Aspartame administered in feed, beginning prenatally through life span, induces cancers of the liver and lung in male Swiss mice.
Aspartame (APM) is a well-known intense artificial sweetener used in more than 6,000 products. Among the major users of aspartame are children and women of childbearing age. In previous lifespan experiments conducted on Sprague-Dawley rats we have shown that APM is a carcinogenic agent in multiple sites and that its effects are increased when exposure starts from prenatal life. The aim of this study is to evaluate the potential of APM to induce carcinogenic effects in mice. Six groups of 62-122 male and female Swiss mice were treated with APM in feed at doses of 32,000, 16,000, 8,000, 2,000, or 0 ppm from prenatal life (12 days of gestation) until death. At death each animal underwent complete necropsy and all tissues and organs of all animals in the experiment were microscopically examined. APM in our experimental conditions induces in males a significant dose-related increased incidence of hepatocellular carcinomas (P < 0.01), and a significant increase at the dose levels of 32,000 ppm (P < 0.01) and 16,000 ppm (P < 0.05). Moreover, the results show a significant dose-related increased incidence of alveolar/bronchiolar carcinomas in males (P < 0.05), and a significant increase at 32,000 ppm (P < 0.05). The results of the present study confirm that APM is a carcinogenic agent in multiple sites in rodents, and that this effect is induced in two species, rats (males and females) and mice (males). No carcinogenic effects were observed in female mice. Am. J. Ind. Med. 53:1197-1206, 2010. © 2010 Wiley-Liss, Inc.


Discrimination: a health hazard for people from refugee and asylum-seeking backgrounds resettled in Australia.
Research has shown that discrimination is harmful to health, but there is relatively little known about discrimination experienced by people from refugee and asylum-seeking backgrounds in resettlement countries and associated health effects. This qualitative-focused mixed methods paper reports on discrimination experienced by refugees and asylum seekers, responses to discrimination, and impacts on health. As part of a broader study of housing, social inclusion and health, surveys were completed by 423 adult refugees and asylum seekers living in South Australia who had been in Australia for up to 7 years. The survey included questions on discrimination based on skin colour, ethnicity and religion, as well as questions on hope, trust, belonging, sense of control and health (including the SF-8). Semi-structured interviews were conducted with 65 survey participants, purposively sampled by visa status, continent and gender, further exploring experiences of discrimination. These and survey open-ended responses were analysed thematically. Twenty-two percent of survey participants reported experiences of discrimination since arriving in Australia (14% in the last year), and 90% of these felt that discrimination had harmed their health. Key settings of discrimination were public transport, within the neighbourhood, and in relation to employment. Those who reported discrimination had significantly worse mental health (p < .000) but not physical health. Discrimination was also associated with less sense of belonging (p = .001), lower levels of trust (p = .038), reduced sense of control (p = .012) and less hope (p = .006). Incidents described in interviews and the open-ended survey responses included incivility, physical assault, and denial of services, experienced across intersecting characteristics of race/ethnicity, religion, gender and visa status. Responses to discrimination spanned affective, cognitive and behavioural dimensions, ranging across types of experience, participant characteristics and context, with most individuals reporting multiple response types. While some of the responses were reported by participants as protective of health, participants' reflections indicated significant negative impacts on mental health in particular. Discrimination featured in the resettlement experiences of a significant number of refugees and asylum seekers, with participants reporting clear negative impacts on mental health. Addressing discrimination is a key resettlement and health issue requiring urgent action.


Identification of calcium-modulating cyclophilin ligand (CAML) as transducer of angiotensin II-mediated nuclear factor of activated T cells (NFAT) activation.
Angiotensin II (Ang II) plays a central role in cardiovascular physiology and disease. Ang II type I receptor (AT1) is thought to mediate most actions of Ang II. A novel AT1 receptor intracellular partner called AT1 receptor-associated protein (ATRAP) was identified, but its exact function has not been elucidated. A yeast two-hybrid screen using ATRAP as bait identified calcium-modulating cyclophilin ligand (CAML) as an ATRAP partner. Yeast two-hybrid and coimmunoprecipitation analysis demonstrated that the N-terminal hydrophilic domain of CAML (amino acids (aa) 1-189) mediates a specific interaction between ATRAP and CAML. Our analysis also showed that aa 40-82 of ATRAP contribute to this interaction. Bioluminescence resonance energy transfer and intracellular colocalization analysis by immunofluorescence in HEK293 cells verified this association within endoplasmic reticulum vesicular structures. Functionally, transcriptional reporter assays and RNA interference ATRAP experiments demonstrated that ATRAP knockdown increased nuclear factor of activated T cells (NFAT) activity. Overexpression of ATRAP decreased Ang II- or CAML-induced NFAT transcriptional activation, whereas an ATRAP-interacting domain of CAML (aa 1-189) sensitized NFAT activation in response to Ang II. These results indicate that CAML is an important signal transducer for the actions of Ang II in regulating the calcineurin-NFAT pathway and suggest that the interaction of CAML with ATRAP may mediate the Ang II actions in vascular physiology.


Hernia recurrence as a problem of biology and collagen.
Usually an abdominal wall hernia is regarded as a mechanical problem with a local defect which has to be closed by technical means. Despite the introduction of several therapeutic improvements, recurrent hernias still appear in 10-15%. Therefore, reasons for a recurrence are discussed in a more fundamental way. It is assumed that a failure mainly depends on the quality of the repair. Correspondingly, in principle, the close causal relationship between the technical component and its failure during time is reflected by an s-shaped outcome curve. In contrast, the configuration of the outcome curve changes markedly if a breakdown is caused by numerous components. Then, the superposition of all incidence curves inevitably leads to a linear decline of the outcome curve without any s-shaped deformation. Regarding outcome curves after hernia repair, the cumulative incidences for recurrences of both incisional and inguinal hernia show a linear rise over years. Considering the configuration of outcome curves of patients with hernia disease, it may therefore be insufficient to explain a recurrence just by a failing technical repair. Rather, biological reasons should be suspected, such as a defective wound healing with impaired scarring process. Recent molecular-biological findings provide increasing evidence of underlying biochemical alterations in patients with recurrent hernia. Until predicting markers to identify patients with an impaired wound healing are available and considering the formation of insufficient scar as the underlying disease, the consequences for every surgical repair should be a supplementary reinforcement with nonabsorbable alloplastic nets as flat meshes with an extensive overlap.


Hypusine modification in eukaryotic initiation factor 5A in rodent cells selected for resistance to growth inhibition by ornithine decarboxylase-inhibiting drugs.
Selection of HTC cells in drugs that inhibit ornithine decarboxylase (ODC) has produced two cell lines, HMOA and DH23A/b, that contain increased amounts of more stable ODC. In addition to alterations in ODC, these cells appear to produce modified eukaryotic initiation factor 5A (eIF-5A) at different rates, a reaction that both requires spermidine and is essential for proliferation. Alterations to the modification of eIF-5A by spermidine cannot be accounted for by changes in eIF-5A protein or modified eIF-5A turnover. Deoxyhypusine synthetase activity is similar in the parental and variant cell lines and is unaltered by growth into plateau phase or by spermidine depletion. The increased rate of eIF-5A modification in DH23A/b cells is due to an increased accumulation of the unmodified eIF-5A precursor. Increased precursor accumulation is not due to increased eIF-5A transcription, but rather it can be attributed to a metabolic accumulation caused by growth under conditions of chronically limiting spermidine. Selection using drugs that inhibit ODC apparently does not cause alterations in the eIF-5A modification pathway. These data support the hypothesis that one of the main effects of spermidine depletion is depletion of the modified eIF-5A pool, and that this is a critical factor in the cytostasis often observed after depletion of cellular polyamines.


Assessment of red blood cell parameters and peripheral smear at different temperatures in case of cold agglutination disease.
Cold agglutination disease (CAD) is characterized by an auto-antibody which is able to agglutinate red blood cells (RBCs) at temperatures lower than that of the body, and subsequently to activate the complement system responsible for lysis of RBCs. Patients show hemolytic anemia of varying degrees of severity, which arise or worsen upon exposure to low temperatures. We describe a case who presented with fever and symptoms of asthenia. His investigations yielded bizarre RBC parameters which led to suspicion of a rare CAD, which was confirmed on reviewing RBC parameters, peripheral smear and direct Coomb's test at different temperatures. Hence, we suggest assessment of bizarre RBC parameters and peripheral smear can help in laboratory testing and diagnosis of CAD. It should also not pose embarrassment in laboratory testing to the pathologist for making an early and accurate diagnosis, thus emphasizing the need for an early treatment of CAD.


Somatostatin inhibits intestinal mucosal mast cell degranulation in normal conditions and during mast cell hyperplasia.
Several studies demonstrate that intestinal mucosal mast cells (IMMC) are modulated by nervous reflexes as well as by intraluminal content. We recently demonstrated that peptones, such as ovalbumin hydrolysate (OVH), induce the release of rat mast cell protease II (RMCP II), indicating IMMC degranulation. The response is due to complex neuroendocrine reflexes. Somatostatin (SS) and its analogues have been used as potential treatments for inflammation in other body systems with contradictory results. The aim of this study was to evaluate if somatostatin could contribute to the reduction of intestinal mucosal mast cell degranulation. Anesthetized rats were prepared for duodenal perfusion and mast cell activation was measured by analysis of RMCP II concentration in the duodenal perfusate. Somatostatin significantly decreased RMCP II concentration in both nonstimulated conditions and after ovalbumin hydrolysate perfusion. However, when somatostatin was given previously to OVH, the peptone still induced a slight increase of RMCP II. Similar effects were observed in animals previously treated with capsaicin. These protocols were repeated in animals infected with Trichinella spiralis, which induces mucosal mast cell hyperplasia. In these cases, somatostatin blocked the effect of OVH, thus, preventing an increase in RMCP II concentration. Fresh frozen tissue sections from the duodenum were processed in an attempt to demonstrate the presence of SS receptors in mast cells using immunofluorescence and Fluo-peptide labeling techniques. Confocal images from duodenum specimens demonstrate the existence of SS receptors in positive cells for RMCP II. Taken together, these results indicate that somatostatin diminishes mast cell activity and in consequence could prevent the intestinal responses to mast cell hyperplasia.


Effects of long-term high-fiber diet on macrovascular changes and lipid and glucose levels in STZ-induced diabetic SD rats.
The effects of long-term high-fiber diet on lipid and glucose levels and the histological changes in the coronary arteries and thoracic aorta in STZ-induced diabetic SD rats were investigated. During the first 4 weeks of the study period, all diabetic rats were given regular chow plus water after which, all were grouped according to the following diet regimen: group II, no added cholesterol and glucomannan; group III, no added cholesterol but with glucomannan supplement, group IV, with added cholesterol but no glucomannan supplement; and group V, with both cholesterol and glucomannan supplements. 15% weight of glucomannan and 1.5% weight of cholesterol in regular rat chow were used as supplements when indicated. Non-diabetic rats which received only regular chow served as the control group (group I). In the 18th week all rats were sacrificed and weight gain, glucose, total cholesterol, HDL-cholesterol, triglyceride and lipid peroxidase concentrations were determined. Selected portions of the heart and thoracic aorta were histologically examined. Weight gain was higher in rats supplemented with glucomannan than in those without glucomannan supplements, but the difference is not significant. A lowering tendency in glucose levels was likewise observed. Furthermore, total cholesterol and HDL-cholesterol levels were lower and higher, respectively in diabetic rats receiving glucomannan. Although the triglyceride levels were similar in all rats, lipid peroxidase levels were significantly lower in rats with high-fiber diet.(ABSTRACT TRUNCATED AT 250 WORDS)


Self-Reported Youth and Adult Exposure to Alcohol Marketing in Traditional and Digital Media: Results of a Pilot Survey.
Alcohol marketing is known to be a significant risk factor for underage drinking. However, little is known about youth and adult exposure to alcohol advertising in digital and social media. This study piloted a comparative assessment of youth and adult recall of exposure to online marketing of alcohol. From September to October 2013, a pilot survey of past 30-day exposure to alcohol advertising and promotional content in traditional and digital media was administered to a national sample of 1,192 youth (ages 13 to 20) and 1,124 adults (ages ≥21) using a prerecruited Internet panel maintained by GfK Custom Research. The weighted proportions of youth and adults who reported this exposure were compared by media type and by advertising and promotional content. Youth were more likely than adults to recall exposure to alcohol advertising on television (69.2% vs. 61.9%), radio (24.8% vs. 16.7%), billboards (54.8% vs. 35.4%), and the Internet (29.7% vs. 16.8%), but less likely to recall seeing advertising in magazines (35.7% vs. 36.4%). Youth were also more likely to recall seeing advertisements and pictures on the Internet of celebrities using alcohol (36.1% vs. 20.8%) or wearing clothing promoting alcohol (27.7% vs. 15.9%), and actively respond (i.e., like, share, or post) to alcohol-related content online. Youth report greater exposure to alcohol advertising and promotional content than adults in most media, including on the Internet. These findings emphasize the need to assure compliance with voluntary industry standards on the placement of alcohol advertising and the importance of developing better tools for monitoring youth exposure to alcohol marketing, particularly on the Internet.


Oral health-related resources - a salutogenic perspective on Swedish 19-year-olds.
The aim was to explore health-oriented resources among 19-year-olds and, specifically, how these resources interact with oral health-related attitudes and behaviour. To represent individuals with various psychosocial environments and socioeconomic areas, the participants were selected from different geographical locations of the Public Dental Service clinics in the county of Jönköping, Sweden. A structured questionnaire was distributed, including the instrument 'sense of coherence', for description of the study group, followed by a semi-structured thematized interview. The qualitative method used for sampling and analyses was grounded theory. Data sampling and analysis were performed in parallel procedures and ended up in a sample of ten informants (five women). In the analysis of interview data, a core category was identified, 'Resources of Wealth and Balance in Life - a Foundation for Healthy Choices', describing the central meaning of the informants' perceptions of resources with an essential beneficial impact on oral health. The core category was built on five themes, which in turn had various subthemes, describing different dimensions of resources interacting with beneficial oral health-related attitudes and behaviour: 'Security-building Resources and Support', 'Driving force and Motivation', 'Maturity and Insight', 'Health Awareness' and 'Environmental influences.' The results elucidate personal and environmental health-oriented resources with influence on oral health-related attitudes and behaviours of young individuals. Such beneficial recourses should be recognized by dental personnel to promote oral health.


Proton magnetic resonance spectroscopy in 22q11 deletion syndrome.
People with velo-cardio-facial syndrome or 22q11 deletion syndrome (22q11DS) have behavioral, cognitive and psychiatric problems. Approximately 30% of affected individuals develop schizophrenia-like psychosis. Glutamate dysfunction is thought to play a crucial role in schizophrenia. However, it is unknown if and how the glutamate system is altered in 22q11DS. People with 22q11DS are vulnerable for haploinsufficiency of PRODH, a gene that codes for an enzyme converting proline into glutamate. Therefore, it can be hypothesized that glutamatergic abnormalities may be present in 22q11DS. We employed proton magnetic resonance spectroscopy ((1)H-MRS) to quantify glutamate and other neurometabolites in the dorsolateral prefrontal cortex (DLPFC) and hippocampus of 22 adults with 22q11DS (22q11DS SCZ+) and without (22q11DS SCZ-) schizophrenia and 23 age-matched healthy controls. Also, plasma proline levels were determined in the 22q11DS group. We found significantly increased concentrations of glutamate and myo-inositol in the hippocampal region of 22q11DS SCZ+ compared to 22q11DS SCZ-. There were no significant differences in levels of plasma proline between 22q11DS SCZ+ and 22q11DS SCZ-. There was no relationship between plasma proline and cerebral glutamate in 22q11DS. This is the first in vivo(1)H-MRS study in 22q11DS. Our results suggest vulnerability of the hippocampus in the psychopathology of 22q11DS SCZ+. Altered hippocampal glutamate and myo-inositol metabolism may partially explain the psychotic symptoms and cognitive impairments seen in this group of patients.


Kidney function, β-cell function and glucose tolerance in older men.
Kidney dysfunction induces insulin resistance, but it is unknown if β cell function is affected. To investigate insulin release (β cell function) and glucose tolerance following a standardized oral glucose tolerance test (OGTT) across kidney function strata. Community-based cohort study from the Uppsala Longitudinal Study of Adult Men (ULSAM). PARTICIPANTS AND MAIN OUTCOME MEASURE: Included were 1015 nondiabetic Swedish men aged 70-71 years. All participants underwent OGTT and euglycaemic hyperinsulinaemic clamp (HEGC) tests, allowing determination of insulin sensitivity, β cell function, and glucose tolerance. Kidney function was estimated by cystatin C-algorithms. Mixed models were used to identify determinants of insulin secretion after the hyperglycemic load. As many as 466 (46%) of participants presented moderate-advanced kidney disease. Insulin sensitivity (by HEGC) decreased across decreasing kidney function quartiles. After the OGTT challenge, however, β cell function indices (area under the curve for insulin release, the estimated first phase insulin release, and the insulinogenic index) were incrementally higher. Neither the oral disposition index nor the 2-h postload glucose tolerance differed across the kidney function strata. Mixed models showed that dynamic insulin release during the OGTT was inversely associated with kidney function, despite the correction for each individual's insulin sensitivity or its risk factors. In older men, β cell function after a hyperglycemic load appropriately compensated the loss in insulin sensitivity that accompanies kidney dysfunction. As a result, the net balance between insulin sensitivity and β cell function was preserved.


Meaningful Effect Sizes, Intraclass Correlations, and Proportions of Variance Explained by Covariates for Planning Two- and Three-Level Cluster Randomized Trials of Social and Behavioral Outcomes.
There is a need for greater guidance regarding design parameters and empirical benchmarks for social and behavioral outcomes to inform assumptions in the design and interpretation of cluster randomized trials (CRTs). We calculated the empirical reference values on critical research design parameters associated with statistical power for children's social and behavioral outcomes, including effect sizes, intraclass correlations (ICCs), and proportions of variance explained by a covariate at different levels (R 2). Children from kindergarten to Grade 5 in the samples from four large CRTs evaluating the effectiveness of two classroom- and two school-level preventive interventions. Teacher ratings of students' social and behavioral outcomes using the Teacher Observation of Classroom Adaptation-Checklist and the Social Competence Scale-Teacher. Two types of effect size benchmarks were calculated: (1) normative expectations for change and (2) policy-relevant demographic performance gaps. The ICCs and R 2 were calculated using two-level hierarchical linear modeling (HLM), where students are nested within schools, and three-level HLM, where students were nested within classrooms, and classrooms were nested within schools. Comprehensive tables of benchmarks and ICC values are provided to inform prevention researchers in interpreting the effect size of interventions and conduct power analyses for designing CRTs of children's social and behavioral outcomes. The discussion also provides a demonstration for how to use the parameter reference values provided in this article to calculate the sample size for two- and three-level CRTs designs.


Exogenous nonphysiologic vs physiologic lipids. Divergent mechanisms for correction of permeability barrier dysfunction.
Although barrier function requires cholesterol, free fatty acids, and ceramides, applications of one or two of these lipids to damaged skin impedes barrier recovery, while equimolar mixtures allow normal recovery. Both incomplete and complete mixtures appear to be internalized within the epidermal nucleated layers, followed by the secretion of abnormal vs normal lamellar body contents, respectively. We compared the ability of complete physiologic lipid mixtures vs a nonmetabolized hydrophobic lipid, petrolatum, to repair the barrier and the requirement for intracellular processing of these lipids within the epidermis. Neat petrolatum, which remains restricted to the stratum corneum, produces more rapid improvement in barrier function than the solvent-dispersed physiologic lipids, and its effects are not altered by coapplication of either monensin or brefeldin A (both from Sigma Chemical Co, St Louis, Mo), known inhibitors of exocytosis and organellogenesis, respectively. In contrast, the physiologic lipids enter the nucleated layers in substantial amounts and require longer to produce barrier recovery. Whereas monensin blocks their ability to facilitate barrier recovery, the physiologic lipids overcome brefeldin A-induced delays in barrier recovery, bypassing the subcellular site of brefeldin A blockade, normalizing both lamellar body contents and intercellular bilayers. While petrolatum remains restricted to the stratum corneum, physiologic lipid mixtures influence barrier recovery after transport to subjacent, nucleated layers, followed by internalization, apparent transport to the distal Golgi apparatus, and incorporation into nascent lamellar bodies.


Is the mental wellbeing of young Australians best represented by a single, multidimensional or bifactor model?
Internationally there is a growing interest in the mental wellbeing of young people. However, it is unclear whether mental wellbeing is best conceptualized as a general wellbeing factor or a multidimensional construct. This paper investigated whether mental wellbeing, measured by the Mental Health Continuum-Short Form (MHC-SF), is best represented by: (1) a single-factor general model; (2) a three-factor multidimensional model or (3) a combination of both (bifactor model). 2220 young Australians aged between 16 and 25 years completed an online survey including the MHC-SF and a range of other wellbeing and mental ill-health measures. Exploratory factor analysis supported a bifactor solution, comprised of a general wellbeing factor, and specific group factors of psychological, social and emotional wellbeing. Confirmatory factor analysis indicated that the bifactor model had a better fit than competing single and three-factor models. The MHC-SF total score was more strongly associated with other wellbeing and mental ill-health measures than the social, emotional or psychological subscale scores. Findings indicate that the mental wellbeing of young people is best conceptualized as an overarching latent construct (general wellbeing) to which emotional, social and psychological domains contribute. The MHC-SF total score is a valid and reliable measure of this general wellbeing factor.


From spiking neurons to rate models: a cascade model as an approximation to spiking neuron models with refractoriness.
A neuron that is stimulated repeatedly by the same time-dependent stimulus exhibits slightly different spike timing at each trial. We compared the exact solution of the time-dependent firing rate for a stochastically spiking neuron model with refractoriness (spike response model) with that of an inhomogeneous Poisson process subject to the same stimulus. To arrive at a mapping between the two models we used alternatively (i) a systematic parameter-free Volterra expansion of the exact solution or (ii) a linear filter combined with nonlinear Poisson rate model (linear-nonlinear Poisson cascade model) with a single free parameter. Both the cascade model and the second-order Volterra model showed excellent agreement with the exact rate dynamics of the spiking neuron model with refractoriness even for strong and rapidly changing input. Cascade rate models are widely used in systems neuroscience. Our method could help to connect experimental rate measurements to the theory of spiking neurons.


Addressing the barriers to driver licensing for Aboriginal people in New South Wales and South Australia.
Low rates of driver licensing have been linked to increased risk of transport-related injury, and reduced access to health services, employment and educational opportunities in the Aboriginal population. This paper reports on how barriers to obtaining a driver licence are being addressed in four Aboriginal communities in New South Wales and South Australia. Qualitative data were collected over a four-month period in 2013. Interviews with Aboriginal and non-Aboriginal stakeholders (n=31) and 11 focus groups with Aboriginal participants (n=46) were analysed thematically using a framework approach. Factors facilitating licensing included: family support, professional lessons, alternative testing and programs that assist with literacy, fines management, financial assistance and access to a supervising driver. Stakeholders recommended raising awareness of existing services and funding community-based service provision to promote access to licensing. Facilitating licence participation requires systemic change and long-term investment to ensure interagency collaboration, service use and sustainability of relevant programs, including job search agencies. Implications for public health: The disadvantage faced by Aboriginal people in driver licensing is a fundamental barrier to participation and a social determinant of health. Understanding the factors that promote licensing is crucial to improving access for under-serviced populations; recommendations provide pragmatic solutions to address licensing disadvantage.


Tissue triage and freezing for models of skeletal muscle disease.
Skeletal muscle is a unique tissue because of its structure and function, which requires specific protocols for tissue collection to obtain optimal results from functional, cellular, molecular, and pathological evaluations. Due to the subtlety of some pathological abnormalities seen in congenital muscle disorders and the potential for fixation to interfere with the recognition of these features, pathological evaluation of frozen muscle is preferable to fixed muscle when evaluating skeletal muscle for congenital muscle disease. Additionally, the potential to produce severe freezing artifacts in muscle requires specific precautions when freezing skeletal muscle for histological examination that are not commonly used when freezing other tissues. This manuscript describes a protocol for rapid freezing of skeletal muscle using isopentane (2-methylbutane) cooled with liquid nitrogen to preserve optimal skeletal muscle morphology. This procedure is also effective for freezing tissue intended for genetic or protein expression studies. Furthermore, we have integrated our freezing protocol into a broader procedure that also describes preferred methods for the short term triage of tissue for (1) single fiber functional studies and (2) myoblast cell culture, with a focus on the minimum effort necessary to collect tissue and transport it to specialized research or reference labs to complete these studies. Overall, this manuscript provides an outline of how fresh tissue can be effectively distributed for a variety of phenotypic studies and thereby provides standard operating procedures (SOPs) for pathological studies related to congenital muscle disease.


Murine B cell leukemia (BCL1): organ distribution and kinetics of growth as determined by fluorescence analysis with an anti-idiotypic antibody.
The life history of a transplantable B cell leukemia (BCL1) that arose spontaneously in a BALB/c mouse is described. Animals bearing this tumor live from 2 to 4 months in apparently good health despite massive splenomegaly and leukemia. Antibody to the idiotype or gamma light chain of the tumor IgM was used in conjunction with the fluorescence-activated cell sorter to identify tumor cells in the BCL1-bearing mice. The results suggest that these cells multiply and differentiate in the spleen and subsequently emigrate to the blood. Tumor cells do not recirculate as evidenced by their failure to enter the thoracic duct or to infiltrate lymph nodes to a significant extent. During tumor growth, a population of T cell blasts appears that may be involved with an immune response against the tumor.


Hemolysis test of a centrifugal pump in a pulsatile mode: the effect of pulse rate and RPM variance.
Centrifugal pumps are generally employed as nonpulsatile blood flow pumps; however, these pumps can produce pulsatile flow by periodically alternating the impeller rotation speed. This study investigates blood trauma due to the effect of pulse frequency and various ranges of pump speed. The hemolysis tests were conducted using the Gyro C1E3 pump. The study was divided into the following categories: Group 1 in a nonpulsatile mode; Group 2 operated at 40 bpm with 30% of speed variance; Group 3, 60 bpm with 30% of speed variance; Group 4, 40 bpm with 70% of speed variance; and Group 5, 60 bpm with 70% of speed variance. A flow rate of 3 L/min and a total pressure head of 200 mm Hg were employed in all groups to simulate a percutaneous cardiopulmonary support condition. There were no significant differences in the hemolysis levels among Groups 1, 2, and 3. However, Groups 4 and 5 exhibited a significantly higher hemolysis rate compared to the other groups. These results indicate that a high rate of speed variance increases hemolysis; however, a range of less than 30% does not affect hemolysis. The pulse rate has no significant effect on hemolysis. In conclusion, the higher speed variance increases the hemolysis level when a pulsatile mode is applied with a centrifugal pump at the given test conditions. However, a speed variance of less than 30% or a pulse rate of less than 60 bpm does not affect hemolysis.


Occupancy patterns in superorganisms: a spin-glass approach to ant exploration.
Emergence of collective, as well as superorganism-like, behaviour in biological populations requires the existence of rules of communication, either direct or indirect, between organisms. Because reaching an understanding of such rules at the individual level can be often difficult, approaches carried out at higher, or effective, levels of description can represent a useful alternative. In the present work, we show how a spin-glass approach characteristic of statistical physics can be used as a tool to characterize the properties of the spatial occupancy patterns of a biological population. We exploit the presence of pairwise interactions in spin-glass models for detecting correlations between occupancies at different sites in the media. Such correlations, we claim, represent a proxy to the existence of planned and/or social strategies in the spatial organization of the population. Our spin-glass approach does not only identify those correlations but produces a statistical replica of the system (at the level of occupancy patterns) that can be subsequently used for testing alternative conditions/hypothesis. Here, this methodology is presented and illustrated for a particular case of study: we analyse occupancy patterns of Aphaenogaster senilis ants during foraging through a simplified environment consisting of a discrete (tree-like) artificial lattice. Our spin-glass approach consistently reproduces the experimental occupancy patterns across time, and besides, an intuitive biological interpretation of the parameters is attainable. Likewise, we prove that pairwise correlations are important for reproducing these dynamics by showing how a null model, where such correlations are neglected, would perform much worse; this provides a solid evidence to the existence of superorganism-like strategies in the colony.


Effects of mobile health interventions on improving glycemic stability and quality of life in patients with type 1 diabetes: A meta-analysis.
This meta-analysis aimed to examine the effects of mobile-health-based (mHealth) interventions on improving glycemic stability and quality of life (QOL) in patients with type 1 diabetes (T1D). Various databases, including PubMed, Embase, CINAHL, Cochrane Library, ProQuest, Chinese Electronic Periodical Services, and China Knowledge Resource Integrated, were used to search for relevant articles. A fixed-effects model or random-effects model was used to examine the overall effect. Various methods, including Egger's test, Begg's test, and trim-and-fill, were adopted to examine publication bias. In total, 26 studies were recruited. Results of the random-effects model showed that the use of mHealth-based interventions significantly decreased glycated hemoglobin (HbA1c) (mean difference = -0.37, 95% confidence interval (CI) = -0.53 to -0.22, p < .001), and improved life satisfaction (Hedges' g = 0.30, 95% CI = 0.10 to 0.50, p = .003), worry of diabetes (Hedges' g = -0.25, 95% CI = -0.41 to 0.08, p = .004), and mental health (Hedges' g = 0.36, 95% CI = 0.08 to 0.64, p = .012). Both adults and youths with T1D can benefit from mHealth-based interventions to improve HbA1c (Hedges' g = -0.44, p = .002 vs. -0.30, p = .003). The effect of mHealth-based interventions on improving QOL in both adults and youths could not be examined due to only one study published in adults with T1D. Moreover, those studies that included the function of feedback from professionals showed a significant effect of decreasing HbA1c compared to those without that function (Hedges' g = -0.48 vs. -0.16, p = .019). Mobile devices are convenient, instantaneous, and easy to use to communicate. Applying mHealth-based interventions with the function of feedback from professionals can be considered an alternative healthcare service to achieve optimal glycemic stability in adults and youths with T1D.


Symptoms of Internet Gaming Disorder in Youth: Predictors and Comorbidity.
Internet gaming disorder (IGD) was included in the Addendum to DSM-5 as a condition for further study. Studies of community samples using a diagnostic interview are lacking, and evaluations of the proposed symptoms, comorbidities, and predictors of IGD are scarce. To provide such information participants in a Norwegian prospective community study were assessed with a clinical interview at age 10 years. Symptoms of other psychiatric disorders were measured with the Child and Adolescent Psychiatric Assessment at ages 8 and 10 (n = 740). Children, parents, and teachers provided information on demographics, temperament, intelligence, executive functions, self-concept, social skills, victimization, emotion regulation, family climate, and parenting. Results indicated that IGD was present in 1.7% (95% confidence interval, 0.7-2.7) of the participants (3.0% boys and 0.5% girls). Factor analysis revealed two factors: heavy involvement and negative consequences. The positive predictive value of withdrawal, tolerance, and unsuccessful attempts to control gaming symptoms to the disorder was low. Symptoms of other common disorders correlated weakly with IGD-symptoms (i.e., from r = 0.07 to r = 0.15). Upon adjusting for gender and gaming at age 8, only limited social and emotion regulation skills at age 8 predicted more age-10 IGD symptoms. In conclusion, IGD is already present in a small percentage of Norwegian 10-year olds. At least three of the proposed symptoms -- withdrawal, tolerance and unsuccessful attempts to control gaming -- merit further study given their weak associations with the disorder. Symptoms of IGD are only marginally associated with symptoms of other psychiatric disorders and only predicted by social skills and emotion regulation deficits.


Transcutaneous bilirubin monitoring of newborns.
Using skin jaundice measurements as an index of infant bilirubin status is subject to a number of uncertainties as described by many investigators. They are illustrated by the experience with a commercial device described here. In view of the widespread need for a quantitative measurement that can replace the visual inspections and subjective evaluations of jaundice being routinely used in clinical settings, such jaundice meters are likely to gain increasing acceptance. We expect future research to include more studies on: (1) reflectance spectra or multiple wavelength measurements for SBI; (2) quality control needed for SBI determinations; (3) the relative significance of physiological factors that influence tissue bilirubin deposits; and (4) the comparative significance of blood versus tissue bilirubin concentrations as true indicators of potential nervous tissue damage.


Gender disparities in acute coronary syndrome: a closing gap in the short-term outcome.
The aim of the present study was to analyze gender disparities in a large cohort of acute coronary syndrome (ACS) patients from the Zurich Acute Coronary Syndrome (Z-ACS) Registry. Gender disparities in ACS were examined. The primary endpoint included in-hospital death rate, and the secondary endpoint major adverse cardiac and cerebrovascular events (MACCEs) at 30-day follow-up. Furthermore, independent predictors for MACCEs and death were identified. In total, 2612 patients with ACS were identified. Out of these, 23% were women. The mean age was higher in women (68.6 ± 12.2; P < 0.001). Troponin-T on admission (1.33 ± 4.64 vs. 1.19 ± 3.04 μg/l; P = 0.002) and N-terminal of the prohormone brain natriuretic peptide on admission (3456.2 ± 7286.7 vs. 1665.6 ± 4800.6 ng/l; P < 0.001) were higher in women compared with men. Single-vessel disease was more common in women (44.9 vs. 39.7%; P = 0.023) and, conversely, multivessel disease was more prevalent in male patients as compared with their female counterparts (59.4 vs. 54.4%; P = 0.029). At discharge, men were more likely prescribed statins (89.4 vs. 85.2%; P = 0.004). Overall mortality and MACCEs were similar for both genders. In women, peak creatine kinase and peak C-reactive protein emerged as independent predictors for MACCEs and SBP on admission, and maximal C-reactive protein and use of glycoprotein IIb/IIIa inhibitors (GPIIb/IIIa) as strong independent predictors for in-hospital death. The present results suggest a closing gap in short-term outcome and improvement in cardiac care between women and men. Nonetheless, differences in treatment strategies continue to exist, particularly pertaining to statin regimens at discharge, which might potentially have a powerful impact on long-term outcomes and gender disparities.


Modifiable clinical and lifestyle factors are associated with elevated alanine aminotransferase levels in newly diagnosed type 2 diabetes patients: results from the nationwide DD2 study.
Current literature lacks data on markers of non-alcoholic fatty liver disease (NAFLD) in newly diagnosed type 2 diabetes mellitus (T2DM) patients. We therefore, conducted a cross-sectional study to examine modifiable clinical and lifestyle factors associated with elevated alanine aminotransferase (ALT) levels as a marker of NAFLD in new T2DM patients. Alanine aminotransferase levels were measured in 1026 incident T2DM patients enrolled in the nationwide Danish Centre for Strategic Research in Type 2 Diabetes (DD2) cohort. We examined prevalence of elevated ALT (>38 IU/L for women and >50 IU/L for men) and calculated prevalence ratios associated with clinical and lifestyle factors using Poisson regression. We examined the association with other biomarkers by linear regression. The median value of ALT was 24 IU/L (interquartile range: 18-32 IU/L) in women and 30 IU/L (interquartile range: 22-41 IU/L) in men. Elevated ALT was found in 16% of incident T2DM patients. The risk of elevated ALT was increased in patients who were <40 years old at diabetes debut [adjusted prevalence ratio (aPR): 1.96, 95% confidence interval (CI): 1.15-3.33], in those with alcohol overuse (>14/>21 drinks per week for women/men) (aPR: 1.60, 95% CI: 1.03-2.50), and in those with no regular physical activity (aPR: 1.42, 95% CI: 1.04-1.93). Obesity and metabolic syndrome per se showed no association with elevated ALT when adjusted for other markers, whereas we found positive associations of ALT with increased C-peptide (β = 0.14, 95% CI: 0.06-0.21) and fasting blood glucose (β = 0.07, 95% CI: 0.03-0.11). Among newly diagnosed T2DM patients, several modifiable clinical and lifestyle factors are independent markers of elevated ALT levels.


Long-term differences in language and cognitive function after childhood exposure to anesthesia.
Over the past decade, the safety of anesthetic agents in children has been questioned after the discovery that immature animals exposed to anesthesia display apoptotic neurodegeneration and long-term cognitive deficiencies. We examined the association between exposure to anesthesia in children under age 3 and outcomes in language, cognitive function, motor skills, and behavior at age 10. We performed an analysis of the Western Australian Pregnancy Cohort (Raine) Study, which includes 2868 children born from 1989 to 1992. Of 2608 children assessed, 321 were exposed to anesthesia before age 3, and 2287 were unexposed. On average, exposed children had lower scores than their unexposed peers in receptive and expressive language (Clinical Evaluation of Language Fundamentals: Receptive [CELF-R] and Expressive [CELF-E]) and cognition (Colored Progressive Matrices [CPM]). After adjustment for demographic characteristics, exposure to anesthesia was associated with increased risk of disability in language (CELF-R: adjusted risk ratio [aRR], 1.87; 95% confidence interval [CI], 1.20-2.93, CELF-E: aRR, 1.72; 95% CI, 1.12-2.64), and cognition (CPM: aRR, 1.69; 95% CI, 1.13-2.53). An increased aRR for disability in language and cognition persisted even with a single exposure to anesthesia (CELF-R aRR, 2.41; 95% CI, 1.40-4.17, and CPM aRR, 1.73; 95% CI, 1.04-2.88). Our results indicate that the association between anesthesia and neuropsychological outcome may be confined to specific domains. Children in our cohort exposed to anesthesia before age 3 had a higher relative risk of language and abstract reasoning deficits at age 10 than unexposed children.


Evaluation of periostin levels in gingival crevicular fluid in association between coronary heart disease and chronic periodontitis.
Periostin is a protein, which is essential for periodontal tissue integrity, development and maturity. The aim of this study was to evaluate the role of gingival crevicular fluid (GCF) periostin levels in the association between coronary heart disease (CHD) and chronic periodontitis (CP). This matched case-control study was conducted on 116 participants. The participants were matched for age, gender, and body mass index and divided into four groups as follows: (1) 29 patients with CHD and sever CP (CHD-CP), (2) 29 patients with CHD and without CP (CHD-H), (3) 29 patients without CHD and with sever CP (H-CP), and (4) 29 healthy participants (H-H). The GCF periostin was collected and evaluated using the enzyme-linked immunosorbent assay (ELASA). Finally, the data were analyzed by analysis of variance using the stata software. Significance was assigned at P < 0.05. The results showed that there was a significant difference in the GCF periostin levels in the four groups (P < 0.05). Moreover, according to the results of the Bonferroni's test, differences in the mean periostin levels were significant (P < 0.001) between CHD-CP and CHD-H, CHD-CP and H-CP, CHD-CP and H-H, CHD-H and H-H, and also between H-CP and H-H. The periostin levels reduced in the CHD patients, especially in the CHD-CP group. The findings reveal a probable role of periostin in the association between CHD and CP.


Type A aortic dissection developing during endovascular repair of an acute type B dissection.
To describe an extension of type B dissection into type A during endoluminal treatment of acute type B dissection. A 57-year-old man with acute type B dissection underwent endovascular repair using 2 Excluder stent-grafts. Before releasing the second stent-graft, the patient complained of mild central chest pain, headache, and visual flashes, but no hemodynamic changes were noticed. An angiogram, performed soon afterwards, showed a retrograde extension of the dissection through the aortic arch involving both the left carotid artery and the ascending aorta. The procedure was completed, and the patient was transported to the operating room where the proximal ascending aorta was replaced. Spiral computed tomographic angiography performed 2 weeks after the procedure showed complete exclusion of the primary entry tear. At 6-month follow-up, the patient was asymptomatic; the left carotid artery dissection had disappeared. Endovascular repair of acute type B dissection is technically feasible, but longer experience will help prevent technical complications.


Double-blind placebo controlled study of alpha-adrenergic receptor antagonists (doxazosin) for treatment of voiding dysfunction in the pediatric population.
We sought to determine whether the alpha-adrenergic antagonist doxazosin could be used as primary therapy in children with voiding dysfunction. Children were assigned to maintain a voiding diary and then randomly divided into a double-blind placebo controlled protocol (0.5 mg doxazosin or placebo). Duplicate uroflow studies with post-void residual evaluations and assessment of dysfunctional voiding scores were performed on initiation and completion of the study. At the conclusion parents were asked to rank the perceived improvement of the urinary incontinence (ie parental subjective perception of improvement). No significant differences between doxazosin (18) and placebo (20) treated patients were found in the number of incontinent days per week, severity of incontinent episodes or alterations in uroflow patterns. Although not significant, 2 findings suggested a beneficial effect of doxazosin over placebo. Specifically, doxazosin decreased the number of incontinent episodes weekly from a median of 18 to 4, while the number of incontinent episodes weekly in the placebo group remained essentially unchanged, decreasing from 15 to 14 (p = 0.13). Doxazosin also improved the dysfunctional voiding scores over placebo, for an improvement of -3 vs 0 points. Further substantiating a doxazosin effect over placebo was the subjective perception of the parents that doxazosin significantly improved urinary continence (p <0.02). Compared to placebo, doxazosin did not demonstrate a significant objective benefit, but produced a significant subjective benefit in the treatment of urinary incontinence secondary to voiding dysfunction.


Methamphetamine-induced changes in the mice hippocampal neuropeptide Y system: implications for memory impairment.
Methamphetamine (METH) is a psychostimulant drug that causes irreversible brain damage leading to several neurological and psychiatric abnormalities, including cognitive deficits. Neuropeptide Y (NPY) is abundant in the mammalian central nervous system (CNS) and has several important functions, being involved in learning and memory processing. It has been demonstrated that METH induces significant alteration in mice striatal NPY, Y(1) and Y(2) receptor mRNA levels. However, the impact of this drug on the hippocampal NPY system and its consequences remain unknown. Thus, in this study, we investigated the effect of METH intoxication on mouse hippocampal NPY levels, NPY receptors function, and memory performance. Results show that METH increased NPY, Y(2) and Y(5) receptor mRNA levels, as well as total NPY binding accounted by opposite up- and down-regulation of Y(2) and Y(1) functional binding, respectively. Moreover, METH-induced impairment in memory performance and AKT/mammalian target of rapamycin pathway were both prevented by the Y(2) receptor antagonist, BIIE0246. These findings demonstrate that METH interferes with the hippocampal NPY system, which seems to be associated with memory failure. Overall, we concluded that Y(2) receptors are involved in memory deficits induced by METH intoxication.


Costs and quality of life by disability among people with multiple sclerosis: a register-based study in Sweden.
Population-based estimates of costs of illness and health-related quality of life, by disability levels among people with multiple sclerosis, are lacking. To estimate the annual costs of illness and health-related quality of life, by disability levels, among multiple sclerosis patients, 21-64 years of age. Microdata from Swedish nationwide registers were linked to estimate the prevalence-based costs of illness in 2013, including direct costs (prescription drug use and specialised healthcare) and indirect costs (calculated using sick leave and disability pension), and health-related quality of life (estimated from the EQ-5D). Disability level was measured by the Expanded Disability Status Scale (EDSS). Among 8906 multiple sclerosis patients, EDSS 0.0-3.5 and 7.0-9.5 were associated with mean indirect costs of SEK 117,609 and 461,357, respectively, whereas direct costs were similar between the categories (SEK 117,423 and 102,714, respectively). Prescription drug costs represented 40% of the costs of illness among multiple sclerosis patients with low EDSS, while among patients with high EDSS more than 80% were indirect costs. Among the 1684 individuals who had reported both EQ-5D and EDSS, the lowest health-related quality of life scores were found among those with a high EDSS. Among people with multiple sclerosis, we confirmed higher costs and lower health-related quality of life in higher disability levels, in particular high indirect costs.


Nuclear envelope localization of LEMD2 is developmentally dynamic and lamin A/C dependent yet insufficient for heterochromatin tethering.
Peripheral heterochromatin in mammalian nuclei is tethered to the nuclear envelope by at least two mechanisms here referred to as the A- and B-tethers. The A-tether includes lamins A/C and additional unknown components presumably INM protein(s) interacting with both lamins A/C and chromatin. The B-tether includes the inner nuclear membrane (INM) protein Lamin B-receptor, which binds B-type lamins and chromatin. Generally, at least one of the tethers is always present in the nuclear envelope of mammalian cells. Deletion of both causes the loss of peripheral heterochromatin and consequently inversion of the entire nuclear architecture, with this occurring naturally in rod photoreceptors of nocturnal mammals. The tethers are differentially utilized during development, regulate gene expression in opposite manners, and play an important role during cell differentiation. Here we aimed to identify the unknown chromatin binding component(s) of the A-tether. We analyzed 10 mouse tissues by immunostaining with antibodies against 7 INM proteins and found that every cell type has specific, although differentially and developmentally regulated, sets of these proteins. In particular, we found that INM protein LEMD2 is concomitantly expressed with A-type lamins in various cell types but is lacking in inverted nuclei of rod cells. Truncation or deletion of Lmna resulted in the downregulation and mislocalization of LEMD2, suggesting that the two proteins interact and pointing at LEMD2 as a potential chromatin binding mediator of the A-tether. Using nuclei of mouse rods as an experimental model lacking peripheral heterochromatin, we expressed a LEMD2 transgene alone or in combination with lamin C in these cells and observed no restoration of peripheral heterochromatin in either case. We conclude that in contrary to the B-tether, the A-tether has a more intricate composition and consists of multiple components that presumably vary, at differing degrees of redundancy, between cell types and differentiation stages.


Three valuable peptides from bee and wasp venoms for therapeutic and biotechnological use: melittin, apamin and mastoparan.
While knowledge of the composition and mode of action of bee and wasp venoms dates back 50 years, the therapeutic value of these toxins remains relatively unexploded. The properties of these venoms are now being studied with the aim to design and develop new therapeutic drugs. Far from evaluating the extensive number of monographs, journals and books related to bee and wasp venoms and the therapeutic effect of these toxins in numerous diseases, the following review focuses on the three most characterized peptides, namely melittin, apamin, and mastoparan. Here, we update information related to these compounds from the perspective of applied science and discuss their potential therapeutic and biotechnological applications in biomedicine.


Functional properties and structural characterization of rice δ(1)-pyrroline-5-carboxylate reductase.
The majority of plant species accumulate high intracellular levels of proline to cope with hyperosmotic stress conditions. Proline synthesis from glutamate is tightly regulated at both the transcriptional and the translational levels, yet little is known about the mechanisms for post-translational regulation of the enzymatic activities involved. The gene coding in rice (Oryza sativa L.) for δ(1)-pyrroline-5-carboxylate (P5C) reductase, the enzyme that catalyzes the second and final step in this pathway, was isolated and expressed in Escherichia coli. The structural and functional properties of the affinity-purified protein were characterized. As for most species, rice P5C reductase was able to use in vitro either NADH or NADPH as the electron donor. However, strikingly different effects of cations and anions were found depending on the pyridine nucleotide used, namely inhibition of NADH-dependent activity and stimulation of NADPH-dependent activity. Moreover, physiological concentrations of proline and NADP(+) were strongly inhibitory for the NADH-dependent reaction, whereas the NADPH-dependent activity was mildly affected. Our results suggest that only NADPH may be used in vivo and that stress-dependent variations in ion homeostasis and NADPH/NADP(+) ratio could modulate enzyme activity, being functional in promoting proline accumulation and potentially also adjusting NADPH consumption during the defense against hyperosmotic stress. The apparent molecular weight of the native protein observed in size exclusion chromatography indicated a high oligomerization state. We also report the first crystal structure of a plant P5C reductase at 3.40-Å resolution, showing a decameric quaternary assembly. Based on the structure, it was possible to identify dynamic structural differences among rice, human, and bacterial enzymes.


Genetic polymorphism and exon changes of the constant regions of the human T-cell rearranging gene gamma.
The genomic nucleotide sequences of the constant-region (C) genes of the human T-cell rearranging gene gamma are given. These sequences show considerable allelic and nonallelic variation. Allelic variants exist at both C gamma 1 and C gamma 2 loci in coding regions (as well as in restriction enzyme sites). Both C gamma genes are in the same transcriptional orientation. Moreover, the organization of the nonallelic C gamma genes reveals some interesting features: the C gamma 1 gene, like the mouse C gamma gene, has three exons, whereas the C gamma 2 gene has four exons, including a duplicated second exon that would create a putative protein with an enlarged constant region. However, these two duplicated exons in C gamma 2 have lost the cysteine residue that is thought to be involved in the interchain disulfide bridge.


Reduction of Coxiella burnetii prevalence by vaccination of goats and sheep, The Netherlands.
Recently, the number of human Q fever cases in the Netherlands increased dramatically. In response to this increase, dairy goats and dairy sheep were vaccinated against Coxiella burnetii. All pregnant dairy goats and dairy sheep in herds positive for Q fever were culled. We identified the effect of vaccination on bacterial shedding by small ruminants. On the day of culling, samples of uterine fluid, vaginal mucus, and milk were obtained from 957 pregnant animals in 13 herds. Prevalence and bacterial load were reduced in vaccinated animals compared with unvaccinated animals. These effects were most pronounced in animals during their first pregnancy. Results indicate that vaccination may reduce bacterial load in the environment and human exposure to C. burnetii.


Identification of efflux pump-mediated multidrug-resistant bacteria by the ethidium bromide-agar cartwheel method.
Bacterial multidrug resistance may be mediated by the overexpression of efflux pumps. Conventional evaluation of efflux activity using efflux pump substrates, such as ethidium bromide, requires specialised instrumentation. The agar-based method, previously reported, has been modified to evaluate as many as twelve bacterial strains and has been termed the ethidium bromide-agar cartwheel method. Agar plates containing different concentrations of ethidium bromide were swabbed with bacterial cultures. The cell efflux capacity increased with increasing ethidium bromide concentration, which produced fluorescence of the bacterial mass. The method was shown to be useful for the detection of efflux activity among multidrug-resistant Gram-negative and Gram-positive clinical isolates, as confirmed by the determination of minimum inhibitory concentration for several antibiotics in the presence of known efflux pump inhibitors. This method may be adapted to the clinical laboratory for the presumptive identification of multidrug-resistant isolates that overexpress efflux pump systems.


Does gender matter in non-hodgkin lymphoma? Differences in epidemiology, clinical behavior, and therapy.
Non-Hodgkin lymphoma (NHL) is one of the most common hematologic malignancies worldwide. The incidence of NHL has been rising for several decades; however, in the last 20 years, it reached a plateau. NHL incidence among males is significantly higher than in females. In addition to gender itself, gravidity has a protective role against NHL occurrence. Gender also matters in terms of NHL clinical characteristics. For example, female predominance was found in three extra-nodal sites (the breast, thyroid, and the respiratory system) occasionally involved in NHL. The diagnosis of NHL during pregnancy is associated with a unique clinical behavior. It is usually diagnosed in the second or third trimester and in advanced stage. Furthermore, the histological subtype is highly aggressive, and reproductive organ involvement is common. The reduced rate of NHL among females may be explained by direct effects of estrogens on lymphoma cell proliferation or by its effect on anti-tumor immune response. Gender has an important role in responsiveness to standard B cell NHL treatment. Among older adults, women benefited more from the addition of the anti-CD20 antibody rituximab to standard chemotherapy regimens. This phenomenon can be explained by the difference in clearance rate of rituximab that was found to be significantly lower among older females than older males. In mantle cell lymphoma, women receiving lenalidomide have higher rates of response. An understanding of the mechanisms responsible for gender-associated NHL differences will ultimately improve the clinical approach, allowing for a more accurate assessment of prognosis and patient-tailored treatment.


Activation of growth hormone short loop negative feedback delays puberty in the female rat.
Implantation of GH in the median eminence of the hypothalamus of 23-day-old female rats tonically inhibited serum GH levels throughout prepubertal development (days 23-36) and depressed GH diurnal pulsatile release. Puberty was significantly delayed in GH-deficient animals, a delay associated with a blunted in vitro ovarian steroidal responsiveness to gonadotropins (particularly estradiol) and a marked decrease in prepubertal uterine weight, as evaluated 4, 7, and 13 days after GH implantation. The prepubertal body weight increase was also depressed. Neither ovarian weight nor serum levels of LH, FSH, PRL, or TSH were consistently altered by the GH implant. In addition, evaluation of pulsatile PRL release in 33-day-old rats revealed no difference between control and GH-deficient animals. Ovarian LH receptor content was lower in GH-implanted rats than in controls, suggesting that a decrease in LH receptors may be one of the mechanisms by which a chronic decrease in serum gH depressed the prepubertal ovarian estradiol and, to a lesser extent, the progesterone response to gonadotropins. A direct ovarian site of action for GH was indicated by the results of experiments in which GH was administered to immature hypophysectomized estrogen-treated rats. The in vitro ovarian progesterone response of the GH-treated animals to both hCG and human FSH was distinctly increased by prior in vivo GH treatment. This effect of GH was not reproduced either by the in vivo administration of LH at a dose calculated by RIA to be contaminating the GH preparation or by FSH at a dose that induced a marked increase in aromatase activity in the same ovaries. GH treatment of hypophysectomized rats failed to affect either aromatase activity or hCG-induced estradiol release, indicating that GH does not directly facilitate the production of estradiol by the ovary. The fact that intact rats with GH implants did not actually show a decrease in the ovarian estradiol response to hCG but, rather, failed to show an increased response at the same time as control animals strongly suggests that ovarian maturation was delayed in GH-deficient rats. It is suggested that, in addition to PRL, GH may also play a role in the process of prepubertal reproductive development by enhancing the steroidal response of the ovary to gonadotropins.


Combination Effects of Hispidin and Gemcitabine via Inhibition of Stemness in Pancreatic Cancer Stem Cells.
Natural products extracted from plants can be potent for developing pharmaceutical products. Hispidin, a polyphenolic compound mainly derived from the medicinal mushroom Phellinus linteus, has been shown to have a therapeutic potential against cancer cells. Pancreatic cancer is one of the most aggressive solid malignancies with high resistance to existing drugs. Cancer stem cells (CSCs) are responsible for chemoresistance. The present study aimed to evaluate the anticancer effects of hispidin on pancreatic CSCs. The cytotoxic effects of hispidin on BxPC-3 and AsPC-1 pancreatic cancer cells and BxPC-3 CD44+ CSCs and the synergistic effects of gemcitabine and hispidin on CSCs were evaluated by a series of in vitro experiments including the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), fluorescence-activated cell sorting, colony forming, Transwell assay, immunocytochemistry, sphere-forming, and western blot assays. Hispidin exerted antitumor effects against both BxPC-3 pancreatic cancer cells and CSCs. Furthermore, it was found that hispidin sensitized pancreatic CSCs to gemcitabine and promoted the therapeutic efficacy of gemcitabine. Hispidin might be a novel chemosensitizer for gemcitabine and a potential synergistic agent for increasing the therapeutic index of gemcitabine as a treatment for pancreatic cancer.


Loss of health related quality of life following low-trauma fractures in the elderly.
To estimate the long-term change in health related quality of life (HRQoL) following low-trauma fractures among individuals receiving home care (HC) services or living in long-term care (LTC) facilities using linked healthcare administrative data from Ontario, Canada. HRQoL was estimated using the Health Utility Index (HUI-2) with the InterRai Minimum Data Set (MDS), a mandatory questionnaire for LTC and HC in the province of Ontario (population 14 million). The HUI-2, a validated HRQoL instrument, allows the calculation of health utility where 0 represents death and 1 the best imaginable health state. For reference, the HUI-2 utility value for Canadians aged 80-84 years is 0.61 and the minimal clinically important difference is 0.03. The MDS was linked to Ontario acute care databases for fiscal years 2007-2011 to identify low-trauma fractures using ICD-10-CA codes. Regression models were used to identify predictors of change in HRQoL from pre-fracture levels to 3 years post fracture for several populations. Low-trauma fractures included hip, humerus, vertebral, wrist, multiple and other. Twenty-three thousand six-hundred fifty-five unique patients with low-trauma fractures were identified with pre- and post-fracture HRQoL assessments, of which 5057 individuals had at least 3 years of follow-up. Compared to patients receiving HC services (N = 3303), individuals residing in LTC (N = 1754) were older, taking more medications, and had more comorbidities. LTC patients had more hip fractures (49 % of total versus 29 %). For all fracture types, HRQoL decreased immediately following fracture. Although levels rebounded after the first month, HRQoL up to 36 months never returned to pre-fracture levels even for non-hip fracture. For both HC and LTC cohorts, clinically important and statistically significant decreases in HUI-2 utility scores were observed 36 months post fracture. Of the 6 HUI-2 domains, mobility had the largest impact on change in HRQoL. Regression analysis indicated that living with a musculoskeletal disorder or a neurological condition and living in LTC were associated with greater decrements in utility following a fracture. Based on the analysis of one of the largest studies on HRQoL to date, among individuals living in LTC facilities or receiving HC services, fractures have a significant permanent impact on HRQoL up to 3 years following fracture.


Characterization and functionality of proliferative human Sertoli cells.
It has long been thought that mammalian Sertoli cells are terminally differentiated and nondividing postpuberty. For most previous in vitro studies immature rodent testes have been the source of Sertoli cells and these have shown little proliferative ability when cultured. We have isolated and characterized Sertoli cells from human cadaveric testes from seven donors ranging from 12 to 36 years of age. The cells proliferated readily in vitro under the optimized conditions used with a doubling time of approximately 4 days. Nuclear 5-ethynyl-2'-deoxyuridine (EdU) incorporation confirmed that dividing cells represented the majority of the population. Classical Sertoli cell ultrastructural features, lipid droplet accumulation, and immunoexpression of GATA-4, Sox9, and the FSH receptor (FSHr) were observed by electron and fluorescence microscopy, respectively. Flow cytometry revealed the expression of GATA-4 and Sox9 by more than 99% of the cells, and abundant expression of a number of markers indicative of multipotent mesenchymal cells. Low detection of endogenous alkaline phosphatase activity after passaging showed that few peritubular myoid cells were present. GATA-4 and SOX9 expression were confirmed by reverse transcription polymerase chain reaction (RT-PCR), along with expression of stem cell factor (SCF), glial cell line-derived neurotrophic factor (GDNF), and bone morphogenic protein 4 (BMP4). Tight junctions were formed by Sertoli cells plated on transwell inserts coated with fibronectin as revealed by increased transepithelial electrical resistance (TER) and polarized secretion of the immunoregulatory protein, galectin-1. These primary Sertoli cell populations could be expanded dramatically in vitro and could be cryopreserved. The results show that functional human Sertoli cells can be propagated in vitro from testicular cells isolated from adult testis. The proliferative human Sertoli cells should have important applications in studying infertility, reproductive toxicology, testicular cancer, and spermatogenesis, and due to their unique biological properties potentially could be useful in cell therapy.


Impact of menstrual and reproductive factors on breast cancer risk in Japan: results of the JACC study.
The incidence of breast cancer among Japanese women, a traditionally low-risk population, has increased substantially. To evaluate the association of reproductive factors with breast cancer risk, we examined 38,159 Japanese women, aged 40-79 years, who responded to a questionnaire on reproductive and other lifestyle factors from 1988 to 1990 in the Japan Collaborative Cohort Study. During an average 7.6 years of follow-up, we documented 151 incidents of breast cancers. Cox proportional hazards modeling was employed to estimate relative risks (RR) and 95% confidence intervals (CI). There was a significant decline in the risk of breast cancer with increasing parity among parous women (trend P=0.01). Women with four or more parities had a 69% lower risk than uniparous women, a reduced risk was also evident among menopausal women. Breast cancer risk tended to rise with increasing age at first delivery (trend P=0.05), the association being very apparent among menopausal women (trend P=0.02). Compared to the women who had their first delivery before age 25, those who delayed this event until after age 34 had an RR of 2.12 (95% CI: 0.72-6.21) and 3.33 (1.07-10.3) among the overall subjects and the menopausal, respectively. There was no apparent association of breast cancer risk with age at menarche or menopause. Our study concerning reproductive risk factors suggests that breast cancer in Japan is similar to that in Western countries, and that reproductive factors, particularly the number of parity and age at first delivery, might be important in the etiology of breast cancer among Japanese women.


Quality of life of children and adolescents from São Paulo: reliability and validity of the Brazilian version of the Pediatric Quality of Life Inventory version 4.0 Generic Core Scales.
To evaluate the reliability and validity of the Brazilian version of the Pediatric Quality of Life Inventory (PedsQL 4.0) Generic Core Scales and measure the quality of life of healthy children and adolescents and patients with rheumatic diseases. We followed the translation methodology proposed by the developer of the original English version of the PedsQL 4.0. The instrument was administered by interviews in two groups: 240 apparently healthy children and adolescents from São Paulo (SP, Brazil) and 105 patients with chronic rheumatic diseases matched by age, as well as their respective parents or caregivers. The parent proxy-report was administered to the children's parents or caregivers separately on the same day. Cronbach's alpha values were between 0.6 and 0.9 for all dimensions, demonstrating adequate internal consistency. Patients with rheumatic diseases reported significantly lower PedsQL scores on all dimensions when compared to the healthy control group (p < 0.0001). Construct validity of the Brazilian Portuguese version of the PedsQL 4.0 was also confirmed. Parent proxy-report of patients with rheumatic diseases highly correlated with child self-report for physical functioning (r = 0.77, p < 0.001) and school functioning (r = 0.73, p < 0.001). Lower correlations were observed for emotional and social functioning (r = 0.40 and 0.59, respectively, p < 0.001). The tool demonstrated reliability, validity, and the administration was fast and easy. Quality of life in patients with rheumatic diseases was significantly lower than in the healthy control group, supporting the necessity of a comprehensive approach to rheumatic disease management, focused on the psychosocial dimensions.


Effect of modulating braille dot height on reading regressions.
It is well known that people who read print or braille sometimes make eye or finger movements against the reading direction. The way these regressions are elicited has been studied in detail by manipulating linguistic aspects of the reading material. Actually, it has been shown that reducing the physical intensity or clarity of the visual input signal can also lead to increased regressions during reading. We asked whether the same might be true in the haptic realm while reading braille. We set the height of braille dots at three different levels (high, medium, and low) and asked adult blind, practiced braille readers to read standardized texts without any repetition of content. The results show that setting the braille dot height near the tactile threshold significantly increased the frequency of regressive finger movements. Additionally, at the lowest braille dot height, braille reading speed significantly diminished. These effects did not occur at braille dot heights that were closer to the height of standard braille (medium and high). We tentatively conclude that this effect may be due to a heightened sense of uncertainty elicited by perception near the threshold that seems to be common to the reading process, independent of the sensory input modality. Furthermore, the described effect may be a feature of a brain area that contributes to the reading process mediated by vision as well as touch.


Increased Fas-mediated apoptosis in polymorphonuclear cells from HIV-infected patients.
Neutrophils represent an important line of innate host defence against invading microorganisms and their functional detriment during HIV infection, including accelerated spontaneous cell death, has been shown to contribute to AIDS development. Neutrophils are susceptible to apoptosis via Fas and an interaction between Fas and FasL was suggested originally as a mechanism to explain constitutive neutrophil apoptosis. We have explored some intracellular pathways leading to PMN apoptosis from 28 HIV-infected patients and 24 healthy volunteers. As previously reported, accelerated spontaneous apoptosis was observed in HIV+ patients, but this did not correlate with viral load. Furthermore, an increase in the level of spontaneous apoptosis was detected in neutrophils from HIV-infected patients following inhibition of ERK, suggesting an impairment of this kinase pathway during the early stages of infection which may contribute to PMN dysfunction. An elevated susceptibility to undergo apoptosis was observed following cross-linking of Fas, which correlated both with viral load and co-expression of Fas/FasL surface molecules. Different mechanisms for spontaneous and Fas-induced apoptosis are proposed which together contribute to the neutropenia and secondary infections observed during the progression to AIDS.


Differential stem- and progenitor-cell trafficking by prostaglandin E2.
To maintain lifelong production of blood cells, haematopoietic stem cells (HSCs) are tightly regulated by inherent programs and extrinsic regulatory signals received from their microenvironmental niche. Long-term repopulating HSCs reside in several, perhaps overlapping, niches that produce regulatory molecules and signals necessary for homeostasis and for increased output after stress or injury. Despite considerable advances in the specific cellular or molecular mechanisms governing HSC-niche interactions, little is known about the regulatory function in the intact mammalian haematopoietic niche. Recently, we and others described a positive regulatory role for prostaglandin E2 (PGE2) on HSC function ex vivo. Here we show that inhibition of endogenous PGE2 by non-steroidal anti-inflammatory drug (NSAID) treatment in mice results in modest HSC egress from the bone marrow. Surprisingly, this was independent of the SDF-1-CXCR4 axis implicated in stem-cell migration. Stem and progenitor cells were found to have differing mechanisms of egress, with HSC transit to the periphery dependent on niche attenuation and reduction in the retentive molecule osteopontin. Haematopoietic grafts mobilized with NSAIDs had superior repopulating ability and long-term engraftment. Treatment of non-human primates and healthy human volunteers confirmed NSAID-mediated egress in other species. PGE2 receptor knockout mice demonstrated that progenitor expansion and stem/progenitor egress resulted from reduced E-prostanoid 4 (EP4) receptor signalling. These results not only uncover unique regulatory roles for EP4 signalling in HSC retention in the niche, but also define a rapidly translatable strategy to enhance transplantation therapeutically.


Theoretical and experimental studies of parameter estimation based on chaos feedback synchronization.
In this paper, we perform theoretical and experimental studies of parameter estimation based on chaos feedback synchronization. The Chua chaotic system is applied as the original system, and its time series of a single variable is applied as a feedback variable to drive a simulative system. The simplex method is employed to minimize the synchronization error in the simulative system to iteratively approach the original system parameters. Furthermore, the effects of feedback function, feedback weight, and noise are investigated. The parameter estimation method based on feedback synchronization shows its robustness to noise perturbation. Finally, the circuit experiment of parameter estimation is implemented in order to examine its practical applicability. The original Chua circuit parameters can be well approached from its recorded time series of a single variable. The results show the parameter estimation method based on feedback synchronization may be capable of estimating real system parameters from a chaotic time series.


A descriptive analysis of overviews of reviews published between 2000 and 2011.
Overviews of systematic reviews compile data from multiple systematic reviews (SRs) and are a new method of evidence synthesis. To describe the methodological approaches in overviews of interventions. Descriptive study. We searched 4 databases from 2000 to July 2011; we handsearched Evidence-based Child Health: A Cochrane Review Journal. We defined an overview as a study that: stated a clear objective; examined an intervention; used explicit methods to identify SRs; collected and synthesized outcome data from the SRs; and intended to include only SRs. We did not restrict inclusion by population characteristics (e.g., adult or children only). Two researchers independently screened studies and applied eligibility criteria. One researcher extracted data with verification by a second. We conducted a descriptive analysis. From 2,245 citations, 75 overviews were included. The number of overviews increased from 1 in 2000 to 14 in 2010. The interventions were pharmacological (n = 20, 26.7%), non-pharmacological (n = 26, 34.7%), or both (n = 29, 38.7%). Inclusion criteria were clearly stated in 65 overviews. Thirty-three (44%) overviews searched at least 2 databases. The majority reported the years and databases searched (n = 46, 61%), and provided key words (n = 58, 77%). Thirty-nine (52%) overviews included Cochrane SRs only. Two reviewers independently screened and completed full text review in 29 overviews (39%). Methods of data extraction were reported in 45 (60%). Information on quality of individual studies was extracted from the original SRs in 27 (36%) overviews. Quality assessment of the SRs was performed in 28 (37%) overviews; at least 9 different tools were used. Quality of the body of evidence was assessed in 13 (17%) overviews. Most overviews provided a narrative or descriptive analysis of the included SRs. One overview conducted indirect analyses and the other conducted mixed treatment comparisons. Publication bias was discussed in 18 (24%) overviews. This study shows considerable variation in the methods used for overviews. There is a need for methodological rigor and consistency in overviews, as well as empirical evidence to support the methods employed.


Bifidobacterium scardovii sp. nov., from human sources.
Five strains of an unusual catalase-negative Gram-positive asporogenous rod-shaped bacterium from human sources were subjected to a polyphasic taxonomic study. The presence of fructose-6-phosphate phosphoketolase, a key enzyme of bifidobacterial hexose metabolism, indicated the strains were members of the genus Bifidobacterium but they did not correspond to any of the recognized species of this genus on the basis of biochemical profiles and whole-cell protein analyses. Comparative 16S rRNA gene sequencing confirmed the placement of the isolates in the genus Bifidobacterium, and demonstrated they represent a hitherto unknown subline within the genus displaying > 5% sequence divergence with recognized species. Based on both phenotypic and phylogenetic criteria, it is proposed that the isolates recovered from human sources be classified as a new species, Bifidobacterium scardovii sp. nov.; the type strain is CCUG 13008T (= DSM 13734T).


The acute care nurse practitioner and management of common health problems: a proposal.
The acute care nurse practitioner, an emerging role in acute healthcare settings, is responsible for managing acutely ill patients with multiple acute and chronic health problems. There are variations in acute care nurse practitioner specialty practices but the complexity and potential progression of the acutely ill patient's status require the use of a multidisciplinary approach to patient care to include knowledge of common health problems. The acute care nurse practitioner curriculum should include course content on common health problems and/or physiological alterations, which should be prerequisites for specialty courses.


Degradation of β-casomorphins and identification of degradation products during yoghurt processing using liquid chromatography coupled with high resolution mass spectrometry.
Liquid chromatography-high resolution mass spectrometry (LC-HRMS) was used to investigate the degradation of β-casomorphin 5 (β-CM5) and β-casomorphin 7 (β-CM7) by Streptococcus thermophilus and/or Lactobacillus delbrueckii ssp. bulgaricus, and to identify the degradation products forming during yoghurt processing. Bovine UHT milk was fermented with: (i) a single strain of L. delbrueckii ssp. bulgaricus, (ii) a single strain of S. thermophilus and (iii) the mixture of S. thermophilus and L. delbrueckii ssp. bulgaricus to pH4.5 and then stored at 4°C for 1 and 7days. Results showed that L. delbrueckii ssp. bulgaricus and/or S. thermophilus completely degraded β-CM5 and β-CM7 upon fermentation to pH4.5 and degradation products were significantly influenced by bacteria strains and storage time. Four peptides, β-CNf60-61 (YP), β-CNf62-63 (FP), β-CNf64-66 (GPI) and β-CNf62-66 (FPGPI) were tentatively identified through high resolution MS/MS experiments; however, it was not possible to confirm if either milk protein or β-casomorphins was a source releasing these peptides. Nonetheless, in this study peptides YP and GPI were released by L. delbrueckii ssp. bulgaricus. This is the first time GPI has been identified and thus future investigation of its bioactivity is warranted.


A widespread family of serine/threonine protein phosphatases shares a common regulatory switch with proteasomal proteases.
PP2C phosphatases control biological processes including stress responses, development, and cell division in all kingdoms of life. Diverse regulatory domains adapt PP2C phosphatases to specific functions, but how these domains control phosphatase activity was unknown. We present structures representing active and inactive states of the PP2C phosphatase SpoIIE from Bacillus subtilis. Based on structural analyses and genetic and biochemical experiments, we identify an α-helical switch that shifts a carbonyl oxygen into the active site to coordinate a metal cofactor. Our analysis indicates that this switch is widely conserved among PP2C family members, serving as a platform to control phosphatase activity in response to diverse inputs. Remarkably, the switch is shared with proteasomal proteases, which we identify as evolutionary and structural relatives of PP2C phosphatases. Although these proteases use an unrelated catalytic mechanism, rotation of equivalent helices controls protease activity by movement of the equivalent carbonyl oxygen into the active site.


Nucleolin interacts with US11 protein of herpes simplex virus 1 and is involved in its trafficking.
Herpes simplex virus type 1 (HSV-1) infection induces profound nucleolar modifications at the functional and organizational levels, including nucleolar invasion by several viral proteins. One of these proteins is US11, which exhibits several different functions and displays both cytoplasmic localization and clear nucleolar localization very similar to that of the major multifunctional nucleolar protein nucleolin. To determine whether US11 interacts with nucleolin, we purified US11 protein partners by coimmunoprecipitations using a tagged protein, Flag-US11. From extracts of cells expressing Flag-US11 protein, we copurified a protein of about 100 kDa that was further identified as nucleolin. In vitro studies have demonstrated that nucleolin interacts with US11 and that the C-terminal domain of US11, which is required for US11 nucleolar accumulation, is sufficient for interaction with nucleolin. This association was confirmed in HSV-1-infected cells. We found an increase in the nucleolar accumulation of US11 in nucleolin-depleted cells, thereby revealing that nucleolin could play a role in US11 nucleocytoplasmic trafficking through one-way directional transport out of the nucleolus. Since nucleolin is required for HSV-1 nuclear egress, the interaction of US11 with nucleolin may participate in the outcome of infection.


Diagnosis and Treatment of Cyclothymia: The "Primacy" of Temperament.
Contrary to DSM-5 definition based on recurrence of low grade hypomanic and depressive symptoms, cyclothymia is better defined in a neurodevelopmental perspective as an exaggeration of cyclothymic temperament. Emotional dysregulation with extreme mood instability and reactivity is the core features of the complex symptomatology. In the present article, we critically reviewed the literature on the diagnosis and treatment of cyclothymia, focusing on the temperamental and neurodevelopmental perspectives. Current epidemiological and clinical research showed the high prevalence and the validity of cyclothymia as a distinct form of bipolarity, frequently associated with multiple comorbidities with anxiety, impulse control, substance use, and so called "personality" disorders. Many patients receive correct diagnosis and treatments after many years of illness, when the superposition of complications reduces the possibility of complete remission. A therapeutic model combining the focus on symptomatic presentations with a temperamental perspective seems to represent an effective approach for cyclothymic patients with complex clinical presentations. Cyclothymic mood instability is an understudied issue despite the evidence of its clinical relevance. Unresolved issues concern its diagnostic delimitation and the possible relationships with emotional dysregulation observed in other neurodevelopmental disorders. We need to confirm the specificity of the disorder and to improve its recognition in early phase of the life, especially in youth. Early recognition means avoiding unnecessary complications and establishing specific treatments and clinical management since the beginning.


Genomic duplication, fractionation and the origin of regulatory novelty.
Having diverged 50 MYA, rice remained diploid while the maize lineage became tetraploid and then fractionated by losing genes from one or the other duplicate region. We sequenced and annotated 13 maize genes (counting the duplicate gene as one gene) on one or the other of the pair of homeologous maize regions; 12 genes were present in one cluster in rice. Excellent maize-rice synteny was evident, but only after the fractionated maize regions were condensed onto a finished rice map. Excluding the gene we used to define homeologs, we found zero retention. Once retained, fractionation (loss of functioning DNA sequence) could occur within cis-acting gene space. We chose a retained duplicate basic leucine zipper transcription factor gene because it was well marked with big, exact phylogenetic footprints (CNSs). Detailed alignments of lg2 and retained duplicate lrs1 to their rice ortholog found that fractionation of conserved noncoding sequences (CNSs) was rare, as expected. Of 30 CNSs, 27 were conserved. The 3 unexpected, missing CNSs and a large insertion support subfunctionalization as a reflection of fractionation of cis-acting gene space and the recent evolution of lg2's novel maize leaf and shoot developmental functions. In general, the principles of fractionation and consolidation work well in making sense of maize gene and genomic sequence data.


A multilocus test of simultaneous divergence across the Isthmus of Panama using snapping shrimp in the genus Alpheus.
The completion of the Panamanian Isthmus is one of the greatest natural experiments in evolution, sending multiple species pairs from a broad range of taxonomic groups on independent evolutionary trajectories. The resulting transisthmian sister species have been used as model systems for examining consequences that accompany cessation of gene flow in formerly panmictic populations. However, variance in pairwise genetic distances of these "geminates" often exceeds expectations, seemingly conflicting with the assumption that separation of populations was contemporaneous with the final closure of the Isthmus. Multilocus datasets and coalescent-based analytical methods can be used to estimate divergence times while accounting for variance in gene divergence that predates isolation, thus removing the need to invoke unequal divergence times. Here we present results from Bayesian analyses of sequence data from seven nuclear and one mitochondrial marker in eight transisthmian species pairs in the snapping shrimp genus Alpheus. Divergence times in two species pairs were shown to occur much earlier than the Isthmus final closure, but much of the variance in pairwise genetic distances from cytochrome oxidase I (COI) was explained when ancestral polymorphisms were accounted for. Results illustrate how coalescent approaches may be more appropriate for dating recent divergences than for estimating ancient speciation events.


OikoBase: a genomics and developmental transcriptomics resource for the urochordate Oikopleura dioica.
We report the development of OikoBase (http://oikoarrays.biology.uiowa.edu/Oiko/), a tiling array-based genome browser resource for Oikopleura dioica, a metazoan belonging to the urochordates, the closest extant group to vertebrates. OikoBase facilitates retrieval and mining of a variety of useful genomics information. First, it includes a genome browser which interrogates 1260 genomic sequence scaffolds and features gene, transcript and CDS annotation tracks. Second, we annotated gene models with gene ontology (GO) terms and InterPro domains which are directly accessible in the browser with links to their entries in the GO (http://www.geneontology.org/) and InterPro (http://www.ebi.ac.uk/interpro/) databases, and we provide transcript and peptide links for sequence downloads. Third, we introduce the transcriptomics of a comprehensive set of developmental stages of O. dioica at high resolution and provide downloadable gene expression data for all developmental stages. Fourth, we incorporate a BLAST tool to identify homologs of genes and proteins. Finally, we include a tutorial that describes how to use OikoBase as well as a link to detailed methods, explaining the data generation and analysis pipeline. OikoBase will provide a valuable resource for research in chordate development, genome evolution and plasticity and the molecular ecology of this important marine planktonic organism.


Cavernous sinus syndrome in dogs and cats: case series (2002-2015).
The cavernous sinus (CS) is a paired venous sinus that runs along either side of the pituitary gland on the floor of the calvarium. Cavernous sinus syndrome (CSS) refers to deficits in more than one of the cranial nerves III, IV, V, and VI, as they are in close association in this region. The purpose of this study was to identify the presenting complaints, neurologic findings, diagnosis, and outcomes in dogs and cats with confirmed cavernous sinus syndrome (CSS). Medical records between 2002 and 2015 were reviewed. Inclusion criteria were neurologic signs consistent with CSS and advanced imaging and/or post-mortem examination. Thirteen dogs and 2 cats were included. Twelve dogs received advanced imaging. Post-mortem examination was performed on 2 cats and 3 dogs. Dogs were 6 -13 years (mean= 10.8 years) of age and comprised of several different breeds. Both cats were male neutered domestic shorthair, ages 3 and 14 years. Presenting complaints included mydriasis (N=4), behavior changes (N=3), hyporexia (N=3), ptosis (N=2), ataxia (N=2), pain (N=2), weakness (N=2), lethargy (N=2), and one each of epiphora, ocular swelling, polydipsia, seizures, facial muscle atrophy, dysphagia, and head tilt. Neurologic signs included ophthalmoparesis/plegia (N=13), reduced/absent pupillary light response (N= 11), mydriasis (N= 10), reduced/absent corneal sensation (N= 7), ptosis (N= 6), reduced facial sensation (N= 2), and enophthalmos (N=1). Thirteen patients had a mass lesion within the cavernous sinus, 6 of which were confirmed neoplastic via histopathology. Median survival time for the 4 patients treated with radiation therapy was 1035 days (range 150-2280). Median survival for the 4 patients that received medical treatment was 360 days (range 7-1260 days), and for the 5 non-treated patients 14 days (range 0-90 days). In conclusion mydriasis and ophthalmoplegia are common signs of CSS. A mass lesion within the CS is the most common cause. Survival time may be improved with radiation therapy.


Effects of selenium deficiency on testicular morphology and function in rats.
For four generations rats were fed a low selenium diet (2-7 micrograms Se kg-1) or the same diet with 250 or 300 micrograms Se kg-1 added as selenite. In male rats of the first generation that had been fed the diets from the age of 20 days onwards, selenium depletion led to slightly delayed testis growth during pubertal development that was compensated for in the later stages of maturation. In adult rats fed the low selenium diet for nearly a year no changes in testicular mass and morphology were observed. The serum concentration of testosterone of 6-month-old, selenium-depleted animals was, however, slightly lower than that of adequately supplied controls, and the stimulation of testosterone secretion by administration of GnRH or LH resulted in a significantly less marked rise in the serum concentration of testosterone. From the second generation onwards the testis mass, expressed as a percentage of the body mass, decreased and in the fourth generation was less than 50% of that of the controls. The male gonads of fourth generation animals showed a severe bilateral atrophy, in which the seminiferous tubules were considerably reduced in diameter and almost entirely lined by Sertoli cells and a few stem cells. Differentiated spermatozoa could not be detected. The alterations were reversible and spermatogenesis was restored by feeding the selenium-adequate diet. The findings indicate that testicular morphology and functions are affected by severe selenium deficiency and that the element is necessary for testosterone biosynthesis and the formation and normal development of spermatozoa.