Escherichia coli contamination of lettuce grown in soils amended with animal slurry.
A pilot study was conducted to assess the transfer of Escherichia coli from animal slurry fertilizer to lettuce, with E. coli serving as an indicator of fecal contamination and as an indicator for potential bacterial enteric pathogens. Animal slurry was applied as fertilizer to three Danish agricultural fields prior to the planting of lettuce seedlings. At harvest, leaves (25 g) of 10 lettuce heads were pooled into one sample unit (n = 147). Soil samples (100 g) were collected from one field before slurry application and four times during the growth period (n = 75). E. coli was enumerated in slurry, soil, and lettuce on 3M Petrifilm Select E. coli Count Plates containing 16 mg/liter streptomycin, 16 mg/liter ampicillin, or no antimicrobial agent. Selected E. coli isolates (n = 83) originating from the slurry, soil, and lettuce were genotyped by pulsed-field gel electrophoresis (PFGE) to determine the similarity of isolates. The slurry applied to the fields contained 3.0 to 4.5 log CFU/g E. coli. E. coli was found in 36 to 54% of the lettuce samples, streptomycin-resistant E. coli was found in 10.0 to 18.0% of the lettuce samples, and ampicillin-resistant E. coli in 0 to 2.0% of the lettuce samples (the detection limit was 1 log CFU/g). The concentration of E. coli exceeded 2 log CFU/g in 19.0% of the lettuce samples. No E. coli was detected in the soil before the slurry was applied, but after, E. coli was present until the last sampling day (harvest), when 10 of 15 soil samples contained E. coli. A relatively higher frequency of E. coli in lettuce compared with the soil samples at harvest suggests environmental sources of fecal contamination, e.g., wildlife. The higher frequency was supported by the finding of 21 different PFGE types among the E. coli isolates, with only a few common PFGE types between slurry, soil, and lettuce. The frequent finding of fecal-contaminated lettuce indicates that human pathogens such as Salmonella and Campylobacter can be present and represent food safety hazards.


Gliolectin positively regulates Notch signalling during wing-vein specification in Drosophila.
Notch signalling is essential for animal development. It integrates multiple pathways controlling cell fate and specification. Here we report the genetic characterization of Gliolectin, presumably a lectin, a cytoplasmic protein, significantly enriched in Golgi bodies. Its expression overlaps with regions where Notch is activated. Loss of gliolectin function results in ectopic veins, while gain of its function causes loss of wing veins. It positively regulates Enhancer of split mβ, a target of Notch signalling. These observations suggest that it is a positive regulator of Notch signalling during wing development in Drosophila.


Nanoscopic spine localization of Norbin, an mGluR5 accessory protein.
Norbin is a neuron-specific, cytosolic protein that interacts with the metabotropic glutamate receptor 5 (mGluR5) and has a profound impact on mGluR5 signaling. Yet, little is known about its synaptic distribution. Here we have analyzed the spatial relationship between Norbin, postsynaptic density protein 95 (PSD-95), actin and mGluR5 in spines using super-resolution microscopy. Norbin was found to have a high degree of colocalization with actin and a lower degree of colocalization with PSD-95. Co-immunoprecipitation studies confirmed that interaction occurs between Norbin and actin, but not between Norbin and PSD-95. Norbin was also found to have a high degree of colocalization with the perisynaptically located mGluR5. Findings based on structured illumination microscopy (3D-SIM) of exogenous expressed Norbin-GFP were confirmed by stimulated emission depletion microscopy (STED) of immunolabeled endogenous Norbin. Norbin associates with actin rather than with PSD-95 in dendritic spines. Results regarding protein localization and colocalization performed with conventional confocal microscopy must be interpreted with great caution. The now available super-resolution microscopy techniques provide more accurate information about sub-cellular protein localization than previously was possible.


Proteome specialization of anaerobic fungi during ruminal degradation of recalcitrant plant fiber.
The rumen harbors a complex microbial mixture of archaea, bacteria, protozoa, and fungi that efficiently breakdown plant biomass and its complex dietary carbohydrates into soluble sugars that can be fermented and subsequently converted into metabolites and nutrients utilized by the host animal. While rumen bacterial populations have been well documented, only a fraction of the rumen eukarya are taxonomically and functionally characterized, despite the recognition that they contribute to the cellulolytic phenotype of the rumen microbiota. To investigate how anaerobic fungi actively engage in digestion of recalcitrant fiber that is resistant to degradation, we resolved genome-centric metaproteome and metatranscriptome datasets generated from switchgrass samples incubated for 48 h in nylon bags within the rumen of cannulated dairy cows. Across a gene catalog covering anaerobic rumen bacteria, fungi and viruses, a significant portion of the detected proteins originated from fungal populations. Intriguingly, the carbohydrate-active enzyme (CAZyme) profile suggested a domain-specific functional specialization, with bacterial populations primarily engaged in the degradation of hemicelluloses, whereas fungi were inferred to target recalcitrant cellulose structures via the detection of a number of endo- and exo-acting enzymes belonging to the glycoside hydrolase (GH) family 5, 6, 8, and 48. Notably, members of the GH48 family were amongst the highest abundant CAZymes and detected representatives from this family also included dockerin domains that are associated with fungal cellulosomes. A eukaryote-selected metatranscriptome further reinforced the contribution of uncultured fungi in the ruminal degradation of recalcitrant fibers. These findings elucidate the intricate networks of in situ recalcitrant fiber deconstruction, and importantly, suggest that the anaerobic rumen fungi contribute a specific set of CAZymes that complement the enzyme repertoire provided by the specialized plant cell wall degrading rumen bacteria.


Protocolled practice nurse-led care for children with asthma in primary care: protocol for a cluster randomised trial.
In children with asthma, daily symptoms and exacerbations have a significant impact on the quality of life of both children and parents. More effective use of asthma medication and, consequently, better asthma control is advocated, since both overtreatment and undertreatment are reported in primary care. Trials in adults suggest that asthma control is better when patients receive a regular medical review. Therefore, protocolled care by the general practitioner may also lead to better asthma control in children. However, such protocolled care by the general practitioner may be time consuming and less feasible. Therefore, this study aims to determine whether protocolled practice nurse-led asthma care for children in primary care provides more effective asthma control than usual care. The study will be a cluster-randomised open-label trial with an 18-month follow-up. Practice nurses will be the units of randomisation and children with asthma the units of analysis. It is planned to include 180 children aged 6-12 years. Primary outcome will be average asthma control during the 18-month follow-up measured by the Childhood Asthma Control Test (C-ACT). Secondary outcomes include C-ACT scores at t=3, t=6, t=12 and t=18 months; the frequency and severity of exacerbations; cost-effectiveness; quality of life; satisfaction with delivered care; forced expiratory volume in 1 s and forced expiratory flow at 75% and the association of high symptoms scores at baseline and baseline characteristics. Besides, we will conduct identical measurements in a non-randomised sample of children. This will be the first trial to evaluate the effectiveness of protocolled practice nurse-led care for children with asthma in primary care. The results may lead to improvements in asthma care for children and can be directly implemented in revisions of asthma guidelines.The study protocol was approved by the Medical Research Ethics Committee of the Erasmus Medical Centre in Rotterdam. NTR6847.


Graph theoretical analysis of resting-state MEG data: Identifying interhemispheric connectivity and the default mode.
Interhemispheric connectivity with resting state MEG has been elusive, and demonstration of the default mode network (DMN) yet more challenging. Recent seed-based MEG analyses have shown interhemispheric connectivity using power envelope correlations. The purpose of this study is to compare graph theoretic maps of brain connectivity generated using MEG with and without signal leakage correction to evaluate for the presence of interhemispheric connectivity. Eight minutes of resting state eyes-open MEG data were obtained in 22 normal male subjects enrolled in an IRB-approved study (ages 16-18). Data were processed using an in-house automated MEG processing pipeline and projected into standard (MNI) source space at 7mm resolution using a scalar beamformer. Mean beta-band amplitude was sampled at 2.5second epochs from the source space time series. Leakage correction was performed in the time domain of the source space beam formed signal prior to amplitude transformation. Graph theoretic voxel-wise source space correlation connectivity analysis was performed for leakage corrected and uncorrected data. Degree maps were thresholded across subjects for the top 20% of connected nodes to identify hubs. Additional degree maps for sensory, visual, motor, and temporal regions were generated to identify interhemispheric connectivity using laterality indices. Hubs for the uncorrected MEG networks were predominantly symmetric and midline, bearing some resemblance to fMRI networks. These included the cingulate cortex, bilateral inferior frontal lobes, bilateral hippocampal formations and bilateral cerebellar hemispheres. These uncorrected networks however, demonstrated little to no interhemispheric connectivity using the ROI-based degree maps. Leakage corrected MEG data identified the DMN, with hubs in the posterior cingulate and biparietal areas. These corrected networks demonstrated robust interhemispheric connectivity for the ROI-based degree maps. Graph theoretic analysis of MEG resting state data without signal leakage correction can demonstrate symmetric networks with some resemblance to fMRI networks. These networks however, are an artifact of high local correlation from signal leakage and lack interhemispheric connectivity. Following signal leakage correction, MEG hubs emerge in the DMN, with strong interhemispheric connectivity.


Structure-based discovery of NANOG variant with enhanced properties to promote self-renewal and reprogramming of pluripotent stem cells.
NANOG (from Irish mythology Tír na nÓg) transcription factor plays a central role in maintaining pluripotency, cooperating with OCT4 (also known as POU5F1 or OCT3/4), SOX2, and other pluripotency factors. Although the physiological roles of the NANOG protein have been extensively explored, biochemical and biophysical properties in relation to its structural analysis are poorly understood. Here we determined the crystal structure of the human NANOG homeodomain (hNANOG HD) bound to an OCT4 promoter DNA, which revealed amino acid residues involved in DNA recognition that are likely to be functionally important. We generated a series of hNANOG HD alanine substitution mutants based on the protein-DNA interaction and evolutionary conservation and determined their biological activities. Some mutant proteins were less stable, resulting in loss or decreased affinity for DNA binding. Overexpression of the orthologous mouse NANOG (mNANOG) mutants failed to maintain self-renewal of mouse embryonic stem cells without leukemia inhibitory factor. These results suggest that these residues are critical for NANOG transcriptional activity. Interestingly, one mutant, hNANOG L122A, conversely enhanced protein stability and DNA-binding affinity. The mNANOG L122A, when overexpressed in mouse embryonic stem cells, maintained their expression of self-renewal markers even when retinoic acid was added to forcibly drive differentiation. When overexpressed in epiblast stem cells or human induced pluripotent stem cells, the L122A mutants enhanced reprogramming into ground-state pluripotency. These findings demonstrate that structural and biophysical information on key transcriptional factors provides insights into the manipulation of stem cell behaviors and a framework for rational protein engineering.


Secreted microvesicular miR-31 inhibits osteogenic differentiation of mesenchymal stem cells.
Damage to cells and tissues is one of the driving forces of aging and age-related diseases. Various repair systems are in place to counteract this functional decline. In particular, the property of adult stem cells to self-renew and differentiate is essential for tissue homeostasis and regeneration. However, their functionality declines with age (Rando, 2006). One organ that is notably affected by the reduced differentiation capacity of stem cells with age is the skeleton. Here, we found that circulating microvesicles impact on the osteogenic differentiation capacity of mesenchymal stem cells in a donor-age-dependent way. While searching for factors mediating the inhibitory effect of elderly derived microvesicles on osteogenesis, we identified miR-31 as a crucial component. We demonstrated that miR-31 is present at elevated levels in the plasma of elderly and of osteoporosis patients. As a potential source of its secretion, we identified senescent endothelial cells, which are known to increase during aging in vivo (Erusalimsky, 2009). Endothelial miR-31 is secreted within senescent cell-derived microvesicles and taken up by mesenchymal stem cells where it inhibits osteogenic differentiation by knocking down its target Frizzled-3. Therefore, we suggest that microvesicular miR-31 in the plasma of elderly might play a role in the pathogenesis of age-related impaired bone formation and that miR-31 might be a valuable plasma-based biomarker for aging and for a systemic environment that does not favor cell-based therapies whenever osteogenesis is a limiting factor.


The Dynamics of Implicit Intergroup Biases of Victims and Ex-combatants in Post-conflict Scenarios.
Strong group identities arise in intergroup conflict scenarios and perpetuate sectarian violence even in post-conflict scenarios. In particular, out-group negative implicit associations are predictors of decreased intergroup forgiveness, as well as increased distrust and aggression against the out-group. Thus, the presence of implicit intergroup (i.e., ex-combatants and victims) biases seems to be a relevant factor in post-conflict scenarios. Here, we aimed to explore whether negative biases toward the out-group are boosted by (a) previous exposure to conflict violence or (b) identification with an armed violent group. One hundred and twenty-eight participants, 65 ex-combatants from Colombian guerrillas and 63 victims of the armed conflict, were assessed with a modified version of the implicit association test (IAT). Our results revealed that the victim group showed a significant negative bias against ex-combatants. However, no bias toward the out-group (i.e., victims) or in-group favoritism was observed in the ex-combatant group. Similarly, we found that IAT scores were not associated with sociodemographical variables (i.e., sex, years of education, or type of dwelling), the levels of combat exposure, victimization armed-conflict-related experiences, or child abuse antecedents. Our results showed an unexpected lack of in-group bias in ex-combatants, potentially triggered by the effect of current demobilization and reintegration processes. Thus, negative associations with the out-group will persist in the framework of societal condemnation of the out-group. In contrast, these negative biases will tend to be abolished when entering in conflict with larger societal reintegration processes. The results reinforce the idea that reintegration may benefit from interventions at the societal level, including all actors of the conflict. In addition, our findings highlight the importance of implementing victim interventions aimed at reducing stigma and revengeful actions in spaces of collective disarmament.


[Growth secular trend during early childhood in Northern Spain].
To check whether secular changes exist in growth pattern of caucasic children from the North of Spain up to 2 years of age compared with 25-30 years ago population growth curves. Longitudinal study of a representative cohort of caucasic infants born a term in Aragon (n=1.099). Standard deviation scores (z-scores) for weight, length and body mass index (BMI) have been calculated by two available 25-30 years ago population growth curves (Sobradillo et al. and Ferrández et al.). At all ages, weight, length and BMI mean z-scores significantly vary depending on the growth standard curve used (p≤0.001). At birth, weight z-score is -0.36 (95%CI:-0.42;-0.30) according to Sobradillo et al. and -0.26 (95%CI:-0.29;0.33) according to Ferrández et al. These differences decreased progressively becoming minimum at 2 years (mean weight z-score -0.08 and -0.05; respectively). At this age, length mean z-score is +0.54 (95%CI:0.48;0.61) according to Sobradillo et al. and +0.19 (95%CI:0.12;0.25) according to Ferrández et al. Mean BMI z-scores of our sample showed lower values than selected standards. Current birth weight of caucasic infants from Northern Spain is lower than in the previous generation. At 2 years of age there is a trend of increasing length, with a similar weight, resulting in lower BMI values.


Pathologic classification and clinical behavior of the spectrum of goblet cell carcinoid tumors of the appendix.
Appendiceal tumors exhibiting both neuroendocrine and glandular differentiation are uncommon and have caused difficulty in pathologic classification, prediction of prognosis, and clinical management. Previously, such lesions have been variously designated as adenocarcinoid, goblet cell carcinoid (GCC), and mixed adenocarcinoma carcinoid. In this study, we undertook a retrospective investigation of 63 such cases and classified them as typical GCC (group A) and adenocarcinoma ex GCC on the basis of the histologic features of the tumor at the primary site. The adenocarcinoma ex GCC group was further divided into signet ring cell type (group B) and poorly differentiated adenocarcinoma type (group C). The clinical characteristics and prognosis were compared within these groups and with conventional de novo appendiceal adenocarcinomas. Both groups A and B tumors shared a similar immunoprofile, which included generally focal immunoreactivity for neuroendocrine markers, and a normal intestinal type mucin glycoprotein profile (negative MUC1 expression and preserved MUC2 immunoreactivity). The proliferative index was relatively low in these tumors and slightly increased from groups A to B tumors (11% to 16%). Both beta-catenin and E-cadherin exhibited a normal membranous staining pattern in groups A and B tumors. The poorly differentiated adenocarcinomas ex GCC (group C) demonstrated abnormal p53 and beta-catenin immunoreactivity. The mean follow-up time was 49+/-5 (SE) months. The overall disease-specific survival for all subtypes was 77%, with 46% of patients without evidence of disease and 31% alive with disease. The mean survival was 43+/-7 months. All the patients with clinical stage of I or IIA disease had a favorable outcome after appropriate surgery with or without chemotherapy. Although most patients (63%) with GCC presented at an advanced clinical stage, their clinical outcome could be differentiated by subclassification of tumors. The stage IV-matched 5-year survival was 100%, 38%, and 0% for groups A, B, and C, respectively. In conclusion, GCC is a distinctive appendiceal neoplasm that exhibits unique pathologic features and clinical behavior. They display a spectrum of histologic features and possess the potential to transform to an adenocarcinoma phenotype of either signet ring cell or poorly differentiated adenocarcinoma types. Careful evaluation of the morphologic features of GCCs and appropriate pathologic classification are crucial for clinical management and prediction of outcome. Surgical management with right hemicolectomy is recommended after appendectomy for most cases, particularly those with an adenocarcinoma component (groups B and C).


Retroperitoneal Biloma due to Spontaneous Perforation of the Left Hepatic Duct.
Spontaneous perforation of the bile duct in adults is very rare, particularly in cases accompanied by retroperitoneal biloma. We report a patient with retroperitoneal biloma due to a spontaneous perforation of the left hepatic duct. An 82-year-old man was admitted to our institution with abdominal pain and a high fever. He had tenderness at the epi-mesogastrium. Computed tomography showed several stones in the gall bladder and common bile duct (CBD) and a few ascites. A substantial amount of fluid had collected from the dorsal stratum of the duodenum and pancreas head to the right paracolic gutter and anterior side of the right iliopsoas. Laboratory examination revealed a high inflammation score. He underwent emergent laparotomy. Biliary fluid was revealed after the mobilization of the pancreas head, duodenum, and right side of the colon. Bile duct perforation was suspected. Therefore, we exfoliated the dorsal side of the CBD to the cranial side, and intraoperative cholangiography was performed. However, the perforation site could not be detected. Cholecystectomy and choledocholithotomy were performed. A retrograde transhepatic biliary drainage tube was inserted, and primary closure of the CBD incision site was achieved. Postoperative cholangiography revealed leakage from the left hepatic duct near the caudate branch. There are a few reports of spontaneous bile duct perforation cases in the literature, particularly on infants or children with congenital anomalies, but it is rare in adults. It usually causes bile peritonitis, although bile duct perforation should be considered in the differential diagnosis of spontaneous retroperitoneal fluid collection in adults.


Value of cardiac troponin I cutoff concentrations below the 99th percentile for clinical decision-making.
The aim of this study was to evaluate factors influencing the 99th percentile for cardiac troponin I (cTnI) when this cutoff value is established on a highly sensitive assay, and to compare the value of this cutoff to that of lower cutoffs in the prognostic assessment of patients with coronary artery disease. We used the recently refined Access AccuTnI assay (Beckman-Coulter) to assess the distribution of cTnI results in a community population of elderly individuals [PIVUS (Prospective Study of the Vasculature in Uppsala Seniors) study; n = 1005]. The utility of predefined cTnI cutoffs for risk stratification was then evaluated in 952 patients from the FRISC II (FRagmin and Fast Revascularization during InStability in Coronary artery disease) study at 6 months after these patients had suffered acute coronary syndrome. Selection of assay results from a subcohort of PIVUS participants without cardiovascular disease resulted in a decrease of the 99th percentile from 0.044 microg/L to 0.028 microg/L. Men had higher rates of cTnI elevation with respect to the tested thresholds. Whereas the 99th percentile cutoff was not found to be a useful prognostic indicator for 5-year mortality, both the 90th percentile (hazard ratio 3.1; 95% CI 1.9-5.1) and the 75th percentile (hazard ratio 2.8; 95% CI 1.7-4.7) provided useful prognostic information. Sex-specific cutoffs did not improve risk prediction. The 99th percentile of cTnI depends highly on the characteristics of the reference population from which it is determined. This dependence on the reference population may affect the appropriateness of clinical conclusions based on this threshold. However, cTnI cutoffs below the 99th percentile seem to provide better prognostic discrimination in stabilized acute coronary syndrome patients and therefore may be preferable for risk stratification.


What do public housing residents say about their health?
Residents of public housing have poorer health indicators than comparably resourced individuals from the larger community. To identify major health concerns, issues, and barriers to health of community members living in public housing developments, especially as related to cardiovascular disease prevention. To identify similarities and differences between data collected using two methods to inform future health promotion programs and policies. Key informant interviews were conducted with resident leaders and analyzed qualitatively in eight housing developments. Results were compared with quantitative data collected from a resident health survey with a large sample that analyzed individual and development-level characteristics, major health concerns, and barriers. Several development-level characteristics were significantly associated with residents' health concerns and barriers, including development size, percentage of Spanish speakers, and presence of a tenant task force (TTF); important health promotion barriers included lack of resident engagement, inconsistency in programming, lack of knowledge of actions to prevent chronic disease, and lack of resources for health promotion. Safety-related health concerns were named as a priority. Multiple data collection methods can yield important data about community health priorities and barriers; areas of difference and similarity between methods are especially useful in guiding health promotion efforts and opportunities.


TranscriptClean: variant-aware correction of indels, mismatches and splice junctions in long-read transcripts.
Long-read, single-molecule sequencing platforms hold great potential for isoform discovery and characterization of multi-exon transcripts. However, their high error rates are an obstacle to distinguishing novel transcript isoforms from sequencing artifacts. Therefore, we developed the package TranscriptClean to correct mismatches, microindels and noncanonical splice junctions in mapped transcripts using the reference genome while preserving known variants. Our method corrects nearly all mismatches and indels present in a publically available human PacBio Iso-seq dataset, and rescues 39% of noncanonical splice junctions. All Python and R scripts used in this paper are available at https://github.com/dewyman/TranscriptClean.


Local Lung Mass Masquerading a Very Aggressive Extraskeletal Ewing Sarcoma Presenting as Bilateral Paraparesis in a Young Adult.
Ewing sarcoma is typically seen in children involving long bones. Although well described, its presentation in extraskeletal tissues is relatively rare and is classified as an Ewing sarcoma family of tumors. They are mostly curable when they occur in children. An extraskeletal Ewing sarcoma in adults is uncommon, limiting the experience in adult oncologists. The biopsy is essential for definitive diagnosis, which shows small round blue cells that must be differentiated from lymphoma, embryonal rhabdomyosarcoma, and small cell carcinoma. Management is multimodal, involving surgery, radiation for local treatment of primary tumor, and systemic chemotherapy. A multidisciplinary approach, coupled with risk-adapted intensive neoadjuvant and adjuvant multi-agent chemotherapies and other modalities such as radiation and surgery for control of the primary site and metastatic disease, is needed. The primary multidrug chemotherapy regimen consists of alternating cycles of vincristine/doxorubicin/cyclophosphamide (VDC) and ifosfamide/etoposide (IE) given every 2 weeks with growth factor support. Prognosis and the 5-year survival rate are better for localized than the metastatic disease, and in metastatic disease, it is better for patients with lung metastasis than other metastatic disease sites. We describe a rare extraskeletal tumor arising from a lung that tested positive for Ewing sarcoma, also known as Askin's tumor in a young adult. In our case, the tumor rapidly metastasized locally to involve the thoracic spine causing paraparesis. Timely diagnosis and early management are essential to improve outcomes. We also present how treatment can be delayed due to sepsis and emphasize the careful multispecialty approach's importance.


HuR is exported to the cytoplasm in oral cancer cells in a different manner from that of normal cells.
HuR, a ubiquitously expressed member of the Hu protein family that binds and stabilizes an AU-rich element (ARE)-containing mRNAs, is known to shuttle between the nucleus and the cytoplasm via several export pathways. When normal cells were treated with heat shock, HuR was exported to the cytoplasm in a chromosome maintenance region 1 (CRM1)-dependent manner. However, in this study, we demonstrate that HuR is exported to the cytoplasm in oral cancer cells even if the cells were treated with the inhibitor of the CRM1-independent export pathway. Immunohistochemical and biochemical analyses showed that HuR existed in both the cytoplasm and the nucleus in oral cancer cells, such as HSC-3 and Ca9.22, but existed entirely inside the nucleus in normal cells. AU-rich element-mRNAs were also exported to the cytoplasm and stabilised in the oral cancer cells, which were inhibited by HuR knockdown. This export of HuR was not affected by at least 7 h of treatment of leptomycin B (LMB), which is an inhibitor of the CRM1-dependent export pathway. These findings suggest that HuR is exported to the cytoplasm in oral carcinoma cells in a different manner from that of normal cells, and is likely to occur through the perturbation of a normal export pathway.


Cheating does not explain selective differences at high and low relatedness in a social amoeba.
Altruism can be favored by high relatedness among interactants. We tested the effect of relatedness in experimental populations of the social amoeba Dictyostelium discoideum, where altruism occurs in a starvation-induced social stage when some amoebae die to form a stalk that lifts the fertile spores above the soil facilitating dispersal. The single cells that aggregate during the social stage can be genetically diverse, which can lead to conflict over spore and stalk allocation. We mixed eight genetically distinct wild isolates and maintained twelve replicated populations at a high and a low relatedness treatment. After one and ten social generations we assessed the strain composition of the populations. We expected that some strains would be out-competed in both treatments. In addition, we expected that low relatedness might allow the persistence of social cheaters as it provides opportunity to exploit other strains. We found that at high relatedness a single clone prevailed in all twelve populations. At low relatedness three clones predominated in all twelve populations. Interestingly, exploitation of some clones by others in the social stage did not explain the results. When we mixed each winner against the pool of five losers, the winner did not prevail in the spores because all contributed fairly to the stalk and spores. Furthermore, the dominant clone at high-relatedness was not cheated by the other two that persisted at low relatedness. A combination of high spore production and short unicellular stage most successfully explained the three successful clones at low relatedness, but not why one of them fared better at high relatedness. Differences in density did not account for the results, as the clones did not differ in vegetative growth rates nor did they change the growth rates over relevant densities. These results suggest that social competition and something beyond solitary growth differences occurs during the vegetative stage when amoebae eat bacteria and divide by binary fission. The high degree of repeatability of our results indicates that these effects are strong and points to the importance of new approaches to studying interactions in D. discoideum.


Cardiovascular risk with and without antihypertensive drug treatment in the Japanese general population: participant-level meta-analysis.
To evaluate the cardiovascular mortality risk in association with blood pressure level among people with and without antihypertensive treatment, we performed the participant-level meta-analysis that included 39,705 Japanese from 6 cohorts (58.4% women; mean age, 60.1 years; 20.4% treated). Multivariable-adjusted Cox models were used to analyze the risk of cardiovascular mortality and its subtypes among 6 blood pressure levels according to recent guidelines, optimal to Grade 3 hypertension, and the usage of antihypertensive medication at baseline. During median 10.0 years of follow-up, there were 2032 cardiovascular deaths (5.1 per 1000 person-years), of which 410 deaths were coronary heart disease, 371 were heart failure, and 903 deaths were stroke. Treated participants had significantly higher risk for cardiovascular mortality (hazard ratios, 1.50; 95% confidence intervals, 1.36-1.66), coronary heart disease (hazard ratios, 1.53; confidence intervals, 1.23-1.90), heart failure (hazard ratios, 1.39; confidence intervals, 1.09-1.76), and stroke (hazard ratios, 1.48; confidence intervals, 1.28-1.72) compared with untreated people. Among untreated participants, the risks increased linearly with an increment of blood pressure category (P≤0.011). The risk increments per blood pressure category were higher in young participants (<60 years; 22% to 79%) than those in old people (≥60 years; 7% to 15%) with significant interaction for total cardiovascular, heart failure, and stroke mortality (P≤0.026). Among treated participants, the significant linear association was also observed for cardiovascular mortality (P=0.0003), whereas no stepwise increase in stroke death was observed (P=0.19). The risks of cardiovascular mortality were ≈1.5-fold high in participants under antihypertensive medication. More attention should be paid to the residual cardiovascular risks in treated patients.


National patterns and correlates of mentholated cigarette use in the United States.
To examine the patterns and correlates of mentholated cigarette smoking among adult smokers in the United States. Cross-sectional data on adult current smokers (n = 63,193) were pooled from the 2003 and 2006/07 Tobacco Use Supplements to the Current Population Survey. The associations between socio-demographic and smoking variables were examined with gender- and race/ethnicity-stratified multivariate logistic regression models predicting current use of mentholated cigarettes. Multivariate logistic regression analyses demonstrated that black smokers were 10-11 times more likely to smoke mentholated cigarettes than white smokers men: odds ratio (OR): 11.59, 99% confidence interval (CI): 9.79-13.72; women: OR: 10.12, 99% CI: 8.45-12.11). With the exception of American Indian/Aleut/Eskimo smokers, non-white smokers were significantly more likely to smoke mentholated cigarettes than were white smokers. Additional significant factors associated with mentholated cigarette smoking included being unmarried (never married: OR: 1.21, 99% CI: 1.09-1.34; divorced/separated: OR: 1.13, 99% CI: 1.03-1.23), being born in a US territory (OR: 2.01, 99% CI: 1.35-3.01), living in a non-metropolitan area (OR: 0.87, 99% CI: 0.80-0.96), being unemployed (OR: 1.24, 99% CI: 1.06-1.44) and lower levels of education. Race/ethnicity-stratified analyses showed that women were more likely than men to smoke mentholated cigarettes. Among black smokers, young adults (aged 18-24 years) were four times more likely to smoke mentholated cigarettes compared with individuals aged 65+. Race/ethnicity, gender and age are significant correlates of mentholated cigarette smoking among current smokers. Given the importance of menthol in the cigarette market and the potential untoward health effects of this additive, continued surveillance of the prevalence and correlates of mentholated cigarette use among diverse socio-demographic groups is warranted to inform appropriate interventions.


Dementia in two histologically confirmed cases of multiple sclerosis: one case with isolated dementia and one case associated with psychiatric symptoms.
During the past 10 years, considerable attention has been devoted to cognitive impairment in multiple sclerosis. Occasionally this impairment may be so severe that multiple sclerosis presents as a dementia associated with only minor neurological signs and symptoms. The cases of two women affected by multiple sclerosis who presented with a pure dementia are reported. In the first patient, a progressive apragmatic behavioural disturbance with reduced short term memory and learning abilities were the main clinical features. Neuropathological examination of the brain disclosed numerous plaques in the periventricular white matter, with severe atrophy of the corpus callosum. Plaques were also seen in the white matter of both hippocampus and in the columns of the fornix. The impairment of short term memory could be linked to these lesions. Behavioural changes were probably related to the bilateral lesions of the long associative bundles that disconnected the frontal lobes from other parts of the cerebral hemispheres. In the second patient, visual hallucinations were associated with cognitive dysfunction. MRI showed large plaques in the white matter of both left frontal and temporal lobes. Smaller plaques were also present in the periventricular white matter of the occipital lobes, the nature of which were confirmed by a stereotactic biopsy.


Comparative study of chemical composition and antioxidant activity of fresh and dry rhizomes of turmeric (Curcuma longa Linn.).
The phytoconstituents of essential oil and ethanol oleoresin of fresh and dry rhizomes of turmeric (Curcuma longa Linn.) were analyzed by GC-MS. The major constituents were aromatic-turmerone (24.4%), alpha-turmerone (20.5%) and beta-turmerone (11.1%) in fresh rhizome and aromatic-turmerone (21.4%), alpha-santalene (7.2%) and aromatic-curcumene (6.6%) in dry rhizome oil. Whereas, in oleoresins, the major components were alpha-turmerone (53.4%), beta-turmerone (18.1%) and aromatic-turmerone (6.2%) in fresh and aromatic-turmerone (9.6%), alpha-santalene (7.8%) and alpha-turmerone (6.5%) in dry rhizome. Results showed that alpha-turmerone, a major component in fresh rhizomes is only minor one in dry rhizomes. Also, the content of beta-turmerone in dry rhizomes is less than a half amount found in fresh rhizomes. The antioxidant properties have been assessed by various lipid peroxidation assays as well as DPPH radical scavenging and metal chelating methods. The essential oil and ethanol oleoresin of fresh rhizomes have higher antioxidant properties as compared dry ones.


Complementary use of cross-polarization and standard OCT for differential diagnosis of pathological tissues.
An experimental standard optical coherence tomography (OCT) setup that can be easily modified for cross-polarization OCT (CP OCT) operation has been developed to perform differential diagnosis of pathological tissues. The complementary use of CP OCT, a technique that provides a map of cross-polarization backscattering properties of an object being studied by means of low-coherence interferometry, and standard OCT imaging improves the specificity of diagnostics of pathological changes occurring in tissues. It is shown that healthy, neoplastic and scar tissues of the esophagus have different cross-polarization backscattering properties. A comparative analysis of CP OCT, OCT and histological images from one and the same tissue area has been made. A close correlation between the location of collagen fibers in biological tissue and signal intensity in CP OCT images is found.


Factors contributing to incivility amongst students at a South African nursing school.
This study determined the experiences of nurse educators of the factors contributing to the uncivil classroom behaviours of nursing students at a South African school of nursing. To describe what nurse educators consider to be factors contributing to incivility among nursing students in a South African nursing school. A qualitative descriptive design was used. Eleven nurse educators were purposively sampled for their experiences on the factors contributing to incivility. Individual face-to-face interviews were conducted until data saturation. The data analysed indicated that the educators had varying but often similar perspectives on which factors contribute to incivility among nursing students. The three themes that emerged from the data were academic, psycho-pathological and social factors.The themes were discussed on the basis of their reported impact on classroom behaviour and the implications for the teaching and learning environment. Conclusions were made that an educational screening system to identify committed students before admission into nursing education should be explored; that a support system should be explored for nurse educators to deal with incidents of uncivil behaviour, perhaps within policy frameworks in the nursing institution; that emotional support should be provided for students who may be experiencing difficulties adjusting to the rigours of post-secondary education; and that a forum should be set up for nurse educators to compare notes and share ideas on what works best in reducing the incidence of uncivil behaviours in the classroom setting.


Incidence and predictors of cardiac catheterisation-related arterial thrombosis in children.
Arterial thrombosis is one of the most reported complications of cardiac catheterisation (CC) in children. The aim of the study was to evaluate the incidence and predictors of arterial thrombosis in children with cardiac diseases (CDs). During 12 consecutive months, all children aged 0-19 years undergoing CC of the femoral arteries were included in this observational study. After CC, clinical evaluation of impaired limb perfusion was performed according to local guidelines. Doppler ultrasonography was performed when decreased limb perfusion was suspected. 123 children (30% aged <12 months, 70% aged >12 months) underwent CC. Arterial thrombosis occurred in 14 of the 123 children (11.4%). Twelve cases (12/14=86%) of arterial thrombosis occurred in infants aged <12 months and 2 (2/14=14%) in older children. Overall younger age (p<0.01, OR (95% CI) 0.49 (0.28 to 0.86)) and low body weight (p<0.004, OR (95% CI) 0.78 (0.65 to 0.92)) were significantly associated with an increased risk of arterial thrombosis. Cyanotic CD (p=0.07, OR (95% CI) 2.87 (0.90 to 9.15)) showed a trend towards increased thrombotic risk. Arterial thrombosis is a common complication of CC in infants. Diagnosis of CC-related arterial thrombosis remains a challenge. Well-defined clinical monitoring protocols may be valuable methods for timely detection and treatment of arterial thrombosis.


mBet3p is required for homotypic COPII vesicle tethering in mammalian cells.
TRAPPI is a large complex that mediates the tethering of COPII vesicles to the Golgi (heterotypic tethering) in the yeast Saccharomyces cerevisiae. In mammalian cells, COPII vesicles derived from the transitional endoplasmic reticulum (tER) do not tether directly to the Golgi, instead, they appear to tether to each other (homotypic tethering) to form vesicular tubular clusters (VTCs). We show that mammalian Bet3p (mBet3p), which is the most highly conserved TRAPP subunit, resides on the tER and adjacent VTCs. The inactivation of mBet3p results in the accumulation of cargo in membranes that colocalize with the COPII coat. Furthermore, using an assay that reconstitutes VTC biogenesis in vitro, we demonstrate that mBet3p is required for the tethering and fusion of COPII vesicles to each other. Consistent with the proposal that mBet3p is required for VTC biogenesis, we find that ERGIC-53 (VTC marker) and Golgi architecture are disrupted in siRNA-treated mBet3p-depleted cells. These findings imply that the TRAPPI complex is essential for VTC biogenesis.


Post-traumatic Vertical Gaze Paresis in Nine Patients: Special Vulnerability of the Artery of Percheron in Trauma?
The purpose was to present a case series of vertical gaze paresis in patients with a history of cranioencephalic trauma (CET). The clinical characteristics and management are presented of nine patients with a history of CET secondary to motor vehicle accidents with associated vertical gaze paresis. Neuroimaging studies indicated posttraumatic contusion of the thalamic-mesencephalic region in all nine patients who corresponded to the artery of Percheron region; four patients had signs of hemorrhagic transformation. Vertical gaze paresis was present in all patients, ranging from complete paralysis of the upward and downward gaze to a slight limitation of upward gaze. Posttraumatic vertical gaze paresis is a rare phenomenon that can occur in isolation or in association with other neurological deficits and can cause a significant limitation in the quality-of-life. Studies in the literature have postulated that the unique anatomy of the angle of penetration of the thalamoperforating and lenticulostriate arteries makes these vessels more vulnerable to isolated selective damage in certain individuals and can cause-specific patterns of CET.


Air pollution exposure assessment for epidemiologic studies of pregnant women and children: lessons learned from the Centers for Children's Environmental Health and Disease Prevention Research.
The National Children's Study is considering a wide spectrum of airborne pollutants that are hypothesized to potentially influence pregnancy outcomes, neurodevelopment, asthma, atopy, immune development, obesity, and pubertal development. In this article we summarize six applicable exposure assessment lessons learned from the Centers for Children's Environmental Health and Disease Prevention Research that may enhance the National Children's Study: a) Selecting individual study subjects with a wide range of pollution exposure profiles maximizes spatial-scale exposure contrasts for key pollutants of study interest. b) In studies with large sample sizes, long duration, and diverse outcomes and exposures, exposure assessment efforts should rely on modeling to provide estimates for the entire cohort, supported by subject-derived questionnaire data. c) Assessment of some exposures of interest requires individual measurements of exposures using snapshots of personal and microenvironmental exposures over short periods and/or in selected microenvironments. d) Understanding issues of spatial-temporal correlations of air pollutants, the surrogacy of specific pollutants for components of the complex mixture, and the exposure misclassification inherent in exposure estimates is critical in analysis and interpretation. e) "Usual" temporal, spatial, and physical patterns of activity can be used as modifiers of the exposure/outcome relationships. f) Biomarkers of exposure are useful for evaluation of specific exposures that have multiple routes of exposure. If these lessons are applied, the National Children's Study offers a unique opportunity to assess the adverse effects of air pollution on interrelated health outcomes during the critical early life period.


Perfusion scanning using 99mTc-HMPAO detects early cerebrovascular changes in the diabetic rat.
99mTc-HMPAO is a well-established isotope useful in the detection of regional cerebral blood flow. Diabetes gives rise to arterial atherosclerotic changes that can lead to significant end organ dysfunction, prominently affecting perfusion to the heart, kidneys, eyes and brain. In the current study, we investigated the role of 99mTc-HMPAO cerebral perfusion scans in detecting early vascular changes in the diabetic brain. Cerebral perfusion studies were performed on both control and streptozotocin-(STZ) induced diabetic male Wistar rats. Rat brain imaging using a gamma camera was performed for each group 0.5, 2, 4, and 24 hours post 99mTc-HMPAO injection. Data processing for each cerebral perfusion scan was performed by drawing a region of interest (ROI) circumferentially around the brain (B). Background (BKG) due to signal from the soft tissue of each rat was subtracted. Brain 99mTc-HMPAO uptake minus background counts (net brain counts; NBC) were then compared between the two groups. The NBC (mean +/- SD) for the STZ group were statistically significantly higher (p = 0.0004) than those of the control group at each of the time points studied. 99mTc-HMPAO brain scan may be useful in the detection of early atherosclerotic changes in the diabetic rat brain.


Myocardial viability assessed with fluorodeoxyglucose and PET in patients with Q wave myocardial infarction receiving thrombolysis: relationship to coronary anatomy and ventricular function.
In previously thrombolysed patients, we analysed residual myocardial viability using the PET-FDG technique and correlated its presence and extent to the angiographic appearance of the infarct-related vessel and left ventricular function. Thirty-six patients who had undergone intravenous thromboloysis for acute myocardial infarction 4.8 +/- 7.2 months previously were studied. Coronary angiography, left ventriculography, and assessment of myocardial perfusion and metabolism were all performed within 1 week. All patients exhibited perfusion defects consistent with the clinically identified myocardial infarction site. Residual viability, as assessed by the PET-FDG technique, was present in 53% of cases. The infarct-related coronary artery was patent in 19 (53%) patients (TIMI grade 3, 79%); of the remaining 17 with occluded infarct-related arteries, 11 had collaterals to the infarct area. Significant FDG uptake was observed in 63% of patients with a patent infarct-related artery and in 41% of those with an occluded infarct-related artery. The same study protocol was adopted in a control group of 30 patients with myocardial infarction who did not receive thrombolysis. The number of infarct-related patent vessels was significantly lower in these patients (30 vs 53%) (TIMI grade 3, 56%), but the overall percentage of PET viability was again 53%. Qualitative analysis of the regional perfusion pattern showed that the magnitude and severity of the perfusion defect was similar in the two groups, regardless of the presence or absence of FDG uptake. Global left ventricular function was also similar in the two groups. However, regional wall motion was significantly better in the thrombolysed patients with a patent infarct-related artery than in those who had not received thrombolysis and whose culprit vessel was also patent. In conclusion, the results of our study support the notion that early recanalization of the infarct-related artery is critical for preserving left ventricular function. Although the number of patent infarct-related coronary arteries is greater and left ventricular function is better in successfully thrombolysed patients, the regional metabolic pattern does not apparently correlate with the patency of the infarct-related artery. This suggests that, in "chronic' myocardial infarction, residual tissue viability as assessed by fluorodeoxyglucose uptake does not necessarily correlate with coronary recanalization.


Tyrosine phosphorylation of Kv1.5 is upregulated in intrauterine growth retardation rats with exaggerated pulmonary hypertension.
Intrauterine growth retardation (IUGR) is associated with the development of adult-onset diseases, including pulmonary hypertension. However, the underlying mechanism of the early nutritional insult that results in pulmonary vascular dysfunction later in life is not fully understood. Here, we investigated the role of tyrosine phosphorylation of voltage-gated potassium channel 1.5 (Kv1.5) in this prenatal event that results in exaggerated adult vascular dysfunction. A rat model of chronic hypoxia (2 weeks of hypoxia at 12 weeks old) following IUGR was used to investigate the physiological and structural effect of intrauterine malnutrition on the pulmonary artery by evaluating pulmonary artery systolic pressure and vascular diameter in male rats. Kv1.5 expression and tyrosine phosphorylation in pulmonary artery smooth muscle cells (PASMCs) were determined. We found that IUGR increased mean pulmonary artery pressure and resulted in thicker pulmonary artery smooth muscle layer in 14-week-old rats after 2 weeks of hypoxia, while no difference was observed in normoxia groups. In the PASMCs of IUGR-hypoxia rats, Kv1.5 mRNA and protein expression decreased while that of tyrosine-phosphorylated Kv1.5 significantly increased. These results demonstrate that IUGR leads to exaggerated chronic hypoxia pulmonary arterial hypertension (CH-PAH) in association with decreased Kv1.5 expression in PASMCs. This phenomenon may be mediated by increased tyrosine phosphorylation of Kv1.5 in PASMCs and it provides new insight into the prevention and treatment of IUGR-related CH-PAH.


A new pulsed electric field microreactor: comparison between the laboratory and microtechnology scale.
This paper presents a new microreactor dedicated for pulsed electric field treatment (PEF), which is a pasteurization method that inactivates microorganisms with short electric pulses. The PEF microreactor consists of a flow-through channel with a constriction where the electric field is focussed. Compared to a laboratory-scale setup 25 times lower voltages were needed to obtain the same electric field strength due to the close electrode spacing. A finite element model showed that the electric field intensity is very homogeneous throughout the channel, which is crucial for the pasteurization processes. Experiments where artificial vesicles, loaded with carboxyfluorescein, were electroporated showed that the maximum transmembrane potential adequately described the processes both in the microreactor and the laboratory-scale setup, although the length scales are different. Electroporation started at a transmembrane potential of 0.5 V, reaching a maximum fraction of electroporated vesicles of 51% at a transmembrane potential of 1.5 V. The partial electroporation is not a result of the heterogenity of the vesicles or the electric field. With this new PEF microreactor it is possible to study the PEF process in more detail.


EUREC4A: A Field Campaign to Elucidate the Couplings Between Clouds, Convection and Circulation.
Trade-wind cumuli constitute the cloud type with the highest frequency of occurrence on Earth, and it has been shown that their sensitivity to changing environmental conditions will critically influence the magnitude and pace of future global warming. Research over the last decade has pointed out the importance of the interplay between clouds, convection and circulation in controling this sensitivity. Numerical models represent this interplay in diverse ways, which translates into different responses of trade-cumuli to climate perturbations. Climate models predict that the area covered by shallow cumuli at cloud base is very sensitive to changes in environmental conditions, while process models suggest the opposite. To understand and resolve this contradiction, we propose to organize a field campaign aimed at quantifying the physical properties of trade-cumuli (e.g., cloud fraction and water content) as a function of the large-scale environment. Beyond a better understanding of clouds-circulation coupling processes, the campaign will provide a reference data set that may be used as a benchmark for advancing the modelling and the satellite remote sensing of clouds and circulation. It will also be an opportunity for complementary investigations such as evaluating model convective parameterizations or studying the role of ocean mesoscale eddies in air-sea interactions and convective organization.


Peer-to-peer computing in health-promoting voluntary organizations: a system design analysis.
A large part of the health promotion in today's society is performed as peer-to-peer empowerment in voluntary organisations such as sports clubs, charities, and trade unions. In order to prevent work-related illness and long-term sickness absence, the aim of this study is to explore computer network services for empowerment of employees by peer-to-peer communication. The 'technique trade-off method was used for the analysis of the system design. A Critical Incident Technique questionnaire was distributed to a representative sample of trade union shop stewards (n = 386), and focus-group seminars were arranged where a preliminary set of requirements was discussed. Seven basic requirements were identified and matched to a set of 12 design issues for computer network services, allocating a subset of design issues to each requirement. The conclusion is that the systems design displays an inexpensive and potentially feasible method for peer-to-peer computing in voluntary health-promoting organisations.


Is robotic surgery superior to endoscopic and open surgeries in thyroid cancer?
Endoscopic thyroidectomies have been performed using various approaches, and indications have expanded with the development of new surgical techniques and instruments. Endoscopic thyroid surgery using bilateral axillo-breast approaches have excellent cosmetic results and a symmetrical, optimal operative view. However, because of the two-dimensional view and the nonflexible instruments, these approaches are not easy to use in performing a central lymph node dissection (CND). Robotic surgery has drawn attention as a potentially safe and effective method for treating thyroid cancer. The aim of the present study was to determine whether robotic surgery is superior to endoscopic and open surgery through comparing technical aspects and surgical outcomes. From October 2008 to December 2009, 302 patients had total thyroidectomies and CND with cancer less than 1 cm. Patients were divided into three groups according to operation methods (open group; n = 138), (endo group; n = 95), (robot group; n = 69). Young patients preferred the robotic and endoscopic surgery. The number of retrieved lymph nodes in the open group (4.8 ± 2.8) was not different from the robot group (4.7 ± 2.7) and the endo group (4.6 ± 3.7). The operative time of the robot group was longer than the open and the endo group. The total drain amount in the robot group was more than the open and endo groups; however, there was no difference in the length of hospitalization and complication rates. There were no differences between the open (0.8 ± 2.0) and robot groups (0.8 ± 1.4), but the endo group (2.4 ± 6.3) showed higher postoperative serum thyroglobulin off thyroid hormone (Off-Tg) when compared to the open and robot groups. Robotic surgery was equal to open surgery except with respect to operative time and was superior to endoscopic surgery in Off-Tg levels presenting completeness of the operation in thyroid cancer surgery. Because it has excellent cosmetic results and various technical advantages, it should be considered in young, low-risk patients with thyroid carcinoma less than 1 cm.


HRas signal transduction promotes hepatitis C virus cell entry by triggering assembly of the host tetraspanin receptor complex.
Hepatitis C virus (HCV) entry is dependent on coreceptor complex formation between the tetraspanin superfamily member CD81 and the tight junction protein claudin-1 (CLDN1) on the host cell membrane. The receptor tyrosine kinase EGFR acts as a cofactor for HCV entry by promoting CD81-CLDN1 complex formation via unknown mechanisms. We identify the GTPase HRas, activated downstream of EGFR signaling, as a key host signal transducer for EGFR-mediated HCV entry. Proteomic analysis revealed that HRas associates with tetraspanin CD81, CLDN1, and the previously unrecognized HCV entry cofactors integrin β1 and Ras-related protein Rap2B in hepatocyte membranes. HRas signaling is required for lateral membrane diffusion of CD81, which enables tetraspanin receptor complex assembly. HRas was also found to be relevant for entry of other viruses, including influenza. Our data demonstrate that viruses exploit HRas signaling for cellular entry by compartmentalization of entry factors and receptor trafficking.


The C. elegans rab family: identification, classification and toolkit construction.
Rab monomeric GTPases regulate specific aspects of vesicle transport in eukaryotes including coat recruitment, uncoating, fission, motility, target selection and fusion. Moreover, individual Rab proteins function at specific sites within the cell, for example the ER, golgi and early endosome. Importantly, the localization and function of individual Rab subfamily members are often conserved underscoring the significant contributions that model organisms such as Caenorhabditis elegans can make towards a better understanding of human disease caused by Rab and vesicle trafficking malfunction. With this in mind, a bioinformatics approach was first taken to identify and classify the complete C. elegans Rab family placing individual Rabs into specific subfamilies based on molecular phylogenetics. For genes that were difficult to classify by sequence similarity alone, we did a comparative analysis of intron position among specific subfamilies from yeast to humans. This two-pronged approach allowed the classification of 30 out of 31 C. elegans Rab proteins identified here including Rab31/Rab50, a likely member of the last eukaryotic common ancestor (LECA). Second, a molecular toolset was created to facilitate research on biological processes that involve Rab proteins. Specifically, we used Gateway-compatible C. elegans ORFeome clones as starting material to create 44 full-length, sequence-verified, dominant-negative (DN) and constitutive active (CA) rab open reading frames (ORFs). Development of this toolset provided independent research projects for students enrolled in a research-based molecular techniques course at California State University, East Bay (CSUEB).


A novel recurrent mutation in MITF predisposes to familial and sporadic melanoma.
So far, two genes associated with familial melanoma have been identified, accounting for a minority of genetic risk in families. Mutations in CDKN2A account for approximately 40% of familial cases, and predisposing mutations in CDK4 have been reported in a very small number of melanoma kindreds. Here we report the whole-genome sequencing of probands from several melanoma families, which we performed in order to identify other genes associated with familial melanoma. We identify one individual carrying a novel germline variant (coding DNA sequence c.G1075A; protein sequence p.E318K; rs149617956) in the melanoma-lineage-specific oncogene microphthalmia-associated transcription factor (MITF). Although the variant co-segregated with melanoma in some but not all cases in the family, linkage analysis of 31 families subsequently identified to carry the variant generated a log of odds (lod) score of 2.7 under a dominant model, indicating E318K as a possible intermediate risk variant. Consistent with this, the E318K variant was significantly associated with melanoma in a large Australian case-control sample. Likewise, it was similarly associated in an independent case-control sample from the United Kingdom. In the Australian sample, the variant allele was significantly over-represented in cases with a family history of melanoma, multiple primary melanomas, or both. The variant allele was also associated with increased naevus count and non-blue eye colour. Functional analysis of E318K showed that MITF encoded by the variant allele had impaired sumoylation and differentially regulated several MITF targets. These data indicate that MITF is a melanoma-predisposition gene and highlight the utility of whole-genome sequencing to identify novel rare variants associated with disease susceptibility.


Mechanisms of NAFLD development and therapeutic strategies.
There has been a rise in the prevalence of nonalcoholic fatty liver disease (NAFLD), paralleling a worldwide increase in diabetes and metabolic syndrome. NAFLD, a continuum of liver abnormalities from nonalcoholic fatty liver (NAFL) to nonalcoholic steatohepatitis (NASH), has a variable course but can lead to cirrhosis and liver cancer. Here we review the pathogenic and clinical features of NAFLD, its major comorbidities, clinical progression and risk of complications and in vitro and animal models of NAFLD enabling refinement of therapeutic targets that can accelerate drug development. We also discuss evolving principles of clinical trial design to evaluate drug efficacy and the emerging targets for drug development that involve either single agents or combination therapies intended to arrest or reverse disease progression.


Quantitative MALDI-MS/MS assay for serum cortisol through charged derivatization.
Cortisol (CRT), the main glucocorticoid in humans, plays a crucial role in many physiological processes, therefore, the measurement of its serum level is of great clinical significance. Matrix-assisted laser desorption/ionization-tandem mass spectrometry (MALDI-MS/MS) might be an effective approach for the quantification of CRT in serum due to its attractive properties, i.e., high specificity, ease of use, ruggedness and rapid analysis. In this study, we developed a method to quantify the serum CRT by MALDI-MS/MS. This method employed the derivatization using a Girard-type reagent, 1-(hydrazinocarbonylmethyl)isoquinolinium chloride, which compensated for the lack of sensitivity. This method enabled the reproducible quantification of the serum CRT using a 20-μL sample (intra- and inter-assay relative standard deviations, 7.4% or lower), and the measurable range was 25-500 ng/mL. The serum CRT concentrations determined by the newly-developed MALDI-MS/MS method agreed well with those by liquid chromatography/electrospray ionization-MS/MS or electrochemiluminescence immunoassay. The MALDI-MS/MS method was used for the analysis of peripheral venous serum samples of healthy subjects and adrenal venous serum samples of patients with primary aldosteronism, and satisfactory results were obtained.


Resilience in Vulnerable Populations With Type 2 Diabetes Mellitus and Hypertension: A Systematic Review and Meta-analysis.
Patients with chronic conditions and limited access to health care experience stressful challenges resulting from the burden of managing both their conditions and their daily life demands. Resilience provides a mechanism of adapting to stressful experiences. We conducted a systematic review and meta-analysis to synthesize the evidence about interventions to enhance resiliency in managing hypertension or type 2 diabetes in vulnerable populations and to assess the efficacy of these interventions on clinical outcomes. We searched multiple databases from early inception through February 2015 including randomized controlled trials that enrolled patients with type 2 diabetes or hypertension. All interventions that targeted resilience in vulnerable populations were included. Data were synthesized to describe the characteristics and efficacy of resiliency interventions. We pooled the total effects by calculating standardized mean difference using the random-effects model. The final search yielded 17 studies. All studies were conducted in the United States and generally targeted minority participants. Resiliency interventions used diverse strategies; discussion groups or workshops were the most common approach. Interventions aimed at enhancing the resiliency of patients from vulnerable groups are diverse. Outcomes were not fully conclusive. There was some evidence that resiliency interventions had a positive effect on hemoglobin A1C levels but not blood pressure. The incorporation of resiliency-oriented interventions into the arsenal of preventing and managing chronic conditions appears to be an opportunity that remains to be better investigated and exploited, and there is need to pursue further understanding of the core components of any intervention that claims to enhance resilience.


Environmental effects and aquatic organisms: investigations of molecular mechanisms of carcinogenesis.
Cancers of the reproductive system are among the leading causes of mortality in women in the United States. While both genetic and environmental factors have been implicated in their etiology, the extent of the contribution of environmental factors to human diseases remains controversial. To better address the role of environmental exposures in cancer etiology, there has been an increasing focus on the development of nontraditional, environmentally relevant models. Our research involves the development of one such model. Gonadal tumors have been described in the softshell clam (Mya arenaria) in Maine and the hardshell clam (Mercenaria spp.) from Florida. Prevalence of these tumors is as high as 40% in some populations in eastern Maine and 60% in some areas along the Indian River in Florida. The average tumor prevalence in Maine and Florida is approximately 20 and 11%, respectively. An association has been suggested between the use of herbicides and the incidence of gonadal tumors in the softshell clam in Maine. The role of environmental exposures in the development of the tumors in Mercenaria in Florida is unknown; however, there is evidence that genetic factors may contribute to its etiology. Epidemiologic studies of human populations in these same areas show a higher than average mortality rate due to cancers of the reproductive system in women, including both ovarian and breast cancer. The relationship, if any, among these observations is unknown. Our studies on the molecular basis of this disease in clams may provide additional information on environmental exposures and their possible link to cancer in clams and other organisms, including humans.


Coping and acceptance: the greatest challenge for veterans with intestinal stomas.
Intestinal stomas (ostomies) create challenges for veterans. The goal of this qualitative analysis was to understand better patients' perspectives regarding their greatest challenge. Ostomates at three Veterans Affairs locations were surveyed using the modified City of Hope Quality of Life-Ostomy questionnaire that contained an open-ended request for respondents to describe their greatest challenge. The response rate was 51% (239 of 467); 68% (163 of 239) completed the open-ended item. Content analysis was performed by an experienced qualitative research team. Coping and acceptance were the most commonly addressed themes. The most frequently expressed issues and advice were related to a need for positive thinking and insight regarding adjustment over time. Coping strategies included the use of humor, recognition of positive changes resulting from the stoma, and normalization of life with an ostomy. Coping and acceptance are common themes described by veterans with an intestinal stoma. Health-care providers can assist veterans by utilizing ostomate self-management strategies, experience, and advice.


Audiovisual semantic interactions between linguistic and nonlinguistic stimuli: The time-courses and categorical specificity.
We examined the time-courses and categorical specificity of the crossmodal semantic congruency effects elicited by naturalistic sounds and spoken words on the processing of visual pictures (Experiment 1) and printed words (Experiment 2). Auditory cues were presented at 7 different stimulus onset asynchronies (SOAs) with respect to the visual targets, and participants made speeded categorization judgments (living vs. nonliving). Three common effects were observed across 2 experiments: Both naturalistic sounds and spoken words induced a slowly emerging congruency effect when leading by 250 ms or more in the congruent compared with the incongruent condition, and a rapidly emerging inhibitory effect when leading by 250 ms or less in the incongruent condition as opposed to the noise condition. Only spoken words that did not match the visual targets elicited an additional inhibitory effect when leading by 100 ms or when presented simultaneously. Compared with nonlinguistic stimuli, the crossmodal congruency effects associated with linguistic stimuli occurred over a wider range of SOAs and occurred at a more specific level of the category hierarchy (i.e., the basic level) than was required by the task. A comprehensive framework is proposed to provide a dynamic view regarding how meaning is extracted during the processing of visual or auditory linguistic and nonlinguistic stimuli, therefore contributing to our understanding of multisensory semantic processing in humans. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Pharmacist-provided immunization compensation and recognition: white paper summarizing APhA/AMCP stakeholder meeting.
To identify the current challenges and opportunities in compensation and recognition for pharmacist-provided immunizations across the lifespan and to establish guiding principles for pharmacist-provided immunization compensation and recognition. 22 stakeholders gathered on June 29, 2011, at the American Pharmacists Association (APhA) headquarters in Washington, DC, for a meeting on immunization compensation that was convened by APhA and the Academy of Managed Care Pharmacy. Participants included representatives from community pharmacy practices (chain, grocery, and independent), employers, national consumer health and advocacy organizations, national pharmacy and public health organizations, health plan representatives, pharmacy benefit managers, and health information technology, standards, and safety organizations. Key immunization leaders from TRICARE Management Activity, the Centers for Medicare & Medicaid Services, the National Vaccine Program Office of the Department of Health & Human Services, and the Centers for Disease Control and Prevention (CDC) also participated in the meeting. The increased numbers of pharmacists providing vaccination services and the availability of pharmacist-provided immunizations to populations in need of vaccines has continued to increase. This has resulted in a rise in the percentage of patients who receive vaccines at pharmacies. Pharmacists are now working to lever-age their ability to identify people with key risk factors (e.g., diabetes, heart disease or previous myocardial infarction), encourage them to receive their CDC-recommended vaccinations, and administer the required vaccine. Challenges and opportunities in compensation and recognition for pharmacist-provided immunizations across the adult lifespan persist. Variability in state practice acts, reimbursement and compensation processes and systems, and mechanisms for documentation of vaccine services create substantial differences in how pharmacist-provided immunizations are delivered throughout the United States. Pharmacist-provided immunizations are clinically sound, are cost effective, are readily accessible, and support our nation's public health goals. Pharmacists have demonstrated that patient vaccination rates have improved through expansion of pharmacist-provided immunizations. The profession should continue efforts to collaborate with other immunization stakeholders and expand a pharmacist scope of practice that is built around a uniform and recognized standard of immunization provision and that supports the provision of all CDC-recommended vaccines through pharmacy-provided immunizations.


[Effects of autologous mesenchymal stem cells transfected with heme oxygenase-1 gene transplantation on ischemic Swine hearts].
To observe the effect of intracoronary transfer of autologous HO-1 overexpressed MSCs in porcine model of myocardial ischemia (1 h)/reperfusion. Apoptosis was assayed and cytokine concentrations in supernatant were measured in cells exposed to hypoxia-reoxygen in vitro. In vivo, Chinese male mini-pigs were allocated to the following treatment groups: control group (saline), MSCs group (MSCs), MSCs transfected with pcDNA3.1-nHO-1 (HO-1-MSCs). 1 x 10(7) of autologous stem cells or identical volume of saline was injected intracoronary into porcine hearts 1 h after ischemia. MRI assay and postmortem analysis were assessed 3 months after stem cell transplantation. In vitro, cell apoptosis rate post hypoxia-reoxygen was significantly reduced in HO-1-MSCs group (30.30% +/- 7.64%) compared with that in MSCs group (56.93% +/- 4.68%, P < 0.001) and LacZ-MSCs group (55.88% +/- 4.38%, P < 0.001), VEGF was also significantly upregulated in HO-1-MSCs group [(768.44 +/- 78.38) pg/ml] compared with that in MSCs group [(555.27 +/- 67.67) pg/ml, P < 0.001] and LacZ-MSCs group [(522.97 +/- 71.45) pg/ml, P < 0.001]. In vivo, cardiac function was significantly improved in both MSCs transplantation groups compared to saline group (all P < 0.05 vs.saline) and the left ventricular ejection fraction was significantly higher in HO-1-MSCs group compared with that in MSCs group at 3 months after transplantation (53.50% +/- 2.09% vs. 49.54% +/- 2.74%, P = 0.017), capillary density in the peri-infarct area was also significantly higher in HO-1-MSC group than that in MSCs group [(14.59 +/- 2.39)/HPF vs. (11.78 +/- 2.48)/HPF, P = 0.033]. Efficacy of HO-1 overexpressed MSCs on improving cardiac function and promoting angiogenesis was greater than those by MSCs in this porcine ischemia/reperfusion model.


Vertebral fracture: epidemiology, impact and use of DXA vertebral fracture assessment in fracture liaison services.
Vertebral fractures are independent risk factors for vertebral and nonvertebral fractures. Since vertebral fractures are often missed, the relatively new introduction of vertebral fracture assessment (VFA) for imaging of the lateral spine during DXA-measurement of the spine and hips may contribute to detect vertebral fractures. We advocate performing a VFA in all patients with a recent fracture visiting a fracture liaison service (FLS). Fracture liaison services (FLS) are important service models for delivering secondary fracture prevention for older adults presenting with a fragility fracture. While commonly age, clinical risk factors (including fracture site and number of prior fracture) and BMD play a crucial role in determining fracture risk and indications for treatment with antiosteoporosis medications, prevalent vertebral fractures usually remain undetected. However, vertebral fractures are important independent risk factors for future vertebral and nonvertebral fractures. A development of the DXA technology, vertebral fracture assessment (VFA), allows for assessment of the lateral spine during the regular DXA bone mineral density measurement of the lumbar spine and hips. Recent approaches to the stratification of antiosteoporosis medication type according to baseline fracture risk, and differences by age in the indication for treatment by prior fracture mean that additional information from VFA may influence initiation and type of treatment. Furthermore, knowledge of baseline vertebral fractures allows reliable definition of incident vertebral fracture events during treatment, which may modify the approach to therapy. In this manuscript, we will discuss the epidemiology and clinical significance of vertebral fractures, the different methods of detecting vertebral fractures, and the rationale for, and implications of, use of VFA routinely in FLS. • Vertebral fracture assessment is a tool available on modern DXA instruments and has proven ability to detect vertebral fractures, the majority of which occur without a fall and without the signs and symptoms of an acute fracture. • Most osteoporosis guidelines internationally suggest that treatment with antiosteoporosis medications should be considered for older individuals (e.g., 65 years +) with a recent low trauma fracture without the need for DXA. • Younger individuals postfracture may be risk-assessed on the basis of FRAX® probability including DXA and associated treatment thresholds. • Future fracture risk is markedly influenced by both site, number, severity, and recency of prior fracture; awareness of baseline vertebral fractures facilitates definition of true incident vertebral fracture events occurring during antiosteoporosis treatment. • Detection of previously clinically silent vertebral fractures, defining site of prior fracture, might alter treatment decisions in younger or older FLS patients, consistent with recent IOF-ESCEO guidance on baseline-risk-stratified therapy, and provides a reliable baseline from which to define new, potentially therapy-altering, vertebral fracture events.


Unique human immune signature of Ebola virus disease in Guinea.
Despite the magnitude of the Ebola virus disease (EVD) outbreak in West Africa, there is still a fundamental lack of knowledge about the pathophysiology of EVD. In particular, very little is known about human immune responses to Ebola virus. Here we evaluate the physiology of the human T cell immune response in EVD patients at the time of admission to the Ebola Treatment Center in Guinea, and longitudinally until discharge or death. Through the use of multiparametric flow cytometry established by the European Mobile Laboratory in the field, we identify an immune signature that is unique in EVD fatalities. Fatal EVD was characterized by a high percentage of CD4(+) and CD8(+) T cells expressing the inhibitory molecules CTLA-4 and PD-1, which correlated with elevated inflammatory markers and high virus load. Conversely, surviving individuals showed significantly lower expression of CTLA-4 and PD-1 as well as lower inflammation, despite comparable overall T cell activation. Concomitant with virus clearance, survivors mounted a robust Ebola-virus-specific T cell response. Our findings suggest that dysregulation of the T cell response is a key component of EVD pathophysiology.


The diploid genome sequence of an individual human.
Presented here is a genome sequence of an individual human. It was produced from approximately 32 million random DNA fragments, sequenced by Sanger dideoxy technology and assembled into 4,528 scaffolds, comprising 2,810 million bases (Mb) of contiguous sequence with approximately 7.5-fold coverage for any given region. We developed a modified version of the Celera assembler to facilitate the identification and comparison of alternate alleles within this individual diploid genome. Comparison of this genome and the National Center for Biotechnology Information human reference assembly revealed more than 4.1 million DNA variants, encompassing 12.3 Mb. These variants (of which 1,288,319 were novel) included 3,213,401 single nucleotide polymorphisms (SNPs), 53,823 block substitutions (2-206 bp), 292,102 heterozygous insertion/deletion events (indels)(1-571 bp), 559,473 homozygous indels (1-82,711 bp), 90 inversions, as well as numerous segmental duplications and copy number variation regions. Non-SNP DNA variation accounts for 22% of all events identified in the donor, however they involve 74% of all variant bases. This suggests an important role for non-SNP genetic alterations in defining the diploid genome structure. Moreover, 44% of genes were heterozygous for one or more variants. Using a novel haplotype assembly strategy, we were able to span 1.5 Gb of genome sequence in segments >200 kb, providing further precision to the diploid nature of the genome. These data depict a definitive molecular portrait of a diploid human genome that provides a starting point for future genome comparisons and enables an era of individualized genomic information.


Knockin' on pollen's door: live cell imaging of early polarization events in germinating Arabidopsis pollen.
Pollen tubes are an excellent system for studying the cellular dynamics and complex signaling pathways that coordinate polarized tip growth. Although several signaling mechanisms acting in the tip-growing pollen tube have been described, our knowledge on the subcellular and molecular events during pollen germination and growth site selection at the pollen plasma membrane is rather scarce. To simultaneously track germinating pollen from up to 12 genetically different plants we developed an inexpensive and easy mounting technique, suitable for every standard microscope setup. We performed high magnification live-cell imaging during Arabidopsis pollen activation, germination, and the establishment of pollen tube tip growth by using fluorescent marker lines labeling either the pollen cytoplasm, vesicles, the actin cytoskeleton or the sperm cell nuclei and membranes. Our studies revealed distinctive vesicle and F-actin polarization during pollen activation and characteristic growth kinetics during pollen germination and pollen tube formation. Initially, the germinating Arabidopsis pollen tube grows slowly and forms a uniform roundish bulge, followed by a transition phase with vesicles heavily accumulating at the growth site before switching to rapid tip growth. Furthermore, we found the two sperm cells to be transported into the pollen tube after the phase of rapid tip growth has been initiated. The method presented here is suitable to quantitatively study subcellular events during Arabidopsis pollen germination and growth, and for the detailed analysis of pollen mutants with respect to pollen polarization, bulging, or growth site selection at the pollen plasma membrane.


A micro-level event-centered approach to investigating armed conflict and population responses.
In this article, we construct and test a micro-level event-centered approach to the study of armed conflict and behavioral responses in the general population. Event-centered approaches have been successfully used in the macro-political study of armed conflict but have not yet been adopted in micro-behavioral studies. The micro-level event-centered approach that we advocate here includes decomposition of a conflict into discrete political and violent events, examination of the mechanisms through which they affect behavior, and consideration of differential risks within the population. We focus on two mechanisms: instability and threat of harm. We test this approach empirically in the context of the recent decade-long armed conflict in Nepal, using detailed measurements of conflict-related events and a longitudinal study of first migration, first marriage, and first contraceptive use. Results demonstrate that different conflict-related events independently shaped migration, marriage, and childbearing and that they can simultaneously influence behaviors in opposing directions. We find that violent events increased migration, but political events slowed migration. Both violent and political events increased marriage and contraceptive use net of migration. Overall, this micro-level event-centered approach yields a significant advance for the study of how armed conflict affects civilian behavioral responses.


Comparison of Postoperative Outcomes of Laparoscopic vs Open Inguinal Hernia Repair.
Advocates of laparoscopic surgery argue that all inguinal hernias, including initial and unilateral ones, should be repaired laparoscopically. Prior work suggests outcomes of open repair are improved by using local rather than general anesthesia, but no prior studies have compared laparoscopic surgery with open repair under local anesthesia. To evaluate postoperative outcomes of open inguinal hernia repair under general or local anesthesia compared with laparoscopic repair. This retrospective cohort study identified 107 073 patients in the Veterans Affairs Surgical Quality Improvement Program database who underwent unilateral initial inguinal hernia repair from 1998 to 2019. Data were analyzed from October 2021 to March 2022. Patients were divided into 3 groups for comparison: (1) open repair with local anesthesia (n = 22 333), (2) open repair with general anesthesia (n = 75 104), and (3) laparoscopic repair with general anesthesia (n = 9636). Operative time and postoperative morbidity were compared using quantile regression and inverse probability propensity weighting. A 2-stage least-squares regression and probabilistic sensitivity analysis was used to quantify and address bias from unmeasured confounding in this observational study. Of 107 073 included patients, 106 529 (99.5%) were men, and the median (IQR) age was 63 (55-71) years. Compared with open repair with general anesthesia, laparoscopic repair was associated with a nonsignificant 0.15% (95% CI, -0.39 to 0.09; P = .22) reduction in postoperative complications. There was no significant difference in complications between laparoscopic surgery and open repair with local anesthesia (-0.05%; 95% CI, -0.34 to 0.28; P = .70). Operative time was similar for the laparoscopic and open general anesthesia groups (4.31 minutes; 95% CI, 0.45-8.57; P = .048), but operative times were significantly longer for laparoscopic compared with open repair under local anesthesia (10.42 minutes; 95% CI, 5.80-15.05; P < .001). Sensitivity analysis and 2-stage least-squares regression demonstrated that these findings were robust to bias from unmeasured confounding. In this study, laparoscopic and open repair with local anesthesia were reasonable options for patients with initial unilateral inguinal hernias, and the decision should be made considering both patient and surgeon factors.


Excretion of host immunoglobulin in tick saliva and detection of IgG-binding proteins in tick haemolymph and salivary glands.
Host immunoglobulin G (IgG) crossed the gut wall into the haemocoel of adult Rhipicephalus appendiculatus female ticks when they fed on guinea-pigs. Guinea-pig IgG was also found in saliva of the feeding ticks. The concentration and antibody activity of IgG in haemolymph, salivary gland extract (SGE) and saliva at different stages of tick feeding were detected by enzyme-linked immunoassay. Specific activity of the IgG in tick samples was determined by feeding ticks on guinea-pigs which were immunized with killed Escherichia coli: 35-42% of the antibody activity in guinea-pig immune serum remained in the tick samples. The high relative concentration of IgG in tick saliva at later stages of feeding suggests that the tick may have a mechanism for getting rid of foreign proteins via the salivary gland. Such a mechanism could involve IgG binding proteins (IGBPs) which were found in both haemolymph and SGE of female ticks at day 6 of feeding using a guinea-pig IgG-agarose affinity column. In female ticks, the M(r) of IGBPs in SGE (23 and 57 kDa) were less than those in haemolymph (78 and > 100 kDa). The existence of IGBPs in both the tick salivary gland and haemolymph indicate that haemolymph and salivary gland cooperate to remove foreign proteins, e.g. host immunoglobulin, from the body during feeding. This mechanism may be a part of the tick self-defence system.


Predictive factors of neoplastic gallbladder polyps: Outcomes of 278 patients.
Distinguishing between neoplastic and nonneoplastic gallbladder polyps (GBPs) in the preoperative workup is of great importance for appropriate treatment. The present study aimed to investigate the characteristics of GBPs and to determine potential predictive factors of neoplastic polyps. The data of 278 patients who were confirmed to have GBPs through laparoscopic cholecystectomy were retrospectively analyzed. Polyps were classified as nonneoplastic and neoplastic GBPs, according to histopathological diagnoses. All clinicopathological characteristics were compared between these two groups. There were 264 (95%) nonneoplastic GBPs and 14 (5%) neoplastic GBPs. In univariate analysis, there were significant differences in age with a cutoff value of 60 years (p=0.002), polyp size (p<0.001), number of polyps (p=0.014), and polyp morphology (p<0.001) between the groups. Multivariate analysis showed that solitary polyp (p<0.001) and sessile morphology (p<0.001) were the independent predictors of neoplastic GBPs. Receiver-operating characteristic curve analysis of three cut-off values of polyp sizes (6, 10, and 15 mm) indicated that a polyp size of 10 mm had the highest area under curve (0.942). Age above 60 years, solitary polyps larger than 1 cm, and sessile morphology are associated with an increased risk of neoplasia in GBP. Therefore, these characteristics should be considered in the management of GBPs to reduce the incidence of unnecessary surgeries and to prevent delays in the treatment of a possible cancer.


Decreased interactions of mutant muscle LIM protein (MLP) with N-RAP and alpha-actinin and their implication for hypertrophic cardiomyopathy.
Previous work has shown that mutations in muscle LIM protein (MLP) can cause hypertrophic cardiomyopathy (HCM). In order to gain an insight into the molecular basis of the disease phenotype, we analysed the binding characteristics of wild-type MLP and of the (C58G) mutant MLP that causes hypertrophic cardiomyopathy. We show that MLP can form a ternary complex with two of its previously documented myofibrillar ligand proteins, N-RAP and alpha-actinin, which indicates the presence of distinct, non-overlapping binding sites. Our data also show that, in comparison to wild-type MLP, the capacity of the mutated MLP protein to bind both N-RAP and alpha-actinin is significantly decreased. In addition, this single point mutation prevents zinc coordination and proper folding of the second zinc-finger in the first LIM domain, which consequently renders the protein less stable and more susceptible to proteolysis. The molecular basis for HCM-causing mutations in the MLP gene might therefore be an alteration in the equilibrium of interactions of the ternary complex MLP-N-RAP-alpha-actinin. This assumption is supported by the previous observation that in the pathological situation accompanied by MLP down regulation, cardiomyocytes try to compensate for the decreased stability of MLP protein by increasing the expression of its ligand N-RAP, which might finally result in the development of myocyte disarray that is characteristic of this disease.


E-Cigarettes: A Review of New Trends in Cannabis Use.
The emergence of electronic cigarettes (e-cigs) has given cannabis smokers a new method of inhaling cannabinoids. E-cigs differ from traditional marijuana cigarettes in several respects. First, it is assumed that vaporizing cannabinoids at lower temperatures is safer because it produces smaller amounts of toxic substances than the hot combustion of a marijuana cigarette. Recreational cannabis users can discretely "vape" deodorized cannabis extracts with minimal annoyance to the people around them and less chance of detection. There are nevertheless several drawbacks worth mentioning: although manufacturing commercial (or homemade) cannabinoid-enriched electronic liquids (e-liquids) requires lengthy, complex processing, some are readily on the Internet despite their lack of quality control, expiry date, and conditions of preservation and, above all, any toxicological and clinical assessment. Besides these safety problems, the regulatory situation surrounding e-liquids is often unclear. More simply ground cannabis flowering heads or concentrated, oily THC extracts (such as butane honey oil or BHO) can be vaped in specially designed, pen-sized marijuana vaporizers. Analysis of a commercial e-liquid rich in cannabidiol showed that it contained a smaller dose of active ingredient than advertised; testing our laboratory-made, purified BHO, however, confirmed that it could be vaped in an e-cig to deliver a psychoactive dose of THC. The health consequences specific to vaping these cannabis preparations remain largely unknown and speculative due to the absence of comprehensive, robust scientific studies. The most significant health concerns involve the vaping of cannabinoids by children and teenagers. E-cigs could provide an alternative gateway to cannabis use for young people. Furthermore, vaping cannabinoids could lead to environmental and passive contamination.


Chromium supplementation in overweight and obesity: a systematic review and meta-analysis of randomized clinical trials.
The increased prevalence of obesity has made the use of dietary supplements as weight reducing agents highly popular, but their efficacy has not been proven. One such supplement is chromium. The purpose of this review was to evaluate the evidence for or against the efficacy of chromium supplementation in overweight and obese individuals. Electronic searches were conducted in Medline, Embase, Amed and The Cochrane Library. The bibliographies of located articles were also searched. No age, gender or language restrictions were imposed. The reporting quality of identified randomized clinical trials (RCTs) was assessed using a methodological checklist adapted from the Consolidated Standard of Reporting Trials Statement and Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Thirty-nine trials were identified and 20 were included. There were variations in reporting quality of included studies. A meta-analysis of 11 studies showed a statistically significant difference in weight loss favouring chromium over placebo (mean difference (MD): -0.50 kg; 95% confidence interval (CI): -0.97, -0.03). There was a high statistical heterogeneity. Adverse events included watery stools, vertigo, headaches and urticaria. The evidence from available RCTs shows that chromium supplementation generates statistically significant reductions in body weight. The magnitude of the effect is small, and the clinical relevance is uncertain. Future trials should last at least 16 weeks and greater uniformity in the measuring and assessment tools for body composition is recommended.