The role of CD80/CD86 in generation and maintenance of functional virus-specific CD8+ T cells in mice infected with lymphocytic choriomeningitis virus.
Lymphocytic choriomeningitis virus (LCMV)-specific CD8(+) T cell responses are considered to be independent of CD28-B7 costimulation. However, the LCMV-specific response has never been evaluated in B7.1/B7.2(-/-) mice. For this reason, we decided to study the T cell response in B7.1/B7.2(-/-) mice infected with two different strains of LCMV, one (Traub strain) typically causing low-grade chronic infection, and another (Armstrong clone 53b) displaying very limited capacity for establishing chronic infection. Using Traub virus we found that most B7.1/B7.2(-/-) mice were unable to rid themselves of the infection. Chronic infection was associated with a perturbed CD8(+) T cell epitope hierarchy, as well as with the accumulation of cells expressing markers of terminal differentiation and being unable to respond optimally to Ag restimulation. Examination of matched CD28(-/-) mice revealed a similar albeit less pronounced pattern of CD8(+) T cell dysfunction despite lack of virus persistence. Finally, analysis of B7.1/B7.2(-/-) mice infected with Armstrong virus revealed a scenario quite similar to that in Traub infected CD28(-/-) mice; that is, the mice displayed evidence of T cell dysfunction, but no chronic infection. Taken together, these results indicate that B7 costimulation is required for induction and maintenance of LCMV-specific CD8(+) T cell memory, irrespective of the LCMV strain used for priming. However, the erosion of CD8(+) T cell memory in B7.1/B7.2(-/-) mice was more pronounced in association with chronic infection. Finally, virus-specific T cell memory was more impaired in the absence of B7 molecules than in the absence of the CD28 receptor, supporting earlier data suggesting the existence of additional stimulatory receptors for B7.


Power and empowerment in nursing: a fourth theoretical approach.
This paper is a discussion of the use of poststructuralism as a means of exploring power and empowerment in nursing. Power and empowerment are well-researched areas of nursing practice, but the issue of how to empower nurses and patients continues to cause debate. Power and empowerment are complex issues and other researchers have provided some clarity by proposing three theoretical approaches: critical social theory, organizational theory and social psychological theory. We support their work and propose an additional poststructural approach as a means of analyzing power and empowerment in nursing. The concept of power in nursing may be critiqued by drawing on the work of Michel Foucault and paying particular attention to two areas: disciplinary power and knowledge/power relationships. Foucault's contention was that behaviour is standardized through disciplinary power and that power and knowledge are intertwined. Nurses who seek an understanding of empowerment must first grasp such workings as hierarchical observation, normalizing judgement, the examination, and power/knowledge relationships, and that cognizance of such issues can promote nursing practice that is empowering. They need to adopt a more critical stance to understanding power and empowerment in nursing, and one way of fostering such criticism is to view nursing practice through a poststructural lens. A poststructural approach merits a place alongside other approaches to understanding power and empowerment in nursing.


Induction of tolerance against the arthritogenic antigen with type-II collagen peptide-linked soluble MHC class II molecules.
In murine collagen-induced arthritis (CIA), self-reactive T cells can recognize peptide antigens derived from type-II collagen (CII). Activation of T cells is an important mediator of autoimmune diseases. Thus, T cells have become a focal point of study to treat autoimmune diseases. In this study, we evaluated the efficacy of recombinant MHC class II molecules in the regulation of antigen-specific T cells by using a self peptide derived from CII (CII260-274; IAGFKGEQGPKGEPG) linked to mouse I-A(q) in a murine CIA model. We found that recombinant I-A(q)/CII260-274 molecules could be recognized by CII-specific T cells and inhibit the same T cells in vitro. Furthermore, the development of CIA in mice was successfully prevented by in vivo injection of recombinant I-A(q)/CII260-274 molecules. Thus, treatment with recombinant soluble MHC class II molecules in complex with an immunodominant self-peptide might offer a potential therapeutic for chronic inflammation in autoimmune disease such as rheumatoid arthritis. [BMB Reports 2016; 49(6): 331-336].


Exercise training and heart rate variability in older people.
Heart rate variability (HRV), a characteristic that is potentially increased by physical activity, has been associated with incidence of cardiac events and total mortality. Since the incidence of cardiac events among older people is high and their physical activity levels and HRV are generally low, it is important to investigate whether regular physical activity can modify HRV in this age group. The purpose of the study was to investigate the effect of regular physical activity on HRV in older men and women. In a randomized controlled trial, the effect of six months' training on HRV was investigated in a group of 51 older men and women (67.0 +/- 5.1 yr). The training group gathered three times per week for 45 min supervised training. At the end of the intervention period, HRV was higher primarily during the day. During daytime, the SD of all normal intervals (+6%) as well as the low frequency component (+ 15%) and the very low frequency component (+ 10%) of HRV were significantly increased (P < 0.05) as compared with the control group. Effects of training were most pronounced in subjects inactive in sports at baseline. This study demonstrates that regular physical activity increases HRV (specifically in the very low and low frequency components) in older subjects. Hence, in older subjects, physical training may be an effective means to modify positively a factor that is associated with increased incidence of cardiac events.


Bridging knowledge to develop an action plan for integrated care for chronic diseases in Greece.
The health, social and economic impact of chronic diseases is well documented in Europe. However, chronic diseases threaten relatively more the 'memorandum and peripheral' Eurozone countries (i.e., Greece, Spain, Portugal and Ireland), which were under heavy recession after the economic crisis in 2009. Especially in Greece, where the crisis was the most severe across Europe, the austerity measures affected mainly people with chronic diseases. As a result, the urgency to tackle the threat of chronic diseases in Greece by promoting public health and providing effective chronic care while flattening the rising health care expenditure is eminent. In many European countries, integrated care is seen as a means to achieve this. The aim of this paper was to support Greek health policy makers to develop an action plan from 2015 onwards, to integrate care by bridging local policy context and needs with knowledge and experience from other European countries. To achieve this aim, we adopted a conceptual framework developed by the World Health Organization on one hand to analyse the status of integrated care in Greece, and on the other to develop an action plan for reform. The action plan was based on an analysis of the Greek health care system regarding prerequisite conditions to integrate care, a clear understanding of its context and successful examples of integrated care from other European countries. This study showed that chronic diseases are poorly addressed in Greece and integrated care is in embryonic stage. Greek policy makers have to realise that this is the opportunity to make substantial reforms in chronic care. Failing to reform towards integrated care would lead to the significant risk of collapse of the Greek health care system with all associated negative consequences. The action plan provided in this paper could support policy makers to make the first serious step to face this challenge. The details and specifications of the action plan can only be decided by Greek policy makers in close cooperation with other health and social care partners. This is the appropriate time for doing so.


Event-Triggered Output-Feedback Adaptive Control of Interconnected Nonlinear Systems: A Cyclic-Small-Gain Approach.
This article proposes a novel output-feedback event-triggered control protocol for a class of interconnected parametric nonlinear systems. Different from most existing works where event-triggering mechanisms are considered for only the controller-to-actuator channel or the sensor-to-controller channel, this work adopts event-triggering mechanisms for both channels as well as adaptive laws. An adaptive states observer is designed to estimate the unavailable system state and then a novel adaptive event-triggered controller is proposed based on the celebrated backstepping technique. By utilizing the cyclic small-gain theorem and Lyapunov theory, it is shown that the proposed control scheme ensures that all the closed-loop signals are semi-globally uniformly ultimately bounded (SGUUB), and the Zeno behavior is successfully excluded. Finally, a practical example is provided to illustrate the effectiveness and advantages of the proposed approach.


Genetic diversity of Echinococcus multilocularis in red foxes in Poland: the first report of a haplotype of probable Asian origin.
The aim of the present study was to estimate the genetic diversity of the cestode Echinococcus multilocularis Leuckart, 1863 in Poland based on sequence analysis of the mitochondrial genes of worms isolated from red foxes, Vulpes vulpes (Linnaeus). Overall, 83 adults of E. multilocularis from the same number of foxes in different parts of Poland were used for analysis. Sequences of the three mitochondrial genes, cytochrome b (cob), NADH dehydrogenase subunit 2 (nad2) and cytochrome c oxidase subunit 1 (cox1), were analysed. Seventy-four individual biological samples were successfully sequenced. Combined sequence analysis of these three genes exhibited fifteen Polish haplotypes (EmPL1-EmPL15). Most isolates (n = 29; 39%) were classified to the EmPL1 haplotype, which occurred mainly in the east, north and centre of Poland. Haplotype EmPL4 (n = 14; 19%) and other haplotypes appeared predominantly in the south and west area. Fourteen haplotypes were grouped in the European clade. One Polish haplotype (EmPL9) (n = 7, 10%) was assigned to the Asian clade with haplotypes from Japan and Kazakhstan. This haplotype was found only in northeast Poland and this is the westernmost report of haplotype of E. multilocularis belonging to the Asian clade in Europe. The investigation demonstrated that populations of E. multilocularis in Poland (and probably also in eastern Europe) included not only different European haplotypes but also those of the Asian origin.


Structural patterns of selection and diversity for Plasmodium vivax antigens DBP and AMA1.
Plasmodium vivax is a significant contributor to the global malaria burden, and a vaccine targeting vivax malaria is urgently needed. An understanding of the targets of functional immune responses during the course of natural infection will aid in the development of a vaccine. Antibodies play a key role in this process, with responses against particular epitopes leading to immune selection pressure on these epitopes. A number of techniques exist to estimate levels of immune selection pressure on particular epitopes, with a sliding window analysis often used to determine particular regions likely to be under immune pressure. However, such analysis neglects protein three-dimensional structural information. With this in mind, a newly developed tool, BioStructMap, was applied to two key antigens from Plasmodium vivax: PvAMA1 and PvDBP Region II. This tool incorporates structural information into tests of selection pressure. Sequences from a number of populations were analysed, examining spatially-derived nucleotide diversity and Tajima's D over protein structures for PvAMA1 and PvDBP. Structural patterns of nucleotide diversity were similar across all populations examined, with Domain I of PvAMA1 having the highest nucleotide diversity and displaying significant signatures of immune selection pressure (Tajima's D > 0). Nucleotide diversity for PvDBP was highest bordering the dimerization and DARC-binding interface, although there was less evidence of immune selection pressure on PvDBP compared with PvAMA1. This study supports previous work that has identified Domain I as the main target of immune-mediated selection pressure for PvAMA1, and also supports studies that have identified functional epitopes within PvDBP Region II. The BioStructMap tool was applied to leading vaccine candidates from P. vivax, to examine structural patterns of selection and diversity across a number of geographic populations. There were striking similarities in structural patterns of diversity across multiple populations. Furthermore, whilst regions of high diversity tended to surround conserved binding interfaces, a number of protein regions with very low diversity were also identified, and these may be useful targets for further vaccine development, given previous evidence of functional antibody responses against these regions.


Quantification of changes in language-related brain areas in autism spectrum disorders using large-scale network analysis.
Diagnosis of autism spectrum disorders (ASD) is difficult, as symptoms vary greatly and are difficult to quantify objectively. Recent work has focused on the assessment of non-invasive diffusion tensor imaging-based biomarkers that reflect the microstructural characteristics of neuronal pathways in the brain. While tractography-based approaches typically analyze specific structures of interest, a graph-based large-scale network analysis of the connectome can yield comprehensive measures of larger-scale architectural patterns in the brain. Commonly applied global network indices, however, do not provide any specificity with respect to functional areas or anatomical structures. Aim of this work was to assess the concept of network centrality as a tool to perform locally specific analysis without disregarding the global network architecture and compare it to other popular network indices. We create connectome networks from fiber tractographies and parcellations of the human brain and compute global network indices as well as local indices for Wernicke's Area, Broca's Area and the Motor Cortex. Our approach was evaluated on 18 children suffering from ASD and 18 typically developed controls using magnetic resonance imaging-based cortical parcellations in combination with diffusion tensor imaging tractography. We show that the network centrality of Wernicke's area is significantly (p<0.001) reduced in ASD, while the motor cortex, which was used as a control region, did not show significant alterations. This could reflect the reduced capacity for comprehension of language in ASD. The betweenness centrality could potentially be an important metric in the development of future diagnostic tools in the clinical context of ASD diagnosis. Our results further demonstrate the applicability of large-scale network analysis tools in the domain of region-specific analysis with a potential application in many different psychological disorders.


Structural brain magnetic resonance imaging of limbic and thalamic volumes in pediatric bipolar disorder.
Youths with bipolar disorder are ideal for studying illness pathophysiology given their early presentation, lack of extended treatment, and high genetic loading. Adult bipolar disorder MRI studies have focused increasingly on limbic structures and the thalamus because of their role in mood and cognition. On the basis of adult studies, the authors hypothesized a priori that youths with bipolar disorder would have amygdalar, hippocampal, and thalamic volume abnormalities. Forty-three youths 6-16 years of age with DSM-IV bipolar disorder (23 male, 20 female) and 20 healthy comparison subjects (12 male, eight female) similar in age and sex underwent structured and clinical interviews, neurological examination, and cognitive testing. Differences in limbic and thalamic brain volumes, on the logarithmic scale, were tested using a two-way (diagnosis and sex) univariate analysis of variance, with total cerebral volume and age controlled. The subjects with bipolar disorder had smaller hippocampal volumes. Further analysis revealed that this effect was driven predominantly by the female bipolar disorder subjects. In addition, both male and female youths with bipolar disorder had significantly smaller cerebral volumes. No significant hemispheric effects were seen. These findings support the hypothesis that the limbic system, in particular the hippocampus, may be involved in the pathophysiology of pediatric bipolar disorder. While this report may represent the largest MRI study of pediatric bipolar disorder to date, more work is needed to confirm these findings and to determine if they are unique to pediatric bipolar disorder.


Cyanocobalamin [c-lactam] inhibits vitamin B12 and causes cytotoxicity in HL60 cells: methionine protects cells completely.
The [c-lactam] derivative of cobalamin antagonizes vitamin B12 in vivo. Therefore, we investigated its effects in tissue culture to develop a model in which to study vitamin B12-deficient hemopoiesis. HL60 cells were cultured in medium containing either methionine or L-homocysteine thiolactone, and various concentrations of 5-methyltetrahydrofolate or pteroylglutamic acid. In medium with L-homocysteine thiolactone, 5-methyltetrahydrofolate, and dialyzed serum, cyanocobalamin [c-lactam] caused cell death, reversible by additional vitamin B12. Pteroylglutamic acid did not prevent this cytotoxic effect. Methionine completely protected cells against cyanocobalamin [c-lactam] for periods of up to 4 months of culture, irrespective of the folate source. Cyanocobalamin [c-lactam] reversibly impaired the incorporation of 5-[14CH3]-tetrahydrofolate and [1-(14)C] propionic acid by intact cells, consistent with inhibition of methionine synthase and methylmalonyl-CoA mutase. A substantial proportion of 5-[14CH3]-tetrahydrofolate uptake could not be suppressed by methionine and may, therefore, have occurred outside of the methionine synthase pathway. These findings are the first indication that cyanocobalamin [c-lactam] antagonizes vitamin B12 in vitro and causes cell death from methionine deficiency. The model should be valuable for investigating the biochemical pathology of vitamin B12-deficient hemopoiesis. The results suggest that methylfolate is not trapped when methionine synthase is inhibited in HL60 cells, but they do not disprove the methylfolate trap hypothesis as applied to normal blood cells.


Depressed interleukin-12 production by peripheral blood mononuclear cells after in vitro stimulation with the 30-kDa antigen in recurrent pulmonary tuberculosis patients.
Some patients develop recurrent tuberculosis (R-TB), even after successfully completing initial anti-tubercular treatment. Although R-TB may be caused by relapse or exogenous reinfection, little is known about the underlying host responses associated with R-TB. This study investigated the profile of cytokines [interferon (IFN)-gamma, interleukin (IL)-12, tumor necrosis factor (TNF)-alpha, IL-6, and IL-10] present in peripheral blood mononuclear cells (PBMCs) of 17 R-TB patients after stimulation with the 30-kDa antigen (Ag) or purified protein derivative (PPD) Ag of Mycobacterium tuberculosis. These data were compared with data obtained from 15 patients with newly diagnosed pulmonary TB (N-TB), 22 patients with treatment failure (TF-TB), and 19 healthy tuberculin reactors (HTR). N-TB and R-TB patients were enrolled in this study within 1 month of beginning anti-tubercular chemotherapy. ELISA results showed that IFN-gamma production following stimulation with the 30-kDa Ag was significantly lower in each group of TB patients than in the HTR controls. In addition, patients with R-TB showed the most significant IL-12 depression among the subject groups after in vitro stimulation with either Ag. Furthermore, a significant decrease in TNF-alpha and IL-10 levels was observed in R-TB patients relative to N-TB patients. However, there was no statistical difference in TNF-alpha and IL-10 production between R-TB patients, TF-TB patients, and HTR controls. Our findings suggest that the underlying mechanisms of cytokine regulation might differ between N-TB and R-TB patients, and that decreased IL-12 production in response to the 30-kDa or PPD Ag might be involved in the immunopathogenesis of human R-TB.


How long is long-term? Reflections based on over 20 years of post-Chernobyl management in Norway.
The radiation protection community has only recently started the important work of preparedness for long-term post-accidental management of radioactively contaminated areas, like for instance the EC projects STRATEGY, FARMING and EURANOS and the French authorities' CODIRPA and PAREX programmes. There are, however, different views concerning how long a long-term management might last. Based on the Norwegian and former Soviet Union experience after the Chernobyl accident, it is clear that a nuclear accident can entail decades of necessary management and rehabilitation of living conditions. The time period is dependent on a number of factors, e.g. amount of fallout, type of radionuclides, land use of contaminated area, number and density of people affected and available techniques and resources for implementing countermeasures. This paper discusses the management strategy implemented in Norway after the Chernobyl accident, the need for changing strategy over time and the important involvement of affected groups. Careful planning and reflections should be undertaken before actions are taken in the recovery phase, keeping in mind the possibility of decades with problems.


Dielectrophoresis of nanoparticles.
A numerical scheme based on the distributed Lagrange multiplier method (DLM) is used to study the motion of nano-sized particles of dielectric suspensions subjected to uniform and nonuniform electric fields. Particles are subjected to both electrostatic and hydrodynamic forces, as well as Brownian motion. The results of the simulations presented in this paper show that uniform electric fields the evolution of the particle structures depends on the ratio of electrostatic particle-particle interactions and Brownian forces. When this ratio is of the order of 100 or greater, particles form stable chains and columns, whereas when it is of the order of 10 or smaller the particle distribution is random. For the nonuniform electric field cases considered in this paper, the relative magnitude of Brownian forces is in the range such that it does not influence the eventual collection of particles by dielectrophoresis and the particular locations where the particles are collected. However, Brownian motion is observed to influence the transient particle trajectories. The deviation of the particle trajectories compared to those determined by the electrostatic and hydrodynamic forces alone is characterized by the ratio of Brownian and dielectrophoretic forces.


Does Schizophrenia in Offspring Increase the Risk of Developing Alzheimer's Dementia.
Prior studies have consistently found a higher risk of dementia in individuals with schizophrenia, but whether this is due to a common etiology between the disorders remains obscure. We wanted to elucidate this association by investigating whether schizophrenia in offspring increases the risk of Alzheimer's dementia. All individuals born between 1930 and 1953 were identified through national registers and followed from their 50th birthday until the date of Alzheimer's dementia, death or end of the study. Regressions were performed to evaluate the association between offspring with schizophrenia and Alzheimer's dementia. Individuals with offspring with schizophrenia did not have an increased risk of Alzheimer's dementia [incidence rate ratio (IRR), 0.97; 95% CI, 0.88-1.07] compared to individuals with offspring without psychiatric contact. This finding remained stable when evaluating early-onset (IRR, 1.10; 95% CI, 0.91-1.31) and late-onset Alzheimer's dementia (IRR, 0.92; 95% CI, 0.88-1.07). Similar findings were made for vascular and unspecified dementia. The finding of no familial coaggregation between schizophrenia and Alzheimer's dementia may suggest that no common etiology between the disorders exists. This may indicate that the excess risk of dementia in individuals with schizophrenia is a by-product of the higher rates of somatic comorbidity and adverse health risk factors among these individuals.


Review: Apolipoprotein E (Apo E) gene polymorphism and coronary heart disease in Asian populations.
Apolipoprotein E (Apo E) is a basic component of very low-density lipoprotein (VLDL) and high-density lipoprotein (HDL). It plays an important role in the clearance of cholesterol from circulation, and thereby slows down the process of atherosclerosis. Its 3 isoforms, E2, E3 and E4 are encoded by 3 alleles, e2, e3 and e4, respectively. E3 is the most common isoform in most populations in the world, while E2 is the least frequent isoform. A meta-analysis of several studies conducted on Asian populations revealed that carriers of e4 allele had 42% higher risk of coronary heart disease (CHD), while e2 allele had no significant association with this disease. Epidemiological studies performed in this region showed that E3 was the most prevalent isoform and most likely to be involved in CHD in Asia. Studies conducted in India indicated that individuals with e3/4 and e4/4 genotypes had considerably higher prevalence of dyslipidemia as compared to other genotypes, and hypertensive patients had high frequency of e4 allele. However, several other studies failed to show a relationship between ApoE gene polymorphism and CHD. Inadequate statistical power, low allele frequency, CHD phenotypes could be some of the possible reasons for conflicting results of some of these studies. This area of research is wide open and epidemiological studies with large sample size would be required to ascertain the relationships between ApoE gene polymorphism and CHD.


Mycobacterium tuberculosis lipoproteins directly regulate human memory CD4(+) T cell activation via Toll-like receptors 1 and 2.
The success of Mycobacterium tuberculosis as a pathogen relies on its ability to regulate the host immune response. M. tuberculosis can manipulate adaptive T cell responses indirectly by modulating antigen-presenting cell (APC) function or by directly interacting with T cells. Little is known about the role of M. tuberculosis molecules in direct regulation of T cell function. Using a biochemical approach, we identified lipoproteins LprG and LpqH as major molecules in M. tuberculosis lysate responsible for costimulation of primary human CD4(+) T cells. In the absence of APCs, activation of memory CD4(+) T cells with LprG or LpqH in combination with anti-CD3 antibody induces Th1 cytokine secretion and cellular proliferation. Lipoprotein-induced T cell costimulation was inhibited by blocking antibodies to Toll-like receptor 2 (TLR2) and TLR1, indicating that human CD4(+) T cells can use TLR2/TLR1 heterodimers to directly respond to M. tuberculosis products. M. tuberculosis lipoproteins induced NF-κB activation in CD4(+) T cells in the absence of TCR co-engagement. Thus, TLR2/TLR1 engagement alone by M. tuberculosis lipoprotein triggered intracellular signaling, but upregulation of cytokine production and proliferation required co-engagement of the TCR. In conclusion, our results demonstrate that M. tuberculosis lipoproteins LprG and LpqH participate in the regulation of adaptive immunity not only by inducing cytokine secretion and costimulatory molecules in innate immune cells but also through directly regulating the activation of memory T lymphocytes.


Cross-sectional evaluation of awareness of prevention of dental caries among general pediatricians in ghaziabad district, India.
Oral diseases are largely preventable and it is hoped that with the early exposure to oral health-care activities, the prevalence of oral diseases will be reduced in children and they would be more receptive to dental services. The present study evaluated the awareness of prevention of dental caries among pediatricians in Ghaziabad district, India. A cross-sectional survey was undertaken among the pediatricians in Ghaziabad district, India. Total subjects including in the survey were 88 pediatricians, through systemic random sampling. Both the gender was including Male-37.8% (35/88) and Female-62.2% (53/88). Pre-tested, structured and self administered questionnaire was used in the survey and data analysis was done by using 'SPSS' software version 16.0 (IBM, United States). Our study indicated that most of the pediatricians in Ghaziabad district had moderate knowledge 39.7% (35/88), followed by good knowledge 36.5% (32/88) and poor knowledge 23.8% (21/88) about dental caries. Practice guidelines and opinions of pediatricians in the survey were moderate 64.7% (57/88) in about more than half, followed by poor 23.8% (21/88) and followed by good 11.5% (10/88). The attitude for prevention of dental caries was positive in almost everybody 81.8% (72/88). The present survey concluded that pediatricians in Ghaziabad district, India had a good attitude and practices, but had moderate knowledge and lacked proper awareness about dental caries.


Acid, pepsin, and mucus secretion in patients with gastric and duodenal ulcer before and after colloidal bismuth subcitrate (De-Nol).
Basal and pentagastrin stimulated gastric secretion was measured in seven patients with duodenal, and six with gastric ulcers before and after four weeks' treatment with colloidal bismuth subcitrate (as De-Nol), one tablet four times a day. Each duodenal and all but one of the gastric ulcers healed. After De-Nol there were no significant changes in basal, or pentagastrin stimulated volume, acid output, or primary parietal component. There were marked decreases in basal (duodenal ulcer -25%; gastric ulcer -16%) and pentagastrin stimulated total pepsin outputs, (duodenal ulcer -42%, gastric ulcer -36%). There were insignificant decreases in basal output of mucus, but postpentagastrin stimulated mucus output was significantly inhibited (p less than 0.05) in patients with duodenal (-16%) and with gastric ulcer (-27%). The drop in gastric proteolysis after De-Nol is unlikely to be because of the healing of the ulcers and is more likely to be because of the drug. The ulcer healing efficacy of De-Nol may be related to this decline in the proteolytic action of gastric juice, but is unlikely to be because of a quantitative change in mucus, or in acid secretion.


The applicability of the Theory of Planned Behaviour in predicting adherence to ART among a South African sample.
We sought to determine the extent to which the Theory of Planned Behaviour (TPB) was applicable in predicting medication adherence among South Africans receiving antiretroviral therapy (ART). Regression analyses revealed that the linear combination of attitudes towards adherence, perceived behavioural control and perceived group norms explained 12 percent of the variance in intentions to adhere to ART. We also found a non-significant relationship between intentions to adhere to treatment and self-reported adherence. The results call into question the extent to which TPB is helpful in understanding a health-promoting behaviour such as medication adherence among South Africans receiving ART.


Quality of both parents' cognitive guidance and quantity of early childhood education: Influences on pre-mathematical development.
Only a handful of longitudinal studies have explored the effects of both parents in early parenthood on children's cognitive development, and no study has controlled for simultaneous early childhood education and care (ECEC) experiences. To examine the similarity of each parent's cognitive guidance and contribution to children's pre-mathematical outcomes across parent gender while controlling for amount of ECEC. A longitudinal study on 66 Finnish two-parent families and their children. Both parents' autonomy support and scaffolding behaviour were observed during play interactions with the child at 2;0. Children's numerical and spatial skills were tested at 3;0 and 4;0. Parental reports on amount of ECEC in months were collected at 2;0, 3;0, and 4;0. The two parenting constructs were conceptually similar across parent gender as demonstrated by fairly strong measurement equivalence. While mothers on average showed more scaffolding and equal amounts of autonomy support compared to fathers during play interaction, they displayed less variability in the former and more variability in the latter behaviour. The contribution of mothers' and fathers' cognitive guidance was more similar for children's numerical than spatial development. Both parents' education positively predicted autonomy support but not amount of ECEC exposure, which was positively related to numerical development. As expected, parents' education did not predict ECEC exposure, and child gender was not related to child outcomes. The findings are discussed in relation to measurement invariance and gender-neutral parenting in the context of early childhood.


Potential for large-scale CO2 removal via enhanced rock weathering with croplands.
Enhanced silicate rock weathering (ERW), deployable with croplands, has potential use for atmospheric carbon dioxide (CO2) removal (CDR), which is now necessary to mitigate anthropogenic climate change1. ERW also has possible co-benefits for improved food and soil security, and reduced ocean acidification2-4. Here we use an integrated performance modelling approach to make an initial techno-economic assessment for 2050, quantifying how CDR potential and costs vary among nations in relation to business-as-usual energy policies and policies consistent with limiting future warming to 2 degrees Celsius5. China, India, the USA and Brazil have great potential to help achieve average global CDR goals of 0.5 to 2 gigatonnes of carbon dioxide (CO2) per year with extraction costs of approximately US$80-180 per tonne of CO2. These goals and costs are robust, regardless of future energy policies. Deployment within existing croplands offers opportunities to align agriculture and climate policy. However, success will depend upon overcoming political and social inertia to develop regulatory and incentive frameworks. We discuss the challenges and opportunities of ERW deployment, including the potential for excess industrial silicate materials (basalt mine overburden, concrete, and iron and steel slag) to obviate the need for new mining, as well as uncertainties in soil weathering rates and land-ocean transfer of weathered products.


The effects of prenatal HIV exposure on language functioning in Kenyan children: establishing an evaluative framework.
HIV infection has been associated with impaired language development in prenatally exposed children. Although most of the burden of HIV occurs in sub-Saharan Africa, there have not been any comprehensive studies of HIV exposure on multiple aspects of language development using instruments appropriate for the population. We compared language development in children exposed to HIV in utero to community controls (N = 262, 8-30 months) in rural Kenya, using locally adapted and validated communicative development inventories. The mean score of the younger HIV-exposed uninfected infants (8-15 months) was not significantly below that of the controls; however older HIV-exposed uninfected children had significantly poorer language scores, with HIV positive children scoring more poorly than community controls, on several measures. Our preliminary data indicates that HIV infection is associated with impaired early language development, and that the methodology developed would be responsive to a more detailed investigation of the variability in outcome amongst children exposed to HIV, irrespective of their infection status.


High-throughput SNP discovery through deep resequencing of a reduced representation library to anchor and orient scaffolds in the soybean whole genome sequence.
The Soybean Consensus Map 4.0 facilitated the anchoring of 95.6% of the soybean whole genome sequence developed by the Joint Genome Institute, Department of Energy, but its marker density was only sufficient to properly orient 66% of the sequence scaffolds. The discovery and genetic mapping of more single nucleotide polymorphism (SNP) markers were needed to anchor and orient the remaining genome sequence. To that end, next generation sequencing and high-throughput genotyping were combined to obtain a much higher resolution genetic map that could be used to anchor and orient most of the remaining sequence and to help validate the integrity of the existing scaffold builds. A total of 7,108 to 25,047 predicted SNPs were discovered using a reduced representation library that was subsequently sequenced by the Illumina sequence-by-synthesis method on the clonal single molecule array platform. Using multiple SNP prediction methods, the validation rate of these SNPs ranged from 79% to 92.5%. A high resolution genetic map using 444 recombinant inbred lines was created with 1,790 SNP markers. Of the 1,790 mapped SNP markers, 1,240 markers had been selectively chosen to target existing unanchored or un-oriented sequence scaffolds, thereby increasing the amount of anchored sequence to 97%. We have demonstrated how next generation sequencing was combined with high-throughput SNP detection assays to quickly discover large numbers of SNPs. Those SNPs were then used to create a high resolution genetic map that assisted in the assembly of scaffolds from the 8x whole genome shotgun sequences into pseudomolecules corresponding to chromosomes of the organism.


[Identification of genes that predict lymph node metastasis in colorectal cancer cases].
Currently, Endoscopic mucosal resection (EMR) and laparoscopic surgery with colorectal cancer (CRC) and has been expanding rapidly. In handling of colon cancer, adaptation of EMR is determined by the depth of tumor invasion. It is important to identify genes to predict lymph node metastasis in early CRC tumors precisely in a reproducible fashion to determine the adaptation of EMR treatment. We performed the comprehensive analysis of gene expression and genomic copy number simultaneously in CRC primary tumors to identify the bona-fide indicator of lymph node metastasis. We collected cancer cells specifically by Laser Microdissection (LMD) on 157 cases of primary colorectal cancer, and performed oligo microarrays for gene expression (GE) and aCGH for copy number aberration. As for candidate genes to be associated with lymph node metastasis, we examined reprodicibility by quantitative RT-PCR using cDNA created from the RNA extracted from 172 cases of CRC. As for the association of lymph node metastasis, we found that 240 genes and 54 genes by aCGH and by oligo GE microarray, respectively. According to database of those two arrays, 501 genes were significantly correlated (correlation coefficient > 0.7) with each other, and we found that 11 out of 501 genes were identified as lymph node metastasis related genes with copy number alteration. Of these 11 genes, we focused on PCM1, MTUS1, ASAH1 on 8p22. Then, we confirmed that the decreased expression and genomic deletion of MTUS1 were observed in lymph node positive cases (p = 0.0195) in another subset of 172 cases of CRC. To measure the expression of MTUS1 of the tumor by PCR, we can predict the presence of lymph node metastasis. We expected that the loss of MTUS1 should be an important marker in determining the adaptation of endoscopic resection.


Association of early onset periodontitis microbiota with aspartate aminotransferase activity in gingival crevicular fluid.
The objective of this study was to determine the relationship between the activity of the enzyme aspartate aminotransferase (AST) in gingival crevicular fluid (GCF) using the colorimetric PerioGard (PTM) test and the subgingival microflora in early onset periodontitis lesions. The study population consisted of 25 otherwise healthy individuals exhibiting early onset periodontitis (EOP). In each patient four experimental sites were identified comprising one deep periodontal pocket (PD >5 mm) randomly chosen in each quadrant. Bacterial samples were obtained from the experimental sites, consecutively cultured anaerobically and in 10% CO(2) using selective and nonselective media. Isolates were characterized to species level by conventional biochemical tests and various identification kits. Clinical measurements as well as AST activity, assessed either as positive or negative using the PTM, were recorded at the same sites. Sixty-two sites exhibited AST positive and 38 AST negative activity. Analysis of bacterial counts using the ANOVA (Mann Whitney U-test) showed that Streptococcus intermedius, Peptostreptococcus micros, Campylobacter concisus, Bacteroides forsythus, Camplobacter gracilis, Campylobacter rectus and Selenomonas sputigena were significantly higher in sites with AST-positive activity. The odds ratio of having high prevalence of S. intermedius, P. micros, C. concisus, B. forsythus, C. gracilis, C. rectus and S. sputigena in the presence of a positive AST site was very high (range: 3.5-17.0). Streptococcus sanguis, Actinomyces naeslundii, Gemella morbillorum, Capnocytophaga gingivalis, Veillonella parvula, Fusobacterium varium, Eubacterium lentum and Prevotella oralis were detected in significantly higher proportions in sites with AST negative activity and manifested a negative odds ratio in the presence of AST positive sites. The logistic regression analysis revealed that smoking and bleeding upon probing showed a significant association with AST activity, while plaque and suppuration were not found to be significant predictors of AST activity. The co-infection of Porphyromonas gingivalis, B. forsythus and P. micros, or P. gingivalis, B. forsythus and C. rectus were found to be significantly associated with the AST activity (p<0.001). AST positive sites revealed significantly higher occurrence of co-infections by P. gingivalis, B. forsythus, S. sputigena or by P. gingivalis, B. forsythus, S. intermedius than AST negative sites (p<0.001). P. gingivalis, B. forsythus, A. naeslundii co-infection was found significantly higher in the AST negative sites (p<0.001). The present study found a high level of agreement between the presence of putative periodontal pathogens and positive AST scores at periodontal sites that clinically were considered to be potentially disease active. Prospective studies should be performed to confirm the findings.


Prevention of anastrozole induced bone loss with monthly oral ibandronate: Final 5 year results from the ARIBON trial.
The ARIBON trial is a double blind, randomised, placebo controlled study designed to evaluate the impact of ibandronate on bone mineral density (BMD) in women taking anastrozole for adjuvant treatment of breast cancer. 131 postmenopausal women with early breast cancer were recruited to the study. Of these, 13 had osteoporosis, 50 osteopenia and 68 normal BMD. Patients with osteoporosis at baseline were treated with monthly oral ibandronate 150 mg for 5 years; osteopenic patients were randomised to receive either ibandronate or placebo for two years and offered open label ibandronate depending upon the results of their 2-year BMD result. Of the 20 patients with osteopenia who were randomised to ibandronate and evaluable at the 2 year visit, 17/20 were not offered a bisphosphonate and the improvements in BMD accrued during the first 2 years were lost both at the LS (-3.21%) and TH (-5.0%). Of the 16 patients randomised to placebo 8/16 with high rates of bone loss during years 0-2 received ibandronate over the next 3 years with improvements in BMD of +5.01 and +1.19 at the LS and TH respectively. The 8 patients who were not offered a bisphosphonate experienced relatively little change in BMD throughout the 5 years of the study (LS +0.15%, TH -2.72%). BMD increased steadily in the 9/13 patients initially identified as having osteoporosis (LS +9.65%, TH +2.72%). Monthly oral ibandronate provides an option to clinicians considering use of a bisphosphonate to prevent bone loss during aromatase inhibitor therapy.


Xenopus laevis as a Bioindicator of Endocrine Disruptors in the Region of Central Chile.
One of the direct causes of biodiversity loss is environmental pollution resulting from the use of chemicals. Different kinds of chemicals, such as persistent organic pollutants and some heavy metals, can be endocrine disruptors, which act at low doses over a long period of time and have a negative effect on the reproductive and thyroid system in vertebrates worldwide. Research on the effects of endocrine disruptors and the use of bioindicators in neotropical ecosystems where pressure on biodiversity is high is scarce. In Chile, although endocrine disruptors have been detected at different concentrations in the environments of some ecosystems, few studies have been performed on their biological effects in the field. In this work, Xenopus laevis (African clawed frog), an introduced species, is used as a bioindicator for the presence of endocrine disruptors in aquatic systems with different degrees of contamination in a Mediterranean zone in central Chile. For the first time for Chile, alterations are described that can be linked to exposure to endocrine disruptors, such as vitellogenin induction, decreased testosterone in male frogs, and histological changes in gonads. Dioxin-like and oestrogenic activity was detected in sediments at locations where it seem to be related to alterations found in the frogs. In addition, an analysis of land use/cover use revealed that urban soil was the best model to explain the variations in frog health indicators. This study points to the usefulness of an invasive species as a bioindicator for the presence of endocrine-disruptive chemicals.


Rhamnocitrin decreases fibrosis of ovarian granulosa cells by regulating the activation of the PPARγ/NF-κB/TGF-β1/Smad2/3 signaling pathway mediated by Wisp2.
Polycystic ovary syndrome (PCOS) is the most common cause of anovulatory infertility in women. Rhamnocitrin (Rha) has anti-inflammatory and antioxidant actions. The WNT1-inducible-signaling pathway protein 2 (Wisp2) and nuclear factor (NF)-κB are involved in fibrosis in many diseases. We aimed to elucidate the role of Rha in fibrosis of PCOS and the underlying mechanisms. Dehydroepiandrosterone (DHEA)-incubated ovarian granulosa KGN cells were treated by Rha. Cell proliferation was detected with cell counting kit-8 (CCK-8) and 5-ethynyul-2'-deoxyuridine (EdU) staining. The levels of Wisp2 and transforming growth factor-β1 (TGF-β1) in supernatant were measured by enzyme-linked immunosorbent assay (ELISA). We observed α-smooth muscle actin (α-SMA) protein by immunofluorescence (IF). The levels of fibrosis factors were determined using Western blot. We observed p65 nuclear translocation with confocal microscopy. We used Wisp2 overexpression and knockdown in cells treated with DHEA or Rha to validate Wisp2 function. Interaction between Wisp2 and NF-κB, as well as Wisp2 and PPARγ, were assessed by co-immunoprecipitation assay, luciferase reporter assay and chromatin immunoprecipitation (ChIP). The results showed that Rha elevated the reduced proliferation of DHEA-treated cells. In addition, Rha reversed the decreased Wisp2 and the increased TGF-β1 in supernatant. The proteins CTGF, α-SMA, Collagen I, TGF-β1, p-Smad2, and p-Smad3 were up-regulated while Wisp2, Sirt1, and PPARγ were down-regulated by DHEA treatment, which were reversed by Rha. Meanwhile, DHEA up-regulated p-IKBa and p-p65 and promoted p65 nuclear translocation, which were inhibited by Rha. These effects of Rha were antagonized by Wisp2 knockdown and were mimicked by Wisp2 overexpression. We confirmed the protein interaction between Wisp2 and NF-κB, along with Wisp2 and PPARγ. Wisp2-mediated PPARγ/NF-κB/TGF-β1/Smad2/3 signaling contributes to Rha-improved ovarian granulosa cells fibrosis, suggesting Rha as a novel agent for the treatment of PCOS.


Duration of intrathecal labor analgesia: early versus advanced labor.
Early first-stage labor pain is primarily visceral in origin. Increasing pain intensity and transition to somatic nociceptive input characterizes late first- and second-stage labor pain. The effect of this change in nociceptive input on the duration of intrathecal labor analgesia has not been well studied. This prospective cohort observational study compares the duration of intrathecal labor analgesia after intrathecal injections made in early labor (3- to 5-cm cervical dilation) and those made in more advanced labor (7- to 10-cm cervical dilation). Forty-one parturients (18 in early labor and 23 in advanced labor) received intrathecal sufentanil (10 micrograms) and bupivacaine (2.5 mg) as part of a combined spinal-epidural technique. Patients rated their pain using a 0-10 verbal pain scale prior to intrathecal injection and every 20 min thereafter. Duration of analgesia was defined as the lesser of time until the pain score exceeded 5 or until a request for supplemental epidural analgesia was made. The duration of spinal analgesia was significantly less when intrathecal injection was made in advanced labor (120 +/- 26 min) compared with early labor (163 +/- 57 min, P < 0.01). We conclude that cervical dilation and stage of labor significantly impact the effective duration of intrathecal sufentanil/ bupivacaine labor analgesia.


Diagnostic performance of anti-Zika virus IgM, IgAM and IgG ELISAs during co-circulation of Zika, dengue, and chikungunya viruses in Brazil and Venezuela.
Serological diagnosis of Zika virus (ZIKV) infection is challenging because of the antibody cross-reactivity among flaviviruses. At the same time, the role of Nucleic Acid Testing (NAT) is limited by the low proportion of symptomatic infections and the low average viral load. Here, we compared the diagnostic performance of commercially available IgM, IgAM, and IgG ELISAs in sequential samples during the ZIKV and chikungunya (CHIKV) epidemics and co-circulation of dengue virus (DENV) in Brazil and Venezuela. Acute (day of illness 1-5) and follow-up (day of illness ≥ 6) blood samples were collected from nine hundred and seven symptomatic patients enrolled in a prospective multicenter study between June 2012 and August 2016. Acute samples were tested by RT-PCR for ZIKV, DENV, and CHIKV. Acute and follow-up samples were tested for IgM, IgAM, and IgG antibodies to ZIKV using commercially available ELISAs. Among follow-up samples with a RT-PCR confirmed ZIKV infection, anti-ZIKV IgAM sensitivity was 93.5% (43/46), while IgM and IgG exhibited sensitivities of 30.3% (10/33) and 72% (18/25), respectively. An additional 24% (26/109) of ZIKV infections were detected via IgAM seroconversion in ZIKV/DENV/CHIKV RT-PCR negative patients. The specificity of anti-ZIKV IgM was estimated at 93% and that of IgAM at 85%. Our findings exemplify the challenges of the assessment of test performance for ZIKV serological tests in the real-world setting, during co-circulation of DENV, ZIKV, and CHIKV. However, we can also demonstrate that the IgAM immunoassay exhibits superior sensitivity to detect ZIKV RT-PCR confirmed infections compared to IgG and IgM immunoassays. The IgAM assay also proves to be promising for detection of anti-ZIKV seroconversions in sequential samples, both in ZIKV PCR-positive as well as PCR-negative patients, making this a candidate assay for serological monitoring of pregnant women in future ZIKV outbreaks.


Inflammatory and hemostatic response to cardiopulmonary bypass in pediatric population: feasibility of seriological testing of multiple biomarkers.
Perioperative myocardial and cerebral damages are the major determinants of postoperative morbidity and mortality in pediatric cardiac surgery. Cardiopulmonary bypass (CPB) causes alterations in the levels of biomarkers related to inflammation, tissue damage, and other tissue pathologies. Early and accurate evaluation of inflammation and tissue damage would therefore be clinically useful. Our objective is to assess the suitability of using Multi-Analyte Profiling (MAP) (Rules Based Medicine, Austin, TX, USA) in pediatric cardiac surgery as a potential surrogate marker of clinical outcome. MAP technology platform allowed us to analyze 90 different biomarkers using only 100µL of plasma to detect any changes in the levels of 90 biomarkers. Plasma samples (100µL) were collected at five different time points: 1. before midline incision; 2. on CPB for 3-5min; 3. at the end of CPB; 4. 1h after CPB; and 5. 24h after CPB. After removing the outliers, the average and standard deviation of the values obtained from the 10 patients were calculated for each time point. The average values of each biomarker at each time point were then compared to each other and to the baseline. The pilot protocol included 10 patients (ages from 3 months to 4 years old) with similar Jenkins risks stratifications who underwent nonpulsatile CPB. We detected changes in the levels of 90 biomarkers. Biomarkers were assessed in groups. Myeloperoxidase (MPO) and pregnancy-associated plasma protein A (PAPP-A) were the earliest markers to rise with 49- and 18-fold increases 3-5min after the onset of CPB, respectively. The most striking increase was noted in the heart-type fatty acid-binding protein (FABP) levels. FABP increased 25, 193, 151, and 4-fold at time points 2, 3, 4, and 5, respectively. Surges in the novel markers of injury were followed by the markers of inflammation (i.e., C-reactive protein, interleukins) peaking at 24h after CPB. This pilot study shows that it is possible to measure 90 different biomarkers using only a very small sample of plasma to evaluate the effects of CPB. Novel markers of tissue injury (FABP, PAPP-A, or MPO) are the earliest markers to rise. Serial monitoring of multiple biomarkers may help to predict and improve outcomes after pediatric cardiac surgery.


Effects of a healthy Nordic diet on gene expression changes in peripheral blood mononuclear cells in response to an oral glucose tolerance test in subjects with metabolic syndrome: a SYSDIET sub-study.
Diet has a great impact on the risk of developing features of metabolic syndrome (MetS), type 2 diabetes mellitus (T2DM), and cardiovascular diseases (CVD). We evaluated whether a long-term healthy Nordic diet (ND) can modify the expression of inflammation and lipid metabolism-related genes in peripheral blood mononuclear cells (PBMCs) during a 2-h oral glucose tolerance test (OGTT) in individuals with MetS. A Nordic multicenter randomized dietary study included subjects (n = 213) with MetS, randomized to a ND group or a control diet (CD) group applying an isocaloric study protocol. In this sub-study, we included subjects (n = 89) from three Nordic centers: Kuopio (n = 26), Lund (n = 30), and Oulu (n = 33) with a maximum weight change of ±4 kg, high-sensitivity C-reactive protein concentration ≤10 mg L(-1), and baseline body mass index <39 kg m(-2). PBMCs were isolated, and the mRNA gene expression analysis was measured by quantitative real-time polymerase chain reaction (qPCR). We analyzed the mRNA expression changes of 44 genes before and after a 2hOGTT at the beginning and the end of the intervention. The healthy ND significantly down-regulated the expression of toll-like receptor 4 (TLR4), interleukin 18 (IL18), and thrombospondin receptor (CD36) mRNA transcripts and significantly up-regulated the expression of peroxisome proliferator-activated receptor delta (PPARD) mRNA transcript after the 2hOGTT compared to the CD. A healthy ND is able to modify the gene expression in PBMCs after a 2hOGTT. However, more studies are needed to clarify the biological and clinical relevance of these findings.


The structures of complexes between polyethylene imine and sodium dodecyl sulfate in D2O: a scattering study.
The association between a highly branched polyelectrolyte with ionizable groups, polyethylene imine (PEI), and an anionic surfactant, sodium dodecyl sulfate (SDS), has been investigated at two pH values, using small-angle neutron and light scattering. The scattering data allow us to obtain a detailed picture of the association structures formed. Small-angle neutron scattering (SANS) measurements in solutions containing highly charged PEI at low pH and low SDS concentrations indicate the presence of disklike aggregates. The aggregates change to a more complex three-dimensional structure with increasing surfactant concentration. One pronounced feature in the scattering curves is the presence of a Bragg-like peak at high q-values observed at a surfactant concentration of 4.2 mM and above. This scattering feature is attributed to the formation of a common well-ordered PEI/SDS structure, in analogue to what has been reported for other polyelectrolyte-surfactant systems. Precipitation occurred at the charge neutralization point, and X-ray diffraction measurements on the precipitate confirmed the existence of an ordered structure within the PEI/SDS aggregates, which was identified as a lamellar internal organization. Polyethylene imine has a low charge density in alkaline solutions. At pH 10.1 and under conditions where the surfactant was contrast matched, the SANS scattering curves showed only small changes with increasing surfactant concentration. This suggests that the polymer acts as a template onto which the surfactant molecules aggregate. Data from both static light scattering and SANS recorded under conditions where SDS and to a lower degree PEI contribute to the scattering were found to be consistent with a structure of stacked elliptic bilayers. These structures increased in size and became more compact as the surfactant concentration was increased up to the charge neutralization point.


Simplest relationship between local field potential and intracellular signals in layered neural tissue.
The relationship between the extracellularly measured electric field potential resulting from synaptic activity in an ensemble of neurons and intracellular signals in these neurons is an important but still open question. Based on a model neuron with a cylindrical dendrite and lumped soma, we derive a formula that substantiates a proportionality between the local field potential and the total somatic transmembrane current that emerges from the difference between the somatic and dendritic membrane potentials. The formula is tested by intra- and extracellular recordings of evoked synaptic responses in hippocampal slices. Additionally, the contribution of different membrane currents to the field potential is demonstrated in a two-population mean-field model. Our formalism, which allows for a simple estimation of unknown dendritic currents directly from somatic measurements, provides an interpretation of the local field potential in terms of intracellularly measurable synaptic signals. It is also applicable to the study of cortical activity using two-compartment neuronal population models.


Improved multilocus sequence typing of Burkholderia pseudomallei and closely related species.
The Burkholderiapseudomallei multilocus sequence typing (MLST) database (http://pubmlst.org/bpseudomallei/) contains the largest global sequence repository for B. pseudomallei and its closest genetic relatives. Using conventional MLST and in silico MLST data derived from publicly available whole-genome sequences, we first defined the phylogenetic relatedness of B. pseudomallei and its nearest neighbours. Based on this analysis, we propose that the recently described B. pseudomallei complex (Bpc) should be expanded to encompass B. pseudomallei, Burkholderiahumptydooensis (proposed), Burkholderiamallei, Burkholderiaoklahomensis, Burkholderiathailandensis and three unassigned Burkholderia Clades A, B and C (represented by type strains BDU 5, BDU 8 and MSMB0265, respectively). Of note, the MLST narK locus is present in all Bpc species but is missing in all other Burkholderia spp., including all Burkholderiacepacia complex species, with the exception of most Burkholderiaubonensis strains, which contain narK but encode genetically distinct sequences. The presence of narK is thus indicative of a Bpc strain. Next, we revisited in silico the performance of the existing MLST primers, which prompted redesign of primers targeting the gmhD, lepA, lipA, narK and ndh loci to encompass genetic diversity among Bpc strains and to address amplification/sequencing issues. We show in silico and in vitro that the redesigned primers yield good-quality amplification and sequencing results for the gmhD, lepA, lipA, narK and ndh loci in Bpc species. These primers provide an alternative for amplification and sequencing of MLST loci in Bpc species in cases when poor-quality amplification or sequencing data are obtained using the original MLST primers.


Public health practitioners' views of the 'Making Every Contact Count' initiative and standards for its evaluation.
National Health Service England encourages staff to use everyday interactions with patients to discuss healthy lifestyle changes as part of the 'Making Every Contact Count' (MECC) approach. Although healthcare, government and public health organisations are now expected to adopt this approach, evidence is lacking about how MECC is currently implemented in practice. This study explored the views and experiences of those involved in designing, delivering and evaluating MECC. We conducted a qualitative study using semi-structured interviews with 13 public health practitioners with a range of roles in implementing MECC across England. Interviews were conducted via telephone, transcribed verbatim and analysed using an inductive thematic approach. Four key themes emerged identifying factors accounting for variations in MECC implementation: (i) 'design, quality and breadth of training', (ii) 'outcomes attended to and measured', (iii) 'engagement levels of trainees and trainers' and (iv) 'system-level influences'. MECC is considered a valuable public health approach but because organisations interpret MECC differently, staff training varies in nature. Practitioners believe that implementation can be improved, and an evidence-base underpinning MECC developed, by sharing experiences more widely, introducing standardization to staff training and finding better methods for assessing meaningful outcomes.


Surface adsorption and self-assembly of Cu(II) ions on TEMPO-oxidized cellulose nanofibers in aqueous media.
TEMPO-mediated oxidized cellulose nanofibers (TOCNFs) have shown potential in the bioremediation of metal ions from contaminated water due to their interaction with positively charged metal ions via electrostatic interactions involving surface carboxyl groups. Copper is one of the most common pollutants in industrial effluents and is thus the target metal in the current study. The specific surface adsorption of Cu(II) was similar for TOCNFs with different degrees of functionalization and directly impacted the zeta potential. SEM imaging of the TOCNF after Cu(II) adsorption revealed interesting nanostructured clusters that were attributable to Cu(II) ions first being adsorbed by carboxylate groups on the TOCNF and subsequently being reduced and self-assembled to Cu(0) nanoparticles (NPs) or copper oxide NPs by microprecipitation. TOCNF turned superhydrophilic and resulted in faster water filtration after copper adsorption due to the stronger polarity of the copper ions or the self-assembled Cu(0) NPs creating voids or highly water-permeable channels at the interface between the interconnected TEMPO-oxidized nanofibers. Thus, the adsorption of Cu(II) ions and self-assembly into the Cu NPs on TOCNF favors a faster water purification process and provides a viable route to reuse/recycle TOCNFs studded with Cu nanoparticles as biocidal materials.


Femtosecond laser pumped by high-brightness coherent polarization locked diodes.
We demonstrate, for the first time to our knowledge, the use of a coherent polarization locked diode as the high-brightness pump source for a femtosecond laser. Four diode emitters are coherently locked to produce more than 5 W linearly polarized, narrow linewidth, and single-lobed pump beam. This gives >10× brightness improvement over the conventional diode array. The diode beam is then used to pump a Yb:KYW laser to obtain 2 W output with 57% slope efficiency in cw laser operation. By using a saturable absorber mirror, we achieved cw mode-locking operation with a 177 fs pulse width at an average power of 0.55 W.


Regional distribution of mutations of the ATP7B gene in patients with Wilson disease: impact on genetic testing.
Wilson disease is an autosomal recessive inherited disorder of copper metabolism. The Wilson disease gene codes for a copper transporting P-type ATPase (ATP7B). Molecular genetic analysis reveals at least 300 distinct mutations. While most reported mutations occur in single families, a few are more common. The most common mutation in patients from Central, Eastern, and Northern Europe is the point mutation H1069Q (exon 14). About 50-80% of Wilson disease (WD) patients from these countries carry at least one allele with this mutation with an allele frequency ranging between 30 and 70%. Other common mutations in Central and Eastern Europe are located on exon 8 (2299insC, G710S), exon 15 (3400delC) and exon 13 (R969Q). The allele frequency of these mutations is lower than 10%. In Mediterranean countries there is a wide range of mutations, the frequency of each of them varies considerably from country to country. In Sardinia, a unique deletion in the 5' UTR (-441/-427 del) is very frequent. In mainland Spain the missense mutation M645R in exon 6 is particularly common. Data from non-European countries are scarce. Most data from Asia are from Far Eastern areas (China, South Korea and Japan) where the R778L missense mutation in exon 8 is found with an allele frequency of 14-49%. In summary, given the constant improvement of analytic tools genetic testing will become an integral part for the diagnosis of WD. Knowledge of the differences in the worldwide distribution of particular mutations will help to design shortcuts for genetic diagnosis of WD.


Effects of palm oil tocotrienol-rich fraction on biochemical and morphological alterations of liver in fenitrothion-treated rats.
Indiscriminate application of organophosphate (OP) pesticides has led to environmental pollution and severe health problems. The aim of the present study was to evaluate the effect of palm oil tocotrienol-rich fraction (TRF) on biochemical and morphological changes of the liver in rats treated with fenitrothion (FNT), a type of OP pesticide. A total of 28 male Sprague-Dawley rats were divided into four groups; control group, TRF-supplemented group, FNT-treated group and TRF+FNT group. TRF (200 mg/kg) was supplemented 30 minutes prior to FNT (20 mg/kg) administration, both orally for 28 consecutive days. Following 28 days of treatment, plasma biochemical changes and liver morphology were evaluated. The body and absolute liver weights were significantly elevated in TRF+FNT group compared to FNT group. TRF administration significantly decreased the total protein level and restored the activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in TRF + FNT group. In contrast, total bilirubin level, γ-glutamyltranferase (GGT) and cholinesterase activity in TRF + FNT group did not significantly differ from FNT group. Administration of TRF also prevented FNT-induced morphological changes of liver as observed by electron microscope. In conclusion, TRF supplementation showed potential protective effect towards biochemical and ultrastructural changes in liver induced by FNT.


Neuron-derived orphan receptor 1 transduces survival signals in neuronal cells in response to hypoxia-induced apoptotic insults.
OBJECT Hypoxia can induce cell death or trigger adaptive mechanisms to guarantee cell survival. Neuron-derived orphan receptor 1 (NOR-1) works as an early-response protein in response to a variety of environmental stresses. In this study, the authors evaluated the roles of NOR-1 in hypoxia-induced neuronal insults. METHODS Neuro-2a cells were exposed to oxygen/glucose deprivation (OGD). Cell viability, cell morphology, cas-pase-3 activity, DNA fragmentation, and cell apoptosis were assayed to determine the mechanisms of OGD-induced neuronal insults. RNA and protein analyses were carried out to evaluate the effects of OGD on expressions of NOR-1, cAMP response element-binding (CREB), and cellular inhibitor of apoptosis protein 2 (cIAP2) genes. Translations of these gene expressions were knocked down using RNA interference. Mice subjected to traumatic brain injury (TBI) and NOR-1 was immunodetected. RESULTS Exposure of neuro-2a cells to OGD decreased cell viability in a time-dependent manner. Additionally, OGD led to cell shrinkage, DNA fragmentation, and cell apoptosis. In parallel, treatment of neuro-2a cells with OGD time dependently increased cellular NOR-1 mRNA and protein expressions. Interestingly, administration of TBI also augmented NOR-1 levels in the impacted regions of mice. As to the mechanism, exposure to OGD increased nuclear levels of the transcription factor CREB protein. Downregulating CREB expression using RNA interference simultaneously inhibited OGD-induced NOR-1 mRNA expression. Also, levels of cIAP2 mRNA and protein in neuro-2a cells were augmented by OGD. After reducing cIAP2 translation, OGD-induced cell death was reduced. Sequentially, application of NOR-1 small interfering RNA to neuro-2a cells significantly inhibited OGD-induced cIAP2 mRNA expression and concurrently alleviated hypoxia-induced alterations in cell viability, caspase-3 activation, DNA damage, and cell apoptosis. CONCLUSIONS This study shows that NOR-1 can transduce survival signals in neuronal cells responsible for hypoxiainduced apoptotic insults through activation of a CREB/cIAP2-dependent mechanism.


Pharmacokinetics of buprenorphine following intravenous and intramuscular administration in male rhesus macaques (Macaca mulatta).
This study reports the pharmacokinetics of buprenorphine in conscious rhesus macaques (Macaca mulatta) after intravenous (i.v.) and intramuscular (i.m.) administration. Four healthy, opioid-naïve, socially housed, adult male macaques were used. Buprenorphine (0.03 mg/kg) was administered intravenously as a bolus or intramuscularly on separate occasions. Blood samples were collected prior to, and up to 24 h, postadministration. Serum buprenorphine concentrations were analyzed with liquid chromatography-mass spectrometry. Noncompartmental pharmacokinetic analysis was performed with commercially available software. Mean residence time in the i.v. study as compared to the i.m. study was 177 (159-189) vs. 185 (174-214) min, respectively [median (range)]. In the i.v. study, concentration back-extrapolated to time zero was found to be 33.0 (16.8-57.0) ng/mL [median (range)]. On the other hand, the maximum serum concentration found in the i.m. study was 11.8 (6.30-14.8) ng/mL [median (range)]. Rhesus macaques maintained concentrations >0.10 ng/mL for over 24 h in the i.v. study and over 12 h in the i.m. study. Bioavailability was found to be 68.1 (59.3-71.2)% [median (range)]. No significant adverse effects were observed in the monkeys at the 0.03 mg/kg dose of buprenorphine during either study.


Kinetics of proliferation of rat aortic smooth muscle cells in Goldblatt one-kidney, one-clip hypertension.
Wistar female rats were made hypertensive by applying a silver clip to the left renal artery and removing the right kidney. In aortas, the proliferation fraction of smooth muscle cells, DNA synthesis and wet weight have been correlated with the blood pressure increase subsequent to the operation. A wave of proliferation of smooth muscle cells in the aortic media is triggered immediately after the highest increased rate of blood pressure. When blood pressure stabilizes at high values, the metabolism of nucleic acid within the aortic media resumes its normal level but the arterial changes previously established persist. The sequence of pathological events responsible for these changes could be: increment of blood pressure; increase in wall stress; proliferation of smooth muscle cells; thickening of arterial wall; correction of the wall stress; end of proliferation. The consequence of this early proliferation of aortic smooth muscle cells is not clear but it is probably one of the mechanisms through which high blood pressure is sustained. It can also participate in atherogenesis, being in this way one of the bases of the well-known relationship between hypertension and atherosclerosis.


Normative Data for a Comprehensive Neuropsychological Test Battery used in the Assessment of Sports-Related Concussion.
The use of normative data is a hallmark of the neuropsychological assessment process. Within the context of sports-related concussion, utilizing normative data is especially essential when individualized baseline data are unavailable for comparison. The primary purpose of this study was to establish normative data for a comprehensive neuropsychological test battery used in the assessment of sports-related concussion. A secondary aim was to provide normative data for pertinent demographic variables relevant to the assessment of college athletes, including sex, previous head injuries (PHI), and history of attention deficit hyperactivity disorder (ADHD)/learning disability (LD). Participants included male and female college athletes (N = 794) who were involved in a concussion management program at an NCAA Division I university between 2002 and 2015. Athletes were administered a comprehensive neuropsychological test battery at baseline designed to assess the following cognitive domains: learning and memory, attention and concentration, processing speed, and executive functioning. The test battery primarily comprises paper-and-pencil measures. Normative data are presented for the overall athlete sample. Additional sub-norms are then provided for specified demographic populations (i.e., sex, PHIs, and history of ADHD/LD). Findings indicate that there are mild cognitive differences between men and women, as well as between those athletes with and without a history of ADHD/LD. Given these findings, additional norms are provided for men and women with and without a history of ADHD/LD. In the absence of baseline testing, the normative data presented here can be used clinically to assess athletes' cognitive functioning post-concussion.


Clinical decision making regarding suicide risk: Effect of patient and clinician age.
To ascertain how patient age influences suicide risk assessment, clinicians (N = 262) read an ambiguous vignette about Bill (aged either 39 or 79 years old) and subsequently rated Bill's suicide risk and hospitalization needs. Suicide-risk ratings varied greatly and young clinicians rated Bill's suicide risk and hospitalization needs higher when he was elderly (79 years old); whereas, older clinicians rated Bill's suicide risk and need for hospitalization higher when he was younger (39 years old). The interaction between patient and clinician age may reflect a "similarity" bias, such that clinicians perceive those who are different (i.e., younger or older) to be at elevated risk.


[Academic achievement, engagement and burnout among first year medical students].
Stress may affect the sense of wellbeing and academic achievement of university students. To assess the relationship of academic engagement and burnout with academic achievement among first year medical students. The Utrecht Work Engagement Scale-Student and Maslach Burnout Inventory Student Survey (MBI-SS) were applied to 277 first year medical students of four universities. Their results were correlated with the grades obtained in the different courses. Moderately high engagement and low burnout levels were detected. There was a high level of satisfaction with studies and a moderate exhaustion level. Academic achievement was associated with the degree of engagement with studies but not with burnout. Conglomerate analysis detected a group of students with high levels of wellbeing, characterized by high levels of academic engagement and low burnout. Other group had moderate levels of engagement and lack of personal fulfilment. Other group, identified as extenuated, had high levels of personal exhaustion and depersonalization. Finally the disassociated group had a low academic engagement, low emotional exhaustion, high levels of depersonalization and lack of personal fulfillment. Academic achievement is associated with the level of engagement with studies but not with burnout.


RSV vaccine use--the missing data.
Respiratory syncytial virus (RSV) infection is the most important cause of hospitalization in infants and is one of the leading global causes of infant mortality and as such its prevention through vaccination is a public health priority. While essential for the successful implementation of vaccine programs, there remains a paucity of data on the epidemiology of the virus in different settings and age groups and limited knowledge about virus transmission and the health-care costs of the disease. Such data are now needed to populate health economic models and to inform optimal approaches to disease control through vaccination.


"Embracing the present and fearing the future": the meaning of being an oldest old woman in a rural area.
In Western countries, the number of older people receiving home nursing care is increasing, and in rural areas they are at additional risk because of the distance between people and health care facilities. The aim of this study was therefore to illuminate the meaning of being an oldest old woman living alone in a rural area and receiving home nursing care. A sample of 11 oldest old women living in rural areas in the middle of Norway was chosen for this study. Narrative interviews were conducted, and the data were analyzed using the phenomenological hermeneutic method. After a naïve reading and a structural analysis of the text, we identified four themes: being satisfied with life, being thankful, feeling vulnerable, and feeling secure. The comprehensive understanding implied that being an oldest old woman living alone in a rural area meant living in the intersection between embracing the present in solitude and fearing the future with additional declining health. Living in this complex situation meant to enjoy the present, but still fear the future, as the oldest old women knew their present life situations were limited. This challenging emotional situation meant using their inner strength by trying to be optimistic and seeing opportunities in present life, even if losses were many and extensive. By using their inner strength in facing losses and declining health, the oldest old women managed to appreciate aloneness as solitude, and find new meaning in life.


Elimination of the transient ipsilateral retinotectal projection is not solely achieved by cell death in the developing chick.
During development of the projection from the retina to the brain in the chick, a transient ipsilateral retinotectal projection forms and disappears. This disappearance is coincident with a wave of ganglion cell death in the retina. The contribution of cell death to the disappearance of this projection, as opposed to another mechanism such as axon retraction, was examined. Retinal ganglion cells with a projection to the ipsilateral tectum were retrogradely labeled by injection of long-lasting fluorescent dyes into the tectum prior to the onset of ganglion cell death. Large injections of fast blue labeled approximately 1800 ganglion cells in the ipsilateral retina before the period of cell death began. If the injected embryos were allowed to survive past the peak period of ganglion cell death, the average number of labeled ganglion cells in the ipsilateral retina was reduced by somewhat more than half. It is possible that the remaining labeled ganglion cells projected only to nontectal visual nuclei and were labeled by fast blue that had diffused out of the tectum. This was tested by repeating the study using very localized injections of 1,1'-dioctodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate or fluorescent microspheres into the tectum. These small injections confirmed that cells with transient projections to the ipsilateral tectum survived past the elimination of this projection. Thus, ipsilaterally projecting ganglion cells have, at most, a slightly greater propensity for death than the average ganglion cell, and elimination of the transient ipsilateral retinotectal projection in chick can be explained only, in part, by the mechanism of cell death.(ABSTRACT TRUNCATED AT 250 WORDS)


GABAergic inhibition of histaminergic neurons regulates active waking but not the sleep-wake switch or propofol-induced loss of consciousness.
The activity of histaminergic neurons in the tuberomammillary nucleus (TMN) of the hypothalamus correlates with an animal's behavioral state and maintains arousal. We examined how GABAergic inputs onto histaminergic neurons regulate this behavior. A prominent hypothesis, the "flip-flop" model, predicts that increased and sustained GABAergic drive onto these cells promotes sleep. Similarly, because of the histaminergic neurons' key hub-like place in the arousal circuitry, it has also been suggested that anesthetics such as propofol induce loss of consciousness by acting primarily at histaminergic neurons. We tested both these hypotheses in mice by genetically removing ionotropic GABA(A) or metabotropic GABA(B) receptors from histidine decarboxylase-expressing neurons. At the cellular level, histaminergic neurons deficient in synaptic GABA(A) receptors were significantly more excitable and were insensitive to the anesthetic propofol. At the behavioral level, EEG profiles were recorded in nontethered mice over 24 h. Surprisingly, GABAergic transmission onto histaminergic neurons had no effect in regulating the natural sleep-wake cycle and, in the case of GABA(A) receptors, for propofol-induced loss of righting reflex. The latter finding makes it unlikely that the histaminergic TMN has a central role in anesthesia. GABA(B) receptors on histaminergic neurons were dispensable for all behaviors examined. Synaptic inhibition of histaminergic cells by GABA(A) receptors, however, was essential for habituation to a novel environment.


Development of a PET radioligand for potassium channels to image CNS demyelination.
Central nervous system (CNS) demyelination represents the pathological hallmark of multiple sclerosis (MS) and contributes to other neurological conditions. Quantitative and specific imaging of demyelination would thus provide critical clinical insight. Here, we investigated the possibility of targeting axonal potassium channels to image demyelination by positron emission tomography (PET). These channels, which normally reside beneath the myelin sheath, become exposed upon demyelination and are the target of the MS drug, 4-aminopyridine (4-AP). We demonstrate using autoradiography that 4-AP has higher binding in non-myelinated and demyelinated versus well-myelinated CNS regions, and describe a fluorine-containing derivative, 3-F-4-AP, that has similar pharmacological properties and can be labeled with 18F for PET imaging. Additionally, we demonstrate that [18F]3-F-4-AP can be used to detect demyelination in rodents by PET. Further evaluation in Rhesus macaques shows higher binding in non-myelinated versus myelinated areas and excellent properties for brain imaging. Together, these data indicate that [18F]3-F-4-AP may be a valuable PET tracer for detecting CNS demyelination noninvasively.


Effect of intrauterine infection and perinatal risk factors on serum concentrations of insulin like growth factor (IGF-I) in full-term and preterm newborns.
IGF-I is believed to be a key factor in fetal growth dynamics It is widely known, that serious early-onset infection in the newborn is a risk factor for further developmental disturbances in a child. However, effect of congenital infection as well as an influence of infectious and non-infectious perinatal risk factors on circulating IGF-I concentrations in newborns has not been examined, yet. Thus, the aim of this study was: 1) evaluation of IGF-I venous blood serum concentration in full-term and premature infants considering their sex, occurrence of intrauterine infection and perinatal risk factors; 2) establishing the relationship between IGF-I serum concentrations and chosen anthropometric parameters values in infected and healthy newborns. The study involved 112 newborns appropriate for gestational age. Taking into consideration occurrence of early onset infection and gestational age we divided examined children into 4 groups: I group--infected, full-term newborns; II group--infected premature newborns; III group--healthy full-term newborns; IV group--healthy premature newborns. In all infants immediately after birth anthropometric measurements were performed (birth weight, body length, circumference of head and circumference of chest) and serum IGF-I concentration was determined. We demonstrated that full-term infants with intrauterine infection have statistically significantly higher concentration of IGF-I in blood serum than infected premature infants and healthy full-term infants. Analysis of correlation revealed a significant positive linear correlations between IGF-I serum concentration and gestational age and anthropometric parameters values. We conclude that intrauterine infection increases serum IGF-I concentration in full-term infants, but not in preterm infants, that may be a result of immaturity. We suggest serum IGF-I concentration may be considered an additional element of developmental and nutritional state assessment in infected newborn.


Trends in out-of-pocket payments for health care in Kyrgyzstan, 2001-2007.
Within the countries of the former Soviet Union, the Kyrgyz Republic has been a pioneer in reforming the system of health care finance. Since the introduction of its compulsory health insurance fund in 1997, the country has gradually moved from subsidizing the supply of services to subsidizing the purchase of services through the 'single payer' of the health insurance fund. In 2002 the government introduced a new co-payment for inpatients along with a basic benefit package. A key objective of the reforms has been to replace the burgeoning system of unofficial informal payments for health care with a transparent official co-payment, thereby reducing the financial burden of health care spending for the poor. This article investigates trends in out-of-pocket payments for health care using the results of a series of nationally representative household surveys conducted over the period 2001-2007, when the reforms were being rolled out. The analysis shows that there has been a significant improvement in financial access to health care amongst the population. The proportion paying state providers for consultations fell between 2004 and 2007. As a result of the introduction of co-payments for hospital care, fewer inpatients report making payments to medical personnel, but when they are made, payments are high, especially to surgeons and anaesthetists. However, although financial access for outpatient care has improved, the burden of health care payments amongst the poor remains significant.


Audiological outcome measures with the BONEBRIDGE transcutaneous bone conduction hearing implant: impact of noise, reverberation and signal processing features.
Objective: To assess the performance of an active transcutaneous implantable-bone conduction device (TI-BCD), and to evaluate the benefit of device digital signal processing (DSP) features in challenging listening environments.Design: Participants were tested at 1- and 3-month post-activation of the TI-BCD. At each session, aided and unaided phoneme perception was assessed using the Ling-6 test. Speech reception thresholds (SRTs) and quality ratings of speech and music samples were collected in noisy and reverberant environments, with and without the DSP features. Self-assessment of the device performance was obtained using the Abbreviated Profile of Hearing Aid Benefit (APHAB) questionnaire.Study sample: Six adults with conductive or mixed hearing loss.Results: Average SRTs were 2.9 and 12.3 dB in low and high reverberation environments, respectively, which improved to -1.7 and 8.7 dB, respectively with the DSP features. In addition, speech quality ratings improved by 23 points with the DSP features when averaged across all environmental conditions. Improvement scores on APHAB scales revealed a statistically significant aided benefit.Conclusions: Noise and reverberation significantly impacted speech recognition performance and perceived sound quality. DSP features (directional microphone processing and adaptive noise reduction) significantly enhanced subjects' performance in these challenging listening environments.


Molecular analysis of fungal communities and laccase genes in decomposing litter reveals differences among forest types but no impact of nitrogen deposition.
The fungal community of the forest floor was examined as the cause of previously reported increases in soil organic matter due to experimental N deposition in ecosystems producing predominantly high-lignin litter, and the opposite response in ecosystems producing low-lignin litter. The mechanism proposed to explain this phenomenon was that white-rot basidiomycetes are more important in the degradation of high-lignin litter than of low-lignin litter, and that their activity is suppressed by N deposition. We found that forest floor mass in the low-lignin sugar-maple dominated system decreased in October due to experimental N deposition, whereas forest floor mass of high-lignin oak-dominated ecosystems was unaffected by N deposition. Increased relative abundance of basidiomycetes in high-lignin forest floor was confirmed by denaturing gradient gel electrophoresis (DGGE) and sequencing. Abundance of basidiomycete laccase genes, encoding an enzyme used by white-rot basidiomycetes in the degradation of lignin, was 5-10 times greater in high-lignin forest floor than in low-lignin forest floor. While the differences between the fungal communities in different ecosystems were consistent with the proposed mechanism, no significant effects of N deposition were detected on DGGE profiles, laccase gene abundance, laccase length heterogeneity profiles, or phenol oxidase activity. Our observations indicate that the previously detected accumulation of soil organic matter in the high-lignin system may be driven by effects of N deposition on organisms in the mineral soil, rather than on organisms residing in the forest floor. However, studies of in situ gene expression and temporal and spatial variability within forest floor communities will be necessary to further relate the ecosystem dynamics of organic carbon to microbial communities and atmospheric N deposition.


Understanding the dynamics of superparamagnetic particles under the influence of high field gradient arrays.
The aim of this study was to characterize the behaviour of superparamagnetic particles in magnetic drug targeting (MDT) schemes. A 3-dimensional mathematical model was developed, based on the analytical derivation of the trajectory of a magnetized particle suspended inside a fluid channel carrying laminar flow and in the vicinity of an external source of magnetic force. Semi-analytical expressions to quantify the proportion of captured particles, and their relative accumulation (concentration) as a function of distance along the wall of the channel were also derived. These were expressed in terms of a non-dimensional ratio of the relevant physical and physiological parameters corresponding to a given MDT protocol. The ability of the analytical model to assess magnetic targeting schemes was tested against numerical simulations of particle trajectories. The semi-analytical expressions were found to provide good first-order approximations for the performance of MDT systems in which the magnetic force is relatively constant over a large spatial range. The numerical model was then used to test the suitability of a range of different designs of permanent magnet assemblies for MDT. The results indicated that magnetic arrays that emit a strong magnetic force that varies rapidly over a confined spatial range are the most suitable for concentrating magnetic particles in a localized region. By comparison, commonly used magnet geometries such as button magnets and linear Halbach arrays result in distributions of accumulated particles that are less efficient for delivery. The trajectories predicted by the numerical model were verified experimentally by acoustically focusing magnetic microbeads flowing in a glass capillary channel, and optically tracking their path past a high field gradient Halbach array.


B cell repertoire diversity in athymic mice.
The extent of B cell repertoire diversity among nu/nu BALB/c mice has been assessed and compared with that of normal BALB/c mice. This was accomplished through the characterization of monoclonal, influenza hemagglutinin-specific antibodies by reactivity pattern analysis. The results indicate that the repertoire of athymic mice is equivalent in diversity to that of normal mice. Moreover, because these responses were obtained in recipients that were histocompatible but distinct at immunoglobulin allotype loci, these findings indicate that a very diverse array of B cell clonotypes may be stimulated in the absence of allotype-identical T cells.


Echo-planar perfusion MR of moyamoya disease.
Our goal was to assess the value of perfusion MR imaging by using a single-shot echo-planar technique to evaluate the hemodynamics of moyamoya disease. We performed echo-planar perfusion studies in 19 patients with a 1.5-T unit, using a free-induction-decay echo-planar sequence for 14 examinations and a turbo-gradient-spin-echo echo-planar sequence for five examinations. After a bolus injection of contrast material, 30 consecutive scans were done in 10 sections every 2 seconds. The data were analyzed to yield time-intensity curves for a region of interest set in the territory of the bilateral middle and/or anterior cerebral arteries in all examinations and to produce semiquantitative flow maps of each section, representing the signal decrease due to passage of contrast material in 17 examinations. The semiquantitative flow maps were compared with single-photon emission CT (SPECT) findings in 11 cases. We detected differences between the cerebral hemispheres and/or focal perfusion abnormalities by the time-intensity curves and semiquantitative flow maps in 15 of the 19 examinations and in 11 of the 17 examinations, respectively. Results of one or both these examinations corresponded with the SPECT findings in nine of the 11 examinations. Our results indicate that single-shot echo-planar perfusion MR imaging can sensitively depict hemodynamic abnormalities in moyamoya disease.


D(1) dopamine receptor stimulation increases GluR1 phosphorylation in postnatal nucleus accumbens cultures.
Postsynaptic interactions between dopamine and glutamate receptors in the nucleus accumbens are critical for acute responses to drugs of abuse and for neuroadaptations resulting from their chronic administration. We tested the hypothesis that D(1) dopamine receptor stimulation increases phosphorylation of the AMPA receptor subunit GluR1 at the protein kinase A phosphorylation site (Ser845). Nucleus accumbens cell cultures were prepared from postnatal day 1 rats. After 14 days in culture, GluR1 phosphorylation was measured by western blotting using phosphorylation site-specific antibodies. The D(1) receptor agonist SKF 81297 increased Ser845 phosphorylation in a concentration- dependent manner, with marked increases occurring within 5 min. This was prevented by the D(1) receptor antagonist SCH 23390 and the protein kinase A inhibitor H89, and reproduced by forskolin. The D(2) receptor agonist quinpirole attenuated the response to D(1) receptor stimulation. Neither D(1) nor D(2) receptor agonists altered GluR1 phosphorylation at Ser831, the site phosphorylated by protein kinase C and calcium/calmodulin-dependent protein kinase II. In other systems, phosphorylation of GluR1 at Ser845 is associated with enhancement of AMPA receptor currents. Thus, the present results suggest that AMPA receptor transmission in the nucleus accumbens may be augmented by concurrent D(1) receptor stimulation.


Association of Obesity-Related Hemodilution of Prostate-Specific Antigen, Dihydrotestosterone, and Testosterone.
Prostate-specific antigen (PSA) hemodilution is the leading theory for lower PSA values in obese men. However, testosterone and dihydrotestosterone (DHT), which are necessary for PSA production, are reduced in obese men. We assessed the relationship of body mass index (BMI) and PSA, taking into consideration the effect of testosterone and DHT. Among 8,122 participants in Reduction by Dutasteride of Prostate Cancer Events (REDUCE), complete data were available for 7,275. BMI was categorized as normal (<25 kg/m2 ), overweight (25-29.9 kg/m2 ), obese (30-34.9 kg/m2 ), or moderate + severely obese (≥35 kg/m2 ). Associations between BMI, testosterone, and DHT and the outcome variable of PSA were examined using linear regression. There were 1,964 (27.0%) normal weight, 3,826 (52.6%) overweight, 1,200 (16.5%) obese, and 285 (3.9%) moderately + severely obese patients. With increasing BMI, there was a progressive decrease in PSA (P = 0.02), increase in prostate volume (P < 0.001), and decrease in both testosterone (P < 0.001) and DHT (P < 0.001). Using linear regression, increasing BMI was associated with decreasing serum PSA values. Furthermore, BMI remained inversely associated with PSA after individually adjusting for testosterone and DHT, as well as when adjusting for testosterone and DHT in the same model. Decreased androgen levels accounted for only 19% of the lower PSA in men with higher BMI. Only a fraction of lower PSA in obese men could be attributed to testosterone and DHT levels. The remaining factors explaining lower PSA are unaccounted for, presumably secondary to hemodilution associated with increased plasma volume in obese men. Prostate 77:466-470, 2017. © 2016 Wiley Periodicals, Inc.