The importance of cigarette packaging in a 'dark' market: the 'Silk Cut' experience.
In a growing number of countries tobacco companies are severely restricted in how they can legally market their products. In these 'dark' markets the role of packaging as a promotional and communications tool becomes more pronounced. How packaging is used for the most expensive cigarette brands in dark markets has received limited attention however, even though these 'premium' cigarette brands significantly impact upon the profitability of tobacco companies. We outline, using retail trade press journals, how packaging was used for premium brand 'Silk Cut' in the UK from 2004 to 2011, following a comprehensive ban on tobacco advertising, promotions and sponsorship. From 2004 to 2008 packaging was used to help launch two new variants and during this period Silk Cut market share of the premium sector grew by 1.1%. Overall share of the cigarette market for the Silk Cut house (brand family) fell however due to the continuing decline of the premium sector. From 2008 to 2011 changes to the packaging were much more frequent, including the repeated use of limited-edition designs, and modifications to pack shape, texture, style of opening, cellophane, foil and inner frame. Silk Cut's share of the premium sector grew a further 2.9% from 2008 to 2011, and overall cigarette market share increased. That a premium brand can report any level of growth within such a hostile market, where most advertising, promotion and sponsorship is banned, taxation is among the highest in the world, and in the midst of a recession, is testament to the value of packaging.


Strapping rowers to their sliding seat improves performance during the start of single-scull rowing.
In this study, the effect of strapping rowers to their sliding seat on performance during 75 m on-water starting trials was investigated. Well-trained rowers performed 75 m maximum-effort starts using an instrumented single scull equipped with a redesigned sliding seat system, both under normal conditions and while strapped to the sliding seat. Strapping rowers to their sliding seat resulted in a 0.45 s lead after 75 m, corresponding to an increase in average boat velocity of about 2.5%. Corresponding effect sizes were large. No significant changes were observed in general stroke cycle characteristics. No indications of additional boat heaving and pitching under strapped conditions were found. The increase in boat velocity is estimated to correspond to an increase in average mechanical power output during the start of on-water rowing between 5% and 10%, which is substantial but smaller than the 12% increase found in a previous study on ergometer starting. We conclude that, after a very short period of adaptation to the strapped condition, single-scull starting performance is substantially improved when the rower is strapped to the sliding seat.


Exosomal delivery of berry anthocyanidins for the management of ovarian cancer.
Despite optimal diagnosis and early therapeutic interventions, the prognosis for ovarian cancer patients remains dismal because the efficacy of chemotherapy is limited by the development of resistance and off-site toxicity. Berry bioactives indicate preventive and therapeutic activities against various cancer types. Here, we examined the antiproliferative activity of berry anthocyanidins (Anthos) against drug-sensitive (A2780) and drug-resistant (A2780/CP70, OVCA432 and OVCA433) ovarian cancer cells. These drug-resistant ovarian cancer cell lines overexpress p-glycoproteins (PgP) and show >100-fold resistance to the chemotherapeutic drug cisplatin compared to A2780. We observed a dose-dependent growth inhibition of ovarian cancer cells with the Anthos. Furthermore, the treatment of drug-resistant ovarian cancer (OVCA433) cells with cisplatin in combination with the Anthos (75 μM) resulted in significantly higher cell killing. The cisplatin dose required to achieve this effect was 10 to 15-fold lower than the IC50 of cisplatin alone. However, many plant bioactives including Anthos face the challenge of poor oral bioavailability and stability. Recently, we have developed strategies to overcome these limitations by delivering Anthos via milk-derived exosomes. The exosomal Anthos (ExoAnthos) significantly enhanced the antiproliferative activity against the growth of ovarian cancer cells and inhibited tumor growth more efficiently compared to Anthos alone and a vehicle control. Often patients with cisplatin-resistant tumors retain sensitivity to paclitaxel (PAC). We prepared exosomal formulations of PAC (ExoPAC) for oral delivery as the systemic administration of PAC has severe side effects. ExoPAC delivered orally showed the same therapeutic efficacy as the free PAC delivered intraperitoneally. Finally, we report that the combination of the Anthos and PAC decreased the PgP level in a dose-dependent manner in OVCA432 cells. A significantly enhanced antitumor activity was observed with the combination of ExoPAC and ExoAnthos against A2780 tumor xenografts. Together, our data indicate that the berry Anthos are highly effective against ovarian cancer and that the milk exosomes serve as an excellent nano-carrier to enhance the drug's oral bioavailability for the management of ovarian cancer.


Overcoming a Bad Day: a Qualitative Look into the Dementia Caregiving Experiences of Mexican-Origin Women in East Los Angeles.
The number of Latinos with Alzheimer's disease (AD) is projected to more than double by 2030. Yet, the current literature is lagging on Alzheimer's caregiving among Latinos. This study explores how Mexican origin women experience dementia caregiving, and the coping strategies they use to manage their caregiving situations. Nine women were identified as caregivers of a family member with AD or dementia from a larger study on caregiving. Interviews with them were collected and subsequently analyzed using a modified grounded theory approach to reduce the data to identify thematic content. All but one participant described caregiving as "tiring," "wearing," or "hard." They reported suffering from stress, insomnia, nervousness, migraines, and/or depression because of their caregiving experiences. Participants engaged in various coping strategies to help combat the perceived negative consequences of their caregiving experiences. The most commonly reported strategies were various forms of distraction, and meditation or prayer. The Mexican origin women in this study experience faced two types of interpersonal challenges related to dementia caregiving: changes in the care receiver's personality and behaviors, and physical care needs. They engaged in various coping to strategies to address the difficulties of their situation. This study provides formative research for identifying research questions and topics of examination in the future.


Organ-specific and light-induced expression of plant genes.
Light plays a pivotal role in the development of plants. The photoregulation of plant genes involves recognition of light quality and quantity by phytochrome and other light receptors. Two gene families, rbcS and Cab, which code for abundant proteins active in photosynthesis, the small subunit of ribulose bisphosphate carboxylase and the chlorophyll a/b binding protein, show a 20-to 50-fold increase in transcript abundance in the light. Analyses in calli and transgenic plants of deletions of the rbcS gene and of chimeric constructions has allowed localization of two regions involved in light-induced transcription. One element is confined to a 33-base pair region surrounding the TATA box. In addition, an enhancer-like element contained within a 240-base pair fragment can confer phytochrome-induced transcription and organ specificity on nonregulated promoters.


Residual complaints following lumbar disc surgery: prognostic indicators of outcome.
Physical as well as psychological features might be important prognostic factors for residual complaints following lumbar disc surgery in primary care. No studies have yet investigated both factors simultaneously. The aim of this prospective cohort study was to identify indicators of the short and long-term outcome of residual complaints following lumbar disc surgery. Patients (n=105), aged between 18 and 65 years, were included if they still suffered residual complaints 6 weeks after first-time lumbar disc surgery and had therefore been referred to physiotherapy. All potential indicators were measured at baseline except treatment expectancy, which was measured after two treatment sessions enabling patients to rate treatment expectancy based on their actual perception of the treatment. Dimensions of recovery included perceived recovery, functional status, and pain intensity (back and leg) at the 3-month and 12-month follow-up. It was found that high treatment expectancy was associated with a favorable outcome on perceived recovery and functional status, both at the 3 and the 12-month follow-up. Taking pain medication and a poor functional status at baseline were associated with poor perceived recovery and functional status at both follow-up measurements. Leg pain and back pain at baseline were associated with residual leg and back pain at the 3 and the 12-month follow-up, respectively. The results for perceived recovery and functional status were rather robust. However, for leg pain and back pain, the results were less stable. Apparently, the clinical course to recovery of residual leg pain and residual back pain is not strongly influenced by these indicators.


Erythrocyte membrane enzymes in sickle cell anemia. 2. Acetylcholinesterase and ATPase activities.
The activity level of acetylcholinesterase in the erythrocytes of 32 patients homozygous for sickle cell anemia was determined and compared with that of normal AA controls as well as with that of AS individuals. Acetylcholinesterase activity was markedly higher in erythrocyte membrane from SS individuals than in those from AS individuals or AA controls. Additionally, ATPase activities were also significantly higher in sickle cell erythrocytes as compared to normal cells. These higher values of acetylcholinesterase and ATPase activities in SS erythrocytes may be explained as a consequence of the abnormally high cation levels in sickle cell erythrocytes.


Bullying Involvement in Adolescence: Implications for Sleep, Mental Health, and Academic Outcomes.
Adolescents' involvement in bullying is associated with both sleep and mental health problems, but the nature of this association remains unclear; further, its association with academic outcomes has received little attention. Thus, the aims of the current study were to (a) determine whether involvement in bullying as a victim, bully, or bully-victim was associated with greater sleep and mental health problems and (b) explore the potential mediating effect of sleep and mental health problems on the association between bullying and academic outcomes. A large 2012 population-based study in Hordaland County, Norway, surveyed 10,220 adolescents (16-19 years; 54% girls) about bullying involvement using the revised version of the Olweus Bully/Victim Questionnaire, detailed sleep assessment, and mental health questionnaires. Academic outcomes were obtained from official administrative registries. 1.7% of the adolescents (n = 156) reported being victims of bullying, 1.0% (n = 92) reported being a bully, and 0.5% (n = 50) reported being a bully-victim. All categories of bullying involvement had higher rates of mental health problems compared with adolescents not involved in bullying. Victims reported more symptoms of anxiety and depression, whereas bullies reported higher rates of conduct problems. Adolescents in all bullying categories also reported significantly shorter sleep duration and higher prevalence of insomnia as well as lower grade point average (GPA) compared with adolescents not involved; however, school absence was not associated with bullying involvement. Bullying involvement and GPA showed complete mediation for bullies and bully-victims and partial mediation for victims through sleep duration, conduct problems, and symptoms of depression and attention deficit hyperactivity disorder (ADHD). Bullying is strongly associated with mental health and sleep problems, in addition to lower academic performance. Findings support the importance of addressing bullying involvement during this important developmental period.


Pupillary responses to coloured and contourless displays in total cerebral achromatopsia.
In two patients with total acquired cortical colour blindness and in six control subjects we studied the binocular pupillary response to a variety of sharply defined coloured and grey displays that either had the same mean luminance as the background (isoluminant) or were of greater mean luminance. Despite their complete inability to identify or to discriminate between colours the patients, like the control subjects, showed a pupillary response to the structured coloured displays, even when they were masked by dynamic luminance changes. However, and unlike the control subjects, the patients showed no pupillary response when the coloured displays lacked sharp chromatic borders, as in Gabors or Gaussians. The results indicate that although chromatic processing still occurs in cortical colour blindness its function is solely to give rise to the detection of sharp boundaries which, in their case, can provide the perception of shape but not hue. In accordance with this, the patients could no longer describe the isoluminant borderless figures, which were often totally invisible to them despite their strong chromatic contrast with the background.


Testing social-cognitive mediators for objective estimates of physical activity from the Healthy Eating and Active Living for Diabetes in Primary Care Networks (HEALD-PCN) study.
More evidence from prospective studies is needed to determine 'if' and 'how' social cognitive constructs mediate behaviour change. In a longitudinal study, we aimed to examine potential social cognitive mediators of objectively measured physical activity (PA) behaviour among people with type 2 diabetes (T2D) who participated in a six-month PA intervention. All participants from the proven effective Healthy Eating and Active Living for Diabetes in Primary Care Networks trial were included for this secondary analysis. Change in pedometer-derived daily step counts (baseline to six months) was the outcome of interest. Primary constructs of interest were from Social Cognitive Theory, however constructs from and Theory of Planned Behaviour were also tested in a mediating variable framework using a product-of-coefficients test. The sample (N = 198) had a mean age of 59.5 (SD 8.3) years, haemoglobin A1c 6.8% (SD 1.1), 50% women, BMI 33.6 kg/m(2) (SD 6.5), systolic pressure 125.6 mmHg (SD 16.2) and average daily steps were 5879 (SD 3130). Daily pedometer-determined steps increased for the intervention group compared to usual care control at six-months (1481 [SD 2631] vs. 336 [SD 2712]; adjusted p = .002). There was a significant action theory test effect for 'planning' (A = .21, SE = .10, p = .037), and significant conceptual theory test results for 'subjective norms' (B = 657, SE = 312, p = .037) and 'cons' (B = -664, SE = 270, p = .015). None of the constructs satisfied the criteria for mediation. We were unable to account for the effect of a pedometer-based PA intervention for people with T2D through our examination of mediators. Our findings are inconsistent with some literature concerning PA interventions in diabetes; this may be due to variability in measures used or in study populations.


Detection of Symptomatic Carotid Plaque Using Source Data from MR and CT Angiography: A Correlative Study.
Carotid plaque MRI has been a useful method to characterize vulnerable atherosclerotic plaque elements. Recent investigations have suggested that source images from CT angiography (CTA) and MR angiography (MRA) can identify the simple high-risk features of symptom-producing carotid artery plaque. We studied the correlation and relative diagnostic accuracies of CTA and MRA source images in detecting symptomatic carotid artery plaque. Subjects were eligible if they had carotid stenosis between 50 and 99% and had MRA and CTA exams performed within 10 days of one another. We measured the soft (non-calcified) plaque and hard (calcified) plaque thickness on CTA axial source images and intraplaque high-intensity signal (IHIS) on 3D-time-of-flight MRA source images in subjects. We assessed whether a correlation existed between increasing CTA soft plaque thicknesses and the presence of MRA IHIS using the Student's t-test. We calculated the differences in sensitivity and specificity measures of CTA and MRA source-imaging data with the occurrence of recent ipsilateral stroke or transient ischemic attack (TIA) as the reference standard. We also performed logistic regression analyses to evaluate the predictive strength of plaque showing both IHIS and increased CTA soft plaque thickness in predicting symptomatic disease status. Of 1994 screened patients, 48 arteries met the final inclusion criteria with MRA and CTA performed within 10 days of one another. The mean and median time between CTA and MRA exams were 2.0 days and 1 day, respectively. A total of 34 of 48 stenotic vessels (70.8%) were responsible for giving rise to ipsilateral stroke or TIA. CTA mean soft plaque thickness was significantly greater (4.47 vs. 2.30 mm, p < 0.0001) in patients with MRA-defined IHIS, while CTA hard plaque thickness was significantly greater (2.09 vs. 1.16 mm, p = 0.0134) in patients without MRA evidence of IHIS. CTA soft plaque thickness measurements were more sensitive than MRA IHIS (91.2 vs. 67.6%, p = 0.011) in detecting symptomatic plaque, while differences in specificity were not significantly different (p = 0.1573). In the subset of patients with both IHIS on MRA and plaque thickness >2.4 mm on CTA, the odds ratio of detecting symptomatic plaque, corrected for stenosis severity, was 45.3 (p < 0.0005). Unprocessed source images from CTA and MRA, which are routinely evaluated for clinical studies demonstrate the highly correlated presence of IHIS and increasing soft plaque thickness. In particular, plaque that shows high-risk features on both MRA and CTA are very strongly associated with symptom-producing carotid plaque. With further validation, such techniques are promising practical methods of extracting risk information from routine neck angiographic imaging.


In vitro and in vivo characterization of disulfide bond use in myocilin complex formation.
Myocilin forms large complexes in aqueous humor. Part of this complex formation is due to myocilin-myocilin protein non-covalent interactions within the leucine zipper. However, additional covalent interactions also exist. We investigated the role of these covalent interactions in disulfide bond formation within myocilin. Human aqueous humor was separated by denatured/non-reduced SDS-PAGE followed by Western blot analysis with myocilin specific antibodies. In part two of the study, site-directed mutagenesis was used to selectively mutate one, two, three, four, and all five cysteine residues in the mature myocilin protein expressed in an in vitro system. Products were immunoprecipitated with a hemagglutinin polyclonal antibody following in vitro transcription/translation and analyzed by SDS-PAGE. In part three of the study, glaucoma associated myocilin mutations Arg82Cys and Cys433Arg were created and complex formation analyzed in trabecular meshwork cells. Human aqueous humor showed myocilin in several distinct large complexes in non-reduced SDS-PAGE gels, indicating disulfide bonds occur. Similarly, in vitro expressed myocilin also produced large complexes. Mutation of all five cysteines (within the mature myocilin protein) eliminated this large complex formation. A combination of cysteine to alanine substitutions at amino acids 185, 245, and 433 had the most influence on myocilin complex formation under non-reducing conditions, however individual substitutions at each of the five cysteine amino acids had little influence on myocilin complexes. In trabecular meshwork cells, Arg82Cys was secreted but formed different sized complexes than wild type myocilin. Cys433Arg was not secreted and remained intracellular in a pattern that differed from wild type myocilin and Arg82Cys. Myocilin complexes present in human aqueous humor are in part due to disulfide bond formation between cysteine amino acids. Glaucoma associated mutations that affect the number of cysteine residues may alter covalent interactions.


Calcium ion--the key player in cerebral ischemia.
Role of calcium ion (Ca2+) in the functioning of neurons from their naïve state to mature state is of vital importance. It controls functions such as neuronal functioning, neuronal ATP production, central nervous system migration and many others. Failure in Ca2+ homeostasis mechanisms and the resulting cellular Ca2+ ion load initiates a cascade of reactions involving various cytosolic enzymes and proteins. This total mechanism leads to the neuronal death. The ability of neurons to resist such death mechanisms fails as a result of extensive cell death signaling cascade reactions and later brings brain damage. The role of neuronal endoplasmic reticulum and protein channels like CaVs, TRP channels, and NMDAR as the mediators of cell damage and death has been evaluated in the studies related to cerebral ischemia. Here, we portray Ca2+ ion as one of the role players in neuronal death and cerebral damage following ischemia. The role of Ca2+ in neuronal functioning, its regulatory mechanisms and the failure of homeostatic mechanisms are discussed in detail.


Pre-calculated protein structure alignments at the RCSB PDB website.
With the continuous growth of the RCSB Protein Data Bank (PDB), providing an up-to-date systematic structure comparison of all protein structures poses an ever growing challenge. Here, we present a comparison tool for calculating both 1D protein sequence and 3D protein structure alignments. This tool supports various applications at the RCSB PDB website. First, a structure alignment web service calculates pairwise alignments. Second, a stand-alone application runs alignments locally and visualizes the results. Third, pre-calculated 3D structure comparisons for the whole PDB are provided and updated on a weekly basis. These three applications allow users to discover novel relationships between proteins available either at the RCSB PDB or provided by the user. A web user interface is available at http://www.rcsb.org/pdb/workbench/workbench.do. The source code is available under the LGPL license from http://www.biojava.org. A source bundle, prepared for local execution, is available from http://source.rcsb.org firstname.lastname@example.com; firstname.lastname@example.com.


Fire disaster preparedness and situational analysis in higher learning institutions of Tanzania.
Fire disasters are accompanied with devastating impact affecting both lives and properties. The magnitude of the impacts has been severe in places with low levels of fire disaster preparedness. A study was conducted in Dar es Salaam, Tanzania, to investigate the level of fire disaster preparedness considering the availability and condition of firefighting facilities as well as the knowledge on fire management among the selected 10 higher learning institutions. Information for the buildings was obtained from the interviews with the managers of the buildings and field observations; information on the user's preparedness was obtained from interviews using structured questionnaire conducted with the users of the buildings including the visitors. Results from the studied buildings indicated that 60% of the firefighting facilities were not regularly serviced; 50% stored some hazardous materials; 70% of them had not enough water storage for firefighting purposes; 60% had no identifiable fire assembly points; and 90% of the sessions conducted in the buildings involved more than 100 people in a single venue. Further results indicated that 51% of the respondents were not able to operate the installed firefighting facilities; 80.7% of the respondents had never received any training on firefighting and prevention; 95.6% of the respondents had never participated in any fire drills; and 81.5% of them were not aware of the fire responder's contacts. General situation indicated that higher learning institutions are not well prepared to manage fire outbreaks suggesting that plans to rectify the situation are imperative.


Inhibition of MMP-2-Mediated Mast Cell Invasion by NF-κB Inhibitor DHMEQ in Mast Cells.
Stimulation with antigen and IgE is known to activate NF-κB in mast cells. In the present research, we studied the role of NF-κB on cellular migration in mast cell-like RBL-2H3 cells and bone marrow-derived mast cells (BMMCs) using the NF-κB inhibitor (-)-DHMEQ. A Matrigel invasion chamber was used to evaluate cell migration. A PCR array was used to screen the expression of 84 key genes involved in cell migration. (-)-DHMEQ inhibited antigen/IgE-induced NF-κB activation and expressions of its target genes such as IL-6 and TNF-α. (-)-DHMEQ was found to inhibit in vitro invasion toward the antigen without any toxicity. We then looked for NF-κB-dependent genes that would be important for mast cell invasion using the PCR array. (-)-DHMEQ was found to lower the expression of matrix metalloproteinase (MMP)-2. The MMP inhibitor GM6001 also inhibited cellular invasion toward the antigen. These effects of (-)-DHMEQ were obtained in both RBL-2H3 cells and BMMCs. These findings indicate that (-)-DHMEQ suppressed mast cell migration via the inhibition of NF-κB-regulated MMP-2 expression.


Effects of exercise on leukocyte death: prevention by hydrolyzed whey protein enriched with glutamine dipeptide.
Lymphocyte and neutrophil death induced by exercise and the role of hydrolyzed whey protein enriched with glutamine dipeptide (Gln) supplementation was investigated. Nine triathletes performed two exhaustive exercise trials with a 1-week interval in a randomized, double blind, crossover protocol. Thirty minutes before treadmill exhaustive exercise at variable speeds in an inclination of 1% the subjects ingested 50 g of maltodextrin (placebo) or 50 g of maltodextrin plus 4 tablets of 700 mg of hydrolyzed whey protein enriched with 175 mg of glutamine dipeptide dissolved in 250 mL water. Cell viability, DNA fragmentation, mitochondrial transmembrane potential and production of reactive oxygen species (ROS) were determined in lymphocytes and neutrophils. Exhaustive exercise decreased viable lymphocytes but had no effect on neutrophils. A 2.2-fold increase in the proportion of lymphocytes and neutrophils with depolarized mitochondria was observed after exhaustive exercise. Supplementation of maltodextrin plus Gln (MGln) prevented the loss of lymphocyte membrane integrity and the mitochondrial membrane depolarization induced by exercise. Exercise caused an increase in ROS production by neutrophils, whereas supplementation of MGln had no additional effect. MGln supplementation partially prevented lymphocyte apoptosis induced by exhaustive exercise possibly by a protective effect on mitochondrial function.


Bacterial and human peptidylarginine deiminases: targets for inhibiting the autoimmune response in rheumatoid arthritis?
Peptidylarginine deiminases (PADs) convert arginine within a peptide (peptidylarginine) into peptidylcitrulline. Citrullination by human PADs is important in normal physiology and inflammation. Porphyromonas gingivalis, a major pathogen in periodontitis, is the only prokaryote described to possess PAD. P. gingivalis infection may generate citrullinated peptides, which trigger anti-citrullinated peptide antibodies. In susceptible individuals, host protein citrullination by human PADs in the joint probably perpetuates antibody formation, paving the way for the development of chronic arthritis. Blockades of bacterial and human PADs may act as powerful novel therapies by inhibiting the generation of the antigens that trigger and sustain autoimmunity in rheumatoid arthritis.


An overview of the European Health Examination Survey Pilot Joint Action.
Health Examination Surveys (HESs) can provide essential information on the health and health determinants of a population, which is not available from other data sources. Nevertheless, only some European countries have systems of national HESs. A study conducted in 2006-2008 concluded that it is feasible to organize national HESs using standardized measurement procedures in nearly all EU countries. The feasibility study also outlined a structure for a European Health Examination Survey (EHES), which is a collaboration to organize standardized HESs in countries across Europe.To facilitate setting up national surveys and to gain experience in applying the EHES methods in different cultures, EHES Joint Action (2010-2011) planned and piloted standardized HESs in the working age population in 12 countries. This included countries with earlier national HESs and countries which were planning their first national HES. The core measurements included in all surveys were weight, height, waist circumference and blood pressure, and blood samples were taken to measure lipid profiles and glucose or glycated haemoglobin (HbA1c). These are modifiable determinants of major chronic diseases not identified in health interview surveys. There was a questionnaire to complement the data on the examination measurements. Evaluation of the pilot surveys was based on review of national manuals and evaluation reports of survey organizers; observations and discussions of survey procedures during site visits and training seminars; and other communication with the survey organizers. Despite unavoidable differences in the ways HESs are organized in the various countries, high quality and comparability of the data seems achievable. The biggest challenge in each country was obtaining high participation rate. Most of the pilot countries are now ready to start their full-size national HES, and six of them have already started. The EHES Pilot Project has set up the structure for obtaining comparable high quality health indicators on health and important modifiable risk factors of major non-communicable diseases from the European countries. The European Union is now in a key position to make this structure sustainable. The EHES core survey can be expanded to cover other measurements.


The functional variant rs334558 of GSK3B is associated with remission in patients with depressive disorders.
GSK3B and AKT1 genes have been implicated in the pathogenesis of a number of psychiatric and neurological disorders. Furthermore, their genetic variants are associated with response to antidepressant pharmacotherapy. As the evidence is still incomplete and inconsistent, continuing efforts to investigate the role of these two genes in the pathogenesis and treatment of brain disorders is necessary. The aim of our study was thus to evaluate the association of variants of these two genes with depressive disorders and drug treatment response. In the present study, 222 patients with a depressive disorder who underwent pharmacological antidepressant treatment were divided into remitters and non-remitters following a 28-day course of pharmacotherapy. The association of a depressive disorder and remission rates with polymorphisms rs334558 in the GSK3B gene and rs1130214 and rs3730358 in the AKT1 gene was evaluated with a chi-square test. Neither of the studied genetic variants was associated with a depressive disorder. Furthermore, frequencies of alleles and genotypes for rs1130214 and rs3730358 were not different in the groups of remitters and non-remitters. However, the activating allele T of the functional polymorphism rs334558 was significantly associated with remission, when all types of antidepressant drugs were included. This association continued as a trend when only patients taking selective serotonin reuptake inhibitors were considered. The present study provides support that the functional polymorphism rs334558 of GSK3B may play a role as a useful genetic and pharmacogenetic biomarker in the framework of personalized medicine approach.


Ccbe1 regulates Vegfc-mediated induction of Vegfr3 signaling during embryonic lymphangiogenesis.
The VEGFC/VEGFR3 signaling pathway is essential for lymphangiogenesis (the formation of lymphatic vessels from pre-existing vasculature) during embryonic development, tissue regeneration and tumor progression. The recently identified secreted protein CCBE1 is indispensible for lymphangiogenesis during development. The role of CCBE1 orthologs is highly conserved in zebrafish, mice and humans with mutations in CCBE1 causing generalized lymphatic dysplasia and lymphedema (Hennekam syndrome). To date, the mechanism by which CCBE1 acts remains unknown. Here, we find that ccbe1 genetically interacts with both vegfc and vegfr3 in zebrafish. In the embryo, phenotypes driven by increased Vegfc are suppressed in the absence of Ccbe1, and Vegfc-driven sprouting is enhanced by local Ccbe1 overexpression. Moreover, Vegfc- and Vegfr3-dependent Erk signaling is impaired in the absence of Ccbe1. Finally, CCBE1 is capable of upregulating the levels of fully processed, mature VEGFC in vitro and the overexpression of mature VEGFC rescues ccbe1 loss-of-function phenotypes in zebrafish. Taken together, these data identify Ccbe1 as a crucial component of the Vegfc/Vegfr3 pathway in the embryo.


Alternatively spliced isoforms of IL-32 differentially influence cell death pathways in cancer cell lines.
Alternative splicing is a biological mechanism that enables the synthesis of several isoforms with different or even opposite functions. This process must be tightly regulated to prevent unwanted isoform expression favoring pathological processes. Some isoforms of interleukin 32 (IL-32) are reported to be more potent in inducing inflammation, however the role in cell death remains to be investigated. This study demonstrates that IL-32γ and IL-32β can induce caspase-8-dependent cell death whereas this was not observed for IL-32α. Overexpression of IL-32β or IL-32γ but not IL-32α, resulted in enhanced expression of the survival cytokine IL-8. Furthermore, restoring the IL-8 signaling pathway by overexpressing CXCR1 in HEK293 cells, rescued IL-32β but not IL-32γ-induced cell death. Interestingly, IL-32γ was able to downregulate CXCR1 and thereby induce cell death. Subsequent studies into the role of IL-32 in thyroid cancer (TC) revealed that several IL-32 isoforms, IL-8, and CXCR1 are expressed in TC cell lines and specimens. Remarkably, TC cell lines were found to produce high concentrations of IL-8, indicating an important role for IL-8 in the survival-signaling pathway in these cells. Intriguingly, a significant correlation between the IL-8 receptor CXCR1 and IL-32γ was observed in TC specimens, while this was not observed for the other IL-32 splice variants. Blocking IL-32 alternative splicing by Isoginkgetin resulted in predominant expression of IL-32γ splice variants and cell death in TC cell lines. All together, modulation of IL-32 alternative splicing could represent a novel strategy for the treatment of malignancies, in particular thyroid cancer.


Antibiotic resistance prediction for Mycobacterium tuberculosis from genome sequence data with Mykrobe.
Two billion people are infected with Mycobacterium tuberculosis, leading to 10 million new cases of active tuberculosis and 1.5 million deaths annually. Universal access to drug susceptibility testing (DST) has become a World Health Organization priority. We previously developed a software tool, Mykrobe predictor, which provided offline species identification and drug resistance predictions for M. tuberculosis from whole genome sequencing (WGS) data. Performance was insufficient to support the use of WGS as an alternative to conventional phenotype-based DST, due to mutation catalogue limitations. Here we present a new tool, Mykrobe, which provides the same functionality based on a new software implementation. Improvements include i) an updated mutation catalogue giving greater sensitivity to detect pyrazinamide resistance, ii) support for user-defined resistance catalogues, iii) improved identification of non-tuberculous mycobacterial species, and iv) an updated statistical model for Oxford Nanopore Technologies sequencing data. Mykrobe is released under MIT license at https://github.com/mykrobe-tools/mykrobe. We incorporate mutation catalogues from the CRyPTIC consortium et al. (2018) and from Walker et al. (2015), and make improvements based on performance on an initial set of 3206 and an independent set of 5845 M. tuberculosis Illumina sequences. To give estimates of error rates, we use a prospectively collected dataset of 4362 M. tuberculosis isolates. Using culture based DST as the reference, we estimate Mykrobe to be 100%, 95%, 82%, 99% sensitive and 99%, 100%, 99%, 99% specific for rifampicin, isoniazid, pyrazinamide and ethambutol resistance prediction respectively. We benchmark against four other tools on 10207 (=5845+4362) samples, and also show that Mykrobe gives concordant results with nanopore data. We measure the ability of Mykrobe-based DST to guide personalized therapeutic regimen design in the context of complex drug susceptibility profiles, showing 94% concordance of implied regimen with that driven by phenotypic DST, higher than all other benchmarked tools.


A prospective study of serum 25-hydroxyvitamin D levels, blood pressure, and incident hypertension in postmenopausal women.
In randomized trials, the effect of vitamin D supplementation on blood pressure has been equivocal, while most prospective cohort studies have shown that the risk of incident hypertension is lower in people with higher levels of 25-hydroxyvitamin D (25(OH)D). The authors examined the association between levels of 25(OH)D and changes in blood pressure and incident hypertension in 4,863 postmenopausal women recruited into the Women's Health Initiative between 1993 and 1998. Over 7 years, there were no significant differences in the adjusted mean change in systolic or diastolic blood pressure by quartile of 25(OH)D. The covariate-adjusted risk of incident hypertension was slightly lower in the upper 3 quartiles of 25(OH)D compared with the lowest quartile, but this was statistically significant only in the third quartile (hazard ratio = 0.67, 95% confidence interval: 0.46, 0.96). There was no significant linear or nonlinear trend in the risk of incident hypertension by untransformed or log-transformed continuous values of 25(OH)D. In postmenopausal women in this study, serum levels of 25(OH)D were not related to changes in blood pressure, and evidence for an association with lower risk of incident hypertension was weak.


Brain Perivascular Macrophages Initiate the Neurovascular Dysfunction of Alzheimer Aβ Peptides.
Increasing evidence indicates that alterations of the cerebral microcirculation may play a role in Alzheimer disease, the leading cause of late-life dementia. The amyloid-β peptide (Aβ), a key pathogenic factor in Alzheimer disease, induces profound alterations in neurovascular regulation through the innate immunity receptor CD36 (cluster of differentiation 36), which, in turn, activates a Nox2-containing NADPH oxidase, leading to cerebrovascular oxidative stress. Brain perivascular macrophages (PVM) located in the perivascular space, a major site of brain Aβ collection and clearance, are juxtaposed to the wall of intracerebral resistance vessels and are a powerful source of reactive oxygen species. We tested the hypothesis that PVM are the main source of reactive oxygen species responsible for the cerebrovascular actions of Aβ and that CD36 and Nox2 in PVM are the molecular substrates of the effect. Selective depletion of PVM using intracerebroventricular injection of clodronate abrogates the reactive oxygen species production and cerebrovascular dysfunction induced by Aβ applied directly to the cerebral cortex, administered intravascularly, or overproduced in the brain of transgenic mice expressing mutated forms of the amyloid precursor protein (Tg2576 mice). In addition, using bone marrow chimeras, we demonstrate that PVM are the cells expressing CD36 and Nox2 responsible for the dysfunction. Thus, deletion of CD36 or Nox2 from PVM abrogates the deleterious vascular effects of Aβ, whereas wild-type PVM reconstitute the vascular dysfunction in CD36-null mice. The data identify PVM as a previously unrecognized effector of the damaging neurovascular actions of Aβ and unveil a new mechanism by which brain-resident innate immune cells and their receptors may contribute to the pathobiology of Alzheimer disease.


Helicobacter pylori CagA-mediated IL-8 induction in gastric epithelial cells is cholesterol-dependent and requires the C-terminal tyrosine phosphorylation-containing domain.
Upon infection of the gastric epithelial cells, the Helicobacter pylori cytotoxin-associated gene A (CagA) virulence protein is injected into the epithelial cells via the type IV secretion system (TFSS), which is dependent on cholesterol. Translocated CagA is targeted by the membrane-recruited c-Src family kinases in which a tyrosine residue in the Glu-Pro-Ile-Tyr-Ala (EPIYA)-repeat region, which can be phosphorylated, induces cellular responses, including interleukin-8 (IL-8) secretion and hummingbird phenotype formation. In this study, we explored the role of EPIYA-containing C-terminal domain (CTD) in CagA tethering to the membrane lipid rafts and in IL-8 activity. We found that disruption of the lipid rafts reduced the level of CagA translocation/phosphorylation as well as CagA-mediated IL-8 secretion. By CagA truncated mutagenesis, we identified that the CTD, rather than the N-terminal domain, was responsible for CagA tethering to the plasma membrane and association with detergent-resistant membranes, leading to CagA-induced IL-8 promoter activity. Our results suggest that CagA CTD-containing EPIYAs directly interact with cholesterol-rich microdomains that induce efficient IL-8 secretion in the epithelial cells.


Evaluation of efficacy of probiotics in prevention of candida colonization in a PICU-a randomized controlled trial.
To evaluate the efficacy of probiotics in prevention of Candida colonization in a PICU. Prospective double blinded, randomized controlled trial. PICU of a tertiary care teaching hospital in north India. One hundred fifty children (106 boys, 44 girls), 3 months to 12 yrs old, on broad spectrum antibiotics for at least 48 hrs were randomized using computer-generated random numbers to receive probiotic mix (EUGI) (n = 75) or placebo (n = 75). Patients received one sachet twice a day of either probiotics or placebo for 7 days. Probiotics contained Lactobacillus acidophillus, Lactobacillus rhamnosum, Bifidobacterium longum, Bifidobacterium bifidum, Saccharomyces boulardi, Saccharomyces thermophilus, fructo-oligosaccharides; and placebo-contained lactose packed in similar-looking sachets. Rectal swabs for fungal culture were taken at day 0, 7, and 14 of enrollment. Primary outcome measure was prevalence of rectal colonization with Candida on day 14 postenrollment; secondary outcomes were growth of Candida in urine (candiduria) and blood (candidemia). Patients were followed until completion of 14 days study period or death of patient. Demographic and clinical variables were comparable in two groups. Prevalence of Candida colonization on day 0 was similar (15 of 75) in both the groups. On day 7, 27.9% (19 of 68) patients in the probiotic group and 42.6% (29 of 68) patients in the placebo group were colonized (relative risk 0.65; 95% confidence interval 0.41-1.05; p = 0.07), whereas, on day 14, colonization was observed in 31.3% (21 of 67) patients in the probiotic group and 50% (34 of 68) in the placebo group (relative risk 0.63; 95% confidence interval 0.41-0.96; p = 0.02). Thus, the relative reduction in prevalence of Candida colonization on day 7 and 14 in the probiotic group was 34.5% and 37.2%, respectively. The increase in number of colonized patients from day 0 to 7 and day 0 to 14 was significant in the placebo group (p = 0.004 and 0.001, respectively) but not in the probiotic group (p = 0.30 and 0.19, respectively; McNemar test). Candiduria was significantly less common in the probiotic group than in the placebo group (17.3% vs. 37.3%; relative risk 0.46; 95% confidence interval 0.26-0.82; p = 0.006). However, prevalence of candidemia did not differ significantly in two groups (1.6% in the probiotic group vs. 6.35% in placebo group; relative risk 0.46; 95% confidence interval 0.08-2.74; p = 0.39). Supplementation with probiotics could be a potential strategy to reduce gastrointestinal Candida colonization and candiduria in critically ill children receiving broad spectrum antibiotics.


Intense sympathetic stimulation releases neuropeptide Y but fails to evoke sustained coronary vasoconstriction in dogs.
We determined whether a 3-minute period of intense cardiac sympathetic stimulation, which is known to release neuropeptide Y (NPY), elicits a sustained poststimulatory coronary vasoconstriction in anesthetized dogs that had received propranolol. We also periodically measured the cardiac chronotropic responses to test vagal stimulations; these responses served as an index of the neuronal release of NPY. In a group of 11 animals, the coronary vascular resistance increased by 14 +/- 4% during the sympathetic stimulation. After cessation of stimulation, however, coronary vascular resistance returned rapidly to its control value. The cardiac responses to the test vagal stimuli were attenuated by approximately 40% after cessation of sympathetic stimulation, and this inhibitory effect persisted for approximately 60 minutes. In a second group of eight dogs, we determined whether the intense sympathetic stimulation potentiates the coronary vascular responses to exogenous norepinephrine (NE). Before sympathetic stimulation, standard intracoronary infusions of NE increased coronary vascular resistance by 14 +/- 2%. Intense antecedent sympathetic stimulation did not alter the coronary vascular responses to subsequent NE infusions. However, the chronotropic responses to test vagal stimuli were initially attenuated by approximately 30%, and this inhibitory effect persisted for approximately 1 hour. In a third group of four dogs, we found that exogenous NPY significantly potentiated the coronary vasoconstriction evoked by NE infusions. The coronary vascular responses to combined infusions of NE and NPY were consistently greater (by approximately 13%) than the sum of the responses to these substances when they were infused separately. We conclude that, even though sufficient NPY appears to be released from the sympathetic nerve endings to inhibit vagal neurotransmission, the quantity of NPY released into the coronary blood vessels under the conditions of our experiments appears to be insufficient either to elicit a sustained coronary vasoconstriction or to potentiate the vasoconstrictor effects of intracoronary NE infusions.


Inhibition of Ly-6A antigen expression prevents T cell activation.
Antisense oligonucleotides complementary to the 5' end of the mRNA encoding the Ly-6A protein were used to block the expression of that protein. Using this approach we could inhibit the expression of Ly-6A by 60-80% in antigen-primed lymph node (LN) T cells as well as in the D10 T cell clone. Inhibition of Ly-6 expression resulted in the inability to restimulate in vitro, antigen-primed T cells. It also blocked the activation of normal spleen cells by Con A, monoclonal antibody (mAb) to CD3, and mAb to Ly-6. In contrast, stimulation of normal spleen cells with the pharmacological agents PMA + ionomycin were unaffected by the inhibition of Ly-6 expression. Similar results were obtained with the D10 T cell clone; stimulation with Con A + interleukin 1 (IL-1), antigen-presenting cells (APC), or the clonotypic antibody + IL-1 was greatly reduced in the presence of antisense oligonucleotides to Ly-6. Stimulation with PMA + ionomycin was again unaffected. We also studied the effect of antisense oligonucleotides on stimulation of preactivated D10 cells. Preactivation of D10 cells with Con A + IL-1 renders them receptive to secondary stimulation by other lymphokines. In this case, antisense oligonucleotides to Ly-6 had no effect on secondary activation with IL-2, IL-4 + IL-1, or PMA + ionomycin. We conclude from these studies that Ly-6 expression is required for T cell receptor (TCR)-mediated T cell activation.


Ultrathin epitaxial Bi film growth on 2D HfTe2template.
Among ultrathin monoelemental two-dimensional (2D) materials, bismuthene, the single layer of heavier group-VΑ element bismuth (Bi), has been predicted to have large non trivial gap. Here, we demonstrate the growth of Bi films by molecular beam epitaxy on 2D-HfTe2template. At the initial stage of Bi deposition (1-2 bilayers, BL), both the pseudocubic Bi(110) and the hexagonal Bi(111) phases are formed. When reaching 3 BL Bi, a transformation to pure hexagonal Bi(111) occurs. The electronic band structure of 3 BL Bi(111) films was measured by angle-resolved photoemission spectroscopy showing very good matching with the density functional theory band structure calculations of 3 BL free standing Bi(111). The grown Bi(111) thin film was capped with a protective Al2O3layer and its stability under ambient conditions, necessary for practical applications and device fabrication, was confirmed by x-ray photoelectron spectroscopy and Raman spectroscopy.


Shadow removal using intensity surfaces and texture anchor points.
Removal of shadows from a single image is a challenging problem. Producing a high-quality shadow-free image which is indistinguishable from a reproduction of a true shadow-free scene is even more difficult. Shadows in images are typically affected by several phenomena in the scene, including physical phenomena such as lighting conditions, type and behavior of shadowed surfaces, occluding objects, etc. Additionally, shadow regions may undergo post-acquisition image processing transformations, e.g., contrast enhancement, which may introduce noticeable artifacts in the shadow-free images. We argue that the assumptions introduced in most studies arise from the complexity of the problem of shadow removal from a single image and limit the class of shadow images which can be handled by these methods. The purpose of this paper is twofold: First, it provides a comprehensive survey of the problems and challenges which may occur when removing shadows from a single image. In the second part of the paper, we present our framework for shadow removal, in which we attempt to overcome some of the fundamental problems described in the first part of the paper. Experimental results demonstrating the capabilities of our algorithm are presented.


Total parenteral nutrition in treatment of patients with inflammatory bowel disease.
Patients with Inflammatory Bowel Disease (IBD) are exposed to nutritional risk. Malnutrition in Crohn's Disease (CD) and to a somewhat lesser in Ulcerative Colitis (UC) is very frequent. Depending on the severity of the disease, weight loss has been reported in 65% to 76% of those with CD and in 18% to 62% of those with UC. The role of Total Parenteral Nutrition (TPN) is essential in very severe cases where enteral nutrition is not tolerated or standard drug therapy is not effective. Nutritional therapy is important for the correction of nutritional deficiency, especially in cases where elective surgical treatment is required. This study examined the effects of preoperative TPN administration in patients with IBD. Since 1990, 29 pts, 13 (44.8%) male and 16 (55.2%) female with severe IBD; 16 (55.2%) with UC and 13 (44.8%) with CD were treated with TPN in our department in the preoperative period. Evaluation of this group was compared with a group which was not treated with TPN preoperatively: 61 pts, 34 (55.7%) male, 27 (44.3%) female; 50 (82%) with CD, 11 (18%) with UC. Evaluation of this group was compared with the group of patients who were subjected to surgical procedure without prior TPN administration, in total a number of 61, of whom 34 (55.7%) were male and 27 (44.3%) female patients. In this group, the number of patients with CD and UC was 50 (82%) and 11(18%) respectively. During the course of the study, the following parameters were examined: Body mass index (BMI), Disease Activity Index (CDAI/AI), laboratory parameters and the number of hospital days. The parameters were analysed before the surgical intervention, and one week and six months after the surgical intervention. The duration of the application of TPN was 12.5 +/- 5 days. The analysis of these parameters has shown that there is no statistically significant difference in the number of hospital days in both groups, which was 18.9 +/- 8.9 in the intervention group and 18.9 +/- 6.5 days in the control group, p = 0.9808, but analysis of the Disease Activity (CDAI/AI) has shown that patients who were on TPN were in a more severe stage of the disease. In the TPN treated group of patients none, 0 pts (0%), of the pts had no Index of Activity, 1.7 pts (24.1%) had Index 2 and 22 (75.9%) had Index 3. In the other group 3 pts (4.9%) had Index 1; 39 pts (63.9%) had Index 2 and 19 pts (31.2%) had Index of Activity 3. During the monitoring period of six months the activity of the disease was lower in patients treated with TPN. The BMI in the group of patients treated with TPN was lower in both sexes. It was lower than 18.4 in 7 males and 5 females; between 18.4 and 19.9 in 2 males and 4 females; between 19.9 and 25 in 3 males and 6 females; between 25 and 19.9 in 1 male and 1 female; there were no pts with BMI higher than 29.9. In the control non-TPN group, 5 males and 3 females had BMI < or = 18.4; 2 males and 5 females had between 18.4 and 19.9; 23 males and 13 females had between 19.9 and 25; 3 males and 5 females had between 25 and 29.9 and 1 male and 1 female had BMI higher than 29.9. The BMI had an increasing trend in the postoperative period in both groups. The laboratory parameters that we examined were: number of erythrocytes and sedimentation, number of leukocytes, haemoglobin levels, total number of lymphocytes, albumin level, C-reactive protein, orosomucoid, electrolytes; sodium, potassium, calcium and serum iron, the enzymes (AP, AST and ALT); level of bilirubin and urea and creatinine. The results were analysed using the Tukey honest significant difference test (HSD), ANOVA and Student t-test. Statistically significant differences were observed between preoperative and postoperative period in both groups. The intergroup comparison showed significant differences in the level of albumin, AST, ALT and bilirubin.


Biocompatible mannosylated endosomal-escape nanoparticles enhance selective delivery of short nucleotide sequences to tumor associated macrophages.
Tumor associated macrophages (TAMs) can modify the tumor microenvironment to create a pro-tumor niche. Manipulation of the TAM phenotype is a novel, potential therapeutic approach to engage anti-cancer immunity. siRNA is a molecular tool for knockdown of specific mRNAs that is tunable in both strength and duration. The use of siRNA to reprogram TAMs to adopt an immunogenic, anti-tumor phenotype is an attractive alternative to ablation of this cell population. One current difficulty with this approach is that TAMs are difficult to specifically target and transfect. We report here successful utilization of novel mannosylated polymer nanoparticles (MnNP) that are capable of escaping the endosomal compartment to deliver siRNA to TAMs in vitro and in vivo. Transfection with MnNP-siRNA complexes did not significantly decrease TAM cell membrane integrity in culture, nor did it create adverse kidney or liver function in mice, even at repeated doses of 5 mg kg(-1). Furthermore, MnNP effectively delivers labeled nucleotides to TAMs in mice with primary mammary tumors. We also confirmed TAM targeting in the solid tumors disseminated throughout the peritoneum of ovarian tumor bearing mice following injection of fluorescently labeled MnNP-nucleotide complexes into the peritoneum. Finally, we show enhanced uptake of MnNP in lung metastasis associated macrophages compared to untargeted particles when using an intubation delivery method. In summary, we have shown that MnNP specifically and effectively deliver siRNA to TAMs in vivo.


[Proposals for the reform of public health services in Catalonia].
In the year 2004 the government of Catalonia undertook a process to reform its public health services. In this context, it created a working groupinvolving experts from diverse backgrounds to analyse the reforms to be undertaken, the Scientific Committee for the Reform of Public Health in Catalonia. Its members produced eight documents on specific aspects of public health, from which a global report of the Committee was compiled by the end of 2005. This paper makes a synthesis of their production, and includes as an annex their recommendations and proposals. Public health policies should be structured around three main goal: the reduction of health inequalities, the control and removal of social and environmental risks, and effective improvements in quality of life. To reach them, common criteria are defined as main directions. These are based in favouring decentralization of public health services and their administration, linking public health activities with health care services, designing interventions with a population perspective, and reinforcing cross-sectional implications of public health. The work of this Committee is produced in the context of an international debate on the future of public health services and the disproportion between its contribution to health and well being and its resources and visibility. The Committee produced proposals and recommendations which can he grouped in five facets: consolidating a solid and coherent system, developing an organizational reform, defining a port-folio of services, adopting improvements in management, and taking into account cross sectional aspects relating to public health.


Simultaneous quantification of cannabinoids and metabolites in oral fluid by two-dimensional gas chromatography mass spectrometry.
Development and validation of a method for simultaneous identification and quantification of Delta9-tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), and metabolites 11-hydroxy-THC (11-OH-THC) and 11-nor-9-carboxy-THC (THCCOOH) in oral fluid. Simultaneous analysis was problematic due to different physicochemical characteristics and concentration ranges. Neutral analytes, such as THC and CBD, are present in ng/mL, rather than pg/mL concentrations, as observed for the acidic THCCOOH biomarker in oral fluid. THCCOOH is not present in cannabis smoke, definitively differentiating cannabis use from passive smoke exposure. THC, 11-OH-THC, THCCOOH, CBD, and CBN quantification was achieved in a single oral fluid specimen collected with the Quantisal device. One mL oral fluid/buffer solution (0.25 mL oral fluid and 0.75 mL buffer) was applied to conditioned CEREX Polycrom THC solid-phase extraction (SPE) columns. After washing, THC, 11-OH-THC, CBD, and CBN were eluted with hexane/acetone/ethyl acetate (60:30:20, v/v/v), derivatized with N,O-bis-(trimethylsilyl)trifluoroacetamide and quantified by two-dimensional gas chromatography electron ionization mass spectrometry (2D-GCMS) with cold trapping. Acidic THCCOOH was separately eluted with hexane/ethyl acetate/acetic acid (75:25:2.5, v/v/v), derivatized with trifluoroacetic anhydride and hexafluoroisopropanol, and quantified by the more sensitive 2D-GCMS-electron capture negative chemical ionization (NCI-MS). Linearity was 0.5-50 ng/mL for THC, 11-OH-THC, CBD and 1-50 ng/mL for CBN. The linear dynamic range for THCCOOH was 7.5-500 pg/mL. Intra- and inter-assay imprecision as percent RSD at three concentrations across the linear dynamic range were 0.3-6.6%. Analytical recovery was within 13.8% of target. This new SPE 2D-GCMS assay achieved efficient quantification of five cannabinoids in oral fluid, including pg/mL concentrations of THCCOOH by combining differential elution, 2D-GCMS with electron ionization and negative chemical ionization. This method will be applied to quantification of cannabinoids in oral fluid specimens from individuals participating in controlled cannabis and Sativex (50% THC and 50% CBD) administration studies, and during cannabis withdrawal.


Cellular therapy to target neuroinflammation in amyotrophic lateral sclerosis.
Neurodegenerative disorders are characterized by the selective vulnerability and progressive loss of discrete neuronal populations. Non-neuronal cells appear to significantly contribute to neuronal loss in diseases such as amyotrophic lateral sclerosis (ALS), Parkinson, and Alzheimer's disease. In ALS, there is deterioration of motor neurons in the cortex, brainstem, and spinal cord, which control voluntary muscle groups. This results in muscle wasting, paralysis, and death. Neuroinflammation, characterized by the appearance of reactive astrocytes and microglia as well as macrophage and T-lymphocyte infiltration, appears to be highly involved in the disease pathogenesis, highlighting the involvement of non-neuronal cells in neurodegeneration. There appears to be cross-talk between motor neurons, astrocytes, and immune cells, including microglia and T-lymphocytes, which are subsequently activated. Currently, effective therapies for ALS are lacking; however, the non-cell autonomous nature of ALS may indicate potential therapeutic targets. Here, we review the mechanisms of action of astrocytes, microglia, and T-lymphocytes in the nervous system in health and during the pathogenesis of ALS. We also evaluate the therapeutic potential of these cellular populations, after transplantation into ALS patients and animal models of the disease, in modulating the environment surrounding motor neurons from pro-inflammatory to neuroprotective. We also thoroughly discuss the recent advances made in the field and caveats that need to be overcome for clinical translation of cell therapies aimed at modulating non-cell autonomous events to preserve remaining motor neurons in patients.


The T cell and NK cell immune response to exercise.
Exercise elicits an increase in the numbers of circulating lymphocytes and lymphocyte subsets (including NK cells) which is followed by a decrease in the numbers of cells during recovery from exercise; this lymphocytopenia appears to be due to a decrease in the percentage of type 1 T cells and NK cells in the circulation at this time. A decrease in mitogen-stimulated T cell proliferation and T cell production of IL-2 and IFN-gamma is reported immediately after acute, intensive exercise. NK cell cytolytic activity per cell (NKCA) does not appear to change much after exercise unless the bout was prolonged, intense and stressful, in which case NKCA can be depressed for several hours. Resting immune function is not very different in athletes compared with non-athletes. However, periods of intensified training in already well trained athletes can result in a depression of immunity in the resting state which may be due to the cumulative effects of repeated bouts of intense exercise with the consequent elevation of stress hormones, particularly cortisol and anti-inflammatory cytokines (e.g. IL-6, IL-10, IL-Ira) causing temporary inhibition of type 1 T cell cytokine production with a relative dampening of the type I (cell-mediated) response.


Predictors of lost time from work among nursing personnel who sought treatment for back pain.
To examine possible predictors of lost workdays among nurses and nurses' aides who sought treatment for work-related back pain. Nursing staff employed at a tertiary care medical center over a 13-year time period (1994 through 2006). We used existing data from clinic surveys administered to nursing personnel during their initial treatment visit to the hospital's occupational health clinic. Predictors of losing ≤ 7 and ≥ 8 workdays was examined. 589 of 708 (83%) nursing personnel with complaints of work-related back pain completed the survey, with 31% resulting in lost workdays. Experiencing sudden onset of pain (RR:1.9; 95% CI: 1.1, 3.1), a combination of severe pain with numbness and tingling in the back/legs (RR: 7.4; 95% CI: 2.9, 18.6), severe pain only (RR: 4.4; 95% CI: 1.8, 11.1), numbness and tingling in the back/legs only (RR: 3.5; 95% CI: 1.0, 12.2), and working < 5 years at the hospital (RR: 2.3; 95% CI: 1.2, 4.7) were predictive of losing ≥ 8 workdays. Job title, work demands, work conflicts, and most psychosocial factors were not predictive. Severe pain, neurologic symptoms and sudden onset of pain were predictive of delayed return-to-work; however, these symptoms alone should not be considered indicators of poor outcomes given that most workers who reported these symptoms returned to work in less than 8~days. Among these health care workers, lost workdays appear to be related to more severe pathology rather than workplace characteristics.


Peer volunteers improve long-term maintenance of physical activity with older adults: a randomized controlled trial.
Using peer volunteers as delivery agents may improve translation of evidence-based physical activity promotion programs for older adults. This study examined whether tailored support from older peer volunteers could improve initiation and long-term maintenance of physical activity behavior. Participants were randomized to 2 16-week, group-based programs: (1) peer-delivered, theory-based support for physical activity behavior change; or (2) an intervention typically available in community settings (basic education, gym membership, and pedometer for self-monitoring), attention-matched with health education. Moderate-to-vigorous physical activity (MVPA) was assessed via daily self-report logs at baseline, at the end of the intervention (16 weeks), and at follow-up (18 months), with accelerometry validation (RT3) in a random subsample. Seven peer volunteers and 81 sedentary adults were recruited. Retention at the end of the trial was 85% and follow-up at 18 months was 61%. Using intent-to-treat analyses, at 16 weeks, both groups had similar significant improvements in MVPA. At 18 months, the group supplemented with peer support had significantly more MVPA. Trained peer volunteers may enhance long-term maintenance of physical activity gains from a community-based intervention. This approach has great potential to be adapted and delivered inexpensively in community settings.


Swedish fathers contemplate the difficulties they face in parenthood.
The aim was to explore what concerns Swedish fathers had about parenting difficulties at two months after the birth of their baby. Self-report questionnaires were used and data were analyzed with mixed methods. Thirty percent of the 827 fathers reported concerns about the difficulties of parenthood. The theme 'Managing the demands of being a father' emerged and was based on concerns about how to raise the baby, having enough money, health issues, lack of time and finding balance in the new family pattern. Financial worries, feeling less positive about expecting a baby, and self-reported poor emotional health were related to fathers who perceived parenthood as difficult. Experienced fathers as well as new fathers expressed similar concerns about parenthood. Preparation classes for reassurance and skills coaching about child raising may provide important support for fathers. This is especially important for fathers who may have poor emotional health or who may not be feeling positive about expecting a baby. Policy-makers and health care providers should recognize that offering support for all fathers benefits not only men, but also their children, and their partners and can help encourage egalitarian practices at home and work.


Characterization of NGF, trkA (NGFR) , and p75 (NTR) in Retina of Mice Lacking Reelin Glycoprotein.
Both Reelin and Nerve Growth Factor (NGF) exert crucial roles in retinal development. Retinogenesis is severely impaired in E-reeler mice, a model of Reelin deficiency showing specific Green Fluorescent Protein expression in Rod Bipolar Cells (RBCs). Since no data are available on Reelin and NGF cross-talk, NGF and trkA(NGFR)/ p75(NTR) expression was investigated in retinas from E-reeler versus control mice, by confocal microscopy, Western blotting, and real time PCR analysis. A scattered increase of NGF protein was observed in the Ganglion Cell Layer and more pronounced in the Inner Nuclear Layer (INL). A selective increase of p75(NTR) was detected in most of RBCs and in other cell subtypes of INL. On the contrary, a slight trend towards a decrease was detected for trkA(NGFR), albeit not significant. Confocal data were validated by Western blot and real time PCR. Finally, the decreased trkA(NGFR)/ p75(NTR) ratio, representative of p75(NTR) increase, significantly correlated with E-reeler versus E-control. These data indicate that NGF-trkA(NGFR)/ p75(NTR) is affected in E-reeler retina and that p75(NTR) might represent the main NGF receptor involved in the process. This first NGF-trkA(NGFR)/ p75(NTR) characterization suggests that E-reeler might be suitable for exploring Reelin-NGF cross-talk, representing an additional information source in those pathologies characterized by retinal degeneration.


Relevance of the light line in planar photonic crystal waveguides with weak vertical confinement.
The concept of the so-called light line is a useful tool to distinguish between guided and non-guided modes in dielectric slab waveguides. Also for more complicated structures with 2D mode confinement, the light lines can often be used to divide a dispersion diagram into a region of a non-guided continuum of modes, a region of discrete guided modes and a forbidden region, where no propagating modes can exist. However, whether or not the light line is a concept of practical relevance depends on the geometry of the structure. This fact is sometimes ignored. For instance, in the literature on photonic crystal waveguides, it is often argued that substrate-type photonic crystal waveguides with a weak vertical confinement are inherently lossy, since the entire bandgap including the line defect modes is typically located above the light line of the substrate. The purpose of this article is to illustrate that this argument is inaccurate and to provide guidelines on how an improved light line concept can be constructed.


Do Vestibular-Evoked Myogenic Potential Abnormalities in Patients with Cochlear Implant Only Reflect Saccular Dysfunction?
The objective of this study was to explore the usefulness of 1000-Hz tone burst (TB) stimuli for detecting cervical vestibular-evoked myogenic potential (cVEMP) abnormalities in patients with a cochlear implant (CI). Thirty asymptomatic patients who received unilateral CI because of severe bilateral sensorineural hearing loss were assessed for cVEMP produced by TB stimuli at two frequencies (500 and 1000 Hz) in the airway. VEMPs were recorded when the devices were switched to the on (CI-on) and off (CI-off) positions. At the CI-on position, the surgical side (SS) 500-Hz response rates (15/30) were significantly higher than the SS 1000-Hz response rates (9/30) (p=0.031), while the non-operated control side (CS) 500-Hz response rates (20/30) were higher than the CS 1000-Hz response rates (18/30), but the difference was not significant (p=0.50). At the CI-on position, the SS 500-Hz response rates (15/30) were lower than the CS 500-Hz response rates (20/30), but the difference was not significant (p=0.063). However, the SS 1000-Hz response rates (9/30) were significantly lower than the CS 1000-Hz response rates (18/30) (p=0.004). When there was no significant difference between the 500-Hz amplitudes on either side, the SS 1000-Hz amplitudes were found to be significantly lower (p=0.04). CIs have the potential to cause mechanical damage and electrical stimulation to the vestibular system. Possible implant-mediated mechanical damage and electrical stimulation in the high-frequency region affecting the cVEMP response could be found by a 1000-Hz stimulus.


Coffee drinking and cutaneous melanoma risk in the NIH-AARP diet and health study.
Cutaneous melanoma is the fifth most common cancer in the United States. Modifiable risk factors, with the exception of exposure to ultraviolet radiation (UVR), are poorly understood. Coffee contains numerous bioactive compounds and may be associated inversely with melanoma. However, previous epidemiological evidence is limited. Coffee intake was assessed at baseline with a food frequency questionnaire in the National Institutes of Health-AARP prospective cohort study. Among 447 357 non-Hispanic whites who were cancer-free at baseline, 2904 incident cases of malignant melanoma were identified during 4 329 044 person-years of follow-up, with a median of 10.5 years of follow-up. Multivariable-adjusted Cox proportional hazards models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for coffee intake and subsequent melanoma risk with non-coffee drinkers as the reference group. Statistical tests were two-sided, and P values less than .05 were interpreted as statistically significant. The highest category of coffee intake was inversely associated with malignant melanoma (≥4 cups/day: HR = 0.80, 95% CI = 0.68 to 0.93, P trend = .01). This association was statistically significant for caffeinated (≥4 cups/day: HR = 0.75, 95% CI = 0.64 to 0.89, P trend = .01) but not for decaffeinated coffee (P trend = .55). Higher coffee intake was associated with a modest decrease in risk of melanoma in this large US cohort study. Additional investigations of coffee intake and its constituents, particularly caffeine, with melanoma are warranted.


Learning: rapid aversive conditioning in the gastropod mollusk Pleurobranchaea.
Untrained Pleurobranchaea feed voraciously when presented food and withdraw from electrical shocks. We trained experimental animals in ten trials spaced 1 hour apart to withdraw from food alone by electrically shocking them if they fed or were indifferent to food. The greatest increase in the number of learned withdrawal responses occurred within 12 hours after conditioning, and was accompanied by long-lasting increased in the threshold and latency of feeding responses. Control animals, which received food and shock alternately (unpaired) every half hour, showed considerably weaker changes than experimentals. These control responses quickly returned to initial levels after conditioning.


Cardiovascular complications in atypical haemolytic uraemic syndrome.
Haemolytic uraemic syndrome (HUS) is characterized by nonimmune haemolytic anaemia, thrombocytopenia and renal impairment-most incidents in childhood are caused by shiga toxin-producing bacteria. Atypical HUS (aHUS) accounts for 10% of cases and has a poor prognosis. About 60% of patients with aHUS have dysregulation of the alternative complement pathway (complement-mediated aHUS). The kidney is the main target organ, but other organs might also be affected. Cardiac complications occur in 3-10% of patients with complement-mediated aHUS, as a consequence of microangiopathic injury in the coronary microvasculature, and can cause sudden death. Emerging evidence also suggests that either thrombosis or stenosis of the medium and large arteries might complicate disease course, and such disorders occur even after renal function is lost. In this Perspectives article we discuss the impact of cardiovascular involvement in complement-mediated aHUS, the role of acute and chronic complement hyperactivation in such events and the implications for treatment.


Single-day and multiday repeatability of stance analysis results for dogs with hind limb lameness.
To assess single-day and multiday repeatability of weight distribution (stance) data obtained with a commercial analyzer for dogs with naturally occurring hind limb lameness. 46 dogs (15 and 31 for single-day and multiday trials, respectively). For single-day trials, 5 to 10 measurements/trial were collected to determine body weight (BW), weight distribution on each limb, and forelimb and hind limb symmetry indices (SIs). The dog was removed from the room and returned immediately; 5 trials were performed. For multiday trials, measurements were performed in the same manner on 2 sequential days. Data were compared among trials (single-day measurements) and between days (multiday measurements). Repeatability (correlation coefficients and Lin concordance correlation coefficients [LCCCs]) and variability (coefficients of variation [CVs]) were assessed. In single-day trials, BW (r = 0.999), weight distribution on the lame hind limb (r = 0.915) and contralateral hind limb (r = 0.948), and hind limb SI (r = 0.964) were each significantly correlated among trials. In multiday trials, BW results were similar; weight distribution on the lame hind limb and contralateral hind limb and hind limb SI were each less closely but still significantly correlated between days. The LCCCs were highest for BW, weight distribution on the contralateral hind limb, and hind limb SI in single-day trials and for BW and weight bearing on the contralateral and lame hind limbs in multiday trials. The CVs were lowest for BW and highest for forelimb SI in both trial types. The analyzer used allowed repeatable measurement of BW and weight distribution on the hind limbs of dogs with hind limb lameness. Measurement of forelimb stance variables was not repeatable in this group of dogs.


Developmental Loci Harbor Clusters of Accelerated Regions That Evolved Independently in Ape Lineages.
Some of the fastest evolving regions of the human genome are conserved noncoding elements with many human-specific DNA substitutions. These human accelerated regions (HARs) are enriched nearby regulatory genes, and several HARs function as developmental enhancers. To investigate if this evolutionary signature is unique to humans, we quantified evidence of accelerated substitutions in conserved genomic elements across multiple lineages and applied this approach simultaneously to the genomes of five apes: human, chimpanzee, gorilla, orangutan, and gibbon. We find roughly similar numbers and genomic distributions of lineage-specific accelerated regions (linARs) in all five apes. In particular, apes share an enrichment of linARs in regulatory DNA nearby genes involved in development, especially transcription factors and other regulators. Many developmental loci harbor clusters of nonoverlapping linARs from multiple apes, suggesting that accelerated evolution in each species affected distinct regulatory elements that control a shared set of developmental pathways. Our statistical tests distinguish between GC-biased and unbiased accelerated substitution rates, allowing us to quantify the roles of different evolutionary forces in creating linARs. We find evidence of GC-biased gene conversion in each ape, but unbiased acceleration consistent with positive selection or loss of constraint is more common in all five lineages. It therefore appears that similar evolutionary processes created independent accelerated regions in the genomes of different apes, and that these lineage-specific changes to conserved noncoding sequences may have differentially altered expression of a core set of developmental genes across ape evolution.


Immunohistochemical localization of the immunodominant differentiation antigen lacto-N-fucopentaose III in normal adult and fetal tissues.
Monoclonal antibodies were produced by immunizing rats with human small cell lung carcinoma (SCLC) cell lines. Monoclonal antibodies 600D11 and 624A12 were found to be directed against the ceramide pentasaccharide that contains the lacto-N-fucopentaose III (LNFP III) sequence of sugars, an isomer of the Lewis A blood group antigen. LNFP III is an immunodominant antigen whose reactivity is maintained in formalin-fixed paraffin-embedded sections (PS). LNFP III has been recognized in a number of human tumors including: SCLC; adenocarcinomas of the breast, gastrointestinal tract, genitourinary tract, and lung; renal cell carcinoma; neuroblastoma; and myelogenous leukemia. We now report the normal adult and fetal tissue distribution of the LNFP III antigen by immunoperoxidase staining on PS utilizing 600D11 and 624A12. Binding was demonstrated in bronchial epithelium and bronchial glands; squamous epithelium of the esophagus; gastric crypts, duodenal enterocytes and Brunners glands; argentaffin cells; jejunal and colonic goblet cells; pancreatic acinar cells; salivary glands; endocervical and exocervical cells; skin epidermis; myelinated motor fibers; cells of the adrenal medulla and anterior pituitary gland; polymorphonuclear leukocytes (PMNs); tissue macrophages and renal proximal tubules and loops of Henle. Staining was localized to cell membranes and within the cytoplasm, with greatest intensity at the apical and basal portions of the cells. These staining patterns were noted in adult and neonatal tissues, and initial expression could be traced to approximately the second trimester of fetal development. Knowledge of the normal tissue distribution of this immunodominant antigenic determinant may offer insight into its structural and functional role in benign and malignant tissues.


Ion Channels Made from a Single Membrane-Spanning DNA Duplex.
Because of their hollow interior, transmembrane channels are capable of opening up pathways for ions across lipid membranes of living cells. Here, we demonstrate ion conduction induced by a single DNA duplex that lacks a hollow central channel. Decorated with six porpyrin-tags, our duplex is designed to span lipid membranes. Combining electrophysiology measurements with all-atom molecular dynamics simulations, we elucidate the microscopic conductance pathway. Ions flow at the DNA-lipid interface as the lipid head groups tilt toward the amphiphilic duplex forming a toroidal pore filled with water and ions. Ionic current traces produced by the DNA-lipid channel show well-defined insertion steps, closures, and gating similar to those observed for traditional protein channels or synthetic pores. Ionic conductances obtained through simulations and experiments are in excellent quantitative agreement. The conductance mechanism realized here with the smallest possible DNA-based ion channel offers a route to design a new class of synthetic ion channels with maximum simplicity.


Plagiarism in residency application essays.
Anecdotal reports suggest that some residency application essays contain plagiarized content. To determine the prevalence of plagiarism in a large cohort of residency application essays. Retrospective cohort study. 4975 application essays submitted to residency programs at a single large academic medical center between 1 September 2005 and 22 March 2007. Specialized software was used to compare residency application essays with a database of Internet pages, published works, and previously submitted essays and the percentage of the submission matching another source was calculated. A match of more than 10% to an existing work was defined as evidence of plagiarism. Evidence of plagiarism was found in 5.2% (95% CI, 4.6% to 5.9%) of essays. The essays of non-U.S. citizens were more likely to demonstrate evidence of plagiarism. Other characteristics associated with the prevalence of plagiarism included medical school location outside the United States and Canada; previous residency or fellowship; lack of research experience, volunteer experience, or publications; a low United States Medical Licensing Examination Step 1 score; and non-membership in the Alpha Omega Alpha Honor Medical Society. The software database is probably incomplete, the 10%-match threshold for defining plagiarism has not been statistically validated, and the study was confined to applicants to 1 institution. Evidence of matching content in an essay cannot be used to infer the applicant's intent and is not sensitive to variations in the cultural context of copying in some societies. Evidence of plagiarism in residency application essays is more common in international applicants but was found in those by applicants to all specialty programs, from all medical school types, and even among applicants with significant academic honors. No external funding.


CD133 antisense suppresses cancer cell growth and increases sensitivity to cisplatin in vitro.
The increased incidence of cancer in recent years is associated with a high rate of mortality. Numerous types of cancer have a low percentage of CD133(+) cells, which have similar features to stem cells. The CD133 molecule is involved in apoptosis and cell proliferation. The aim of this study was to determine the biological effect of CD133 suppression and its role in the chemosensitization of cancer cell lines. RT-PCR and immunocytochemical analyses indicated that CD133 was expressed in the cancer cell lines B16F10, MCF7 and INER51. Downregulation of CD133 by transfection with an antisense sequence (As-CD133) resulted in a decrease in cancer cell viability of up to 52, 47 and 22% in B16F10, MCF-7 and INER51 cancer cell lines, respectively. This decreased viability appeared to be due to the induction of apoptosis. In addition, treatment with As-CD133 in combination with cisplatin had a synergic effect in all of the cancer cell lines analyzed, and in particular, significantly decreased the viability of B16F10 cancer cells compared with each treatment separately (3.1% viability for the combined treatment compared with 48% for 0.4 μg As-CD133 and 25% for 5 ng/μl cisplatin; P<0.05). The results indicate that the downregulation of CD133 by antisense is a potential therapeutic target for cancer and has a synergistic effect when administered with minimal doses of the chemotherapeutic drug cisplatin, suggesting that this combination strategy may be applied in cancer treatment.


The prophylactic use of probiotics in the prevention of radiation therapy-induced diarrhea.
Cancer treatment regimens that include radiation therapy (RT) to the abdominal region for cervical, ovarian, prostate, sigmoid, or colorectal cancer potentially disturb the colonization resistance of the indigenous gut flora, causing RT-induced diarrhea, enteritis, and colitis in more than 80% of patients with cancer. One approach for the prevention of RT-induced diarrhea is the use of probiotics. Randomized clinical trials have demonstrated efficacy of probiotic preparations VSL #3 and Lactobacillus casei DN-114 001 in decreasing the incidence and grade of RT-induced diarrhea. Oncology nurses and advanced practice clinicians are in a position to interpret research findings related to RT-induced diarrhea, enteritis, and colitis and to apply evidence-based practice principles in patients with cancer receiving RT to promote positive outcomes.


Localization of association signal from risk and protective variants in sequencing studies.
Aggregating information across multiple variants in a gene or region can improve power for rare variant association testing. Power is maximized when the aggregation region contains many causal variants and few neutral variants. In this paper, we present a method for the localization of the association signal in a region using a sliding-window based approach to rare variant association testing in a region. We first introduce a novel method for analysis of rare variants, the Difference in Minor Allele Frequency test (DMAF), which allows combined analysis of common and rare variants, and makes no assumptions about the direction of effects. In whole-region analyses of simulated data with risk and protective variants, DMAF and other methods which pool data across individuals were found to outperform methods which pool data across variants. We then implement a sliding-window version of DMAF, using a step-down permutation approach to control type I error with the testing of multiple windows. In simulations, the sliding-window DMAF improved power to detect a causal sub-region, compared to applying DMAF to the whole region. Sliding-window DMAF was also effective in localizing the causal sub-region. We also applied the DMAF sliding-window approach to test for an association between response to the drug gemcitabine and variants in the gene FKBP5 sequenced in 91 lymphoblastoid cell lines derived from white non-Hispanic individuals. The application of the sliding-window test procedure detected an association in a sub-region spanning an exon and two introns, when rare and common variants were analyzed together.


The feasibility of measuring bone uranium concentrations in vivo using source excited K x-ray fluorescence.
X-ray fluorescence (XRF) has been demonstrated to be an extremely useful technique for measuring trace quantities of heavy metals in various tissues within the body. This study investigates the applicability of XRF to the measurement of bone uranium concentration. The factors affecting the accuracy and minimal detectable concentration of bone uranium in vivo are discussed. The system chosen employs a 57Co source to excite the uranium K x-rays, with the source and detector in an approximate 180 degrees backscatter geometry relative to the sample position. It is demonstrated, with experiment and Monte Carlo simulation, that the x-ray to coherent peak ratio is linearly related to concentration and is independent of variations in source-sample geometry, thickness of overlying tissue and tibia size. Preliminary in vivo measurements indicate that this system has a minimum detectable concentration of approximately 20 micrograms U/g bone mineral which may not be sufficiently precise for a monitoring system for occupational workers. Future work will involve attempts to enhance this precision through the use of alternative sources for the excitation of uranium x-rays as well as possible modifications to the detector and electronics.


Inhibition of Shikimate Kinase and Type II Dehydroquinase for Antibiotic Discovery: Structure-Based Design and Simulation Studies.
The loss of effectiveness of current antibiotics caused by the development of drug resistance has become a severe threat to public health. Current widely used antibiotics are surprisingly targeted at a few bacterial functions - cell wall, DNA, RNA, and protein biosynthesis - and resistance to them is widespread and well identified. There is therefore great interest in the discovery of novel drugs and therapies to tackle antimicrobial resistance, in particular drugs that target other essential processes for bacterial survival. In the past few years a great deal of effort has been focused on the discovery of new inhibitors of the enzymes involved in the biosynthesis of aromatic amino acids, also known as the shikimic acid pathway, in which chorismic acid is synthesized. The latter compound is the synthetic precursor of L-Phe, L-Tyr, L-Phe, and other important aromatic metabolites. These enzymes are recognized as attractive targets for the development of new antibacterial agents because they are essential in important pathogenic bacteria, such as Mycobacterium tuberculosis and Helicobacter pylori, but do not have any counterpart in human cells. This review is focused on two key enzymes of this pathway, shikimate kinase and type II dehydroquinase. An overview of the use of structure-based design and computational studies for the discovery of selective inhibitors of these enzymes will be provided. A detailed view of the structural changes caused by these inhibitors in the catalytic arrangement of these enzymes, which are responsible for the inhibition of their activity, is described.


Evidence-based pediatric pain management in emergency departments of a rural state.
Evidence-based pediatric pain management (EBPPM) has been identified as a practice too often overlooked in Emergency Departments (EDs). Studies show EBPPM is practiced inconsistently in urban EDs, and even less is known about the practice in rural EDs. The objectives of this study were: A) Determine the frequencies with which specific EBPPM practices are used in EDs of a primarily rural state; and B) Explore the differences in EBPPM practice in Critical Access, rural, and urban hospital EDs. A web-based survey, measuring the use of 14 EBPPM practices, was offered to all licensed independent providers (Medical Doctors, Doctors of Osteopathy, Physicians' Assistants, and Advanced Registered Nurse Practitioners) and nurses from the 118 hospital EDs in a rural state. Responses from 259 providers and 1,177 nurses revealed that the majority of respondents infrequently used any type of topical analgesic before venipuncture or IV insertion in children, or oral sucrose for infant procedures. Tests for group differences show that providers from urban EDs more frequently used a topical analgesic for suturing lacerations, provided analgesics for blood draws, and gave pain medication to children with abdominal pain. Nurses from urban hospitals used significantly more EBPPM practices than nurses from Critical Access and rural hospitals (P < .001). In hospitals of all types, ED providers and nurses fail to take advantage of EBPPM practices. This study reveals that health professionals in rural settings are particularly in need of improving the use of recommended pediatric pain management practices.


Bioactive arabinogalactans from the leaves of Opilia celtidifolia Endl. ex Walp. (Opiliaceae).
The leaves of the tree Opilia celtidifolia have a long tradition for being used in Mali and other West African countries against various ailments such as for wound healing. Previous studies on polysaccharides from these leaves showed the presence of pectic-like polymers with an effect on the human complement system as well as the ability to activate macrophages. The present study shows that bioactive arabinogalactans isolated by water of 50°C could be separated into two acidic fractions, Oc50A1 and Oc50A2. The former could, by gel filtration on Sephacryl S-400, be separated into two fractions, which were further purified on a Superdex 200 column to give the fractions Oc50A1.I.pur and Oc50A1.II.pur. These fractions were subjected to chemical and biological studies. The polysaccharides consisted mainly of heavily branched type II arabinogalactans and minor amounts of rhamnogalacturonan I regions. The isolated polymers had a high human complement-fixating ability, as well as the ability to stimulate rat macrophages and dendritic cells (DCs) and to induce B cell proliferation. These effects were especially pronounced for the higher molecular weight fraction of Oc50A1.I.pur. The fractions Oc50A1.I.pur and Oc501.II.pur stimulated secretion of pro-inflammatory cytokines from purified B cells or DCs. Collectively, these results indicate that the arabinogalactan type II polymers present in the leaves of O. celtidifolia may be used to develop medical devices for regulating inflammatory processes.


Effect of Nitrogen Sources on Omega-3 Polyunsaturated Fatty Acid Biosynthesis and Gene Expression in Thraustochytriidae sp.
The molecular mechanism that contributes to nitrogen source dependent omega-3 polyunsaturated fatty acid (n-3 PUFA) synthesis in marine oleaginous protists Thraustochytriidae sp., was explored in this study. The fatty acid (FA) synthesis was significantly influenced by the supplement of various levels of sodium nitrate (SN) (1-50 mM) or urea (1-50 mM). Compared with SN (50 mM) cultivation, cells from urea (50 mM) cultivation accumulated 1.16-fold more n-3 PUFAs (49.49% docosahexaenoic acid (DHA) (w/w, of total FAs) and 5.28% docosapentaenoic acid (DPA) (w/w, of total FAs)). Strikingly higher quantities of short chain FAs (<18 carbons) (52.22-fold of that in urea cultivation) were produced from SN cultivation. Ten candidate reference genes (RGs) were screened by using four statistical methods (geNorm, NormFinder, Bestkeeper and RefFinder). MFT (Mitochondrial folate transporter) and NUC (Nucleolin) were determined as the stable RGs to normalize the RT-qPCR (real-time quantitative polymerase chain reaction) data of essential genes related to n-3 PUFAs-synthesis. Our results elucidated that the gene transcripts of delta(3,5)-delta(2,4)-dienoyl-CoA isomerase, enoyl-CoA hydratase, fatty acid elongase 3, long-chain fatty acid acyl-CoA ligase, and acetyl-CoA carboxylase were up-regulated under urea cultivation, contributing to the extension and unsaturated bond formation. These findings indicated that regulation of the specific genes through nitrogen source could greatly stimulate n-3 PUFA production in Thraustochytriidae sp.


An analysis of 1539 patients with cancer of unknown primary site.
A retrospective review of 1539 patients with cancer of unknown primary site seen at Yale-New Haven Hospital from 1922 to 1981 was performed. Information was obtained from the Tumor Registry. The method of diagnosis, patient characteristics, year of diagnosis, histologic features, treatment received, and survival were analyzed. The most common cell type was adenocarcinoma. Survival overall was poor, with a median survival of 5 months for the entire group. Age and year of diagnosis did not appear to significantly influence survival. Closer examination of a small subset of those patients with squamous cell carcinoma revealed a very high male:female ratio, possibly related to tobacco and alcohol abuse. Nearly 9% of these patients were found to have a history of one or more unrelated malignancies.


A 40-million-year lake record of early mesozoic orbital climatic forcing.
Sediments of the early Mesozoic Newark Supergroup of eastern North America consist largely of sedimentary cycles produced by the rise and fall of very large lakes that responded to periodic climate changes controlled by variations in the earth's orbit. Fourier analysis of long sections of the Late Triassic Lockatong and Passaic formations of the Newark Basin show periods in thickness of 5.9, 10.5, 25.2, 32.0, and 96.0 meters corresponding to periodicities in time of roughly 25,000, 44,000, 100,0003,, 13000 and 400,000 years, as judged by radiometric time scales and varve-calibrated sedimentation rates. The ratios of the shortest cycle with longer cycles correspond closely to the ratios of the present periods of the main orbital terms that appear to influence climate. Similar long sequences of sedimentary cycles occur through most of the rest of the Newark Supergroup spanning a period of more than 40 million years. This is strong evidence of orbital forcing of climate in the ice-free early Mesozoic and indicates that the main periods of the orbital cycles were not very different 200 million years ago from those today.