Proactively establishing a biologic specimens repository for large clinical trials: an idea whose time has come.
Large randomized clinical trials are becoming more costly, and resources to support them increasingly scarce. Biologic materials, such as blood, DNA, body fluids, or tissue samples collected and stored as a component of these studies represent an invaluable resource, to answer immediate secondary hypotheses, but also as archived material, linked to the study data, for the use of investigators long into the future. The regulatory climate surrounding the storage and future unconstrained utilization of biologic materials is evolving quickly. It is no longer acceptable simply to store samples and use them in an unbridled and unregulated fashion. Thus, to fully utilize the tremendous potential of biologic samples generated from large clinical trials and their related databases, investigators should consider proactively creating a biologic specimen repository, or biorepository. A repository likely assures appropriate subject consent, sample provenance, secure storage, and codified procedures for sample and data retrieval and sharing that protect the subject's confidentiality, the investigator's need for accurate data, and the limited resource. Importantly, the biorepository specimens/samples are typically collected in addition to local and core specimens obtained for the parent study that provide baseline assessments for safety and efficacy outcomes.


Dorsolateral prefrontal cortex contributes to the impaired behavioral adaptation in alcohol dependence.
Substance-dependent individuals often lack the ability to adjust decisions flexibly in response to the changes in reward contingencies. Prediction errors (PEs) are thought to mediate flexible decision-making by updating the reward values associated with available actions. In this study, we explored whether the neurobiological correlates of PEs are altered in alcohol dependence. Behavioral, and functional magnetic resonance imaging (fMRI) data were simultaneously acquired from 34 abstinent alcohol-dependent patients (ADP) and 26 healthy controls (HC) during a probabilistic reward-guided decision-making task with dynamically changing reinforcement contingencies. A hierarchical Bayesian inference method was used to fit and compare learning models with different assumptions about the amount of task-related information subjects may have inferred during the experiment. Here, we observed that the best-fitting model was a modified Rescorla-Wagner type model, the "double-update" model, which assumes that subjects infer the knowledge that reward contingencies are anti-correlated, and integrate both actual and hypothetical outcomes into their decisions. Moreover, comparison of the best-fitting model's parameters showed that ADP were less sensitive to punishments compared to HC. Hence, decisions of ADP after punishments were loosely coupled with the expected reward values assigned to them. A correlation analysis between the model-generated PEs and the fMRI data revealed a reduced association between these PEs and the BOLD activity in the dorsolateral prefrontal cortex (DLPFC) of ADP. A hemispheric asymmetry was observed in the DLPFC when positive and negative PE signals were analyzed separately. The right DLPFC activity in ADP showed a reduced correlation with positive PEs. On the other hand, ADP, particularly the patients with high dependence severity, recruited the left DLPFC to a lesser extent than HC for processing negative PE signals. These results suggest that the DLPFC, which has been linked to adaptive control of action selection, may play an important role in cognitive inflexibility observed in alcohol dependence when reinforcement contingencies change. Particularly, the left DLPFC may contribute to this impaired behavioral adaptation, possibly by impeding the extinction of the actions that no longer lead to a reward.


Systematic Review and Meta-analysis of Herpes Zoster Vaccine in Patients With CKD.
Chronic kidney disease (CKD) is a risk factor for herpes zoster (HZ) infection. Few studies have examined HZ vaccine (HZV) in this population. We conducted a systematic review and meta-analysis investigating the efficacy and safety of HZV in patients with renal disease (CKD, dialysis, and transplant). MEDLINE, Embase, and Cochrane Central Register of Controlled Trials (CENTRAL) databases (up to May 2020) were searched for randomized controlled trials and nonrandomized controlled studies evaluating HZV in patients with CKD for effectiveness and adverse event risks. Studies without a control group (placebo or no vaccine) were excluded. Extraction of prespecified data and risk of bias assessments using the Newcastle-Ottawa scale for cohort studies and the Cochrane Risk of Bias Tool for randomized controlled trials were done by 3 authors. Random-effects meta-analysis was used to generate pooled treatment effects and 95% confidence intervals. Included were 404,561 individuals from 8 studies (3 randomized controlled trials and 5 nonrandomized). All 8 studies examined HZ as an outcome, with 3 reporting adverse events. Risk of HZ was lower in patients who received HZV compared with controls (hazard ratio, 0.55; 95% confidence interval, 0.37-0.82; P < 0.01); however, heterogeneity was high (I 2 = 88%, P < 0.01). There was no significant difference in adverse events associated with HZV (hazard ratio, 1.03; 95% confidence interval, 0.54-1.28; P = 0.8). HZV compared with control significantly lowers the risk of HZ without an increase in adverse events in CKD patients. However, significant heterogeneity was present. HZV should be actively considered in CKD patients because the prevalence of HZ is higher in this population.


Inhibition of p25/Cdk5 Attenuates Tauopathy in Mouse and iPSC Models of Frontotemporal Dementia.
Increased p25, a proteolytic fragment of the regulatory subunit p35, is known to induce aberrant activity of cyclin-dependent kinase 5 (Cdk5), which is associated with neurodegenerative disorders, including Alzheimer's disease. Previously, we showed that replacing endogenous p35 with the noncleavable mutant p35 (Δp35) attenuated amyloidosis and improved cognitive function in a familial Alzheimer's disease mouse model. Here, to address the role of p25/Cdk5 in tauopathy, we generated double-transgenic mice by crossing mice overexpressing mutant human tau (P301S) with Δp35KI mice. We observed significant reduction of phosphorylated tau and its seeding activity in the brain of double transgenic mice compared with the P301S mice. Furthermore, synaptic loss and impaired LTP at hippocampal CA3 region of P301S mice were attenuated by blocking p25 generation. To further validate the role of p25/Cdk5 in tauopathy, we used frontotemporal dementia patient-derived induced pluripotent stem cells (iPSCs) carrying the Tau P301L mutation and generated P301L:Δp35KI isogenic iPSC lines using CRISPR/Cas9 genome editing. We created cerebral organoids from the isogenic iPSCs and found that blockade of p25 generation reduced levels of phosphorylated tau and increased expression of synaptophysin. Together, these data demonstrate a crucial role for p25/Cdk5 in mediating tau-associated pathology and suggest that inhibition of this kinase can remedy neurodegenerative processes in the presence of pathogenic tau mutation.SIGNIFICANCE STATEMENT Accumulation of p25 results in aberrant Cdk5 activation and induction of numerous pathological phenotypes, such as neuroinflammation, synaptic loss, Aβ accumulation, and tau hyperphosphorylation. However, it was not clear whether p25/Cdk5 activity is necessary for the progression of these pathological changes. We recently developed the Δp35KI transgenic mouse that is deficient in p25 generation and Cdk5 hyperactivation. In this study, we used this mouse model to elucidate the role of p25/Cdk5 in FTD mutant tau-mediated pathology. We also used a frontotemporal dementia patient-derived induced pluripotent stem cell carrying the Tau P301L mutation and generated isogenic lines in which p35 is replaced with noncleavable mutant Δp35. Our data suggest that p25/Cdk5 plays an important role in tauopathy in both mouse and human model systems.


Evidence for the indirect utilization of glucose for the synthesis of hepatic glycogen in man.
1. This study was designed to test the hypothesis that three-carbon intermediates can be used in the 'indirect' pathway of glycogen synthesis in human liver (i.e. a route additional to the use of glucose by the 'direct' pathway). 2. After an overnight fast, 13 patients were given an infusion of 20% (w/v) glucose before elective abdominal operation. All received a 2.5 g bolus of 2220 kBq of selectively 3H- and 14C-labelled glucose before removal of a 1 g biopsy of liver. 3H and 14C were determined in purified glycogen as well as in glucose and lactate from samples of peripheral blood. 3. The ratio and specific activities of 3H and 14C in glycogen were found to be significantly lower than those in administered glucose. By calculation, 7-74% of glycogen repletion occurred by indirect pathways and not all of this was from the glucose supplied. 4. This study suggests that the operation of a direct pathway in man is not exclusive and that significant repletion of hepatic glycogen occurs by an indirect route.


Education, survival, and avoidable deaths in Lithuanian cancer patients, 2001-2009.
Background Our aim in this study is to provide a systematic assessment of the site-specific cancer survival rates of patients with different educational levels, using population-based census-linked registry data covering the entire population of Lithuania. Material and methods The study is based on the linkage between all records of the 2001 population census and all records from Lithuanian Cancer Registry (cancer incidence) and Statistics Lithuania (deaths) for the period between 6 April 2001 and 31 December 2009. Results For the vast majority of cancer sites we found an inverse gradient in survival, with the worst survival indicators in the lowest educational group. We estimated that 18.6% of the deaths in Lithuanian cancer patients could have potentially been postponed, if all the patients had the same cancer mortality as the patients with the highest educational level. Conclusion Our findings offer a warning that although the survival rates of cancer patients are improving, this progress hides disparities between different groups of patients.


MicroRNA-132 dysregulation in Toxoplasma gondii infection has implications for dopamine signaling pathway.
Congenital toxoplasmosis and toxoplasmic encephalitis can be associated with severe neuropsychiatric symptoms. However, which host cell processes are regulated and how Toxoplasma gondii affects these changes remain unclear. MicroRNAs (miRNAs) are small noncoding RNA sequences critical to neurodevelopment and adult neuronal processes by coordinating the activity of multiple genes within biological networks. We examined the expression of over 1000 miRNAs in human neuroepithelioma cells in response to infection with Toxoplasma. MiR-132, a cyclic AMP-responsive element binding (CREB)-regulated miRNA, was the only miRNA that was substantially upregulated by all three prototype Toxoplasma strains. The increased expression of miR-132 was also documented in mice following infection with Toxoplasma. To identify cellular pathways regulated by miR-132, we performed target prediction followed by pathway enrichment analysis in the transcriptome of Toxoplasma-infected mice. This led us to identify 20 genes and dopamine receptor signaling was their strongest associated pathway. We then examined myriad aspects of the dopamine pathway in the striatum of Toxoplasma-infected mice 5days after infection. Here we report decreased expression of D1-like dopamine receptors (DRD1, DRD5), metabolizing enzyme (MAOA) and intracellular proteins associated with the transduction of dopamine-mediated signaling (DARPP-32 phosphorylation at Thr34 and Ser97). Increased concentrations of dopamine and its metabolites, serotonin (5-HT) and 5-hydroxyindoleacetic acid were documented by HPLC analysis; however, the metabolism of dopamine was decreased and 5-HT metabolism was unchanged. Our data show that miR-132 is upregulated following infection with Toxoplasma and is associated with changes in dopamine receptor signaling. Our findings provide a possible mechanism for how the parasite contributes to the neuropathology of infection.


Mechanisms of iPS cell generation and beyond.
The generation of induced pluripotent stem cells (iPSCs) achieved by overexpression of Oct4, Sox2, Klf4 and c-Myc, transformed our classical views of the cellular epigenetic landscape and delivered a new concept for cell and tissue engineering. In addition to iPSCs, several other cell types have also been generated by master transcription factor (TF)-mediated transdifferentiation. However, the critical molecular mechanisms amongst diverse cellular identity changes are not well understood. Through the investigation of reprogramming mechanisms, we recently revealed that over-expression of constitutive active Smad3 boosted not only iPSC generation, but also 3 other master TF-mediated conversions, from B cells to macrophages, myoblasts to adipocytes, and human fibroblasts to neurons. This demonstrated that there were common mechanisms underlying different master TF-mediated cell conversions. To illuminate such mechanisms further, we have recently performed CRISPR/Cas9-mediated genome-wide knockout screening during reprogramming with a lentiviral gRNA library containing 90,000 gRNAs. This screening provided us with ~15 novel reprogramming roadblock genes as well as ~20 candidate genes essential for the reprogramming process but not for ES cell self-renewal. This data set will be a valuable resource to further understand how overexpression of master TFs alters cellular identity, and to achieve more faithful, efficient cell conversions for regenerative medicine.(Presented at the 1934th Meeting, March 17, 2017).


Trends and geographic differentials in mortality under age 60 in India.
The study examines overall and region-specific mortality changes and regional mortality variation in India since the 1970s, using data from the Sample Registration System (SRS). An evaluation of the quality of SRS data confirms their reliability for children and adults under age 60. The results suggest the convergence of mortality across the regions of India with important inter-state differences in the pace of health improvements over time. After spectacular progress during the 1970s and the 1980s, many Indian states have witnessed slower mortality improvements in both young and adult age groups. India faces difficulties in making further reductions in infant mortality and in the burden of chronic and man-made diseases at adult ages.


Determinants of survival without antiretroviral therapy after infancy in HIV-1-infected Zambian children in the CHAP Trial.
There are few data on predictors of HIV progression in untreated children in resource-limited settings. Children with HIV Antibiotic Prophylaxis (CHAP) was a randomized trial comparing cotrimoxazole prophylaxis with placebo in HIV-infected Zambian children. The prognostic value of baseline characteristics was investigated using Cox models. Five hundred fourteen children aged 1 to 14 (median 5.5) years contributed 607 years follow-up (maximum 2.6 years). Half were boys, and in 67%, the mother was the primary carer; at baseline, median CD4 percentage was 11% and weight was less than third percentile in 67%. One hundred sixty-five children died (27.2 per 100 years at risk; 95% confidence interval 23.3-31.6). Low weight-for-age, CD4 percentage, hemoglobin, mother as primary carer, current malnutrition, and previous hospital admissions for respiratory tract infections or recurrent severe bacterial infections were independent predictors of poorer survival, whereas oral candidiasis predicted poorer survival only when baseline CD4 percentage was not considered. Mortality rates per 100 child years of 44.5 (37.2-53.2), 14.7 (10.9-19.8), and 2.3 (0.3-16.7) were associated with new World Health Organization stages 4, 3, and 1/2, respectively, applied retrospectively; very low weight-for-age was the only staging feature for 42% of stage 4 children. Malnutrition and hospitalizations for respiratory/bacterial infections predict mortality independent of immunosuppression, suggesting that they capture HIV- and non-HIV-related mortality, whereas oral candidiasis is a proxy for immunosuppression.


Updated response assessment criteria for high-grade gliomas: response assessment in neuro-oncology working group.
Currently, the most widely used criteria for assessing response to therapy in high-grade gliomas are based on two-dimensional tumor measurements on computed tomography (CT) or magnetic resonance imaging (MRI), in conjunction with clinical assessment and corticosteroid dose (the Macdonald Criteria). It is increasingly apparent that there are significant limitations to these criteria, which only address the contrast-enhancing component of the tumor. For example, chemoradiotherapy for newly diagnosed glioblastomas results in transient increase in tumor enhancement (pseudoprogression) in 20% to 30% of patients, which is difficult to differentiate from true tumor progression. Antiangiogenic agents produce high radiographic response rates, as defined by a rapid decrease in contrast enhancement on CT/MRI that occurs within days of initiation of treatment and that is partly a result of reduced vascular permeability to contrast agents rather than a true antitumor effect. In addition, a subset of patients treated with antiangiogenic agents develop tumor recurrence characterized by an increase in the nonenhancing component depicted on T2-weighted/fluid-attenuated inversion recovery sequences. The recognition that contrast enhancement is nonspecific and may not always be a true surrogate of tumor response and the need to account for the nonenhancing component of the tumor mandate that new criteria be developed and validated to permit accurate assessment of the efficacy of novel therapies. The Response Assessment in Neuro-Oncology Working Group is an international effort to develop new standardized response criteria for clinical trials in brain tumors. In this proposal, we present the recommendations for updated response criteria for high-grade gliomas.


The effects of oligomerization on Saccharomyces cerevisiae Mcm4/6/7 function.
Minichromosome maintenance proteins (Mcm) 2, 3, 4, 5, 6 and 7 are related by sequence and form a variety of complexes that unwind DNA, including Mcm4/6/7. A Mcm4/6/7 trimer forms one half of the Mcm2-7 hexameric ring and can be thought of as the catalytic core of Mcm2-7, the replicative helicase in eukaryotic cells. Oligomeric analysis of Mcm4/6/7 suggests that it forms a hexamer containing two Mcm4/6/7 trimers, however, under certain conditions trimeric Mcm4/6/7 has also been observed. The functional significance of the different Mcm4/6/7 oligomeric states has not been assessed. The results of such an assessment would have implications for studies of both Mcm4/6/7 and Mcm2-7. Here, we show that Saccharomyces cerevisiae Mcm4/6/7 reconstituted from individual subunits exists in an equilibrium of oligomeric forms in which smaller oligomers predominate in the absence of ATP. In addition, we found that ATP, which is required for Mcm4/6/7 activity, shifts the equilibrium towards larger oligomers, likely hexamers of Mcm4/6/7. ATPγS and to a lesser extent ADP also shift the equilibrium towards hexamers. Study of Mcm4/6/7 complexes containing mutations that interfere with the formation of inter-subunit ATP sites (arginine finger mutants) indicates that full activity of Mcm4/6/7 requires all of its ATP sites, which are formed in a hexamer and not a trimer. In keeping with this observation, Mcm4/6/7 binds DNA as a hexamer. The minimal functional unit of Mcm4/6/7 is a hexamer. One of the roles of ATP binding by Mcm4/6/7 may be to stabilize formation of hexamers.


Mechanism of stabilization of dicalcium silicate solid solution with aluminium.
Stoichiometric dicalcium silicate, Ca2SiO4, displays a well-known polymorphism with temperature. When this phase is doped by a range of elements, belite, one of the main phases of cements, is generated. Here, we thoroughly study the aluminum doping of dicalcium silicate. This type of study is important for cement characterization and also from a basic point of view. Ca2Si(1-2x)Al(2x)O(4-x)□(x) (x = 0, 0.010, 0.014, 0.03) has been prepared and studied by X-ray powder diffraction and the Rietveld method. The limiting composition has been established as Ca2Si0.972Al0.028O3.986□0.014. The (27)Al MAS NMR band located close to ~-70 ppm is ascribed to tetrahedral environments, in agreement with the proposed aliovalent Si/Al atomic substitution mechanism. Thermal analysis measurements under a wet atmosphere indirectly confirm the increase of oxygen vacancies as the amount of incorporated protons increases with the aluminium content. A thorough electrical characterization has been carried out including overall conductivity measurements under wet and dry atmospheres and conductivity as a function of the oxygen partial pressure. The samples show oxide anion conductivity with a small p-type electronic contribution under oxidizing conditions. These compounds display a very important proton contribution to the overall conductivities under humidified atmospheres.


Sharing the Now in the Social Present: Duration of Nonverbal Synchrony Is Linked With Personality.
The social present is a novel descriptor of dyadic nowness and social sharing, extending research on individual nowness (James's [1890] specious present) to the interpersonal and intersubjective domain. We wished to connect this descriptor to personality attributes. We define the social present by the duration of significant nonverbal synchrony, based on the phenomenon of movement synchrony that generally emerges in social interactions. It is thus an implicit and objective measure that can be implemented by automated video analyses. In this study, 168 healthy participants were invited to verbal conversations in same-sex dyads. We analyzed the associations of the social present with personality attributes and interaction types (competition, cooperation, fun task). The average duration of the social present was 6.0 seconds, highest in competitive interactions and in male-male dyads. People with higher Openness to Experience, higher avoidant attachment, and lower narcissistic interpersonal styles showed extended social present in their interactions. The concept of the social present extends personality attributes to the interpersonal domain and to intersubjectivity. The social present may be computed based on movement synchrony but also prosodic or physiological synchronies. We foresee implications for health-related interactions such as psychotherapy, where therapeutic presence is an essential property of alliance.


Pectoral fin locomotion in batoid fishes: undulation versus oscillation.
This study explores the dichotomy between undulatory (passing multiple waves down the fin or body) and oscillatory (flapping) locomotion by comparing the kinematics of pectoral fin locomotion in eight species of batoids (Dasyatis americana, D. sabina, D. say, D. violacea, Gymnura micrura, Raja eglanteria, Rhinobatos lentiginosus and Rhinoptera bonasus) that differ in their swimming behavior, phylogenetic position and lifestyle. The goals of this study are to describe and compare the pectoral fin locomotor behavior of the eight batoid species, to clarify how fin movements change with swimming speed for each species and to analyze critically the undulation/oscillation continuum proposed by Breder using batoids as an example. Kinematic data were recorded for each species over a range of swimming velocities (1-3 disc lengths s(-1)). The eight species in this study vary greatly in their swimming modes. Rhinobatos lentiginosus uses a combination of axial-based and pectoral-fin-based undulation to move forward through the water, with primary thrust generated by the tail. The pectoral fins are activated in short undulatory bursts for increasing swimming speed and for maneuvering. Raja eglanteria uses a combination of pectoral and pelvic locomotion, although only pectoral locomotion is analyzed here. The other six species use pectoral locomotion exclusively to propel themselves through the water. Dasyatis sabina and D. say have the most undulatory fins with an average of 1.3 waves per fin length, whereas Rhinoptera bonasus has the most oscillatory fin behavior with 0.4 waves per fin length. The remaining species range between these two extremes in the degree of undulation present on their fins. There is an apparent trade-off between fin-beat frequency and amplitude. Rhinoptera bonasus has the lowest frequency and the highest fin amplitude, whereas Rhinobatos lentiginosus has the highest frequency and the lowest amplitude among the eight species examined. The kinematic variables that batoids modify to change swimming velocity vary among different species. Rhinobatos lentiginosus increases its tail-beat frequency to increase swimming speed. In contrast, the four Dasyatis species increase swimming speed by increasing frequency and wavespeed, although D. americana also changes wave number. Raja eglanteria modifies its swimming velocity by changing wavespeed and wave number. Rhinoptera bonasus increases wavespeed, Gymnura micrura decreases wave number, and both Rhinoptera bonasus and Gymnura micrura increase fin-tip velocity to increase swimming velocity. Batoid species fall onto a continuum between undulation and oscillation on the basis of the number of waves present on the fins.


Limnological study on a newly built drinking water reservoir near Tirana, Albania.
Bovilla Lake is a reservoir constructed 12 years ago for supplying the city of Tirana (Albania) mainly with drinking water. It has a surface area of 4.6 km(2), a maximum depth of originally 60 m and is monomictic with a stratification period from early spring to end of October. The lake is oligotrophic with low nutrient concentrations (e.g. SRP in spring about 8 μg L( -1)) and minor oxygen depletion in the hypolimnion during thermal stagnation. The lake is highly turbid due to severe particle import by several rivers during rain periods. This led to a massive deposition of sediments, lifting the maximum depth to 45 m in 2008. Furthermore, the photic zone reached hardly more than 10 m. Algal species diversity is high; however, diatoms from the genus Cyclotella dominate most of the year both in numbers and biomass. Our study describes for the first time the hydrography and limnology of the Bovilla Reservoir.


Partial characterization of the shift from IgG to IgA synthesis in the clonal differentiation of human leukemic bone marrow-derived lymphocytes.
An unusual B-cell proliferation was noted in an individual (Tun) which was characterized by the presence of two separate populations of chronic lymphocytic leukemia (CLL) cell staining on the surface and in the cytoplasm for either IgG(k) or IgA(k). Utilizing an idiotypic antiserum prepared from the associated serum monoclonal IgG(k) protein the idiotype was detected on the surface and in the cytoplasm of both the IgG- and IgA-bearing cell populations. These observations are consistent with a common clonal origin and a switch mechanism involving IgG and IgA synthesis. Sequential-labeling of Surface Ig and intracellular Ig with antisera conjugated to opposite fluorochromes documented the progressive maturation of the terminal differentiation of the IgA-bearing cell population at a level before morphologically distinct plasma cells. The distribution and pattern of surface and cytoplasmic IgG and IgA staining in individual cells suggest that the direction of switching is from IgG to IgA synthesis. The demonstration of shared idiotypic specificity between the IgG- and IgA-bearing populations is consistent with a transition in Ig heavy chain synthesis resulting from an alternation in the CH gene. It is concluded that certain CLL clones may manifest a switch from IgG to IgA synthesis at a level of B-cell differentiation which encompasses both the B lymphocyte and the Ig-synthesizing plasma cell.


Tumor necrosis factor induces enhanced responses to platelet-activating factor and differentiation in human monocytic Mono Mac 6 cells.
Human Mono Mac 6 cells exhibit characteristics of mature blood monocytes. Treatment of these cells with human recombinant human tumor necrosis factor (TNF) resulted in an increase in phagocytosis and phorbol myristate acetate-stimulated superoxide anion production at 12 h and growth retardation occurring at 24 h. Moreover, TNF induced a moderate increase of CD14 surface antigen expression, used as a phenotypic marker of monocyte/macrophage differentiation. Platelet-activating factor (PAF) stimulated a rapid rise in cytosolic free Ca++ ([Ca++]i) of 308 +/- 93 nM in TNF-treated cells compared to untreated cells (33 +/- 8 nM, n = 4). The effect of TNF was dose and time dependent, evident after 12 h and maximal at 48 h. The enhanced PAF-induced [Ca++]i rise was inhibited by the PAF receptor antagonist L-659,989 and EGTA, indicating receptor-dependent Ca++ influx. Furthermore, L-659,989 and PAF inhibited specific 3H-labeled PAF binding in TNF-treated, but not in untreated cells. Consistently, PAF stimulated arachidonic acid release only in TNF-treated cells. Preincubation of cells with anti-TNF monoclonal antibodies abolished TNF-induced effects, but failed to block lipopolysaccharide (LPS) effects. Distinct mechanisms of action by LPS were reflected by the different ability to induce surface antigen expression. In conclusion, the enhancement of PAF responses by TNF, associated with functional characteristics of differentiation in Mono Mac 6 cells, may represent a specific mechanism of cooperative interaction between PAF and TNF in inflammation, sepsis, immunoregulation and atherogenesis.


A survey of quality of life indicators in the Romanian Roma population following the 'Decade of Roma Inclusion'.
Background: This study explores how the Roma in Romania, the EU's most concentrated population, are faring in terms of a number of quality of life indicators, including poverty levels, healthcare, education, water, sanitation, and hygiene. It further explores the role of synthetic populations and modelling in identifying at-risk populations and delivering targeted aid. Methods: 135 surveys were conducted across five geographically diverse Romanian communities. Household participants were selected through a comprehensive random walk method. Analyses were conducted on all data using Pandas for Python. Combining land scan data, time-use survey analyses, interview data, and ArcGIS, the resulting synthetic population was analysed via classification and regression tree (CART) analysis to identify hot-spots of need, both ethnically and geographically. Results: These data indicate that the Roma in Romania face significant disparities in education, with Roma students less likely to progress beyond 8 th grade. In addition, the Roma population remains significantly disadvantaged with regard to safe and secure housing, poverty, and healthcare status, particularly in connection to diarrheal disease. In contrast, however, both Roma and non-Roma in rural areas face difficulties regarding full-time employment, sanitation, and water, sanitation, and hygiene infrastructure. In addition, the use of a synthetic population can generate information about 'hot spots' of need, based on geography, ethnicity, and type of aid required. Conclusions: These data demonstrate the challenges that remain to the Roma population in Romania, and also point to the myriad of ways in which all rural Romanians, regardless of ethnicity, are encountering hardship. This study highlights an approach that combines traditional survey data with more wide-reaching geographically based data and CART analysis to determine 'hot spot' areas of need in a given population. With the appropriate inputs, this tool can be extrapolated to any population in any country.


Individuals with low working memory spans show greater interference from irrelevant information because of poor source monitoring, not greater activation.
Although individuals with high and low working memory (WM) span appear to differ in the extent to which irrelevant information interferes with their performance on WM tasks, the locus of this interference is not clear. The present study investigated whether, when performing a WM task, high- and low-span individuals differ in the activation of formerly relevant, but now irrelevant items, and/or in their ability to correctly identify such irrelevant items. This was done in two experiments, both of which used modified complex WM span tasks. In Experiment 1, the span task included an embedded lexical decision task designed to obtain an implicit measure of the activation of both currently and formerly relevant items. In Experiment 2, the span task included an embedded recognition judgment task designed to obtain an explicit measure of both item and source recognition ability. The results of these experiments indicate that low-span individuals do not hold irrelevant information in a more active state in memory than high-span individuals, but rather that low-span individuals are significantly poorer at identifying such information as irrelevant at the time of retrieval. These results suggest that differences in the ability to monitor the source of information, rather than differences in the activation of irrelevant information, are the more important determinant of performance on WM tasks.


[Clinical research progress of heterotopic ossification of elbow after injury].
To review the basic research and clinical progress of elbow heterotopic ossification after injury. The recent literature concerning heterotopic ossification of the elbow was reviewed. Heterotopic ossification was caused by variety of stimuli and conditions. The current methods of prevention and treatment were to improve surgical techniques, to reduce trauma and bleeding, to rinse the area with bone fragments with plenty of salt water, and to use non-steroidal anti-inflammatory drugs. Once heterotopic ossification occurred, surgical treatment is unique treatment method, so emphasis is to prevent heterotopic ossification.


Renin dependency of the effect of chronically administered atrial natriuretic factor in two-kidney, one clip rats.
Conscious two-kidney, one clip rats with 150 mm Hg or higher systolic blood pressure were infused with saralasin for 60 minutes. Those with a blood pressure decline of 30 mm Hg or more were classified as saralasin-sensitive; those with a decrease of 10 mm Hg or less were considered saralasin-resistant. The animals were then housed in metabolic cages. Groups of sham-operated normotensive, saralasin-sensitive or saralasin-resistant two-kidney, one clip (2K1C) rats were infused with atrial natriuretic factor (Arg 101-Tyr 126), 100 ng/hr per rat, for 6 days. Corresponding control groups were sham-infused. Blood pressure was initially higher in the saralasin-sensitive groups (176 +/- 6 and 181 +/- 1 mm Hg, respectively) than in the saralasin-resistant groups (160 +/- 4 and 169 +/- 4 mm Hg, respectively). Atrial natriuretic factor infusion produced a gradual decline in blood pressure to 128 +/- 5 mm Hg, but only in saralasin-sensitive 2K1C animals. Urinary volume, initially higher in saralasin-sensitive hypertensive than in normotensive rats, was depressed during atrial natriuretic factor infusion. Urinary sodium excretion and water intake showed the same tendency, but the changes were not significant. No such modifications were observed in saralasin-resistant or sham-operated rats infused with atrial natriuretic factor. Body weight, which was higher in normotensive animals, was unchanged during atrial natriuretic factor infusion. Saralasin-sensitive, noninfused 2K1C rats were the only group with higher plasma renin activity than sham-operated, normotensive controls. Plasma aldosterone was higher in the former than in the other five groups.(ABSTRACT TRUNCATED AT 250 WORDS)


The effect of low oxygen tension on the in vitro-replicative life span of human diploid fibroblast cells and their transformed derivatives.
Human diploid fibroblasts (HDF) IMR90, starting at various population doubling levels (PDL), were serially cultured under four different oxygen conditions; the conventional atmospheric 20% O2 condition and three lower oxygen conditions (1, 6, and 12% O2). All cultures from different PDLs showed that a longer replicative life span was achieved under the lower oxygen conditions. When the starting culture PDL was between 33 and 62, the increased life span rate was constant (average 22%) at 1% O2, but the rate decreased when the starting PDL was higher. The growth advantage under the 1% O2 condition was also observed in the very late passages of cultures to some extent, but terminal cultures with senescent cells were not stimulated by the low oxygen condition. When cultures at an extended PDL under the 1% O2 condition were shifted back to the 20% O2 condition, the cells rapidly senesced. HDF from a subject with Werner syndrome, a premature aging disease, which are known to have reduced replicative potential in vitro, also showed 43% increase in life span under the lowest oxygen conditions. When SV40 large T-transformed IMR90 cells at preimmortal stages were tested, no significant growth differences were observed under different oxygen conditions, and all the cultures died out at a similar PDL. These results suggest: (1) The atmospheric 20% oxygen tension hastens HDF senescence. (2) Young cells are more resistant to oxygen tension than old cells. (3) SV40 large T transformation abolishes the oxygen effect on cellular aging.


Targeted disruption of the Cl-/HCO3- exchanger Ae2 results in osteopetrosis in mice.
Osteoclasts are multinucleated bone-resorbing cells responsible for constant remodeling of bone tissue and for maintaining calcium homeostasis. The osteoclast creates an enclosed space, a lacuna, between their ruffled border membrane and the mineralized bone. They extrude H(+) and Cl(-) into these lacunae by the combined action of vesicular H(+)-ATPases and ClC-7 exchangers to dissolve the hydroxyapatite of bone matrix. Along with intracellular production of H(+) and HCO(3)(-) by carbonic anhydrase II, the H(+)-ATPases and ClC-7 exchangers seems prerequisite for bone resorption, because genetic disruption of either of these proteins leads to osteopetrosis. We aimed to complete the molecular model for lacunar acidification, hypothesizing that a HCO(3)(-) extruding and Cl(-) loading anion exchange protein (Ae) would be necessary to sustain bone resorption. The Ae proteins can provide both intracellular pH neutrality and serve as cellular entry mechanism for Cl(-) during bone resorption. Immunohistochemistry revealed that Ae2 is exclusively expressed at the contra-lacunar plasma membrane domain of mouse osteoclast. Severe osteopetrosis was encountered in Ae2 knockout (Ae2-/-) mice where the skeletal development was impaired with a higher diffuse radio-density on x-ray examination and the bone marrow cavity was occupied by irregular bone speculae. Furthermore, osteoclasts in Ae2-/- mice were dramatically enlarged and fail to form the normal ruffled border facing the lacunae. Thus, Ae2 is likely to be an essential component of the bone resorption mechanism in osteoclasts.


The diagnostic value of intracoronary optical coherence tomography.
Optical coherence tomography (OCT) is a novel light-based imaging modality for application in the coronary circulation. Compared to conventional intravascular ultrasound, OCT has a ten-fold higher image resolution. This advantage has seen OCT successfully applied in the assessment of atherosclerotic plaque, stent apposition, and tissue coverage, heralding a new era in intravascular coronary imaging. The present article discusses the diagnostic value of OCT, both in cardiovascular research as well as in potential clinical application.The unparalleled high image resolution and strong contrast between the coronary lumen and the vessel wall structure enable fast and reliable image interpretation. OCT makes it possible to visualize the presence of atherosclerotic plaque in order to characterize the structure and extent of coronary plaque and to quantify lumen dimensions, as well as the extent of lumen narrowing, in unprecedented detail. Based on optical properties, OCT is able to distinguish different tissue types, such as fibrous, lipid-rich, necrotic, or calcified tissue. Furthermore, OCT is able to cover the visualization of a variety of features of atherosclerotic plaques that have been associated with rapid lesion progression and clinical events, such as thin cap fibroatheroma, fibrous cap thickness, dense macrophage infiltration, and thrombus formation. These unique features allow the use of OCT to assess patients with acute coronary syndrome and to study the dynamic nature of coronary atherosclerosis in vivo and over time. This permits new insights into plaque progression, regression, and rupture, as well as the study of effects of therapies aimed at modulating these developments.Today's OCT technology allows high detail resolution as well as fast and safe clinical image acquisition. These unique features have established OCT as the gold standard for the assessment of coronary stents. This technique makes it possible to study stent expansion, peri-procedural vessel trauma, and the interaction of the stent with the vessel wall down to the level of individual stent struts, both acutely as well as in the long term, where it is has proven extremely sensitive to the detection of even minor amounts of tissue coverage. These qualities render OCT indispensable to addressing vexing clinical questions such as the relationship of drug-eluting stent deployment, vascular healing, the true time course of endothelial stent coverage, and late stent thrombosis. This may also better guide the optimal duration of dual anti-platelet therapy that currently remains unclear and relatively empirical.In the future, OCT might emerge, parallel to its undisputed position in research, as the tool of choice in all clinical scenarios where angiography is limited by its nature as a two-dimensional luminogram.


Unusual case of pyogenic spondylodiscitis, vertebral osteomyelitis and bilateral psoas abscesses after acupuncture: diagnosis and treatment with interventional management.
We report, for the first time, a case of pyogenic spondylodiscitis combined with vertebral osteomyelitis and bilateral psoas abscesses after acupuncture. A 60-year-old man was diagnosed with rectal cancer, and radical rectectomy and permanent colostomy were carried out. However, 3 years after the surgery the patient complained of pain in the lower back, and the symptoms worsened after seven sessions of acupuncture. Technetium 99m-labelled methylene diphosphonate ((99m)Tc-MDP) bone scintigraphy (BS) revealed abnormal uptake of (99m)Tc-MDP in the L4 and L5 vertebrae. He was admitted to our hospital because of suspected bone metastases from rectal cancer. He was diagnosed with infection based on a history of acupuncture and the findings of enhanced MRI and CT. Percutaneous lumbar discectomy (PLD), external drainage and irrigation using antibiotics were carried out to treat the L4-5 disc. Pathological analyses and bacterial culture of the resected disc confirmed infection with group C streptococcus. Postoperative antibiotic treatment resulted in significant pain relief on the third day and gradual complete relief. Considerable improvement was seen on CT and MRI at follow-up. We consider it highly likely that this patient's infection was caused by acupuncture. In patients with malignancy, abnormal uptake of (99m)Tc-MDP in BS may signify bone metastasis but can also be observed in bone infections. PLD can be used to resect diseased discs to relieve pain quickly and to prevent herniation of lumbar discs. After PLD, external drainage can be employed for abscess drainage, decompression and perfusion of antibiotics. PLD may serve as an alternative to open surgery for pyogenic spondylodiscitis.


Assessment of flat panel LCD primary class display performance based on AAPM TG 18 acceptance protocol.
The image display is an important component of the Picture Archiving and Communication System (PACS) and of digital imaging in general. In this paper, we assess the display performance of 32 different flat panel LCD devices, in terms of their reflection, luminance response, luminance uniformity, resolution, noise, veiling glare and color uniformity included in the tentative guidelines of the AAPM TG18 document version 8.1. We also report on the angular dependencies of luminance and contrast, which constitute one of the miscellaneous tests. The tools used included a telescopic photometer, which was also used as a colorimeter, an illuminance meter, light sources for the reflection assessment, light-blocking devices, and digital TG18 test patterns. The luminance ratio (LR), maximum luminance difference (ALmax) and deviation of contrast response with respect to that of DICOM GSDF were 379.2+/-61.0, 1.6+/-1.1%, and 4.84+/-0.58%, respectively. The maximum luminance nonuniformity was 9.2+/-3.9% for the 10% luminance of the TG18-UNL10 test pattern. In the luminance-based resolution test, the percent luminance difference (deltaL) at the center was 0.78+/-0.42%. In all cases of noise testing, the rectangular target in each square in the three quadrants was visible, as were all 15 targets, except for the smallest one, in each corner pattern and the center pattern. The glare ratio (GR) was 2350+/-1460. The average color uniformity parameter, delta(u',v'), across the display area of each display device was 0.002+/-0.001. Nevertheless, not all of the color uniformity parameters of the display devices associated with a workstation met the acceptance criteria. For 7 selected flat panel displays, the mean specular and diffuse reflection coefficients were 0.0061+/-0.0010 and 0.0017+/-0.0005 cd/m2 per lux, respectively. All of the test results conformed to the criteria recommended by AAPM TG18, indicating that the displays were fully acceptable for diagnostic image interpretation. The maximum viewing angle conforming to the DICOM 3.14 standard luminance responses with a 10% tolerance was found to be approximately 50 degrees in both directions along the vertical axis, 10 degrees in the upper direction and 20 degrees in the lower direction along the horizontal axis, and 20 degrees in the upper direction and 10 degrees in the lower direction along the diagonal axis. Therefore, a radiologist should interpret a displayed image by considering the physical characteristics of the narrow viewing angle of the AMLCD displays. The acceptance testing protocol described herein demonstrates the successful clinical implementation of the guidelines for the viewing conditions of medical displays, and if implemented with a QC program, can be used to determine when LCD devices used for diagnostic interpretation need to be upgraded.


Mental health status and community adjustment after treatment in a residential treatment program for homeless veterans.
An uncontrolled outcome study was conducted to examine clinical improvement and the relationship of psychiatric and substance abuse problems, community adjustment, and housing status among homeless veterans who participated in a multisite residential treatment program. The study was performed at three U.S. Department of Veterans Affairs medical centers in Florida, Ohio, and California. Baseline, discharge, and 3-month postdischarge follow-up data were collected for 255 veterans admitted to the Domiciliary Care for Homeless Veterans Program. Multiple dimensions of outcome were examined, including psychiatric symptoms, alcohol abuse, drug abuse, social contacts, income, employment, and housing. Program participation was found to be associated with improvement in all areas of mental health and community adjustment. Improvement in psychiatric symptoms was associated with superior housing outcomes and improvement in community adjustment. When correlates of improvement in alcohol and drug abuse were examined, only one of eight possible relationships was found to be significant: improvement in alcohol problems was positively associated with improvement in employment. Homeless mentally ill veterans derive clear benefits from participation in a multidimensional residential treatment program. Improvement in mental health problems, however, is weakly linked to improvement in other areas, suggesting that treatment programs may have to attend separately to multiple domains of life adjustment.


K-252a: a specific inhibitor of the action of nerve growth factor on PC 12 cells.
K-252a, a kinase inhibitor isolated from the culture broth of Nocardiopsis sp., selectively inhibits the actions of nerve growth factor (NGF) on PC 12 cells. At a concentration of 200 nM, K-252a prevents neurite generation initiated by NGF, but not neurite generation produced by fibroblast growth factor or outgrowth produced by dibutyryl cAMP. K-252a also inhibits the induction of ornithine decarboxylase by NGF, but stimulates ornithine decarboxylase induction by epidermal growth factor. Stimulation of phosphatidylinositol breakdown by NGF was similarly inhibited by K-252a, while stimulation by epidermal growth factor was enhanced. The NGF-induced decrease in the phosphorylation of a soluble protein, Nsp 100, was prevented by K-252a. K-252a blocks the NGF-induced heterodown-regulation of the epidermal growth factor receptor, but not the epidermal growth factor-induced homodown-regulation of the epidermal growth factor receptor. K-252a, then, provides a new tool for the dissection and study of NGF-requiring processes.


Identification and Distribution of Fungi Associated with Fairy Rings on Golf Putting Greens.
Traditional methods for identification of fairy ring fungi rely on the morphology of mature basidiocarps, which are ephemeral and often do not reach maturity on golf greens due to management practices. From 2007 to 2009, basidiocarps and soil samples were collected from 15 hybrid bermudagrass and 30 bentgrass greens exhibiting fairy ring symptoms in California, Florida, Hawaii, Illinois, Oklahoma, North Caroline, South Carolina, and Wisconsin. Genomic DNA was extracted from 122 unknown samples. Extractions were made from mycelium isolated from puffball or mushroom tissue, from mycelium isolated from a soil block, or through direct DNA extraction from infested soil. DNA also was extracted from 16 reference isolates. The internal transcribed spacer (ITS) region of ribosomal DNA was amplified and sequenced using the basidiomycete-specific primer sets ITS1f/ITS4b and Basid0001/2R. Phylogenetic trees were constructed with the neighborjoining algorithm, with nodes evaluated by bootstrap analysis. Most samples grouped into one of three clades corresponding to species within the family Lycoperdaceae: Arachnion album, Bovista dermoxantha, and Vascellum curtisii. Although over 60 different basidiomycetes have been associated with fairy rings in turfgrasses, relatively few species were found on golf putting greens in this study. Presently, DNA sequencing may be the most efficient method for attempting speciation of fairy ring fungi from infested soil.


Mobile devices for community-based REDD+ monitoring: a case study for Central Vietnam.
Monitoring tropical deforestation and forest degradation is one of the central elements for the Reduced Emissions from Deforestation and Forest Degradation in developing countries (REDD+) scheme. Current arrangements for monitoring are based on remote sensing and field measurements. Since monitoring is the periodic process of assessing forest stands properties with respect to reference data, adopting the current REDD+ requirements for implementing monitoring at national levels is a challenging task. Recently, the advancement in Information and Communications Technologies (ICT) and mobile devices has enabled local communities to monitor their forest in a basic resource setting such as no or slow internet connection link, limited power supply, etc. Despite the potential, the use of mobile device system for community based monitoring (CBM) is still exceptional and faces implementation challenges. This paper presents an integrated data collection system based on mobile devices that streamlines the community-based forest monitoring data collection, transmission and visualization process. This paper also assesses the accuracy and reliability of CBM data and proposes a way to fit them into national REDD+ Monitoring, Reporting and Verification (MRV) scheme. The system performance is evaluated at Tra Bui commune, Quang Nam province, Central Vietnam, where forest carbon and change activities were tracked. The results show that the local community is able to provide data with accuracy comparable to expert measurements (index of agreement greater than 0.88), but against lower costs. Furthermore, the results confirm that communities are more effective to monitor small scale forest degradation due to subsistence fuel wood collection and selective logging, than high resolution remote sensing SPOT imagery.


Genetic predisposition to increased blood cholesterol and triglyceride lipid levels and risk of Alzheimer disease: a Mendelian randomization analysis.
Although altered lipid metabolism has been extensively implicated in the pathogenesis of Alzheimer disease (AD) through cell biological, epidemiological, and genetic studies, the molecular mechanisms linking cholesterol and AD pathology are still not well understood and contradictory results have been reported. We have used a Mendelian randomization approach to dissect the causal nature of the association between circulating lipid levels and late onset AD (LOAD) and test the hypothesis that genetically raised lipid levels increase the risk of LOAD. We included 3,914 patients with LOAD, 1,675 older individuals without LOAD, and 4,989 individuals from the general population from six genome wide studies drawn from a white population (total n=10,578). We constructed weighted genotype risk scores (GRSs) for four blood lipid phenotypes (high-density lipoprotein cholesterol [HDL-c], low-density lipoprotein cholesterol [LDL-c], triglycerides, and total cholesterol) using well-established SNPs in 157 loci for blood lipids reported by Willer and colleagues (2013). Both full GRSs using all SNPs associated with each trait at p<5×10-8 and trait specific scores using SNPs associated exclusively with each trait at p<5 × 10-8 were developed. We used logistic regression to investigate whether the GRSs were associated with LOAD in each study and results were combined together by meta-analysis. We found no association between any of the full GRSs and LOAD (meta-analysis results: odds ratio [OR]=1.005, 95% CI 0.82-1.24, p = 0.962 per 1 unit increase in HDL-c; OR=0.901, 95% CI 0.65-1.25, p=0.530 per 1 unit increase in LDL-c; OR=1.104, 95% CI 0.89-1.37, p=0.362 per 1 unit increase in triglycerides; and OR=0.954, 95% CI 0.76-1.21, p=0.688 per 1 unit increase in total cholesterol). Results for the trait specific scores were similar; however, the trait specific scores explained much smaller phenotypic variance. Genetic predisposition to increased blood cholesterol and triglyceride lipid levels is not associated with elevated LOAD risk. The observed epidemiological associations between abnormal lipid levels and LOAD risk could therefore be attributed to the result of biological pleiotropy or could be secondary to LOAD. Limitations of this study include the small proportion of lipid variance explained by the GRS, biases in case-control ascertainment, and the limitations implicit to Mendelian randomization studies. Future studies should focus on larger LOAD datasets with longitudinal sampled peripheral lipid measures and other markers of lipid metabolism, which have been shown to be altered in LOAD. Please see later in the article for the Editors' Summary.


Atypical cerebral lateralisation in adults with compensated developmental dyslexia demonstrated using functional transcranial Doppler ultrasound.
Functional transcranial Doppler ultrasound (fTCD) is a relatively new and non-invasive technique that assesses cerebral lateralisation through measurements of blood flow velocity in the middle cerebral arteries. In this study fTCD was used to compare functional asymmetry during a word generation task between a group of 30 dyslexic adults and a group of 30 non-dyslexic individuals. In light of previous evidence of atypical laterality in dyslexia, a reduced leftward asymmetry was predicted and confirmed. We know from previous research that most people with atypical language lateralisation have normal language and literacy skills: nevertheless, our results confirm that language laterality is reduced in those with dyslexia. Theoretical explanations for this apparent conundrum are discussed.


Functional mapping of the disparate activities of the yeast moonlighting protein Hal3.
The Saccharomyces cerevisiae Hal3 protein is a moonlighting protein, able to function both as an inhibitory subunit of the Ppz1 protein phosphatase and as a constituent protomer of an unprecedented heterotrimeric PPCDC (phosphopantothenoylcysteine decarboxylase), the third enzyme of the CoA biosynthetic pathway. In the present study we initiated the dissection of the structural elements required for both disparate cellular tasks by using a combination of biochemical and genetic approaches. We show that the conserved Hal3 core [PD (PPCDC domain)] is necessary for both functions, as determined by in vitro and in vivo assays. The Hal3 NtD (N-terminal domain) is not functional by itself, although in vitro experiments indicate that when this domain is combined with the core it has a relevant function in Hal3's heteromeric PPCDC activity. Both the NtD and the acidic CtD (C-terminal domain) also appear to be important for Hal3's Ppz1 regulatory function, although our results indicate that the CtD fulfils the key role in this regard. Finally, we show that the introduction of two key asparagine and cysteine residues, essential for monofunctional PPCDC activity but absent in Hal3, is not sufficient to convert it into such a homomeric PPCDC, and that additional modifications of Hal3's PD aimed at increasing its resemblance to known PPCDCs also fails to introduce this activity. This suggests that Hal3 has undergone significant evolutionary drift from ancestral PPCDC proteins. Taken together, our work highlights specific structural determinants that could be exploited for full understanding of Hal3's cellular functions.


Distinct functional domains of the Salmonella enterica WbaP transferase that is involved in the initiation reaction for synthesis of the O antigen subunit.
WbaP is a membrane enzyme that initiates O antigen synthesis in Salmonella enterica by catalysing the transfer of galactose 1-phosphate (Gal-1-P) onto undecaprenyl phosphate (Und-P). WbaP possesses at least three predicted structural domains: an N-terminal region containing four transmembrane helices, a large central periplasmic loop, and a C-terminal domain containing the last transmembrane helix and a large cytoplasmic tail. In this work, we investigated the contribution of each region to WbaP function by constructing a series of mutant WbaP proteins and using them to complement O antigen synthesis in DeltawbaP mutants of S. enterica serovars Typhi and Typhimurium. Truncated forms of WbaP lacking the periplasmic loop exhibited altered chain-length distributions in O antigen polymerization, suggesting that this central domain is involved in modulating the chain-length distribution of the O polysaccharide. The N-terminal and periplasmic domains were dispensable for complementation of O antigen synthesis in vivo, suggesting that the C-terminal domain carries the sugar-phosphate transferase activity. However, despite the fact that they complemented the synthesis of O antigen in the DeltawbaP mutant in vivo, membrane extracts containing WbaP derivatives without the N-terminal domain failed to transfer radioactive Gal from UDP-Gal into a lipid-rich fraction. These results suggest that the N-terminal region of WbaP, which contains four transmembrane domains, is essential for the insertion or stability of the protein in the bacterial membrane. We propose that the domain structure of WbaP enables this protein not only to function in the transfer of Gal-1-P to Und-P but also to establish critical interactions with additional proteins required for the correct assembly of O antigen in S. enterica.


Defining the boundaries of normal thrombin generation: investigations into hemostasis.
In terms of its soluble precursors, the coagulation proteome varies quantitatively among apparently healthy individuals. The significance of this variability remains obscure, in part because it is the backdrop against which the hemostatic consequences of more dramatic composition differences are studied. In this study we have defined the consequences of normal range variation of components of the coagulation proteome by using a mechanism-based computational approach that translates coagulation factor concentration data into a representation of an individual's thrombin generation potential. A novel graphical method is used to integrate standard measures that characterize thrombin generation in both empirical and computational models (e.g max rate, max level, total thrombin, time to 2 nM thrombin ("clot time")) to visualize how normal range variation in coagulation factors results in unique thrombin generation phenotypes. Unique ensembles of the 8 coagulation factors encompassing the limits of normal range variation were used as initial conditions for the computational modeling, each ensemble representing "an individual" in a theoretical healthy population. These "individuals" with unremarkable proteome composition was then compared to actual normal and "abnormal" individuals, i.e. factor ensembles measured in apparently healthy individuals, actual coagulopathic individuals or artificially constructed factor ensembles representing individuals with specific factor deficiencies. A sensitivity analysis was performed to rank either individual factors or all possible pairs of factors in terms of their contribution to the overall distribution of thrombin generation phenotypes. Key findings of these analyses include: normal range variation of coagulation factors yields thrombin generation phenotypes indistinguishable from individuals with some, but not all, coagulopathies examined; coordinate variation of certain pairs of factors within their normal ranges disproportionately results in extreme thrombin generation phenotypes, implying that measurement of a smaller set of factors may be sufficient to identify individuals with aberrant thrombin generation potential despite normal coagulation proteome composition.


Self-induced transverse mode selection in a photorefractive extended cavity laser diode.
Previous works on photorefractive self-organizing laser cavities were about lasers that oscillate, prior the self-organization process occurs, on a set of axial modes sharing the same transverse structure. In a well-designed broad-area laser diode extended cavity, we theoretically and experimentally demonstrate that the insertion of a photorefractive crystal can also affect the transverse modal structure to force the laser, initially oscillating on several transverse modes, to oscillate on a single transverse and axial mode. This spatial self-organization process leads to an enhancement of the single mode operating range of the laser.


Energy Drinks: An Assessment of Their Market Size, Consumer Demographics, Ingredient Profile, Functionality, and Regulations in the United States.
The consumption of energy drinks is rapidly increasing, as demonstrated by their large market growth. The targeted demographic group is teenagers, young adults, 18 to 34 y old; although expansion into nontraditional markets is also occurring. It is claimed that energy drinks can offer an increased energy boost related to their ingredient profile of caffeine, taurine, herbal extracts, and vitamins. Research suggests that energy drink formulations, in addition to increasing energy utilization, may also improve mood, enhance physical endurance, reduce mental fatigue, and increase reaction time. However, in most cases, the corresponding mechanisms of action are not clear. In addition, concerns have been raised over their safety and with a currently weak regulatory environment, efforts need to be made to ensure consumer safety. The objective of this article is to review the current U.S. energy drink market with emphasis on its market size, target demographic, active ingredients, potential benefits, safety, and regulations.


Atypical McMurry cross-coupling reactions leading to a new series of potent antiproliferative compounds bearing the key [ferrocenyl-ene-phenol] motif.
In the course of the preparation of a series of ferrocenyl derivatives of diethylstilbestrol (DES), in which one of the 4-hydroxyphenyl moieties was replaced by a ferrocenyl group, the McMurry reaction of chloropropionylferrocene with a number of mono-aryl ketones unexpectedly yielded the hydroxylated ferrocenyl DES derivatives, 5a-c, in poor yields (10%-16%). These compounds showed high activity on the hormone-independent breast cancer cell line MDA-MB-231 with IC50 values ranging from 0.14 to 0.36 µM. Surprisingly, non-hydroxylated ferrocenyl DES, 4, showed only an IC50 value of 1.14 µM, illustrating the importance of the hydroxyethyl function in this promising new series. For comparison, McMurry reactions of the shorter chain analogue chloroacetylferrocene were carried out to see the difference in behaviour with mono-aryl ketones versus a diaryl ketone. The effect of changing the length of the alkyl chain adjacent to the phenolic substituent of the hydroxylated ferrocenyl DES was studied, a mechanistic rationale to account for the unexpected products is proposed, and the antiproliferative activities of all of these compounds on MDA-MB-231 cells lines were measured and compared. X-ray crystal structures of cross-coupled products and of pinacol-pinacolone rearrangements are reported.


Differences in expression of the cancer stem cell marker aldehyde dehydrogenase 1 among estrogen receptor-positive/human epidermal growth factor receptor type 2-negative breast cancer cases with early, late, and no recurrence.
The significance of the expression of aldehyde dehydrogenase 1 (ALDH1), a cancer stem cell marker, for predicting the recurrence of estrogen receptor (ER)-positive/human epidermal growth factor receptor type 2 (HER2)-negative breast cancer is still poorly understood. The value of ALDH1 in predicting the time of recurrence remains unknown. In total, 184 patients with early distant recurrence, 134 patients with late distant recurrence, and 321 control patients without recurrence for more than 10 years after starting initial treatment for ER-positive/HER2-negative breast cancer, registered in 9 institutions, were analyzed. We assessed relationships between ALDH1 and other clinicopathological features, and ALDH1 expression was compared among the three groups. The relationship between ALDH1 expression and overall survival after recurrence was also evaluated in each group. The rates of ALDH1 expression positivity (more than 1 %) in the early, late, and no recurrence groups were 18.4 %, 13.4 %, and 8.4 %, respectively. ALDH1 expression correlated significantly with lymph node metastases (p = 0.048) and the Ki-67 labeling index (p < 0.001) in the early recurrence group. Multivariate analysis revealed ALDH1 expression to be significantly higher in the early recurrence group than in the no recurrence group (adjusted OR 2.140, 95 % CI 1.144-4.003, p = 0.016). Moreover, there was a significant difference in ALDH1 expression between the early and no recurrence groups receiving adjuvant endocrine therapy and chemotherapy (adjusted OR 4.625, 95 % CI 1.881-12.474, p < 0.001). However, there was no difference in ALDH1 expression between the late and no recurrence groups in univariate analysis (OR 1.507, 95 % CI 0.738-2.998, p = 0.253). In multivariate analysis, ALDH1 was not a factor independently predicting overall survival after the detection of recurrence (adjusted OR 1.451, 95 % CI 0.985-2.085, p = 0.059). Among patients with ER-positive/HER2-negative breast cancer, ALDH1 expression was more common in those with early recurrence, and this expression was found to be associated with a more aggressive breast cancer phenotype than that in the patients without recurrence. Further study is needed to clarify the prognostic significance of the heterogeneity of cancer stem cells and to confirm their role in resistance to chemotherapy.


Food from faeces: Evaluating the efficacy of scat DNA metabarcoding in dietary analyses.
Scat DNA metabarcoding is increasingly being used to track the feeding ecology of elusive wildlife species. This approach has greatly increased the resolution and detection success of prey items contained in scats when compared with other classical methods. However, there have been few studies that have systematically tested the applicability and reliability of this approach to study the diet of large felids species in the wild. Here we assessed the effectiveness of this approach in the cheetah Acinonyx jubatus. We tested how scat degradation, meal size, prey species consumed and feeding day (the day a particular prey was consumed) influenced prey DNA detection success in captive cheetahs. We demonstrated that it is possible to obtain diet information from 60-day old scats using genetic approaches, but the efficiency decreased over time. Probability of species-identification was highest for food items consumed one day prior to scat collection and the probability of being able to identify the species consumed increased with the proportion of the prey consumed. Detection success varied among prey species but not by individual cheetah. Identification of prey species using DNA detection methods from a single consumption event worked for samples collected between 8 and 72 hours post-feeding. Our approach confirms the utility of genetic approaches to identify prey species in scats and highlight the need to account for the systematic bias in results to control for possible scat degradation, feeding day, meal size and prey species consumed especially in the wild-collected scats.


Control of Paratrichodorus allius and Corky Ringspot Disease in Potato with Shank-injected Metam Sodium.
Corky ringspot disease (CRS) of potato produces necrotic areas in tubers that are considered quality defects that can lead to crop rejection. CRS is caused by tobacco rattle virus that is vectored by stubby-root nematodes (Paratrichodorus spp., Trichodorus spp.) at very low population densities, making disease management difficult and expensive. Fumigation with metam sodium (MS) is a common practice to control soil-borne fungi and increase potato yield. MS is generally applied in water via chemigation (water-run, WR) but is ineffective at controlling CRS when WR-applied, even at high rates. Therefore, WR MS is often used in combination with 1,3-dichloropropene (1,3-D), aldicarb or oxamyl to attain adequate CRS control. Between 1996 and 2000, fields with a history of CRS were treated with WR MS, shank-injected MS, and/or 1,3-D, and tubers were evaluated for symptoms of CRS. Shank injection of MS (SH MS) at depths of 41 cm, 15 and 30 cm, or 15, 30 and 45 cm controlled CRS over 3 years of testing. All rates of 280 liters/ha or greater were effective. Shank injection of metam potassium (MP) at rates of 448 liters/ha was also effective. 1,3-D controlled CRS alone or in combination with WR or SH MS. Proper shank application of MS or MP may adequately control CRS without the additional cost of other nematicides at low (<10 P. allius/250 g soil) to moderate (10 to 30 P. allius/250 g soil) populations of the nematode vector. Although SH MS was superior to WR MS, additional research is necessary to determine if this practice would be sufficient at higher CRS disease pressure or if addition of other nematicides would be necessary.


CD4/CD8 ratio and CD8 counts predict CD4 response in HIV-1-infected drug naive and in patients on cART.
Plasma HIV viral load is related to declining CD4 lymphocytes. The extent to which CD8 cells, in addition to RNA viral load, predict the depletion of CD4 cells is not well characterized so far. We examine if CD8 cell count is a prognostic factor for CD4 cell counts during an HIV infection.A longitudinal analysis is conducted using data from the Swiss HIV cohort study collected between January 2000 and October 2014. Linear mixed regression models were applied to observations from HIV-1-infected treatment naive patients (NAIVE) and cART-treated patients to predict the short-term evolution of CD4 cell counts. For each subgroup, it was quantified to which extent CD8 cell counts or CD4/CD8 ratios are prognostic factors for disease progression.In both subgroups, 2500 NAIVE and 8902 cART patients, past CD4 cells are positively (P < 0.0001) and past viral load is negatively (P < 0.0001) associated with the outcome. Including additionally past CD8 cell counts improves the fit significantly (P < 0.0001) and increases the marginal explained variation 31.7% to 40.7% for the NAIVE and from 44.1% to 50.7% for the cART group. The past CD4/CD8 ratio (instead of the past CD8 level) is positively associated with the outcome, increasing the explained variation further to 41.8% for NAIVE and 51.9% for cART.


Sensitivity of elongation factor Tu (EF-Tu) from different bacterial species to the antibiotics efrotomycin, pulvomycin and MDL 62879.
The sensitivity of elongation factor Tu (EF-Tu) from different species of bacteria to the EF-Tu-binding antibiotics efrotomycin, pulvomycin and MDL 62879 was tested by measuring the effect of these antibiotics on cell-free protein synthesis systems. EF-Tu from four different Gram-negative species was sensitive to all three antibiotics. Among Gram-positive bacteria, EF-Tu of Bacillus subtilis, Staphylococcus aureus and Enterococcus faecalis was resistant to efrotomycin and less sensitive to pulvomycin than EF-Tu of Gram-negative bacteria. EF-Tus from streptococci were significantly less sensitive than EF-Tus from Gram-negative bacteria to both efrotomycin and pulvomycin. All of the EF-Tus were sensitive to MDL 62879. The same sensitivity pattern emerged from GDP exchange assays, performed with partially purified EF-Tu from different bacterial species and pure Escherichia coli EF-Ts. These results suggest that the site of action of MDL 62879 is more conserved among bacterial species than those of efrotomycin and pulvomycin. Heterogeneity of EF-Tus from different bacterial species was also reflected in differences in their apparent molecular masses estimated by SDS-PAGE. EF-Tus from the Gram-positive species had higher molecular masses than those from all but one of the Gram-negative species.


Sialendoscopy with holmium:YAG laser treatment for multiple large sialolithiases of the Wharton duct: a case report and literature review.
Sialolithiasis is defined as calcified stone(s) in the salivary duct or glands. Submandibular gland sialolithiasis is the most common (80 to 90%), followed by parotid gland sialolithiasis (5 to 15%). The typical clinical presentation is salivary gland swelling after eating. As the swelling persists, symptoms owing to local inflammation, such as pain and trismus, emerge. In severe cases, cellulitis and even abscess formation occur and subsequently lead to salivary gland atrophy or fistula formation if the sialolithiasis remains untreated. The most common treatment is complete excision of the affected gland together with the stone(s). In some cases, intraoral sialolithotomy is performed when the stone is solitary and easily palpable through the oral cavity. Sialendoscopy is increasingly performed because of its minimal invasiveness. The major limitation of endoscopic laser lithotripsy of the salivary glands is the size of the stone. Often, for a stone larger than 4 mm, multiple fragmentations of the stone into small pieces is necessary before the pieces can be removed by wire basket or grasping forceps. Recently, the holmium:YAG laser has been reported as quite effective in removing larger salivary gland stones. However, sialoendoscopic laser lithotripsy is a very time-consuming procedure and in most cases, when there are multiple large stones in a single gland, entire gland excision is recommended. This report describes a male patient diagnosed with multiple large stones in his left submandibular gland who was successfully treated under sialendoscopy with holmium:YAG laser lithotripsy.


Alterations in genes of the EGFR signaling pathway and their relationship to EGFR tyrosine kinase inhibitor sensitivity in lung cancer cell lines.
Deregulation of EGFR signaling is common in non-small cell lung cancers (NSCLC) and this finding led to the development of tyrosine kinase inhibitors (TKIs) that are highly effective in a subset of NSCLC. Mutations of EGFR (mEGFR) and copy number gains (CNGs) of EGFR (gEGFR) and HER2 (gHER2) have been reported to predict for TKI response. Mutations in KRAS (mKRAS) are associated with primary resistance to TKIs. We investigated the relationship between mutations, CNGs and response to TKIs in a large panel of NSCLC cell lines. Genes studied were EGFR, HER2, HER3 HER4, KRAS, BRAF and PIK3CA. Mutations were detected by sequencing, while CNGs were determined by quantitative PCR (qPCR), fluorescence in situ hybridization (FISH) and array comparative genomic hybridization (aCGH). IC50 values for the TKIs gefitinib (Iressa) and erlotinib (Tarceva) were determined by MTS assay. For any of the seven genes tested, mutations (39/77, 50.6%), copy number gains (50/77, 64.9%) or either (65/77, 84.4%) were frequent in NSCLC lines. Mutations of EGFR (13%) and KRAS (24.7%) were frequent, while they were less frequent for the other genes. The three techniques for determining CNG were well correlated, and qPCR data were used for further analyses. CNGs were relatively frequent for EGFR and KRAS in adenocarcinomas. While mutations were largely mutually exclusive, CNGs were not. EGFR and KRAS mutant lines frequently demonstrated mutant allele specific imbalance i.e. the mutant form was usually in great excess compared to the wild type form. On a molar basis, sensitivity to gefitinib and erlotinib were highly correlated. Multivariate analyses led to the following results: 1. mEGFR and gEGFR and gHER2 were independent factors related to gefitinib sensitivity, in descending order of importance. 2. mKRAS was associated with increased in vitro resistance to gefitinib. Our in vitro studies confirm and extend clinical observations and demonstrate the relative importance of both EGFR mutations and CNGs and HER2 CNGs in the sensitivity to TKIs.


Ocular gene transfer with self-complementary AAV vectors.
Self-complementary AAV (scAAV) vectors have been developed to circumvent rate-limiting second-strand synthesis in single-stranded AAV vector genomes and to facilitate robust transgene expression at a minimal dose. In this study, the authors investigated the effects of intraocular injections of type 2 scAAV.GFP in mice. Dose-response experiments were performed to compare conventional single-strand AAV type 2 (ssAAV2) vectors with scAAV2 vectors encoding an identical expression cassette. Subretinal injection of 5 x 10(8) viral particles (vp) of scAAV.CMV-GFP resulted in green fluorescent protein (GFP) expression in almost all retinal pigment epithelial (RPE) cells within the area of the small detachment caused by the injection by 3 days and strong, diffuse expression by 7 days. Expression was strong in all retinal cell layers by days 14 and 28. In contrast, 3 days after subretinal injection of 5 x 10(8) vp of ssAAV.CMV-GFP, GFP expression was detectable in few RPE cells. Moreover, the ssAAV vector required 14 days for the attainment of expression levels comparable to those observed using scAAV at day 3. Expression in photoreceptors was not detectable until day 28. Dose-response experiments confirmed that onset of GFP expression was more rapid and robust after subretinal injection of scAAV.CMV-GFP than of ssAAV.CMV-GFP, resulting in pronounced expression in photoreceptors and other retinal neurons. Similar results were obtained for intravitreous injections. These data suggest that scAAV vectors may be advantageous for ocular gene therapy, particularly in retinal diseases that require rapid and robust transgene expression in photoreceptor cells.


Polarization properties of a triangular ring laser having a discharge tube with brewster angle windows.
The polarization properties of ring resonators for the case of a single discharge tube with Brewster angle windows in the optical circuit are investigated. The azimuths of polarization of the traveling waves and the related reflection losses due to the Brewster angle windows are calculated as a function of the orientation angle of the Brewster angle windows for different anisotropic reflectances of the reflecting mirrors of the resonators. The experiments are made on a triangular ring laser which is operated at the 6328 A He-Ne line. The measured polarization and intensity of the output light agree very well with the theoretical predictions.


Establishment and validation of a risk prediction model in patients with hepatocellular carcinoma treated with transarterial radioembolization.
Few studies have reported the treatment outcomes of transarterial radioembolization (TARE) using yttrium-90 (Y) for hepatocellular carcinoma (HCC). We established and validated a new risk prediction model for patients with HCC treated with TARE. Between 2010 and 2017, 113 and 35 patients with intrahepatic HCC treated with TARE were selected for the training and validation cohorts, respectively. The modified response evaluation criteria in solid tumors (mRECIST) were used for response evaluation. In the training cohort, the median age was 64.1 years (92 males and 21 females) and the mean survival after TARE was 50.3 months. The cumulative survival rates at six and 12 months were 92.0 and 84.0%, respectively. A new risk prediction model for patients with HCC treated with TARE (Y-scoring system) was established from the training cohort using five independent baseline variables [serum albumin < 3.5 g/dL, hazard ratio = 5.446; alpha-fetoprotein > 200 ng/mL (hazard ratio = 5.071); tumor number ≥ 3 (hazard ratio = 2.933); portal vein thrombosis (hazard ratio = 4.915); and hepatic vein invasion (hazard ratio = 8.500)] and two on-treatment variables [no des-gamma-carboxy prothrombin response (hazard ratio = 15.346) and progressive disease at three months (hazard ratio = 4.154)] for mortality (all P < 0.05). The predictive accuracy of the Y-scoring system was acceptable to predict six [area under the curve (AUC) = 0.845], nine (AUC = 0.868), and 12-month mortality (AUC = 0.886) (all P < 0.05). The predictive accuracy of the system was similarly maintained in the validation cohort (AUC 0.737-0.901 at 6-12 months). Our new risk prediction model can be used to stratify different prognoses in patients with HCC treated with TARE. Validation studies are required.


Quantitative assessment of distance to collection point and improved sorting information on source separation of household waste.
The present study measures the participation of households in a source separation scheme and, in particular, if the household's application of the scheme improved after two interventions: (a) shorter distance to the drop-off point and (b) easy access to correct sorting information. The effect of these interventions was quantified and, as far as possible, isolated from other factors that can influence the recycling behaviour. The study was based on households located in an urban residential area in Sweden, where waste composition studies were performed before and after the interventions by manual sorting (pick analysis). Statistical analyses of the results indicated a significant decrease (28%) of packaging and newsprint in the residual waste after establishing a property close collection system (intervention (a)), as well as significant decrease (70%) of the miss-sorted fraction in bags intended for food waste after new information stickers were introduced (intervention (b)). Providing a property close collection system to collect more waste fractions as well as finding new communication channels for information about sorting can be used as tools to increase the source separation ratio. This contribution also highlights the need to evaluate the effects of different types of information and communication concerning sorting instructions in a property close collection system.


Analysis of factors important for transurethral ultrasound prostate heating using MR temperature feedback.
The feasibility of using MR thermometry for temperature feedback to control a transurethral ultrasound heating applicator with planar transducers was investigated. The sensitivity of a temperature-based feedback algorithm to spatial (control point area, slice thickness, angular alignment) and non-spatial (imaging time, temperature uncertainty) parameters was evaluated through numerical simulations. The angular alignment of the control point with the ultrasound beam was an important parameter affecting the average spatial error in heat delivery. The other spatial parameters were less influential, thus providing an opportunity to reduce spatial resolution for increased SNR in the MR imaging. The update time was the most important non-spatial parameter determining the performance of the control algorithm. Combined non-spatial and spatial parameters achieved acceptable performance with a voxel size of 3 mm x 3 mm, a 10 mm slice thickness and a 5 s update time. Temperature uncertainty of up to 2 degrees C had little effect on the performance of the control algorithm but did reduce the average error slightly due to a systematic, noise-induced overestimation of the boundary temperature. These simulations imply that MR thermometry performed on clinical 1.5 T imaging systems is of sufficient quality for use as thermal feedback for conformal prostate thermal therapy with transurethral ultrasound heating applicators incorporating planar transducers.


Nuclear envelope breakdown is under nuclear not cytoplasmic control in sea urchin zygotes.
Nuclear envelope breakdown (NEB) and entry into mitosis are though to be driven by the activation of the p34cdc2-cyclin B kinase complex or mitosis promoting factor (MPF). Checkpoint control mechanisms that monitor essential preparatory events for mitosis, such as DNA replication, are thought to prevent entry into mitosis by downregulating MPF activation until these events are completed. Thus, we were surprised to find that when pronuclear fusion in sea urchin zygotes is blocked with Colcemid, the female pronucleus consistently breaks down before the male pronucleus. This is not due to regional differences in the time of MPF activation, because pronuclei touching each other break down asynchronously to the same extent. To test whether NEB is controlled at the nuclear or cytoplasmic level, we activated the checkpoint for the completion of DNA synthesis separately in female and male pronuclei by treating either eggs or sperm before fertilization with psoralen to covalently cross-link base-paired strands of DNA. When only the maternal DNA is cross-linked, the male pronucleus breaks down first. When the sperm DNA is cross-linked, male pronuclear breakdown is substantially delayed relative to female pronuclear breakdown and sometimes does not occur. Inactivation of the Colcemid after female NEB in such zygotes with touching pronuclei yields a functional spindle composed of maternal chromosomes and paternal centrosomes. The intact male pronucleus remains located at one aster throughout mitosis. In other experiments, when psoralen-treated sperm nuclei, over 90% of the zygote nuclei do not break down for at least 2 h after the controls even though H1 histone kinase activity gradually rises close to, or higher than, control mitotic levels. The same is true for normal zygotes treated with aphidicolin to block DNA synthesis. From these results, we conclude that NEB in sea urchin zygotes is controlled at the nuclear, not cytoplasmic, level, and that mitotic levels of cytoplasmic MPF activity are not sufficient to drive NEB for a nucleus that is under checkpoint control. Our results also demonstrate that the checkpoint for the completion of DNA synthesis inhibits NEB by acting primarily within the nucleus, not by downregulating the activity of cytoplasmic MPF.


Retrospective comparative evaluation of the lasting impact of a community-based primary health care programme on under-5 mortality in villages around Jamkhed, India.
To conduct the first rigorous evaluation of the long-term effect of the Comprehensive Rural Health Project on childhood mortality in rural Maharashtra. Background information and full birth histories were collected by conducting household surveys and interviewing women. Control villages resembling project villages in terms of population size were randomly selected from an area enclosed by two ellipses centred around, but not including, the project area. An equal number of villages and approximately equal numbers of households and women were randomly sampled from both areas. Cox models with robust standard errors were used to compare the hazard of death among children under 5 years of age in project and control villages. The hazard of death was reduced by 30% (95% confidence interval, CI: 6% to 48%) after the neonatal period in the project villages compared with control villages after adjustment for caste and religion of subjects and for availability of irrigation in the villages. During the neonatal period there was an increase of 3% in the hazard of death, but it was not statistically significant (95% CI: -18% to 29%). Our methods provide useful tools for evaluating long-running community-based primary health care programmes. Our findings add to the growing debate on the long-term sustainability of community-based interventions designed to reduce child mortality.


Geriatric Prescription in a Nigerian Tertiary Hospital.
To assess the medications prescribed for elderly inpatients on specific days during hospital admission with a view to detecting areas of irrational prescription. It was a prospective study of all patients aged 65 years and above admitted to the medical wards of a Nigerian tertiary hospital over a 12-month period. The World Health Organization/International Network of Rational Use of Drugs (WHO/INRUD) drug use indicators were used to assess drug prescriptions on various days of admission. A total of 1513 patient encounters involving 345 patients aged between 65 and 92 years were assessed on hospital days 1, 3, 5, 7, 14, and 28. The average number of medicines per encounter ranged from 6.1 ± 2.5 on hospital day 1 to 7.8 ± 2.4 on hospital day 28. This difference was statistically significant (F = 14.42; P < 0.05). The percentage of encounters with an antibiotic prescribed ranged from 50.4% on hospital day 1 to 62.9% on hospital day 28 while the percentage of encounters with an injection prescribed decreased from 72.8% on hospital day 1 to 50.0% on day 28. This study suggests some degree of irrational prescribing as evident by the high average number of medicine per encounter and the high percentages of encounters with an antibiotic or injection prescribed. However, there is a need to develop standard values for the WHO/INRUD indicators based on the recently published national treatment guidelines for common elderly diseases which will serve as yardsticks to assess elderly inpatients prescriptions using WHO/INRUD core indicators in future studies.


Global cerebral ischemia: effects of pentastarch after reperfusion.
There has been considerable interest in the use of synthetic hydroxyethyl starch macromolecules (e.g., pentastarch, a colloid used for intravascular fluid replacement) to reduce microvascular permeability and reperfusion injury after cerebral ischemia. A recent report found that hemodilution with pentastarch reduced brain injury and cerebral edema after temporary focal ischemia. We compared the effects of pentastarch versus 0.9% saline on brain edema after reperfusion in a model of temporary global ischemia in halothane-anesthetized rabbits. To ensure the validity of our model, we studied an additional group of animals in which we deliberately raised plasma osmolality with hypertonic saline (1.5%) in the expectation of finding a decreased brain water content at the conclusion of the experiment. Animals were hemodiluted to a hematocrit of 20% with normal saline (control group) (n = 9), pentastarch (n = 7), or hypertonic saline (n = 5). After hemodilution, the animals underwent a 25-min period of global cerebral ischemia, followed by 180 min of reperfusion. The animals were then killed and brain water content was assessed by microgravimetry and by the wet-dry weight method. As anticipated, colloid osmotic pressure was maintained in the pentastarch group, and plasma osmolality became significantly increased in the hypertonic saline group. There were no intergroup differences at any time for central venous pressure, mean arterial pressure, intracranial pressure, or PaCO2. Brain water content was significantly decreased in the hypertonic saline group. No difference in brain water content was detected between the control group and the pentastarch group.(ABSTRACT TRUNCATED AT 250 WORDS)


Pronounced peptide selectivity for melanoma through tryptophan end-tagging.
Effects of oligotryptophan end-tagging on the uptake of arginine-rich peptides into melanoma cells was investigated under various conditions and compared to that into non-malignant keratinocytes, fibroblasts, and erythrocytes, also monitoring resulting cell toxicity. In parallel, biophysical studies on peptide binding to, and destabilization of, model lipid membranes provided mechanistic insight into the origin of the selectivity between melanoma and non-malignant cells. Collectively, the results demonstrate that W-tagging represents a powerful way to increase selective peptide internalization in melanoma cells, resulting in toxicity against these, but not against the non-malignant cells. These effects were shown to be due to increased peptide adsorption to the outer membrane in melanoma cells, caused by the presence of anionic lipids such as phosphatidylserine and ganglioside GM1, and to peptide effects on mitochondria membranes and resulting apoptosis. In addition, the possibility of using W-tagged peptides for targeted uptake of nanoparticles/drug carriers in melanoma was demonstrated, as was the possibility to open up the outer membrane of melanoma cells in order to facilitate uptake of low Mw anticancer drugs, here demonstrated for doxorubicin.


Mouth Rinsing with Maltodextrin Solutions Fails to Improve Time Trial Endurance Cycling Performance in Recreational Athletes.
The carbohydrate (CHO) concentration of a mouth rinsing solution might influence the CHO sensing receptors in the mouth, with consequent activation of brain regions involved in reward, motivation and regulation of motor activity. The purpose of the present study was to examine the effects of maltodextrin mouth rinsing with different concentrations (3%, 6% and 12%) after an overnight fast on a 20 km cycling time trial performance. Nine recreationally active, healthy males (age: 24 ± 2 years; V ˙ O 2 m a x : 47 ± 5 mL·kg(-1)·min(-1)) participated in this study. A double-blind, placebo-controlled randomized study was conducted. Participants mouth-rinsed every 2.5 km for 5 s. Maltodextrin mouth rinse with concentrations of 3%, 6% or 12% did not change time to complete the time trial and power output compared to placebo (p > 0.05). Time trial completion times were 40.2 ± 4.0, 40.1 ± 3.9, 40.1 ± 4.4, and 39.3 ± 4.2 min and power output 205 ± 22, 206 ± 25, 210 ± 24, and 205 ± 23 W for placebo, 3%, 6%, and 12% maltodextrin conditions, respectively. Heart rate, lactate, glucose, and rating of perceived exertion did not differ between trials (p > 0.05). In conclusion, mouth rinsing with different maltodextrin concentrations after an overnight fast did not affect the physiological responses and performance during a 20 km cycling time trial in recreationally active males.


Speeding-up while growing-up: Synchronous functional development of motor and non-motor processes across childhood and adolescence.
Describing the maturation of information processing in children is fundamental for developmental science. Although non-linear changes in reaction times have been well-documented, direct measurement of the development of the different processing components is lacking. In this study, electromyography was used to quantify the maturation of premotor and motor processes on a sample of 114 children (6-14 years-old) and 15 adults. Using a model-based approach, we show that the development of these two components is well-described by an exponential decrease in duration, with the decay rate being equal for the two components. These findings provide the first unbiased evidence in favour of the common developmental rate of nonmotor and motor processes by directly confronting rates of development of different processing components within the same task. This common developmental rate contrasts with the differential physical maturation of region-specific cerebral gray and white matter. Tentative paths of interpretation are proposed in the discussion.