Expression of miR-34 is lost in colon cancer which can be re-expressed by a novel agent CDF.
Colorectal Cancer (CRC) is one of the leading causes of death worldwide. Numerous cellular events, including deregulated expression of microRNAs (miRNAs), specifically the family of miR-34 consisting of miR-34a, b and c, is known to regulate the processes of growth and metastasis. We evaluated the expression of miR-34 in formalin-fixed paraffin-embedded (FFPE) human colon cancer tissue specimens compared to normal colonic mucosa. Moreover, we also assessed the expression of miR-34 in colon cancer cell lines treated with our newly developed synthetic analogue of curcumin referred as difluorinated curcumin (CDF) compared to well known inhibitor of methyl transferase. We found that the expression of miR-34a and miR-34c was down-regulated in colon cancer specimens compared to normal colonic mucosa and the loss of expression was also consistent with data from colon cancer cell lines. This down-regulation was attributed to promoter hypermethylation, because we found that the treatment of colon cancer cells with 5-aza-2´-deoxycytidine, a methyltransferase inhibitor, markedly induced the levels of miR-34a and miR-34c expression. Likewise, CDF was very effective in the re-expression of miR-34a and miR-34c, which was consistent with inhibition of cell growth of both chemo-sensitive and chemo-resistant colon cancer cells. The re-expression of miR-34 led to a marked reduction in the expression of its target gene, Notch-1. The loss of expression of miR-34 in colon cancer is in part due to promoter hypermethylation of miR-34, which can be re-expressed with our novel agent CDF, suggesting that CDF could be a novel demethylating agent for restoring the expression of miR-34 family, and thus CDF could become a newer therapeutic agent for the treatment of colon cancer.


The role of cancer metabolism in defining the success of oncolytic viro-immunotherapy.
Oncolytic viruses infect, replicate in, and kill cancer cells selectively without harming normal cells. The rapidly expanding clinical development of oncolytic virotherapy is an exciting interdisciplinary field that provides insights into virology, oncology, and immunotherapy. Recent years have seen greater focus on rational design of cancer-selective viruses together with strategies to exploit their immunostimulatory capabilities, ultimately to develop powerful oncolytic cancer vaccines. However, despite great interest in the field, many important experiments are still conducted under optimum conditions in vitro, with many nutrients present in excess and with cellular stress kept to a minimum. Whilst this provides a convenient platform for cell culture, it bears little relation to the typical conditions found within a tumour in vivo, where cells are often subject to a range of metabolic and environmental stresses. Viral infection and cancer will both lead to production of metabolites that are also not present in media in vitro. Understanding how oncolytic viruses interact with cells exposed to more representative metabolic conditions in vitro represents an under-explored area of study that could provide valuable insight into the intelligent design of superior oncolytic viruses and help bridge the gap between bench and bedside. This review summarises the major metabolic pathways altered in cancer cells, during viral infection and highlights possible targets for future studies.


Cell-specific accumulation of maize phosphoenolpyruvate carboxylase is correlated with demethylation at a specific site greater than 3 kb upstream of the gene.
Development of the C4 photosynthetic pathway relies upon the cell-specific accumulation of photosynthetic enzymes. Although the molecular basis of this cell-specific gene expression is not known, regulation appears to be exerted at the level of transcript accumulation. We have investigated the relationship between gene expression patterns and DNA methylation for genes of two of the C4 photosynthetic enzymes, ribulose bisphosphate carboxylase (RuBPCase) and phosphoenolpyruvate carboxylase (PEPCase). We found no correlation between methylation state and gene expression for either the large subunit or a small subunit gene of RuBPCase. In contrast, demethylation of a specific site 5' to the PEPCase gene was correlated with the light-induced, cell-specific accumulation of PEPCase mRNA. This differentially methylated site is positioned at great distance (greater than 3 kb) from the start of transcription. This observation is made more interesting by the fact that the immediate 5' region of the gene, and some of the coding region, represents an unmethylated CpG island. Such islands are normally associated with constitutively expressed genes.


Aesthetic surgery and Google: ubiquitous, unregulated and enticing websites for patients considering cosmetic surgery.
Patient safety is a fundamental issue in aesthetic surgery. In an attempt to improve safety, the Department of Health (DoH) and Professor Sir Bruce Keogh published a review in 2013 of the regulation of cosmetic interventions. Proposals included: (1) Banning free consultations; (2) Restricting time-limited promotional deals; (3) Two-stage written pre-operative consent; (4) Consultations with a medical professional rather than a sales 'consultant'. The Cosmetic Surgical Practice Working Party (CSWP) recommended a two week "cooling off" period before surgery. This study quantified compliance with the above national initiatives by aesthetic surgery providers in the UK. To replicate a patient searching for aesthetic surgery providers, "cosmetic surgery UK" was searched via Google. The top fifty websites of aesthetic surgery providers were included in the study. Websites were analysed for compliance with the DoH Keogh and CSWP recommendations. When clarification was required, aesthetic surgery providers were contacted via telephone. Pearson's Chi-squared test compared actual compliance with national recommendations of full compliance. Fifty cosmetic surgery providers in the UK entered the study. Consultations with the operating surgeon occurred in 90% of cases. Mean compliance with all parameters from the national guidelines was 41%, significantly less than the desired level of full compliance (P < 0.001). The majority offered free consultations (54%) and promotional deals (52%), of which 27% were time limited. No provider stipulated compliance with two stages of signed consent. This study demonstrated low compliance with national guidelines for aesthetic surgery. Aggressive sales techniques and enticing offers by aesthetic surgery providers were widespread. Statutory government guidelines on aesthetic surgery and increased public awareness into potential risks from inappropriate cosmetic surgery may improve patient decision making and safety.


Intraoperative perfusion mapping with laser-assisted indocyanine green imaging can predict and prevent complications in immediate breast reconstruction.
Immediate breast reconstruction results in a superior cosmetic outcome. However, immediate breast reconstruction using both prosthetic and autologous techniques is associated with significantly higher complication rates than delayed procedures. These early postoperative complications are usually related to unrecognized ischemia of mastectomy skin and/or inadequate perfusion of autologous tissue used for reconstruction. Aside from clinical experience, there are no reliable tools to assist the novice surgeon with intraoperative assessment of tissue viability. Laser-assisted indocyanine green imaging was applied to determine and map tissue perfusion. Indocyanine green perfusion mapping was used in 24 consecutive breast reconstructions to define the perfusion of both mastectomy skin and autologous tissue. Areas of inadequate perfusion were then removed at the time of surgery. Postoperative complications occurring within 90 days after surgery were reviewed. In 24 consecutive breast reconstruction (16 tissue expanders, two latissimus dorsi flaps, and six deep inferior epigastric perforator/superficial inferior epigastric arteries), there was a 4 percent complication rate. Intraoperatively, the use of indocyanine green imaging allowed all poorly perfused skin to be removed completely in each case, minimizing the incidence of mastectomy flap necrosis, partial necrosis of autologous tissue, and impaired healing. For autologous reconstruction, patency of anastomoses could also be confirmed. This complication rate was significantly less than the 15.1 percent complication rate observed in 206 reconstructions in the previous consecutive 148 patients (p < 0.01) with similar demographics and risk factors. This early experience demonstrates an increased accuracy in predicting tissue necrosis (mastectomy flap, autologous tissue) as guided by indocyanine green imaging. Further prospective studies are warranted to quantify whether this technology can reduce health care costs by preventing complications in immediate breast reconstruction.


Associations between illness duration and health-related quality of life in specified mental and physical chronic health conditions: results from a population-based survey.
We compared health-related quality of life (HRQOL) in incident (≤1 year since diagnosis), mid-term (>1-5 years since diagnosis), and long-term (>5 years since diagnosis) cases of mental and physical chronic illness with the general population and assessed the modifying effects of age and gender on the association between HRQOL and illness duration. Data from the 2007 Australian National Health and Mental Wellbeing Survey were used. HRQOL was captured by the Assessment of Quality of Life Scale 4D. Multivariable linear regression analyses compared HRQOL of individuals with different duration of illnesses with those who did not have the condition of interest. The 8841 survey respondents were aged 16-85 years (median 43 years, 50.3% female). For the overall sample, worse HRQOL was associated with incident (P = 0.049) and mid-term (P = 0.036) stroke and long-term depression (P < 0.001) and anxiety (P = 0.001). Age had moderating effect on the associations between HRQOL and duration of asthma (P < 0.001), arthritis (P = 0.001), diabetes (P = 0.004), stroke (P = 0.009), depression (P < 0.001), bipolar disorder (P < 0.001), and anxiety (P < 0.001), but not heart disease (P = 0.102). In older ages, the greatest loss in HRQOL was associated with incident asthma, depression, and bipolar disorder. In younger ages, the greatest loss in HRQOL was associated with arthritis (any duration) and incident diabetes and anxiety. Additionally, gender moderated the association between HRQOL and arthritis, with worse HRQOL among men with incident arthritis (P = 0.047). Loss of HRQOL associated with longer duration of chronic illness is most apparent in stroke and mental illness and differs between age groups.


A gene controlling condensation of heterochromatin in Drosophila melanogaster.
A temperature-sensitive lethal mutant of Drosophila melanogaster was used to identify an essential cell cycle function that is necessary for the mitotic condensation of heterochromatic but not of euchromatic portions of the genome. This mutant is an allele at a locus (mus-101) identified earlier by the use of mutagen-sensitive mutants. The data suggest that the mutagen-sensitive and repair-defective phenotypes of viable mus-101 mutants result from a disruption in chromosome organization.


Utility of MRI before and after uterine fibroid embolization: why to do it and what to look for.
The utility of magnetic resonance imaging (MRI) in the selection, procedure planning, and follow-up of patients undergoing arterial embolization for uterine fibroids is reviewed. Advantages of MRI over ultrasound include multiplanar imaging capability, a larger field of view, increased spatial resolution, improved anatomic detail, and the ability to detect other pelvic disorders. MRI can assess fibroid viability by detecting contrast agent enhancement. Magnetic resonance angiography has a useful role in evaluation of pelvic vasculature. Magnetic resonance parameters such as T1 and T2 relaxation times and diffusion-weighted characteristics have an emerging role in predicting outcome before and after embolization. MRI may be used to evaluate technical success and to image potential complications after embolization.


Take three, test one: a cross-sectional study to evaluate the molecular detection of Chlamydia trachomatis and Neisseria gonorrhoeae in pooled pharyngeal, anorectal and urine samples versus single-site testing among men who have sex with men in Belgium.
Objectives: To investigate the efficacy of performing a pooling strategy of triple-anatomical site samples (pharyngeal, anorectal and urine samples) for simultaneous Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) nucleic acid amplification detection.Methods: A total of 117 specimen sets (pharyngeal, anorectal and urine) were collected from 98 men between 2014 and 2016. Double sampling of pharyngeal, anorectal and urine samples allowed for pooled and unpooled analyses using a multiplex Abbott Real Time CT/NG assay, together with confirmatory PCR testing in case of CT/NG positivity. Clinical and demographic data were analyzed.Results: The positivity rate for the triple-site pooled testing for CT and NG was 8.5% (10/117) and 6.8%, (8/117), respectively, compared to the single-site testing total positivity rate, which was 9.4% (11/117) and 4.3% (5/117) for CT and NG, respectively. Pooled analysis missed one CT-positive urine sample and one CT-positive anorectal sample could not be confirmed. In addition, less PCR inhibition was reported for the pooled sample (PS) testing and ERV-3 qPCR testing revealed ineffective sampling of self-collected anorectal swabs in two cases. No pharyngeal samples were positive for CT, nor were any urine samples positive for NG.Conclusion: This small study showed that PS testing is a possible testing strategy for screening high-risk men who have sex with men attending pre-exposure prophylaxis (PrEP) clinics. However, due to the low positivity rate of CT/NG in this study, larger evaluations are needed to confirm the effectiveness of CT/NG screening with multiple-site PS nucleic acid amplification test (NAAT) screening practices.


Induced abortion as an independent risk factor for breast cancer: a comprehensive review and meta-analysis.
To ascertain, from the published reports to date, whether or not a significantly increased risk of breast cancer is specifically attributable to a history of induced abortion, independent of spontaneous abortion and age at first full term pregnancy (or first live birth); to establish the relative magnitude of such risk increase as may be found, and to ascertain and quantify such risk increases as may pertain to particular subpopulations of women exposed to induced abortion; in particular, nulliparous women and parous women exposed before compared with after the first full term pregnancy. The meta-analysis includes all 28 published reports which include specific data on induced abortion and breast cancer incidence. Since some study data are presented in more than one report, the 28 reports were determined to constitute 23 independent studies. Overall induced abortion odds ratios and odds ratios for the different subpopulations were calculated using an average weighted according to the inverse of the variance. An overall unweighted average was also computed for comparison. No quality criteria were imposed, but a narrative review of all included studies is presented for the reader's use in assessing the quality of individual studies. EXCLUDED STUDIES: All 33 published reports including data on abortion and breast cancer incidence but either pertaining only to spontaneous abortion or to abortion without specification as to whether it was induced or spontaneous. These studies are listed for the reader's information. The overall odds ratio (for any induced abortion exposure; n = 21 studies) was 1.3 (95% confidence interval of 1.2, 1.4). For comparison, the unweighted overall odds ratio was 1.4 (1.3,1.6). The odds ratio for nulliparous women was 1.3 (1.0,1.6), that for abortion before the first term pregnancy in parous women was 1.5 (1.2,1.8), and that for abortion after the first term pregnancy was 1.3 (1.1,1.5). The results support the inclusion of induced abortion among significant independent risk factors for breast cancer, regardless of parity or timing of abortion relative to the first term pregnancy. Although the increase in risk was relatively low, the high incidence of both breast cancer and induced abortion suggest a substantial impact of thousands of excess cases per year currently, and a potentially much greater impact in the next century, as the first cohort of women exposed to legal induced abortion continues to age.


Hierarchical decomposition of metabolic networks using k-modules.
The optimal solutions obtained by flux balance analysis (FBA) are typically not unique. Flux modules have recently been shown to be a very useful tool to simplify and decompose the space of FBA-optimal solutions. Since yield-maximization is sometimes not the primary objective encountered in vivo, we are also interested in understanding the space of sub-optimal solutions. Unfortunately, the flux modules are too restrictive and not suited for this task. We present a generalization, called k-module, which compensates the limited applicability of flux modules to the space of sub-optimal solutions. Intuitively, a k-module is a sub-network with low connectivity to the rest of the network. Recursive application of k-modules yields a hierarchical decomposition of the metabolic network, which is also known as branch decomposition in matroid theory. In particular, decompositions computed by existing methods, like the null-space-based approach, introduced by Poolman et al. [(2007) J. Theor. Biol. 249: , 691-705] can be interpreted as branch decompositions. With k-modules we can now compare alternative decompositions of metabolic networks to the classical sub-systems of glycolysis, tricarboxylic acid (TCA) cycle, etc. They can be used to speed up algorithmic problems [theoretically shown for elementary flux modes (EFM) enumeration] and have the potential to present computational solutions in a more intuitive way independently from the classical sub-systems.


Second malignancies as a consequence of nucleoside analog therapy for chronic lymphoid leukemias.
The nucleoside analogs fludarabine, 2'-deoxycoformycin (DCF), and 2-chlorodeoxyadenosine (CdA), commonly used in the treatment of patients with indolent lymphoid malignancies such as chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HCL), are associated with myelosuppression and profound and prolonged immunosuppression. These complications raise the possibility of an increase in secondary malignancies in patients whose disease already places them at greater risk. The purpose of the present study was to assess the frequency of second tumors in patients with CLL who are treated with fludarabine and in patients with HCL who are treated with DCF and CdA. We reviewed the long-term follow-up data for 2,014 patients treated on National Cancer Institute Group C protocols with fludarabine for relapsed and refractory CLL and with DCF and CdA for HCL using a Second Cancer Report. The numbers of observed and expected secondary tumors were compared. Median follow-up periods for the DCF (n = 409), fludarabine (n = 724), and CdA (n = 979) studies were 6.9, 7.4, and 5.1 years, respectively. The 111 malignancies were most commonly lymphoma (25 patients), prostate (19), lung (15), colorectal (nine), bladder (six), and breast (six), but also CNS, stomach, ovary, head and neck, melanoma, sarcoma, testicular, and myeloid leukemias. Compared with age-adjusted 1994 Surveillance and Epidemiology End-Results rates for the general population, the observed/expected frequencies for DCF, fludarabine, and CdA were 1.43 (95% confidence interval [CI], 0.93 to 2.10), 1.65 (95% CI, 1.04 to 2.47), and 1.50 (95% CI, 1.14 to 1.93), respectively, indicating a significant (at P =.05) increase in risk for patients treated on the latter two protocols compared with a normal population. However, these values are consistent with the increase already associated with these diseases. Despite their immunosuppression, nucleoside analogs can be safely administered to patients with CLL or HCL without a significantly increased risk of secondary malignancies.


Pulmonary non-tuberculous mycobacteria in colonisation and disease in The Gambia.
The clinical relevance of pulmonary non-tuberculous mycobacteria (PNTM) in The Gambia is unknown. The aim of this study was to estimate the prevalence of non-tuberculous mycobacteria (NTM) in colonisation, and the burden of clinically relevant pulmonary NTM (PNTM) disease in The Gambia. This was a cross-sectional study of the prevalence of NTM in participants aged ≥ 15 years, in a nationwide tuberculosis (TB) prevalence survey between December 2011 and January 2013. We enrolled 903 participants with suspected NTM and NTM cultures were confirmed by 16S rRNA gene sequencing analyses. We applied the American Thoracic Society/Infectious Disease Society of America (ATS/IDSA) diagnostic criteria to determine clinical relevance of NTM. A total of 575 participants had acid-fast bacilli (AFB) positive Mycobacterial Growth Indicator Tube (MGIT) cultures and 229 (39.8%) were NTM. M. avium complex was by far the most isolated NTM (71.0%), followed by M. fortuitum (9.5%) and M. nonchromogenicum (2.9%). Older participants (> 24 years old) were four times more likely to have NTM in their sputa. Only 20.5% (9/44) NTM cases met the ATS/IDSA criteria for NTM disease. This study provides important data on the prevalence of NTM in pulmonary samples of suspected TB cases with AFB positive cultures from a nationally representative population in The Gambia. Enhanced PNTM surveillance is recommended to better understand the contribution of NTM to pulmonary disease.


Canine post-thaw sperm quality can be predicted by using CASA, and classification and regression tree (CART)-analysis.
In individual dogs, despite good quality of raw sperm, some parameters are significantly changed after thawing, which cannot be predicted. We therefore investigated whether motility parameters objectively obtained by CASA, membrane integrity (MI), cell morphology or a combination are suitable to improve the prediction of bad post-thaw quality. For this purpose 250 sperm analysis protocols from 141 healthy stud dogs, all patients introduced for sperm cryopreservation, were evaluated and a Classification and Regression Tree (CART) -analysis performed. The sperm was routinely collected, analysed, and frozen by using a modified Uppsala system. After thawing, data were routinely examined by using CASA, fluorescent microscopy for membrane integrity (MI) and Hancock's fixation for evaluation of cell morphology. Samples were sorted by post-thaw progressive motility (P) in good (P > / = 50%, n=135) and bad freezers (P⟨50%, n=115). Among bad freezers, 73.9% showed in addition post-thaw total morphological abberations of >40% and/or MI ⟨50%. Bad freezers were significantly older than good freezers (p⟨0.05). Progressive motility (P), velocity curvilinear (VCL), mean coefficient (STR), and linear coefficient (LIN) were potential predictors for post-thaw sperm quality since specifity was best (85.8%) and sensitivity (75.4 %) and accuracy (80.4 %) good. For these objectively measured raw sperm parameters, cut-off values were calculated allowing prediction of bad post-thaw results with high accuracy: P = 83.1 % VCL = 161.3 µm/sec, STR = 0.83 %, and LIN = 0.48 %. Raw sperm samples with values below these cut off values will have below average post-thaw quality with a probability of 85.8%. We conclude that VCL, P, STR and LIN are potential predictors of the outcome of sperm cryopreservation, when combined.


Drug-induced gingival enlargement.
Side effect of medicamentous treatment in hypertension therapy and angina pectoris with calcium channel blockers related to fibrotic gingival enlargement were examined. In our study we deal with clinico-histopathological and microbiological knowledge from this field underpinned by two case reports treated with antihypertensive therapy using calcium channel blockers. In the first case report we were largely concerned with microbiological findings from the area of periodontal pseudopockets diagnosed through a DNA analysis and appropriate antibiotic therapy. In a patient treated with a preparation from amlodipine group we proceeded to a complex treatment involving the change of hypertension therapy, introduction of professional and home oral hygiene and also following surgical and prosthetico-aesthetic rehabilitation. Case Report 1 who was for a long term medicated with a preparation from the nifedipine group of antihypertensives we detected the presence of periodontal pseudopockets with probing depth of 4 to 7 mm with positive BOP and with marked rigid fibrotic gingival enlargement accompanied with considerable foetor ex ore. In a patient from the Case Report 2 who was for a long term medicated with a preparation from the amlodipine group of antihypertensives with large gingival overgrowth angiogenesis was characterized by cuboidal endothelial cell lining. In the samples under a layer of stratified epithelium there was present dense fibrous connective tissue comprising largely of collagen fiber bundles. Bacterial composition in the patient with a high degree of gingival enlargement and periodontal pseudopockets 4 to 7 mm deep represented a typical spectrum of bacteria occuring in chronic forms of periodontitis. However, we cannot determine, if such distribution of bacteria was primary before the application of nifedipine antihypertensives, or it originated later after the formation of typical anaerobic setting of false periodontal pockets.


Coffee, caffeine, and sleep: A systematic review of epidemiological studies and randomized controlled trials.
Caffeine is the most widely consumed psychoactive substance in the world. It is readily available in coffee and other foods and beverages, and is used to mitigate sleepiness, enhance performance, and treat apnea in premature infants. This review systematically explores evidence from epidemiological studies and randomized controlled trials as to whether coffee and caffeine have deleterious effects on sleep. Caffeine typically prolonged sleep latency, reduced total sleep time and sleep efficiency, and worsened perceived sleep quality. Slow-wave sleep and electroencephalographic (EEG) slow-wave activity were typically reduced, whereas stage-1, wakefulness, and arousals were increased. Dose- and timing-response relationships were established. The sleep of older adults may be more sensitive to caffeine compared to younger adults. Pronounced individual differences are also present in young people, and genetic studies isolated functional polymorphisms of genes implicated in adenosine neurotransmission and metabolism contributing to individual sensitivity to sleep disruption by caffeine. Most studies were conducted in male adults of Western countries, which limits the generalizability of the findings. Given the importance of good sleep for general health and functioning, longitudinal investigations aimed at establishing possible causal relationships among coffee- and caffeine-induced changes in sleep quality and health development are warranted.


Impaired interferon signaling is a common immune defect in human cancer.
Immune dysfunction develops in patients with many cancer types and may contribute to tumor progression and failure of immunotherapy. Mechanisms underlying cancer-associated immune dysfunction are not fully understood. Efficient IFN signaling is critical to lymphocyte function; animals rendered deficient in IFN signaling develop cancer at higher rates. We hypothesized that altered IFN signaling may be a key mechanism of immune dysfunction common to cancer. To address this, we assessed the functional responses to IFN in peripheral blood lymphocytes from patients with 3 major cancers: breast cancer, melanoma, and gastrointestinal cancer. Type-I IFN (IFN-alpha)-induced signaling was reduced in T cells and B cells from all 3 cancer-patient groups compared to healthy controls. Type-II IFN (IFN-gamma)-induced signaling was reduced in B cells from all 3 cancer patient groups, but not in T cells or natural killer cells. Impaired-IFN signaling was equally evident in stage II, III, and IV breast cancer patients, and downstream functional defects in T cell activation were identified. Taken together, these findings indicate that defects in lymphocyte IFN signaling arise in patients with breast cancer, melanoma, and gastrointestinal cancer, and these defects may represent a common cancer-associated mechanism of immune dysfunction.


Sleep Duration and Cardiovascular Health in a Representative Community Population (from NHANES, 2005 to 2016).
The 2016 American Heart Association Scientific Statement on sleep duration and cardiovascular risk suggested that optimal sleep duration is critical for cardiovascular health, with both long and short sleep duration associated with adverse health outcomes. We examined the relation between sleep duration and cardiovascular health among the general population in the United States from 2005 to 2016. We sought to investigate associations between sleep duration and the prevalence of coronary artery disease, heart failure (HF), stroke, hypertension, diabetes mellitus (DM), and hyperlipidemia. Using the National Health and Nutrition Examination Survey, we identified all patients with HF, coronary artery disease, hypertension, hyperlipidemia, DM, and stroke from 2005 to 2016. Multivariable logistic regression analyses were performed to adjust for age, sex, body mass index (BMI), marital status, educational level, physical activity, sedentary activity, depression, blood pressure, lipid profiles, and hemoglobin. In total, 32,152 National Health and Nutrition Examination Survey participants responded to the sleep survey. Both short sleepers (<7 hours, n = 12,027) and long sleepers (>9 hours, n = 1,058) were older and more likely to have a higher BMI than optimal sleepers (7 to 9 hours, n = 19,067; all p values <0.05). After adjusting for confounding variables and in comparison to those with optimal sleep duration, short sleep duration was associated with a higher prevalence of previous stroke (odds ratio [OR] 1.45; 95% confidence intervals [CI] 1.23 to 1.70), HF (OR 1.65; 95% CI 1.40 to 1.95), DM (OR 1.35; 95% CI 1.23 to 1.49), and hyperlipidemia (OR 1.12; 95% CI 1.04 to 1.22), whereas long sleep duration was associated with a higher prevalence of previous stroke (OR 1.81; 95% CI 1.37 to 2.34) and HF (OR 1.47; 95% CI 1.08 to 1.97). In conclusion, both long and short sleep durations were associated with poor cardiovascular health in this cross-sectional study.


The Second Triennial Systematic Literature Review of European Nursing Research: Impact on Patient Outcomes and Implications for Evidence-Based Practice.
European research in nursing has been criticized as overwhelmingly descriptive, wasteful and with little relevance to clinical practice. This second triennial review follows our previous review of articles published in 2010, to determine whether the situation has changed. To identify, appraise, and synthesize reports of European nursing research published during 2013 in the top 20 nursing research journals. Systematic review with descriptive results synthesis. We identified 2,220 reports, of which 254, from 19 European countries, were eligible for analysis; 215 (84.7%) were primary research, 36 (14.2%) secondary research, and three (1.2%) mixed primary and secondary. Forty-eight (18.9%) of studies were experimental: 24 (9.4%) randomized controlled trials, 11 (4.3%) experiments without randomization, and 13 (5.1%) experiments without control group. A total of 106 (41.7%) articles were observational: 85 (33.5%) qualitative research. The majority (158; 62.2%) were from outpatient and secondary care hospital settings. One hundred and sixty-five (65.0%) articles reported nursing intervention studies: 77 (30.3%) independent interventions, 77 (30.3%) interdependent, and 11 (4.3%) dependent. This represents a slight increase in experimental studies compared with our previous review (18.9% vs. 11.7%). The quality of reporting remained very poor. European research in nursing remains overwhelmingly descriptive. We call on nursing researchers globally to raise the level of evidence and, therefore, the quality of care and patient outcomes. We urge them to replicate our study in their regions, diagnose reasons for the lack of appropriate research, identify solutions, and implement a deliberate, targeted, and systematic global effort to increase the number of experimental, high quality, and relevant studies into nursing interventions. We also call on journal editors to mandate an improvement in the standards of research reporting in nursing journals.


Syntheses of C-peptides and human proinsulin.
Syntheses of human, dog, rat, and duck C-peptides and their analogues and preliminary results on the total synthesis of human proinsulin are described. In the syntheses of the C-peptides, chain elongation was performed exclusively by the azide-fragment condensation method in solution. The synthetic human, dog, rat, and duck C-peptides and their analogues were proved to be homogeneous by several analytic means. With these synthetic peptides, radioimmunoassay systems for dog, rat, and duck C-peptides were developed. For the total synthesis of human proinsulin, 10 protected peptide hydrazides were prepared, and the linearly protected hexaoctacontapeptide having the proposed sequence of human proinsulin was constructed by the azide-fragment condensation method in solution starting from the C-terminal undecapeptide (HP 75-86). After deblocking of the alpha-amino protection, the partially protected hexaoctacontapeptide was treated with sodium in liquid ammonia. The ensuing sulfhydryl form was converted to the S-sulfonate form, which was reduced and then air-oxidized. The oxidized material was purified by gel filtration on Sephadex G-50 (fine) followed by ion-exchange chromatography on DEAE-cellulose. The cross-reactivity in the insulin radioimmunoassay of the ensuing product was 62.5 per cent of porcine proinsulin on a weight basis at B/Bo = 60 per cent. Acid hydrolysis and amino acid analysis of this product gave the theoretically expected ratios. In addition, this peptide, as well as the S-sulfonate form of the hexaoctacontapeptide, showed displacement curves superimposable on that of synthetic human C-peptide on an equimolar basis in the human C-peptide radioimmunoassay (antiserum 527). These results confirm the synthesis of human proinsulin.


Neural autoantibodies and neurophysiologic abnormalities in patients exposed to molds in water-damaged buildings.
Adverse health effects of fungal bioaerosols on occupants of water-damaged homes and other buildings have been reported. Recently, it has been suggested that mold exposure causes neurological injury. The authors investigated neurological antibodies and neurophysiological abnormalities in patients exposed to molds at home who developed symptoms of peripheral neuropathy (i.e., numbness, tingling, tremors, and muscle weakness in the extremities). Serum samples were collected and analyzed with the enzyme-linked immunosorbent assay (ELISA) technique for antibodies to myelin basic protein, myelin-associated glycoprotein, ganglioside GM1, sulfatide, myelin oligodendrocyte glycoprotein, alpha-B-crystallin, chondroitin sulfate, tubulin, and neurofilament. Antibodies to molds and mycotoxins were also determined with ELISA, as reported previously. Neurophysiologic evaluations for latency, amplitude, and velocity were performed on 4 motor nerves (median, ulnar, peroneal, and tibial), and for latency and amplitude on 3 sensory nerves (median, ulnar, and sural). Patients with documented, measured exposure to molds had elevated titers of antibodies (immunoglobulin [Ig]A, IgM, and IgG) to neural-specific antigens. Nerve conduction studies revealed 4 patient groupings: (1) mixed sensory-motor polyneuropathy (n = 55, abnormal), (2) motor neuropathy (n = 17, abnormal), (3) sensory neuropathy (n = 27, abnormal), and (4) those with symptoms but no neurophysiological abnormalities (n = 20, normal controls). All groups showed significantly increased autoantibody titers for all isotypes (IgA, IgM, and IgG) of antibodies to neural antigens when compared with 500 healthy controls. Groups 1 through 3 also exhibited abnormal neurophysiologic findings. The authors concluded that exposure to molds in water-damaged buildings increased the risk for development of neural autoantibodies, peripheral neuropathy, and neurophysiologic abnormalities in exposed individuals.


Organometallic chemistry of ethynyl boronic acid MIDA ester, HC≡CB(O2CCH2)2NMe.
The reactions of HC≡CBMIDA (BMIDA = B(O2CCH2)2NMe) with a range of ruthenium complexes afford the first isolated examples of σ-alkynyl, σ-alkenyl and vinylidene complexes bearing 4-coordinate boron substituents. Specifically, the reactions of HC≡CBMIDA with [RuH(S2CNR2)(CO)(PPh3)2] (R = Me, Et) and [Ru(CO)2(PPh3)3] afford the alkynyl complexes [Ru(C≡CBMIDA)(S2CNR2)(CO)(PPh3)2] and [RuH(C≡CBMIDA)(CO)2(PPh3)2], the latter being converted to [Ru(C≡CBMIDA)Cl(CO)2(PPh3)2] on treatment with chloroform. With [RuCl(dppe)2]PF6 the vinylidene salt [RuCl(=C=CHBMIDA)(dppe)2]PF6 is obtained, which reacts with Et3N to afford the neutral alkynyl derivative [Ru(C≡CBMIDA)Cl(dppe)2]. Hydrometallation of HC≡CBMIDA by [RuHCl(CO)(PPh3)3] affords the coordinatively unsaturated σ-alkenyl complex [RuCl(CH=CHBMIDA)(CO)(PPh3)2] which in turn reacts with CO, CNC6H2Me3-2,4,6, [Et2NH2][S2CNEt2] or K[HB(pz)3] (pz = pyrazol-1-yl) to afford the coordinatively saturated complexes [Ru(CH=CHBMIDA)Cl(CO)2(PPh3)2], [Ru(CH=CHBMIDA)Cl(CO)(CNC6H2Me3)(PPh3)2], [Ru(CH=CHBMIDA)(S2CNEt2)(CO)-(PPh3)2] and [Ru(CH=CHBMIDA)(CO)(PPh3){HB(pz)3}]. In all cases, the transannular N→B dative bond is retained in the BMIDA substituent.


Prospective evaluation of access incision position for minimally invasive surgical organ exposure in cats.
To evaluate the exposure obtained for minimally invasive abdominal organ biopsy (MIOB) from 3 access incisions in cats. Prospective experimental study and clinical case series. 6 purpose-bred research cats and 6 feline clinical patients with indications for abdominal organ biopsy. Three 3-cm incisions into the peritoneal cavity were created at different locations along the linea alba in research cats in randomized order. A wound retraction device was inserted in each incision. Ability to exteriorize various abdominal organs to the extent required to reasonably perform a surgical biopsy was recorded, and results were compared among incision sites. On the basis of results obtained, the access incision that provided exposure of the most frequently biopsied abdominal organs was used to perform MIOB in 6 feline clinical patients with various underlying pathological conditions. On the basis of experiments with research cats, a 3-cm access incision centered midway between the caudal margin of the xiphoid cartilage and the umbilicus was found to provide access for MIOB for most organs. In 5 of 6 clinical patients, all of the organs of interest were biopsied successfully via this incision location, although access to all hepatic lobes and all parts of the pancreas was inconsistent. In 1 cat, conversion to an open approach was performed because a palpable mass was detected in the area of the duodenocolic ligament. Optimization of access incision location for MIOB allowed biopsy specimen collection from organs of interest to be performed in a minimally invasive manner in cats.


Diverse novel phleboviruses in sandflies from the Panama Canal area, Central Panama.
The genus Phlebovirus (order Bunyavirales, family Phenuiviridae) comprises 57 viruses that are grouped into nine species-complexes. Sandfly-transmitted phleboviruses are found in Europe, Africa and the Americas and are responsible for febrile illness and infections of the nervous system in humans. The aim of this study was to assess the genetic diversity of sandfly-transmitted phleboviruses in connected and isolated forest habitats throughout the Panama Canal area in Central Panama. In total, we collected 13 807 sandflies comprising eight phlebotomine species. We detected several strains pertaining to five previously unknown viruses showing maximum pairwise identities of 45-78 % to the RNA-dependent RNA polymerase genes of phleboviruses. Entire coding regions were directly sequenced from infected sandflies as virus isolation in cell culture was not successful. The viruses were tentatively named La Gloria virus (LAGV), Mona Grita virus (MOGV), Peña Blanca virus (PEBV), Tico virus (TICV) and Tres Almendras virus (TRAV). Inferred phylogenies and p-distance-based analyses revealed that PEBV groups with the Bujaru phlebovirus species-complex, TRAV with the Candiru phlebovirus species-complex and MOGV belongs to the proposed Icoarci phlebovirus species-complex, whereas LAGV and TICV seem to be distant members of the Bujaru phlebovirus species-complex. No specific vector or habitat association was found for any of the five viruses. Relative abundance of sandflies was similar over habitat types. Our study shows that blood-feeding insects originating from remote and biodiverse habitats harbour multiple previously unknown phleboviruses. These viruses should be included in future surveillance studies to assess their geographic distribution and to elucidate if these viruses cause symptoms of disease in animals or humans.


Clinical Integration of Quantitative Susceptibility Mapping Magnetic Resonance Imaging into Neurosurgical Practice.
To introduce quantitative susceptibility mapping (QSM), a novel magnetic resonance imaging sequence, to the field of neurosurgery. QSM is introduced both in its historical context and by providing a brief overview of the physics behind the technique tailored to a neurosurgical audience. Its application to clinical neurosurgery is then highlighted using case examples. QSM offers a quantitative assessment of susceptibility (previously considered as an artifact) via a single, straightforward gradient echo acquisition. QSM differs from standard susceptibility weighted imaging in its ability to both quantify and precisely localize susceptibility effects. Clinical applications of QSM are wide reaching and include precise localization of the deep nuclei for deep brain stimulation electrode placement, differentiation between blood products and calcification within brain lesions, and enhanced sensitivity of cerebral micrometastasis identification. We present this diverse range of QSM's clinical applications to neurosurgical care via case examples. QSM can be obtained in all patients able to undergo magnetic resonance imaging and is easily integratable into busy neuroradiology programs because of its short acquisition time and straightforward, automated offline postprocessing workflow. Clinical integration of QSM may help clinicians better identify and characterize neurosurgical lesions, thereby improving patient care.


Cow's Milk Allergy among Children with Gastroesophageal Reflux Disease.
Gastroesophageal reflux disease (GERD) and cow's milk allergy (CMA) are two common conditions that occur in infancy. This study was performed to investigate the frequency of CMA in a group of patients with GERD. Eighty-one children with signs and symptoms of GERD were enrolled in this study. All subjects received omeprazole for 4 weeks after the initial evaluation. Empirical elimination of cow's milk from the diet was started for the patients who did not respond to the omeprazole treatment. Seventy-two cases presented with gastrointestinal signs and symptoms, whereas the remaining nine cases presented with respiratory complaints. After the initial treatment with omeprazole, two thirds of the cases (54 patients, 66.7%) responded well, and all of their symptoms were resolved. Cow's milk was eliminated from the diets of the remaining 27 patients. All signs and symptoms of GERD were resolved in this group after a 4 week elimination of cow's milk from the diet. A diagnosis of CMA was considered in one third of the pediatric cases with signs and symptoms of GERD. This finding shows that CMA can mimic or aggravate all signs and symptoms of severe GERD during infancy.


Positive association and future perspectives of mitochondrial DNA copy number and telomere length - a pilot twin study.
Recent experimental and population studies have highlighted the existence of telomere-mitochondria interplay. Besides studies revealing the molecular mechanisms underlying the associations of telomere defects and mitochondrial functions, investigations of mitochondrial DNA copy number (mtDNAcn) and telomere length (TL) in healthy and disease phenotypes have likewise begun, with the aim of gaining more insights about their relationship in humans. A total of 142 asymptomatic adult twins, comprising 96 monozygotic (MZ) and 46 dizygotic (DZ) twins (mean age: 50.54 ±15.43 years), members of the Hungarian Twin Registry, were included in the analysis. Applying the qPCR standard curve method, we investigated the relationship of mtDNA copy number, telomere length and clinical data, besides assessing co-twin similarities of MZ and DZ twins for their mtDNAcn and TL measures. We found that twins were similar in their intraclass correlation coefficients irrespective of zygosity, suggesting a possibly more important role of common (shared) environmental factors compared to non-shared (unique) environmental and to a smaller degree also individual genetic influences. We confirmed a significant positive association between mtDNAcn and TL (r = 0.28, p < 0.01) in age- and sex-corrected analysis. Following bivariate estimates and correction with significant predictors, the independent positive associations were further verified. Our results extend the until now modest number of studies investigating mtDNAcn and TL simultaneously in humans. In addition, we are the first to examine the relationship between mtDNAcn and telomere length in MZ and DZ twin subjects.


Classifying individuals at high-risk for psychosis based on functional brain activity during working memory processing.
The psychosis high-risk state is accompanied by alterations in functional brain activity during working memory processing. We used binary automatic pattern-classification to discriminate between the at-risk mental state (ARMS), first episode psychosis (FEP) and healthy controls (HCs) based on n-back WM-induced brain activity. Linear support vector machines and leave-one-out-cross-validation were applied to fMRI data of matched ARMS, FEP and HC (19 subjects/group). The HC and ARMS were correctly classified, with an accuracy of 76.2% (sensitivity 89.5%, specificity 63.2%, p = 0.01) using a verbal working memory network mask. Only 50% and 47.4% of individuals were classified correctly for HC vs. FEP (p = 0.46) or ARMS vs. FEP (p = 0.62), respectively. Without mask, accuracy was 65.8% for HC vs. ARMS (p = 0.03) and 65.8% for HC vs. FEP (p = 0.0047), and 57.9% for ARMS vs. FEP (p = 0.18). Regions in the medial frontal, paracingulate, cingulate, inferior frontal and superior frontal gyri, inferior and superior parietal lobules, and precuneus were particularly important for group separation. These results suggest that FEP and HC or FEP and ARMS cannot be accurately separated in small samples under these conditions. However, ARMS can be identified with very high sensitivity in comparison to HC. This might aid classification and help to predict transition in the ARMS.


Red light cameras revisited. Recent evidence on red light camera safety effects.
The aim of this paper is to update the most recent published evidence on road safety estimates of recent red light camera (RLC) and speed/red light camera studies (SRLC). A literature search was carried out on RLC studies in the period 2013-2017 which, after screening, identified 18 recent studies on red light cameras (RLCs) and speed/red light cameras (SRLCs). The methodology and results of these studies were further examined and summary safety estimates were derived and compared to earlier meta-analysis summary estimates. The primary conclusion is that the new safety estimates from this paper show a 12% decrease of total crashes and confirm the general tendency of RLCs to reduce right angle crashes while at the same time increasing rear-end crashes. However, comparing the developments over time, the review reveals that safety estimates tend to increase with time (the safety effects of the most recent studies are the greatest). Also safety estimates suggest more positive effects with SRLCs than with RLCs. Most of recent studies indicate that spillover effects are present and can be quite substantial.


Cardiopulmonary Resuscitation Training in Schools: A Comparison of Trainee Satisfaction among Different Age Groups.
Cardiopulmonary resuscitation (CPR) has recently been added to the school curriculum worldwide and is currently taught to students between the ages of 10 and 16 years. The effect of the age of trainees on their satisfaction with CPR training has yet been elucidated. The aim of this study was to compare the satisfaction of trainees of different ages who participated in CPR training in schools in Japan. In total, 392 primary school students (10-11 years old), 1798 junior high school students (12-13 years old), and 4162 high schools students (15-16 years old) underwent the same 3-h course of CPR training, according to the guidelines of 2000 for Emergency Cardiovascular Care and CPR. The course was evaluated by a questionnaire completed by the participants. Primary school students responded most positively to all questions, including those reflecting enjoyment and the confidence of participants to apply CPR (Jonckheere-Terpstra test: P < 0.01). Exploratory factor analysis defined three latent variables (reaction, concentration, and naïveté) based on the seven variables addressed in the questionnaire. In the causal relationships analyzed by structural equation modeling (SEM), naïveté (which is related to age) directly affected the other latent variables. The current model suggested that the students' satisfaction with CPR training was strongly related to their age. Primary school students enjoyed CPR training more and were more confident in their ability to perform CPR than junior high and high school students were. Therefore, children aged 10-11 years may be the most appropriate candidates for the introduction of CPR training in schools.


How to troubleshoot the electroanatomic map.
An electroanatomical mapping system is a useful tool for complex arrhythmia ablation. The system reconstructs the precise 3-dimensional chamber of interest with electrical and anatomical information. There are several technical aspects that physicians should be aware of to maximize its efficacy. This review provides relevant information on troubleshooting of the mapping system.


Method-of-moments formulation for the analysis of plasmonic nano-optical antennas.
We present a surface integral equation (SIE) to model the electromagnetic behavior of metallic objects at optical frequencies. The electric and magnetic current combined field integral equation considering both tangential and normal equations is applied. The SIE is solved by using a method-of-moments (MoM) formulation. The SIE-MoM approach is applied only on the material boundary surfaces and interfaces, avoiding the cumbersome volumetric discretization of the objects and the surrounding space required in differential-equation formulations. Some canonical examples have been analyzed, and the results have been compared with analytical reference solutions in order to prove the accuracy of the proposed method. Finally, two plasmonic Yagi-Uda nanoantennas have been analyzed, illustrating the applicability of the method to the solution of real plasmonic problems.


Hydrogen rich saline reduces immune-mediated brain injury in rats with acute carbon monoxide poisoning.
This experiment was designed to determine whether hydrogen (H(2)) rich saline can ameliorate brain abnormalities in a rat model with acute carbon monoxide (CO) poisoning. Sprague-Dawley male rats were used for CO poisoning and H(2) rich saline treatment. Changes in neurons, microglias, and myelin sheath were observed by electron microscope. Neuron loss was assessed by Nissl staining. Antioxidant capacities were evaluated by studying superoxide dismutase activities and malondialdehyde concentration in the brain and serum. Infiltration of macrophages, expression of immune-associated cytokines (MIP-1-alpha and ICAM-1), and changes in myelin basic protein (MBP) were monitored by immunohistochemical staining and western blotting. CO-exposed rats showed the increase in neuron loss and the decrease in antioxidant capacities. And H(2) rich saline given after CO poisoning can prevent the alterations mentioned above. CO-mediated oxidative stress caused alterations in MBP, which initiated an adaptive immunological response that led to brain injury. MBP from H(2) rich saline-treated, CO-exposed rats was recognized normally by immunohistochemical staining and western blotting. Electron microscope observation from CO-exposed rats showed an apparent aggregation of microglias. Macrophages from CO-exposed rats were significantly more than those from H(2) rich saline-treated and control rats, and the immunofluorescence observation showed that macrophages were similar to microglias in type. Expression levels of MIP-1-alpha and ICAM-1 increased in the brains of CO-poisoned rats and H(2) rich saline treatment decreased the levels. The results indicate that H(2) rich saline prevents immune-mediated brain injury after CO poisoning.


Role of Krüppel-like Factor 4-p21CIP1 Axis in Breast Cancer Stem-like Cell Inhibition by Benzyl Isothiocyanate.
Cancer chemoprevention by benzyl isothiocyanate (BITC), which is derived from cruciferous vegetables like garden cress, in a transgenic mouse model of breast cancer is associated with inhibition of breast cancer stem-like cells (bCSC), but the molecular regulators of this effect remain elusive. This study demonstrates a protective effect of Krüppel-like factor 4 (KLF4)-p21CIP1 axis in bCSC inhibition by BITC. Exposure of human breast cancer cells (MCF-7, MDA-MB-231, and SUM159) to plasma-achievable concentrations of BITC resulted in a robust induction of KLF4 mRNA and its protein expression as determined by qRT-PCR and Western blotting or confocal microscopy. BITC-mediated suppression of bCSC markers, including aldehyde dehydrogenase 1 activity and mammosphere frequency, was significantly augmented by transient or stable knockdown of KLF4. Western blotting and IHC revealed relatively higher levels of KLF4 protein in mammary tumor sections from BITC-treated mice in comparison with controls, but the difference was insignificant. Analysis of the breast cancer RNA-Seq data from The Cancer Genome Atlas indicated significant positive correlation between expression of KLF4 and that of p21CIP1 (CDKN1A) but not β-Catenin (CTNNB1). Knockdown of p21CIP1 protein also amplified BITC-mediated suppression of bCSC. Finally, KLF4 was recruited to the promoter of p21CIP1 as indicated by chromatin immunoprecipitation assay. These results indicate that induction of KLF4-p21CIP1 axis attenuates inhibitory effect of BITC on bCSC self-renewal. Translational implication of these findings is that breast cancer chemoprevention by BITC may be augmented with a combination regimen involving BITC and an inhibitor of KLF4.


Cyclin/PCNA is the auxiliary protein of DNA polymerase-delta.
Identification of the cellular proteins whose expression is regulated during the cell cycle in normal cells is essential for understanding the mechanisms involved in the control of cell proliferation. A nuclear protein called cyclin of relative molecular mass 36,000 (Mr 36K), whose synthesis correlates with the proliferative state of the cell, has been identified in several cell types of human, mouse, hamster and avian origin. The rate of cyclin synthesis is very low in quiescent cells and increases several fold after serum stimulation shortly before DNA synthesis. Immunofluorescence and autoradiography studies have shown that the nuclear staining patterns of cyclin during S phase have a sequential order of appearance and a clear correlation can be found between DNA synthesis and cyclin positive nuclei. The proliferating cell nuclear antigen (PCNA) and cyclin have many common properties and it has been shown that these two are identical. Recently a protein which is required by DNA polymerase-delta for its catalytic activity with templates having low primer/template ratios has been isolated from calf thymus. We report here that cyclin and the auxiliary protein of DNA polymerase-delta are identical.


Generation and Classification of Activity Sequences for Spatiotemporal Modeling of Human Populations.
Human activity encompasses a series of complex spatiotemporal processes that are difficult to model but represent an essential component of human exposure assessment. A significant empirical data source, like the American Time Use Survey (ATUS), can be leveraged to model human activity. However, tractable models require a better stratification of activity data to inform about different, but classifiable groups of individuals, that exhibit similar activity sequences and mobility patterns. Using machine learning algorithms, we developed an unsupervised classification and sequence generation method that is capable of generating coherent and stochastic sequences of activity from the ATUS data. This classification, when combined with any spatiotemporal exposure profile, allows the development of stochastic models of exposure patterns and records for groups of individuals exhibiting similar activity behaviors.


Cellular Immunotherapy and Locoregional Administration of CAR T-Cells in Malignant Pleural Mesothelioma.
Malignant pleural mesothelioma (MPM) is a treatment recalcitrant tumor with a poor overall survival (OS). Current approved treatment consists of first line chemotherapy that only modestly increases OS, illustrating the desperate need for other treatment options in MPM. Unfortunately, clinical studies that investigate the effectivity of checkpoint inhibitor (CI) treatment failed to improve clinical outcome over current applied therapies. In general, MPM is characterized as an immunological cold tumor with low T-cell infiltration, which could explain the disappointing results of clinical trials investigating CI treatment in MPM. Currently, many other therapeutic approaches, such as cellular therapies and cancer vaccines are investigated that could induce a tumor-specific immune response and increase of the number of tumor-infiltrating lymphocytes. In this review we will discuss these novel treatment approaches for MPM.


Renal atrial peptide receptors and natriuresis in two-kidney, one clip hypertension.
It has been suggested that the impaired natriuretic response of the clipped kidney in two-kidney, one clip hypertensive rats is related to downregulation of renal atrial natriuretic peptide receptors. To test this hypothesis, blood volume expansion and atrial peptide binding studies were performed in this model. Infusion of 1% and then 1.5% body weight donor blood (n = 6) caused a progressive increase in plasma immunoreactive atrial natriuretic peptide (107 +/- 26 to 168 +/- 31 to 427 +/- 154 pg/ml, p less than 0.001); the sodium excretion of the nonclipped kidney rose from 230 to 2,200 to 4,000 neq/min (p less than 0.01) but that of the clipped kidney did not rise significantly. There was a highly significant correlation between log cyclic guanosine monophosphate and log sodium excretion by the nonclipped (r2 = 0.749) but not the clipped (r2 = 0.046) kidney. Between clipped and nonclipped kidneys, the association constant (5.26 +/- 0.89 versus 5.17 +/- 0.64 x 10(9)/mol) and apparent binding site density (575 +/- 92 versus 500 +/- 74 fmol/mg protein) for atrial peptide binding in isolated glomeruli did not differ. Assay of atrial peptide-induced cyclic guanosine monophosphate release by isolated glomeruli showed that clipped and nonclipped kidneys were equally responsive. Binding affinity and receptor density did not differ in homogenates prepared from inner medullas of clipped and nonclipped kidneys. These results show that the blunted natriuretic response in clipped kidneys was not associated with any relative decrease in number or function of glomerular or papillary atrial natriuretic peptide receptors.


The white-coat response in ambulatory blood pressure monitoring: elimination and attenuation.
BACKGROUND: The white-coat response can be identifiede on ambulatory blood pressure (ABP) monitoring (ABPM) and it is seen for around 2 h if the ABPM period is started in the hospital. The white-coat response to ABPM can be calculated by subtracting the 25th h ABP from the 1st h ABP; this eliminates any effect of the 24 h variation in blood pressure. METHODS: In this report we have examined whether the white-coat response is due to the environment alone or partly due to the ABPM machine (study 1). We also examined whether the white-coat response found with ABPM attenuates on repeated visits (study 2). In study 1 we started ABPM at the subject's home 3 h after an initial explanation and measurement of blood pressure in the hospital (hospital blood pressure) and examined the white-coat response (measured as the 1st h - 25th h ABP). In study 2 we performed ABPM on three occasions starting ABPM in the hospital and examined the magnitude of the white-coat response at each visit. RESULTS: In study 1 the hospital blood pressure was significantly higher than the hourly mean blood pressure at the same time of day on the following day. There was no significant difference between the 1st and 25th hourly means or the 2nd and 26th hourly means after the monitoring had been restarted at home. In study 2, the 1st and 25th mean hourly ambulatory blood pressures showed a significant reduction in the 1st h blood pressure over the three visits whereas the 25th h blood pressure was similar. The 1st h - 25 h systolic ambulatory blood pressure was significantly reduced ( P = 0.025) whereas the reduction in diastolic ambulatory blood pressure did not attain statistical significance (P = 0.182). CONCLUSION: The ABPM device itself does not cause the white-coat response seen on ABPM. This response appears to be related to the environment in which ABPM is commenced and can be eliminated by calculating the ABP beginning 3 h after the subject has left the hospital environment. Attenuation of this response is seen on repeated ABPM.


Optimal interpretation of FDG PET in the diagnosis, staging and management of pancreatic carcinoma.
This study had two purposes: to optimize the semiquantitative interpretation of 18F-fluorodeoxyglucose (FDG) PET scans in the diagnosis of pancreatic carcinoma by analyzing different cutoff levels for the standardized uptake value (SUV), with and without correction for serum glucose level (SUV(gluc)); and to evaluate the usefulness of FDG PET when used in addition to CT for the staging and management of patients with pancreatic cancer. Sixty-five patients who presented with suspected pancreatic carcinoma underwent whole-body FDG PET in addition to CT imaging. The PET images were analyzed visually and semiquantitatively using the SUV and SUV(gluc). The final diagnosis was obtained by pathologic (n = 56) or clinical and radiologic follow-up (n = 9). The performance of CT and PET at different cutoff levels of SUV was determined, and the impact of FDG PET in addition to CT on patient management was reviewed retrospectively. Fifty-two patients had proven pancreatic carcinoma, whereas 13 had benign lesions, including chronic pancreatitis (n = 10), benign biliary stricture (n = 1), pancreatic complex cyst (n = 1) and no pancreatic pathology (n = 1). Areas under receiver operating characteristic curves were not significantly different for SUV and SUV(gluc). Using a cutoff level of 3.0 for the SUV, FDG PET had higher sensitivity and specificity than CT in correctly diagnosing pancreatic carcinoma (92% and 85% versus 65% and 61%). There were 2 false-positive PET (chronic pancreatitis, also false-positive with CT) and 4 false-negative PET (all with true-positive CT, abnormal but nondiagnostic) examinations. There were 5 false-positive CT (4 chronic pancreatitis and 1 pancreatic cyst) and 18 false-negative CT (all with true-positive FDG PET scans) examinations. FDG PET clarified indeterminate hepatic lesions or identified additional distant metastases (or both) in 7 patients compared with CT. Overall, FDG PET altered the management of 28 of 65 patients (43%). FDG PET is more accurate than CT in the detection of primary tumors and in the clarification and identification of hepatic and distant metastases. The optimal cutoff value of FDG uptake to differentiate benign from malignant pancreatic lesions was 2.0. Correction for serum glucose did not significantly improve the accuracy of FDG PET. Although FDG PET cannot replace CT in defining local tumor extension, the application of FDG PET in addition to CT alters the management in up to 43% of patients with suspected pancreatic cancer.


Is a tourniquet beneficial in total knee replacement surgery? A meta-analysis and systematic review.
Proponents of tourniquets postulate that they optimise intra-operative visibility and reduce blood loss. This study compared the outcomes of tourniquet assisted to non-tourniquet assisted total knee replacement (TKR). A systematic review was undertaken of the electronic databases Medline, CINAHL, AMED and EMBASE, in addition to a review of unpublished material and a hand search of pertinent orthopaedic journals. The evidence-base was critically appraised using a tool from the Cochrane Bone, Joint and Muscle Trauma Group. Fifteen studies were identified evaluating 16 outcome measures and parameters of 1040 TKRs in 991 patients. There was a significantly greater intra-operative blood loss in non-tourniquet compared to tourniquet assisted surgery (p=0.004). There was no significant difference between the groups for total blood loss or transfusion rate (p=0.22; p=0.48). There was a trend for greater complications in tourniquet compared to non-tourniquet patients. There was no difference between the groups for any other outcome measure assessed. In conclusion, this systematic review has found that there is no advantage to using a tourniquet in knee replacement surgery for reduction of transfusion requirements.


Glycyrrhizic acid and 18β-glycyrrhetinic acid recover glucocorticoid resistance via PI3K-induced AP1, CRE and NFAT activation.
Glucocorticoids are widely used in the clinical setting as remedies for inflammatory diseases, such as asthma and chronic obstructive pulmonary disease. However, the constant increase in the number of patients suffering from glucocorticoid resistance could present a serious problem for clinicians. In these cases, it may be reasonable to use additional treatments to restore the therapeutic effect of glucocorticoids. Glycyrrhizic acid (GA) and 18β-glycyrrhetinic acid (18βGA) are bioactive compounds in licorice that have been used for thousands of years in traditional Chinese medicine to treat coughs. We showed that GA and 18βGA exhibit potential anti-inflammatory and antioxidant properties. GA and 18βGA induced dual specificity protein phosphatase 1 (DUSP1) expression, and this effect was unchanged by the addition of RU486, a glucocorticoid receptor antagonist. The stimulation of DUSP1 expression by GA and 18βGA occurred via both glucocorticoid receptor (GR) and PI3K signaling, and the simultaneous activation of transcription elements, such as AP1 (activator protein 1), CRE (cAMP response element), GRE (glucocorticoid receptor element) and NFAT (nuclear factor of activated T-cells), was confirmed. Furthermore, we designed an in vitro glucocorticoid resistance model to verify the effects of GA and 18βGA on glucocorticoid resistance that was induced by ROS. The data showed that these two phytochemicals restored glucocorticoid sensitivity by depleting ROS through HO-1 expression. p38 and NO, which are factors that are induced by reactive oxygen species and caused depletion of GR signaling, were inhibited by GA and 18βGA treatment. This phenomenon was considered to be related to the coordinated modulation of GR and PI3K signaling by GA and 18βGA, in conjugation with AP1, CRE, GRE and NFAT activation. This study provides a possible strategy for enhancing the efficacy of glucocorticoids and may improve the prognosis of patients with serious inflammatory diseases.


Monitoring and measurement of microalgae using the first derivative of absorbance and comparison with chlorophyll extraction method.
Monitoring of microalgae in water supplies and industrial applications are becoming increasingly important, yet there are few options available that are simple and accurate, and can provide real-time information. The present work illustrates a new method to determine the concentration of microalgae in water and wastewater using spectrophotometry and the first derivative of absorbance. Chlorella vulgaris was used as an indicator microalga, spiked in water samples representing a range of water qualities (distilled water, surface water, and wastewater), and correlations among C. vulgaris concentrations, absorbance, and the first derivative of absorbance measurements were investigated. In addition, detection limits were established and sensitivity analyses were carried out to determine the lowest C. vulgaris concentrations that can be confidently measured in different water matrices. Finally, the study compared the performance and detection limits of the spectrophotometry-based methods with the well-accepted chlorophyll extraction method. A strong linear relationship (R2 > 0.97) was found between C. vulgaris concentration and absorbance at 695 nm. Using the first derivative of absorbance improved C. vulgaris detection limits by reducing the effects of the background noise and interferences from other substances. The detection limits established using the first derivative method were 0.47, 0.56, and 1.96 mg TVS/L in distilled water, surface water, and wastewater, respectively. In comparison, the detection limits of the chlorophyll extraction method were found to be 19.6, 38.6, and 48.3 mg TVS/L in the same water matrices. These results indicate that first derivative of absorbance can be successfully used for monitoring of microalgae in surface waters and environmental samples as well as in bioreactors used for microalgae cultivation in industrial applications.


Immune T lymphocyte to tumor cell adhesion. Magnesium sufficient, calcium insufficient.
The prelytic adhesion of immune cytolytic thymus-derived lymphocytes to specific antigen-bearing ascites tumor target cells has been studied. A new assay was used in which adhesions are permitted to form for 2.5 min; the cells are then dispersed to prevent further adhesion, and the predispersion adhesions are quantitated by subsequent 51Cr release from the tumor cells as a result of cytolytic activity of the adhering lymphocytes. There were the following new findings: (a) magnesium is sufficient to support optimal adhesion formation even when EGTA is added to remove contaminating traces of calcium; (b) calcium supports no adhesion formation when traces of contaminating magnesium are removed by pretreating the medium with a chelating ion exchange resin; (c) calcium synergizes with suboptimal magnesium, increasing the apparent adhesion-supporting potency of magnesium 20-fold in the presence of 50 microM calcium; (d) in the presence of optimal magnesium (2--4 mM), calcium has not effect on the properties of the adhesion by any of six criteria; and (e) manganese supports adhesion better than magnesium, and strontium is ineffective. A survey of previous literature indicates that these results are remarkably similar to the predominant pattern for nonimmunologic cell adhesion (e.g., fibroblasts) involving cells from a variety of tissues in late embryonic and adult avians and mammals. This suggests that a "magnesium sufficient, calcium insufficient" mechanism may be found among the latter types of cell adhesions when appropriately examined. Moreover, it seems that the present lymphocyte-tumor cell adhesion, although evoked by specific receptor-antigen recognition, relies predominantly on mechanisms common to nonimmunologic intercellular adhesion processes.


Magnetic nanoparticles loaded with functional RNA nanoparticles.
RNA is now widely acknowledged not only as a multifunctional biopolymer but also as a dynamic material for constructing nanostructures with various biological functions. Programmable RNA nanoparticles (NPs) allow precise control over their formulation and activation of multiple functionalities, with the potential to self-assemble in biological systems. These attributes make them attractive for drug delivery and therapeutic applications. In the present study, we demonstrate the ability of iron oxide magnetic nanoparticles (MNPs) to deliver different types of RNA NPs functionalized with dicer substrate RNAs inside human cells. Our results show that use of functionalized RNA NPs result in statistically higher transfection efficiency compared to the use of RNA duplexes. Furthermore, we show that the nucleic acids in the MNP/RNA NP complexes are protected from nuclease degradation and that they can achieve knockdown of target protein expression, which is amplified by magnetic stimulus. The current work represents the very first report indicating that iron oxide nanoparticles may efficiently protect and deliver programmable RNA NPs to human cells.


Performance of a veterinary urine dipstick paddle system for diagnosis and identification of urinary tract infections in dogs and cats.
To evaluate the performance of a veterinary urine dipstick paddle (UDP) for diagnosis and identification of urinary tract infection (UTI) in dogs and cats. Prospective, randomized, blinded study. 207 urine specimens. UDPs were inoculated by 2 investigators and incubated according to manufacturer's instructions. Results, including presence or absence of bacterial growth, organism counts, and identification of uropathogens, were compared between investigators and with microbiology laboratory results. A subset of UDPs with bacterial growth was submitted to the laboratory for confirmation. The laboratory reported 64 (30.9%) specimens had growth of bacteria. Bacterial growth was reported for 63 (30.4%) and 58 (28.0%) of the UDPs by investigators 1 and 2, respectively. Sensitivity and specificity of the UDP for detection of bacterial growth were 97.3% and 98.6%, respectively, for investigator 1 and 89.1% and 99.3%, respectively, for investigator 2. For UPDs with ≥ 10(5) colony-forming units/mL, organism counts correlated well between the laboratory and investigators 1 (r = 0.95) and 2 (r = 0.89). Pathogen identification was not always accurate. Only 25 of 33 (75.8%) UDPs submitted for confirmation yielded bacteria consistent with those isolated from the original bacterial culture of urine. The veterinary UDP system was a sensitive test for screening patients for bacterial UTI, but uropathogen identification was not always accurate. When UDPs have bacterial growth, a fresh urine specimen should be submitted to the laboratory to confirm the identity of the organisms and to permit antimicrobial susceptibility testing.


Tracking the global spread of vaccine sentiments: the global response to Japan's suspension of its HPV vaccine recommendation.
In June 2013 the Japanese Ministry of Health, Labor, and Welfare (MHLW) suspended its HPV vaccination recommendation after a series of highly publicized alleged adverse events following immunization stoked public doubts about the vaccine's safety. This paper examines the global spread of the news of Japan's HPV vaccine suspension through online media, and takes a retrospective look at non-Japanese media sources that were used to support those claiming HPV vaccine injury in Japan. Two searches were conducted. One searched relevant content in an archive of Google Alerts on vaccines and vaccine preventable diseases. The second search was conducted using Google Search on January 6th 2014 and on July 18th 2014, using the keywords, "HPV vaccine Japan" and "cervical cancer vaccine Japan." Both searches were used as Google Searches render more (and some different) results than Google Alerts. Online media collected and analyzed totalled 57. Sixty 3 percent were published in the USA, 23% in Japan, 5% in the UK, 2% in France, 2% in Switzerland, 2% in the Philippines, 2% in Kenya and 2% in Denmark. The majority took a negative view of the HPV vaccine, the primary concern being vaccine safety. The news of Japan's suspension of the HPV vaccine recommendation has traveled globally through online media and social media networks, being applauded by anti-vaccination groups but not by the global scientific community. The longer the uncertainty around the Japanese HPV vaccine recommendation persists, the further the public concerns are likely to travel.


Changes in resting state functional connectivity after repetitive transcranial direct current stimulation applied to motor cortex in fibromyalgia patients.
Fibromyalgia (FM) is a chronic, centralized pain condition characterized by alterations in the functional, chemical, and structural brain networks responsible for sensory and mood processing. Transcranial direct current stimulation (tDCS) has emerged as a potential treatment for FM. tDCS can alter functional connectivity (FC) in brain regions underneath and distant to the stimulating electrode, although the analgesic mechanisms of repetitive tDCS remain unknown. The aim of this study was to investigate how a clinically relevant schedule of tDCS sessions alters resting state FC and how these changes might relate to clinical pain. Resting state functional magnetic resonance imaging data were collected from 12 patients with FM at baseline, after 5 days of sham treatment, and after 5 days of real tDCS with the anode over the left primary motor cortex (M1) and the cathode over the right supraorbital cortex. Seed to whole-brain FC analyses were performed with seed regions placed in bilateral M1, primary somatosensory cortices (S1), ventral lateral (VL) and ventral posterolateral (VPL) thalami, and periaqueductal gray (PAG). Stronger baseline FC between M1-VL thalamus, S1-anterior insula, and VL thalamus-PAG predicted greater analgesia after sham and real tDCS. Sham treatment (compared with baseline) reduced FC between the VPL thalamus, S1, and the amygdala. Real tDCS (compared with sham treatment) reduced FC between the VL thalamus, medial prefrontal, and supplementary motor cortices. Interestingly, decreased FC between the VL/VPL thalamus and posterior insula, M1, and S1 correlated with reductions in clinical pain after both sham and active treatments. These results suggest that while there may be a placebo response common to both sham and real tDCS, repetitive M1 tDCS causes distinct changes in FC that last beyond the stimulation period and may produce analgesia by altering thalamic connectivity.


Society of Toxicologic Pathology position paper on pathology image data: compliance with 21 CFR Parts 58 and 11.
The Society of Toxicologic Pathology (STP) has developed the following recommendations for the use of pathology images in compliance with the Code of Federal Regulations (CFR), Volume 21, Part 58 (Good Laboratory Practices [GLP]) and Part 11 (Electronic Records/Signatures). These recommendations include: (1) based on current technologies and practices, pathology images (printed, electronic, or digital) used for data generation (e.g., to make a diagnosis or for morphometric analysis) are raw data that must be authenticated and archived; (2) authentication of an image may be done either by initialing and dating a print of the image or by specifically annotating the electronic image file in compliance with Part 11 regulations; (3) images used for raw data are subject to GLP procedures and controls in order to ensure data integrity including written Standard Operating Procedures, testing/validation of equipment, training of personnel, etc.; (4) validation and/or performance qualification of imaging systems used to support GLP studies must be documented and any exceptions to full validation/qualification must be described in the GLP Compliance Statement for the study; (5) images that are not used for data generation are illustrative images, are not raw data, and generally do not have to be archived; 6) illustrative images should not be used to re-evaluate or supersede the pathologist's diagnosis.


Design, synthesis and biological evaluation of novel betulinic acid derivatives.
Tumor, is one of the major reason for human death, due to its widespread occurrence. Betulinic acid derivatives have attracted considerable attention as cancer chemopreventive agents and also as cancer therapeutics. Many of its derivatives inhibit the growth of human cancer cell lines by triggering apoptosis. With this background, we planned to synthesize a series of betulinic acid derivatives to assess their antiproliferation efficacy on human cancer cell lines. A series of novel betulinic acid derivatives were designed and synthesized as highlighted by the preliminary antitumor evaluation against MGC-803, PC3, A375, Bcap-37 and A431 human cancer cell lines in vitro. The pharmacological results showed that some of the compounds displayed moderate to high levels of antitumor activities with most of new exhibiting higher inhibitory activities compared to BA. The IC50 values of compound 3c on the five cancer cell lines were 2.3, 4.6, 3.3, 3.6, and 4.3 μM, respectively. Subsequent fluorescence staining and flow cytometry analysis (FCM) indicated that compound 3c could induce apoptosis in MGC-803 and PC3 cell lines, and the apoptosis ratios reached the peak (37.38% and 33.74%) after 36 h of treatment at 10 μM. This study suggests that most of betulinic acid derivatives could inhibit the growth of human cancer cell lines. Furthermore, compound 3c could induce apoptosis of cancer cells.


Diurnal Fluctuations of Verticality Perception - Lesser Precision Immediately after Waking up in the Morning.
Internal estimates of direction of gravity are continuously updated by integrating vestibular, visual and proprioceptive input, and prior experience about upright position. Prolonged static roll-tilt biases perceived direction of gravity by adaptation of peripheral sensors and central structures. We hypothesized that in the morning after sleep, estimates of direction of gravity [assessed by the subjective visual vertical (SVV)] are less precise than in the evening because of adaptation to horizontal body position and lack of prior knowledge about upright position. Using a mobile SVV-measuring device, verticality perception was assessed in seven healthy human subjects on 7 days in the morning immediately after waking up and in the evening while sitting upright. Paired t-tests were applied to analyze diurnal changes in SVV trial-to-trial variability. Average SVV variability in the morning was significantly larger than in the evening (1.9 ± 0.6° vs. 0.9 ± 0.3°, p = 0.002). SVV accuracy was not significantly different (-1.2 ± 0.9° vs. -0.4 ± 0.4°, morning vs. evening, p = 0.058) and was within normal range (±2.3°) in all but one subject. A good night's sleep has a profound effect on the brain's ability to estimate direction of gravity. Resulting variability was significantly worse after waking up reaching values more than twice as large as in the evening while there was no significant impact on SVV accuracy. We hypothesize that lacking prior knowledge, adaptation of peripheral sensors, and lower levels of arousal and cerebral metabolism contribute to such impoverished estimates. Our observations have considerable clinical impact as they indicate an increased risk for falls and fall-related injuries in the morning.


Up-regulation of Hsp72 and keratin16 mediates wound healing in streptozotocin diabetic rats.
Impaired wound healing is a complication of diabetes and a serious problem in clinical practice. We previously found that whey protein (WP) was able to regulate wound healing normally in streptozotocin (STZ)-diabetic models. This subsequent study was designed to assess the effect of WP on heat shock protein-72 (Hsp72) and keratin16 (Krt16) expression during wound healing in diabetic rats. WP at a dosage of 100 mg/kg of body weight was orally administered daily to wounded normal and STZ-diabetic rats for 8 days. At day 4, the WP-treated diabetic wound was significantly reduced compared to that in the corresponding control. Diabetic wounded rats developed severe inflammatory infiltration and moderate capillary dilatation and regeneration. Treated rats had mild necrotic formation, moderate infiltration, moderate to severe capillary dilatation and regeneration, in addition to moderate epidermal formation. Hsp72 and Krt16 densities showed low and dense activity in diabetic wounded and diabetic wounded treated groups, respectively. At day 8, WP-treatment of diabetic wounded animals revealed great amelioration with complete recovery and closure of the wound. Reactivity of Hsp72 and Krt16 was reversed, showing dense and low, or medium and low, activity in the diabetic wounded and diabetic wounded treated groups, respectively. Hsp72 expression in the pancreas was found to show dense reactivity with WP-treated diabetic wound rats. This data provides evidence for the potential impact of WP in the up-regulation of Hsp72 and Krt16 in T1D, resulting in an improved wound healing process in diabetic models.


Hypothalamic hamartoma: Neuropathology and epileptogenesis.
Hypothalamic hamartomas (HHs) are congenital malformations of the ventral hypothalamus resulting in treatment-resistant epilepsy and are intrinsically epileptogenic for the gelastic seizures that are the hallmark symptom of this disorder. This paper reviews the neuropathologic features of HHs associated with epilepsy, with an emphasis on characterizing neuron phenotypes and an ultimate goal of understanding the cellular model of ictogenesis occurring locally within this tissue. We also present previously unpublished findings on Golgi staining of HH. The microarchitecture of HH is relatively simple, with nodular clusters of neurons that vary in size and abundance with poorly defined boundaries. Approximately 80-90% of HH neurons have an interneuron-like phenotype with small, round soma and short, unbranched processes that lack spines. These neurons express glutamic acid decarboxylase and likely utilize γ-aminobutyric acid (GABA) as their primary neurotransmitter. They have intrinsic membrane properties that lead to spontaneous pacemaker-like firing activity. The remaining HH neurons are large cells with pleomorphic, often pyramidal, soma and dendrites that are more likely to be branched and have spines. These neurons appear to be excitatory, projection-type neurons, and have the functionally immature behavior of depolarizing and firing in response to GABA ligands. We hypothesize that the irregular neuronal clusters are the functional unit for ictogenesis. Further research to define and characterize these local networks is required to fully understand the cellular mechanisms responsible for gelastic seizures.


MECHANISMS IN ENDOCRINOLOGY: Endogenous subclinical hypercortisolism and bone: a clinical review.
In recent years, the condition of subclinical hypercortisolism (SH) has become a topic of growing interest. This is due to the fact that SH prevalence is not negligible (0.8-2% in the general population) and that, although asymptomatic, this subtle cortisol excess is not harmless, being associated with an increased risk of complications, in particular of osteoporosis and fragility fractures. As specific symptoms of hypercortisolism are absent in SH, the SH diagnosis relies only on biochemical tests and it is a challenge for physicians. As a consequence, even the indications for the evaluation of bone involvement in SH patients are debatable and guidelines are not available. Finally, the relative importance of bone density, bone quality and glucocorticoid sensitivity in SH is a recent field of research. On the other hand, SH prevalence seems to be increased in osteoporotic patients, in whom a vertebral fracture may be the presenting symptom of an otherwise asymptomatic cortisol excess. Therefore, the issue of who and how to screen for SH among the osteoporotic patients is widely debated. The present review will summarize the available data regarding the bone turnover, bone mineral density, bone quality and risk of fracture in patients with endogenous SH. In addition, the role of the individual glucocorticoid sensitivity in SH-related bone damage and the problem of diagnosing and managing the bone consequences of SH will be reviewed. Finally, the issue of suspecting and screening for SH patients with apparent primary osteoporosis will be addressed.


Insights into severe 5,10-methylenetetrahydrofolate reductase deficiency: molecular genetic and enzymatic characterization of 76 patients.
5,10-Methylenetetrahydrofolate reductase (MTHFR) deficiency is the most common inherited disorder of folate metabolism and causes severe hyperhomocysteinaemia. To better understand the relationship between mutation and function, we performed molecular genetic analysis of 76 MTHFR deficient patients, followed by extensive enzymatic characterization of fibroblasts from 72 of these. A deleterious mutation was detected on each of the 152 patient alleles, with one allele harboring two mutations. Sixty five different mutations (42 novel) were detected, including a common splicing mutation (c.1542G>A) found in 21 alleles. Using an enzyme assay in the physiological direction, we found residual activity (1.7%-42% of control) in 42 cell lines, of which 28 showed reduced affinity for nicotinamide adenine dinucleotide phosphate (NADPH), one reduced affinity for methylenetetrahydrofolate, five flavin adenine dinucleotide-responsiveness, and 24 abnormal kinetics of S-adenosylmethionine inhibition. Missense mutations causing virtually absent activity were found exclusively in the N-terminal catalytic domain, whereas missense mutations in the C-terminal regulatory domain caused decreased NADPH binding and disturbed inhibition by S-adenosylmethionine. Characterization of patients in this way provides a basis for improved diagnosis using expanded enzymatic criteria, increases understanding of the molecular basis of MTHFR dysfunction, and points to the possible role of cofactor or substrate in the treatment of patients with specific mutations.


Testing the near field/far field model performance for prediction of particulate matter emissions in a paint factory.
A Near Field/Far Field (NF/FF) model is a well-accepted tool for precautionary exposure assessment but its capability to estimate particulate matter (PM) concentrations is not well studied. The main concern is related to emission source characterization which is not as well defined for PM emitters compared to e.g. for solvents. One way to characterize PM emission source strength is by using the material dustiness index which is scaled to correspond to industrial use by using modifying factors, such as handling energy factors. In this study we investigate how well the NF/FF model predicts PM concentration levels in a paint factory. PM concentration levels were measured during big bag and small bag powder pouring. Rotating drum dustiness indices were determined for the specific powders used and applied in the NF/FF model to predict mass concentrations. Modeled process specific concentration levels were adjusted to be similar to the measured concentration levels by adjusting the handling energy factor. The handling energy factors were found to vary considerably depending on the material and process even-though they have the same values as modifying factors in the exposure models. This suggests that the PM source characteristics and process-specific handling energies should be studied in more detail to improve the model-based exposure assessment.


Distal radius nonunion after epiphyseal plate fracture in a 15 years old young rider.
Radius and ulna fractures are the most common long bone fractures in children and adolescents. The majority of these injuries involve the distal metaphyseal portion of the radius associated or not to physeal plate injuries. Because of the high remodelling potential of the distal radius in growing children most injuries heal without complication after closed reduction and immobilization in a long arm cast. Nonunions of closed distal radius fracture are an extremely rare occurrence especially in paediatric population. Methods: In this report, we describe a rare case of distal radius fracture nonunion in a 15-years old male rider treated conservatively with cast immobilization. Eight months later he underwent surgical closed reduction and fixation with kirschner wire and cannulated screw. Results: Follow-up at 2 years showed satisfying radiological and functional outcomes. The patient ultimately returned to ride 3 months following surgery. Conclusions: Nonunion is rarely seen in distal radius fractures in healthy children and adolescents, and there are few studies in the literature. Treatment of the nonunion must be individualized and the results are not entirely predictable.


Evolutionarily conserved elements in vertebrate, insect, worm, and yeast genomes.
We have conducted a comprehensive search for conserved elements in vertebrate genomes, using genome-wide multiple alignments of five vertebrate species (human, mouse, rat, chicken, and Fugu rubripes). Parallel searches have been performed with multiple alignments of four insect species (three species of Drosophila and Anopheles gambiae), two species of Caenorhabditis, and seven species of Saccharomyces. Conserved elements were identified with a computer program called phastCons, which is based on a two-state phylogenetic hidden Markov model (phylo-HMM). PhastCons works by fitting a phylo-HMM to the data by maximum likelihood, subject to constraints designed to calibrate the model across species groups, and then predicting conserved elements based on this model. The predicted elements cover roughly 3%-8% of the human genome (depending on the details of the calibration procedure) and substantially higher fractions of the more compact Drosophila melanogaster (37%-53%), Caenorhabditis elegans (18%-37%), and Saccharaomyces cerevisiae (47%-68%) genomes. From yeasts to vertebrates, in order of increasing genome size and general biological complexity, increasing fractions of conserved bases are found to lie outside of the exons of known protein-coding genes. In all groups, the most highly conserved elements (HCEs), by log-odds score, are hundreds or thousands of bases long. These elements share certain properties with ultraconserved elements, but they tend to be longer and less perfectly conserved, and they overlap genes of somewhat different functional categories. In vertebrates, HCEs are associated with the 3' UTRs of regulatory genes, stable gene deserts, and megabase-sized regions rich in moderately conserved noncoding sequences. Noncoding HCEs also show strong statistical evidence of an enrichment for RNA secondary structure.


Acetylation of the C terminus of Ku70 by CBP and PCAF controls Bax-mediated apoptosis.
Apoptosis is a key tumor suppression mechanism that can be initiated by activation of the proapoptotic factor Bax. The Ku70 DNA end-joining protein has recently been shown to suppress apoptosis by sequestering Bax from mitochondria. The mechanism by which Bax is regulated remains unknown. Here, we identify eight lysines in Ku70 that are targets for acetylation in vivo. Five of these, K539, K542, K544, K533, and K556, lie in the C-terminal linker domain of Ku70 adjacent to the Bax interaction domain. We show that CBP and PCAF efficiently acetylate K542 in vitro and associate with Ku70 in vivo. Mimicking acetylation of K539 or K542 or treating cells with deacetylase inhibitors abolishes the ability of Ku70 to suppress Bax-mediated apoptosis. We demonstrate that increased acetylation of Ku70 disrupts the Ku70-Bax interaction and coincides with cytoplasmic accumulation of CBP. These results shed light on the role of acetyltransferases as tumor suppressors.


Dog walking is associated with a favorable risk profile independent of moderate to high volume of physical activity.
An innovative strategy for helping people achieve recommended levels of daily physical activity is dog walking. We assessed differences in physical activity and risk indicators between dog owners who 1) walk their dog (n = 399) and 2) do not walk their dog (n = 137) and compared them with adults who do not own dogs (n = 380). Participants (39 ± 13 years) were recruited online and completed an electronic questionnaire. Healthy People 2010 risk indicators included physical activity, overweight status, tobacco use, nutrition behaviors, chronic conditions, depressive symptoms, and social support. Compared with dog walkers, those who did not own or walk their dog reported less physical activity (MET-min·week-1) and a higher body mass index (P < .01). Moreover, after adjusting for age and moderate to high physical activity, those who did not own dogs had significantly greater odds of self-reported diabetes [OR = 2.53; 95%CI (1.17-5.48)], hypertension [OR = 1.71; 95%CI (1.03-2.83)], hypercholesterolemia [OR = 1.72; 95%CI (1.06-2.81)], and depression [OR = 1.49; 95%CI (1.09-2.05)] compared with participants who regularly walked their dogs. Because of the health benefits associated with dog walking, this activity should be encouraged within communities as a method of promoting and sustaining a healthy lifestyle.