-intentions; (iii) defining governance and leadership clearly; and (iv) securing organizational infrastructure. Attempts to harmonize reimbursement and regulatory activities is in its infancy; although, much can be learned from current Canadian and international experiences. Within Canada there is much interest, although concerns have been raised on the extent to which harmonization is possible. Successful policy implementation would require inter-agency collaborations, with particular attention towards capacity building and strong leadership.


Comparison of air sac volume, lung volume, and lung densities determined by use of computed tomography in conscious and anesthetized Humboldt penguins (Spheniscus humboldti) positioned in ventral, dorsal, and right lateral recumbency.
To determine the effects of recumbency on air sac volume, lung volume, and lung densities in CT images of healthy, conscious and anesthetized spontaneously breathing Humboldt penguins (Spheniscus humboldti). 25 adult (13 male and 12 female) Humboldt penguins. CT images of conscious penguins in ventral recumbency and anesthetized penguins in dorsal, ventral, and right lateral recumbency were obtained. Air sac volume, lung volume, and lung densities in CT images were calculated. A paired samples t test was used to determine whether right and left lung densities differed among recumbencies. Repeated-measures ANOVA (controlled for sex and order of recumbencies during CT) was used to determine whether air sac or lung volumes differed among recumbencies. Recumbency had a significant effect on air sac volume but not lung volume. Air sac volume was largest in conscious penguins in ventral recumbency (mean ± SD, 347.2 ± 103.1 cm(3)) and lowest in anesthetized penguins in dorsal recumbency (median, 202.0 cm(3); 10th to 90th percentile, 129.2 to 280.3 cm(3)). Lung densities were highest in anesthetized penguins in dorsal recumbency (right lung median, 0.522 g/cm(3); left lung median, 0.511 g/cm(3)) and lowest in anesthetized penguins in ventral recumbency (right lung median, 0.488 g/cm(3); left lung median, 0.482 g/cm(3)). Results indicated that anesthetized Humboldt penguins had the lowest air sac volume and highest lung densities in dorsal recumbency. Therefore, this recumbency may not be recommended. Minimal changes in lung volume were detected among recumbencies or between conscious and anesthetized penguins.


Reflecting upon feelings: an fMRI study of neural systems supporting the attribution of emotion to self and other.
Understanding one's own and other individual's emotional states is essential for maintaining emotional equilibrium and strong social bonds. Although the neural substrates supporting ref lection upon one's own feelings have been investigated, no studies have directly examined attributions about the internal emotional states of others to determine whether common or distinct neural systems support these abilities. The present study sought to directly compare brain regions involved in judging one's own, as compared to another individual's, emotional state. Thirteen participants viewed mixed valence blocks of photos drawn from the International Affective Picture System while whole-brain fMRI data were collected. Preblock cues instructed participants to evaluate either their emotional response to each photo, the emotional state of the central figure in each photo, or (in a baseline condition) whether the photo was taken indoors or outdoors. Contrasts indicated (1) that both self and other judgments activated the medial prefrontal cortex (MPFC), the superior temporal gyrus, and the posterior cingulate/precuneus, (2) that self judgments selectively activated subregions of the MPFC and the left temporal cortex, whereas (3) other judgments selectively activated the left lateral prefrontal cortex (including Broca's area) and the medial occipital cortex. These results suggest (1) that self and other evaluation of emotion rely on a network of common mechanisms centered on the MPFC, which has been hypothesized to support mental state attributions in general, and (2) that medial and lateral PFC regions selectively recruited by self or other judgments may be involved in attention to, and elaboration of, internally as opposed to externally generated information.


Förster resonance energy transfer to impart signal-on and -off capabilities in a single microRNA biosensor.
Many studies have found that over- or under-expression of biomolecules called microRNAs (miRNA) regulates several diseases. Biosensors are in need to visually identify the relative expression level of miRNA to determine the direction these miRNA change in cells and tissues. Our established reporter+probe miRNA biosensor design requires that miRNA outcompete and displace the reporter from the probe. Once displaced, the reporter folds into a hairpin structure to force together a pair of Förster Resonance Energy Transfer (FRET) dyes. The donor and acceptor signal changes can be used to indicate the over-/under-expression of miRNA. The bright signal from the donor will indicate miRNA under-expression; the bright acceptor signal will indicate miRNA over-expression. Since close proximity of the dyes to each other and nucleic acids often quench fluorescence, polyethylene glycol spacers were added in-between the dyes and nucleic acids. We compared reporter designs with and without spacers to investigate the effects on the following analytical metrics: (1) extent of signal change, (2) limits of detection and quantitation, and (3) sensitivity. Systematic errors and amount of reporter+probe biosensor formed were evaluated for one of the biosensors. Cy3|Cy5 and 6-carboxyfluorescein (6-FAM)|ATTO 633 dye pairs on reporters containing spacers showed an increase in the acceptor signal change by ∼190 and ∼484%, respectively, compared to no spacers. Transduction mechanisms that enhance and quench the signal both showed LODs that ranged from 3-17 nanomolar (nM) with 100 nM of the biosensor.


Successful use of steroids and ureteric stents in 24 patients with idiopathic retroperitoneal fibrosis: a retrospective study.
Retroperitoneal fibrosis (RPF) is a chronic inflammatory disorder causing obstructive nephropathy and renal failure. We reviewed our management of this condition. All patients with RPF treated at a single center over a 15-year period were identified. A full review of notes and computer records was undertaken. Data was available on 27 patients, 3 of which were excluded from later analysis. Diagnosis was based on clinical history and cross-sectional imaging. Retroperitoneal biopsy was undertaken in 3 patients. 96% had significant renal impairment at presentation with a mean serum creatinine of 688 micromol/l. 46% required emergency hemodialysis. All patients were treated with a combination of ureteric stents and/or steroids with an excellent clinical response. The mean best creatinine reached by the cohort was 136 micromol/l, and renal function remained stable in the long term. No patients required chronic dialysis. Ureteric stents were removed within 12 months and low-dose steroids were continued for a mean of 34 months. Recurrent disease was observed in 25% of patients, who all responded well to further steroid therapy. Mean duration of follow-up was 76 months. RPF is very effectively treated by a combination of ureteric stents and steroids, with excellent long-term results using this approach. Continued follow-up is advised because of the possibility of recurrent disease.


Biodiversity hotspots for conservation priorities.
Conservationists are far from able to assist all species under threat, if only for lack of funding. This places a premium on priorities: how can we support the most species at the least cost? One way is to identify 'biodiversity hotspots' where exceptional concentrations of endemic species are undergoing exceptional loss of habitat. As many as 44% of all species of vascular plants and 35% of all species in four vertebrate groups are confined to 25 hotspots comprising only 1.4% of the land surface of the Earth. This opens the way for a 'silver bullet' strategy on the part of conservation planners, focusing on these hotspots in proportion to their share of the world's species at risk.


First Report of Tomato Pith Necrosis Caused by Pseudomonas mediterranea in the United States and P. corrugata in Ohio.
Tomato (Solanum lycopersicum, cvs. Mountain Fresh, Big Dena, and Trust) plants with symptoms of pith necrosis were received from six commercial high tunnels in Ohio during May and July 2012. Disease incidence ranged from 1 to 5%. Symptoms included wilting of shoots, dry, dark brown coalescent lesions on stems, brown discolored pith with a ladder-like appearance, and in some cases, adventitious root formation. Bacterial streaming was observed microscopically from necrotic stem tissue. Bacteria were isolated from surface-sterilized diseased stem tissue by plating 10-fold serial dilutions onto yeast dextrose carbonate (YDC) and Pseudomonas F (PF) agar media. The majority of the colonies recovered were similar in morphology on YDC: round and mucoid, with a greenish center that later became dry and winkled with a curly margin, and producing a yellow-green diffusible pigment. Colonies were creamy, yellow-brown in color and non-florescent on PF medium. Nine isolates from six plant samples were purified. All isolates were gram-negative, levan negative, oxidase positive, and potato rot negative. Three isolates were positive and six were negative for arginine dihydrolase activity. None induced a hypersensitive reaction in tobacco. All isolates grew at 37°C. The isolates were further identified by PCR assays using species-specific primers PC5/1-PC5/2 for Pseudomonas corrugata and PC1/1-PC1/2 for P. mediterranea (1,2). DNA of a reference P. mediterranea strain from Turkey was used as a positive control. A 600-bp band was amplified using P. mediterranea primers from the six arginine dihydrolase negative isolates recovered from four of six samples. An 1,100-bp band was amplified from the three arginine dihydrolase positive isolates from two other samples using P. corrugata primers. The 600-bp PCR products amplified from the P. mediterranea reference strain and isolate SM664-12 were purified and sequenced. The DNA sequence of SM664-12 was 99% aligned with that of the reference strain from Turkey and a BLAST search in NCBI indicated only one match with P. mediterranea strain G-229-21 (Accession No. EU117098.1), with an E-value 1e-145 and 84% identity. P. mediterranea (SM664-12) and P. corrugata (SM658-12) were each inoculated onto four 4-week-old tomato plants (cv. Mountain Fresh) by injecting a 50 μl bacterial suspension (108 CFU/ml) into the stem at the axil of the first true leaf (2). Negative control plants were injected with sterile water. Plants were kept in a mist chamber for 72 h at 25°C, then moved into a growth chamber maintained at 25/20°C day/night, 12-h light/dark, and 80% relative humidity. Plants exhibited dark brown lesions at the inoculation site after 4 weeks and brown discoloration of the pith developed, whereas no lesions were observed in control plants. The reisolated bacteria were tested by PCR and identified as P. corrugata and P. mediterranea. Therefore, we have confirmed that tomato pith necrosis in Ohio involves at least two bacteria, P. corrugata and P. mediterranea. Although tomato pith necrosis has been observed in Ohio since the 1990s, to our knowledge, this is the first confirmation of a causal agent as P. corrugata in Ohio and the first report of P. mediterranea causing tomato pith necrosis in the United States. References: (1) V. Catara et al. Eur. J. Plant Pathol. 106:753, 2000. (2) V. Catara et al. Int. J. Syst. Evol. Microbiol. 52:1749, 2002.


Evaluation of Veterinary Students' Communication Skills with a Service Dog Handler in a Simulated Client Scenario.
Effective communication skills serve as a key component of excellent veterinary care and provide a foundation for building trusting relationships with clients. While many veterinary clients value their pets for companionship, the focus of other relationships may be based on a partnership between the human and animal, as is the case with the handlers of service dogs. As the use of service dogs in the US continues to grow, it is important that veterinary professionals are educated on how best to meet the unique needs of service dogs and their handlers. This article evaluates the interactions of veterinary students with a service dog handler in a simulated client scenario. Ten videotaped interactions were coded to assess third-year students' communication skills (nonverbal communication, open-ended questions, reflective listening, and empathy), and their ability to discuss the diagnostic and therapeutic options for a dog with suspected intervertebral disk disease. Results showed that the majority of students demonstrated competence in the use of nonverbal communication skills and in discussing the biomedical aspects of the disease. Students require development in the use of open-ended questions, reflective listening statements, and expression of empathy, as well as building client rapport and discussing the psychosocial aspect of the disease on the client and patient. These findings suggest that veterinary students may benefit from targeted instruction on "best practices" in caring for service dogs and their handlers, including greater attention to the psychosocial aspects of a disease, and from additional communication practice using standardized clients with service dogs.


Mutational analysis of the human DNA polymerase alpha. The most conserved region in alpha-like DNA polymerases is involved in metal-specific catalysis.
Five site-directed mutations were introduced at the most conserved amino acids in region I (YGDTDS) of the human DNA polymerase alpha catalytic subunit. Mutant proteins were expressed in the baculovirus system by an improved method and purified by a rapid one-step purification in high yield and high specific activity. The Asp1004 to Asn mutation produced a protein with no detectable polymerase activity while other mutations gave activities from 1 to 20% of the wild type polymerase activity. Steady state kinetic analysis of the active mutants indicates that none of the mutations caused a change in Km(dNTP) or KD(DNA), but all active mutants showed a decrease in kcat. Thus, the effect of these conserved mutations is manifest in altered rates of catalysis. Two mutations, Asp1002 to Asn and Thr1003 to Ser, caused the enzyme to utilize Mn2+ more effectively in catalysis than Mg2+, suggesting that these amino acids are involved in metal binding. Rates of catalysis by the D1002N and T1003S mutants, as well as Y1000F mutant were improved 80-, 30-, and 70-fold, respectively, on homopolymer templates when Mn2+ replaced Mg2+ as the activator metal. The results from these mutational studies suggest that this highly conserved region binds the metal which is essential for catalysis. The Asp1002 may participate directly in chelating the metal. Results from the T1003S mutant suggest that the beta-methyl group of the threonine side chain might be locked in a hydrophobic pocket preventing free rotation around the C alpha-C beta bond, thus positioning the Thr1003 hydroxyl group to form a crucial bond with the metal ion. In addition, D1002N and T1003S displayed a 20-fold resistance to aphidicolin compared to the wild type polymerase alpha, and all of the active mutants displayed altered sensitivity to butylphenyl-dGTP. Models of the involvement of region I in catalysis and aphidicolin interaction are proposed. The mutational studies presented in this report will serve as a prototype for the functional role of region I in catalysis for all alpha-like DNA polymerases.


Control, Norms, and Attitudes: Differences Between Students Who Do and Do Not Intervene as Bystanders to Sexual Assault.
Sexual assault is a major concern on the U.S. college campus. Engaging students as pro-social bystanders has become more common as a potentially effective mechanism for reducing the incidence of sexual assault and mitigating the harm of assaults that have already occurred. Understanding the influences of pro-social bystander behavior is imperative to developing effective programs, and the use of an evidence-based theoretical framework can help identify the differences between students who intervene and those who do not when presented with the opportunity. A sample of 815 undergraduate university students completed the Sexual Assault Bystander Behavior Questionnaire, a survey based on the theory of planned behavior (TPB) that investigates students' perceived behavioral control to intervene, subjective norms that support intervening, attitudes toward intervening, and intent to intervene in the future. Two-tailed t tests revealed interveners reported significantly greater perceived behavioral control than non-interveners for eight of the 12 intervention behaviors, more supportive subjective norms than non-interveners for seven of the 12 intervention behaviors, more positive attitudes than non-interveners for only one of the 12 intervention behaviors, and greater intent to intervene in the future for six of the 12 intervention behaviors. Differences in the four TPB variables were not consistent for the 12 intervention behaviors. The use of a theoretical framework found to be effective in explaining-and changing-other health-related behaviors, and the inquiry into students' opportunities to intervene to compare against their reported intervention behaviors, is new to this body of literature and contributes to the understanding of the influences of pro-social bystander behavior.


Heterogeneity Aware Random Forest for Drug Sensitivity Prediction.
Samples collected in pharmacogenomics databases typically belong to various cancer types. For designing a drug sensitivity predictive model from such a database, a natural question arises whether a model trained on diverse inter-tumor heterogeneous samples will perform similar to a predictive model that takes into consideration the heterogeneity of the samples in model training and prediction. We explore this hypothesis and observe that ensemble model predictions obtained when cancer type is known out-perform predictions when that information is withheld even when the samples sizes for the former is considerably lower than the combined sample size. To incorporate the heterogeneity idea in the commonly used ensemble based predictive model of Random Forests, we propose Heterogeneity Aware Random Forests (HARF) that assigns weights to the trees based on the category of the sample. We treat heterogeneity as a latent class allocation problem and present a covariate free class allocation approach based on the distribution of leaf nodes of the model ensemble. Applications on CCLE and GDSC databases show that HARF outperforms traditional Random Forest when the average drug responses of cancer types are different.


[Closure of wide patent ductus arteriosus using a fenestrated muscular VSD occluder device in a pediatric patient with Down syndrome and pulmonary hypertension].
Patients with wide patent ductus arteriosus and significant pulmonary hypertension not treated in time constitute a significant problem for cardiologists. For these patients, tests that could aid in decision-making for further planning include reversibility and balloon occlusion tests performed in the catheterization laboratory. Devices developed for the closure of ductus as well as different devices with off-label use may be employed in patients scheduled for transcatheter occlusion. When result of reversibility test is borderline positive, the use of fenestrated device may be applicable for selected patients. Presently described is case of a 10-year-old patient with Down syndrome who had a wide ductus and systemic pulmonary hypertension. Transcatheter closure procedure was performed with off-label use of a fenestrated muscular ventricular septal defect occluder device.


Lens Position Parameters as Predictors of Intraocular Pressure Reduction After Cataract Surgery in Nonglaucomatous Patients With Open Angles.
To evaluate the relationship between lens position parameters and intraocular pressure (IOP) reduction after cataract surgery in nonglaucomatous eyes with open angles. The main outcome of the prospective study was percentage of IOP change, which was calculated using the preoperative IOP and the IOP 4 months after cataract surgery in nonglaucomatous eyes with open angles. Lens position (LP), defined as anterior chamber depth (ACD) + 1/2 lens thickness (LT), was assessed preoperatively using parameters from optical biometry. Preoperative IOP, central corneal thickness, ACD, LT, axial length (AXL), and the ratio of preoperative IOP to ACD (PD ratio) were also evaluated as potential predictors of percentage of IOP change. The predictive values of the parameters we found to be associated with the primary outcome were compared. Four months after cataract surgery, the average IOP reduction was 2.03 ± 2.42 mm Hg, a 12.74% reduction from the preoperative mean of 14.5 ± 3.05 mm Hg. Lens position was correlated with IOP reduction percentage after adjusting for confounders (P = 0.002). Higher preoperative IOP, shallower ACD, shorter AXL, and thicker LT were significantly associated with percentage of IOP decrease. Although not statistically significant, LP was a better predictor of percentage of IOP change compared to PD ratio, preoperative IOP, and ACD. The percentage of IOP reduction after cataract surgery in nonglaucomatous eyes with open angles is greater in more anteriorly positioned lenses. Lens position, which is convenient to compute by basic ocular biometric data, is an accessible predictor with considerable predictive value for postoperative IOP change.


The views of stakeholders on the role of the primary school in preventing childhood obesity: a qualitative systematic review.
Schools are increasingly recognized as an ideal setting for interventions to tackle childhood obesity. A better understanding of the views of key stakeholders would help to engage schools and inform the feasibility of such interventions in practice. This meta-synthesis of 18 qualitative studies explores the views of parents, school staff, school governors, school nurses and students on the role of the primary school in preventing childhood obesity. Six categories emerged: 'School as a key setting'; 'What schools should be doing to promote healthy eating (HE)'; 'What schools should be doing to promote physical activity (PA)'; 'General barriers'; 'Barriers to promoting HE at school'; and 'Barriers to promoting PA at school'. Thirty-seven finer-level themes emerged within these categories. Stakeholders agreed on the key role of the primary school as a setting for obesity prevention, the importance of schools providing and promoting opportunities for HE and PA, and the need for schools to work with parents. Some perceived barriers could be overcome at school level, e.g. using unhealthy foods as rewards/fundraisers or withholding PA for bad behaviour. Leadership and guidance from government were considered to be needed to counteract other observed barriers, particularly regarding school canteens, support for parents and time for PA.


Adverse effects in hospice patients with chronic kidney disease receiving hydromorphone.
Pain is one the most common symptoms experienced by palliative care patients. The treatment of pain involves the use of strong opioids such as hydromorphone, morphine, methadone, fentanyl, oxycodone, oxymorphone, or levorphanol for moderate to severe pain. Hydromorphone is metabolized by the liver to hydromorphone-3-glucuronide (H3G), a compound that can potentially cause neuroexcitatory phenomena with accumulation. Pharmacokinetic studies have shown that H3G levels in patients with renal insufficiency are 4 times as high as those with normal renal function; however, reports have been conflicting as to whether or not it is safe to use hydromorphone in renal insufficiency. In this study we sought to determine the prevalence of neuroexcitation in patients with renal insufficiency who were given hydromorphone, as measured by the glomerular filtration rate (GFR), and to investigate factors associated with increased risk of neuroexcitation in this patient group. For the 12- month period from June 2007 through June 2008, charts of inpatient hospice patients that showed a glomerular filtration rate of <60 (mL/min/1.73 m(2)) and hydromorphone administration for pain control via continuous infusion were reviewed for the occurrence of neuroexcitatory effects, including tremor, myoclonus, agitation, cognitive dysfunction, and seizures. Overall prevalence of neuroexcitatory effects were: tremor 11 (20%), myoclonus 11 (20%), agitation 26 (48%), and cognitive dysfunction 21 (39%). No seizures were observed. No neuroexcitatory effects were observed for the lowest quartile of dose or duration of hydromorphone. There was a strong and graded increase in neuroexcitatory effects with increasing quartile of dose or duration of hydromorphone for agitation (dose, p<0.0001; duration, p<0.0001) and cognitive dysfunction (dose, p<0.0002; duration, p<0.002). Consistent but weaker trends were observed for tremor and myoclonus. Parenteral hydromorphone has few neuroexcitatory symptoms until H3G accumulates past a neurotoxic threshold, such as might occur with increasing dose or duration, which, when exceeded, causes neuroexcitatory symptoms to manifest.


Comparative analysis of doublets versus single-layer diffractive optical elements in eyepiece or magnifier design.
We quantify the impact of eye clearance requirement on the performance of eyepieces utilizing doublets versus diffractive optical elements on aspheric substrates. In this study, the doublets were designed to be cemented on-axis elements. Specifically, four different values of eye clearance were implemented: 17, 20, 23, and 26 mm. For each value, axial and lateral color, spherical aberration, coma, astigmatism, field curvature, and distortion were compared. Each system under comparison was optimized for the same focal length, a 9 mm exit pupil, photopic wavelengths (513-608 nm), and a 40 degrees full field of view. We demonstrate that the single-layer diffractive optical element supports an eye clearance value of approximately 80% of the effective focal length, while the doublet drops below desired specifications at approximately 65% of the effective focal length.


Systematic comparison of microarray profiling, real-time PCR, and next-generation sequencing technologies for measuring differential microRNA expression.
RNA abundance and DNA copy number are routinely measured in high-throughput using microarray and next-generation sequencing (NGS) technologies, and the attributes of different platforms have been extensively analyzed. Recently, the application of both microarrays and NGS has expanded to include microRNAs (miRNAs), but the relative performance of these methods has not been rigorously characterized. We analyzed three biological samples across six miRNA microarray platforms and compared their hybridization performance. We examined the utility of these platforms, as well as NGS, for the detection of differentially expressed miRNAs. We then validated the results for 89 miRNAs by real-time RT-PCR and challenged the use of this assay as a "gold standard." Finally, we implemented a novel method to evaluate false-positive and false-negative rates for all methods in the absence of a reference method.


Analysis of the vaccine-induced influenza B virus hemagglutinin-specific antibody dependent cellular cytotoxicity response.
Influenza A virus (IAV) and influenza B virus (IBV) cause substantial morbidity and mortality during seasonal epidemics. On basis of variation in the surface glycoprotein hemagglutinin, two antigenically distinct lineages of IBV are distinguished: B/Victoria/2/87-like (B/Vic) and B/Yamagata/16/88-like (B/Yam). To prevent IAV and IBV infections, both trivalent (containing IBV of one lineage) and quadrivalent (containing IBV of both lineages) influenza vaccines are used. In addition to virus-neutralizing antibodies, inactivated influenza vaccines induce antibodies that mediate antibody-dependent cellular cytotoxicity (ADCC). Here, we determine whether vaccination with trivalent or quadrivalent inactivated influenza vaccine induces ADCC mediating antibodies directed to IBV of the two different lineages, and whether these antibodies cross-react with IBV of the opposing lineage. A robust ADCC assay based on the use of recombinant hemagglutinin and a continuous natural killer cell line that expresses FcγRIII (CD16) was used to detect the presence of ADCC mediating antibodies. Paired pre- and post-vaccination serum samples from 26 and 15 study subjects that received a trivalent or quadrivalent inactivated influenza vaccine, respectively, were assessed for the presence of ADCC mediating antibodies specific for HA derived from viruses of the B/Vic or B/Yam-lineage. Furthermore, the relative contribution of HA1- and HA2-subunit-specific antibodies to the ADCC response was determined. We found that seasonal inactivated influenza vaccines induce HA-head- and HA-stalk-specific antibodies that mediate ADCC. As expected, the quadrivalent vaccine induced antibodies to HA from both IBV lineages. Notably, a trivalent vaccine containing HA from the B/Vic lineage induced antibodies that cross-react with the B/Yam lineage.


Requirement for genes with homology to ABC transport systems for attachment and virulence of Agrobacterium tumefaciens.
Transposon mutants of Agrobacterium tumefaciens which were avirulent and unable to attach to plant cells were isolated and described previously. A clone from a library of Agrobacterium tumefaciens DNA which was able to complement these chromosomal att mutants was identified. Tn3HoHo1 insertions in this clone were made and used to replace the wild-type genes in the bacterial chromosome by marker exchange. The resulting mutants were avirulent and showed either no or very much reduced attachment to carrot suspension culture cells. We sequenced a 10-kb region of this clone and found a putative operon containing nine open reading frames (ORFs) (attA1A2BCDEFGH). The second and third ORFs (attA2 and attB) showed homology to genes encoding the membrane-spanning proteins (potB and potH; potC and potI) of periplasmic binding protein-dependent (ABC) transport systems from gram-negative bacteria. The homology was strongest to proteins involved in the transport of spermidine and putrescine. The first and fifth ORFs (attA1 and attE) showed homology to the genes encoding ATP-binding proteins of these systems including potA, potG, and cysT from Escherichia coli; occP from A. tumefaciens; cysA from Synechococcus spp.; and ORF-C from an operon involved in the attachment of Campylobacte jejuni. The ability of mutants in these att genes to bind to host cells was restored by addition of conditioned medium during incubation of the bacteria with host cells.


Genetic relationships and evolution of genotypes of yellow fever virus and other members of the yellow fever virus group within the Flavivirus genus based on the 3' noncoding region.
Genetic relationships among flaviviruses within the yellow fever (YF) virus genetic group were investigated by comparing nucleotide sequences of the 3' noncoding region (3'NCR). Size heterogeneity was observed between members and even among strains of the same viral species. Size variation between YF strains was due to duplications and/or deletions of repeated nucleotide sequence elements (RYF). West African genotypes had three copies of the RYF (RYF1, RYF2, and RYF3); the Angola and the East and Central African genotypes had two copies (RYF1 and RYF3); and South American genotypes had only a single copy (RYF3). Nucleotide sequence analyses suggest a deletion within the 3'NCR of South American genotypes, including RYF1 and RYF2. Based on studies with the French neurotropic vaccine strain, passage of a YF virus strain in cell culture can result in deletion of RYF1 and RYF2. Taken together, these observations suggest that South American genotypes of YF virus evolved from West African genotypes and that the South American genotypes lost RYF1 and RYF2, possibly in a single event. Repeated sequence elements were found within the 3'NCR of other members of the YF virus genetic group, suggesting that it is probably characteristic for members of the YF virus genetic group. A core sequence of 15 nucleotides, containing two stem-loops, was found within the 3'NCR of all members of the YF genetic group and may represent the progenitor repeat sequence. Secondary structure predictions of the 3'NCR showed very similar structures for viruses that were closely related phylogenetically.


Influence of doping on thermal diffusivity of single crystals used in photonics: measurements based on thermal wave methods.
Three crystals used in solid-state lasers, namely, yttrium aluminum garnet (YAG), yttrium orthovanadate (YVO(4)), and gadolinium calcium oxoborate (GdCOB), were investigated to determine the influence of dopants on their thermal diffusivity. The thermal diffusivity was measured by thermal wave method with a signal detection based on mirage effect. The YAG crystals were doped with Yb or V, the YVO(4) with Nd or Ca and Tm, and the GdCOB crystals contained Nd or Yb. In all cases, the doping caused a decrease in thermal diffusivity. The analysis of complementary measurements of ultrasound velocity changes caused by dopants leads to the conclusion that impurities create phonon scattering centers. This additional scattering reduces the phonon mean free path and accordingly results in the decrease of the thermal diffusivity of the crystal. The influence of doping on lattice parameters was investigated, additionally.


Influence of the substrate lattice structure on the formation of quantum well states in thin In and Pb films on silicon.
The substrate lattice structure may have a considerable influence on the formation of quantum well states in a metal overlayer material. Here we study three model systems using angle resolved photoemission and low energy electron diffraction: indium films on Si(111) and indium and lead on Si(100). Data are compared with theoretical predictions based on density functional theory. We find that the interaction between the substrate and the overlayer strongly influences the formation of quantum well states; indium layers only exhibit well defined quantum well states when the layer relaxes from an initial face-centred cubic to the bulk body-centred tetragonal lattice structure. For Pb layers on Si(100) a change in growth orientation inhibits the formation of quantum well states in films thicker than 2 ML.


Mechanism of phosphorylation-dependent binding of APC to beta-catenin and its role in beta-catenin degradation.
The transcriptional coactivator beta-catenin mediates Wnt growth factor signaling. In the absence of a Wnt signal, casein kinase 1 (CK1) and glycogen synthase kinase-3beta (GSK-3beta) phosphorylate cytosolic beta-catenin, thereby flagging it for recognition and destruction by the ubiquitin/proteosome machinery. Phosphorylation occurs in a multiprotein complex that includes the kinases, beta-catenin, axin, and the Adenomatous Polyposis Coli (APC) protein. The role of APC in this process is poorly understood. CK1epsilon and GSK-3beta phosphorylate APC, which increases its affinity for beta-catenin. Crystal structures of phosphorylated and nonphosphorylated APC bound to beta-catenin reveal a phosphorylation-dependent binding motif generated by mutual priming of CK1 and GSK-3beta substrate sequences. Axin is shown to act as a scaffold for substrate phosphorylation by these kinases. Phosphorylated APC and axin bind to the same surface of, and compete directly for, beta-catenin. The structural and biochemical data suggest a novel model for how APC functions in beta-catenin degradation.


Playing the electric light orchestra--how electrical stimulation of visual cortex elucidates the neural basis of perception.
Vision research has the potential to reveal fundamental mechanisms underlying sensory experience. Causal experimental approaches, such as electrical microstimulation, provide a unique opportunity to test the direct contributions of visual cortical neurons to perception and behaviour. But in spite of their importance, causal methods constitute a minority of the experiments used to investigate the visual cortex to date. We reconsider the function and organization of visual cortex according to results obtained from stimulation techniques, with a special emphasis on electrical stimulation of small groups of cells in awake subjects who can report their visual experience. We compare findings from humans and monkeys, striate and extrastriate cortex, and superficial versus deep cortical layers, and identify a number of revealing gaps in the 'causal map' of visual cortex. Integrating results from different methods and species, we provide a critical overview of the ways in which causal approaches have been used to further our understanding of circuitry, plasticity and information integration in visual cortex. Electrical stimulation not only elucidates the contributions of different visual areas to perception, but also contributes to our understanding of neuronal mechanisms underlying memory, attention and decision-making.


The Association of Peripheral Blood Regulatory T-Cell Concentrations With Epithelial Ovarian Cancer: A Brief Report.
There is a mounting body of evidence demonstrating higher percentages of regulatory T (Treg) cells in the peripheral blood of patients with cancer in comparison to healthy controls, but there is a paucity of epidemiological literature characterizing circulating Treg cells among patients with epithelial ovarian cancer (EOC). To investigate the role of peripheral Treg cells in ovarian neoplasms, we conducted a case-control study to characterize circulating concentrations of Treg cells among patients with EOC, women with benign ovarian conditions, and healthy controls without a history of cancer. Participants were identified for inclusion due to their participation in the Data Bank and BioRepository program at Roswell Park Cancer Institute in Buffalo, NY. Patients included 71 women with a primary diagnosis of EOC and 195 women with a diagnosis of benign ovarian conditions. Controls included 101 age- and race-matched women without a history of cancer. Nonfasting, pretreatment peripheral blood levels of CD3+CD4+CD25+FOXP3+ Treg cells were measured using flow cytometric analyses and expressed as a percentage of total CD3+ cells and as a percentage of total CD3+CD4+ cells. Compared to healthy controls and women with benign ovarian conditions, patients with EOC had significantly higher frequency of Treg cells (P < 0.04). In multivariable logistic regression analyses using Treg frequency expressed as a percentage of CD+3 cells, we observed a significant positive association between Treg cell percentage and EOC risk, with each 1% increase associated with a 37% increased risk of EOC (odds ratio, 1.37; 95% confidence interval, 1.04-1.80). We observed a similar trend when Treg frequency was expressed as a percentage of CD3+CD+4 cells (odds ratio, 1.22; 95% confidence interval, 0.99-1.49). The current study provides support that peripheral Treg cell frequency is elevated in patients with EOC in comparison to women with benign ovarian conditions and healthy controls.