Chronotherapeutic treatments for depression in youth.
Chronotherapeutics such as wake therapy and bright light therapy are well-established methods in treating adults with depressive disorders and are additionally beneficent for sleep regulation. Few studies concerning chronotherapeutics in juvenile depression exist, though the established treatments are insufficient and sleep disorders often co-occur. In this study, we investigate the impact of two types of chronotherapeutics on depressive symptoms and sleep behavior in a juvenile setting. Juvenile inpatients (n = 62) with moderate to severe depressive symptoms took part in either a combined setting consisting of one night wake therapy followed by 2 weeks bright light therapy or in a setting of bright light therapy alone. Depressive symptoms, general psychopathology, clinical impression and sleep behavior were measured before (T1), directly after (T2) and 2 weeks after intervention (T3). Depressive symptoms decreased while sleep quality increased in both groups. The bright light therapy alone group showed further improvement at T3 in regards to depressive symptoms. Correlation analyses indicated significant negative correlations between sleep quality and awaking after restorative sleep with the depressive symptoms. However, only awaking after restorative sleep had a predictive impact on treatment outcome. The present study provides first evidence for a positive impact of chronotherapeutic interventions on treatment outcome in depressed juvenile inpatients. Bright light therapy seems to stabilize and further enhance reduction of depressive symptoms during follow-up, whereas one night wake therapy does not have an additional long-lasting impact on depressive symptoms and sleep parameters.


Occurrence of autoantibodies to annexin I, 14-3-3 theta and LAMR1 in prediagnostic lung cancer sera.
We have implemented a high throughput platform for quantitative analysis of serum autoantibodies, which we have applied to lung cancer for discovery of novel antigens and for validation in prediagnostic sera of autoantibodies to antigens previously defined based on analysis of sera collected at the time of diagnosis. Proteins from human lung adenocarcinoma cell line A549 lysates were subjected to extensive fractionation. The resulting 1,824 fractions were spotted in duplicate on nitrocellulose-coated slides. The microarrays produced were used in a blinded validation study to determine whether annexin I, PGP9.5, and 14-3-3 theta antigens previously found to be targets of autoantibodies in newly diagnosed patients with lung cancer are associated with autoantibodies in sera collected at the presymptomatic stage and to determine whether additional antigens may be identified in prediagnostic sera. Individual sera collected from 85 patients within 1 year before a diagnosis of lung cancer and 85 matched controls from the Carotene and Retinol Efficacy Trial (CARET) cohort were hybridized to individual microarrays. We present evidence for the occurrence in lung cancer sera of autoantibodies to annexin I, 14-3-3 theta, and a novel lung cancer antigen, LAMR1, which precede onset of symptoms and diagnosis. Our findings suggest potential utility of an approach to diagnosis of lung cancer before onset of symptoms that includes screening for autoantibodies to defined antigens.


Membrane potential dynamics of GABAergic neurons in the barrel cortex of behaving mice.
Computations in cortical circuits are mediated by synaptic interactions between excitatory and inhibitory neurons, and yet we know little about their activity in awake animals. Here, through single and dual whole-cell recordings combined with two-photon microscopy in the barrel cortex of behaving mice, we directly compare the synaptically driven membrane potential dynamics of inhibitory and excitatory layer 2/3 neurons. We find that inhibitory neurons depolarize synchronously with excitatory neurons, but they are much more active with differential contributions of two classes of inhibitory neurons during different brain states. Fast-spiking GABAergic neurons dominate during quiet wakefulness, but during active wakefulness Non-fast-spiking GABAergic neurons depolarize, firing action potentials at increased rates. Sparse uncorrelated action potential firing in excitatory neurons is driven by fast, large, and cell-specific depolarization. In contrast, inhibitory neurons fire correlated action potentials at much higher frequencies driven by slower, smaller, and broadly synchronized depolarization.


Mild hyperhomocysteinemia is not associated with cardiac allograft coronary disease.
Hyperhomocysteinemia is an independent risk factor for coronary disease and elevated plasma homocysteine levels have been documented in heart transplant recipients. The aim of this study was to test the hypothesis that homocysteine levels are associated with presence or absence of transplant coronary artery disease. Forty-three non-smoking adults were recruited, all of whom had received a heart transplant between 2 and 7 yr previously. All 43 had blood drawn for fasting homocysteine level on the day of presentation. All patients had undergone diagnostic coronary angiography within the past 6 months. For all patients, the average fasting plasma homocysteine level was 17.0+/-SD 6.6 micromol/L with a range from 6.0 to 36.9 micromol/L. Twenty-six patients (60%) had fasting plasma homocysteine levels above 15.0 micromol/L. On the basis of arteriography, patients were categorized as those with angiographically normal (n=22) or abnormal (n=21) coronary arteries. There was no difference in the mean plasma homocysteine level comparing patients with angiographically normal (17.2+/-SD 7.0 micromol/L) to those with abnormal (16.8+/-SD 6.2 micromol/L) coronary arteries. Plasma homocysteine levels increased with increasing plasma creatinine levels (r=0.63, p<0.0001) and with decreasing vitamin B6 levels (r=-0.56, p<0.0001). Mild hyperhomocysteinemia is a consistent finding among heart transplant recipients. This finding was not associated with transplant coronary artery disease in our patients. The combination of renal dysfunction and vitamin B6 deficiency may explain the unusual prevalence of hyperhomocysteinemia in heart transplant recipients.


Comprehensive identification of proteins from MALDI imaging.
Matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) is a powerful tool for the visualization of proteins in tissues and has demonstrated considerable diagnostic and prognostic value. One main challenge is that the molecular identity of such potential biomarkers mostly remains unknown. We introduce a generic method that removes this issue by systematically identifying the proteins embedded in the MALDI matrix using a combination of bottom-up and top-down proteomics. The analyses of ten human tissues lead to the identification of 1400 abundant and soluble proteins constituting the set of proteins detectable by MALDI IMS including >90% of all IMS biomarkers reported in the literature. Top-down analysis of the matrix proteome identified 124 mostly N- and C-terminally fragmented proteins indicating considerable protein processing activity in tissues. All protein identification data from this study as well as the IMS literature has been deposited into MaTisse, a new publically available database, which we anticipate will become a valuable resource for the IMS community.


GeneTrees: a phylogenomics resource for prokaryotes.
The GeneTrees phylogenomics system pursues comparative genomic analyses from the perspective of gene phylogenies for individual genes. The GeneTrees project has the goal of providing detailed evolutionary models for all protein-coding gene components of the fully sequenced genomes. Currently, a database of alignments and trees for all protein sequences for 325 fully sequenced and annotated prokaryote genomes is available. The prokaryote database contains 890,000 protein sequences organized into over 100,000 alignments, each described by a phylogenetic tree. An original homology group discovery tool assembles sets of related proteins from all versus all pairwise alignments. Multiple alignments for each homology group are stored and subjected to phylogenetic tree inference. A graphical web interface provides visual exploration of the GeneTrees database. Homology groups can be queried by sequence identifiers or annotation terms. Genomes can be browsed visually on a gene map of each chromosome or plasmid. Phylogenetic trees with support values are displayed in conjunction with the associated sequence alignment. A variety of classes of information can be selected to label the tree tips to aid in visual evaluation of annotation and gene function. This web interface is available at http://genetrees.vbi.vt.edu.


NFAT restricts osteochondroma formation from entheseal progenitors.
Osteochondromas are common benign osteocartilaginous tumors in children and adolescents characterized by cartilage-capped bony projections on the surface of bones. These tumors often cause pain, deformity, fracture, and musculoskeletal dysfunction, and they occasionally undergo malignant transformation. The pathogenesis of osteochondromas remains poorly understood. Here, we demonstrate that nuclear factor of activated T cells c1 and c2 (NFATc1 and NFATc2) suppress osteochondromagenesis through individual and combinatorial mechanisms. In mice, conditional deletion of NFATc1 in mesenchymal limb progenitors, Scleraxis-expressing (Scx-expressing) tendoligamentous cells, or postnatally in Aggrecan-expressing cells resulted in osteochondroma formation at entheses, the insertion sites of ligaments and tendons onto bone. Combinatorial deletion of NFATc1 and NFATc2 gave rise to larger and more numerous osteochondromas in inverse proportion to gene dosage. A population of entheseal NFATc1- and Aggrecan-expressing cells was identified as the osteochondroma precursor, previously believed to be growth plate derived or perichondrium derived. Mechanistically, we show that NFATc1 restricts the proliferation and chondrogenesis of osteochondroma precursors. In contrast, NFATc2 preferentially inhibits chondrocyte hypertrophy and osteogenesis. Together, our findings identify and characterize a mechanism of osteochondroma formation and suggest that regulating NFAT activity is a new therapeutic approach for skeletal diseases characterized by defective or exaggerated osteochondral growth.


Probing the dynamic nature of signalling pathways by IMAC and SELDI-tof MS.
One major obstacle to the analysis of signalling pathways is the dynamic nature of signalling response to environmental stimuli. To overcome this limitation we applied immobilized metal affinity chromatography (IMAC) in combination with SELDI-tof MS to investigate the temporal variation of protein phosphorylation. We analysed the phospho-proteome variations in our model organism, Leishmania donovani, in response to changes in pH and temperature, which induce differentiation from promastigotes to amastigotes. Investigation of total cell extracts did not allow promastigote and amastigote life cycle stages to be distinguished. However, using IMAC enriched samples, the pattern and intensity of phospho-proteins analysed distinguished both stages reproducibly. Approximately 61% of the phospho-proteins analysed were significantly different in abundance (p<0.02). Of these 61%, 73% showed an increased phosphorylation in promastigotes while 27% showed an increase phosphorylation in amastigotes. The workflow developed is currently being applied to the temporal analysis of environmental stimuli.


5-HT modulation of auditory and visual sensorimotor gating: II. Effects of the 5-HT2A antagonist MDL 100,907 on disruption of sound and light prepulse inhibition produced by 5-HT agonists in Wistar rats.
Increasing evidence suggests an important role of 5-HT, and 5-HT2A receptors in particular, in the etiology and treatment of schizophrenia. The prepulse inhibition paradigm is used as a model for sensorimotor gating processes that are disrupted in schizophrenia. The present study used the selective serotonin2A (5-HT2A) antagonist and putative antipsychotic agent MDL 100,907 to evaluate the contribution of 5-HT2A receptors to the disruptions of prepulse inhibition produced by several 5-HT agonists. The D2 antagonist haloperidol was used to evaluate a possible interaction with dopamine neurons. Sound or light prepulses were used to measure the generality of these drug effects on cross-modal prepulse inhibition. In the first study, MDL 100,907 antagonized the disruptions of auditory prepulse inhibition produced by the 5-HT releasing agents fenfluramine and 3,4-methylenedioxymethamphetamine (MDMA). These effects on prepulse inhibition were modality-specific in that MDL 100,907 did not reverse the effects of the 5-HT releasers on visual prepulse inhibition. Haloperidol did not alter the disruptive effects of MDMA or fenfluramine on either auditory or visual prepulse inhibition. In the second study, the direct acting 5-HT2A/2C receptor agonist/hallucinogen (+)1-4-iodo-2,5-dimethoxyphenyl-2-aminopropane (DOI) consistently disrupted auditory prepulse inhibition, and this effect was blocked by MDL 100,907 but not by haloperidol. A dose-response analysis demonstrated that MDL 100,907 potently antagonized DOI disrupted auditory prepulse inhibition, with an ED50 of 0.04 mg/kg, IP. DOI did not consistently disrupt visual prepulse inhibition. In summary, these data indicate that, at least under the conditions of the present studies, the disruptions of auditory prepulse inhibition produced by fenfluramine, MDMA, and DOI result from stimulation of 5-HT2A receptors. Furthermore, these disruptions do not involve direct or indirect stimulation of D2 receptors. The identity of the 5-HT receptor(s) underlying the disruptive effects of fenfluramine or MDMA on visual prepulse inhibition has not yet been identified. MDL 100,907 may be generally useful in CNS disorders in which excessive 5-HT2A receptor tone disrupts sensory gating processes.


Optimistic update bias holds firm: Three tests of robustness following Shah et al.
A diverse body of research has demonstrated that people update their beliefs to a greater extent when receiving good news compared to bad news. Recently, a paper by Shah et al. claimed that this asymmetry does not exist. Here we carefully examine the experiments and simulations described in Shah et al. and follow their analytic approach on our data sets. After correcting for confounds we identify in the experiments of Shah et al., an optimistic update bias for positive life events is revealed. Contrary to claims made by Shah et al., we observe that participants update their beliefs in a more Bayesian manner after receiving good news than bad. Finally, we show that the parameters Shah et al. pre-selected for simulations are at odds with participants' data, making these simulations irrelevant to the question asked. Together this report makes a strong case for a true optimistic asymmetry in belief updating.


Knowledge and causal attributions for mental disorders in HIV-positive children and adolescents: results from rural and urban Uganda.
Increasing availability of antiretroviral treatment (ART) has led HIV to be considered a chronic disease, shifting attention to focus on quality of life including mental wellbeing. We investigated knowledge and causal attributions for mental disorders in HIV-positive children and adolescents in rural and urban Uganda. This qualitative study was nested in an epidemiological mental health study among HIV-positive children and adolescents aged 5-17 years in rural and urban Uganda. In-depth interviews were conducted with caregivers of HIV-positive children (5-11 years) and adolescents (12-17 years) in HIV care. Interviews were audio recorded with permission from participants and written consent and assent sought before study procedures. Thirty eight participants (19 caregivers, 19 children/adolescents) were interviewed. Age range of caregivers was 28-69 years; majority were female (17). Caregivers had little knowledge on mental disorders ;only 3 related the vignette to a mental problem and attributed it to: improper upbringing, violence, poverty and bereavement. Five adolescents identified vignettes as portraying mental disorders caused by: ill-health of parents, bereavement, child abuse, discrimination, HIV and poverty. Caregivers are not knowledgeable about behavioural and emotional challenges in HIV-positive children/adolescents. Mental health literacy programmes at HIV care clinics are essential to enhance treatment-seeking for mental health.


Exposure to dioxin and nonneoplastic mortality in the expanded IARC international cohort study of phenoxy herbicide and chlorophenol production workers and sprayers.
The authors studied noncancer mortality among phenoxyacid herbicide and chlorophenol production workers and sprayers included in an international study comprising 36 cohorts from 12 countries followed from 1939 to 1992. Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin or higher chlorinated dioxins (TCDD/HCD) was discerned from job records and company questionnaires with validation by biologic and environmental measures. Standard mortality ratio analyses suggested a moderate healthy worker effect for all circulatory diseases, especially ischemic heart disease, among both those exposed and those not exposed to TCDD/HCD. In Poisson regression analyses, exposure to TCDD/HCD was not associated with increased mortality from cerebrovascular disease. However, an increased risk for circulatory disease, especially ischemic heart disease (rate ratio [RR] 1.67, 95% confidence interval [Cl] 1.23-2.26) and possibly diabetes (RR 2.25, 95% Cl 0.53-9.50), was present among TCDD/HCD-exposed workers. Risks tended to be higher 10 to 19 years after first exposure and for those exposed for a duration of 10 to 19 years. Mortality from suicide was comparable to that for the general population for all workers exposed to herbicides or chlorophenols and was associated with short latency and duration of exposure. More refined investigations of the ischemic heart disease and TCDD/HCD exposure association are warranted.


The Significance of Autoantibody Changes Over Time in Primary Biliary Cirrhosis.
In primary biliary cirrhosis (PBC), the antimitochondrial antibody is a cornerstone of diagnosis, but there have been conflicting reports about the correlation of autoantibodies with disease stage and prognosis. We studied whether autoantibody levels changed over time and sought correlations with clinical outcomes in a cohort of patients with PBC. We tested serial serum samples from patients with PBC at a research institution for several autoantibodies. Long-term clinical follow-up data were used to calculate the slopes (change over time) for autoantibodies, platelet count, Ishak fibrosis score, biopsy copper, and number of portal areas with bile ducts. An adverse clinical outcome was defined as hepatic decompensation, development of hepatocellular carcinoma, liver transplantation, or liver-related death. We performed linear or logistic regression or Fisher exact test as appropriate, adjusting for multiple comparisons. Twenty-seven patients with PBC with 145 serum samples were studied. Of the cohort, 85% was white, 81% was female, and median follow-up time was 20 years. Of the autoantibodies tested, only sp100 changed significantly over time. The sp100 slope was inversely associated with the Ishak fibrosis slope (parameter estimate, -0.05; P = .0003). While changes in most autoantibodies over time do not seem to correlate with clinical outcomes in PBC, a change in the sp100 autoantibody level may have prognostic utility with respect to the development of fibrosis on liver biopsy.


Peeling single-stranded DNA from graphite surface to determine oligonucleotide binding energy by force spectroscopy.
We measured the force required to peel single-stranded DNA molecules from single-crystal graphite using chemical force microscopy. Force traces during retraction of a tip chemically modified with oligonucleotides displayed characteristic plateaus with abrupt force jumps, which we interpreted as a steady state peeling process punctuated by complete detachment of one or more molecules. We were able to differentiate between bases in pyrimidine homopolymers; peeling forces were 85.3 - 4.7 pN for polythymine and 60.8 +/- 5.5 pN for polycytosine, substantially independent of salt concentration and the rate of detachment. We developed a model for peeling a freely jointed chain from the graphite surface and estimated the average binding energy per monomer to be 11.5 +/- 0.6 k(B)T and 8.3 +/- 0.7 k(B)T in the cases of thymine and cytosine nucleotides, respectively. The equilibrium free-energy profile simulated using molecular dynamics had a potential well of 18.9 k(B)T for thymidine, showing that nonelectrostatic interactions dominate the binding. The discrepancy between the experiment and theory indicates that not all bases are adsorbed on the surface or that there is a population of conformations in which they adsorb. Force spectroscopy using oligonucleotides covalently linked to AFM tips provides a flexible and unambiguous means to quantify the strength of interactions between DNA and a number of substrates, potentially including nanomaterials such as carbon nanotubes.


Food Store Environment Modifies Intervention Effect on Fruit and Vegetable Intake among Low-Income Women in North Carolina.
Background. The aim of the study is to determine how the food store environment modifies the effects of an intervention on diet among low-income women. Study Design. A 16-week face-to-face behavioral weight loss intervention was delivered among low income midlife women. Methods. The retail food environment for all women was characterized by (1) the number and type of food stores within census tracts; (2) availability of healthy foods in stores where participants shop; (3) an aggregate score of self-reported availability of healthy foods in neighborhood and food stores. Statistical Analyses. Multivariable linear regression was used to model the food store environment as an effect modifier between the intervention effect of fruit and vegetable serving change. Results. Among intervention participants with a low perception of availability of healthy foods in stores, the intervention effect on fruit and vegetable serving change was greater [1.89, 95% CI (0.48, 3.31)] compared to controls. Among intervention participants residing in neighborhoods with few super markets, the intervention effect on fruit and vegetable serving change was greater [1.62, 95% CI (1.27, 1.96)] compared to controls. Conclusion. Results point to how the food store environment may modify the success of an intervention on diet change among low-income women.


Phosphatidylinositol 3-kinase (PI3K) signaling via glycogen synthase kinase-3 (Gsk-3) regulates DNA methylation of imprinted loci.
Glycogen synthase kinase-3 (Gsk-3) isoforms, Gsk-3α and Gsk-3β, are constitutively active, largely inhibitory kinases involved in signal transduction. Underscoring their biological significance, altered Gsk-3 activity has been implicated in diabetes, Alzheimer disease, schizophrenia, and bipolar disorder. Here, we demonstrate that deletion of both Gsk-3α and Gsk-3β in mouse embryonic stem cells results in reduced expression of the de novo DNA methyltransferase Dnmt3a2, causing misexpression of the imprinted genes Igf2, H19, and Igf2r and hypomethylation of their corresponding imprinted control regions. Treatment of wild-type embryonic stem cells and neural stem cells with the Gsk-3 inhibitor, lithium, phenocopies the DNA hypomethylation at these imprinted loci. We show that inhibition of Gsk-3 by phosphatidylinositol 3-kinase (PI3K)-mediated activation of Akt also results in reduced DNA methylation at these imprinted loci. Finally, we find that N-Myc is a potent Gsk-3-dependent regulator of Dnmt3a2 expression. In summary, we have identified a signal transduction pathway that is capable of altering the DNA methylation of imprinted loci.


Relationship between the debrisoquine hydroxylase polymorphism and cancer susceptibility.
There have been a series of reports on the association of a genetic polymorphism at the cytochrome P450 CYP2D6 gene locus with cancer susceptibility. Many of these reports have remained contradictory either because of small numbers of patients studied or because of the limitations and controversy surrounding the pharmacokinetic assay used to identify affected individuals (poor metabolizers; PMs). We have recently developed a DNA-based assay that will allow the unequivocal identification of poor metabolizers and have applied this to the study of 1635 patients with different forms of cancer. Out of 361 lung cancer patients studied no statistically significant change in the proportion of PMs relative to controls was found. However, a significant increase in the proportion of poor metabolizers or heterozygotes was seen in leukaemia, bladder cancer and melanoma patients. This could be explained by a role for CYP2D6 in carcinogen detoxification or by linkage to another cancer-causing gene.


Gaming the system. Dodging the rules, ruling the dodgers.
Although traditional obligations of fidelity require physicians to deliver quality care to their patients, including to utilize costly technologies, physicians are steadily losing their accustomed control over the necessary resources. The "economic agents" who own the medical and monetary resources of care now impose a wide array of rules and restrictions in order to contain their costs of operation. However, physicians can still control resources indirectly through "gaming the system," employing tactics such as "fudging" that exploit resource rules' ambiguity and flexibility to bypass the rules while ostensibly honoring them. Physicians may be especially inclined to game the system where resource rules seriously underserve patients' needs, where economic agents seem to be "gaming the patient," with needless obstacles to care, or where others, such as hospitals or even physicians themselves, may be denied needed reimbursements. Though tempting, gaming is morally and medically hazardous. It can harm patients and society, offend honesty, and violate basic principles of contractual and distributive justice. It is also, in fact, usually unnecessary in securing needed resources for patients. More fundamentally, we must reconsider what physicians owe their patients. They owe what is theirs to give: their competence, care and loyalty. In light of medicine's changing economics, two new duties emerge: economic advising, whereby physicians explicitly discuss the economic as well as medical aspects of each treatment option; and economic advocacy, whereby physicians intercede actively on their patients' behalf with the economic agents who control the resources.


Invasive fungal infections following liver transplantation - risk factors, incidence and outcome.
The incidence of invasive fungal infections (IFIs), particularly candidiasis and aspergillosis, following solid organ transplantation vary from 1.4% to 42%. IFIs most commonly occur after orthotropic liver transplantation (OLT), lung/heart and pancreas transplantation. Mortality related to IFIs depends on the type of transplant and vary from 3% to 100% of cases. The results largely depend on early initiation of specific treatment for IFIs. Therefore, the diagnosis has to be prompt and based on known risk factors - time of surgical procedure, type of biliary anastomosis, blood loss, rate of rejection and re-transplantation. We evaluated the incidence of fungal infections in patients after liver transplantation in our center. The retrospective analysis of 175 consecutive OLT patients was undertaken to estimate incidence, risk factors and clinical courses of IFIs in the last 6 years at our center. Infections involving Aspergillus (6 cases), Candida (24 cases) and Cryptococcus (1 case) were observed in 17.7% of our recipients. Except for 1 case (Cryptococcus encephalitis), all of the episodes developed during the first month post-transplantation. All cases of lung aspergillosis developed in patients with autoimmune cholestasis prior to transplantation. In 1 case after transplantation, in a patient with bile duct necrosis requiring reoperation, pneumonia developed. In 3 cases, pulses of steroids were used to treat acute rejection. Apart from that, none of the potential risk factors of IFIs described by other authors were noted. Five out of 6 cases of aspergillosis survived on combined antifungal therapy. The recipient diagnosed with cryptococcal encephalitis died. All cases with urinary tract (n=18; 8.6%) or respiratory (n=6; 3.4%) candidiasis survived. Early diagnosis and prompt treatment is fundamental for patient survival.


Developmental mechanisms for the generation of telencephalic interneurons.
Interneurons, which release the neurotransmitter γ-aminobutyric acid (GABA), are the major inhibitory cells of the central nervous system (CNS). Despite comprising only 20-30% of the cerebral cortical neuronal population, these cells play an essential and powerful role in modulating the electrical activity of the excitatory pyramidal cells onto which they synapse. Although interneurons are present in all regions of the mature telencephalon, during embryogenesis these cells are generated in specific compartments of the ventral (subpallial) telencephalon known as ganglionic eminences. To reach their final destinations in the mature brain, immature interneurons migrate from the ganglionic eminences to developing telencephalic structures that are both near and far from their site of origin. The specification and migration of these cells is a complex but precisely orchestrated process that is regulated by a combination of intrinsic and extrinsic signals. The final outcome of which is the wiring together of excitatory and inhibitory neurons that were born in separate regions of the developing telencephalon. Disruption of any aspect of this sequence of events during development, either from an environmental insult or due to genetic mutations, can have devastating consequences on normal brain function.


A readiness assessment for the prevention of alcohol-related harm in West Africa: A new methodological approach to inform practice and policy.
Alcohol-related harm is a growing concern globally and particularly in West Africa. However, tools for assessing the readiness for prevention of alcohol-related harm in low-resource settings have been lacking. We modified the WHO tool, the Readiness Assessment for the Prevention of Child Maltreatment Short Form (RAP-CM), to assess readiness for the prevention of alcohol-related harm across West Africa. We conducted a cross-sectional survey in the fall of 2020, distributed by the West Africa Alcohol Policy Alliance to their member alliances and stakeholders, predominantly community-based organizations (CBOs) and non-governmental organizations (NGOs), across 7 countries in West Africa (N = 140). The survey included modified measures from the RAP-CM short form. In terms of general readiness, the overall adjusted aggregate score for West Africa was 45.0% (ranging from 42.9% in Liberia to 52.7% in Senegal). Of the ten domains assessed (on a 0-10 scale), across all countries, knowledge of alcohol-related harm prevention (8.3) and legislation, mandates, and policies (6.7) received the highest readiness scores. The lowest readiness scores were observed for human and technical resources (2.5), attitudes toward preventing alcohol-related harm (2.7), and the will to address the problem (2.9). Our results demonstrate substantial variability across domains in the readiness to address alcohol-related harm with clear strengths and limitations for future priority setting and capacity building. The barriers to progress include attitudes toward alcohol-related harm prevention, lack of willingness to address the problem, and limited human and technical resources available. These barriers need to be mitigated to address the high burden of alcohol-related harm in the region and to inform both practice and policy.


Aspiration after stroke: lesion analysis by brain MRI.
Aspiration is a common problem following stroke, resulting in feeding difficulties and aspiration pneumonia. Despite past studies using clinical assessments and computed tomographic (CT) scans of the head, the correlation of stroke location with aspiration remains unclear. Since brain magnetic resonance imaging is more sensitive than CT for many stroke types, we have correlated MRI lesions with aspiration in patients who have sustained a stroke. We selected patients with acute stroke who underwent brain MRI and a swallowing evaluation. Aspiration was present in 21 of 38 patients (55%). Patients with just small vessel infarcts had a significantly lower occurrence of aspiration (3 of 14, 21%) compared to those with both large- and small-vessel infarcts (15 of 20, 75%, p = 0.002). Multivariate analysis of several specific brain areas failed to identify a significant association between stroke location and the occurrence of aspiration. These findings suggest that patients who have experienced stroke should be individually evaluated for swallowing dysfunction regardless of stroke location or size, since even small-vessel strokes can be associated with aspiration in greater than 20% of cases.


Analysis of gene expression and chemoresistance of CD133+ cancer stem cells in glioblastoma.
Recently, a small population of cancer stem cells in adult and pediatric brain tumors has been identified. Some evidence has suggested that CD133 is a marker for a subset of leukemia and glioblastoma cancer stem cells. Especially, CD133 positive cells isolated from human glioblastoma may initiate tumors and represent novel targets for therapeutics. The gene expression and the drug resistance property of CD133 positive cancer stem cells, however, are still unknown. In this study, by FACS analysis we determined the percentage of CD133 positive cells in three primary cultured cell lines established from glioblastoma patients 10.2%, 69.7% and 27.5%, respectively. We also determined the average mRNA levels of markers associated with neural precursors. For example, CD90, CD44, CXCR4, Nestin, Msi1 and MELK mRNA on CD133 positive cells increased to 15.6, 5.7, 337.8, 21.4, 84 and 1351 times, respectively, compared to autologous CD133 negative cells derived from cell line No. 66. Additionally, CD133 positive cells express higher levels of BCRP1 and MGMT mRNA, as well as higher mRNA levels of genes that inhibit apoptosis. Furthermore, CD133 positive cells were significantly resistant to chemotherapeutic agents including temozolomide, carboplatin, paclitaxel (Taxol) and etoposide (VP16) compared to autologous CD133 negative cells. Finally, CD133 expression was significantly higher in recurrent GBM tissue obtained from five patients as compared to their respective newly diagnosed tumors. Our study for the first time provided evidence that CD133 positive cancer stem cells display strong capability on tumor's resistance to chemotherapy. This resistance is probably contributed by the CD133 positive cell with higher expression of on BCRP1 and MGMT, as well as the anti-apoptosis protein and inhibitors of apoptosis protein families. Future treatment should target this small population of CD133 positive cancer stem cells in tumors to improve the survival of brain tumor patients.


The value of an evidence database for occupational therapists: an international online survey.
Online evidence databases can provide access to high quality evidence at the point of care, making evidence-based practice more achievable. A discipline-specific online bibliographic database called OTseeker (www.otseeker.com) was designed for use by occupational therapists. The database is free, and contains citations and abstracts of systematic reviews, and critically appraised randomized controlled trials relevant to occupational therapy. The aim of this study was to investigate search practices of database users, their views on its functionality, and the reported impact, if any, on their practice from using OTseeker. An online survey, placed on the database website for 30 days. Potential participants were users of OTseeker during a 30-day period. A total of 498 people who had used the database more than once from over 40 countries completed the survey. Three hundred and nine (62%) participants believed that OTseeker had improved their ability to locate research about the effectiveness of occupational therapy interventions, and 92 (19%) agreed that the information in the database had contributed to a change in practice. Those reporting no practice changes agreed that use of OTseeker had improved their knowledge generally (n=189; 38%), confirmed their practice (n=75; 15%), or revealed that there was insufficient research relevant to their search topic (n=92; 19%). Features of the database which helped respondents locate research evidence included: having discipline-specific content, providing critical appraisal ratings for randomized controlled trials, and presenting search results ranked for methodological quality. This study confirms the value of a discipline-specific, online database for helping occupational therapists locate high quality research evidence. Information located on databases such as OTseeker can help change or confirm practice, and improve knowledge.


Histopathologic predictors of pilocytic astrocytoma event-free survival.
Pilocytic astrocytoma (PA) is the most common pediatric brain tumor. Most arise in the cerebellum, but they also can develop in the brainstem and optic nerve, where gross total resection (GTR) is not possible. In the absence of GTR, significant variability in both clinical behavior and histology exists. To identify potential markers associated with poor clinical outcome, we retrospectively assessed pathological features in 107 patients with PAs. We identified four pathological features (necrosis, oligodendroglioma-like features, vascular hyalinization, and calcification) that showed a significant correlation with decreased event-free survival (EFS). Similar to previous reports, we also found that PAs involving the optic pathway were associated with worse EFS compared with those arising in other locations. In contrast, mitotic index, p53 immunoreactivity and hyperactivation of several mitogenic signaling pathways (MAPK, CREB, mTOR) did not demonstrate a statistically significant relationship with EFS. Lastly, we did find a statistical trend between EFS and the number of CD68+ cells, suggesting that non-neoplastic elements of the tumor microenvironment may influence subsequent growth and clinical recurrence. Collectively, the identification of specific histopathologic features associated with clinical outcome may improve our ability to determine which PAs are more likely to exhibit clinical progression and require more vigilant observation.


Effects of age and parity on mammary gland lesions and progenitor cells in the FVB/N-RC mice.
The FVB/N mouse strain is extensively used in the development of animal models for breast cancer research. Recently it has been reported that the aging FVB/N mice develop spontaneous mammary lesions and tumors accompanied with abnormalities in the pituitary glands. These observations have a great impact on the mouse models of human breast cancer. We have developed a population of inbred FVB/N mice (designated FVB/N-RC) that have been genetically isolated for 20 years. To study the effects of age and parity on abnormalities of the mammary glands of FVB/N-RC mice, twenty-five nulliparous and multiparous (3-4 pregnancies) females were euthanized at 16-22 months of age. Examination of the mammary glands did not reveal macroscopic evidence of mammary gland tumors in either aged-nulliparous or multiparous FVB/N-RC mice (0/25). However, histological analysis of the mammary glands showed rare focal nodules of squamous changes in 2 of the aged multiparous mice. Mammary gland hyperplasia was detected in 8% and 71% of the aged-nulliparous and aged-multiparous mice, respectively. Epithelial contents and serum levels of triiodothyronine were significantly higher in the experimental groups than the 14-wk-old control mice. Immuno-histochemical staining of the pituitary gland pars distalis showed no difference in prolactin staining between the control and the aged mice. Tissue transplant and dilution studies showed no effect of age and/or parity on the ability of putative progenitor cells present among the injected mammary cells to repopulate a cleared fat pad and develop a full mammary gland outgrowth. This FVB/N-RC mouse substrain is suitable to develop mouse models for breast cancer.


Challenges and prospects in genome-wide quantitative trait loci mapping of standing genetic variation in natural populations.
A considerable challenge in evolutionary genetics is to understand the genetic mechanisms that facilitate or impede evolutionary adaptation in natural populations. For this, we must understand the genetic loci contributing to trait variation and the selective forces acting on them. The decreased costs and increased feasibility of obtaining genotypic data on a large number of individuals have greatly facilitated gene mapping in natural populations, particularly because organisms whose genetics have been historically difficult to study are now within reach. Here we review the methods available to evolutionary ecologists interested in dissecting the genetic basis of traits in natural populations. Our focus lies on standing genetic variation in outbred populations. We present an overview of the current state of research in the field, covering studies on both plants and animals. We also draw attention to particular challenges associated with the discovery of quantitative trait loci and discuss parallels to studies on crops, livestock, and humans. Finally, we point to some likely future developments in genetic mapping studies.


Can chronic neuropathic pain following thoracic surgery be predicted during the postoperative period?
Chronic pain following thoracic surgery is common and associated with neuropathic symptoms, however, the proportion of patients with neuropathic pain in the immediate postoperative period is unknown. We aimed to determine the proportion of patients who have neuropathic symptoms and signs immediately after, and at three months following thoracic surgery. The study was designed as a prospective observational cohort study. We identified patients with pain of predominantly neuropathic origin using the Leeds Assessment of Neuropathic Symptoms and Signs (LANSS) score in the immediate postoperative period and the self-report LANSS (S-LANSS) version three months after surgery. One hundred patients undergoing video assisted thoracic surgery (VATS) or thoracotomy completed LANSS scores preoperatively and in the immediate postoperative period. Eighty-seven percent completed three months S-LANSS follow-up scores. Eight percent of patients had positive LANSS scores in the immediate postoperative period; 22% of patients had positive S-LANSS scores three months following surgery. There was a significant association between positive scores in the acute and chronic periods (relative risk (RR) 3.5, [95% confidence interval (CI) 1.7-7.2]). Identifying pain of predominantly neuropathic origin in the postoperative period with a simple pain score can help identify those at risk of developing chronic pain with these features following thoracic surgery.


Parental stress, pediatric quality of life, and behavior at baseline and one-year follow-up: Results from the FEBSTAT study.
Febrile status epilepticus is a serious and frightening event in the life of the child and parent. It is regarded as a medical emergency with potential long lasting consequences. The purpose of this study was to look at the immediate and long term effects of such an event on parental stress and parents' perception of their child's physical and psychosocial wellbeing. From 2003 to 2010, 199 subjects, age 1 month to 5 years, were recruited as part of a prospective, multicenter study (FEBSTAT) of consequences of febrile status epilepticus (FSE). At one month and one year after the episode of FSE, parents were asked to complete the Parenting Stress Index, short form (PSI/SF), the Pediatric Quality of Life Inventory (PedsQL) and the Child Behavior Checklist (CBCL). In addition to PedsQL and CBCL in the FEBSTAT subjects only, a comparison was made between Columbia Study of First Febrile Seizures subjects with a first simple febrile seizure (SFS) and the FEBSTAT group, including 15 subjects with FSE from the Columbia group, in the area of parental stress which was administered at the same time intervals in both studies. At baseline, the PSI/SF was statistically significantly higher for SFS versus FSE on the parent-child dysfunctional score and the total raw score, however at one year this difference resolved. In the FSE group, significantly higher parental stress over one year was reported in children with abnormal versus normal prior development (p= 0.02). Prior abnormal development was a risk factor at 1 year for lower total PEDSQL (p=0.01) versus prior normal development. Mean scores on the CBCL at baseline and 1 year were within the normal range for both empirically based scales and major risk factors. Parents of children experiencing a SFS experienced more stress at baseline than those with FSE. Families of children in the FEBSTAT cohort with identified development problems at baseline that continued, or progressed over the one year period, reported decreasing QOL.


A 13-Year-Old Boy Who Has Kawasaki Disease Shock Syndrome Presents with Parotitis.
We report a 13-year-old boy who (initially) had symptoms of toxic shock-like syndrome and mumps. Then, the patient was hospitalized in the pediatric intensive care unit (PICU) because of his ongoing hemodynamic instability (low blood pressure of 70/30 mm Hg and capillary refill time of > 4 seconds). During his stay in the PICU, the patient was treated with fluid resuscitation and vasoactive infusion and at the same time was diagnosed with Kawasaki disease shock syndrome (KDSS), when giant right coronary artery aneurysms were detected on echocardiographic examination. This case illustrates the risk of KDSS in patient who carries both parotitis and toxic shock-like syndrome. The clinicians should be cautious about detecting any types of coronary artery aneurysms in such patients. This is the first case of KDSS associated with parotitis reported in the literature.


Transcranial Doppler and Hematoma Expansion in Acute Spontaneous Primary Intracerebral Hemorrhage.
The data on the role of Transcranial Doppler (TCD) in the management of acute primary intracerebral hemorrhage (ICH) is meager. To study TCD variables associated with hematoma expansion in acute primary ICH. The study was carried out in the neurosciences department of Post Graduate Institute of Medical Education and Research (PGIMER), Chandigarh from July 2010 to September 2011 employing a prospective, double blinded non randomized study design. Acute ICH patients within 24 h of symptom onset were recruited. Baseline neuroimaging study (Computerized tomography, CT scan of brain) was performed to assess the pure hematoma volume by AXBXC/2 method. Baseline TCD parameters were obtained from both the middle cerebral arteries (MCAs; affected and unaffected hemisphere): Peak Systolic velocity, End Diastolic velocity, Mean Flow velocity, Resistance Index, and Pulsatility Index. Follow up (24 h) assessment of hematoma volume and TCD were carried out. Each of the TCD variables were compared in hematoma expansion (>33% increase in hematoma volume on the follow-up CT) and non-expansion group. On univariate analysis, the Student's t-test and contingency tables with the X2 test were used. A forward stepwise multivariate logistic regression analysis with hematoma expansion at 24 h as the dependent variable and ROC analysis was carried out, using SPSS software version 16 (Chicago, IL). P value < 0.05 was considered significant. Twenty-five patients completed the study. Ten patients (40%) had hematoma expansion. Multivariate analysis revealed unaffected hemisphere MCA Pulsatility Index ratio [unaffected hemisphere MCA Follow up Pulsatility Index/baseline Pulsatility Index] of > 1.055 as the lone correlate of hematoma expansion (sensitivity of 90% and specificity of 60%). Frequent assessment with TCD could aid in prediction of hematoma expansion by measuring unaffected hemisphere Pulsatility Index ratios.


Sarcopenia and Health Care Utilization in Older Women.
Although there are several consensus definitions of sarcopenia, their association with health care utilization has not been studied. We included women from the prospective Study of Osteoporotic Fractures with complete assessment of sarcopenia by several definitions at the Study of Osteoporotic Fractures Year 10 (Y10) exam (1997-1998) who also had available data from Medicare Fee- For-Service Claims (N = 566) or Kaiser Encounter data (N = 194). Sarcopenia definitions evaluated were: International Working Group, European Working Group for Sarcopenia in Older Persons, Foundation for the NIH Sarcopenia Project, Baumgartner, and Newman. Hurdle models and logistic regression were used to assess the relation between sarcopenia status (the summary definition and the components of slowness, weakness and/or lean mass) and outcomes that included hospitalizations, cumulative inpatient days/year, short-term (part A paid) skilled nursing facility stay in the 3 years following the Y10 visit. None of the consensus definitions, nor the definition components of weakness or low lean mass, was associated with increased risk of hospitalization or greater likelihood of short-term skilled nursing facility stay. Women with slowness by any criterion definition were about 50% more likely to be hospitalized; had a greater rate of hospitalization days amongst those hospitalized; and had 1.8 to 2.1 times greater likelihood of a short-term skilled nursing facility stay than women without slowness. There was the suggestion of a protective association of low lean mass by the various criterion definitions on short-term skilled nursing facility stay. Estimated effects of sarcopenia on health care utilization were negligible. However, slowness was associated with greater health care utilization.


The clinical relevance of pharmacological blood pressure lowering mechanisms.
Hypertension is a major risk factor for stroke, myocardial infarction, heart failure and kidney failure. Clinical trials demonstrate that lowering blood pressure can substantially reduce cardiovascular risk, and current clinical practice guidelines identify lowering blood pressure as a priority in the treatment of people with hypertension and set clear treatment targets. In addition to lifestyle modifications, which should be pursued in all patients, a large number of antihypertensive drugs and combinations of such drugs are part of the modern treatment armamentarium. Following the publication of the Antihypertensive and Lipid-Lowering Treatment to prevent Heart Attack Trial (ALLHAT), some investigators and clinicians have suggested that thiazide diuretics should be the first drug of choice for the treatment of all hypertensive patients. However, antihypertensive drugs differ greatly in their mechanisms of action, pharmacokinetic properties and side effects. There are also large variations among hypertensive patients with regards to the pathophysiology of their blood pressure elevation, response to individual antihypertensive drugs, comorbidities and propensity to drug-related side effects. Therefore, a uniform formula for the treatment of hypertension is not likely to achieve optimal results. Rather, treatment strategies based on careful, comprehensive and individualized patient assessments should be pursued.


Noncovalent cell surface engineering: incorporation of bioactive synthetic glycopolymers into cellular membranes.
The controlled addition of structurally defined components to live cell membranes can facilitate the molecular level analysis of cell surface phenomena. Here we demonstrate that cell surfaces can be engineered to display synthetic bioactive polymers at defined densities by exogenous membrane insertion. The polymers were designed to mimic native cell-surface mucin glycoproteins, which are defined by their dense glycosylation patterns and rod-like structures. End-functionalization with a hydrophobic anchor permitted incorporation into the membranes of live cultured cells. We probed the dynamic behavior of cell-bound glycopolymers bearing various hydrophobic anchors and glycan structures using fluorescence correlation spectroscopy (FCS). Their diffusion properties mirrored those of many natural membrane-associated biomolecules. Furthermore, the membrane-bound glycopolymers were internalized into early endosomes similarly to endogenous membrane components and were capable of specific interactions with protein receptors. This system provides a platform to study cell-surface phenomena with a degree of chemical control that cannot be achieved using conventional biological tools.


Attitudes of U.S. and Czech/Slovak mental health and health professionals toward five types of sexually explicit materials.
Mental health and health professionals' attitudes toward sexually explicit materials in the U.S. and Czech/Slovak Republics were investigated. An instrument measuring attitudes toward educational, soft-core, hard-core, violence, and bizarre/paraphiliac sexually explicit materials was administered to sexologists, psychologist/counselors, and medical professionals. These professionals were attending conferences in the U.S. and the Czech/Slovak Republics between November 1992 and September 1993. Mental health and health professionals had the most favorable attitudes toward educational sexually explicit materials followed by soft-core and hard-core materials, respectively. They had unfavorable attitudes toward violent and bizarre/paraphiliac sexually explicit materials, with particularly negative attitudes toward violent materials. Analysis of covariance showed that strength of religious conviction was a significant covariate; thus professionals with stronger religious conviction had more negative attitudes toward all five types of sexually explicit materials. When controlling for strength of religious conviction: (i) sexologists had more positive attitudes toward most types of sexually explicit materials; (ii) Czech professionals generally had more positive attitudes toward such materials than their U.S. counterparts; and (iii) there were few differences between female and male professionals in their reported attitudes. While previous literature has reported gender differences in attitudes toward sexually explicit materials, findings from this study suggest that this effect may be due to differences in religiosity among women and men, namely, that women tend to be more religious.


Aortic compliance in human hypertension.
Aortic compliance in normotensive and hypertensive Chinese subjects undergoing diagnostic cardiac catheterization was compared by using a newly described method that allows for determination of the pressure dependence of compliance if one assumes a value for the exponential coefficient of the pressure-volume relation of the large arteries. Under baseline conditions in the normotensive and hypertensive groups at mean aortic pressures of 96.3 and 128.6 mm Hg, aortic compliance averaged 1.47 and 0.80 ml/mm Hg, respectively. Compliance in the hypertensive group at a diastolic pressure of 99.4 mm Hg (which was nearly equal to the mean normotensive pressure) was 1.072 ml/mm Hg--still significantly lower than in the normotensive group. During nitroprusside infusion, however, the compliances in the hypertensive group increased to levels equal to or greater those in the normotensive group. Thus, these data confirm that aortic compliance is lower in hypertensive than in normotensive humans. They further demonstrate that the lower compliance cannot be attributed entirely to the elevated blood pressure, suggesting that excess smooth muscle tone may be partly responsible.


Synaptic lability after experience-dependent plasticity is not mediated by calcium-permeable AMPARs.
Activity- or experience-dependent plasticity has been associated with the trafficking of calcium-permeable α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (CP-AMPARs) in a number of experimental systems. In some cases it has been shown that CP-AMPARs are only transiently present and can be removed in an activity-dependent manner. Here we test the hypothesis that the presence of CP-AMPARs confers instability onto recently potentiated synapses. Previously we have shown that altered sensory input (single-whisker experience; SWE) strengthens layer 4-2/3 excitatory synapses in mouse primary somatosensory cortex, in part by the trafficking of CP-AMPARs. Both in vivo and in vitro, this potentiation is labile, and can be depressed by N-Methyl-D-aspartate receptor (NMDAR)-activation. In the present study, the role of CP-AMPARs in conferring this synaptic instability after in vivo potentiation was evaluated. We develop an assay to depress the strength of individual layer 4-2/3 excitatory synapses after SWE, using a strontium (Sr(++))-replaced artificial cerebrospinal fluid (ACSF) solution (Sr-depression). This method allows disambiguation of changes in quantal amplitude (a post-synaptic measure) from changes in event frequency (typically a presynaptic phenomenon). Presynaptic stimulation paired with post-synaptic depolarization in Sr(++) lead to a rapid and significant reduction in EPSC amplitude with no change in event frequency. Sr-depression at recently potentiated synapses required NMDARs, but could still occur when CP-AMPARs were not present. As a further dissociation between the presence of CP-AMPARs and Sr-depression, CP-AMPARs could be detected in some cells from control, whisker-intact animals, although Sr-depression was never observed. Taken together, our findings suggest that CP-AMPARs are neither sufficient nor necessary for synaptic depression after in vivo plasticity in somatosensory cortex. This article is part of a Special Issue entitled "Calcium permeable AMPARs in synaptic plasticity and disease."


A disease- and phosphorylation-related nonmechanical function for keratin 8.
Keratin 8 (K8) variants predispose to human liver injury via poorly understood mechanisms. We generated transgenic mice that overexpress the human disease-associated K8 Gly61-to-Cys (G61C) variant and showed that G61C predisposes to liver injury and apoptosis and dramatically inhibits K8 phosphorylation at serine 73 (S73) via stress-activated kinases. This led us to generate mice that overexpress K8 S73-to-Ala (S73A), which mimicked the susceptibility of K8 G61C mice to injury, thereby providing a molecular link between K8 phosphorylation and disease-associated mutation. Upon apoptotic stimulation, G61C and S73A hepatocytes have persistent and increased nonkeratin proapoptotic substrate phosphorylation by stress-activated kinases, compared with wild-type hepatocytes, in association with an inability to phosphorylate K8 S73. Our findings provide the first direct link between patient-related human keratin variants and liver disease predisposition. The highly abundant cytoskeletal protein K8, and possibly other keratins with the conserved S73-containing phosphoepitope, can protect tissue from injury by serving as a phosphate "sponge" for stress-activated kinases and thereby provide a novel nonmechanical function for intermediate filament proteins.


Antagonism of microRNA function in zebrafish embryos by using locked nucleic acid enzymes (LNAzymes).
MicroRNAs (miRNAs) have crucial functions in many cellular processes, such as differentiation, proliferation and apoptosis; aberrant expression of miRNAs has been linked to human diseases, including cancer. Tools that allow specific and efficient knockdown of miRNAs would be of immense importance for exploring miRNA function. Zebrafish serves as an excellent vertebrate model system to understand the functions of miRNAs involved in a variety of biological processes. We designed and employed a strategy based on locked nucleic acid enzymes (LNAzymes) for in vivo knockdown of miRNA in zebrafish embryos. We demonstrate that LNAzyme can efficiently knockdown miRNAs with minimal toxicity to the zebrafish embryos.


Phenol-benzene complexation dynamics: quantum chemistry calculation, molecular dynamics simulations, and two dimensional IR spectroscopy.
Molecular dynamics (MD) simulations and quantum mechanical electronic structure calculations are used to investigate the nature and dynamics of the phenol-benzene complex in the mixed solvent, benzene/CCl4. Under thermal equilibrium conditions, the complexes are continuously dissociating and forming. The MD simulations are used to calculate the experimental observables related to the phenol hydroxyl stretching mode, i.e., the two dimensional infrared vibrational echo spectrum as a function of time, which directly displays the formation and dissociation of the complex through the growth of off-diagonal peaks, and the linear absorption spectrum, which displays two hydroxyl stretch peaks, one for the complex and one for the free phenol. The results of the simulations are compared to previously reported experimental data and are found to be in quite reasonable agreement. The electronic structure calculations show that the complex is T shaped. The classical potential used for the phenol-benzene interaction in the MD simulations is in good accord with the highest level of the electronic structure calculations. A variety of other features is extracted from the simulations including the relationship between the structure and the projection of the electric field on the hydroxyl group. The fluctuating electric field is used to determine the hydroxyl stretch frequency-frequency correlation function (FFCF). The simulations are also used to examine the number distribution of benzene and CCl4 molecules in the first solvent shell around the phenol. It is found that the distribution is not that of the solvent mole fraction of benzene. There are substantial probabilities of finding a phenol in either a pure benzene environment or a pure CCl4 environment. A conjecture is made that relates the FFCF to the local number of benzene molecules in phenol's first solvent shell.


Ingestion of casein and whey proteins result in muscle anabolism after resistance exercise.
Determination of the anabolic response to exercise and nutrition is important for individuals who may benefit from increased muscle mass. Intake of free amino acids after resistance exercise stimulates net muscle protein synthesis. The response of muscle protein balance to intact protein ingestion after exercise has not been studied. This study was designed to examine the acute response of muscle protein balance to ingestion of two different intact proteins after resistance exercise. Healthy volunteers were randomly assigned to one of three groups. Each group consumed one of three drinks: placebo (PL; N = 7), 20 g of casein (CS; N = 7), or whey proteins (WH; N = 9). Volunteers consumed the drink 1 h after the conclusion of a leg extension exercise bout. Leucine and phenylalanine concentrations were measured in femoral arteriovenous samples to determine balance across the leg. Arterial amino acid concentrations were elevated by protein ingestion, but the pattern of appearance was different for CS and WH. Net amino acid balance switched from negative to positive after ingestion of both proteins. Peak leucine net balance over time was greater for WH (347 +/- 50 nmol.min(-1).100 mL(-1) leg) than CS (133 +/- 45 nmol.min(-1).100 mL(-1) leg), but peak phenylalanine balance was similar for CS and WH. Ingestion of both CS and WH stimulated a significantly larger net phenylalanine uptake after resistance exercise, compared with the PL (PL -5 +/- 15 mg, CS 84 +/- 10 mg, WH 62 +/- 18 mg). Amino acid uptake relative to amount ingested was similar for both CS and WH (approximately 10-15%). Acute ingestion of both WH and CS after exercise resulted in similar increases in muscle protein net balance, resulting in net muscle protein synthesis despite different patterns of blood amino acid responses.


Intracerebral gangliogliomas in patients with partial complex seizures: CT and MR imaging findings.
The clinical and radiologic findings in 19 patients with partial complex seizures and surgically proved intracerebral gangliogliomas were reviewed to characterize the radiologic features of these lesions. The CT and MR findings were not specific. On CT the gangliogliomas can be hypodense with no enhancement and they often have calcifications. On MR these tumors have a wide variety of signals. In five of our cases the tumor had a high-intensity signal with a cystlike component on proton density- and T2-weighted images. In five cases the lesion had an inhomogeneously intense signal on proton density-weighted images and high signal intensity on T2-weighted images. The tumor had high-intensity signal on both proton density- and T2-weighted images in four patients. Finally, in two cases the MR findings were normal. We recommend MR as the examination of choice for patients with partial complex seizures because it allows an artifact-free evaluation of the temporal region. However, CT should also be performed in order to recognize calcifications that may be missed on the MR examination.


The hypothalamic leptin receptor in humans: identification of incidental sequence polymorphisms and absence of the db/db mouse and fa/fa rat mutations.
Leptin-receptor gene expression in hypothalamic tissue from lean and obese humans was examined. The full-length leptin receptor, that is believed to transmit the leptin signal, is expressed in human hypothalamus. There was no difference in the amount of leptin-receptor mRNA In seven lean (BMI 23.3 +/- 0.9 kg/m2) and eight obese (BMI 36.9 +/- 1.5) subjects as determined by reverse transcription-polymerase chain reaction. A sequence polymorphism (A-->G) was detected at position 668 of the leptin receptor cDNA. This second base substitution changed a glutamine to an arginine at position 223 of the leptin receptor protein. Of 15 subjects analyzed, 11 were heterozygous for this base change and 3 were homozygous. The occurrence [correction of occurance] of the polymorphic allele(s) did not correlate with BMI in the population studied. The mutation responsible for the defect in the leptin receptor in db/db mice was not detected in any obese human, nor was the fa/fa rat mutation. These results provide evidence that the leptin resistance observed in obese humans is not due to a defect in the leptin receptor.


Impact of posttranslational modifications of engineered cysteines on the substituted cysteine accessibility method: evidence for glutathionylation.
The substituted cysteine accessibility method (SCAM) is widely used to study the structure and function of channels, receptors and transporters. In its usual application, a cysteine residue is introduced into a protein which lacks native cysteines following which the accessibility of the residue to the aqueous compartment is assessed. Implicit, and generally assumed, is that if the cysteine-substituted residue is not available to react with sulfhydryl reagents it is not exposed to the extracellular compartment or within the aqueous translocation pathway. We demonstrate here, in a Hela-derived cell line, that some cysteine-substituted residues of the proton-coupled folate transporter (PCFT, SLC46A1) that are inaccessible to 2-((biotinoyl)amino)ethyl methanethiosulfonate are glutathionylated by biotinylated glutathione ethyl ester in the absence of an oxidizing agent. Intramolecular disulfide formation involving cysteine-substituted residues was also identified in some instances. These posttranslational modifications limit the accessibility of the cysteine residues to sulfhydryl-reactive reagents and can have a profound impact on the interpretation of SCAM but may not alter function. When a posttranslationally modified residue is used as a reference extracellular control, the high level of exposure required for detection on Western blot results in erroneous detection of otherwise inaccessible intracellular cysteine-substituted residues. The data indicate that in the application of SCAM, when a cysteine-substituted residue does not appear to be accessible to sulfhydryl-reactive reagents, the possibility of a posttranslational modification should be excluded. The data explain the discrepancies in the assessment, and confirm the localization, of the first intracellular loop of PCFT.


The impact of Ethiopia's pilot community based health insurance scheme on healthcare utilization and cost of care.
In June 2011, the Government of Ethiopia introduced a pilot Community Based Health Insurance (CBHI) scheme in rural parts of the country. Based on a fixed effects analysis of household panel data, this paper assesses the impact of the scheme on utilization of modern healthcare and the cost of accessing healthcare. It adds to the relatively small body of work that provides a rigorous evaluation of CBHI schemes. We find that in the case of public health facilities, enrolment leads to a 30-41% increase in utilization of outpatient care, a 45-64% increase in the frequency of visits and at least a 56% decline in the cost per visit. The impact on utilization and costs combined with a high uptake rate of almost 50% within two years of scheme establishment underlines the relative success of the Ethiopian scheme. While there are several reasons for this success, a comparative analysis of the design and execution of the Ethiopia CBHI with the existing body of work yields two distinct features. First, the Ethiopian scheme is embedded within existing government administrative structures and to signal government commitment, scheme performance and uptake is used as a yardstick to measure the success of the administration. Second, an existing social protection scheme was used to spread information, raise scheme awareness and encourage uptake of health insurance. The alignment of the interests of administrators with scheme performance and interlinking of social protection schemes are innovative design features that are worth considering as developing countries strive to enhance access to health care through voluntary insurance schemes.


Caffeine decreases vitamin D receptor protein expression and 1,25(OH)2D3 stimulated alkaline phosphatase activity in human osteoblast cells.
Of the various risk factors contributing to osteoporosis, dietary/lifestyle factors are important. In a clinical study we reported that women with caffeine intakes >300 mg/day had higher bone loss and women with vitamin D receptor (VDR) variant, tt were at a greater risk for this deleterious effect of caffeine. However, the mechanism of how caffeine effects bone metabolism is not clear. 1,25-Dihydroxy vitamin D(3) (1,25(OH)(2)D(3)) plays a critical role in regulating bone metabolism. The receptor for 1,25(OH)(2)D(3), VDR has been demonstrated in osteoblast cells and it belongs to the superfamily of nuclear hormone receptors. To understand the molecular mechanism of the role of caffeine in relation to bone, we tested the effect of caffeine on VDR expression and 1,25(OH)(2)D(3) mediated actions in bone. We therefore examined the effect of different doses of caffeine (0.2, 0.5, 1.0 and 10mM) on 1,25(OH)(2)D(3) induced VDR protein expression in human osteoblast cells. We also tested the effect of different doses of caffeine on 1,25(OH)(2)D(3) induced alkaline phosphatase (ALP) activity, a widely used marker of osteoblastic activity. Caffeine dose dependently decreased the 1,25(OH)(2)D(3) induced VDR expression and at concentrations of 1 and 10mM, VDR expression was decreased by about 50-70%, respectively. In addition, the 1,25(OH)(2)D(3) induced alkaline phosphatase activity was also reduced at similar doses thus affecting the osteoblastic function. The basal ALP activity was not affected with increasing doses of caffeine. Overall, our results suggest that caffeine affects 1,25(OH)(2)D(3) stimulated VDR protein expression and 1,25(OH)(2)D(3) mediated actions in human osteoblast cells.


Effect of telomere length on survival in patients with idiopathic pulmonary fibrosis: an observational cohort study with independent validation.
Short telomere lengths are found in a subset of patients with idiopathic pulmonary fibrosis, but their clinical significance is unknown. Our aim was to investigate whether patients with various blood leucocyte telomere lengths had different overall survival. In this observational cohort study, we enrolled patients with interstitial lung disease from Dallas, TX (primary cohort), and from Chicago, IL, and San Francisco, CA (replication cohorts). We obtained genomic DNA samples from unrelated healthy controls in Dallas, TX, and spouses of patients were also enrolled as an independent control group. Telomere lengths were measured in genomic DNA samples isolated from peripheral blood obtained at the time of the initial enrolment assessment. The primary endpoint was transplant-free survival (ie, time to death or lung transplantation) in the Dallas cohort. Findings were validated in the two independent idiopathic pulmonary fibrosis cohorts (Chicago and San Francisco). 370 patients were enrolled into the Dallas cohort between June 17, 2003, and Aug 25, 2011. The 149 patients with idiopathic pulmonary fibrosis had shorter telomere lengths than did the 195 healthy controls (mean age-adjusted log-transformed ratio of telomere to single copy gene was -0.16 [SD 0.23] vs 0.00 [0.18]; p<0.0001); however, telomere lengths of the Dallas patients with idiopathic pulmonary fibrosis (1.33 [SD 0.25]) were similar to the 221 patients with other interstitial lung disease diagnoses (1.46 [0.24]) after adjusting for age, sex, and ethnicity (p=0.47). Telomere length was independently associated with transplant-free survival time for patients with idiopathic pulmonary fibrosis (HR 0.22 [95% CI 0.08-0.63]; p=0.0048), but not for patients with interstitial lung disease diagnoses other than idiopathic pulmonary fibrosis (HR 0.73 [0.16-3.41]; p=0.69). The association between telomere length and survival in patients with idiopathic pulmonary fibrosis was independent of age, sex, forced vital capacity, or diffusing capacity of carbon monoxide, and was replicated in the two independent idiopathic pulmonary fibrosis replication cohorts (Chicago cohort, HR 0.11 [0.03-0.39], p=0.00066; San Francisco cohort, HR 0.25 [0.07-0.87], p=0.029). Shorter leucocyte telomere lengths are associated with worse survival in idiopathic pulmonary fibrosis. Additional studies will be needed to establish clinically relevant thresholds for telomere length and how this biomarker might affect risk stratification of patients with idiopathic pulmonary fibrosis. US National Heart, Lung, and Blood Institute, National Center for Advancing Translational Sciences, Harroun Family Foundation, and Nina Ireland Lung Disease Program.


Amblyomma americanum (Acari: Ixodidae) Ticks Are Not Vectors of the Lyme Disease Agent, Borrelia burgdorferi (Spirocheatales: Spirochaetaceae): A Review of the Evidence.
In the early 1980s, Ixodes spp. ticks were implicated as the key North American vectors of Borrelia burgdorferi (Johnson, Schmid, Hyde, Steigerwalt and Brenner) (Spirocheatales: Spirochaetaceae), the etiological agent of Lyme disease. Concurrently, other human-biting tick species were investigated as potential B. burgdorferi vectors. Rashes thought to be erythema migrans were observed in patients bitten by Amblyomma americanum (L.) (Acari: Ixodidae) ticks, and spirochetes were visualized in a small percentage of A. americanum using fluorescent antibody staining methods, sparking interest in this species as a candidate vector of B. burgdorferi. Using molecular methods, the spirochetes were subsequently described as Borrelia lonestari sp. nov. (Spirocheatales: Spirochaetaceae), a transovarially transmitted relapsing fever Borrelia of uncertain clinical significance. In total, 54 surveys from more than 35 research groups, involving more than 52,000 ticks, have revealed a low prevalence of B. lonestari, and scarce B. burgdorferi, in A. americanum. In Lyme disease-endemic areas, A. americanum commonly feeds on B. burgdorferi-infected hosts; the extremely low prevalence of B. burgdorferi in this tick results from a saliva barrier to acquiring infection from infected hosts. At least nine transmission experiments involving B. burgdorferi in A. americanum have failed to demonstrate vector competency. Advancements in molecular analysis strongly suggest that initial reports of B. burgdorferi in A. americanum across many states were misidentified B. lonestari, or DNA contamination, yet the early reports continue to be cited without regard to the later clarifying studies. In this article, the surveillance and vector competency studies of B. burgdorferi in A. americanum are reviewed, and we conclude that A. americanum is not a vector of B. burgdorferi.


Hepatocellular carcinoma is accelerated by NASH involving M2 macrophage polarization mediated by hif-1αinduced IL-10.
Obesity-related inflammation promotes cancer development. Tissue resident macrophages affect tumor progression and the tumor micro-environment favors polarization into alternatively activated macrophages (M2) that facilitate tumor invasiveness. Here, we dissected the role of western diet-induced NASH in inducing macrophage polarization in a carcinogen initiated model of hepatocellular carcinoma (HCC). Adult C57BL/6 male mice received diethyl nitrosamine (DEN) followed by 24 weeks of high fat-high cholesterol-high sugar diet (HF-HC-HSD). We assessed liver MRI and histology, serum ALT, AFP, liver triglycerides, and cytokines. Macrophage polarization was determined by IL-12/TNFα (M1) and CD163/CD206 (M2) expression using flow cytometry. Role of hif-1α-induced IL-10 was dissected in hepatocyte specific hif-1αKO and hif-1αdPA (over-expression) mice. The western diet-induced features of NASH and accelerated HCC development after carcinogen exposure. Liver fibrosis and serum AFP were significantly increased in DEN + HF-HC-HSD mice compared to controls. Western diet resulted in macrophage (F4/80+CD11b+) infiltration to liver and DEN + HF-HC-HSD mice showed preferential increase in M2 macrophages. Isolated hepatocytes from western diet fed mice showed significant upregulation of the hypoxia-inducible transcription factor, hif-1α, and livers from hif-1α over-expressing mice had increased proportion of M2 macrophages. Primary hepatocytes from wild-type mice treated with DEN and palmitic acid in vitro showed activation of hif-1α and induction of IL-10, a M2 polarizing cytokine. IL-10 neutralization in hepatocyte-derived culture supernatant prevented M2 macrophage polarization and silencing hif-1α in macrophages blocked their M2 polarization. Therefore, our data demonstrate that NASH accelerates HCC progression via upregulation of hif-1α mediated IL-10 polarizing M2 macrophages.


Value of serum glucose-6-phosphate isomerase in patients with rheumatoid arthritis and correlation with disease activity: A case-control study.
Rheumatoid arthritis (RA) is a common multisystemic autoimmune disease with peripheral joint involvement. Many autoantibodies have been introduced in the course of RA; some of them have diagnostic and prognostic value. In this study, our aim is to determine the value of serum glucose-6-phosphate isomerase (G6PI) antigen (Ag) as a diagnostic and prognostic marker in RA. Eighty-seven known cases of RA who referred to an outpatient clinic of Shiraz University of Medical Sciences and 76 healthy controls were selected. Serum G6PI Ag was measured using sandwich enzyme-linked immunosorbent assay method, and the enzyme level was compared in the patient and control group, we also compared the enzyme level of patient group with disease activity, disease duration, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), rheumatoid factor (RF), and anti-cyclic citrullinated peptide (anti-CCP) antibody (Ab). The data were analyzed using SPSS V 16 software. Positivity for G6PI was detected in 34.5% (30/87) of RA patients and 9.2% (7/76) of control group (P < 0.001). There was no significant correlation between enzyme level and disease activity, disease duration, ESR, CRP, RF, and anti-CCP Ab. Overall, in our study, although there was a significant difference in serum G6PI Ag between patient and control group, no significant correlation was detected between serum G6PI level and disease activity score, ESR, CRP, and anti-CCP Ab, but relative correlation with ESR and disease duration could be challenging. G6PI Ag could be introduced as a diagnostic marker in RA, but its role as a prognostic marker is controversial.


First identification and characterization of Borrobol-type tephra in the Greenland ice cores: new deposits and improved age estimates.
Contiguous sampling of ice spanning key intervals of the deglaciation from the Greenland ice cores of NGRIP, GRIP and NEEM has revealed three new silicic cryptotephra deposits that are geochemically similar to the well-known Borrobol Tephra (BT). The BT is complex and confounded by the younger closely timed and compositionally similar Penifiler Tephra (PT). Two of the deposits found in the ice are in Greenland Interstadial 1e (GI-1e) and an older deposit is found in Greenland Stadial 2.1 (GS-2.1). Until now, the BT was confined to GI-1-equivalent lacustrine sequences in the British Isles, Sweden and Germany, and our discovery in Greenland ice extends its distribution and geochemical composition. However, the two cryptotephras that fall within GI-1e ice cannot be separated on the basis of geochemistry and are dated to 14358 ± 177 a b2k and 14252 ± 173 a b2k, just 106 ± 3 years apart. The older deposit is consistent with BT age estimates derived from Scottish sites, while the younger deposit overlaps with both BT and PT age estimates. We suggest that either the BT in Northern European terrestrial sequences represents an amalgamation of tephra from both of the GI-1e events identified in the ice-cores or that it relates to just one of the ice-core events. A firm correlation cannot be established at present due to their strong geochemical similarities. The older tephra horizon, found within all three ice-cores and dated to 17326 ± 319 a b2k, can be correlated to a known layer within marine sediment cores from the North Iceland Shelf (ca. 17179-16754 cal a BP). Despite showing similarities to the BT, this deposit can be distinguished on the basis of lower CaO and TiO2 and is a valuable new tie-point that could eventually be used in high-resolution marine records to compare the climate signals from the ocean and atmosphere.


Lantana camara L. (Verbenaceae) invasion along streams in a heterogeneous landscape.
Streams are periodically disturbed due to flooding, act as edges between habitats and also facilitate the dispersal of propagules, thus being potentially more vulnerable to invasions than adjoining regions. We used a landscape-wide transect-based sampling strategy and a mixed effects modelling approach to understand the effects of distance from stream, a rainfall gradient, light availability and fire history on the distribution of the invasive shrub Lantana camara L.(lantana) in the tropical dry forests of Mudumalai in southern India. The area occupied by lantana thickets and lantana stem abundance were both found to be highest closest to streams across this landscape with a rainfall gradient. There was no advantage in terms of increased abundance or area occupied by lantana when it grew closer to streams in drier areas as compared to moister areas. On an average, the area covered by lantana increased with increasing annual rainfall. Areas that experienced greater number of fires during 1989-2010 had lower lantana stem abundance irrespective of distance from streams. In this landscape, total light availability did not affect lantana abundance. Understanding the spatially variable environmental factors in a heterogeneous landscape influencing the distribution of lantana would aid in making informed management decisions at this scale.


Rotavirus vaccination of very low birth weight infants at discharge from the NICU.
Preterm infants are at increased risk of severe rotavirus gastroenteritis. Although immunization with rotavirus vaccine is safe and effective, age restrictions limit the number of infants eligible for vaccination at discharge from the NICU. The objectives of this study were to assess the implementation of the rotavirus vaccine program in our NICU, recognize missed opportunities for vaccination, and document how often very low birth weight (VLBW; birth weight ≤1500 g) and extremely low birth weight (ELBW; birth weight <1000 g) infants were eligible to receive rotavirus vaccine at the time of NICU discharge. This study reports on a prospective, observational cohort of all VLBW infants who were discharged from the NICU at Parkland Memorial Hospital from May 2008 to April 2010. Medical records were reviewed and data collected regarding the number of infants who were eligible for and received rotavirus vaccination at discharge. A total of 63% (135 of 213) of VLBW infants did not receive rotavirus vaccine. The reasons for not providing vaccine included the following: <42 days of age at discharge (56 of 213; 26%), >84 or 104 days of age at discharge (48 of 213; 23%), or missed (35 of 213; 16%). The majority (75%) who were too old for vaccination at the time of discharge were ELBW. The current age restrictions for rotavirus immunization resulted in more than half of ELBW infants being ineligible for vaccination at the time of discharge from the NICU. Alternative strategies for rotavirus immunization in this population are needed.


The effective concentration of unbound ink anchors at the molecular printboard.
Self-assembled monolayers terminating in beta-cyclodextrin cavities can be used to bind ink molecules and so provide a molecular printboard for nanopatterning applications. Multivalent or multisite binding strengthens the attachment of large inks and provides more robust patterns. In the present work we use computer simulations to probe the behavior of functionalized dendrimer inks at the printboard. We performed a series of long 10 ns fully atomistic molecular dynamics (MD) simulations to measure the effective local concentration of unbound ink anchor groups at the printboard for a variety of binding modes and also for the partial unbinding prerequisite for ink diffusion on the printboard. These simulations allow us to describe the conformational space occupied by partially bound inks and estimate the likelihood of an additional binding interaction. Furthermore, by simulating the shift from a divalent to monovalent binding mode we show that the released anchor quickly moves to the periphery of the dendrimer binding hemisphere but then reapproaches the printboard and remains in the vicinity of alternative binding sites. Secondary electrostatic interactions between the protonated dendrimer core and hydroxyl groups at the entrance to the beta-cyclodextrin cavities give "flattened" dendrimer binding orientations and may aid dendrimer diffusion on the printboard, allowing the dendrimer to "walk" along the printboard by switching between different partially bound states and minimizing complete unbinding to bulk solution, crucial for the application of the printboard in, for example, medical diagnostics.


Association between cancer, CHA2DS2VASc risk and In-hospital ischemic stroke in patients hospitalized for atrial fibrillation.
Atrial fibrillation (AF) is commonly encountered in cancer patients. We investigated the CHA2DS2VASc score, and its association with in-hospital ischemic stroke in patients with cancer who were hospitalized for AF. Using the United States National Inpatient Sample, all hospitalizations with principal diagnosis of AF between October 2015 to December 2018 were stratified by cancer diagnosis, type and CHA2DS2VASc risk categories (low risk; low-moderate risk; moderate-high risk). In-hospital ischemic stroke and its association with the CHA2DS2VASc risk score was assessed across the groups using hierarchical multivariable logistic regression with adjusted odds ratios (aOR) and 95% confidence intervals (95% CI). Discrimination of CHA2DS2VASc score for in-hospital ischemic stroke was evaluated with Receiver Operating Characteristic and Area Under the Curve (AUC). Among 1 341 870 included hospitalizations, 71 965 (5.4%) had comorbid cancer. Cancer patients had a higher proportion of moderate-high CHA2DS2VASc risk compared to their non-cancer counterparts (86.5% vs. 82.3%, p < 0.001). Compared to their low CHA2DS2VASc risk counterparts, cancer patients in low-moderate and moderate-high risk scores had similar odds of developing stroke (aOR 1.28 95% CI 0.22-7.63 and aOR 1.78 95% CI 0.41-7.66, respectively). The CHA2DS2VASc risk score had poor discrimination for ischemic stroke in the cancer group (AUC 0.538 95% CI 0.477-0.598). Cancer patients with AF have high CHA2DS2VASc risk. Discrimination of CHA2DS2VASc for ischemic stroke is lower in cancer than non-cancer patients, and CHA2DS2VASc may not be adequate in determining ischemic risk in cancer population.


Intrinsic pigment cell stimulating activity in the skin of the leopard frog, Rana pipiens.
Consistent with the concept that specific pigment patterns of amphibians might result from the highly localized distribution of stimulators and inhibitors of pigment cell expression in the skin, the spot pattern of the leopard frog, Rana pipiens, was examined through the use of the Xenopus neural tube explant assay system (Fukuzawa and Ide, 1988). Media conditioned with pieces of skin from dorsal black spotted areas promoted melanization of neural crest cells at a significantly higher level than did media conditioned with dorsal interspot skin in the absence of extra tyrosine. All conditioned media contained exceedingly low concentrations of tyrosine. With the addition of supplemental tyrosine, the melanization capacity of conditioned media from the interspot areas was elevated to that of the spotted skin. Control media conditioned with ventral frog skin inhibited melanization, as usual, because of the presumed presence of melanization inhibiting factor (MIF). It is considered that dorsal skin contains a melanization stimulating factor (MSF) which is present in significantly higher levels in spotted skin than in interspot areas and that expression of the particular pigmentary pattern of this leopard frog is regulated by the relative distribution of MIF, MSF, and possibly other intrinsic substances present in the skin.


North/South Differences Among Italian Emerging Adults Regarding Criteria Deemed Important for Adulthood and Life Satisfaction.
The main goal of this study was to compare Northern and Southern Italian emerging adult university students, regarding the importance attributed to criteria for adulthood and the levels of life and education satisfaction. Self-report questionnaires were filled by 475 Northern and Southern Italian University students (Age M = 22.91, 76% females, n = 359). Multivariate analysis of variance revealed that Southern emerging adults were more likely to place importance on family capacities, norm compliance, interdependence and role transitions as criteria for achieving adulthood than Northern emerging adults. Regarding gender differences, females were more likely to believe in the importance of norm compliance than males, while males were more likely to espouse the importance of legal transitions. Finally, emerging adults from the North reported higher levels of life satisfaction than their Southern counterparts. We interpreted these findings in light of socio-economical and gender socialization differences among Northern and Southern Italian emerging adults.


Herpes simplex virus and HIV-1: deciphering viral synergy.
Recent proof-of-concept randomised controlled trials have shown a causal relation between herpes simplex virus (HSV) type 2 infection and HIV-1 replication in co-infected individuals. We explore the mechanisms that may operate to enhance reciprocal viral replication. Direct interactions could involve HIV-1-related immune deficiency, disruption of mucosal barrier by HSV infection/reactivation, HSV-induced mucosal cell recruitment, transactivation of HIV-1 replication by HSV proteins, and immune modulation by HSV decoys. Indirect interactions might coexist through disturbances of the vaginal flora during HSV shedding and systemic immune activation. In co-infected individuals, suppressive HSV treatment reduces HIV-1 genital and systemic excretion. This finding is a likely result of efficacious prevention of HSV2 reactivations, and perhaps of other herpesviruses. Strategies to control HSV2 and other herpesviruses deserve urgent attention and should become part of the HIV-1 prevention and care package.


Functional characterization of the SIZ/PIAS-type SUMO E3 ligases, OsSIZ1 and OsSIZ2 in rice.
Sumoylation is a post-translational regulatory process in diverse cellular processes in eukaryotes, involving conjugation/deconjugation of small ubiquitin-like modifier (SUMO) proteins to other proteins thus modifying their function. The PIAS [protein inhibitor of activated signal transducers and activators of transcription (STAT)] and SAP (scaffold attachment factor A/B/acinus/PIAS)/MIZ (SIZ) proteins exhibit SUMO E3-ligase activity that facilitates the conjugation of SUMO proteins to target substrates. Here, we report the isolation and molecular characterization of Oryza sativa SIZ1 (OsSIZ1) and SIZ2 (OsSIZ2), rice homologs of Arabidopsis SIZ1. The rice SIZ proteins are localized to the nucleus and showed sumoylation activities in a tobacco system. Our analysis showed increased amounts of SUMO conjugates associated with environmental stresses such as high and low temperature, NaCl and abscisic acid (ABA) in rice plants. The expression of OsSIZ1 and OsSIZ2 in siz1-2 Arabidopsis plants partially complemented the morphological mutant phenotype and enhanced levels of SUMO conjugates under heat shock conditions. In addition, ABA-hypersensitivity of siz1-2 seed germination was partially suppressed by OsSIZ1 and OsSIZ2. The results suggest that rice SIZ1 and SIZ2 are able to functionally complement Arabidopsis SIZ1 in the SUMO conjugation pathway. Their effects on the Arabidopsis mutant suggest a function for these genes related to stress responses and stress adaptation.


A plasmid-based multigene expression system for mammalian cells.
The introduction of heterologous genetic information, particularly of multiple genes, into mammalian cells is a key technology in contemporary experimental biological research. The coexpression of fluorescently tagged sensors is required to simultaneously analyse multiple parameters in living cells and the coexpression of several proteins is necessary to manipulate cell fate in stem cell biology. Current technologies for multigene expression in mammalian cells are inefficient, inflexible and time-consuming. In this paper we describe MultiLabel, a novel and highly efficient modular plasmid-based eukaryotic expression system. Independent expression vectors are assembled by a Cre/LoxP reaction into a plasmid with multiple expression cassettes. MultiLabel enables rapid construction of multigene expression vectors for the single-step creation of transiently or stably transfected mammalian cells.


Diversity of carbapenem resistance mechanisms in Acinetobacter baumannii from a Taiwan hospital: spread of plasmid-borne OXA-72 carbapenemase.
We investigated the molecular epidemiology of carbapenem-resistant Acinetobacter baumannii from a Taiwanese hospital and determined the mechanisms responsible for resistance. Ninety-two consecutive meropenem-resistant A. baumannii isolates collected between January 2005 and June 2007 were screened for genes encoding OXA carbapenemases, metallo-beta-lactamases and for the carO gene encoding an outer membrane protein. PFGE was used to define clonal relatedness. PCR mapping was used to examine the linkage of insertion sequences and bla(OXA) genes. Southern hybridization of plasmid extracts and I-CeuI-restricted chromosomal DNA was used to locate bla(OXA-24-like) genes. Sequences of selected bla(OXA-24-like) and carO genes were determined and loss of CarO expression was confirmed by SDS-PAGE. Most (70/92, 76%) isolates belonged to one of three PFGE pulsotypes, indicating clonal spread. Fifty-nine isolates, including the majority of those of pulsotypes I and III, produced OXA-72 carbapenemase. The bla(OXA-72) gene was located on a 54 kb plasmid in selected isolates. Thirty-three (36%) isolates, including all 16 of pulsotype II, had ISAba1 preceding the bla(OXA-51-like) gene, promoting its expression. In addition to OXA-72 carbapenemase, two pulsotype I and three pulsotype III isolates did not produce CarO protein as the carO gene was disrupted by insertion of an ISAba1 element. Two isolates of a minor pulsotype had a bla(OXA-58-like) gene and a single PFGE-unique isolate had a bla(OXA-23-like) gene. Although diverse mechanisms were identified, production of OXA-72 carbapenemase was the most common mechanism of carbapenem resistance in A. baumannii from this Taiwanese hospital. The plasmidic location of the gene had facilitated its spread to multiple strains.