Characterisation of the transcriptome of male and female wild-type guppy brains with RNA-Seq and consequences of exposure to the pharmaceutical pollutant, 17α-ethinyl estradiol.
Waterways are increasingly being contaminated by chemical compounds that can disrupt the endocrinology of organisms. One such compound is 17α-ethinyl estradiol (EE2), a synthetic estrogen used in the contraceptive pill. Despite considerable research interest in the effects of EE2 on reproduction and gene expression, surprisingly, only a few studies have capitalised on technologies, such as next-generation sequencing (NGS), to uncover the molecular pathways related to EE2 exposure. Accordingly, using high-throughput sequencing technologies, the aim of our study was to explore the effects of EE2 on brain transcriptome in wild-type male and female guppy (Poecilia reticulata). We conducted two sets of experiments, where fish were exposed to EE2 (measured concentrations: 8ng/L and 38ng/L) in a flow-through system for 21days. The effects on the brain transcriptome on both males and females were assessed using Illumina sequencing (MiSeq and HiSeq) platform followed by bioinformatics analysis (edgeR, DESeq2). Here, we report that exposure to EE2 caused both up- and downregulation of specific transcript abundances, and affected transcript abundance in a sex-specific manner. Specifically, we found 773 transcripts, of which 60 were male-specific, 61 female-specific and 285 treatment-specific. EE2 affected expression of 165 transcripts in males, with 88 downregulated and 77 upregulated, while in females, 120 transcripts were affected with 62 downregulated and 58 upregulated. Finally, RT-qPCR validation demonstrated that expression of transcripts related to transposable elements, neuroserpin and heat shock protein were significantly affected by EE2-exposure. Our study is the first to report brain transcriptome libraries for guppies exposed to EE2. Not only does our study provide a valuable resource, it offers insights into the mechanisms underlying the feminizing effects on the brains of organisms exposed to environmentally realistic concentrations of EE2.


Internet consultations from a remote Pacific island: impact of digitized radiologic images on referral decisions.
A study was carried out to determine whether digitized radiologic images added valuable information to Internet consultations from a remote Pacific Island. Chuuk State Hospital (Federated States of Micronesia) has limited film screen radiology, minimal ultrasound capability, and no radiologist. Providers initiate Web-based referrals for consultation or patient transfer. Digitized images (via low-cost digital camera or flatbed scanner) were uploaded to a Web site. Images were assessed for impact on referral decisions. A radiologist scored image quality and confidence (scale: 1-7). Of 97 referrals with images that were reviewed, 74 (76%) image sets were abnormal, 20 (20%) were normal, and 3 (4%) were indeterminate. Median scores were 4 for image quality and 5 for diagnostic confidence. In most cases with abnormal radiology (52/74, 70%), images were considered valuable. Radiologic images digitized with a low-cost camera or flatbed scanner provided valuable information for decision making in an Internet-based consultation and referral process from a remote, impoverished Pacific Island jurisdiction, despite relatively low image quality.


Assessing the sustained impact of a school-based obesity prevention program for adolescent boys: the ATLAS cluster randomized controlled trial.
Obesity prevention interventions targeting 'at-risk' adolescents are urgently needed. The aim of this study is to evaluate the sustained impact of the 'Active Teen Leaders Avoiding Screen-time' (ATLAS) obesity prevention program. Cluster RCT in 14 secondary schools in low-income communities of New South Wales, Australia. Participants were 361 adolescent boys (aged 12-14 years) 'at risk' of obesity. The intervention was based on Self-Determination Theory and Social Cognitive Theory and involved: professional development, fitness equipment for schools, teacher-delivered physical activity sessions, lunch-time activity sessions, researcher-led seminars, a smartphone application, and parental strategies. Assessments for the primary (body mass index [BMI], waist circumference) and secondary outcomes were conducted at baseline, 8- (post-intervention) and 18-months (follow-up). Analyses followed the intention-to-treat principle using linear mixed models. After 18-months, there were no intervention effects for BMI or waist circumference. Sustained effects were found for screen-time, resistance training skill competency, and motivational regulations for school sport. There were no clinically meaningful intervention effects for the adiposity outcomes. However, the intervention resulted in sustained effects for secondary outcomes. Interventions that more intensively target the home environment, as well as other socio-ecological determinants of obesity may be needed to prevent unhealthy weight gain in adolescents from low-income communities. Australian Clinical Trial Registry ACTRN12612000978864.


Prognostic Value of Human Peripheral Monocyte Subsets for Future Coronary Events in Patients Without Significant Coronary Artery Stenosis.
Monocytes in human peripheral blood are heterogeneous and can be divided into 2 groups, inflammatory and pro-inflammatory, according to the differential expression of CD14 and CD16. Pro-inflammatory monocytes (CD14+CD16+) seem to contribute to the development of coronary artery disease. This study aimed to investigate the involvement of specific human peripheral monocyte subsets in the development of future coronary events.Methods and Results:We enrolled 271 patients who were suspected to have either stable angina pectoris or silent myocardial ischemia and underwent coronary angiography (CAG). Two monocyte subsets (CD14+CD16-and CD14+CD16+) were measured by flow cytometry. Patients who did not undergo coronary artery revascularization at initial CAG were followed as the medical therapy group, which included 136 patients among whom 15 had future coronary events. The frequency of CD14+CD16+monocytes was significantly higher in patients who had future coronary events than in those who did not (P<0.01). Furthermore, the frequencies of CD14+CD16+monocyte were not significantly different between patients who had future coronary events and those who underwent coronary revascularization at initial CAG (P<0.33). Multivariate analysis revealed that the frequency of CD14+CD16+monocytes was an independent predictor for future coronary events (P<0.01). An increase in the abundance of human peripheral pro-inflammatory monocytes is related to the development of future coronary events.


Interpreting genetic variability: the effects of shared evolutionary history.
Data from different individuals at a single locus are positively correlated because of the shared genealogy of the sampled genes. This paper illustrates the qualitative effects on genealogical trees of assumptions about population demography, and it considers the consequences for genetic variability. An understanding of these effects is invaluable in the interpretation of data and for inferences about population history. In contrast, traditional genetic measures of diversity and approximation methods do not seem well suited for addressing the problem.


Capgras' syndrome in first-episode psychotic disorders.
Misidentification phenomena, including the delusion of 'imposters' named after Joseph Capgras, occur in various major psychiatric and neurological disorders but have rarely been studied systematically in broad samples of modern patients. This study investigated the prevalence and correlated clinical factors of Capgras' phenomenon in a broad sample of patient-subjects with first-lifetime episodes of psychotic affective and nonaffective disorders. We evaluated 517 initially hospitalized, first-episode psychotic-disorder patients for the prevalence of Capgras' phenomenon and its association with DSM-IV-TR diagnoses including schizophreniform, brief psychotic, unspecified psychotic, delusional, and schizoaffective disorders, schizophrenia, bipolar-I disorder and major depression with psychotic features, and with characteristics of interest including antecedent psychiatric and neurological morbidity, onset type and presenting psychopathological phenomena, using standard bivariate and multivariate statistical methods. Capgras' syndrome was identified in 73/517 (14.1%) patients (8.2-50% across diagnoses). Risk was greatest with acute or brief psychotic disorders (schizophreniform psychoses 50%, brief psychoses 34.8%, or unspecified psychoses 23.9%), intermediate in major depression (15%), schizophrenia (11.4%) and delusional disorder (11.1%), and lowest in bipolar-I (10.3%) and schizoaffective disorders (8.2%). Associated were somatosensory, olfactory and tactile hallucinations, Schneiderian (especially delusional perception), and cycloid features including polymorphous psychotic phenomena, rapidly shifting psychomotor and affective symptoms, pananxiety, ecstasy, overconcern with death, and perplexity or confusion, as well as rapid onset, but not sex, age, abuse history, dissociative features, or indications of neurological disorders. Capgras' syndrome was prevalent across a broad spectrum of first-episode psychotic disorders, most often in acute psychoses of rapid onset.


Only five of 10 strictly conserved disulfide bonds are essential for folding and eight for function of the HIV-1 envelope glycoprotein.
Protein folding in the endoplasmic reticulum goes hand in hand with disulfide bond formation, and disulfide bonds are considered key structural elements for a protein's folding and function. We used the HIV-1 Envelope glycoprotein to examine in detail the importance of its 10 completely conserved disulfide bonds. We systematically mutated the cysteines in its ectodomain, assayed the mutants for oxidative folding, transport, and incorporation into the virus, and tested fitness of mutant viruses. We found that the protein was remarkably tolerant toward manipulation of its disulfide-bonded structure. Five of 10 disulfide bonds were dispensable for folding. Two of these were even expendable for viral replication in cell culture, indicating that the relevance of these disulfide bonds becomes manifest only during natural infection. Our findings refine old paradigms on the importance of disulfide bonds for proteins.


How do glucocorticoids influence stress responses? Integrating permissive, suppressive, stimulatory, and preparative actions.
The secretion of glucocorticoids (GCs) is a classic endocrine response to stress. Despite that, it remains controversial as to what purpose GCs serve at such times. One view, stretching back to the time of Hans Selye, posits that GCs help mediate the ongoing or pending stress response, either via basal levels of GCs permitting other facets of the stress response to emerge efficaciously, and/or by stress levels of GCs actively stimulating the stress response. In contrast, a revisionist viewpoint posits that GCs suppress the stress response, preventing it from being pathologically overactivated. In this review, we consider recent findings regarding GC action and, based on them, generate criteria for determining whether a particular GC action permits, stimulates, or suppresses an ongoing stress-response or, as an additional category, is preparative for a subsequent stressor. We apply these GC actions to the realms of cardiovascular function, fluid volume and hemorrhage, immunity and inflammation, metabolism, neurobiology, and reproductive physiology. We find that GC actions fall into markedly different categories, depending on the physiological endpoint in question, with evidence for mediating effects in some cases, and suppressive or preparative in others. We then attempt to assimilate these heterogeneous GC actions into a physiological whole.


Psychometric evaluation of the Orofacial Pain Scale for Non-Verbal Individuals as a screening tool for orofacial pain in people with dementia.
The aim of this study was to describe the psychometric evaluation of the Orofacial Pain Scale for Non-Verbal Individuals (OPS-NVI) as a screening tool for orofacial pain in people with dementia. The OPS-NVI has recently been developed and needs psychometric evaluation for clinical use in people with dementia. The pain self-report is imperative as a reference standard and can be provided by people with mild-to-moderate cognitive impairment. The presence of orofacial pain during rest, drinking, chewing and oral hygiene care was observed in people with mild cognitive impairment (MCI) and dementia using the OPS-NVI. Participants who were considered to present a reliable self-report were asked about pain presence, and in all participants, the oral health was examined by a dentist for the presence of potential painful conditions. After item-reduction, inter-rater reliability and criterion validity were determined. The presence of orofacial pain in this population was low (0%-10%), resulting in an average Positive Agreement of 0%-100%, an average Negative Agreement of 77%-100%, a sensitivity of 0%-100% and a specificity of 66%-100% for the individual items of the OPS-NVI. At the same time, the presence of oral problems, such as ulcers, tooth root remnants and caries was high (64.5%). The orofacial pain presence in this MCI and dementia population was low, resulting in low scores for average Positive Agreement and sensitivity and high scores for average Negative Agreement and specificity. Therefore, the OPS-NVI in its current form cannot be recommended as a screening tool for orofacial pain in people with MCI and dementia. However, the inter-rater reliability and criterion validity of the individual items in this study provide more insight for the further adjustment of the OPS-NVI for diagnostic use. Notably, oral health problems were frequently present, although no pain was reported or observed, indicating that oral health problems cannot be used as a new reference standard for orofacial pain, and a regular oral examination by care providers and oral hygiene care professionals remains indispensable.


Ocular gene transfer in the spotlight: implications of newspaper content for clinical communications.
Ocular gene transfer clinical trials are raising hopes for blindness treatments and attracting media attention. News media provide an accessible health information source for patients and the public, but are often criticized for overemphasizing benefits and underplaying risks of novel biomedical interventions. Overly optimistic portrayals of unproven interventions may influence public and patient expectations; the latter may cause patients to downplay risks and over-emphasize benefits, with implications for informed consent for clinical trials. We analyze the news media communications landscape about ocular gene transfer and make recommendations for improving communications between clinicians and potential trial participants in light of media coverage. We analyzed leading newspaper articles about ocular gene transfer (1990-2012) from United States (n = 55), Canada (n = 26), and United Kingdom (n = 77) from Factiva and Canadian Newsstand databases using pre-defined coding categories. We evaluated the content of newspaper articles about ocular gene transfer for hereditary retinopathies, exploring representations of framing techniques, research design, risks/benefits, and translational timelines. The dominant frame in 61% of stories was a celebration of progress, followed by human-interest in 30% of stories. Missing from the positive frames were explanations of research design; articles conflated clinical research with treatment. Conflicts-of-interest and funding sources were similarly omitted. Attention was directed to the benefits of gene transfer, while risks were only reported in 43% of articles. A range of visual outcomes was described from slowing vision loss to cure, but the latter was the most frequently represented even though it is clinically infeasible. Despite the prominence of visual benefit portrayals, 87% of the articles failed to provide timelines for the commencement of clinical trials or for clinical implementation. Our analysis confirms that despite many initiatives to improve media communications about experimental biotechnologies, media coverage remains overly optimistic and omits important information. In light of these findings, our recommendations focus on the need for clinicians account for media coverage in their communications with patients, especially in the context of clinical trial enrolment. The development of evidence-based communication strategies will facilitate informed consent and promote the ethical translation of this biotechnology.


Epidemiology of Acne Vulgaris in 18-Year-Old Male Army Conscripts in a South Brazilian City.
Prevalence of acne varies worldwide. Several factors (age, skin color, body fat, diet, and smoking) have been investigated as risk factors. A total of 2,201 18-year-old males living in Pelotas, South Brazil, were evaluated in order to examine the prevalence of acne and associated factors. A cross-sectional population-based study was conducted. A dermatologist performed the clinical examination of the face and trunk for identification of acne lesions. Acne was evaluated as clinically noninflammatory, inflammatory, and acne with both types of lesions. Skin color, schooling, height, smoking, skinfolds, waist circumference, BMI, and dietary dairy intake were the independent variables used. A response rate of 97.2% was obtained. Individuals without any acne lesion were 241 (10.9%); 161 (7.3%) only had noninflammatory lesions, 404 (18.4%) only inflammatory lesions; and 1,395 (63.4%) presented both types of lesions. In multivariate analysis, the type of lesions was different in light and dark skin phototype adolescents, with more common inflammatory lesions in the light phototype and noninflammatory ones in the dark phototype patients. Height was directly associated with the occurrence of all types of acne, whereas lower fat mass was associated with the occurrence of noninflammatory acne. While daily consumption of whole milk or yogurt was found to be associated with inflammatory acne in crude analysis, the association with milk was not detected and that with yogurt was low in multivariate analysis. Our results suggest that future studies should explore determinants of noninflammatory and inflammatory acne separately, especially if mixed populations are studied.


FOXA2 regulates a network of genes involved in critical functions of human intestinal epithelial cells.
The forkhead box A (FOXA) family of pioneer transcription factors is critical for the development of many endoderm-derived tissues. Their importance in regulating biological processes in the lung and liver is extensively characterized, though much less is known about their role in intestine. Here we investigate the contribution of FOXA2 to coordinating intestinal epithelial cell function using postconfluent Caco2 cells, differentiated into an enterocyte-like model. FOXA2 binding sites genome-wide were determined by ChIP-seq and direct targets of the factor were validated by ChIP-qPCR and siRNA-mediated depletion of FOXA1/2 followed by RT-qPCR. Peaks of FOXA2 occupancy were frequent at loci contributing to gene ontology pathways of regulation of cell migration, cell motion, and plasma membrane function. Depletion of both FOXA1 and FOXA2 led to a significant reduction in the expression of multiple transmembrane proteins including ion channels and transporters, which form a network that is essential for maintaining normal ion and solute transport. One of the targets was the adenosine A2B receptor, and reduced receptor mRNA levels were associated with a functional decrease in intracellular cyclic AMP. We also observed that 30% of FOXA2 binding sites contained a GATA motif and that FOXA1/A2 depletion reduced GATA-4, but not GATA-6 protein levels. These data show that FOXA2 plays a pivotal role in regulating intestinal epithelial cell function. Moreover, that the FOXA and GATA families of transcription factors may work cooperatively to regulate gene expression genome-wide in the intestinal epithelium.


Microsomal enzymes in malnutrition as determined by plasma half life of antipyrine.
Nutritional-pharmacological interactions were studied in a group of malnourished subjects. Antipyrine was used to evaluate mixed-function oxidase in man. The results indicated that the rate of disappearance of antipyrine from plasma was strongly influenced by the nutritional status of the individual. The half life of antipyrine was modified in undernourished subjects and those with nutritional oedema. This finding indicates that drug regimens may have to be adjusted in patients who have antipyrine half lives that are shorter or longer than normal. Otherwise drug treatment may be inadequate or, in patients with impaired microsomal enzyme activity, potentially dangerous.


The Very Low-Dose Dexamethasone Suppression Test in the General Population: A Cross-Sectional Study.
Determinants of the hypothalamic-pituitary-adrenal (HPA) axis functioning are increasingly explored in population-based studies. However, functional tests measuring the negative feedback of the HPA axis cannot easily be implemented into large observational studies. Furthermore, high doses of dexamethasone often completely suppress the HPA axis in healthy persons. This study aimed to detect the effects of the health, lifestyle and sociodemographic factors, psychiatric problems and cognitive functions on the negative feedback of the HPA axis using a very low-dose (0.25 mg) dexamethasone suppression test (DST).We evaluated the associations of several determinants with the saliva cortisol concentrations after dexamethasone intake in a confounder-adjusted model also corrected for baseline saliva cortisol concentrations in the Rotterdam Study, a large population-based study (N = 1822). We found that female sex, low income, lack of exercise, instrumental disability and smoking were all independently associated with stronger suppression of the HPA axis. Even though there were no linear associations between psychiatric measures and cortisol suppression, we found that depressive symptoms and anxiety disorders were more common in persons with non-suppression of cortisol. Conversely, psychotropic medication use was related to enhanced suppression of cortisol after DST. In this large study, we found that female gender, low socioeconomic status and poor health were all related to suppression of the HPA axis. Non-linear associations were detected between the suppression of the HPA axis and common psychiatric disorders in community-dwelling persons.


Effects of intentional weight loss in patients with obesity and heart failure: a systematic review.
Obesity is an independent risk factor for the development of heart failure, and the two commonly co-exist. The European Society of Cardiology does not provide guidance regarding weight loss strategies in heart failure. The aim of this study was to systematically review the evidence for outcomes following intentional weight loss in patients with heart failure and obesity. A systematic review of English articles was undertaken using databases PubMed, Embase and CENTRAL. Randomized controlled trials and observational studies reporting outcomes following intentional weight loss by lifestyle, surgical or pharmacotherapy intervention in patients with obesity and heart failure were included. Four randomized controlled trials and seven observational studies were identified. Two randomized controlled trials used diet and exercise as an intervention, one used diet alone and one used a pharmacological intervention (orlistat). All but one reported significant weight loss. Two reported improvement in exercise capacity and quality of life. One reported improvement in New York Heart Association functional class in heart failure with preserved ejection fraction. The observational studies, five of which reported on outcomes following bariatric surgery, despite being small, heterogeneous and high risk of bias, suggested a trend in improvement of left ventricular function, quality of life and exercise capacity following weight loss. Weight loss is achievable with lifestyle intervention in those with heart failure and obesity and may result in improvements in New York Heart Association classification, quality of life and exercise capacity.


The AMP-activated protein kinase: role in regulation of skeletal muscle metabolism and insulin sensitivity.
Over the past decade, an epidemic of obesity has developed throughout the Western World. In recent years, significant interest has focused on the role of the AMP-activated protein kinase (AMPK) as a potential therapeutic target for the treatment of obesity and type 2 diabetes and is such the focus of this review. Specifically, the potential role of AMPK in skeletal muscle metabolism as it relates to the insulin sensitizing effects of exercise and the hormones, leptin, adiponectin, ciliary neurotrophic factor and interleukin-6 are discussed. We caution that despite the convincing associations between the activation of AMPK signalling and the restoration of insulin sensitivity, future studies in genetic models of AMPK deficiency or constitutive activation within skeletal muscle are needed to evaluate the quantitative role of AMPK and to validate whether strategies designed to activate skeletal muscle AMPK may be important for regulating whole-body insulin sensitivity.


Synthesis of (±)-amathaspiramide F and discovery of an unusual stereocontrolling element for the [2,3]-Stevens rearrangement.
A formal total synthesis of (±)-amathaspiramide F through a tandem palladium-catalyzed allylic amination/[2,3]-Stevens rearrangement is reported. The unexpected diastereoselectivity of the [2,3]-Stevens rearrangement was controlled by the substitution patterns of an aromatic ring. This discovery represents a new stereocontrolling element for [2,3]-sigmatropic rearrangements in complex molecular settings.


Anxiety, depression, and oscillatory dynamics in a social interaction model.
Although anxiety and depression frequently co-occur and share a substantial part of genetic vulnerability and other risk factors, they are distinct disorders and their effect on social functioning and accompanying cognitive and emotional processing could be different. In this study, in a nonclinical sample, we compared effects of trait anxiety and depressive symptoms on oscillatory dynamics accompanying perception of emotional facial expressions in the context of social interactions. Anxiety was associated with a longer reaction time, with preference of avoidance behavior, and with enhanced event-related alpha desynchronization and diminished theta synchronization. Depression did not show significant behavioral effects and was associated with diminished alpha desynchronization and augmented delta and theta synchronization in prefrontal cortical regions. Thus, in spite of frequent comorbidity, anxiety and depression show opposite patterns of associations with oscillatory dynamics accompanying social interactions. These patterns imply that anxiety is associated with hyper-reactive attentional system, whereas depression show signs of diminished cognitive reactivity. Depression-related enhancement of low-frequency synchronization in prefrontal cortex may reflect a compensatory mechanism of cognitive and emotional upregulation, which depression-prone individuals engage in the process of social interactions.


Green tea extract solutions can control bacterial biofilms formed on space maintainers.
Microorganisms in the mouth are protected from negative environmental conditions by forming biofilms; however, the use of anti-plaque agents in children is not preferred due to toxic side effects. Green tea has been reported to have anti-microbial and anti-dental caries properties. The aim of this study was to assess the ability of green tea extract to prevent the formation of biofilm on the teeth of children using space maintainers. Bacteria were isolated from samples obtained from children aged between 8 and 10 years. The micro-titer plate method and Congo red agar were used to assay biofilm formation. Green tea leaves were obtained from Rize, Turkey. Methanol, hexane and distilled water were used for preparing the extracts. The effects of green tea extract and chlorhexidine on biofilm formation were examined using scanning electron microscopy. Presence of S. mutans 3,3, S. anginosus 2.1.b, S. dysgalactie 22.214.171.124, and E. faecium 10.2. was measured in the biofilm samples. The extracts showed a bacteriostatic effect on the test bacteria, and among the green tea extracts, the methanol extract was found to exhibit the highest efficacy against biofilm formation by S. mutans 3.3. Green tea extract showed good efficacy in controlling bacterial growth, and is recommended as a better-tasting alternative for daily oral hygiene due to a lack of known side effects.


Real and hypothetical rewards.
Laboratory studies of choice and decision making among real monetary rewards typically use smaller real rewards than those common in real life. When laboratory rewards are large, they are almost always hypothetical. In applying laboratory results meaningfully to real-life situations, it is important to know the extent to which choices among hypothetical rewards correspond to choices among real rewards and whether variation of the magnitude of hypothetical rewards affects behavior in meaningful ways. The present study compared real and hypothetical monetary rewards in two experiments. In Experiment 1, participants played a temporal discounting game that incorporates the logic of a repeated prisoner's-dilemma (PD) type game versus tit-for-tat; choice of one alternative ("defection" in PD terminology) resulted in a small-immediate reward; choice of the other alternative ("cooperation" in PD terminology) resulted in a larger reward delayed until the following trial. The larger-delayed reward was greater for half of the groups than for the other half. Rewards also differed in type across groups: multiples of real nickels, hypothetical nickels or hypothetical hundred-dollar bills. All groups significantly increased choice of the larger delayed reward over the 40 trials of the experiment. Over the last 10 trials, cooperation was significantly higher when the difference between larger and smaller hypothetical rewards was greater. Reward type (real or hypothetical) made no significant difference in cooperation. In Experiment 2, real and hypothetical rewards were compared in social discounting - the decrease in value to the giver of a reward as social distance increases to the receiver of the reward. Social discount rates were well described by a hyperbolic function. Discounting rates for real and hypothetical rewards did not significantly differ. These results add to the evidence that results of experiments with hypothetical rewards validly apply in everyday life.


A case of pulmonary oedema induced by suffocation.
A 57-year old female civilian was suffocated by an intruder The victim (experienced severe dyspnoea) but violently resisted the assault. Two hours after this event, on admission to a Trauma Centre, she demonstrated left facial swelling with low percutaneous oxygen saturation. Chest X-ray and computed tomography demonstrated pulmonary oedema. This improved dramatically within a short time and she was discharged on the 5th hospital day. Pulmonary oedema induced by suffocation has been reported only rarely. The possible mechanisms by which pulmonary oedema might form after the relief of airway obstruction are discussed.


Anti-angiogenic pathway associations of the 3p21.3 mapped BLU gene in nasopharyngeal carcinoma.
Zinc-finger, MYND-type containing 10 (ZMYND10), or more commonly called BLU, expression is frequently downregulated in nasopharyngeal carcinoma (NPC) and many other tumors due to promoter hypermethylation. Functional evidence shows that the BLU gene inhibits tumor growth in animal assays, but the detailed molecular mechanism responsible for this is still not well understood. In current studies, we find that 93.5% of early-stage primary NPC tumors show downregulated BLU expression. Using a PCR array, overexpression of the BLU gene was correlated to the angiogenesis network in NPC cells. Moreover, expression changes of the MMP family, VEGF and TSP1, were often detected in different stages of NPC, suggesting the possibility that BLU may be directly involved in the microenvironment and anti-angiogenic activity in NPC development. Compared with vector-alone control cells, BLU stable transfectants, derived from poorly-differentiated NPC HONE1 cells, suppress VEGF165, VEGF189 and TSP1 expression at both the RNA and protein levels, and significantly reduce the secreted VEGF protein in these cells, reflecting an unknown regulatory mechanism mediated by the BLU gene in NPC. Cells expressing BLU inhibited cellular invasion, migration and tube formation. These in vitro results were further confirmed by in vivo tumor suppression and a matrigel plug angiogenesis assay in nude mice. Tube-forming ability was clearly inhibited, when the BLU gene is expressed in these cells. Up to 70-90% of injected tumor cells expressing increased exogenous BLU underwent cell death in animal assays. Overexpressed BLU only inhibited VEGF165 expression in differentiated squamous NPC HK1 cells, but also showed an anti-angiogenic effect in the animal assay, revealing a complicated mechanism regulating angiogenesis and the microenvironment in different NPC cell lines. Results of these studies indicate that alteration of BLU gene expression influences anti-angiogenesis pathways and is important for the development of NPC.


Available extracorporeal treatments for poisoning: overview and limitations.
Poisoning is a significant public health problem. In severe cases, extracorporeal treatments (ECTRs) may be required to prevent or reverse major toxicity. Available ECTRs include intermittent hemodialysis, sustained low-efficiency dialysis, intermittent hemofiltration and hemodiafiltration, continuous renal replacement therapy, hemoperfusion, therapeutic plasma exchange, exchange transfusion, peritoneal dialysis, albumin dialysis, cerebrospinal fluid exchange, and extracorporeal life support. The aim of this article was to provide an overview of the technical aspects, as well as the potential indications and limitations of the different ECTRs used for poisoned patients.


Congenital diaphragmatic hernia candidate genes derived from embryonic transcriptomes.
Congenital diaphragmatic hernia (CDH) is a common (1 in 3,000 live births) major congenital malformation that results in significant morbidity and mortality. The discovery of CDH loci using standard genetic approaches has been hindered by its genetic heterogeneity. We hypothesized that gene expression profiling of developing embryonic diaphragms would help identify genes likely to be associated with diaphragm defects. We generated a time series of whole-transcriptome expression profiles from laser captured embryonic mouse diaphragms at embryonic day (E)11.5 and E12.5 when experimental perturbations lead to CDH phenotypes, and E16.5 when the diaphragm is fully formed. Gene sets defining biologically relevant pathways and temporal expression trends were identified by using a series of bioinformatic algorithms. These developmental sets were then compared with a manually curated list of genes previously shown to cause diaphragm defects in humans and in mouse models. Our integrative filtering strategy identified 27 candidates for CDH. We examined the diaphragms of knockout mice for one of the candidate genes, pre-B-cell leukemia transcription factor 1 (Pbx1), and identified a range of previously undetected diaphragmatic defects. Our study demonstrates the utility of genetic characterization of normal development as an integral part of a disease gene identification and prioritization strategy for CDH, an approach that can be extended to other diseases and developmental anomalies.


Joint effect of multiple common SNPs predicts melanoma susceptibility.
Single genetic variants discovered so far have been only weakly associated with melanoma. This study aims to use multiple single nucleotide polymorphisms (SNPs) jointly to obtain a larger genetic effect and to improve the predictive value of a conventional phenotypic model. We analyzed 11 SNPs that were associated with melanoma risk in previous studies and were genotyped in MD Anderson Cancer Center (MDACC) and Harvard Medical School investigations. Participants with ≥15 risk alleles were 5-fold more likely to have melanoma compared to those carrying ≤6. Compared to a model using the most significant single variant rs12913832, the increase in predictive value for the model using a polygenic risk score (PRS) comprised of 11 SNPs was 0.07(95% CI, 0.05-0.07). The overall predictive value of the PRS together with conventional phenotypic factors in the MDACC population was 0.69 (95% CI, 0.64-0.69). PRS significantly improved the risk prediction and reclassification in melanoma as compared with the conventional model. Our study suggests that a polygenic profile can improve the predictive value of an individual gene polymorphism and may be able to significantly improve the predictive value beyond conventional phenotypic melanoma risk factors.


Responses of a Newly Evolved Auxotroph of Chlamydomonas to B12 Deprivation.
The corrinoid B12 is synthesized only by prokaryotes yet is widely required by eukaryotes as an enzyme cofactor. Microalgae have evolved B12 dependence on multiple occasions, and we previously demonstrated that experimental evolution of the non-B12-requiring alga Chlamydomonas reinhardtii in media supplemented with B12 generated a B12-dependent mutant (hereafter metE7). This clone provides a unique opportunity to study the physiology of a nascent B12 auxotroph. Our analyses demonstrate that B12 deprivation of metE7 disrupts C1 metabolism, causes an accumulation of starch and triacylglycerides, and leads to a decrease in photosynthetic pigments, proteins, and free amino acids. B12 deprivation also caused a substantial increase in reactive oxygen species, which preceded rapid cell death. Survival could be improved without compromising growth by simultaneously depriving the cells of nitrogen, suggesting a type of cross protection. Significantly, we found further improvements in survival under B12 limitation and an increase in B12 use efficiency after metE7 underwent a further period of experimental evolution, this time in coculture with a B12-producing bacterium. Therefore, although an early B12-dependent alga would likely be poorly adapted to coping with B12 deprivation, association with B12-producers can ensure long-term survival whilst also providing a suitable environment for evolving mechanisms to tolerate B12 limitation better.


Reducing mucus production after urinary reconstruction: a prospective randomized trial.
After transposition into the urinary tract, intestinal segments continue to produce mucus. We determine the effectiveness of muco-regulatory drugs, including N-acetylcysteine, aspirin and ranitidine, in reducing mucus secretion and urine viscosity in patients with transposed segments. Our trial was a prospective randomized, double-blind placebo controlled crossover study involving 12 patients who underwent ileal conduit and 31 who underwent bladder reconstruction. Each treatment lasted 3 weeks with a 2-week washout. Pretreatment and posttreatment 24-hour urine samples were analyzed for mucin and viscosity after papain digestion, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and periodic acid-Schiff assay. A disease specific questionnaire and SF-36 quality of life survey were completed. According to the questionnaire, mucus production did not decrease with time in 67% of patients. Mucin comprised 3% of the total nondialyzable material in urine (65 mg./24-hour for ileal conduit and 60 mg./24-hour for bladder reconstruction). Analysis of questionnaires and laboratory results failed to demonstrate any benefit of taking muco-regulatory agents compared with placebo. The use of N-acetylcysteine, aspirin and ranitidine did not result in a reduction in mucin production, urine viscosity or improvement in quality of life.


Molecular analysis of hprt mutations induced by chromium picolinate in CHO AA8 cells.
Chromium picolinate (CrPic) is a popular dietary supplement, marketed to the public for weight loss, bodybuilding, and control of blood sugar. Recommendations for long-term use at high dosages have led to questions regarding its safety. Previous studies have reported that CrPic can cause chromosomal aberrations and mutations. The purpose of the current work was to compare the mutagenicity of CrPic as a suspension in acetone versus a solution in DMSO, and to characterize the hprt mutations induced by CrPic in CHO AA8 cells. Treatments of 2% acetone or 2% DMSO alone produced no significant increase in 6-thioguanine (6-TG)-resistant mutants after 48 h exposures. Mutants resistant to 6-TG were generated by exposing cells for 48 h to 80 microg/cm(2) CrPic in acetone or to 1.0mM CrPic in DMSO. CrPic in acetone produced an average induced mutation frequency (MF) of 56 per 10(6) surviving cells relative to acetone solvent. CrPic in acetone was 3.5-fold more mutagenic than CrPic in DMSO, which produced an MF of 16.2. Characterization of 61 total mutations in 48 mutants generated from exposure to CrPic in acetone showed that base substitutions comprised 33% of the mutations, with transversions being predominant; deletions made up 62% of the mutations, with one-exon deletions predominating; and 1-4 bp insertions made up 5% of the characterized mutations. CrPic induced a statistically greater number of deletions and a statistically smaller number of base substitutions than have been measured in spontaneously generated mutants. These data confirm previous studies showing that CrPic is mutagenic, and support the contention that further study is needed to verify the safety of CrPic for human consumption.


Immunoglobulin G (IgG) responses to Plasmodium falciparum glycosylphosphatidylinositols are short-lived and predominantly of the IgG3 subclass.
The induction of neutralizing immunity to Plasmodium falciparum toxins by vaccination has been proposed as a preventive strategy to limit the severity of malaria. For this approach to be successful, generation of a sustained immune response would be necessary. This study shows that immunoglobulin G (IgG)-subclass responses elicited by the proposed P. falciparum toxin glycosylphosphatidylinositol (GPI) in Papua New Guinean subjects 5-60 years old predominantly involve IgG(3), with a lesser contribution from IgG(1) and an absence of IgG(2) and IgG(4). IgG(3) levels declined sharply within 6 weeks of pharmacological clearance of parasitemia in all subjects, whereas a significant decrease in IgG(1) levels was seen only in subjects < or =19 years old. Because the natural antibody response to P. falciparum GPIs is skewed toward the short-lived IgG(3) subclass, a vaccination strategy with GPI analogues would likely require augmentation by costimulatory molecules, to induce a more persistent anti-GPI response.


Elongation factor-2 kinase regulates autophagy in human glioblastoma cells.
Elongation factor-2 kinase (eEF-2 kinase), also known as Ca(2+)/calmodulin-dependent kinase III, regulates protein synthesis by controlling the rate of peptide chain elongation. The activity of eEF-2 kinase is increased in glioblastoma and other malignancies, yet its role in neoplasia is uncertain. Recent evidence suggests that autophagy plays an important role in oncogenesis and that this can be regulated by mammalian target of rapamycin (mTOR). Because eEF-2 kinase lies downstream of mTOR, we studied the role of eEF-2 kinase in autophagy using human glioblastoma cell lines. Knockdown of eEF-2 kinase by RNA interference inhibited autophagy in glioblastoma cell lines, as measured by light chain 3 (LC3)-II formation, acidic vesicular organelle staining, and electron microscopy. In contrast, overexpression of eEF-2 kinase increased autophagy. Furthermore, inhibition of autophagy markedly decreased the viability of glioblastoma cells grown under conditions of nutrient depletion. Nutrient deprivation increased eEF-2 kinase activity and decreased the activity of S6 kinase, suggesting an involvement of mTOR pathway in the eEF-2 kinase regulation of autophagy. These results suggest that eEF-2 kinase plays a regulatory role in the autophagic process in tumor cells; and eEF-2 kinase is a downstream member of the mTOR signaling; eEF-2 kinase may promote cancer cell survival under conditions of nutrient deprivation through regulating autophagy. Therefore, eEF-2 kinase may be a part of a survival mechanism in glioblastoma and targeting this kinase may represent a novel approach to cancer treatment.


Closed chest cardiac massage in the newborn infant.
The recommended techniques for external cardiac compression during cardiopulmonary resuscitation of the newborn are reviewed. The American Heart Association advises the rescuer to depress the infant's midsternum with the index and forefingers. An alternative recommendation is to encircle the chest with both hands and appose the thumbs on the midsternum. The cases reported here prove the superiority of the latter maneuver over the former in improving cardiac output and systemic blood pressures. The history of external cardiac compression in infants and children is briefly reviewed, further supporting the conclusion that the technique of encircling the chest with thumbs at midsternum is more efficacious and hence the preferred approach to closed chest cardiac massage in the newborn.


Boundary violation ethics: some conceptual clarifications.
The practices of both forensic psychiatry and clinical psychiatry appear to require and to use, in boundary-violation discourse, a special way of referring to the heightened attention to the ethics of interpersonal exchange. But this discourse and the judgments it expresses are each in need of closer scrutiny. A variety of factors make the determination of certain actions to be boundary violations unclear, including the range of alleged boundary-violating behavior, ambiguities in the fundamental metaphor of boundaries violated or transgressed, and confusion about the explanatory status of the value judgments boundary-violation language is used to express. In addition, disputes and disagreements regarding boundary-violation judgments require analysis--an analysis undertaken in this article through appeal to theories of professional role morality. Noted also is the significance of gender in boundary-violation ethics.


Optical effects of thermal structure in the lower atmosphere.
Ascending convective plumes of inhomogeneous warm air interspersed with regions of air that are remarkably free from temperature fluctuations are sometimes observed in the lower layers of the atmosphere. A close correlation is demonstrated between intervals of good optical seeing along an upwardslanting path 20 m long and such periods of below average, air-temperature fluctuation. This correlation is sensitive to the azimuthal angle between wind direction and the vertical plane containing the optical path. The occurrence of temperature-quiescent periods at a given height is also shown to require the horizontal wind speed to be less than a critical value, and there is evidence that the thermal structure tends to be elongated in the direction of the wind. The range of amplitudes of image dancing observed in these experiments is comparable with that encountered in solar observations. The atmospheric temperature structure coefficients calculated from these optical experiments compare favorably with independent direct measurements.


Bone and the 'comforts of life'.
Coffee drinking, smoking and especially alcohol abuse are considered to be risk factors for fractures and osteoporosis. Caffeine causes acute increase in urinary calcium excretion, but epidemiological evidence for the effects of coffee consumption on the risk of fractures is contradictory. Many, (but not all) studies point to decreased bone mass or increased fracture risk in smokers. Alcohol abuse is associated with deleterious changes in bone structure detected by histomorphometry, and with a decrease in bone mineral density (BMD). These changes may also be produced by factors commonly associated with alcohol abuse, e.g. nutritional deficiencies, liver damage and hypogonadism. Alcohol, however, has clear-cut direct effects on bone and mineral metabolism. Acute alcohol intoxication causes transitory hypoparathyroidism with resultant hypocalcaemia and hypercalciuria. As assessed by serum osteocalcin levels, prolonged moderate drinking decreases the function of osteoblasts, the bone-forming cells. In addition, chronic alcoholics are characterized by low serum levels of vitamin D metabolites. Thus, alcohol seems to have a direct toxic effect on bone and mineral metabolism. In contrast, it has recently been reported that moderate alcohol consumption by postmenopausal women may have a beneficial effect on bone.


Whole genome sequencing of two human rhinovirus A types (A101 and A15) detected in Kenya, 2016-2018.
Background: Virus genome sequencing is increasingly utilized in epidemiological surveillance. Genomic data allows comprehensive evaluation of underlying viral diversity and epidemiology to inform control. For human rhinovirus (HRV), genomic amplification and sequencing is challenging due to numerous types, high genetic diversity and inadequate reference sequences. Methods: We developed a tiled amplicon type-specific protocol for genome amplification and sequencing on the Illumina MiSeq platform of two HRV types, A15 and A101. We then assessed added value in analyzing whole genomes relative to the VP4/2 region only in the investigation of HRV molecular epidemiology within the community in Kilifi, coastal Kenya. Results: We processed 73 nasopharyngeal swabs collected between 2016-2018, and 48 yielded at least 70% HRV genome coverage. These included all A101 samples (n=10) and 38 (60.3%) A15 samples. Phylogenetic analysis revealed that the Kilifi A101 sequences interspersed with global A101 genomes available in GenBank collected between 1999-2016. On the other hand, our A15 sequences formed a monophyletic group separate from the global genomes collected in 2008 and 2019. An improved phylogenetic resolution was observed with the genome phylogenies compared to the VP4/2 phylogenies. Conclusions: We present a type-specific full genome sequencing approach for obtaining HRV genomic data and characterizing infections.


Recent advances in knowledge of zinc nutrition and human health.
Zinc deficiency increases the risk and severity of a variety of infections, restricts physical growth, and affects specific outcomes of pregnancy. Global recognition of the importance of zinc nutrition in public health has expanded dramatically in recent years, and more experience has accumulated on the design and implementation of zinc intervention programs. Therefore, the Steering Committee of the International Zinc Nutrition Consultative Group (IZiNCG) completed a second IZiNCG technical document that reexamines the latest information on the intervention strategies that have been developed to enhance zinc nutrition and control zinc deficiency. In particular, the document reviews the current evidence regarding preventive zinc supplementation and the role of zinc as adjunctive therapy for selected infections, zinc fortification, and dietary diversification or modification strategies, including the promotion and protection of breastfeeding and biofortification. The purposes of this introductory paper are to summarize new guidelines on the assessment of population zinc status, as recommended by the World Health Organization (WHO), the United Nations Children's Fund (UNICEF), the International Atomic Energy Agency (IAEA), and IZiNCG, and to provide an overview on several new advances in zinc metabolism. The following papers will then review the intervention strategies individually.


Evolution and epidemic spread of SARS-CoV-2 in Brazil.
Brazil currently has one of the fastest-growing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) epidemics in the world. Because of limited available data, assessments of the impact of nonpharmaceutical interventions (NPIs) on this virus spread remain challenging. Using a mobility-driven transmission model, we show that NPIs reduced the reproduction number from >3 to 1 to 1.6 in São Paulo and Rio de Janeiro. Sequencing of 427 new genomes and analysis of a geographically representative genomic dataset identified >100 international virus introductions in Brazil. We estimate that most (76%) of the Brazilian strains fell in three clades that were introduced from Europe between 22 February and 11 March 2020. During the early epidemic phase, we found that SARS-CoV-2 spread mostly locally and within state borders. After this period, despite sharp decreases in air travel, we estimated multiple exportations from large urban centers that coincided with a 25% increase in average traveled distances in national flights. This study sheds new light on the epidemic transmission and evolutionary trajectories of SARS-CoV-2 lineages in Brazil and provides evidence that current interventions remain insufficient to keep virus transmission under control in this country.


Dystroglycan in human fetal membranes decreases in later gestation and with spontaneous membrane rupture.
The transmembrane protein dystroglycan (DG) is known to anchor the cell membrane to the extracellular matrix, and is susceptible to cleavage by matrix metalloproteinases. This study tested the hypothesis that changes in DG abundance in fetal membranes (FM) occur late in gestation, with spontaneous rupture of membranes (SROM), and during labor. FM were collected from a prospective cohort consisting of four groups of patients (term labor, term unlabored, preterm labor, and preterm unlabored). FM were subjected to immunohistochemical staining using antibodies specific for α- and β-DG subunits, and staining intensity was graded by a blinded pathologist. α- and β-DG staining was significantly decreased at term and after SROM (p < 0.05), but not in the presence of labor. Decreased DG intensity was seen in FM of patients at term and with SROM, but no change was observed with labor.


Talent in autism: hyper-systemizing, hyper-attention to detail and sensory hypersensitivity.
We argue that hyper-systemizing predisposes individuals to show talent, and review evidence that hyper-systemizing is part of the cognitive style of people with autism spectrum conditions (ASC). We then clarify the hyper-systemizing theory, contrasting it to the weak central coherence (WCC) and executive dysfunction (ED) theories. The ED theory has difficulty explaining the existence of talent in ASC. While both hyper-systemizing and WCC theories postulate excellent attention to detail, by itself excellent attention to detail will not produce talent. By contrast, the hyper-systemizing theory argues that the excellent attention to detail is directed towards detecting 'if p, then q' rules (or [input-operation-output] reasoning). Such law-based pattern recognition systems can produce talent in systemizable domains. Finally, we argue that the excellent attention to detail in ASC is itself a consequence of sensory hypersensitivity. We review an experiment from our laboratory demonstrating sensory hypersensitivity detection thresholds in vision. We conclude that the origins of the association between autism and talent begin at the sensory level, include excellent attention to detail and end with hyper-systemizing.


Differential susceptibility to tetracycline, oxytetracycline and doxycycline of the calf pathogens Mannheimia haemolytica and Pasteurella multocida in three growth media.
For clinical isolates of bovine Mannheimia haemolytica and Pasteurella multocida, this study reports minimum inhibitory concentration (MIC) differences for tetracycline, oxytetracycline and doxycycline between cation-adjusted Mueller-Hinton broth (CAMHB), foetal bovine serum (FBS) and Roswell Park Memorial Institute (RPMI) medium. MICs were determined according to CLSI standards and additionally using five overlapping sets of twofold dilutions. Matrix effect: (a) free drug MICs and minimum bactericidal concentrations (MBC) for all drugs were significantly higher in FBS than in CAMHB for both pathogens (p < 0.001); (b) MICs and MBCs were higher for CAMHB and FBS compared to RPMI for P. multocida only. Net growth rate for P. multocida in CAMHB was significantly slower than in FBS and higher than in RPMI, correlating to MIC and MBC ranking. Drug effect: doxycycline MICs and MBCs were significantly lower (p < 0.001) in both CAMHB and FBS than tetracycline and oxytetracycline for both pathogens. Only for M. haemolytica were oxytetracycline MIC and MBC significantly lower than tetracycline, precluding the use of tetracycline to predict oxytetracycline susceptibility in this species. Determining potencies of tetracyclines in a physiological medium, such as FBS, is proposed, when the objective is correlation with pharmacokinetic data for dosage determination.


A pilot videoconference group stress management program in cancer survivors: lessons learned.
Cancer is a challenging experience and there is evidence that psychosocial interventions are effective at improving adjustment following treatment. At our cancer center, 14 cancer survivors (breast, prostate and blood cancers) completed a four-session cognitive-behavioral stress program. The first session was delivered at the survivor's local cancer center, where they were provided with a loaner tablet. The three subsequent sessions were delivered through group-based videoconference on the tablet. Session content was supplemented with a tailored ebook, designed specifically for this program. Participants provided feedback about the program as well as a standardized measure of perceived stress. Despite evidence that psychosocial programs are effective, there are significant barriers to dissemination, particularly for those residing in rural areas who do not live near academic medical centers where such programming is more readily available. Our experiences delivering a group-based videoconference program in cancer survivors are described, including positives and challenges associated with its design and implementation. Study participants enrolled from across four different US states, and the majority reported at least a 30-minute commute to their cancer center. This travel burden played a meaningful role in their desire to participate in our videoconference-based program. Although participants reported that session content was well suited to addressing stress management concerns, and session facilitators were able to effectively teach program techniques (eg progressive muscle relaxation, cognitive-reframing) and that the program was helpful overall, only modest improvements in perceived stress were seen. Participants noted challenges of the delivery including feeling disconnected from others, difficulty focusing, technical problems, and a desire for a longer program. Thus, although the novel delivery of a group-based, psychosocial program using tablet videoconference is feasible in a survivorship program, and desired by cancer survivors, key improvements must be made in future efforts. Our enthusiasm about the potential of telehealth must be tempered with the reality that such delivery can present challenges that interfere with the intervention implementation and efficacy. Facilitators must proactively address both the technological and interpersonal challenges associated with the use of group-based videoconference in order to improve its ability to positively impact cancer survivors. Many of these issues can be resolved prior to program launch, and require foresight and planning on the part of the program team.


From Mount Sinai to Mount Scopus: differences in the role and value of fine needle aspiration for evaluating thyroid nodules.
Fine needle aspiration is the main diagnostic tool used to assess thyroid nodules. To correlate FNA cytology results with surgical pathological findings in two teaching medical centers across the Atlantic. We retrospectively identified 484 patients at Hadassah Hebrew University Medical Center, Jerusalem and Mount Sinai Hospital, New York, by means of both preoperative FNA cytology and a final histopathological report. Results compared FNA diagnosis, histological findings and frozen section results (Mt. Sinai only). The sensitivity value of FNA at Hadassah was 83.0% compared with 79.1% at Mt. Sinai (NS). Specificity values were 86.6 vs. 98.5% (P < 0.05), negative predictive value 78.7 vs. 77.6% (NS) and positive predictive value 89.7 vs. 98.6% (P < 0.05), respectively. "Follicular lesion" was diagnosed on FNA in 33.1% of the patients at Hadassah and in 21.5% at Mt Sinai (P < 0.005) with a malignancy rate of 42.5 vs. 23.1% (P < 0.05), respectively. Frozen section was used in 190 patients at Mt. Sinai (78.5%) with sensitivity and specificity values of 72.3% and 100%. Frozen section results altered the planned operative course in only 6 patients (2.5%). Follicular carcinoma was diagnosed in 12 patients at Hadassah vs. 2 patients at Mt. Sinai (P < 0.05). The sensitivity of FNA at the two institutions was comparable. While malignancy on frozen section is highly specific, it should be used selectively for suspicious FNA results. Follicular lesions and the rate of malignancy in such lesions were more common at Hadassah, favoring a more aggressive surgical approach.


Efficacy of a novel combination of pneumatic energy and broadband light for the treatment of acne.
A novel photopneumatic platform (Isolaz, Pleasanton, CA), combining vacuum pressure with a broadband light source device has been designed to attack multiple targets for the effective treatment of acne. The objective of this study was to evaluate the safety and efficacy of photopneumatic technology for the treatment of mild to moderate acne vulgaris. Eleven subjects (7 women) aged 15 to 54 years with skin types 1 to 4 presented with mild to moderate facial acne (defined as 15 or more facial inflammatory or noninflammatory lesions) were recruited to the study. All subjects underwent 4 photopneumatic treatments at 3-week intervals with follow-up visits at 1 and 3 months. Inflammatory lesion counts continued to decrease for at least 3 months after the final treatment. At 3 months, reductions in lesion counts were significant for both inflammatory (P = .0137) and noninflammatory (P = .0383) lesions. Mean scores between visits consistently dropped sharply from their immediate posttreatment values for pain, erythema, and edema. Nine subjects (82%) were moderately satisfied to very satisfied with treatment. Results suggest that the photopneumatic device is a safe and effective modality for the treatment of mild to moderate inflammatory and comedonal acne vulgaris.


Human immunodeficiency virus reverse transcriptase. Substrate and inhibitor kinetics with thymidine 5'-triphosphate and 3'-azido-3'-deoxythymidine 5'-triphosphate.
3'-Azido-3'-deoxythymidine 5'-triphosphate (AZTTP) was an efficient substrate for the human immunodeficiency virus 1 reverse transcriptase. It was incorporated into both homopolymer and defined sequence DNA-primed RNA templates and DNA-primed DNA templates. The substrate and inhibitor kinetics of both AZTTP and dTTP were dependent on the template-primer and reaction conditions used. dTMP was incorporated into poly(rA).oligo(dT) and into a defined sequence DNA-primed RNA template (when the other three 2'-deoxynucleoside 5'-triphosphates were present) as a conventional substrate, with steady-state Km values of 5-10 microM. The results suggest that the reverse transcriptase was capable of processive DNA polymerization on these DNA-primed RNA templates. In contrast, in the absence of the other three 2'-deoxynucleoside 5'-triphosphates, the time course for incorporation of dTMP into the same defined sequence DNA-primed RNA template was biphasic. A burst of product formation was observed followed by a slow steady-state rate with a Km value of 0.082 microM. AZTMP incorporation into poly(rA).oligo(dT) and into the defined sequence DNA-primed RNA template produced similar biphasic time courses and steady-state Km values. These results were consistent with rate-limiting dissociation of the polymerase.template-primer complex after "forced" termination of polymerization. AZTMP and dTMP were both incorporated into the homopolymer DNA-primed DNA template, poly(dA).oligo(dT), and a defined sequence DNA-primed DNA template as conventional substrates. Their Km values were similar (2-10 microM). The absence of biphasic time courses suggested that dissociation of the DNA-primed DNA templates from the enzyme, after forced termination, was not rate-limiting. This was consistent with a more distributive mode of DNA polymerization. With the defined sequence template-primers and poly(dA).oligo(dT), Ki values for both dTTP and AZTTP were comparable to their Km values. Thus, AZTTP appeared to be a simple competitive substrate-inhibitor with respect to dTTP. AZTTP inhibition of dTMP incorporation into poly(rA).oligo(dT) was linear competitive at low concentrations (0-100 nM) of AZTTP (Ki = 35 nM) but became hyperbolic (decreasing potency) at concentrations of AZTTP above this range. A mechanism for this nonlinear inhibition is discussed.


Effects of object shape and visual feedback on hand configuration during grasping.
Normal subjects gradually preshape their hands during a grasping movement in order to conform the hand to the shape of a target object. The evolution of hand preshaping may depend on visual feedback about arm and hand position as well as on target shape and location at specific times during the movement. The present study manipulated object shape in order to produce differentiable patterns of finger placement along two orthogonal "dimensions" (flexion/extension and abduction/adduction), and manipulated the amount of available visual information during a grasp. Normal subjects were asked to reach to and grasp a set of objects presented in a randomized fashion at a fixed spatial location in three visual feedback conditions: Full Vision (both hand and target visible), Object Vision (only the object was visible but not the hand) and No Vision (vision of neither the hand nor the object during the movement). Flexion/extension angles of the metacarpophalangeal and proximal interphalangeal joints of the index, ring, middle and pinkie fingers as well as the abduction/adduction angles between the index-middle and middle-ring fingers were recorded. Kinematic analysis revealed that as visual feedback was reduced, movement duration increased and time to peak aperture of the hand decreased, in accord with previously reported studies. Analysis of the patterns of joint flexion/extension and abduction/adduction per object shape revealed that preshaping based on the abduction/adduction dimension occurred early during the reach for all visual feedback conditions (approximately 45% of normalized movement time). This early preshaping across visual feedback conditions suggests the existence of mechanisms involved in the selection of basic hand configurations. Furthermore, while configuration changes in the flexion/extension dimension resulting in well-defined hand configurations occurred earlier during the movement in the Object Vision and No Vision conditions (45%), those in the Full Vision condition were observed only after 75% of the movement, as the moving hand entered the central region of the visual field. The data indicate that there are at least two control mechanisms at work during hand preshaping, an early predictive phase during which grip selection is attained regardless of availability of visual feedback and a late responsive phase during which subjects may use visual feedback to optimize their grasp.


Excreted/secreted proteins from trypanosome procyclic strains.
Trypanosoma secretome was shown to be involved in parasite virulence and is suspected of interfering in parasite life-cycle steps such as establishment in the Glossina midgut, metacyclogenesis. Therefore, we attempted to identify the proteins secreted by procyclic strains of T. brucei gambiense and T. brucei brucei, responsible for human and animal trypanosomiasis, respectively. Using mass spectrometry, 427 and 483 nonredundant proteins were characterized in T. brucei brucei and T. brucei gambiense secretomes, respectively; 35% and 42% of the corresponding secretome proteins were specifically secreted by T. brucei brucei and T. brucei gambiense, respectively, while 279 proteins were common to both subspecies. The proteins were assigned to 12 functional classes. Special attention was paid to the most abundant proteases (14 families) because of their potential implication in the infection process and nutrient supply. The presence of proteins usually secreted via an exosome pathway suggests that this type of process is involved in trypanosome ESP secretion. The overall results provide leads for further research to develop novel tools for blocking trypanosome transmission.


Analysing self-regulatory behaviours in response to daily weighing: a think-aloud study with follow-up interviews.
Objective: To examine the extent to which people who are trying to lose weight naturally self-regulate in response to self-weighing and to identify barriers to self-regulation. Design/Main Outcome Measures: Twenty-four participants, who were overweight and trying to lose weight, recorded their thoughts during daily weighing for eight weeks. Semi-structured follow-up interviews assessed participant experiences. Qualitative analysis identified steps of the self-regulation process and barriers to self-regulation. Exploratory regression analysis assessed the relationship between the self-regulation steps and weight loss. Results: On 90% of 498 occasions, participants compared their weight measurement to an expectation or goal, and on 58% they reflected on previous behaviour. Action planning only occurred on 20% of occasions, and specific action planning was rare (6%). Only specific action planning significantly predicted weight loss (-2.1 kg per 1 SD increase in the predictor, 95% CI = -3.9, -0.3). Thematic analysis revealed that barriers to the interpretation of daily weight changes were difficulties in understanding day-to-day fluctuations, losing the overview of trends, forgetting to weigh, and forgetting previous measurements. Conclusion: Specific action planning can lead to weight loss, but is rare in a naturalistic setting. Barriers to self-regulation relate to the interpretation of weight changes.


Ultra-broadband EPR spectroscopy in field and frequency domains.
Electron paramagnetic resonance (EPR) is a powerful technique to investigate the electronic and magnetic properties of a wide range of materials. We present the first combined terahertz (THz) field and frequency domain electron paramagnetic resonance (HFEPR/FDMR) spectrometer designed to investigate the electronic structure and magnetic properties of molecular systems, thin films and solid state materials in a very broad frequency range of 85-1100 GHz. In this paper, we show high resolution frequency-field (Zeeman) maps (170-380 GHz by 0-15 T) recorded on two single-molecule magnets, [Mn2(saltmen)2(ReO4)2] and [Mn2(salpn)2(H2O)2](ClO4)2, which give direct access to the field-dependence of the energy level diagram. Furthermore, supression of standing waves in the described system and the sensitivity in field and frequency domain operations is evaluated and discussed.


Magnon Bose-Einstein condensation and spin superfluidity.
Bose-Einstein condensation (BEC) is a quantum phenomenon of formation of a collective quantum state in which a macroscopic number of particles occupy the lowest energy state and thus is governed by a single wavefunction. Here we highlight the BEC in a magnetic subsystem--the BEC of magnons, elementary magnetic excitations. The magnon BEC is manifested as the spontaneously emerging state of the precessing spins, in which all spins precess with the same frequency and phase even in an inhomogeneous magnetic field. The coherent spin precession was observed first in superfluid (3)He-B and this domain was called the homogeneously precessing domain (HPD). The main feature of the HPD is the induction decay signal, which ranges over many orders of magnitude longer than is prescribed by the inhomogeneity of magnetic field. This means that spins precess not with a local Larmor frequency, but coherently with a common frequency and phase. This BEC can also be created and stabilized by continuous NMR pumping. In this case the NMR frequency plays the role of a magnon chemical potential, which determines the density of the magnon condensate. The interference between two condensates has also been demonstrated. It was shown that HPD exhibits all the properties of spin superfluidity. The main property is the existence of a spin supercurrent. This spin supercurrent flows separately from the mass current. Transfer of magnetization by the spin supercurrent by a distance of more than 1 cm has been observed. Also related phenomena have been observed: the spin current Josephson effect; the phase-slip processes at the critical current; and the spin current vortex--a topological defect which is the analog of a quantized vortex in superfluids and of an Abrikosov vortex in superconductors; and so on. It is important to mention that the spin supercurrent is a magnetic phenomenon, which is not directly related to the mass superfluidity of (3)He: it is the consequence of a specific antiferromagnetic ordering in superfluid (3)He. Several different states of coherent precession have been observed in (3)He-B: the homogeneously precessing domain (HPD); a persistent signal formed by Q-balls at very low temperatures; coherent precession with fractional magnetization; and two new modes of coherent precession in compressed aerogel. In compressed aerogel the coherent precession has been also found in (3)He-A. We demonstrate that the coherent precession of magnetization is a true BEC of magnons, with the magnon interaction term in the Gross-Pitaevskii equation being provided by spin-orbit coupling which is different for different states of the magnon BEC.


An integrated semiconductor device enabling non-optical genome sequencing.
The seminal importance of DNA sequencing to the life sciences, biotechnology and medicine has driven the search for more scalable and lower-cost solutions. Here we describe a DNA sequencing technology in which scalable, low-cost semiconductor manufacturing techniques are used to make an integrated circuit able to directly perform non-optical DNA sequencing of genomes. Sequence data are obtained by directly sensing the ions produced by template-directed DNA polymerase synthesis using all-natural nucleotides on this massively parallel semiconductor-sensing device or ion chip. The ion chip contains ion-sensitive, field-effect transistor-based sensors in perfect register with 1.2 million wells, which provide confinement and allow parallel, simultaneous detection of independent sequencing reactions. Use of the most widely used technology for constructing integrated circuits, the complementary metal-oxide semiconductor (CMOS) process, allows for low-cost, large-scale production and scaling of the device to higher densities and larger array sizes. We show the performance of the system by sequencing three bacterial genomes, its robustness and scalability by producing ion chips with up to 10 times as many sensors and sequencing a human genome.


Atypical parasitic migration and necrotizing sacral myelitis due to Serratospiculoides amaculata in a prairie falcon (Falco mexicanus).
An adult, wild-caught, female prairie falcon (Falco mexicanus) was presented with the chief complaint of anorexia. Radiographic findings included increased densities within the air sacs, and coelomic endoscopy revealed numerous slender worms within the air sacs and on the serosal surfaces of the ovary, oviduct, liver, proventriculus, and ventriculus. The bird seemed to improve for a short period of time with antiparasitic therapy (ivermectin and fenbendazole) and supportive care. Twenty-one days after initial presentation, the bird became recumbent with increasing pelvic limb neurologic deficits and was euthanized. On histopathologic examination, mature nematodes and larvated eggs identified as Serratospiculoides amaculata were found within the subdural space of the distal thoracolumbar and synsacral spinal cord and within the coelomic cavity. This case suggests that S. amaculata can cause clinically significant lesions in its falconiform host with potentially fatal results.


Fingerprinting genomes using PCR with arbitrary primers.
Simple and reproducible fingerprints of complex genomes can be generated using single arbitrarily chosen primers and the polymerase chain reaction (PCR). No prior sequence information is required. The method, arbitrarily primed PCR (AP-PCR), involves two cycles of low stringency amplification followed by PCR at higher stringency. We show that strains can be distinguished by comparing polymorphisms in genomic fingerprints. The generality of the method is demonstrated by application to twenty four strains from five species of Staphylococcus, eleven strains of Streptococcus pyogenes and three varieties of Oryza sativa (rice).


Human aqueous humor phosphatase activity in cataract and glaucoma.
To investigate the presence and activity of protein phosphatase-2A (PPase2A), protein phosphatase-2C (PPase2C), and protein tyrosine phosphatases (PTPs) in the human aqueous humor (AH) of patients with primary open-angle glaucoma (POAG) and cataract and to study the correlation between these phosphatases and the redox state of the AH. Eighty-six cataract patients and 29 POAG patients who were scheduled for cataract surgery with or without glaucoma surgery were enrolled in the study. PPase2A, PPase2C, and PTPs levels in AH were measured by enzyme-linked immunosorbent assays, Western blot analyses, and spectral The redox state was measured by spectral and fluorescent methods. Phosphatase activity-positive results were significantly higher in AH samples from the POAG group (PP2A χ(2)(1) = 11.754, P < 0.01; PP2C χ(2)(1) = 8.754, P < 0.01; PTP χ(2)(1) = 11.073, P < 0.01). Western blot analysis revealed higher PP2C levels in the AH of glaucoma patients compared with PP2C levels in the AH of cataract patients (P = 0.012). Both oxidized/reduced glutathione ratios and superoxide dismutase levels in the AH were significantly higher in the glaucoma group than in the cataract group. Finally significant correlations were found between PP2A and PP2C, PP2A and PTP, and total antioxidant activity and PTP levels. There is a statistically significant difference between phosphatase levels in the AH of POAG patients and cataract patients. The phosphatase content of the AH represents tissue pathology, but their presence in the AH may be attributed to cell debris or to active signaling to other molecular events.


Balance asymmetry in Parkinson's disease and its contribution to freezing of gait.
Balance control (the ability to maintain an upright posture) is asymmetrically controlled in a proportion of patients with Parkinson's disease. Gait asymmetries have been linked to the pathophysiology of freezing of gait. We speculate that asymmetries in balance could contribute to freezing by a) hampering the unloading of the stepping leg and/or b) leading to a preferred stance leg during gait, which then results in asymmetric gait. To investigate this, we examined the relationship between balance control and weight-bearing asymmetries and freezing. We included 20 human patients with Parkinson (tested OFF medication; nine freezers) and nine healthy controls. Balance was perturbed in the sagittal plane, using continuous multi-sine perturbations, applied by a motion platform and by a force at the sacrum. Applying closed-loop system identification techniques, relating the body sway angle to the joint torques of each leg separately, determined the relative contribution of each ankle and hip joint to the total amount of joint torque. We also calculated weight-bearing asymmetries. We determined the 99-percent confidence interval of weight-bearing and balance-control asymmetry using the responses of the healthy controls. Freezers did not have larger asymmetries in weight bearing (p = 0.85) nor more asymmetrical balance control compared to non-freezers (p = 0.25). The healthy linear one-to-one relationship between weight bearing and balance control was significantly different for freezers and non-freezers (p = 0.01). Specifically, non-freezers had a significant relationship between weight bearing and balance control (p = 0.02), whereas this relation was not significant for freezers (p = 0.15). Balance control is asymmetrical in most patients (about 75 percent) with Parkinson's disease, but this asymmetry is not related to freezing. The relationship between weight bearing and balance control seems to be less pronounced in freezers, compared to healthy controls and non-freezers. However, this relationship should be investigated further in larger groups of patients.


StochPy: a comprehensive, user-friendly tool for simulating stochastic biological processes.
Single-cell and single-molecule measurements indicate the importance of stochastic phenomena in cell biology. Stochasticity creates spontaneous differences in the copy numbers of key macromolecules and the timing of reaction events between genetically-identical cells. Mathematical models are indispensable for the study of phenotypic stochasticity in cellular decision-making and cell survival. There is a demand for versatile, stochastic modeling environments with extensive, preprogrammed statistics functions and plotting capabilities that hide the mathematics from the novice users and offers low-level programming access to the experienced user. Here we present StochPy (Stochastic modeling in Python), which is a flexible software tool for stochastic simulation in cell biology. It provides various stochastic simulation algorithms, SBML support, analyses of the probability distributions of molecule copy numbers and event waiting times, analyses of stochastic time series, and a range of additional statistical functions and plotting facilities for stochastic simulations. We illustrate the functionality of StochPy with stochastic models of gene expression, cell division, and single-molecule enzyme kinetics. StochPy has been successfully tested against the SBML stochastic test suite, passing all tests. StochPy is a comprehensive software package for stochastic simulation of the molecular control networks of living cells. It allows novice and experienced users to study stochastic phenomena in cell biology. The integration with other Python software makes StochPy both a user-friendly and easily extendible simulation tool.


Role of pre-existing type 2 diabetes in colorectal cancer survival among older Americans: a SEER-Medicare population-based study 2002-2011.
Type 2 diabetes mellitus (diabetes) is a common comorbid condition among older adult colorectal cancer (CRC) patients, yet its effects on CRC mortality have not been adequately examined. This study aims to investigate the association between pre-existing diabetes, with and without complications, and CRC mortality. Medicare beneficiaries 67 years and older diagnosed with CRC between 2002 and 2011 were studied using the Surveillance, Epidemiology, and End Results (SEER)-Medicare datasets. Pre-existing diabetes was ascertained using validated algorithms. Cox proportional hazards models were used to compare all-cause and CRC-cause-specific death risk differences in relation to prior diabetes diagnosis and diabetes severity (with and without complications) with adjustment for relevant patient demographics and disease characteristics. Analyses included 93,710 CRC patients. Among the study population, 22,155 (24%) had diabetes prior to CRC diagnosis and 4% had diabetes-related complications (neuropathy, nephropathy, retinopathy, or peripheral circulatory disorders). All-cause CRC mortality was significantly higher among diabetic patients compared with non-diabetic patients (hazard ratio (HR) = 1.20; 95% confidence interval (CI) = 1.17-1.23). The results were more pronounced for diabetes with complications (HR = 1.47; 95% CI = 1.34-1.54). Diabetic patients with complications were 16% more likely to die of colorectal cancer compared with patients without diabetes (HR = 1.16; 95% CI = 1.08-1.25). Pre-existing diabetes contributes to poorer all-cause mortality among CRC patients and increased mortality from CRC among those with diabetes and complications. Opportunities exist to incorporate diabetes prevention and management interventions during CRC treatment phases among older adults.


Antimicrobial effect of spices and herbs on Vibrio parahaemolyticus.
The antimicrobial effects of spices and herbs from 18 plant species were examined on a foodborne pathogen, Vibrio parahaemolyticus, with the use of combinations of temperatures and nutrient levels. Basil, clove, garlic, horseradish, marjoram, oregano, rosemary, and thyme exhibited antibacterial activities at incubation of 30 degrees C, while with the exception of horseradish, the same spices and additional 7 species exhibited the activities at 5 degrees C. The lowest MIC (minimum inhibitory concentration) was 0.125% observed in clove and marjoram at 30 degrees C in a nutrient rich medium. Lowering of incubation temperature produced little effect on the MICs except for turmeric. The decreasing of the MIC in turmeric appeared to be basically attributed to the sensitivity of the bacterium to coldness. In nutrient poor medium, the lowest was 0.001 and 0.00025% in marjoram at 30 degrees C and at 5 degrees C, respectively. The sensitivity to several spices and herbs was similar among different clinical serotypes including the emerging strain O3:K6. These results suggest that the spices and herbs can be practical for protecting seafood from the risk of contamination by V. parahaemolyticus and used in hurdle technology with low temperature.


Structural comparison of typical and atypical E2 pestivirus glycoproteins.
Pestiviruses, within the family Flaviviridae, are economically important viruses of livestock. In recent years, new pestiviruses have been reported in domestic animals and non-cloven-hoofed animals. Among them, atypical porcine pestivirus (APPV) and Norway rat pestivirus (NRPV) have relatively little sequence conservation in their surface glycoprotein E2. Despite E2 being the main target for neutralizing antibodies and necessary for cell attachment and viral fusion, the mechanism of viral entry remains elusive. To gain further insights into the pestivirus E2 mechanism of action and to assess its diversity within the genus, we report X-ray structures of the pestivirus E2 proteins from APPV and NRPV. Despite the highly divergent structures, both are able to dimerize through their C-terminal domain and contain a solvent-exposed β-hairpin reported to be involved in host receptor binding. Functional analysis of this β-hairpin in the context of BVDV revealed its ability to rescue viral infectivity.


A systematic review protocol for reporting deficiencies within surgical case series.
Case series are an important and common study type in surgical literature. There is evidence that key data are excluded from published case series, and currently no reporting guideline exists for case series. There is, therefore, the potential to change practices and improve the reporting of case series. Reporting guidelines have been shown to be efficacious in raising the bar for reporting quality. We present our protocol for the first stage of guideline development--a systematic review of previously identified deficiencies in how surgical case series are reported. Electronic searches will be conducted on MEDLINE, EMBASE, Cochrane Methods Register, Science Citation Index and Conference Proceedings Citation Index, from the start of indexing until 5 November 2014. The electronic search strategy was developed with an information specialist. Two independent researchers will identify articles for inclusion, specifically those that describe reporting deficiencies within surgical case series. Data will be extracted to specifically focus on the deficiencies of reporting. These will be categorised according to their type, and other identified issues will also be presented. Data will be presented with descriptive statistics to determine frequently missing types of data, and the commonest reporting issues tabulated. The authors hope to disseminate the findings as widely as possible, irrespective of results, as these will add to the wider corpora of information on this subject. The systematic review will be published in a peer-reviewed journal and will be presented at a wide range of national and international conferences. Ultimately, this will inform a Delphi process for the development of a surgical case series reporting guideline. CRD42015016145.


A physiologically based clinical measure for spastic reflexes in spinal cord injury.
To test the validity of the Spinal Cord Assessment Tool for Spastic reflexes (SCATS), a clinical tool intended to rate spastic motor behavior after spinal cord injury (SCI). By using correlational analyses, the SCATS was validated using concurrent measurements of kinematics and electromyograms and traditional assessments of spasms and spastic hypertonia. Research laboratory (kinematics and electromyography) and outpatient medical clinic (traditional measures of spastic hypertonia). Eleven people with SCI were used for kinematic and electromyographic measurements. Seventeen people with SCI were used for comparison with other clinical scales. Not applicable. Main outcome measures Kinematic and surface electromyographic measurements of the tested lower extremity were used to quantify magnitude and/or duration of motor behaviors, and the Penn Spasm Frequency Scale (PSFS) and the Ashworth Scale were used to measure spasm frequency and resistance to joint movement for the hip flexors, knee flexors, and ankle plantarflexors, respectively. Concurrently, the SCATS was used to assess the clonus response to an imposed ankle dorsiflexion, the flexion response to a stimulus to the foot, and the knee extensor activity in response to an imposed leg extension. Each component of the SCATS was compared with the Ashworth Scale, the PSFS, and kinematic and electromyographic measurements by using the Spearman rank correlation test. Clonus, flexor spasm, and extensor spasm responses measured by using the SCATS correlated significantly with kinematic and electromyographic recordings (P<.01). Significant correlations were also observed between the SCATS extensor spasms and the Ashworth scores for hip and knee flexors and for ankle plantarflexors (rho=.98, .88, .61, respectively). Also, SCATS flexor spasms and SCATS clonus scores correlated significantly with some of the Ashworth scores. Only SCATS clonus scores correlated significantly with spasm frequency measures (rho=.59, P<.05). The SCATS produced a valid measure of 3 distinct types of spastic motor behaviors in SCI and may provide a complementary tool for measuring spastic hypertonia. Such a measure is valuable because current assessment tools do not differentiate between the different types of spastic motor behaviors that manifest after SCI. Distinguishing the 3 spastic reactions using an efficient and valid clinical tool may help guide management of spastic hypertonia in SCI.


Undercarboxylation of recombinant prothrombin revealed by analysis of gamma-carboxyglutamic acid using capillary electrophoresis and laser-induced fluorescence.
The gamma-carboxyglutamic acid (Gla) content of several variants of human prothrombin has been measured by using capillary electrophoresis and laser-induced fluorescence (CE-LIF). Both plasma-derived prothrombin and recombinant prothrombin contain ten residues of Gla per molecule of protein. In contrast, a variant of human prothrombin (containing the second kringle domain of bovine prothrombin) was separated into two populations that differed in their Gla content. Direct measurement of the Gla content showed an association with the presence or absence of the calcium-dependent conformational change that is required for prothombinase function. Thus, the CE-LIF assay is useful in determining the carboxylation status of recombinant proteins.


Distribution of technetium-99m-labeled multilamellar liposomes in patients with Hodgkin's disease.
The distribution of 99mTc-labeled multilamellar liposomes composed of dimyristoyl phosphatidylcholine (DMPC) and dimyristoyl phosphatidylglycerol (DMPG) at a molar ratio of 7:3, administered intravenously, was studied in ten patients with Hodgkin's disease (HD). The dose of lipid was 150 mg/m2 and the mean dose of radioactivity injected per patient was 8.1 mCi (range 6.7-9.8). Whole-body imaging techniques were used, and for each organ an uptake index was calculated as the percent photographic density (PD) relative to the PD of the liver. Results were compared to those in a group of six patients with other malignancies. Increased liposome uptake in several skeletal areas was observed in one patient with HD with diffuse bone involvement and in the bone marrow of two patients with HD with bone marrow involvement. No definite liposome uptake was observed in lymph nodes involved by HD or in tumor areas of patients with other malignancies. Patients with HD had a significantly higher uptake by bone marrow (23.8% compared with 10.2% at 4 hr p = 0.02), and lungs (59.6% compared with 25.0% at 4 hr, p = 0.01) than patients with other malignancies. Among patients with HD, the uptake by bone marrow and lungs were higher in those with constitutional symptoms (bone marrow at 4 hr 31.4% compared with 16.2%, p = 0.02; lungs at 4 hr 68.8% compared with 50.4%, p = 0.19) and with liver involvement (bone marrow at 4 hr 30.8% compared with 16.8%, p = 0.03; lungs at 4 hr 73.6% compared with 45.6%, p = 0.03). These results suggest that patients with HD have a different pattern of distribution of multilamellar liposomes which may be related to a combination of nonspecific stimulation of the reticuloendothelial system and tumor uptake. It does not appear that liposomal 99mTc is capable of adequately imaging HD for clinical diagnosis.


Charge-controlled microfluidic formation of lipid-based single- and multicompartment systems.
In this manuscript, we introduce a simple, off-the-shelf approach for the on-demand creation of giant unilamellar vesicles (GUVs) or multicompartment synthetic cell model systems in a high-throughput manner. To achieve this, we use microfluidics to encapsulate small unilamellar vesicles in block-copolymer surfactant-stabilized water-in-oil droplets. By tuning the charge of the inner droplet interface, adsorption of lipids can be either inhibited, leading to multicompartment systems, or induced, leading to the formation of droplet-stabilized GUVs. To control the charge density, we formed droplets using different molar ratios of an uncharged PEG-based fluorosurfactant and a negatively-charged PFPE carboxylic acid fluorosurfactant (Krytox). We systematically studied the transition from a multicompartment system to 3D-supported lipid bilayers as a function of lipid charge and Krytox concentration using confocal fluorescence microscopy, cryo-scanning electron microscopy and interfacial tension measurements. Moreover, we demonstrate a simple method to release GUVs from the surfactant shell and the oil phase into a physiological buffer - providing a remarkably high-yield approach for GUV formation. This widely applicable microfluidics-based technology will increase the scope of GUVs as adaptable cell-like compartments in bottom-up synthetic biology applications and beyond.


High-resolution analysis of genetic alterations in small bowel carcinoid tumors reveals areas of recurrent amplification and loss.
Carcinoid tumors of the small intestine are characterized by an indolent clinical course, secretion of neuropeptides, and resistance to standard cytotoxic chemotherapy. To evaluate the molecular events underlying carcinoid tumorigenesis, we used high-resolution arrays of single nucleotide polymorphisms to study chromosomal gains and losses in 24 primary and metastatic small bowel carcinoid tumors derived from 18 patients. Regions of gain or loss comprising whole chromosomes or large chromosomal regions constituted the most common class of anomalies. Loss of all or most of chromosome 18 was the commonest finding, evident in 11 of the 18 cases. Heterozygosity was also lost on chromosome arms 9p and 16q. The amplitude of observed gains was modest in comparison to those reported in some other tumor types. One focal region of recurrent gain on 14q mapped to the locus of the gene encoding the antiapoptotic protein DAD1, and immunohistochemical staining confirmed DAD1 protein expression in tumor samples. This detailed study of an uncommon neoplasm provides a basis to investigate putative oncogenes and tumor suppressor genes in intestinal carcinoid tumors.