[Formula: see text] Addressing the neurodevelopmental needs of children and adolescents with congenital heart disease: A review of the existing intervention literature.
Congenital heart disease (CHD) is among the most prevalent birth defects in the United States. Given that children with CHD are at risk for differences with development, learning, and psychosocial functioning, effective intervention becomes a central tenant of recommendations following neuropsychological consultation and evaluation. The primary focus of this review is to summarize available interventions for children and adolescents with CHD. The existing CHD literature has concentrated on early developmental services, psychopharmacological treatment, and need for academic supports. The literature is limited with regard to intervention studies that target cognitive deficits and psychosocial difficulties. To address this discrepancy, efficacious interventions that have been used to mitigate these concerns within other medical groups are also discussed in an effort to provide options for alternative recommendations and services. The current paper is intended to facilitate comprehensive care for cardiac patients by providing clinicians with a review of the available intervention literature, as well as potential interventions that may serve as supplemental strategies in the interim.


Targeting CDK1 and MEK/ERK Overcomes Apoptotic Resistance in BRAF-Mutant Human Colorectal Cancer.
The BRAFV600E mutation occurs in approximately 8% of human colorectal cancers and is associated with therapeutic resistance that is due, in part, to reactivation of MEK/ERK signaling cascade. Recently, pathway analysis identified cyclin-dependent kinase 1 (CDK1) upregulation in a subset of human BRAFV600E colorectal cancers. Therefore, it was determined whether CDK1 antagonism enhances the efficacy of MEK inhibition in BRAFV600E colorectal cancer cells. BRAFV600E colorectal cancer cell lines expressing CDK1 were sensitized to apoptosis upon siRNA knockdown or small-molecule inhibition with RO-3306 (CDK1 inhibitor) or dinaciclib (CDK1, 2, 5, 9 inhibitors). Combination of RO-3306 or dinaciclib with cobimetinib (MEK inhibitor) cooperatively enhanced apoptosis and reduced clonogenic survival versus monotherapy. Cells isogenic or ectopic for BRAFV600E displayed resistance to CDK1 inhibitors, as did cells with ectopic expression of constitutively active MEK CDK1 inhibitors induced a CASP8-dependent apoptosis shown by caspase-8 restoration in deficient NB7 cells that enhanced dinaciclib-induced CASP3 cleavage. CDK inhibitors suppressed pro-CASP8 phosphorylation at S387, as shown by drug withdrawal, which restored p-S387 and increased mitosis. In a colorectal cancer xenograft model, dinaciclib plus cobimetinib produced significantly greater tumor growth inhibition in association with a caspase-dependent apoptosis versus either drug alone. The Cancer Genome Atlas (TCGA) transcriptomic dataset revealed overexpression of CDK1 in human colorectal cancers versus normal colon. Together, these data establish CDK1 as a novel mediator of apoptosis resistance in BRAFV600E colorectal cancers whose combined targeting with a MEK/ERK inhibitor represents an effective therapeutic strategy.Implications: CDK1 is a novel mediator of apoptosis resistance in BRAFV600E colorectal cancers whose dual targeting with a MEK inhibitor may be therapeutically effective. Mol Cancer Res; 16(3); 378-89. ©2017 AACR.


Determinants of early initiation of breastfeeding in rural Tanzania.
Breastfeeding is widely known for its imperative contribution in improving maternal and newborn health outcomes. However, evidence regarding timing of initiation of breastfeeding is limited in Tanzania. This study examines the extent of and factors associated with early initiation of breastfeeding in three rural districts of Tanzania. Data were collected in 2011 in a cross-sectional survey of random households in Rufiji, Kilombero and Ulanga districts of Tanzania. From the survey, 889 women who had given birth within 2 years preceding the survey were analyzed. Both descriptive and inferential statistical analyses were conducted. Associations between the outcome variable and each of the independent variables were tested using chi-square. Logistic regression was used for multivariate analysis. Early initiation of breastfeeding (i.e. breastfeeding initiation within 1 h of birth) stood at 51 %. The odds of early initiation of breastfeeding was significantly 78 % lower following childbirth by caesarean section than vaginal birth (adjusted odds ratio (OR) = 0.22; 95 % confidence interval (CI) 0.14, 0.36). However, this was almost twice as high for women who gave birth in health facilities as for those who gave birth at home (OR = 1.75; 95 % CI 1.25, 2.45). Furthermore, maternal knowledge of newborn danger signs was negatively associated with early initiation of breastfeeding (moderate vs. high: OR = 1.73; 95 % CI 1.23, 2.42; low vs. high: OR = 2.06; 95 % CI 1.43, 2.96). The study found also that early initiation of breastfeeding was less likely in Rufiji compared to Kilombero (OR = 0.52; 95 % CI 0.31, 0.89), as well as among ever married than currently married women (OR = 0.46; 95 % CI 0.25, 0.87). To enhance early initiation of breastfeeding, using health facilities for childbirth must be emphasized and facilitated among women in rural Tanzania. Further, interventions to promote and enforce early initiation of breastfeeding should be devised especially for caesarean births. Women residing in rural locations and women who are not currently married should be specifically targeted with interventions aimed at enhancing early initiation of breastfeeding to ensure healthy outcomes for newborns.


The use of contrast enhanced ultrasound in carotid arterial disease.
Traditionally, stroke risk stratification has centred on the degree of internal carotid artery stenosis, and the presence of focal neurological symptoms. However, degree of stenosis alone is a relatively poor predictor of future stroke in asymptomatic patients; the Asymptomatic Carotid Surgery Trial highlighting the need to identify a subgroup of asymptomatics that may benefit from intervention. Attempting to define this subgroup has inspired imaging research to identify, in vivo, high-risk plaques. In addition to pre-operative risk stratification of carotid stenosis, contrast enhanced ultrasound (CEUS) may be employed in monitoring response to plaque-stabilising therapies. Unlike most contrast agents used for computed tomography and magnetic resonance imaging, microbubbles used in CEUS remain within the vascular space and can hence be used to study the vasculature. In addition to improving current carotid structural scans, CEUS has potential to add extra information on plaque characteristics. Furthermore, by targeting microbubbles to specific ligands expressed on vascular endothelium, CEUS may have the ability to probe plaque biology. This review describes the current carotid ultrasound examination and the need to improve it, rationale for imaging neovascularisation, use of CEUS to image neovascularisation, microbubbles in improving the structural imaging of plaque, potential problems with CEUS, and future directions.


Safety Assessment of Mentha mozaffarianii Essential Oil: Acute and Repeated Toxicity Studies.
Mentha mozaffarianii, an endemic species from the Labiatae family, is used in Iranian traditional medicine. This study evaluated the acute and repeated oral toxicity of the Mentha mozaffarianii essential oil (MMEO) in rats and mice. To assess the toxicity profile of the MMEO, we administered the essential oil to 48 rats and mice of both sexes by gavage in acute and repeated models. In acute toxicity, the animals were administered the MMEO (2000 mg/kg) and were monitored for 14 days. In the repeated toxicity, the MMEO was administered (100 mg/kg) daily for 4 weeks. On the 28th day, all the animals were scarified and blood and tissue samples were prepared. All the clinical, biochemical, and histopathological changes were assessed and compared with those in the controls. Statistical significance was determined by one- and two-way analyses of variance, followed by the Tukey test using GraphPad Prism 6. In the acute test, there was no mortality; therefore, the oral LD50 value determined in the mice and rats of both sexes was greater than 2000 mg/kg. In the repeated test, the animals received the MMEO and there was no mortality. In the biochemical analysis, there were significant increases in blood glucose, cholesterol, ALT, AST, ALP, and TSH in the female rats and also in BUN in the male rats. The histopathological studies revealed evidence of microscopic lesions in the liver, kidney, stomach, and small intestine tissues of the MMEO group. The results indicated that the acute toxicity of the MMEO in the mice and rats was of a low order and it revealed slight tissue damage to several organs when given subchronically at a dose of 100 mg/kg.


Psychometric performance of the English language six-item Caring Behaviours Inventory in an acute care context.
This study aimed to evaluate the psychometric performance of the six-item Caring Behaviours Inventory in a sample of Australian acute hospital inpatients. Caring is significant for nursing, and exploring the prevalence of staff-caring behaviours is imperative for high-quality acute care. There is a need for psychometrically sound scales that measures caring in acute care, without imposing extensive respondent burden. A cross-sectional survey design was used to distribute the six-item Caring Behaviours Inventory to an Australian sample of hospital inpatients (n = 210) in December 2012. Psychometric evaluation included item performance, construct validity and internal consistency reliability. The six-item Caring Behaviours Inventory had satisfactory psychometric performance as evidenced by normally distributed scores, a uni-dimensional structure explaining 65% of variance in data, a total Cronbach's α of 0·89 and corrected item-total correlations between 0·51-0·82. The six-item Caring Behaviours Inventory had satisfactory estimates of validity and reliability when tested in an Australian sample of acute hospital inpatients. The tool contributes to the literature by being a brief and nonburdensome alternative with seemingly strong psychometric properties to be used in future measures of caring in nursing. The six-item Caring Behaviours Inventory provides a psychometrically tested fundament for reflective clinical discussions on how nurse behaviours facilitate or impede patient experiences of caring. This can benefit quality development in clinical practice as being in tune with patient experiences and expectations is fundamental to high quality services and patient satisfaction.


Conductive Particles Enable Syntrophic Acetate Oxidation between Geobacter and Methanosarcina from Coastal Sediments.
Coastal sediments are rich in conductive particles, possibly affecting microbial processes for which acetate is a central intermediate. In the methanogenic zone, acetate is consumed by methanogens and/or syntrophic acetate-oxidizing (SAO) consortia. SAO consortia live under extreme thermodynamic pressure, and their survival depends on successful partnership. Here, we demonstrate that conductive particles enable the partnership between SAO bacteria (i.e., Geobacter spp.) and methanogens (Methanosarcina spp.) from the coastal sediments of the Bothnian Bay of the Baltic Sea. Baltic methanogenic sediments were rich in conductive minerals, had an apparent isotopic fractionation characteristic of CO2-reductive methanogenesis, and were inhabited by Geobacter and Methanosarcina As long as conductive particles were delivered, Geobacter and Methanosarcina persisted, whereas exclusion of conductive particles led to the extinction of Geobacter Baltic Geobacter did not establish a direct electric contact with Methanosarcina, necessitating conductive particles as electrical conduits. Within SAO consortia, Geobacter was an efficient [13C]acetate utilizer, accounting for 82% of the assimilation and 27% of the breakdown of acetate. Geobacter benefits from the association with the methanogen, because in the absence of an electron acceptor it can use Methanosarcina as a terminal electron sink. Consequently, inhibition of methanogenesis constrained the SAO activity of Geobacter as well. A potential benefit for Methanosarcina partnering with Geobacter is that together they competitively exclude acetoclastic methanogens like Methanothrix from an environment rich in conductive particles. Conductive particle-mediated SAO could explain the abundance of acetate oxidizers like Geobacter in the methanogenic zone of sediments where no electron acceptors other than CO2 are available.IMPORTANCE Acetate-oxidizing bacteria are known to thrive in mutualistic consortia in which H2 or formate is shuttled to a methane-producing Archaea partner. Here, we discovered that such bacteria could instead transfer electrons via conductive minerals. Mineral SAO (syntrophic acetate oxidation) could be a vital pathway for CO2-reductive methanogenesis in the environment, especially in sediments rich in conductive minerals. Mineral-facilitated SAO is therefore of potential importance for both iron and methane cycles in sediments and soils. Additionally, our observations imply that agricultural runoff or amendments with conductive chars could trigger a significant increase in methane emissions.


Current research in transcultural psychiatry in the Nordic countries.
This article discusses major themes in recent transcultural psychiatric research in the Nordic countries from 2008 to 2011: (a) epidemiological studies of migration, (b) indigenous populations, and (c) quality of psychiatric care for migrants. Over the past several decades, the populations of the Nordic countries, Denmark, Finland, Norway, and Sweden, which were relatively homogeneous, have become increasingly culturally diverse. Many migrants to Nordic countries have been exposed to extreme stress, such as threats of death and/or torture and other severe social adversities before, during, and after migration, with potential effects on their physical, mental, social, and spiritual health. Growing interest in transcultural issues is reflected in the level of scientific research and clinical activity in the field by Nordic physicians, psychologists, social scientists, demographers, medical anthropologists, as well as other clinicians and policy planners. Research includes work with migrants and indigenous minorities in the Nordic countries, as well as comparisons with mental health in postconflict countries. We conclude by suggesting future directions for transcultural psychiatry research and providing guidelines for the education and training of future clinicians in the Nordic countries.


Community-Based Upper Gastrointestinal Cancer Screening in a Randomized Controlled Trial: Baseline Results in a Non-high-incidence Area.
A cluster-randomized controlled trial (RCT) was conducted to evaluate to the effectiveness of reducing mortality of upper gastrointestinal cancer (UGC) and feasibility of screening through a questionnaire combined with endoscopy in non-high-incidence urban areas in China. The trial design, recruitment performance, and preliminary results from baseline endoscopy are reported. Seventy-five communities in two urban cities with a non-high-incidence of UGC were randomized to a screening endoscopy arm (n = 38) or a control arm (n = 37). In the screening arm, individuals at high risk of UGC underwent endoscopic screening. The primary outcome was the UGC mortality, and secondary outcomes included the UGC detection rate, incidence rate, survival rate, and clinical stage at the time of diagnosis. A total of 10,416 and 9,565 individuals were recruited into the screening and control arms, respectively. The participation rate was 74.3%. In the screening arm, 5,242 individuals (50.3%) were estimated to be high-risk. Among them, 2,388 (45.6%) underwent endoscopic screening. Age and household income were associated with undergoing endoscopy. Three early esophageal cancer (0.13%), one gastric cancer (0.04%), 29 precancerous esophageal lesions (1.21%), and 53 precancerous gastric lesions (2.22%) were detected. Age, sex, a family history of cancer, intake of meat-egg-milk frequently, superficial gastritis, and clinical symptoms of gastric cancer were associated with the presence of precancerous lesions. The detection rate was low using endoscopic screening in non-high-incidence area given the relatively low compliance rate. These findings provide a reference for designing effective community-based UGC screening strategies in non-high-incidence urban areas.


Comparison of the nutrient content of children's menu items at US restaurant chains, 2010-2014.
To determine changes in the nutritional content of children's menu items at U.S. restaurant chains between 2010 and 2014. The sample consisted of 13 sit down and 16 fast-food restaurant chains ranked within the top 50 US chains in 2009. Nutritional information was accessed in June-July 2010 and 2014. Descriptive statistics were calculated for nutrient content of main dishes and side dishes, as well as for those items that were added, removed, or unchanged during the study period. Nutrient content of main dishes did not change significantly between 2010 and 2014. Approximately one-third of main dishes at fast-food restaurant chains and half of main dishes at sit down restaurant chains exceeded the 2010 Dietary Guidelines for Americans recommended levels for sodium, fat, and saturated fat in 2014. Improvements in nutrient content were observed for side dishes. At sit down restaurant chains, added side dishes contained over 50% less calories, fat, saturated fat, and sodium, and were more likely to contain fruits/vegetables compared to removed sides (p < 0.05 for all comparisons). Added side dishes at fast-food restaurant chains contained less saturated fat (p < 0.05). The majority of menu items, especially main dishes, available to children still contain high amounts of calories, fat, saturated fat, and sodium. Efforts must be made by the restaurant industry and policy makers to improve the nutritional content of children's menu items at restaurant chains to align with the Dietary Guidelines for Americans. Additional efforts are necessary to help parents and children make informed choices when ordering at restaurant chains.


[Clinical studies of pediatric malabsorption syndromes].
Multiple cases with various types of pediatric malabsorption syndromes were evaluated. The clinical manifestations, laboratory findings, pathophysiology, and histopathological descriptions of each patient were analyzed in an effort to clear the pathogenesis of the malabsorption syndromes and the treatments were undertaken. The cases studied, included one patient with cystic fibrosis, two with lactose intolerance with lactosuria (Durand type), one with primary intestinal lymphangiectasia, two with familial hypobetalipoproteinemia, one with Hartnup disease, one with congenital chroride diarrhea, one with acrodermatitis enteropathica, one with intestinal nodular lymphoid hyperplasia (NLH), five with intractable diarrhea of early infancy and four with glycogenosis type Ia. Each case description and outcome is described below: 1. A 15-year-old Japanese boy with cystic fibrosis presented with severe symptoms, including pancreatic insufficiency, bronchiectasis, pneumothorax and hemoptysis. His prognosis was poor. Analysis of the CFTR genes of this patient revealed a homozygous large deletion from intron 16 to 17b. 2. In the sibling case of Durand type lactose intolerance, the subjects'disaccaridase activity of the small bowel, including lactase, were within normal limits. The results of per oral and per intraduodenal lactose tolerance tests confirmed lactosuria in both. These observations suggested, not only an abnormal gastric condition, but also duodenal and intestinal mucosal abnormal permeability of lactose. 3. In the case of primary intestinal lymphangiectasia, the subject had a lymphedematous right arm and hand, a grossly coarsened mucosal pattern of the upper gastrointestinal tract (identified via radiologic examination) and the presence of lymphangiectasia (confirmed via duodenal mucosal biopsy). The major laboratory findings were hypoalbuminemia, decreased immunoglobulin levels and lymphopenia resulting from loss of lymph fluid and protein into the gastro-intestinal tract. 4. In two cases of heterozygous familial hypobetalipoproteinemia, serum total cholesterol and betalipoprotein levels were very low. The subjects presented with symptoms and signs of acanthocytosis and fat malabsorption. Further, one subject had neurological abnormalities such as mental retardation and severe convulsions. Treatment with MCT formula diet corrected the lipid malabsorption. 5. A 5-year-old girl presented with pellagra-like rashes, mental retardation and cerebellar ataxia. An oral tryptophan (Trp) and dipeptide (Trp-Phe) loading test were conducted and the renal clearance of amino acids was also evaluated in this patient and in controls. Following the oral Trp loading test, plasma levels of Trp indicated a lower peak in the case, reaching a maximum at 60 minutes. On the other hand, the oral dipeptide (Trp-Phe) loading test in the Hartnup patient showed the peak Trp plasma level was the same as the control subjects. The renal clearance of neutral amino acids in this case increased to levels 5 to 35 times normal. 6. In the case of congenital chloride diarrhea, the subject had secondary lactose intolerance, dehydration, hyponatremia, hypokalemia, hypochloremia, hyperreninemia and metabolic alkalosis. The chloride content of her fecal fluid was very high. The concentrations were 89-103 mEq/l. In contrast, her urine was chloride-free. The subject's growth and development improved after treatment with lactose free formura and oral replacement of the fecal loses of water, NaCl and KCl. Unfortunately, the patient died of a small bowel intussusception. The kidney histopathological finding was juxtaglomerular hyperplasia by a necropsy. 7. In the case of acrodermatitis enteropathica, the subject had characteristic skin lesions, low serum zinc levels and ALPase activity. An oral ZnSO4 loading test and intestinal mucosal histology by a peroral biopsy were conducted. The serum zinc peak level was 2 hours after the oral ZnSO4 loading test. Infant formula alone could not maintain normal serum zinc ranges. Light microscopic studies of the intestinal villous architecture showed a normal pattern. However, ultrastructual examination of several epithelial cells revealed numerous intracellular vesicles. After zinc therapy, these changes were decreased. The lesions were postulated as the secondary result of zinc deficiency. 8. A 12-year-old girl presented with hypogammaglobulinemia, recurrent infections, chronic diarrhea and intestinal NLH. A barium meal and follow-through examination showed multiple nodules throughout the stomach and intestine. The nodules, all uniform in size, were 2 mm diameter. The barium enema did not show NLH in the colon. Mucosal biopsy of the stomach and jejunum revealed the typical histology of NLH in the lamina propria. Also, achlorhydria was present in this patient and her serum gastrin levels were very high; 315-775 pg/ml. 9. In 4 cases of intractable diarrhea in early infancy (by Avery G B), a jejunal biopsy showed shortening villi and nonspecific enterocolitis. Some patients were found with only low lactase or low lactase and sucrase levels. An electron microscope analysis of the small bowel in 2 cases showed alterations: increased pinocytosis in microvillus membranes and lysosomes by endocytosis of undigested macromolecular substances. I postulated that the stated evidence was causative of this clinical profile. 10. I frequently observed diarrhea as a clinical manifestation in glycogenosis type Ia and lipid malabsorption in one case. The light and electron photomicrographs showed intestinal absorption cells with the glycogen deposits in the inferior devision of nuclei.


Integrative analysis of mRNA and microRNA expression profiles in laryngeal squamous cell carcinoma.
Larynx cancer is a therapeutically challenging disease. Rapidly evolving experimentally validated data have significantly improved our understanding of the complex role of numerous RNA, DNA, and proteins that play a role in the development and progression of cancer. Based on the insights from approximately two decades of research, it seems clear that microRNAs (miRNAs) have revolutionized our concepts related to the main role of noncoding RNAs in different cancers' progression, development, and metastasis. Mechanistically, miRNAs have been reported to regulate different RNAs and finally protein-coding genes. The expression profiling of miRNAs and messenger RNA (mRNAs) was conducted for a deeper analysis of the miRNAs and mRNAs which play an essential role in larynx cancer. Downregulation or upregulation over twofolds in the miRNAs was considered to be significant, and that of sixfolds or below was considered to be significant for the mRNAs. In accordance with this approach, the expression levels of 43 miRNAs were increased in this study, whereas the expression levels of 129 were decreased. Accordingly, all the genomic expression studies provided evidence of upregulation of 97 genes, whereas 128 genes were found to be downregulated. Among these miRNAs, hsa-miR-20a-3p and hsa-miR-1972 were noted to be important in the etiology of larynx cancer.


Differences between work and leisure in temporal patterns of objectively measured physical activity among blue-collar workers.
Leisure time physical activity (LTPA) is generally associated with favorable cardiovascular health outcomes, while occupational physical activity (OPA) shows less clear, or even opposite, cardiovascular effects. This apparent paradox is not sufficiently understood, but differences in temporal patterns of OPA and LTPA have been suggested as one explanation. Our aim was to investigate the extent to which work and leisure (non-occupational time) differ in temporal activity patterns among blue-collar workers, and to assess the modification of these patterns by age and gender. This study was conducted on a cross-sectional sample of male (n = 108) and female (n = 83) blue-collar workers, aged between 21 and 65 years. Physical activity and sedentary behavior were assessed using accelerometers (Actigraph GT3X+) worn on the thigh and trunk for four consecutive days. Temporal patterns of OPA and LTPA were retrieved using Exposure Variation Analysis (EVA), and expressed in terms of percentage of work and leisure time spent in uninterrupted periods of different durations (<1 min, 1-5 min, 5-10 min, 10-30 min, 30-60 min and > 60 min) of sitting, standing, and walking. Repeated measures ANOVA and linear regression analyses were used to test a) possible differences between OPA and LTPA in selected EVA derivatives, and b) the modification of these differences by age and gender. OPA showed a larger percentage time walking in brief (<5 min) periods [mean (SD): 33.4 % (12.2)], and less time in prolonged (>30 min) sitting [7.0 % (9.3)] than LTPA [walking 15.4 % (5.0); sitting 31.9 % (15.3)], even after adjustment for the difference between work and leisure in total time spent in each activity type. These marked differences in the temporal pattern of OPA and LTPA were modified by gender, but not age. We found that the temporal patterns of OPA and LTPA among blue-collar workers were markedly different even after adjustment for total physical activity time, and that this difference was modified by gender. We recommend using EVA derivatives in future studies striving to disentangle the apparent paradoxical cardiovascular effect of physical activity at work and during leisure.


Detection and quantitation of 5-HT1A and 5-HT2A receptor mRNAs in human hippocampus using a reverse transcriptase-polymerase chain reaction (RT-PCR) technique and their correlation with binding site densities and age.
The presence and abundance of 5-HT1A and 5-HT2A receptor mRNAs in post mortem human hippocampus was investigated using a novel quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) technique using cyclophilin mRNA as an internal standard. 5-HT1A and 5-HT2A receptor mRNAs were each co-amplified with varying dilutions of cyclophilin primers, and their abundance expressed as a ratio of cyclophilin mRNA. Using this technique in combination with quantitative autoradiography we have investigated the effect of aging on hippocampal 5-HT1A and 5-HT2A receptor mRNA abundance and binding site densities. There was a significant negative correlation between hippocampal 5-HT1A receptor binding site densities and age and a similar trend for 5-HT1A receptor mRNA abundance. Neither 5-HT2A receptor binding site densities nor mRNA abundance were affected by age. Both 5-HT1A and 5-HT2A receptor binding site densities in individual subjects correlated significantly with abundance of their encoding mRNA. This study demonstrates that 5-HT1A and 5-HT2A receptor mRNAs can be measured in small samples of human brain. Combining studies of mRNA with those directed at binding sites will help reveal mechanisms underlying changes in expression of these receptors in various neuropsychiatric disorders.


Comparison of dentin bond durability of a universal adhesive and two etch-and-rinse adhesive systems.
This study aimed to compare dentin bond durability under different degradation conditions between two etch-and-rinse (ER) systems and a universal adhesive in ER mode. This study used a universal adhesive [Scotchbond Universal (SU)], a three-step ER adhesive [Scotchbond Multi-Purpose Plus (SM)], and a two-step ER adhesive [Single Bond Plus (SB)]. A phosphoric acid-etching agent was applied to bovine dentin prior to the application of either a primer or the adhesive. After acid etching, bonding procedures were conducted. The specimens were divided into three group classes: (1) subjected to 10,000, 30,000, or 50,000 thermal cycles (TC); (2) stored in distilled water at 37 °C for 6 months or 1 year (WS); and (3) stored in distilled water for 24 h (baseline). Shear bond strength (SBS) tests were conducted. SB showed a higher baseline SBS than the other adhesives. Defining the baseline SBS value for each adhesive system as 100%, TC groups ranged from 56.1 to 70.3% for SM, from 98.4 to 103.7% for SB, and from 120.3 to 126.7% for SU. WS groups ranged from 66.2 to 71.4% for SM, from 98.1 to 103.3% for SB, and from 102.5 to 118.1% for SU. Although SB showed relatively stable dentin bond performance under all degradation conditions, SM showed decreased dentin SBS with prolonged degradation. SU did not show any significant decrease in SBS from the baseline under any degradation condition. The universal adhesive showed comparable adhesive performance with the two-step ER adhesive.


Altered hippocampal microstructure and function in children who experienced Hurricane Irma.
Hurricane Irma was the most powerful Atlantic hurricane in recorded history, displacing 6 million and killing over 120 people in the state of Florida alone. Unpredictable disasters like Irma are associated with poor cognitive and health outcomes that can disproportionately impact children. This study examined the effects of Hurricane Irma on the hippocampus and memory processes previously related to unpredictable stress. We used an innovative application of an advanced diffusion-weighted imaging technique, restriction spectrum imaging (RSI), to characterize hippocampal microstructure (i.e., cell density) in 9- to 10-year-old children who were exposed to Hurricane Irma relative to a non-exposed control group (i.e., assessed the year before Hurricane Irma). We tested the hypotheses that the experience of Hurricane Irma would be associated with decreases in: (a) hippocampal cellularity (e.g., neurogenesis), based on known associations between unpredictable stress and hippocampal alterations; and (b) hippocampal-related memory function as indexed by delayed recall. We show an association between decreased hippocampal cellularity and delayed recall memory in children who experienced Hurricane Irma relative to those who did not. These findings suggest an important role of RSI for assessing subtle microstructural changes related to functionally significant changes in the developing brain in response to environmental events.


The use of monoclonal antibodies to analyze the structure of Clostridium botulinum type E derivative toxin.
Six monoclonal antibodies against Clostridium botulinum type E derivative toxin were prepared. Three of the five binding to the heavy chain neutralized the derivative toxin; the other one binding to the light chain did not. Immunoblotting analysis with the monoclonal antibodies showed that the fragment obtained by tryptic digestion consisted of the light chain and part of the heavy chain (H-1 fragment) linked together by a disulfide bond(s) and that the antigenic determinants common between type E and F derivative toxins were located on both the heavy and light chains. The fragment induced by chymotrypsin treatment, like the tryptic fragment, bound to four monoclonal antibodies. The mild tryptic treatment and reduction resulted in separation of the chymotryptic fragment into two smaller fragments corresponding to the light chain and H-1 fragment. These results indicate that H-1 fragment contains the amino-terminal portion of the heavy chain. The monoclonal antibody neutralizing the toxin and probably recognizing the epitope on the carboxyl-terminal portion (H-2 fragment) of the heavy chain effectively competed for binding of 125I-labeled derivative toxin to synaptosomes. Of the two monoclonal antibodies neutralizing the toxin and recognizing the epitopes on H-1 fragment, one partially inhibited binding, but the other did not. This suggests that the binding of 125I-labeled derivative toxin depends mainly on the carboxyl-terminal region of the heavy chain and that interference with binding is not the only means of toxin neutralization.


Visual perspective taking and laterality decisions: Problems and possible solutions.
Perspective taking plays an important role in different areas of psychological and neuroscientific research. Visual perspective taking is an especially prominent approach generally using one of two experimental tasks: in the own-body-transformation task observers are asked to judge the laterality of a salient feature of a human figure (e.g., is the glove on the left or right hand?) from the figure's perspective. In the avatar-in-scene task they decide about the laterality of objects in a scene (e.g., is the flower on the left or right?) from the avatar's point of view. Increases in latencies and/or errors are interpreted as originating from additional cognitive processes predominately described as observer-based perspective transformations. A closer look reveals that such an account is disputable on grounds related to the use of laterality judgments. Other transformation accounts, i.e., object or array transformations, as well as non-transformational accounts, i.e., extra processing due to spatial conflicts, have not been adequately considered, tested, or ruled out by existing research. Our review examines visual perspective tasks in detail, identifies problems and makes recommendations for future research.


Evaluating the Effectiveness and Safety of the Electroencephalogram-Based Brain-Machine Interface Rehabilitation System for Patients With Severe Hemiparetic Stroke: Protocol for a Randomized Controlled Trial (BEST-BRAIN Trial).
We developed a brain-machine interface (BMI) system for poststroke patients with severe hemiplegia to detect event-related desynchronization (ERD) on scalp electroencephalogram (EEG) and to operate a motor-driven hand orthosis combined with neuromuscular electrical stimulation. ERD arises when the excitability of the ipsi-lesional sensorimotor cortex increases. The aim of this study was to evaluate our hypothesis that motor training using this BMI system could improve severe hemiparesis that is resistant to improvement by conventional rehabilitation. We, therefore, planned and implemented a randomized controlled clinical trial (RCT) to evaluate the effectiveness and safety of intensive rehabilitation using the BMI system. We conducted a single blind, multicenter RCT and recruited chronic poststroke patients with severe hemiparesis more than 90 days after onset (N=40). Participants were randomly allocated to the BMI group (n=20) or the control group (n=20). Patients in the BMI group repeated 10-second motor attempts to operate EEG-BMI 40 min every day followed by 40 min of conventional occupational therapy. The interventions were repeated 10 times in 2 weeks. Control participants performed a simple motor imagery without servo-action of the orthosis, and electrostimulation was given for 10 seconds for 40 min, similar to the BMI intervention. Overall, 40 min of conventional occupational therapy was also given every day after the control intervention, which was also repeated 10 times in 2 weeks. Motor functions and electrophysiological phenotypes of the paretic hands were characterized before (baseline), immediately after (post), and 4 weeks after (follow-up) the intervention. Improvement in the upper extremity score of the Fugl-Meyer assessment between baseline and follow-up was the main outcome of this study. Recruitment started in March 2017 and ended in July 2018. This trial is currently in the data correcting phase. This RCT is expected to be completed by October 31, 2018. No widely accepted intervention has been established to improve finger function of chronic poststroke patients with severe hemiparesis. The results of this study will provide clinical data for regulatory approval and novel, important understanding of the role of sensory-motor feedback based on BMI to induce neural plasticity and motor recovery. UMIN Clinical Trials Registry UMIN000026372; https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi? recptno=R000030299 (Archived by WebCite at http://www.webcitation.org/743zBJj3D). DERR1-10.2196/12339.


Onset and maintenance of psychiatric disorders after serious accidents.
The purpose of this study was to prospectively investigate the onset, course, and remission of psychiatric disorders in the first 6 months after a serious accident for consecutive patients in a hospital emergency department. Participants were 58 patients aged 18-65 who were assessed shortly after attending a hospital emergency department and were followed up 6 months afterwards. Patients were interviewed with regard to past and current psychiatric history using different instruments (e.g. SCID for DSM-IV). Prior to their accidents, 35% of all subjects had experienced one or more psychiatric disorders (lifetime prevalence). Shortly after the accident, the incidence of Acute Stress Disorder (7%), subsyndromal Acute Stress Disorder (12%), and adjustment disorder (1.5%) was increased as a reaction to the accident. At this time, 29% of all patients suffered from an acute psychiatric disorder. Six-months after the accident, 10% of the subjects met criteria for Major Depression, 6% for PTSD, 4% for subsyndromal PTSD, and 1.5% for Specific Phobia as newly developed disorders. The course of the psychiatric disorders shows that those patients who met criteria for any psychiatric diagnosis shortly after the accident ran a much higher risk for developing new or comorbid psychiatric disorders in the future.


Current findings, future trends, and unsolved problems in studies of medicinal mushrooms.
The target of the present review is to draw attention to many critically important unsolved problems in the future development of medicinal mushroom science in the twenty-first century. Special attention is paid to mushroom polysaccharides. Many, if not all, higher Basidiomycetes mushrooms contain biologically active polysaccharides in fruit bodies, cultured mycelium, and cultured broth. The data on mushroom polysaccharides are summarized for approximately 700 species of higher Hetero- and Homobasidiomycetes. The chemical structure of polysaccharides and its connection to antitumor activity, including possible ways of chemical modification, experimental testing and clinical use of antitumor or immunostimulating polysaccharides, and possible mechanisms of their biological action, are discussed. Numerous bioactive polysaccharides or polysaccharide-protein complexes from medicinal mushrooms are described that appear to enhance innate and cell-mediated immune responses and exhibit antitumor activities in animals and humans. Stimulation of host immune defense systems by bioactive polymers from medicinal mushrooms has significant effects on the maturation, differentiation, and proliferation of many kinds of immune cells in the host. Many of these mushroom polymers were reported previously to have immunotherapeutic properties by facilitating growth inhibition and destruction of tumor cells. While the mechanism of their antitumor actions is still not completely understood, stimulation and modulation of key host immune responses by these mushroom polymers appears central. Particularly and most importantly for modern medicine are polysaccharides with antitumor and immunostimulating properties. Several of the mushroom polysaccharide compounds have proceeded through phases I, II, and III clinical trials and are used extensively and successfully in Asia to treat various cancers and other diseases. A total of 126 medicinal functions are thought to be produced by medicinal mushrooms and fungi including antitumor, immunomodulating, antioxidant, radical scavenging, cardiovascular, antihypercholesterolemia, antiviral, antibacterial, antiparasitic, antifungal, detoxification, hepatoprotective, and antidiabetic effects.


Adolescents' reports of physical violence by peers in residential care settings: an ecological examination.
Physical victimization by peers was examined among 1,324 Jewish and Arab adolescents, aged 11 to 19, residing in 32 residential care settings (RCS) for children at-risk in Israel. Hierarchical Linear Modeling (HLM) was used to examine the relationships between physical victimization and adolescents' characteristics (age, gender, self-efficacy, adjustment difficulties, maltreatment by staff, and perceived social climate) as well as institution-level characteristics (care setting type, size, structure, and ethnic affiliation). For this study, we define physical violence as being grabbed, shoved, kicked, punched, hit with a hand, or hit with an object. Over 50% (56%) of the adolescents surveyed reported having experienced at least one form of physical violence by peers. Boys and younger adolescents were more likely to be victimized than girls and older adolescents. The results show that adolescents with adjustment difficulties or low social self-efficacy, and adolescents who perceive an institution's staff as strict and/or had experienced maltreatment by staff, are vulnerable groups for peer victimization. Lower levels of victimization were found in RCS with a familial element than in traditional group settings. Institutions with high concentrations of young people with adjustment difficulties and violent staff behaviors had higher levels of violence among residents. Applying an ecological perspective to an investigation of peer victimization in RCS enables the identification of risk factors at adolescent and institution levels. This type of examination has implications for child welfare practice and policy that can help in the development of prevention and intervention methods designed to tackle the involvement in violence of youth in care.


Rubella serosurveys at three Aravind Eye Hospitals in Tamil Nadu, India.
To determine the susceptibility of female eye hospital staff to rubella infection and the potential risk for hospital-based rubella outbreaks. A prospective cohort study on the seroprevalence of rubella IgG antibodies was conducted at three large eye hospitals in Coimbatore, Madurai and Tirunelveli, Tamil Nadu, India, where young children with eye abnormalities attributable to congenital rubella are treated. A total of 1000 female hospital employees aged 18-40 years agreed to participate and gave written informed consent. The proportions of rubella-seronegative women were: 11.7% at Coimbatore, with a 95% confidence interval (CI) of 8.1-16.5; 15% at Madurai (95% CI = 12.3-18.1), and 20.8 at Tirunelveli (95% CI = 14.7-28.6). For the entire cohort the proportion seronegative was significantly higher among married women (21.5%) than among single women (14.0%) (P = 0.02). Rates of seronegativity were highest among physicians and lowest among housekeepers. All 150 seronegative women in the study sample accepted a dose of rubella vaccine. These are the first rubella serosurveys to have been reported from eye hospitals in any country. The relatively high rate of susceptibility indicated a risk of a rubella outbreak, and this was reduced by vaccinating all seronegative women. A policy has been established at all three hospitals for the provision of rubella vaccine to new employees. Other hospitals, especially eye hospitals and hospitals in countries without routine rubella immunization, should consider the rubella susceptibility of staff and the risk of hospital-based rubella outbreaks.


General-purpose monitoring during speech production.
The concept of "monitoring" refers to our ability to control our actions on-line. Monitoring involved in speech production is often described in psycholinguistic models as an inherent part of the language system. We probed the specificity of speech monitoring in two psycholinguistic experiments where electroencephalographic activities were recorded. Our focus was on a component previously reported in nonlinguistic manual tasks and interpreted as a marker of monitoring processes. The error negativity (Ne, or error-related negativity), thought to originate in medial frontal areas, peaks shortly after erroneous responses. A component of seemingly comparable properties has been reported, after errors, in tasks requiring access to linguistic knowledge (e.g., speech production), compatible with a generic error-detection process. However, in contrast to its original name, advanced processing methods later revealed that this component is also present after correct responses in visuomotor tasks. Here, we reported the observation of the same negativity after correct responses across output modalities (manual and vocal responses). This indicates that, in language production too, the Ne reflects on-line response monitoring rather than error detection specifically. Furthermore, the temporal properties of the Ne suggest that this monitoring mechanism is engaged before any auditory feedback. The convergence of our findings with those obtained with nonlinguistic tasks suggests that at least part of the monitoring involved in speech production is subtended by a general-purpose mechanism.


PRISMA-S: an extension to the PRISMA Statement for Reporting Literature Searches in Systematic Reviews.
Literature searches underlie the foundations of systematic reviews and related review types. Yet, the literature searching component of systematic reviews and related review types is often poorly reported. Guidance for literature search reporting has been diverse, and, in many cases, does not offer enough detail to authors who need more specific information about reporting search methods and information sources in a clear, reproducible way. This document presents the PRISMA-S (Preferred Reporting Items for Systematic reviews and Meta-Analyses literature search extension) checklist, and explanation and elaboration. The checklist was developed using a 3-stage Delphi survey process, followed by a consensus conference and public review process. The final checklist includes 16 reporting items, each of which is detailed with exemplar reporting and rationale. The intent of PRISMA-S is to complement the PRISMA Statement and its extensions by providing a checklist that could be used by interdisciplinary authors, editors, and peer reviewers to verify that each component of a search is completely reported and therefore reproducible.


Patient-reported outcomes data in patients with psoriatic arthritis from a randomised trial of etanercept and methotrexate as monotherapy or in combination.
We examined patient-reported outcomes (PROs) in The Study of Etanercept And Methotrexate in Patients with Psoriatic Arthritis (PsA); a 48-week, phase 3, randomised controlled trial that compared outcomes with methotrexate (MTX) monotherapy, etanercept monotherapy, and MTX+ etanercept in patients with PsA. Efficacy endpoints included: mean changes from baseline and proportion of patients who reported improvements≥minimal clinically important difference (MCID) at week 24 in treatment groups for Health Assessment Questionnaire-Disability Index, Patient Global Assessment (PtGA), Patient Global Assessment of Joint Pain (PtGAJP) and Medical Outcomes Study Short Form-36 Questionnaire (SF-36) Physical Component Summary (PCS), and Mental Component Summary, and eight domain scores. PROs were analysed as reported (observed), without multiplicity adjustment; therefore, p values are descriptive. At week 24, patients receiving etanercept monotherapy or MTX+ etanercept combination reported greater improvements (p≤0.05) in PtGA, PtGAJP and SF-36 PCS scores compared with those receiving MTX monotherapy. Compared with MTX monotherapy, higher proportions of patients receiving etanercept monotherapy and combination therapy reported improvements≥MCID in PtGA (etanercept vs MTX, p=0.005) and PtGAJP (MTX +etanercept vs MTX, p=0.038). Across PROs, proportions of patients reporting scores≥age and gender-matched normative values at week 24 ranged from 20.8% to 51.0% with MTX monotherapy, 30.9% to 48.8% with etanercept monotherapy, and 30.6% to 52.3% with MTX+ etanercept combination. Patients receiving etanercept monotherapy or MTX+ etanercept reported greater improvements from baseline in several PROs compared with those receiving MTX monotherapy. PROs should be incorporated in discussions between patients and clinicians regarding their treatment choices as they can help determine which treatments are more beneficial in patients with PsA.


Significant effect of polymorphisms in CYP2D6 and ABCC2 on clinical outcomes of adjuvant tamoxifen therapy for breast cancer patients.
The clinical efficacy of tamoxifen is suspected to be influenced by the activity of drug-metabolizing enzymes and transporters involved in the formation, metabolism, and elimination of its active forms. We investigated relationships of polymorphisms in transporter genes and CYP2D6 to clinical outcome of patients receiving tamoxifen. We studied 282 patients with hormone receptor-positive, invasive breast cancer receiving tamoxifen monotherapy, including 67 patients who have been previously reported. We investigated the effects of allelic variants of CYP2D6 and haplotype-tagging single nucleotide polymorphisms (tag-SNPs) of ABCB1, ABCC2, and ABCG2 on recurrence-free survival using the Kaplan-Meier method and Cox regression analysis. Plasma concentrations of tamoxifen metabolites were measured in 98 patients receiving tamoxifen 20 mg/d. CYP2D6 variants were significantly associated with shorter recurrence-free survival (P = .000036; hazard ratio [HR] = 9.52; 95% CI, 2.79 to 32.45 in patients with two variant alleles v patients without variant alleles). Among 51 tag-SNPs in transporter genes, a significant association was found at rs3740065 in ABCC2 (P = .00017; HR = 10.64; 95% CI, 1.44 to 78.88 in patients with AA v GG genotypes). The number of risk alleles of CYP2D6 and ABCC2 showed cumulative effects on recurrence-free survival (P = .000000055). Patients carrying four risk alleles had 45.25-fold higher risk compared with patients with <or= one risk allele. CYP2D6 variants were associated with lower plasma levels of endoxifen and 4-hydroxytamoxifen (P = .0000043 and .00052), whereas no significant difference was found among ABCC2 genotype groups. Our results suggest that polymorphisms in CYP2D6 and ABCC2 are important predictors for the prognosis of patients with breast cancer treated with tamoxifen.


Characterization of CD4 T Cell Epitopes of Infliximab and Rituximab Identified from Healthy Donors.
The chimeric antibodies anti-CD20 rituximab (Rtx) and anti-TNFα infliximab (Ifx) induce antidrug antibodies (ADAs) in many patients with inflammatory diseases. Because of the key role of CD4 T lymphocytes in the initiation of antibody responses, we localized the CD4 T cell epitopes of Rtx and Ifx. With the perspective to anticipate immunogenicity of therapeutic antibodies, identification of the CD4 T cell epitopes was performed using cells collected in healthy donors. Nine T cell epitopes were identified in the variable chains of both antibodies by deriving CD4 T cell lines raised against either Rtx or Ifx. The T cell epitopes often exhibited a good affinity for human leukocyte antigen (HLA)-DR molecules and were part of the peptides identified by MHC-associated peptide proteomics assay from HLA-DR molecules of dendritic cells (DCs) loaded with the antibodies. Two-third of the T cell epitopes identified from the healthy donors stimulated peripheral blood mononuclear cells from patients having developed ADAs against Rtx or Ifx and promoted the secretion of a diversity of cytokines. These data emphasize the predictive value of evaluating the T cell repertoire of healthy donors and the composition of peptides bound to HLA-DR of DCs to anticipate and prevent immunogenicity of therapeutic antibodies.


Carotid stiffness in young adults: a life-course analysis of its early determinants: the Amsterdam Growth and Health Longitudinal Study.
Cardiovascular risk factors affecting arterial stiffness in adulthood may develop at different critical periods earlier in life. We examined whether the trajectories, from adolescence to young adulthood, of blood pressure, body fatness and fat distribution, blood lipids, cardiorespiratory fitness, and heart rate determined levels of arterial stiffness in young adults. We investigated 373 apparently healthy adults in whom cardiovascular risk factors were repeatedly examined between the ages of 13 and 36 years and carotid stiffness estimates were obtained at the age of 36 years. Differences in the mean levels and the trajectories of risk factors throughout the 24-year longitudinal period between subjects with different levels of carotid stiffness at age 36 years were analyzed with generalized estimating equations. Compared with individuals with less stiff carotid arteries, those with stiffer carotid arteries at the age of 36 years were characterized from ages 13 to 36 years by greater levels of and steeper increases in blood pressure and central fatness, independently of each other and other risk factors. These increases were already present in adolescence, preceded the development of poorer levels of blood lipids, cardiorespiratory fitness, and heart rate, which were evident during adulthood only, and explained to a great extent the deleterious association between these risk factors and carotid stiffness at the age of 36 years. Multiple and intertwined mechanisms involved in the pathogenesis of arterial stiffness have their origins in early life. Blood pressure and central fatness have a pivotal role herein and should be specifically targeted to prevent arterial stiffening and its cardiovascular sequelae.


The effect of visual and interaction fidelity on spatial cognition in immersive virtual environments.
Accuracy of memory performance per se is an imperfect reflection of the cognitive activity (awareness states) that underlies performance in memory tasks. The aim of this research is to investigate the effect of varied visual and interaction fidelity of immersive virtual environments on memory awareness states. A between groups experiment was carried out to explore the effect of rendering quality on location-based recognition memory for objects and associated states of awareness. The experimental space, consisting of two interconnected rooms, was rendered either flat-shaded or using radiosity rendering. The computer graphics simulations were displayed on a stereo head-tracked Head Mounted Display. Participants completed a recognition memory task after exposure to the experimental space and reported one of four states of awareness following object recognition. These reflected the level of visual mental imagery involved during retrieval, the familiarity of the recollection, and also included guesses. Experimental results revealed variations in the distribution of participants' awareness states across conditions while memory performance failed to reveal any. Interestingly, results revealed a higher proportion of recollections associated with mental imagery in the flat-shaded condition. These findings comply with similar effects revealed in two earlier studies summarized here, which demonstrated that the less "naturalistic" interaction interface or interface of low interaction fidelity provoked a higher proportion of recognitions based on visual mental images.


The Aperture Capture Illusion: misperceived forms in dynamic occlusion displays.
Visual illusions can reveal unconscious representations and processes at work in perception. Here we report a robust illusion that involves the misperception of moving, partially occluded objects. When a dynamically occluded object is seen through 2 misaligned apertures, the object appears misaligned in the direction of the apertures, creating the Aperture Capture Illusion. Specifically, when part of a dynamically occluded object disappears behind an occluding surface and then another part of the object comes into view immediately afterward, the 2 parts appear misaligned in the direction of the offset of the apertures through which they were seen. This illusion can be nulled: Separating the 2 object parts to increase the time interval between their appearance produced the percept of alignment. The ability to null the illusion in this manner demonstrates that dynamically occluded regions of moving objects continue to persist in perceptual awareness but, we argue, are perceived to move at a slower velocity than visible regions. We report 7 experiments establishing the existence of the illusion and ruling out several classes of explanation for it. We interpret the illusion and the ability to nullify it within the context of Palmer, Kellman, and Shipley's (2006) theory of spatiotemporal object formation.


A Glimpse of Nucleo-Cytoplasmic Large DNA Virus Biodiversity through the Eukaryotic Genomics Window.
The nucleocytoplasmic large DNA viruses (NCLDV) are a group of extremely complex double-stranded DNA viruses, which are major parasites of a variety of eukaryotes. Recent studies showed that certain eukaryotes contain fragments of NCLDV DNA integrated in their genome, when surprisingly many of these organisms were not previously shown to be infected by NCLDVs. We performed an update survey of NCLDV genes hidden in eukaryotic sequences to measure the incidence of this phenomenon in common public sequence databases. A total of 66 eukaryotic genomic or transcriptomic datasets-many of which are from algae and aquatic protists-contained at least one of the five most consistently conserved NCLDV core genes. Phylogenetic study of the eukaryotic NCLDV-like sequences identified putative new members of already recognized viral families, as well as members of as yet unknown viral clades. Genomic evidence suggested that most of these sequences resulted from viral DNA integrations rather than contaminating viruses. Furthermore, the nature of the inserted viral genes helped predicting original functional capacities of the donor viruses. These insights confirm that genomic insertions of NCLDV DNA are common in eukaryotes and can be exploited to delineate the contours of NCLDV biodiversity.


Identification of the copy number variant biomarkers for breast cancer subtypes.
Breast cancer is a common and threatening malignant disease with multiple biological and clinical subtypes. It can be categorized into subtypes of luminal A, luminal B, Her2 positive, and basal-like. Copy number variants (CNVs) have been reported to be a potential and even better biomarker for cancer diagnosis than mRNA biomarkers, because it is considerably more stable and robust than gene expression. Thus, it is meaningful to detect CNVs of different cancers. To identify the CNV biomarker for breast cancer subtypes, we integrated the CNV data of more than 2000 samples from two large breast cancer databases, METABRIC and The Cancer Genome Atlas (TCGA). A Monte Carlo feature selection-based and incremental feature selection-based computational method was proposed and tested to identify the distinctive core CNVs in different breast cancer subtypes. We identified the CNV genes that may contribute to breast cancer tumorigenesis as well as built a set of quantitative distinctive rules for recognition of the breast cancer subtypes. The tenfold cross-validation Matthew's correlation coefficient (MCC) on METABRIC training set and the independent test on TCGA dataset were 0.515 and 0.492, respectively. The CNVs of PGAP3, GRB7, MIR4728, PNMT, STARD3, TCAP and ERBB2 were important for the accurate diagnosis of breast cancer subtypes. The findings reported in this study may further uncover the difference between different breast cancer subtypes and improve the diagnosis accuracy.


Sub-diffraction-limited interference photolithography with metamaterials.
We present that an interference lithography technique beyond the diffraction limit can be theoretically achieved by positing an anisotropic metamaterial under the conventional lithographic mask. Based on the special dispersion characteristics of the metamaterial, only the enhanced evanescent waves with high spatial frequencies can transmit through the metamaterial and contribute to the lithography process. Rigorous coupled wave analysis shows that with 442nm exposure light, one-dimensional periodical structures with 40nm features can be patterned. This technique provides an alternative method to fabricate large-area nanostructures.


Effects of resistance exercise combined with essential amino acid supplementation and energy deficit on markers of skeletal muscle atrophy and regeneration during bed rest and active recovery.
Spaceflight and bed rest (BR) lead to muscle atrophy. This study assessed the effect of essential amino acid (EAA) supplementation and resistance training with decreased energy intake on molecular changes in skeletal muscle after 28-day BR and 14-day recovery. Thirty-one men (31-55 years) subjected to an 8 ± 6% energy deficit were randomized to receive EAA without resistance training (AA, n = 7), or EAA 3 h after (RT, n = 12) or 5 min before (AART, n = 12) resistance training. During BR, myostatin transcript levels increased twofold in the AA group. During recovery, insulin-like growth factor-1 (IGF-1) mRNA increased in all groups, whereas Pax7, MyoD, myogenin, and MRF4 transcripts increased in AA only (all P < 0.05). MAFbx transcripts decreased twofold with AA and RT. Satellite cells did not change during BR or recovery. This suggests that EAA alone is the least protective countermeasure to muscle loss, and several molecular mechanisms are proposed by which exercise attenuates muscle atrophy during BR with energy deficit.


Embryonic stem-cell gametes: the new frontier in human reproduction.
As infertility increases and gamete donations decline, an alternate source of sex cells may prove valuable for research and infertility treatment. This article examines the social and scientific value of gametes derived from the differentiation of established human embryonic stem (ES)-cell lines (ES-cell-derived gametes) and customized gametes created using nuclear transfer technologies to contain a haploid set of genes creating children genetically related to parent(s). ES-cell-derived gametes may be valuable as a resource for biomedical research, instruction and training in assisted reproductive technologies and perhaps for creating children. The creation of children by ES-cell-derived and customized gametes may not result in psychological harm to children but customized gametes may lead to physical harm to children or an accumulation of gene mutations in a population. Although the creation of new types of children using ES-cell gametes provides more reproductive choices to both fertile and infertile individuals, the risk or physical harm to children from customized gametes may be so severe that the scope of reproductive liberty must be limited. Further scientific and ethical analysis of the creation of children by ES-cell gametes is required.


Graphene field effect transistor as a probe of electronic structure and charge transfer at organic molecule-graphene interfaces.
The electronic structure of physisorbed molecules containing aromatic nitrogen heterocycles (triazine and melamine) on graphene is studied using a combination of electronic transport, X-ray photoemission spectroscopy and density functional theory calculations. The interfacial electronic structure and charge transfer of weakly coupled molecules on graphene is found to be governed by work function differences, molecular dipole moments and polarization effects. We demonstrate that molecular depolarization plays a significant role in these charge transfer mechanisms even at submonolayer coverage, particularly for molecules which possess strong dipoles. Electronic transport measurements show a reduction of graphene conductivity and charge carrier mobility upon the adsorption of the physisorbed molecules. This effect is attributed to the formation of additional electron scattering sites in graphene by the molecules and local molecular electric fields. Our results show that adsorbed molecules containing polar functional groups on graphene exhibit different coverage behaviour to nonpolar molecules. These effects open up a range of new opportunities for recognition of different molecules on graphene-based sensor devices.


Use of active management of the third stage of labour in seven developing countries.
To document the use of active management of the third stage of labour for preventing postpartum haemorrhage and to explore factors associated with such use in seven developing countries. Nationally representative samples of facility-based deliveries were selected and observed to determine the use of active management of the third stage of labour and associated factors. Policies on active management were assessed through document review and interviews with relevant professionals. Use of a uterotonic during the third or fourth stages of labour was nearly universal. Correct use of active management of the third stage of labour was found in only 0.5% to 32% of observed deliveries due to multiple deficiencies in practice. In every country except Indonesia, policies regarding active management were conflicting. Developing countries have not targeted decreasing postpartum haemorrhage as an achievable goal; there is little use of active management of the third stage of labour, and policies regarding such management often conflict. Studies are needed to identify the most effective components of active management so that the most efficient package of practices can be promoted.


Urinary Incontinence and Health-Seeking Behavior Among White, Black, and Latina Women.
Fewer than half of women with urinary incontinence (UI) seek care for their condition. Our objective was to qualitatively assess the themes surrounding treatment-seeking behaviors. We conducted 12 focus groups with women and, using purposive sampling, we stratified by racial or ethnic group (white, black, Latina) and by UI frequency. All sessions were transcribed and coded for common themes. Comparative thematic analysis was used to describe similarities and differences among groups. In total, 113 (39 white, 41 black, and 33 Latina) community-dwelling women participated in focus groups. There were no differences in treatment-seeking themes between groups with different UI frequency. However, certain themes emerged when comparing racial/ethnic groups. Women from all groups shared experiences of embarrassment and isolation because of UI, which were impediments to care seeking. White and black women described discussions with close friends or family that led to normalization of symptoms and prevented care seeking. Latina women maintained more secrecy about UI and reported the longest delays in seeking care. Women articulated a higher likelihood of seeking care if they had knowledge of treatment options, but white women were more likely to seek UI-related knowledge compared with black or Latina women. Physician communication barriers were identified in all groups. Despite similar experiences, there are different perceptions about care seeking among white, black, and Latina women. Culturally relevant educational resources that focus on a range of treatment options may improve knowledge and thus improve care-seeking behaviors in women with UI.


Response to intensive therapy steps and to glipizide dose in combination with insulin in type 2 diabetes. VA feasibility study on glycemic control and complications (VA CSDM).
The feasibility study for the VA Cooperative Study on Glycemic Control and Complications in Type 2 Diabetes (VA CSDM) prospectively studied 153 insulin-requiring type 2 diabetes patients, randomized between an intensively treated arm and a standard treatment arm during a mean follow-up of 27 months. The glycemic response to each of the progressive, sequential phases of insulin treatment was assessed, along with the incidence of hypoglycemic reactions and the relative efficacy of different doses of glipizide in combination with fixed doses of insulin. Five medical centers participated; half of the patients were assigned to the intensive treatment arm aiming for normal HbA1c levels. Age of patients was 60 +/- 6 years, duration of diabetes 8 +/- 3 years, and BMI 30.7 +/- 4 kg/m2. A four-step management technique was used, with patients moving to the next step if the operational goals were not met: Phase I, evening intermediate or long-acting insulin; phase II, added day-time glipizide; phase III, two injections of insulin alone; and phase IV, multiple daily insulin injections. Home glucose monitoring measurements were done twice daily and at 3:00 A.M. once a week. Hypoglycemic reactions and home glucose monitoring results were recorded and counted in each of the treatment phases. Baseline HbA1c was 9.3 +/- 1.8%, and fasting plus serum glucose was 11.4 +/- 3.3 mmol/1. Fasting serum glucose fell to near normal in phase I, and remained so in the other treatment phases. An HbA1c separation of 2.1% between the arms was maintained during the course of the study, while the intensive arm kept HbA1c levels below 7.3% (P = 0.001). Most of the decrease in HbA1c occurred with one injection of insulin alone (phase I, -1.4%) or adding day-time glipizide (phase II, -1.9% compared with baseline). HbA1c did not decrease further after substituting two injections of insulin alone, with twice the insulin dose. Multiple daily injections resulted in an additional HbA1c fall (-2.4% compared with baseline). However, two-thirds of the patients were still on one or two injections a day at the end of the study. Changes in home glucose monitoring levels paralleled those of the HbA1c, as did the increments in number of reported hypoglycemic reactions, virtually all either "mild" or "moderate" in character. For the combination of glipizide and insulin (phase II), the only significant effect was obtained with daily doses up to 10 mg a day; there were no significant additional benefits with up to fourfold higher daily doses, and HbA1c levels had an upward trend with doses > 20 mg/day. A simple regime of a single injection of insulin, alone or with glipizide, seemed sufficient to obtain clinically acceptable levels of HbA1c for most obese, insulin-requiring type 2 diabetes patients. Further decrease of HbA1c demanded multiple daily injections at the expense of doubling the insulin dose and the rate of hypoglycemic events. In combination therapy, doses of glipizide > 20 mg/day offered no additional benefit.


Immune Checkpoint Inhibitor Dosing: Can We Go Lower Without Compromising Clinical Efficacy?
In just a few years, immune checkpoint inhibitors have dramatically changed the landscape in oncology, offering durable responses and improved survival for many patients across several tumor types. With more than 3,300 new agents in the immuno-oncology pipeline plus a wide array of combinations being studied, it seems this new era is just getting started. These advances come with a significant caveat: most of the world population does not have access to their benefits, because the yearly cost of a novel anticancer medication can routinely exceed $100,000. There is a large amount of data showing that checkpoint inhibitors have significant activity at doses much lower than those currently approved. We review the evidence for reduced drug dosing as a strategy to increase the number of patients who can be treated and what would be needed to further validate this approach.


Grating-assisted coupling of optical fibers and photonic crystal waveguides.
We propose and analyze a highly efficient method of coupling light from optical fibers to two-dimensional photonic crystal waveguides. Efficient coupling is achieved by positioning of a tapered fiber parallel to the linear defect, where the photonic crystal's cladding functions as a grating coupler and provides field confinement as well. Numerical simulations indicate that better than 90% transmission is possible with a full width at half-magnitude bandwidth of 12nm. It is shown that one can increase the bandwidth by increasing the field overlap between the two waveguides.


Acetaminophen and other risk factors for excessive warfarin anticoagulation.
Warfarin is highly effective in preventing thromboembolism, but increases the risk of hemorrhage, particularly at an international normalized ratio (INR) greater than 4.0. Identifying causes of excessive anticoagulation in clinical practice could help target patients at risk for elevated INRs. To determine causes of INRs greater than 6.0 in a clinical practice setting. Case-control study. Outpatient anticoagulant therapy unit. Outpatients followed up prospectively from April 1995 to March 1996 who had been taking warfarin for more than 1 month, had a target INR of 2.0 to 3.0, and were able to be interviewed within 24 hours of their reported INR. Case patients had INRs greater than 6.0; controls were randomly selected from patients having INRs between 1.7 and 3.3. Factors associated with INRs greater than 6.0, including medication use, recent diet, illness, alcohol consumption, and actual warfarin use. A total of 93 cases and 196 controls were interviewed; they did not differ in age, indication for warfarin, length of therapy, warfarin dose, number of prescription medications, or previous INR or long-term INR variability. Acetaminophen ingestion was independently associated in a dose-dependent manner with having an INR greater than 6.0 (P for trend <.001). For the highest-dose category of acetaminophen intake, 9100 mg/wk or more, the odds of having an INR greater than 6.0 were increased 10-fold (95% confidence interval [CI], 2.6-37.9). Other factors independently associated with an INR greater than 6.0 were new medication known to potentiate warfarin (odds ratio [OR], 8.5; 95% CI, 2.9-24.7), advanced malignancy (OR, 16.4; 95% CI, 2.4-111.0), recent diarrheal illness (OR, 3.5; 95% CI,1.4-8.6), decreased oral intake (OR, 3.6; 95% CI, 1.3-9.7), and taking more warfarin than prescribed (OR, 8.1; 95% CI, 2.2-30.0). Higher vitamin K intake (OR, 0.7; 95% CI, 0.5-0.9) and habitual alcohol consumption of from 1 drink every other day to 2 drinks a day (OR, 0.2; 95% CI, 0.1-0.7) were associated with decreased risk. These data suggest that acetaminophen is an underrecognized cause of overanticoagulation in the outpatient setting. Several other clinically important risk factors were identified. Increased monitoring of INR values when such risk factors are present or modification of the risk factors themselves should reduce the frequency of dangerously high levels of anticoagulation.


Symptomatic benefit of momelotinib in patients with myelofibrosis: Results from the SIMPLIFY phase III studies.
Myelofibrosis (MF)-associated constitutional symptoms can severely impact health-related quality of life. Clinical trials in MF traditionally measure symptom response to treatment as a landmark endpoint of total symptom score (TSS) reduction ≥50% from baseline. However, this dichotomous assessment provides a limited view of clinically relevant symptomatic changes. Herein we evaluated longitudinal change from baseline in TSS over the continuous 24-week period and individual symptom scores to obtain a more comprehensive understanding of symptom benefits experienced by patients with MF receiving therapy. Longitudinal symptom change was evaluated using mixed-effect model repeated measure (MMRM) methodology with individual item-level analyses to complement the interpretation of the landmark symptom results in the completed phase III SIMPLIFY studies of momelotinib in MF. MMRM compared mean change in TSS from baseline with Week 24 using data from all patient visits. Generalized estimating equations were used to estimate item-level odds ratios using multiple predictive imputations for missing data. Momelotinib and ruxolitinib groups reported similar overall symptom improvements, with a TSS difference of <1.5 points between groups for each post-baseline visit in SIMPLIFY-1. In SIMPLIFY-2, the improvement in TSS observed in momelotinib-treated patients was consistent with that observed in SIMPLIFY-1, whereas progressive TSS deterioration was observed with control. Item-level scores were heterogeneous in both studies. A similar and greater proportion of momelotinib-treated patients were categorized as "improved" or "stable" compared with control in SIMPLIFY-1 and SIMPLIFY-2, respectively. Odds ratios for between-group comparison ranged from 0.75 to 1.21 in SIMPLIFY-1, demonstrating similarity in likelihood of symptom improvement. In SIMPLIFY-2, the likelihood of symptom improvement in each item was higher in the momelotinib arm. These findings suggest that momelotinib provides clinically relevant symptom benefits in the JAK inhibitor-naïve and JAK inhibitor-exposed settings.


Race- and sex-specific associations of obesity measures with ischemic stroke incidence in the Atherosclerosis Risk in Communities (ARIC) study.
Studies have suggested differences in the association between obesity and ischemic stroke in black versus white populations. In this study, we explored ischemic stroke risk in relation to a variety of obesity measures by sex and race. Using data from the Atherosclerosis Risk in Communities Study, we obtained information on body mass index, waist circumference, and waist-to-hip ratio from 13 549 black and white participants who were aged 45 to 65 years between 1987 and 1989. All were free of cardiovascular disease and cancer at baseline. Incident strokes over a median follow-up of 16.9 years were ascertained from hospital records. Although crude incidence rates of ischemic stroke varied more than 3-fold by race and sex, the relationship between higher measures of obesity and ischemic stroke risk was positive and linear across all groups. The crude incidence of ischemic stroke was 1.2 per 1000 person-years for white women with the lowest body mass index, ranging up to 8.0 per 1000 person-years for black men with the highest body mass index. Hazard ratios for the highest versus lowest quintile of body mass index, waist circumference, and waist-to-hip ratio ranged from 1.43 to 3.19, indicating increased stroke risk associated with obesity, however it was measured, even after adjustment for potential confounders. Additional adjustment for factors that may mediate the relationship, such as diabetes and hypertension, significantly attenuated the associations, suggesting that these factors may explain much of the stroke risk associated with obesity. Degree of obesity, defined by body mass index, waist circumference, or waist-to-hip ratio, was a significant risk factor for ischemic stroke regardless of sex or race.


Magnetic resonance imaging studies in unipolar depression: systematic review and meta-regression analyses.
Previous meta-analyses of structural MRI studies have shown diffuse cortical and sub-cortical abnormalities in unipolar depression. However, the presence of duplicate publications, recruitment of particular age groups and the selection of specific regions of interest means that there is uncertainty about the balance of current research. Moreover, the lack of systematic exploration of highly significant heterogeneity has prevented the generalisability of finding. A systematic review and random-effects meta-analysis was carried out to estimate effect sizes. Possible publication bias, and the impact of various study design characteristics on the magnitude of the observed effect size were systematically explored. The aim of this study was 1) to include structural MRI studies systematically comparing unipolar depression with bipolar disorder and healthy volunteers; 2) to consider all available structures of interest without specific age limits, avoiding data duplication, and 3) to explore the influence of factors contributing to the measured effect sizes systematically with meta-regression analyses. Unipolar depression was characterised by reduced brain volume in areas involved in emotional processing, including the frontal cortex, orbitofrontal cortex, cingulate cortex, hippocampus and striatum. There was also evidence of pituitary enlargement and an excess of white matter hyperintensity volume in unipolar depression. Factors which influenced the magnitude of the observed effect sizes were differences in methods, clinical variables, pharmacological interventions and sample age.


Dynamic imaging of skeletal muscle contraction in three orthogonal directions.
In this study, a multidimensional strain estimation method using biplane ultrasound is presented to assess local relative deformation (i.e., local strain) in three orthogonal directions in skeletal muscles during induced and voluntary contractions. The method was tested in the musculus biceps brachii of five healthy subjects for three different types of muscle contraction: 1) excitation of the muscle with a single electrical pulse via the musculocutaneous nerve, resulting in a so-called "twitch" contraction; 2) a train of five pulses at 10 Hz and 20 Hz, respectively, to obtain a submaximum tetanic contraction; and 3) voluntary contractions at 30, 60, and 100% of maximum contraction force. Results show that biplane ultrasound strain imaging is feasible. The method yielded adequate performance using the radio frequency data in tracking the tissue motion and enabled the measurement of local deformation in both the vertical direction (orthogonal to the arm) and in the horizontal directions (parallel and perpendicular to direction of the arm) in two orthogonal cross sections of the muscle. The twitch experiments appeared to be reproducible in all three directions, and high strains in vertical (25 to 30%) and horizontal (-20% to -10%) directions were measured. Visual inspection of both the ultrasound data, as well as the strain data, revealed a relaxation that was significantly slower than the force decay. The pulse train experiments nicely illustrated the performance of our technique: 1) similar patterns of force and strain waveforms were found; and 2) each stimulation frequency yielded a different strain pattern, e.g., peak vertical strain was 40% during 10-Hz stimulation and 60% during 20-Hz stimulation. The voluntary contraction patterns were found to be both practically feasible and reproducible, which will enable muscles and more natural contraction patterns to be examined without the need of electrical stimulation.


A comparison of animated versus static images in an instructional multimedia presentation.
Sophisticated three-dimensional animation and video compositing software enables the creation of complex multimedia instructional movies. However, if the design of such presentations does not take account of cognitive load and multimedia theories, then their effectiveness as learning aids will be compromised. We investigated the use of animated images versus still images by creating two versions of a 4-min multimedia presentation on vascular neuroeffector transmission. One version comprised narration and animations, whereas the other animation comprised narration and still images. Fifty-four undergraduate students from level 3 pharmacology and physiology undergraduate degrees participated. Half of the students watched the full animation, and the other half watched the stills only. Students watched the presentation once and then answered a short essay question. Answers were coded and marked blind. The "animation" group scored 3.7 (SE: 0.4; out of 11), whereas the "stills" group scored 3.2 (SE: 0.5). The difference was not statistically significant. Further analysis of bonus marks, awarded for appropriate terminology use, detected a significant difference in one class (pharmacology) who scored 0.6 (SE: 0.2) versus 0.1 (SE: 0.1) for the animation versus stills group, respectively (P = 0.04). However, when combined with the physiology group, the significance disappeared. Feedback from students was extremely positive and identified four main themes of interest. In conclusion, while increasing student satisfaction, we do not find strong evidence in favor of animated images over still images in this particular format. We also discuss the study design and offer suggestions for further investigations of this type.


What sustains long-term adherence to structured physical activity after a cardiac event?
Research addressing methods to sustain long-term adherence to physical activity among older adults is needed. This study investigated the motivations and supports deemed necessary to adhere to a community-based cardiac rehabilitation (CBCR) program by individuals with established coronary heart disease. Twenty-four long-term adherers (15 men, 9 women; age 67.7 ± 16.7 yr) took part in focus-group discussions. Constant comparative analysis supported previous research in terms of the importance of referral procedures, social support, and knowledge of health benefits in influencing uptake and adherence to CBCR. Results also highlighted the routine of a structured class and task-, barrier-, and recovery-specific self-efficacy as necessary to sustain long-term adherence for this specific clinical group. Older adults themselves provide rich information on how to successfully support their long-term adherence to structured exercise sessions. Further research into how to build these components into any exercise program is necessary.


Sociodemographic Study of Danish Individuals Diagnosed with Transsexualism.
Male-to-female (MtF) and female-to-male (FtM) individuals with transsexualism (International Classification of Diseases-10) may differ in core clinical and sociodemographic variables such as age, sexual orientation, marriage and parenthood, school, educational level, and employment. Assessing and understanding the implication of such differences may be a key to developing appropriate and effective treatment and intervention strategies for this group. However, research in the area remains sparse and is often on small populations, making the generalization of results from current studies on individuals diagnosed with transsexualism difficult. (i) To describe and assess key sociodemographic and treatment-related differences between MtF and FtM individuals in a Danish population of individuals diagnosed with transsexualism; (ii) to assess possible implications of such difference, if any, for clinical treatment initiatives for individuals diagnosed with transsexualism. Follow-up of 108 individuals who had permission to undergo sex reassignment surgery (SRS, meaning castration and genital plastic surgery) over a 30-year period from 1978 to 2008 through the Gender Identity Unit in Copenhagen, Denmark. The individuals were identified through Social Security numbers. Clinical and sociodemographic data from medical records were collected. The sex ratio was 1.16:1 (MtF : FtM). Mean age at first referral was 26.9 (standard deviation [SD] 8.8) years for FtM and 30.2 (SD 9.7) for MtF individuals. Compared with MtF, FtM had a significantly lower onset age (before 12 years of age) and lower age when permission for SRS was granted. Further, FtM individuals were significantly more often gynephilic (sexually attracted to females) during research period and less likely to start self-initiated hormonal sex reassignment (SR) (treatment with cross-sex hormones). The MtF and FtM groups did not differ in years of school, educational level, employment, or engagement in marriage and cohabitation. As approximately half of MtF started cross-sex hormonal SR without attending a gender unit, future treatment needs to focus on this group of MtF individuals in order to accommodate the medical risks of self-initiated hormonal treatment.Earlier intervention with adolescents appears necessary since three-quarters of FtM individuals before age 12 had problems with their assigned sex. For both MtF and FtM, we found problems in areas of school, education, and employment and recommend further help in these core areas.


Inhibition of angiogenesis by antioxidant micelles.
Antioxidant micelles capable of scavenging reactive oxygen species (ROS) are prepared from poly(ethylene glycol)-b-poly(dopamine) block copolymers. The micelles inhibit tube formation of human umbilical vein endothelial cells (HUVECs) by scavenging endogenous ROS. Furthermore, the micelles inhibit angiogenesis in the chicken ex ovo chorioallantoic membrane assay. The results show that antioxidant micelles containing catechol moieties may be useful in anti-angiogenic therapy to treat various diseases such as cancer.


[Ecology of Vibrio cholerae serogroup 01 in aquatic environments].
The endemic and seasonal nature of cholera depends upon the survival of Vibrio cholerae 01 in a viable but not necessarily culturable state in ecologic niches in aquatic environments during interepidemic periods. To understand the ecology of V. cholerae it is necessary to know which aquatic ecosystems can harbor it and thus contribute to the endemic presence of cholera in Latin America. This article presents a summary of the ecology of V. cholerae 01, organized according to the abiotic and biotic factors that are relevant to the microbe's survival in aquatic environments. This pathogen finds favorable conditions in waters characterized by moderate salinity, high nutrient content, warm temperature, neutral or slightly alkaline pH, and the presence of aquatic macrophages, phytoplankton, zooplankton, fish, mollusks, and crustaceans. These ecologic conditions are typical of estuaries and coastal swamps, and toxigenic V. cholerae 01 is now considered an autochthonous member of the microbial flora of these environments. The microorganism has also shown the ability to colonize freshwater ecosystems in its viable but not necessarily culturable form, if organic or inorganic substrates that favor its survival are available.


Dynamic deformation of red blood cell in dual-trap optical tweezers.
Three-dimensional dynamic deformation of a red blood cell in a dual-trap optical tweezers is computed with the elastic membrane theory and is compared with the experimental results. When a soft particle is trapped by a laser beam, the particle is deformed depending on the radiation stress distribution whereas the stress distribution on the particle in turn depends on the deformation of its morphological shape. We compute the stress re-distribution on the deformed cell and its subsequent deformations recursively until a final equilibrium state solution is achieved. The experiment is done with the red blood cells in suspension swollen to spherical shape. The cell membrane elasticity coefficient is obtained by fitting the theoretical prediction with the experimental data. This approach allows us to evaluate up to 20% deformation of cell's shape.


The importance of work-up (verification) bias correction in assessing the accuracy of SPECT thallium-201 testing for the diagnosis of coronary artery disease.
Noninvasive testing is often evaluated by the sensitivity and specificity in comparison with a more invasive, but more definitive "gold" standard. However, work-up or verification bias, which occurs when the results of a noninvasive test impact the decision to perform the gold standard invasive test, increases the "observed" sensitivity and decreases the "observed" specificity of the noninvasive test. Most large clinical studies utilizing a noninvasive technique to diagnose coronary artery disease have biases, particularly work-up bias. To obtain more accurate measurements of sensitivity and specificity, we determined the observed sensitivity and specificity of stress (exercise and dipyridamole) single photon emission computed tomographic (SPECT) thallium testing for the detection of coronary artery disease by angiography, and then applied previously published equations to correct for work-up bias. From a computerized data base, reports of 4354 stress SPECT thallium studies from January 1, 1986 through December 31, 1992 were reviewed. All patients with a known history of myocardial infarction or prior coronary angiography were excluded, leaving 2688 patients. From this total, 471 patients underwent coronary angiography within 90 days following stress SPECT thallium testing. Coronary artery disease was defined as a visually assessed stenosis of a coronary artery or a major branch > 50%. Of the 2688 stress SPECT thallium studies, 1265 were normal and 1423 were abnormal. For the 471 patients who underwent catheterization within 90 days following stress SPECT thallium testing. the "observed" sensitivity and specificity were 98 and 14%, respectively. After correction for work-up bias, the corrected sensitivity and specificity were 82 +/- 6% and 59 +/- 2%, respectively. Most studies utilizing noninvasive technologies for the detection of coronary artery disease include patients with known coronary artery disease and have work-up bias as well. By knowing the thallium results of patients with and without catheterization, we were able to correct for work-up bias. These data provide better estimate of the sensitivity and specificity of stress SPECT thallium testing.


Terrestrial ecosystem responses to species gains and losses.
Ecosystems worldwide are losing some species and gaining others, resulting in an interchange of species that is having profound impacts on how these ecosystems function. However, research on the effects of species gains and losses has developed largely independently of one another. Recent conceptual advances regarding effects of species gain have arisen from studies that have unraveled the mechanistic basis of how invading species with novel traits alter biotic interactions and ecosystem processes. In contrast, studies on traits associated with species loss are fewer, and much remains unknown about how traits that predispose species to extinction affect ecological processes. Species gains and losses are both consequences and drivers of global change; thus, explicit integration of research on how both processes simultaneously affect ecosystem functioning is key to determining the response of the Earth system to current and future human activities.


Endocytosis of soluble immune complexes leads to their clearance by FcγRIIIB but induces neutrophil extracellular traps via FcγRIIA in vivo.
Soluble immune complexes (ICs) are abundant in autoimmune diseases, yet neutrophil responses to these soluble humoral factors remain uncharacterized. Moreover, the individual role of the uniquely human FcγRIIA and glycophosphatidylinositol (GPI)-linked FcγRIIIB in IC-mediated inflammation is still debated. Here we exploited mice and cell lines expressing these human neutrophil FcγRs to demonstrate that FcγRIIIB alone, in the absence of its known signaling partners FcγRIIA and the integrin Mac-1, internalizes soluble ICs through a mechanism used by GPI-anchored receptors and fluid-phase endocytosis. FcγRIIA also uses this pathway. As shown by intravital microscopy, FcγRIIA but not FcγRIIIB-mediated neutrophil interactions with extravascular soluble ICs results in the formation of neutrophil extracellular traps (NETs) in tissues. Unexpectedly, in wild-type mice, IC-induced NETosis does not rely on the NADPH oxidase, myeloperoxidase, or neutrophil elastase. In the context of soluble ICs present primarily within vessels, FcγRIIIB-mediated neutrophil recruitment requires Mac-1 and is associated with the removal of intravascular IC deposits. Collectively, our studies assign a new role for FcγRIIIB in the removal of soluble ICs within the vasculature that may serve to maintain homeostasis, whereas FcγRIIA engagement of tissue soluble ICs generates NETs, a proinflammatory process linked to autoimmunity.