Genital warts and vulvar intraepithelial neoplasia: natural history and effects of treatment and human immunodeficiency virus infection.
To describe the natural history of genital warts and vulvar intraepithelial neoplasia (VIN) in women with human immunodeficiency virus (HIV). A cohort of 2,791 HIV-infected and 953 uninfected women followed for up to 13 years had genital examinations at 6-month intervals with biopsy for lesions suspicious for VIN. The prevalence of warts was 4.4% (5.3% for HIV-seropositive women and 1.9% for HIV-seronegative women, P<.001). The cumulative incidence of warts was 33% (95% confidence interval [CI] 30-36%) in HIV-seropositive and 9% (95% CI 6-12%) in HIV-seronegative women (P<.001). In multivariable analysis, lower CD4 lymphocyte count, younger age, and current smoking were strongly associated with risk for incident warts. Among 501 HIV-seropositive and 43 HIV-seronegative women, warts regressed in 410 (82%) seropositive and 41 (95%) seronegative women (P=.02), most in the first year after diagnosis. In multivariable analysis, regression was negatively associated with HIV status and lower CD4 count as well as older age. Incident VIN of any grade occurred more frequently among HIV-seropositive than HIV-seronegative women: 0.42 (0.33-0.53) compared with 0.07 (0.02-0.18) per 100 person-years (P<.001). Positivity for VIN 2 was found in 58 women (55 with and three without HIV, P<.001). Two women with HIV developed stage IB squamous cell vulvar cancers. Although genital warts and VIN are more common among HIV-seropositive than HIV-seronegative women, wart regression is common even in women with HIV, and cancers are infrequent. ClinicalTrials.gov, www.clinicaltrials.gov, NCT00000797. II.


Diurnal variation around an optimum and near-critically high temperature does not alter the performance of an ectothermic aquatic grazer.
The growing threat of global climate change has led to a profusion of studies examining the effects of warming on biota. Despite the potential importance of natural variability such as diurnal temperature fluctuations, most experimental studies on warming are conducted under stable temperatures. Here, we investigated whether the responses of an aquatic invertebrate grazer (Lymnaea stagnalis) to an increased average temperature differ when the thermal regime is either constant or fluctuates diurnally. Using thermal response curves for several life-history and immune defense traits, we first identified the optimum and near-critically high temperatures that Lymnaea potentially experience during summer heat waves. We then exposed individuals that originated from three different populations to these two temperatures under constant or fluctuating thermal conditions. After 7 days, we assessed growth, reproduction, and two immune parameters (phenoloxidase-like activity and antibacterial activity of hemolymph) from each individual. Exposure to the near-critically high temperature led to increased growth rates and decreased antibacterial activity of hemolymph compared to the optimum temperature, whilst temperature fluctuations had no effect on these traits. The results indicate that the temperature level per se, rather than the variability in temperature was the main driver altering trait responses in our study species. Forecasting responses in temperature-related responses remains challenging, due to system-specific properties that can include intraspecific variation. However, our study indicates that experiments examining the effects of warming using constant temperatures can give similar predictions as studies with fluctuating thermal dynamics, and may thus be useful indicators of responses in nature.


An initial evaluation of gellan gum as a material for tissue engineering applications.
Alpha-modified minimum essential medium (alphaMEM) has been found to cross-link a 1% gellan gum solution, resulting in the formation of a self-supporting hydrogel in 1:1 and 5:1 ratios of polysaccharide: alphaMEM. Rheological data from temperature sweeps confirm that in addition to orders of magnitude differences in G' between 1% gellan and 1% gellan with alphaMEM, there is also a 20 degrees C increase in the temperature at which the onset of gelation takes place when alphaMEM is present. Frequency sweeps confirm the formation of a true gel; mechanical spectra for mixtures of gellan and alphaMEM clearly demonstrate G' to be independent of frequency. It is possible to immobilize cells within a three-dimensional (3D) gellan matrix that remain viable for up to 21 days in culture by adding a suspension of rat bone marrow cells (rBMC) in alphaMEM to 1% gellan solution. This extremely simple approach to cell immobilization within 3D constructs, made possible by the fact that gellan solutions cross-link in the presence of millimolar concentrations of cations, poses a very low risk to a cell population immobilized within a gellan matrix and thus indicates the potential of gellan for use as a tissue engineering scaffold.


Looking into tomorrow. Health workforce issues confronting physician assistants.
An important issue facing the physician assistant (PA) profession is how it can achieve a balance between supply of, and demand for, the services that PAs provide in the health workforce of the future. Recently, there has been debate and discussion about the implications of the recent expansion of the supply of PAs. In this article, we review and discuss (1) workforce data on physicians, PAs, and nurse practitioners and (2) projections of the number of these clinicians who will be trained in the future. We then analyze (1) data that describe the past 11 years of PA education and (2) data that address the experience of recent graduates of PA education programs who have sought employment. Some evidence suggests that, although demand has kept pace with expanding supply, a perception clearly exists that the PA job market may be tightening in some regions of the United States.


Expression analysis of the transmembrane mucin MUC20 in human corneal and conjunctival epithelia.
Cell surface mucins are a group of highly O-glycosylated transmembrane glycoproteins responsible for the protection of epithelial cells on mucosal surfaces. The aim of this study was to investigate the localization and regulation of mucin 20 (MUC20) at the ocular surface. Localization of MUC20 in human corneal and conjunctival epithelia was evaluated by immunofluorescence microscopy. Immortalized corneal (HCLE) and conjunctival (HCjE) cell lines were grown at different stages of differentiation and subjected to quantitative PCR and Western blot analyses. Cell surface proteins on apical cell membranes were biotinylated and isolated by neutravidin chromatography. The MUC20 was detected throughout the entire human ocular surface epithelia, predominantly in cell membranes within intermediate cell layers. In conjunctiva, MUC20 also was observed in the cytoplasm of apical cells within the stratified squamous epithelium, but not in goblet cells. Quantitative PCR and immunoblotting demonstrated expression of MUC20 in HCLE and HCjE cells. Induction of differentiation with serum-containing medium resulted in upregulation of MUC20 mRNA and protein. Biotin labeling of the surface of stratified cultures revealed low levels of MUC20 protein on apical glycocalyces. Further, MUC20 was not detected in the cell culture media or in human tears, suggesting that the extracellular domain of MUC20 is not released from the ocular surface as described previously for other cell surface mucins. Our results indicate that MUC20 is a novel transmembrane mucin expressed by the human corneal and conjunctival epithelia, and suggest that differential expression of MUC20 during differentiation has a role in maintaining ocular surface homeostasis.


Nanowell-trapped charged ligand-bearing nanoparticle surfaces: a novel method of enhancing flow-resistant cell adhesion.
Assuring cell adhesion to an underlying biomaterial surface is vital in implant device design and tissue engineering, particularly under circumstances where cells are subjected to potential detachment from overriding fluid flow. Cell-substrate adhesion is a highly regulated process involving the interplay of mechanical properties, surface topographic features, electrostatic charge, and biochemical mechanisms. At the nanoscale level, the physical properties of the underlying substrate are of particular importance in cell adhesion. Conventionally, natural, pro-adhesive, and often thrombogenic, protein biomaterials are frequently utilized to facilitate adhesion. In the present study, nanofabrication techniques are utilized to enhance the biological functionality of a synthetic polymer surface, polymethymethacrylate, with respect to cell adhesion. Specifically we examine the effect on cell adhesion of combining: 1. optimized surface texturing, 2. electrostatic charge and 3. cell adhesive ligands, uniquely assembled on the substrata surface, as an ensemble of nanoparticles trapped in nanowells. Our results reveal that the ensemble strategy leads to enhanced, more than simply additive, endothelial cell adhesion under both static and flow conditions. This strategy may be of particular utility for enhancing flow-resistant endothelialization of blood-contacting surfaces of cardiovascular devices subjected to flow-mediated shear.


Risk prediction of cardiovascular disease in type 2 diabetes: a risk equation from the Swedish National Diabetes Register.
Risk prediction models obtained in samples from the general population do not perform well in type 2 diabetic patients. Recently, 5-year risk estimates were proposed as being more accurate than 10-year risk estimates. This study presents a diabetes-specific equation for estimation of the absolute 5-year risk of first incident fatal/nonfatal cardiovascular disease (CVD) in type 2 diabetic patients with use of A1C and clinical characteristics. The study was based on 11,646 female and male patients, aged 18-70 years, from the Swedish National Diabetes Register with 1,482 first incident CVD events based on 58,342 person-years with mean follow-up of 5.64 years. This risk equation incorporates A1C, as in the UK Prospective Diabetes Study risk engine, and several clinical characteristics: onset age of diabetes, diabetes duration, sex, BMI, smoking, systolic blood pressure, and antihypertensive and lipid-reducing drugs. All predictors included were associated with the outcome (P < 0.0001, except for BMI P = 0.0016) with Cox regression analysis. Calibration was excellent when assessed by comparing observed and predicted risk. Discrimination was sufficient, with a receiver operator curve statistic of 0.70. Mean 5-year risk of CVD in all patients was 12.0 +/- 7.5%, whereas 54% of the patients had a 5-year risk >or=10%. This more simplified risk equation enables 5-year risk prediction of CVD based on easily available nonlaboratory predictors in clinical practice and A1C and was elaborated in a large observational study obtained from the normal patient population aged up to 70 years.


Use of green fluorescent protein for visualization of cell-specific gene expression and subcellular protein localization during sporulation in Bacillus subtilis.
We report the use of the green fluorescent protein (GFP) of Aequorea victoria to visualize cell-specific gene expression and protein subcellular localization during sporulation in Bacillus subtilis. Sporangia bearing the gene (gfp) for the green fluorescent protein fused to genes under the control of the sporulation transcription factor sigma F exhibited a forespore-specific pattern of fluorescence. Forespore-specific fluorescence could be detected with fusions to promoters that are utilized with low (csfB) and high (sspE-2G) efficiency by sigma F-containing RNA polymerase. Conversely, a mother cell-specific pattern of fluorescence was observed in sporangia bearing a transcriptional fusion of gfp to a spore coat protein gene (cotE) under the control of sigma E and an in-frame fusion to a regulatory gene (gerE) under the control of sigma K. An in-frame fusion of gfp to cotE demonstrated that GFP can also be used to visualize protein subcellular localization. In sporangia producing the CotE-GFP fusion protein, fluorescence was found to localize around the developing spore, and this localization was dependent upon SpoIVA, a morphogenetic protein known to determine proper localization of CotE.


Cystectomy for benign disease: readmission, morbidity, and complications.
We sought to elucidate outcomes and risks associated with cystectomy and urinary diversion for benign urological conditions compared to malignant conditions. We identified patients who underwent cystectomy and urinary diversion for benign and malignant diseases through the American College of Surgeons National Surgery Quality Improvement Program database for the period 2007-2015. Patients were selected for inclusion based upon their current procedure terminology and International Classification of Disease, Ninth revision codes. Primary outcome was 30 day morbidity including return to the operating room (OR); infectious, respiratory, and/or cardiovascular complications; readmission to the hospital; and mortality. Multivariable regression analyses were performed to identify associated factors. A total of 317 patients underwent cystectomy and urinary diversion for benign disease, and 5510 patients underwent radical cystectomy with urinary diversion for cancer. Rates of major morbidity (43.2% versus 38.6%), mortality (0.9% versus 1.9%), return to OR (5% versus 5.8%), readmission (19.7% versus 21.4%), postoperative sepsis (14.5% versus 12%), and wound complications (16.1% versus 14.2%) were similar among patients undergoing cystectomy for benign and malignant conditions. In the group with cystectomy for benign conditions, smoking (OR: 3.11) and longer operative duration (OR: 1.06) were significantly associated with increased overall morbidity. Wound complications were significantly higher in smokers (OR: 3.09) and with an ASA ≥ III (OR: 5.71) CONCLUSIONS: Patients undergoing cystectomy and urinary diversion for benign disease are at similar risk for 30 day morbidity and mortality as patients undergoing surgery for malignant conditions. Risk factors are identified that can potentially be targeted for morbidity reduction.


Within-Season Changes in Cytauxzoon felis Parasitemia in Bobcats.
Cytauxzoon felis is a pathogen responsible for cytauxzoonosis, a highly fatal disease in domestic cats. Although most studies of C. felis have focused on this parasite in domestic cats, bobcats are the reservoir host. In stark contrast, there is little information relative to the progression of C. felis infections in bobcats. We studied bobcats in southern Illinois during 2014-2017 to evaluate which environmental factors (i.e., ambient temperature; number of daylight hours; trapping year, month, and day) influenced C. felis parasitemia levels. Mean ambient temperature at 1 wk and 2 wk prior to sampling was associated with increased parasitemia levels. Vector activity intensifies with higher temperatures, suggesting that increased parasitemia levels are an adaptation to facilitate transmission.


A clinician's guide to patient selection for watchful waiting management of inguinal hernia.
The goal of this study was to assist surgeons in managing patients with minimally symptomatic inguinal hernia by identifying characteristics that predict crossover to surgery or worsening of hernia symptoms. Randomized trials have suggested that watchful waiting management of minimally symptomatic inguinal hernia is an acceptable alternative to surgical repair. However, these trials found that roughly a quarter of patients would elect for repair in the first 2 years, suggesting that not all patients are good candidates for watchful waiting. The 336 patients randomized to watchful waiting in the American College of Surgeons Watchful Waiting Hernia Trial constituted the study population. Preoperative patient characteristics were used to predict 2 outcomes, either crossover to surgery or the development of hernia pain limiting activities and/or crossover to surgery. Patients in our study were part of a previously registered randomized trial: NCT00263250. At 2 years, 72 patients crossed over to surgery, with pain with strenuous activities [odds ratio (OR), 1.3 per 10-mm visual analog scale pain scale], chronic constipation (OR, 4.9), prostatism (OR, 2.9), being married (OR, 2.3), and good health [OR, 3.0 American Society of Anesthesiologists Class (ASA) 1 vs 2], predicting crossover. An additional 28 patients developed pain, limiting their activities, with pain during strenuous activities (OR, 1.3 per 10-mm visual analog scale) and chronic constipation (OR, 4.5), predicting the combined outcome of pain limiting activities and/or crossover to surgery. Higher levels of activity reduced the risk (OR, 0.95) of this combined outcome. Readily identifiable patient characteristics can predict those patients with minimally symptomatic inguinal hernia who are likely to "fail" watchful waiting hernia management. Consideration of these factors will allow surgeons to optimally tailor hernia management.


Gray Matter Structural Alterations in Chronic and Episodic Migraine: A Morphometric Magnetic Resonance Imaging Study.
This study evaluates different parameters describing the gray matter structure to analyze differences between healthy controls, patients with episodic migraine, and patients with chronic migraine. Cohort study. Spanish community. Fifty-two healthy controls, 57 episodic migraine patients, and 57 chronic migraine patients were included in the study and underwent T1-weighted magnetic resonance imaging acquisition. Eighty-four cortical and subcortical gray matter regions were extracted, and gray matter volume, cortical curvature, thickness, and surface area values were computed (where applicable). Correlation analysis between clinical features and structural parameters was performed. Statistically significant differences were found between all three groups, generally consisting of increases in cortical curvature and decreases in gray matter volume, cortical thickness, and surface area in migraineurs with respect to healthy controls. Furthermore, differences were also found between chronic and episodic migraine. Significant correlations were found between duration of migraine history and several structural parameters. Migraine is associated with structural alterations in widespread gray matter regions of the brain. Moreover, the results suggest that the pattern of differences between healthy controls and episodic migraine patients is qualitatively different from that occurring between episodic and chronic migraine patients.


Status of hepatocellular carcinoma in Gulf region.
Hepatocellular carcinoma (HCC) has a unique geographic distribution that is likely to be determined by specific etiologic factors. There is a distinctive difference in sex and age related occurrence of disease. In the Gulf region, there are contradicting data on the prevalence and death rates due to HCC. In this review we highlight some aspects of HCC specific to the Gulf region. A retrospective analysis of 150 patient's data is presented, including demographic, epidemiological, aetiological disease status assessment with child Pugh criteria, modes of treatment and treatment related outcome. Hepatitis C virus (HCV) infection was the most common (45%) documented etiology, similar to Western European countries, followed by hepatitis B virus (HBV) infection in 27% of cases, alcoholic liver disease only in six patients (4%). Child-Pugh assessment was A in 33%, B in 37% and C in 30% of observed patients. Surgery (liver resection or transplantation) was performed in 12% and local ablation in 5% of cases. The others were treated by chemo-embolization in 17% and by systemic therapy with sorafenib in 13% of patients. Nearly half of the patients (53%) were in advanced stages and received palliative treatment. To improve the outcome of treatment in HCC patients in the Gulf region, an effective and strategic screening program must be implemented for early diagnosis and treatment to improve the outcome of this mostly fatal disease.


A novel method utilising markers of altered erythropoiesis for the detection of recombinant human erythropoietin abuse in athletes.
The use of recombinant human erythropoietin (r-HuEPO) to enhance athletic performance is prohibited. Existing tests cannot readily differentiate between exogenous and endogenous EPO. Therefore the aim of our study was to investigate possible indirect detection of r-HuEPO use via blood markers of altered erythropoiesis. Twenty-seven recreational athletes were assigned to three groups prior to a 25 day drug administration phase, with the following protocols: EPO+IM group (n = 10), 50 Ukg(-1) r-HuEPO at a frequency of 3wk(-1), 100 mg intramuscular (IM) iron 1wk(-1) and a sham iron tablet daily; EPO+OR group (n = 8), 50 U.kg(-1) r-HuEPO 3wk(-1), sham iron injection 1wk(-1) and 105 mg of oral elemental iron daily; placebo group (n = 9), sham r-HuEPO injections 3wk(-1), sham iron injections 1wk(-1) and sham iron tablets daily. Each group was monitored during and for 4 weeks after drug administration. Models incorporating combinations of the variables reticulocyte hematocrit (RetHct), serum EPO, soluble transferrin receptor, hematocrit (Hct) and % macrocytes were analyzed by logistic regression. One model (ON-model) repeatedly identified 94-100% of r-HuEPO group members during the final 2 wk of the r-HuEPO administration phase. One false positive was recorded from a possible 189. Another model (OFF-model) incorporating RetHct, EPO and Hct was applied during the wash-out phase and, during the period of 12-21 days after the last r-HuEPO injection, it repeatedly identified 67-72% of recent users with no false positives. Multiple indirect hematologic and biochemical markers used simultaneously are potentially effective for identifying current or recent users of r-HuEPO.


Expression of growth-differentiating factor 9 and its type 1 receptor in human ovaries.
The expression of growth-differentiating factor 9 (GDF9) has not been studied in ovaries from girls and human fetuses nor has its receptor transforming growth factor-beta1 receptor (TGFbetaR1) been investigated in ovaries of girls/women. The aim of this study was to fill these gaps. Ovarian samples were obtained from 16 human fetuses at 21-35 gestational weeks and from 34 girls/women aged 5-39years. These specimens were prepared for immunohistochemical staining of the GDF9 and TGFbetaR1 proteins. Reverse transcription polymerase chain reaction was used to detect GDF9 mRNA transcripts and in-situ hybridization to localize TGFbetaR1 mRNA transcripts. Positive staining for the GDF9 protein was identified in oocytes and granulosa cells in all samples tested. GDF9 mRNA transcripts were present in all samples. Protein expression of TGFbetaR1 was identified in granulosa cells in all samples. Oocyte staining was identified in samples from girls/women but in only one fetal sample. TGFbetaR1 mRNA transcripts were identified in granulosa cells and oocytes in 50% of the samples from fetuses aged over 22 gestational weeks and in samples from girls/women. The detection of GDF9 and TGFbetaR1 at both at the protein and mRNA levels suggests that GDF9 may have functions in human preantral follicles.


Srsf2P95H initiates myeloid bias and myelodysplastic/myeloproliferative syndrome from hemopoietic stem cells.
Mutations in SRSF2 occur in myelodysplastic syndromes (MDS) and MDS/myeloproliferative neoplasms (MPN). SRSF2 mutations cluster at proline 95, with the most frequent mutation being a histidine (P95H) substitution. They undergo positive selection, arise early in the course of disease, and have been identified in age-related clonal hemopoiesis. It is not clear how mutation of SRSF2 modifies hemopoiesis or contributes to the development of myeloid bias or MDS/MPN. Two prior mouse models of Srsf2P95H mutation have been reported; however, these models do not recapitulate many of the clinical features of SRSF2-mutant disease and relied on bone marrow (BM) transplantation stress to elicit the reported phenotypes. We describe a new conditional murine Srsf2P95H mutation model, where the P95H mutation is expressed physiologically and heterozygously from its endogenous locus after Cre activation. Using multiple Cre lines, we demonstrate that during native hemopoiesis (ie, no BM transplantation), the Srsf2P95H mutation needs to occur within the hemopoietic stem-cell-containing populations to promote myelomonocytic bias and expansion with corresponding transcriptional and RNA splicing changes. With age, nontransplanted Srsf2P95H animals developed a progressive, transplantable disease characterized by myeloid bias, morphological dysplasia, and monocytosis, hallmarks of MDS/MPN in humans. Analysis of cooccurring mutations within the BM demonstrated the acquisition of additional mutations that are recurrent in humans with SRSF2 mutations. The tractable Srsf2P95H/+ knock-in model we have generated is highly relevant to human disease and will serve to elucidate the effect of SRSF2 mutations on initiation and maintenance of MDS/MPN.


Fluorine-18 labelled building blocks for PET tracer synthesis.
Positron emission tomography (PET) is an important driver for present day healthcare. Fluorine-18 is the most widely used radioisotope for PET imaging and a thorough overview of the available radiochemistry methodology is a prerequisite for selection of a synthetic approach for new fluorine-18 labelled PET tracers. These PET tracers can be synthesised either by late-stage radiofluorination, introducing fluorine-18 in the last step of the synthesis, or by a building block approach (also called modular build-up approach), introducing fluorine-18 in a fast and efficient manner in a building block, which is reacted further in one or multiple reaction steps to form the PET tracer. This review presents a comprehensive overview of the synthesis and application of fluorine-18 labelled building blocks since 2010.


NPAS2 as a transcriptional regulator of non-rapid eye movement sleep: genotype and sex interactions.
Because the transcription factor neuronal Per-Arnt-Sim-type signal-sensor protein-domain protein 2 (NPAS2) acts both as a sensor and an effector of intracellular energy balance, and because sleep is thought to correct an energy imbalance incurred during waking, we examined NPAS2's role in sleep homeostasis using npas2 knockout (npas2-/-) mice. We found that, under conditions of increased sleep need, i.e., at the end of the active period or after sleep deprivation (SD), NPAS2 allows for sleep to occur at times when mice are normally awake. Lack of npas2 affected electroencephalogram activity of thalamocortical origin; during non-rapid eye movement sleep (NREMS), activity in the spindle range (10-15 Hz) was reduced, and within the delta range (1-4 Hz), activity shifted toward faster frequencies. In addition, the increase in the cortical expression of the NPAS2 target gene period2 (per2) after SD was attenuated in npas2-/- mice. This implies that NPAS2 importantly contributes to the previously documented wake-dependent increase in cortical per2 expression. The data also revealed numerous sex differences in sleep; in females, sleep need accumulated at a slower rate, and REMS loss was not recovered after SD. In contrast, the rebound in NREMS time after SD was compromised only in npas2-/- males. We conclude that NPAS2 plays a role in sleep homeostasis, most likely at the level of the thalamus and cortex, where NPAS2 is abundantly expressed.


Speech-on-speech masking with variable access to the linguistic content of the masker speech for native and nonnative english speakers.
Masking release for an English sentence-recognition task in the presence of foreign-accented English speech compared with native-accented English speech was reported in Calandruccio et al (2010a). The masking release appeared to increase as the masker intelligibility decreased. However, it could not be ruled out that spectral differences between the speech maskers were influencing the significant differences observed. The purpose of the current experiment was to minimize spectral differences between speech maskers to determine how various amounts of linguistic information within competing speech Affiliationect masking release. A mixed-model design with within-subject (four two-talker speech maskers) and between-subject (listener group) factors was conducted. Speech maskers included native-accented English speech and high-intelligibility, moderate-intelligibility, and low-intelligibility Mandarin-accented English. Normalizing the long-term average speech spectra of the maskers to each other minimized spectral differences between the masker conditions. Three listener groups were tested, including monolingual English speakers with normal hearing, nonnative English speakers with normal hearing, and monolingual English speakers with hearing loss. The nonnative English speakers were from various native language backgrounds, not including Mandarin (or any other Chinese dialect). Listeners with hearing loss had symmetric mild sloping to moderate sensorineural hearing loss. Listeners were asked to repeat back sentences that were presented in the presence of four different two-talker speech maskers. Responses were scored based on the key words within the sentences (100 key words per masker condition). A mixed-model regression analysis was used to analyze the difference in performance scores between the masker conditions and listener groups. Monolingual English speakers with normal hearing benefited when the competing speech signal was foreign accented compared with native accented, allowing for improved speech recognition. Various levels of intelligibility across the foreign-accented speech maskers did not influence results. Neither the nonnative English-speaking listeners with normal hearing nor the monolingual English speakers with hearing loss benefited from masking release when the masker was changed from native-accented to foreign-accented English. Slight modifications between the target and the masker speech allowed monolingual English speakers with normal hearing to improve their recognition of native-accented English, even when the competing speech was highly intelligible. Further research is needed to determine which modifications within the competing speech signal caused the Mandarin-accented English to be less effective with respect to masking. Determining the influences within the competing speech that make it less effective as a masker or determining why monolingual normal-hearing listeners can take advantage of these differences could help improve speech recognition for those with hearing loss in the future.


Structural analysis of strand misalignment during DNA synthesis by a human DNA polymerase.
Insertions and deletions in coding sequences can alter the reading frame of genes and have profound biological consequences. In 1966, Streisinger proposed that these mutations result from strand slippage, which in repetitive sequences generates misaligned intermediates stabilized by correct base pairing that support polymerization. We report here crystal structures of human DNA polymerase lambda, which frequently generates deletion mutations, bound to such intermediates. Each contains an extrahelical template nucleotide upstream of the active site. Surprisingly, the extra nucleotide, even when combined with an adjacent mismatch, does not perturb polymerase active site geometry, which is indistinguishable from that for correctly aligned strands. These structures reveal how pol lambda can polymerize on substrates with minimal homology during repair of double-strand breaks and represent strand-slippage intermediates consistent with Streisinger's classical hypothesis. They are thus relevant to the origin of single-base deletions, a class of mutations that can confer strong biological phenotypes.


Water and ion transport by the urinary bladder of the teleost Pseudopleuronectes americanus.
Water and ion transport by the isolated teleost urinary bladder were studied. The transepithelial electrical PD across sac-type bladder preparations was unstable, i.e., initially mucosa positive but becoming more negative with time. Perfused bladders maintained a low mucosa positive PD which was stable. Both Na and Cl appeared to be actively transported from mucosal side (M) to serosal side (S). Voltage clamping the bladder at 0, -50, or +50 mV had almost no effect on active or passive Na or Cl flux in either direction. Na and Cl transport seemed electrically neutral. Fluid absorption (M to S) was directly correlated with absorption of osmotically active solutes. These solutes were almost all Na and Cl. The bladder acidified and secreted K+ into the mucosal fluid. Divalent ions were concentrated in the mucosal fluid as a result of fluid absorption. Although furosemide and ethacrynic acid inhibited ion and water transport by the bladder, ouabain was effective at a much lower concentration. Ouabain (10(-4) M) inhibited active Na transport when applied only to the mucosal or only to the serosal surface. Ouabain abolished the PD only from the serosal surface.


Hepatitis C virus infection and diabetes mellitus in end-stage renal disease: evidence of a negative association.
Epidemiological studies have implicated hepatitis C virus (HCV) infection in the pathogenesis of diabetes mellitus (DM) both in the population as a whole and after solid organ transplantation. Whether this association exists in patients with end-stage renal disease (ESRD) undergoing dialysis is unclear. The aim of this study is to investigate the relationship between HCV and DM in a large group (n=742) of patients with ESRD from Europe and North America. The presence of diabetes was ascertained by using American Diabetes Association guidelines based on fasting glucose measurement and medication history. Presence of HCV infection was assessed by serum testing for anti-HCV antibodies. The prevalence of anti-HCV antibody positive patients was 15% (112/742); the frequency of DM was higher among anti-HCV positive than -HCV negative patients but the difference did not approach statistical significance, 32% (36/112) vs 29.5% (186/630). The frequency of patients with diabetic nephropathy was not higher in anti-HCV positive than -negative patients; 21.4% (24/112) vs 23.3% (147/630), NS. Logistic regression model showed an independent and significant link between anti-HCV seropositive status and raised GPT (P=0.032), male gender (P = 0.0462), positive history of prior renal transplant (P=0.0006), and longer time on dialysis (P=0.00001). In summary, no link between anti-HCV antibody and DM occurred in this ESRD population; there was no association between rate of anti-HCV antibody and diabetic nephropathy.


A systematic review of the effectiveness and cost-effectiveness of neuroimaging assessments used to visualise the seizure focus in people with refractory epilepsy being considered for surgery.
To review the effectiveness and/or accuracy, cost-effectiveness, and predictive value of neuroimaging of the cerebral cortex to visualise the seizure focus in people with refractory epilepsy being considered for surgery. Electronic databases, Internet searches, hand searching and consultation with experts. A systematic review was undertaken according to published guidelines. Results of diagnostic accuracy studies were analysed according to the imaging test evaluated. For each study the proportion of patients who were correctly localised, not localised, partially localised or incorrectly localised by the index test was calculated. Due to the heterogeneity present between studies, statistical pooling was not performed. Instead, a narrative synthesis of results is presented. For studies using multivariate analysis to look at the association of neuroimaging findings and outcome following surgery, all factors considered in the analyses were presented. Studies were grouped according to the neuroimaging technique investigated and the findings discussed with reference to possible sources of heterogeneity between studies. No randomised controlled trials (RCTs) were identified, with the majority of studies evaluating the diagnostic accuracy of various imaging techniques in the localisation of epileptic seizure foci. There was significant heterogeneity (p<0.05) between studies for at least one of the localisation categories (correctly localised, not localised, partially localised and incorrectly localised) for all imaging techniques. Possible explanations for this heterogeneity include differing study designs, index test characteristics, reference standards and population characteristics. Test performance was more promising in studies restricted to patients with temporal lobe epilepsy. Ictal single photon emission computed tomography (SPECT) generally had more correctly localising (70--100%) and fewer non-localising (0--7%) scans than other techniques evaluated in patients with temporal lobe epilepsy. Results for computed tomography and interictal SPECT suggest that these tests are relatively poor at localising the seizure focus. Volumeric magnetic resonance imaging (MRI) and position emission tomography (PET) appear promising, and subtraction ictal single photon emission computed tomography co-registered to magnetic resonance imaging (SISCOM) and magnetic resonance spectrosopy (MRS) less promising than ictal SPECT, but these technologies have been assessed in fewer studies. T2 relaxometry was reported in only one small study with inconclusive results. Seventeen studies (33 evaluations) provided sufficient data on the association of a localised scan with outcome following surgery to calculate a relative risk. The majority of evaluations (24/33) suggested that patients with a correctly or partially localised scan had a better outcome following surgery than those with an incorrectly localised or non-localised scan. However, this association was statistically significant in only three studies, two evaluating routine MRI [(relative risk (RR) 2.74, 95% confidence interval (CI): 1.32 to 5.67; RR 1.28, 95% CI: 1.00 to 1.63] and the other SISCOM (RR 2.12, 95% CI: 1.01, 4.44). Nine studies used multivariate analysis to investigate the association of MRI (7 studies), MRS and volumetric MRI (1 study), PET (3 studies), SPECT (1 study) and SISCOM (3 studies) with the outcome following surgery. There was a trend for localisation of abnormalities to be associated with a beneficial outcome. Due to the limitations of the included studies, the results of this review do little to inform clinical practice, with insufficient evidence regarding effectiveness and cost-effectiveness of imaging techniques in the work-up for epilepsy surgery. Given the inadequacy of existing data, there is a pressing need for studies investigating the utility of imaging techniques in the work up for epilepsy surgery. The most reliable method to achieve this is the RCT, which could examine the single tests or combinations of tests on patient outcome. The authors suggest that it is important that clinicians, patient groups, policy makers and healthcare/research funders meet and debate the most appropriate way to investigate these technologies.


Thermal characterization and analysis of microliter liquid volumes using the three-omega method.
Thermal phenomena in many biological systems offer an alternative detection opportunity for quantifying relevant sample properties. While there is substantial prior work on thermal characterization methods for fluids, the push in the biology and biomedical research communities towards analysis of reduced sample volumes drives a need to extend and scale these techniques to these volumes of interest, which can be below 100 pl. This work applies the 3ω technique to measure the temperature-dependent thermal conductivity and heat capacity of de-ionized water, silicone oil, and salt buffer solution droplets from 24 to 80 °C. Heater geometries range in length from 200 to 700 μm and in width from 2 to 5 μm to accommodate the size restrictions imposed by small volume droplets. We use these devices to measure droplet volumes of 2 μl and demonstrate the potential to extend this technique down to pl droplet volumes based on an analysis of the thermally probed volume. Sensitivity and uncertainty analyses provide guidance for relevant design variables for characterizing properties of interest by investigating the tradeoffs between measurement frequency regime, device geometry, and substrate material. Experimental results show that we can extract thermal conductivity and heat capacity with these sample volumes to within less than 1% of thermal properties reported in the literature.


The default modes of reading: modulation of posterior cingulate and medial prefrontal cortex connectivity associated with comprehension and task focus while reading.
Reading is a fundamental human capacity and yet it can easily be derailed by the simple act of mind-wandering. A large-scale brain network, referred to as the default mode network (DMN), has been shown to be involved in both mind-wandering and reading, raising the question as to how the same neural system could be implicated in processes with both costs and benefits to narrative comprehension. Resting-state functional magnetic resonance imaging (rs-fMRI) was used to explore whether the intrinsic functional connectivity of the two key midline hubs of the DMN-the posterior cingulate cortex (PCC) and anterior medial prefrontal cortex (aMPFC)-was predictive of individual differences in reading comprehension and task focus recorded outside of the scanner. Worse comprehension was associated with greater functional connectivity between the PCC and a region of the ventral striatum. Better comprehension was associated with greater functional connectivity with a region of the right insula. By contrast reports of increasing task focus were associated with functional connectivity from the aMPFC to clusters in the PCC, the left parietal and temporal cortex, and the cerebellum. Our results suggest that the DMN has both costs (such as poor comprehension) and benefits to reading (such as an on-task focus) because its midline core can couple its activity with other regions to form distinct functional communities that allow seemingly opposing mental states to occur. This flexible coupling allows the DMN to participate in cognitive states that complement the act of reading as well as others that do not.


Spectrophotometric analysis of the expression of secreted aspartyl proteinases from Candida in leukoplakia and oral squamous cell carcinoma.
Candida species are a normal commensal of the oral cavity in healthy individuals, but can become an opportunistic pathogen when the oral ecosystem is unbalanced. Several virulence attributes have been identified in candidal infection, among which are the hydrolases, including the secreted aspartyl proteinases (Saps). This study evaluated and compared the in vitro level of Saps from Candida albicans in nonsmokers, smokers, and patients with leukoplakia and oral squamous cell carcinoma (OSCC). Candida cell count (CCC) at 48 h was also assessed. The Sap level was measured by spectrophotometry in 38 clinical isolates of C. albicans obtained from the oral cavity of the four different groups. Culturing was done in yeast carbon base-bovine serum albumin. Speciation of Candida was performed by using a Candida identification kit, and CCC was measured by hemocytometer. Sap levels and CCC were higher in individuals with leukoplakia and OSCC than in nonsmokers or smokers (P = 0.001); however, there was no significant difference in Sap levels or CCC between smokers and nonsmokers (P = 0.529). Further, an intragroup correlation between CCC and Sap level was also observed. The higher level of Saps from C. albicans in individuals with leukoplakia and OSCC suggests that this pathogen plays a role in disease development and could aid in identifying the pathogenic commensal.


Curcumin-containing diet inhibits diethylnitrosamine-induced murine hepatocarcinogenesis.
Curcumin has been widely used as a spice and coloring agent in foods. Recently, curcumin was found to possess chemopreventive effects against skin cancer, forestomach cancer, colon cancer and oral cancer in mice. Clinical trials of curcumin for prevention of human cancers are currently ongoing. In this study, we examine the chemopreventive effect of curcumin on murine hepatocarcinogenesis. C3H/HeN mice were injected i.p. with N-diethylnitrosamine (DEN) at the age of 5 weeks. The curcumin group started eating 0.2% curcumin-containing diet 4 days before DEN injection until death. The mice were then serially killed at the scheduled times to examine the development of hepatocellular carcinoma (HCC) and changes in intermediate biological markers. At the age of 42 weeks, the curcumin group, as compared with the control group (DEN alone), had an 81% reduction in multiplicity (0.5 versus 2.57) and a 62% reduction in incidence (38 versus 100%) of development of HCC. A series of intermediate biological markers were examined by western blot. While hepatic tissues obtained from the DEN-treated mice showed a remarkable increase in the levels of p21(ras), PCNA and CDC2 proteins, eating a curcumin-containing diet reversed the levels to normal values. These results indicate that curcumin effectively inhibits DEN-induced hepatocarcinogenesis in the mouse. The underlying mechanisms of the phenomenon and the feasibility of using curcumin in the chemoprevention of human HCC should be further explored.


Phenotypic convergence in genetically distinct lineages of a Rhinolophus species complex (Mammalia, Chiroptera).
Phenotypes of distantly related species may converge through adaptation to similar habitats and/or because they share biological constraints that limit the phenotypic variants produced. A common theme in bats is the sympatric occurrence of cryptic species that are convergent in morphology but divergent in echolocation frequency, suggesting that echolocation may facilitate niche partitioning, reducing competition. If so, allopatric populations freed from competition, could converge in both morphology and echolocation provided they occupy similar niches or share biological constraints. We investigated the evolutionary history of a widely distributed African horseshoe bat, Rhinolophus darlingi, in the context of phenotypic convergence. We used phylogenetic inference to identify and date lineage divergence together with phenotypic comparisons and ecological niche modelling to identify morphological and geographical correlates of those lineages. Our results indicate that R. darlingi is paraphyletic, the eastern and western parts of its distribution forming two distinct non-sister lineages that diverged ~9.7 Mya. We retain R. darlingi for the eastern lineage and argue that the western lineage, currently the sub-species R. d. damarensis, should be elevated to full species status. R. damarensis comprises two lineages that diverged ~5 Mya. Our findings concur with patterns of divergence of other co-distributed taxa which are associated with increased regional aridification between 7-5 Mya suggesting possible vicariant evolution. The morphology and echolocation calls of R. darlingi and R. damarensis are convergent despite occupying different biomes. This suggests that adaptation to similar habitats is not responsible for the convergence. Furthermore, R. darlingi forms part of a clade comprising species that are bigger and echolocate at lower frequencies than R. darlingi, suggesting that biological constraints are unlikely to have influenced the convergence. Instead, the striking similarity in morphology and sensory biology are probably the result of neutral evolutionary processes, resulting in the independent evolution of similar phenotypes.


Inhibition of rat brain monoamine oxidase enzymes by fluoxetine and norfluoxetine.
Fluoxetine and its primary metabolite, norfluoxetine, are inhibitors of neuronal uptake of 5-hydroxytryptamine. While fluoxetine has also been reported to inhibit monoamine oxidase (MAO) in vitro at concentrations much lower than those measured in brain following chronic fluoxetine treatment, neurochemical profiles are not consistent with substantial MAO inhibition in vivo. In an attempt to explain this inconsistency, we have examined the interactions of fluoxetine and norfluoxetine with rat brain MAO-A and -B by a radiochemical assay method. Fluoxetine and norfluoxetine were competitive inhibitors of MAO-A in vitro, with Ki values of 76.3 microM and 90.5 microM, respectively. Both compounds were non-competitive or uncompetitive inhibitors of MAO-B in vitro. Inhibition of MAO-B was time-dependent and was very slowly reversible by dialysis. IC50 values versus metabolism of 50 microM beta-phenylethylamine were 17.8 microM (fluoxetine) and 18.5 microM (norfluoxetine). Analysis of the time-dependence of MAO-B inhibition by fluoxetine revealed that an initial competitive interaction between the enzyme and the inhibitor (Ki 245 microM) was followed by tight-binding enzyme inactivation (K(inact) 0.071 min-1). Following administration of fluoxetine (20 mg kg-1 day-1) for 7 days, the cortical concentration of fluoxetine + norfluoxetine was estimated by gas-liquid chromatography to be 700 microM. Such drug treatment reduced MAO-A activity by 23% in 1:8 (w/v) cortical homogenates, but not in 1:80 homogenates. Inhibition of MAO-B in 1:8 homogenates was modest (12%) and was not significantly reduced by homogenate dilution. The concentration of 5-hydroxyindole-3-acetic acid, measured by high pressure liquid chromatography, was reduced by 47% in cortices from drug-treated rats, while concentrations of 5-hydroxytryptamine, noradrenaline, dopamine, 3,4-dihydroxyphenylacetic acid and homovanillic acid were unchanged. These results suggest that, following chronic drug administration leading to relatively high tissue concentrations of fluoxetine and norfluoxetine, inhibition of either form of MAO would be restricted by competition for the enzyme with intraneuronal amine substrates.


Prevalence, trends and associated socio-economic factors of obesity in South Asia.
Worldwide obesity levels have increased unprecedentedly over the past couple of decades. Although the prevalence, trends and associated socio-economic factors of the condition have been extensively reported in Western populations, less is known regarding South Asian populations. A review of articles using Medline with combinations of the MeSH terms: 'Obesity', 'Overweight' and 'Abdominal Obesity' limiting to epidemiology and South Asian countries. Despite methodological heterogeneity and variation according to country, area of residence and gender , the most recent nationally representative and large regional data demonstrates that without any doubt there is a epidemic of obesity, overweight and abdominal obesity in South Asian countries. Prevalence estimates of overweight and obesity (based on Asian cut-offs: overweight ≥ 23 kg/m², obesity ≥ 25 kg/m²) ranged from 3.5% in rural Bangladesh to over 65% in the Maldives. Abdominal obesity was more prevalent than general obesity in both sexes in this ethnic group. Countries with the lowest prevalence had the highest upward trend of obesity. Socio-economic factors associated with greater obesity in the region included female gender, middle age, urban residence, higher educational and economic status. South Asia is significantly affected by the obesity epidemic. Collaborative public health interventions to reverse these trends need to be mindful of many socio-economic constraints in order to provide long-term solutions.


A molecular dynamics study of slow base flipping in DNA using conformational flooding.
Individual DNA bases are known to be able to flip out of the helical stack, providing enzymes with access to the genetic information otherwise hidden inside the helix. Consequently, base flipping is a necessary first step to many more complex biological processes such as DNA transcription or replication. Much remains unknown about this elementary step, despite a wealth of experimental and theoretical studies. From the theoretical point of view, the involved timescale of milliseconds or longer requires the use of enhanced sampling techniques. In contrast to previous theoretical studies employing umbrella sampling along a predefined flipping coordinate, this study attempts to induce flipping without prior knowledge of the pathway, using information from a molecular dynamics simulation of a B-DNA fragment and the conformational flooding method. The relevance to base flipping of the principal components of the simulation is assayed, and a combination of modes optimally related to the flipping of the base through either helical groove is derived for each of the two bases of the central guanine-cytosine basepair. By applying an artificial flooding potential along these collective coordinates, the flipping mechanism is accelerated to within the scope of molecular dynamics simulations. The associated free energy surface is found to feature local minima corresponding to partially flipped states, particularly relevant to flipping in isolated DNA; further transitions from these minima to the fully flipped conformation are accelerated by additional flooding potentials. The associated free energy profiles feature similar barrier heights for both bases and pathways; the flipped state beyond is a broad and rugged attraction basin, only a few kcal/mol higher in energy than the closed conformation. This result diverges from previous works but echoes some aspects of recent experimental findings, justifying the need for novel approaches to this difficult problem: this contribution represents a first step in this direction. Important structural factors involved in flipping, both local (sugar-phosphate backbone dihedral angles) and global (helical axis bend), are also identified.


Effect of acute administration of gliclazide on the glucose sensitivity of pancreatic B-cells in healthy subjects.
1. Sulphonylurea derivatives are commonly used in the treatment of non-insulin-dependent diabetes mellitus. It is, however, unclear whether the hypoglycaemic effect of sulphonylurea derivatives is additive to the effect of glucose, or whether sulphonylurea derivatives act by increasing B-cell glucose sensitivity. 2. We assessed the effect of gliclazide on glucose-stimulated insulin secretion in eight healthy volunteers. Sixty minute hyperglycaemic glucose clamps (blood glucose levels of 8, 11 and 32 mmol/l) were performed, with and without prior administration of gliclazide (80 mg) 90 min before the glucose clamp. 3. Dose-response characteristics were assessed with a modified Michaelis-Menten equation. The Vmax. (maximal B-cell responsiveness) was not significantly changed (1.5 +/- 0.1 versus 1.3 +/- 0.2 and 5.0 +/- 0.5 versus 4.8 +/- 0.5 mmol/l for the first- and second-phase insulin secretion, respectively), whereas the ED50 (half-maximally stimulating blood glucose concentration) was significantly decreased by gliclazide for first-phase insulin secretion (7.6 +/- 0.3 versus 9.1 +/- 0.6 mmol/l) but not for second-phase insulin secretion (12.0 +/- 0.5 versus 12.3 +/- 0.5 mmol/l). 4. We conclude that gliclazide indeed leads to a shift to the left of the dose-response curve of first-phase insulin release in vivo without a change in Vmax, which indicates an apparent enhancement of B-cell glucose sensitivity.


Developing a Health-Related Quality-of-Life Measure for People With Intellectual Disability.
Using principles of community-based participatory research we developed a new theory-based measure of health-related quality of life (HRQOL) for individuals with intellectual disability (ID). We recruited adults with ID (n = 129) to take part in interviews and review successive versions of HRQOL items. Critical input about content and understandability shaped the items, as did input from four focus groups of parents/caregivers (n = 16) and representative stakeholders from community-based agencies (n = 7). The resulting HRQOL measure, called the HRQOL-IDD, contains 42 items. The response format depicts a gradient of fluid-filled cups ("none" to "full") to represent frequency of experience of each item on a 5-point scale.


PLK1 phosphorylates PAX3-FOXO1, the inhibition of which triggers regression of alveolar Rhabdomyosarcoma.
Pediatric tumors harbor very low numbers of somatic mutations and therefore offer few targets to improve therapeutic management with targeted drugs. In particular, outcomes remain dismal for patients with metastatic alveolar rhabdomyosarcoma (aRMS), where the chimeric transcription factor PAX3/7-FOXO1 has been implicated but problematic to target. In this report, we addressed this challenge by developing a two-armed screen for druggable upstream regulatory kinases in the PAX3/7-FOXO1 pathway. Screening libraries of kinome siRNA and small molecules, we defined PLK1 as an upstream-acting regulator. Mechanistically, PLK1 interacted with and phosphorylated PAX3-FOXO1 at the novel site S503, leading to protein stabilization. Notably, PLK1 inhibition led to elevated ubiquitination and rapid proteasomal degradation of the PAX3-FOXO1 chimeric oncoprotein. On this basis, we embarked on a preclinical validation of PLK1 as a target in a xenograft mouse model of aRMS, where the PLK1 inhibitor BI 2536 reduced PAX3-FOXO1-mediated gene expression and elicited tumor regression. Clinically, analysis of human aRMS tumor biopsies documented high PLK1 expression to offer prognostic significance for both event-free survival and overall survival. Taken together, these preclinical studies validate the PLK1-PAX3-FOXO1 axis as a rational target to treat aRMS.


How patch configuration affects the impact of disturbances on metapopulation persistence.
Disturbances affect metapopulations directly through reductions in population size and indirectly through habitat modification. We consider how metapopulation persistence is affected by different disturbance regimes and the way in which disturbances spread, when metapopulations are compact or elongated, using a stochastic spatially explicit model which includes metapopulation and habitat dynamics. We discover that the risk of population extinction is larger for spatially aggregated disturbances than for spatially random disturbances. By changing the spatial configuration of the patches in the system--leading to different proportions of edge and interior patches--we demonstrate that the probability of metapopulation extinction is smaller when the metapopulation is more compact. Both of these results become more pronounced when colonization connectivity decreases. Our results have important management implication as edge patches, which are invariably considered to be less important, may play an important role as disturbance refugia.


Community engagement in the development and implementation of a rural road safety campaign: Steps and lessons learned.
Rural crashes result in fatality rates twice as high as urban, after accounting for vehicle miles traveled, and those involving farm vehicles tend to be the most severe. Farm vehicle crash interventions have focused on the farm equipment (e.g., lighting and marking) or the farm vehicle operator (e.g., training), despite crashes being most frequently caused by other vehicle driver actions. Community-based campaigns focused on rural drivers have potential to influence driver behavior. The objective of this study was to describe the role, formation, and lessons learned from a community advisory board (CAB) in the development and dissemination of a community-based rural roadway safety campaign. The CAB provided campaign input through quarterly meetings and email. The campaign had three main CAB and crash data-informed messages: 1) Slow Down, 2) Leave More Space, and 3) Avoid Passing. The CAB led campaign activities to publicize the message, distribute swag, and organize event logistics. To evaluate CAB effectiveness and inform future community engagement efforts, we conducted in-depth, semi-structured telephone interviews with CAB members in July 2020. Interviews were transcribed, coded, and codes were categorized into five main themes. Overall, CAB membership was described as an overwhelmingly positive experience in terms of the CAB structure, culture fostered among the group, responsibilities, and time commitment. Board members reported successful campaign implementation, gave positive feedback regarding the research team's engagement efforts, and provided valuable recommendations for future campaigns (e.g., adding social media components, expansion of CAB age and industry diversity, and increasing group bonding activities). Results from this study demonstrate the instrumental role and logistics involved in engagement of community advisors for the development and implementation of a rural roadway safety campaign. Steps and lessons from this study can be applied to other community-level injury and violence prevention topics, with a particular focus on rural communities.


SHUTTLE-STREAMING: SYNCHRONIZATION WITH HEAT PRODUCTION IN SLIME MOLD.
Two small blobs and a channel excised from a slime mold plasmodium were allowed to fuse into a dumbbell-shaped mass in a thermally insulated Kamiya double chamber equipped with naked bead thermistors in contact with the blobs. Cyclic temperature differences of from 1 X 10(-4) to 5 X 10(-2) deg Celsius were recorded by a sensitive lock-in amplifier method with a basal noise level of less than 2 X 10(-2)deg Celsius and a time constant of 0.5 second. The temperature differences were caused by periodic bursts of heat production synchronized perfectly with the shuttle-streaming cycle and invariably localized at the source rather than the destination of the streaming cytoplasm. The results support the theory that the motive force for cytoplasmic streaming in the slime mold is pressure, probably generated by contraction of elements in the channel walls.


Thyroid Function among Breastfed Children with Chronically Excessive Iodine Intakes.
Iodine excess may impair thyroid function and trigger adverse health consequences for children. This study aims to describe iodine status among breastfed infants with high iodine exposure in the Saharawi refugee camps Algeria, and further assess thyroid function and iodine status among the children three years later. In 2010, a cross-sectional study among 111 breastfed children aged 0-6 months was performed (baseline study). In 2013, a second cross-sectional study (follow-up study) was conducted among 289 children; 213 newly selected and 76 children retrieved from baseline. Urinary iodine concentration (UIC) and breast milk iodine concentration (BMIC) were measured at baseline. UIC, thyroid hormones and serum thyroglobulin (Tg) were measured at follow-up. At baseline and follow-up, 88% and 72% had excessive iodine intakes (UIC ≥ 300 µg/L), respectively. At follow-up, 24% had a thyroid hormone disturbance and/or elevated serum Tg, including 9% with subclinical hypothyroidism (SCH), 4% with elevated fT3 and 14% with elevated Tg. Children with SCH had poorer linear growth and were more likely to be underweight than the children without SCH. Excessive iodine intakes and thyroid disturbances were common among children below four years of age in our study. Further, SCH seemed to be associated with poor growth and weight.


Illness course, comorbidity, gender, and suicidality in patients with bipolar disorder.
Among patients with bipolar disorder, comorbid conditions are common. Comorbidity is associated with a more difficult course of illness (such as longer episodes, shorter time euthymic, and earlier age at onset) and an increase in related problems (such as suicidality and violence). Data from the Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD) reveal that anxiety disorders, attention-deficit/hyperactivity disorder, and substance and alcohol use disorder are conditions that commonly co-occur with bipolar disorder. This article details these findings and discusses the complications associated with these comorbid conditions. STEP-BD data about gender differences are also discussed, and correlates of suicidal ideation among patients entering the program are described.


Colposcopy and cervical biopsy educational training models.
Realistic simulation models serve meaningful educational purposes. The intent of this article is to review the physical and educational features of three instructional colposcopy models, and to discuss the advantages and implications of the use of models in teaching colposcopic skills. Models made from latex, steak, and the bovine cervix were assessed as instructional devices to simulate the uterine cervix and cervical disease. Model construction, preparation, and unique features were critiqued. The models were also evaluated for their potential use in the preclinical teaching of important colposcopic skills. The latex model required minimal assembly but was costly. It was limited in design, which would exclude the training of most colposcopic techniques. The steak model used readily available materials, facilitated colposcopic skill acquisition, and was the least expensive model. The bovine model best simulated human cervical tissue, was intermediate in cost, and enabled teaching a variety of colposcopic procedures. The optional silicone inserts realistically demonstrated the spectrum of cervical disease. Various cervical biopsy models have different strengths and weaknesses for teaching colposcopic procedural skills. Available models each possess unique features designed to reproduce anatomy, pathology, tissue texture, and technical spatial limitations. The models permit assessment of gynecologic knowledge and psychomotor abilities.


Blood and urine mercury levels in adult amalgam patients of a randomized controlled trial: interaction of Hg species in erythrocytes.
Parts of the population are permanently exposed to low levels of Hg degrees and Hg(II) from dental amalgam. It was the aim (1) to investigate the internal exposure to amalgam-related mercury from the kinetics of inorganic Hg in plasma and erythrocytes after amalgam removal, and (2) to estimate the amalgam-related absorbed dose. Dietary coexposure was monitored by determination of blood organic-Hg. Postremoval steady-state Hg concentrations were measured for 18 months. Eighty-two patients had been randomized into three groups: (A) removal of the fillings; (B) removal and non-specific detoxification, and (C) a health promotion program without removal. After amalgam removal, inorganic Hg dropped rapidly in plasma and red cells, stabilizing at 27% of preremoval levels after 60 days. Concentrations of organic Hg in plasma remained unchanged, indicating no change in dietary uptake of organic Hg. The concentration of organic Hg in red cells of group A was in the early postremoval phase lower and in the late postremoval phase higher than the preremoval control (p<0.01 for low-high difference). A protracted increase in organic Hg was also found in red cells of group B after 60 days. Thus, the effect of removal on organic Hg levels in the combined group A+B was compared with the values of group C in a linear mixed effects (LME) model which showed a significant increase with time in group A+B (p=0.028). In all groups, time profiles of urinary concentration and excretion of total-Hg were very similar to those of inorganic-Hg levels in plasma. From extrapolations of blood and urine data it was estimated that the amalgam-related inhalation and ingestion of Hg species were within the limits proposed by WHO, ATSDR and EPA. The integrated daily Hg dose absorbed from amalgam was estimated up to 3 microg for an average number of fillings and at 7.4 for a high amalgam load. This is the first study on adult amalgam patients which continuously monitored the postremoval decline of inorganic Hg and the coexposure from dietary organic Hg in a randomized-controlled-trial design. The integrated daily dose of 7.4 microg absorbed from a high amalgam load is well below the tolerable dose of 30 microg (WHO, 1990). The unexpected postremoval increase in erythrocyte organic Hg, which is associated with the depletion of cellular inorganic Hg, might result from binding of organic Hg to cellular sites previously occupied by inorganic Hg.


Assessing the impact of introductory programming workshops on the computational reproducibility of biomedical workflows.
As biomedical research becomes more data-intensive, computational reproducibility is a growing area of importance. Unfortunately, many biomedical researchers have not received formal computational training and often struggle to produce results that can be reproduced using the same data, code, and methods. Programming workshops can be a tool to teach new computational methods, but it is not always clear whether researchers are able to use their new skills to make their work more computationally reproducible. This mixed methods study consisted of in-depth interviews with 14 biomedical researchers before and after participation in an introductory programming workshop. During the interviews, participants described their research workflows and responded to a quantitative checklist measuring reproducible behaviors. The interview data was analyzed using a thematic analysis approach, and the pre and post workshop checklist scores were compared to assess the impact of the workshop on the computational reproducibility of the researchers' workflows. Pre and post scores on a checklist of reproducible behaviors did not change in a statistically significant manner. The qualitative interviews revealed that several participants had made small changes to their workflows including switching to open source programming languages for their data cleaning, analysis, and visualization. Overall many of the participants indicated higher levels of programming literacy, and an interest in further training. Factors that enabled change included supportive environments and an immediate research need, while barriers included collaborators that were resistant to new tools, and a lack of time. While none of the workshop participants completely changed their workflows, many of them did incorporate new practices, tools, or methods that helped make their work more reproducible and transparent to other researchers. This indicates that programming workshops now offered by libraries and other organizations contribute to computational reproducibility training for researchers.


Exploring Fatigue Trajectories in Early Symptomatic Knee and Hip Osteoarthritis: 6-year Results from the CHECK Study.
To examine whether different groups of fatigue trajectories can be identified among patients with early symptomatic osteoarthritis (OA) of the knee or hip, to describe the level of fatigue severity within each of these fatigue groups, and to investigate the involvement of age, sex, use of medication, comorbidity, and OA severity in relation to group membership. Six years of followup data on fatigue (Medical Outcomes Study Short Form-36 Vitality scale) came from the Cohort Hip and Cohort Knee (CHECK) cohort. Growth mixture modeling was applied to identify distinct fatigue trajectories as well as to take into account the effects of the patient characteristics. Three fatigue trajectories were identified: low fatigue, low-to-high fatigue, and high fatigue. Latter trajectories showed considerable overlap from years 2 to 6, but differed in some patient characteristics in comparison with each other and in comparison with the low fatigue group. Comorbidity, medication use, and sex were significantly associated with the identified trajectories. Women, individuals with a comorbid disease, and those who used medication were more likely to follow a high fatigue trajectory. These findings suggest heterogeneous development of fatigue in the early OA population associated with varying patient characteristics. Further, this study shows that a considerable number of patients with OA already experience elevated levels of fatigue at an early stage of OA. While these findings need to be replicated, the identification of these trajectories with differing patient characteristics may warrant tailored psychosocial interventions for patients with elevated levels of fatigue.


Evolution and resolution of oedema following severe temporary cerebral ischaemia in the gerbil.
Regional cerebral blood flow (rCBF) and oedema following profound temporary ischaemia were studied in the gerbil. Ninety-four per cent of animals died within 24 hours of reperfusion; 50% by 4 hours. Regional differences in oedema (specific gravity method), Evans blue (EB) staining and rCBF (hydrogen clearance technique) occurred. Oedema developed during arterial occlusion, being inversely proportional to residual flow and was markedly exacerbated during reperfusion. Reperfusion hyperaemia was maximal in the parietal and hippocampal regions (ischaemic rCBF 4 ml 100 g-1 min-1). Oedema was disappearing in all areas by 3 hours of reperfusion and autoregulation returned in the occipital region (mean ischaemia rCBF 8 ml 100 g-1 min-1). EB staining and haemorrhage appeared in the thalamus (rCBF 10 ml 100 g-1 min-1) as oedema was decreasing. It is suggested that the amount of oedema and hyperaemia during reperfusion are dependent on the severity of the ischaemia. Areas of moderate ischaemia (8-10 ml 100 g-1 min-1) show little hyperaemia and greater oedema resolution during reperfusion as compared to areas of severe ischaemia (circa 4 ml 100 g-1 min-1) where there is marked hyperaemia with less oedema resolution. Early in the reperfusion period, oedema is not associated with EB staining and indicates a cytotoxic mechanism. The vasogenic component, with macroscopic haemorrhages and leakage of EB occurs later. In this model it is concluded that the early cytotoxic oedema formation and hyperaemia are phenomena with little bearing on mortality, which correlates better with later vasogenic changes.


Critical analysis of the Chiari malformation Type I found in children with lipomyelomeningocele.
Occasional comments are found in the literature regarding patients with lipomyelomeningocele and concomitant Chiari malformation Type I (CM-I). The object of this study was to explore the association between these two conditions. The authors performed a retrospective database analysis of lipomyelomeningocele cases to identify cases of concomitant CM-I. Analysis of posterior fossa volume (based on the Cavalieri principle) was performed in all identified cases in which appropriate neuroimages were available, and the results were compared with those obtained in age-matched controls. Seven (13%) of 54 patients with lipomyelomeningocele were found to also have CM-I. Two of these were symptomatic (cervicothoracic syrinx and occipital headaches) and required posterior fossa decompression. No correlation was found between the amount of hindbrain herniation and the level of the conus medullaris or the type of lipomyelome-ningocele (for example, caudal or transitional). Volumetric studies of the posterior fossa revealed normal age-matched volumes in all but one patient (who had asymptomatic CM-I). The incidence of CM-I in patients with lipomyelomeningocele appears to be significantly greater than that of the general population and the association rate is too high for the finding to be a chance occurrence. Decreases in the volume of the posterior cranial fossa were not found in the majority of patients in this small cohort; therefore, the cause of the concomitant occurrence of lipomyelomeningocele and CM-I remains undetermined. Clinicians should consider obtaining imaging studies of the entire neuraxis in patients with lipomyelomeningoceles and should investigate other causes for syringes found in association with lipomyelomeningoceles.


Impact of enterococcal colonization and infection in solid organ transplantation recipients from the Swiss transplant cohort study.
The burden of enterococcal infections has increased over the last decades with vancomycin-resistant enterococci (VRE) being a major health problem. Solid organ transplantation is considered as a risk factor. However, little is known about the relevance of enterococci in solid organ transplantation recipients in areas with a low VRE prevalence. We examined the epidemiology of enterococcal events in patients followed in the Swiss Transplant Cohort Study between May 2008 and September 2011 and analyzed risk factors for infection, aminopenicillin resistance, treatment, and outcome. Of the 1234 patients, 255 (20.7%) suffered from 392 enterococcal events (185 [47.2%] infections, 205 [52.3%] colonizations, and 2 events with missing clinical information). Only 2 isolates were VRE. The highest infection rates were found early after liver transplantation (0.24/person-year) consisting in 58.6% of Enterococcus faecium. The highest colonization rates were documented in lung transplant recipients (0.33/person-year), with 46.5% E. faecium. Age, prophylaxis with a betalactam antibiotic, and liver transplantation were significantly associated with infection. Previous antibiotic treatment, intensive care unit stay, and lung transplantation were associated with aminopenicillin resistance. Only 4/205 (2%) colonization events led to an infection. Adequate treatment did not affect microbiological clearance rates. Overall mortality was 8%; no deaths were attributable to enterococcal events. Enterococcal colonizations and infections are frequent in transplant recipients. Progression from colonization to infection is rare. Therefore, antibiotic treatment should be used restrictively in colonization. No increased mortality because of enterococcal infection was noted.


Nanowires network for biomolecular detection using contactless impedance tomoscopy technique.
Here we present the detection of ultralow concentrations of biomolecules in a device made from a polycarbonate membrane containing a network of gold nanowires and using a "contactless" impedance tomoscopy technique. The sensor comprises a thin dielectric layer with two parallel band electrodes on the one side and a microchannel containing gold nanowires onto which the adsorption of antibodies occurs. Upon applying a high-frequency ac voltage between the two electrodes, the adsorption process occurring at the surface of the gold nanowires can be followed through contactless impedance measurements. The configuration allows the real-time detection of biomolecules with a bulk concentration in the picomolar range.


Seedling stage strategies as a means of habitat specialization in herbaceous plants.
The regeneration niche has been little investigated in studies of community assembly and plant distribution. We examined adaptive associations between seedling traits and habitat specialization. Two habitat contrasts were investigated across several evolutionary lineages of angiosperms: species specialized to forest vs. open habitats and to dry vs. wet habitats. We also tested whether effects of shade and drought vary independently or, alternatively, if shade may amplify effects on drought-stressed plants. Seedling response in terms of growth rate, height, slenderness, specific leaf area (SLA) and degree of elongation (longest internode; petiole or leaf-sheath depending on species' morphology) to light and watering treatments was assessed. We used a factorial design involving three light regimes and two watering frequencies. The open-shaded habitat contrast and the dry-wet habitat contrast were investigated using six and five pairs of congeneric species, respectively. The congeneric species pair design controlled for confounding effects of evolutionary history prior to divergence in habitat specialization. Seedling growth rate generally decreased with shade and reduced watering frequency. Plant height was generally largest at intermediate light. Specialization to shaded habitats was associated with a more conservative growth strategy, i.e. showing a more modest growth response to increasing light. Species from all habitats showed the highest relative elongation at intermediate light, except for the moist-habitat species, for which elongation increased with shade. Contrary to our expectations, species from dry habitats grew bigger than species from moist habitats in all treatments. SLA responded to the light treatment, but not to watering regime. The contrasting light and moisture conditions across habitats appear to not have selected for differences in SLA. We conclude that seedling phase strategies of resource allocation in temperate herbs contribute to their habitat specialization. Habitat-specific seedling strategies and trade-offs in response to resource availability and environmental conditions may be important to adaptive specialization.


Type of activity pacing instruction affects physical activity variability in adults with symptomatic knee or hip osteoarthritis.
In a previous pilot study, the effect of 2 types of activity pacing instruction, general versus tailored, on osteoarthritis symptoms was examined and fatigue improved in the tailored group. Because activity pacing involves instruction on physical activity engagement, we undertook this secondary analysis to examine how pacing instruction affected physical activity patterns. Thirty-two adults with knee or hip osteoarthritis, stratified by age and gender, received either tailored or general activity pacing instruction. All participants wore an accelerometer for 5 days that measured physical activity and allowed for repeated symptom assessment at baseline and 10-week follow-up. Activity patterns were assessed by examining physical activity variability (standard deviation of 5-day average activity counts per minute), and average activity level (5-day average activity counts per minute). Physical activity variability decreased in the tailored group and increased in the general group. No significant group changes in average activity from baseline to 10-week follow-up were found. In this pilot study, type of activity pacing instruction affected objective physical activity patterns in adults with OA. Tailored activity pacing was more effective at reducing high and low activity bouts corresponding to the message of keeping a steady pace to reduce symptoms.


Method Validation for the Quantitative Analysis of Aflatoxins (B1, B2, G1, and G2) and Ochratoxin A in Processed Cereal-Based Foods by HPLC with Fluorescence Detection.
Modified AOAC 991.31 and AOAC 2000.03 methods for the simultaneous determination of total aflatoxins (AFs), aflatoxin B1, and ochratoxin A (OTA) in processed cereal-based foods by RP-HPLC coupled with fluorescence detection were validated. A KOBRA® Cell derivatization system was used to analyze total AFs. One of the modifications was the extraction procedure of mycotoxins. Both AFs and OTA were extracted with methanol-water (75+25, v/v) and purified with an immunoaffinity column before HPLC analysis. The modified methods were validated by measuring the specificity, selectivity, linearity, sensitivity, accuracy, repeatability, reproducibility, recovery, LOD, and LOQ parameters. The validated methods were successfully applied for the simultaneous determination of mycotoxins in 81 processed cereal-based foods purchased in Turkey. These rapid, sensitive, simple, and validated methods are suitable for the simultaneous determination of AFs and OTA in the processed cereal-based foods.


2, 3, 5, 4'-Tetrahydroxystilbene-2-O-beta-D-glucoside improves gastrointestinal motility disorders in STZ-induced diabetic mice.
Oxidative stress has recently been considered as a pivotal player in the pathogenesis of diabetic gastrointestinal dysfunction. We therefore investigated the role of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucoside (THSG) that has a strong anti-oxidant property, in diabetic gastrointestinal dysmotility as well as the underlying protective mechanisms. THSG restored the delayed gastric emptying and the increased intestinal transit in streptozotocin (STZ)-induced diabetic mice. Loss of neuronal nitric oxide synthase (nNOS) expression and impaired nonadrenergic, noncholinergic (NANC) relaxations in diabetic mice were relieved by long-term preventive treatment with THSG. Meanwhile, THSG (10(-7)~10(-4) mol/L) enhanced concentration-dependently NANC relaxations of isolated colons in diabetic mice. Diabetic mice displayed a significant increase in Malondialdehyde (MDA) level and decrease in the activity of glutathione peroxidase (GSH-Px), which were ameliorated by THSG. Inhibition of caspase-3 and activation of ERK phosphorylation related MAPK pathway were involved in prevention of enhanced apoptosis in diabetes afforded by THSG. Moreover, THSG prevented the significant decrease in PPAR-γ and SIRT1 expression in diabetic ileum. Our study indicates that THSG improves diabetic gastrointestinal disorders through activation of MAPK pathway and upregulation of PPAR-γ and SIRT1.


In vitro fermentability of sugar beet pulp derived oligosaccharides using human and pig fecal inocula.
The in vitro fermentation characteristics of different classes of sugar beet pectic oligosaccharides (SBPOS) were studied using human and pig fecal inocula. The SBPOS consisted mainly of partially acetylated rhamnogalacturonan oligosaccharides and partially methyl-esterified/acetylated homogalacturonan oligosaccharides. Some SBPOS contained an unsaturated galacturonic acid residue at their non-reducing end. It was shown that SBPOS could be completely fermented by human and pig fecal microbiota, thereby producing butyrate yet mainly acetate and propionate as metabolites. The degradation of SBPOS by pig fecal microbiota was different and much slower compared to human fecal microbiota. In general, rhamnogalacturonan oligosaccharides were degraded slower than homogalacturonan oligosaccharides. Acetylation of rhamnogalacturonan oligosaccharides lowered the degradation rate by pig fecal microbiota but not by human fecal microbiota. No classic bifidogenic effect was shown for SBPOS using human fecal inoculum. However, several other potentially interesting modifications in the microbiota composition that can be associated with host health were observed, which are discussed.


A three-step least-squares iterative method for tilt phase-shift interferometry.
An iterative method based on least-squares fittings is proposed to retrieve wavefront phase from tilt phase-shift interferograms. In each iteration cycle, proposed method calculates wavefront phase and tilt phase shifts in x- and y-directions in three individual least-squares fitting steps. In tilt phase shifts extracting steps, phase shifts of interferograms columns or rows are calculated with least-squares method, and then tilt phase shifts of interferograms in x- or y-direction are determined by linear regressions. At least three interferograms of three by three pixels are required with proposed method. The performance of proposed method is demonstrated by simulations and experiments. Tilt gradients and translational phase shifts could be extracted with high accuracy and large wavefront tilts could be well handled with proposed method. The method could be applied to temporal phase-shift interferometers with uncalibrated transducers or that in vibrating environment.


Tie2-expressing monocytes and tumor angiogenesis: regulation by hypoxia and angiopoietin-2.
Recent findings indicate that tumor-associated macrophages are important drivers of tumor angiogenesis. Here, we review the essential role played by Tie2-expressing monocytes (TEM) in this phenomenon. TEMs are present in human blood and tumors and their elimination in various tumor models suppresses tumor angiogenesis. A ligand for Tie2, angiopoietin-2 (Ang-2), is produced by angiogenic tumor vessels and is a chemoattractant for TEMs. Hypoxia up-regulates Tie2 expression on TEMs and, together with Ang-2, down-regulates their antitumor functions. Learning more about the regulation of TEMs by the tumor microenvironment may yield new strategies to ablate the tumor vasculature.


An Alternative Medical Diagnosis Method: Biosensors for Virus Detection.
Infectious diseases still pose an omnipresent threat to global and public health, especially in many countries and rural areas of cities. Underlying reasons of such serious maladies can be summarized as the paucity of appropriate analysis methods and subsequent treatment strategies due to the limited access of centralized and equipped health care facilities for diagnosis. Biosensors hold great impact to turn our current analytical methods into diagnostic strategies by restructuring their sensing module for the detection of biomolecules, especially nano-sized objects such as protein biomarkers and viruses. Unquestionably, current sensing platforms require continuous updates to address growing challenges in the diagnosis of viruses as viruses change quickly and spread largely from person-to-person, indicating the urgency of early diagnosis. Some of the challenges can be classified in biological barriers (specificity, low number of targets, and biological matrices) and technological limitations (detection limit, linear dynamic range, stability, and reliability), as well as economical aspects that limit their implementation into resource-scarce settings. In this review, the principle and types of biosensors and their applications in the diagnosis of distinct infectious diseases were comprehensively explained. The deployment of current biosensors into resource-scarce settings is further discussed for virus detection by elaborating the pros and cons of existing methods as a conclusion and future perspective.


Mutation analysis of HPS1, the gene mutated in Hermansky-Pudlak syndrome, in patients with isolated platelet dense-granule deficiency.
Isolated platelet dense granule (PDG) deficiency is a heterogeneous disorder frequently found among patients with mild to moderate bleeding diatheses. However, the molecular basis of this disorder is unknown. Genes involved in other rare bleeding disorders with associated reduction in the numbers of platelet dense-granules may play a role in isolated PDG deficiency. Among such genes, HPS1 is known to play a key role in the genesis of PDG and as many as 18 different HPS1 mutations have been identified in patients with Hermansky-Pudlak syndrome. Recently, we have identified subjects with one HPS1 heterozygous mutation displaying significant reductions in PDG without the clinical phenotype of Hermansky-Pudlak syndrome. This suggested that HPS1 mutations could be involved in isolated PDG deficiency. We sequenced all coding exons, and flanking intron regions of HPS1 in 16 patients with mild to severe PDG deficiency, most of whom had mild bleeding episodes. Nine patients reported a familial history of bleeding diathesis with PDG deficiency. We also evaluated the prevalence of HPS1 variations in 215 controls. Transmission electron microscopy was used to evaluate the number and morphology of PDG from patients and selected controls. No patient with PDG deficiency carried severe mutations of the HPS1 gene. We identified 6 previously described and 5 new polymorphisms in the HPS1 gene. Platelet electron microscopy in controls carrying these polymorphisms revealed that they did not significantly modify the number or morphology of PDG. Mutations affecting the HPS1 gene play a minor role in isolated PDG deficiency. These results support a molecular heterogeneity responsible for the number and morphology of PDG.


Deleterious GRM1 mutations in schizophrenia.
We analysed a phenotypically well-characterised sample of 450 schziophrenia patients and 605 controls for rare non-synonymous single nucleotide polymorphisms (nsSNPs) in the GRM1 gene, their functional effects and family segregation. GRM1 encodes the metabotropic glutamate receptor 1 (mGluR1), whose documented role as a modulator of neuronal signalling and synaptic plasticity makes it a plausible schizophrenia candidate. In a recent study, this gene was shown to harbour a cluster of deleterious nsSNPs within a functionally important domain of the receptor, in patients with schizophrenia and bipolar disorder. Our Sanger sequencing of the GRM1 coding regions detected equal numbers of nsSNPs in cases and controls, however the two groups differed in terms of the potential effects of the variants on receptor function: 6/6 case-specific and only 1/6 control-specific nsSNPs were predicted to be deleterious. Our in-vitro experimental follow-up of the case-specific mutants showed that 4/6 led to significantly reduced inositol phosphate production, indicating impaired function of the major mGluR1 signalling pathway; 1/6 had reduced cell membrane expression; inconclusive results were obtained in 1/6. Family segregation analysis indicated that these deleterious nsSNPs were inherited. Interestingly, four of the families were affected by multiple neuropsychiatric conditions, not limited to schizophrenia, and the mutations were detected in relatives with schizophrenia, depression and anxiety, drug and alcohol dependence, and epilepsy. Our findings suggest a possible mGluR1 contribution to diverse psychiatric conditions, supporting the modulatory role of the receptor in such conditions as proposed previously on the basis of in vitro experiments and animal studies.


Nanogenerators based on vertically aligned InN nanowires.
Piezoelectric nanogenerators (NGs) based on vertically aligned InN nanowires (NWs) are fabricated, characterized, and evaluated. In these NGs, arrays of p-type and intrinsic InN NWs prepared by plasma-assisted molecular beam epitaxy (MBE) demonstrate similar piezoelectric properties. The p-type NGs show 160% more output current and 70% more output power product than the intrinsic NGs. The features driving performance enhancement are reduced electrostatic losses due to better NW array morphology, improved electromechanical energy conversion efficiency due to smaller NW diameters, and the higher impedance of intrinsic NGs due to elevated NW surface charge levels. These findings highlight the potential of InN based NGs as a power source for self-powered systems and the importance of NW morphology and surface state in overall NG performance.


Multinational evidence-based recommendations for the diagnosis and management of gout: integrating systematic literature review and expert opinion of a broad panel of rheumatologists in the 3e initiative.
We aimed to develop evidence-based multinational recommendations for the diagnosis and management of gout. Using a formal voting process, a panel of 78 international rheumatologists developed 10 key clinical questions pertinent to the diagnosis and management of gout. Each question was investigated with a systematic literature review. Medline, Embase, Cochrane CENTRAL and abstracts from 2010-2011 European League Against Rheumatism and American College of Rheumatology meetings were searched in each review. Relevant studies were independently reviewed by two individuals for data extraction and synthesis and risk of bias assessment. Using this evidence, rheumatologists from 14 countries (Europe, South America and Australasia) developed national recommendations. After rounds of discussion and voting, multinational recommendations were formulated. Each recommendation was graded according to the level of evidence. Agreement and potential impact on clinical practice were assessed. Combining evidence and clinical expertise, 10 recommendations were produced. One recommendation referred to the diagnosis of gout, two referred to cardiovascular and renal comorbidities, six focused on different aspects of the management of gout (including drug treatment and monitoring), and the last recommendation referred to the management of asymptomatic hyperuricaemia. The level of agreement with the recommendations ranged from 8.1 to 9.2 (mean 8.7) on a 1-10 scale, with 10 representing full agreement. Ten recommendations on the diagnosis and management of gout were established. They are evidence-based and supported by a large panel of rheumatologists from 14 countries, enhancing their utility in clinical practice.


The ventromedial hypothalamic area and the vagus are neural substrates for anticipatory insulin release.
It has been clearly demonstrated in a number of animal species including man, that insulin secretion can occur in response to sensory stimuli associated with meal taking and in advance of any ingestional rise in blood nutrients. The neural basis of this anticipatory insulin rise is poorly delineated and was therefore investigated in two experiments. In the first, rats with electrolytic lesions of the ventromedial hypothalamic area (VMH), or sham lesions were assessed and in the second experiment rats with bilateral subdiaphragmatic vagotomies or sham vagotomies were studied. In both experiments rats with jugular cannulae were conditioned to expect a meal 5 min following the beginning of a complex (light, tone and odor) stimulus. On the test trial the stimulus was given but no meal was forthcoming. Blood was sampled via the jugular catheter before and during the stimulus presentation. In both experiments the control rats showed a significant insulin rise during the stimulus presentation. However, neither the VMH-lesioned nor the vagotomized rats showed any significant change in insulin levels. The results clearly demonstrate an insulin rise to a complex sensory stimulus which has been paired with food and further show the dependence of that response on the integrity of the hypothalamic-vagal-visceral neural pathway. It is suggested that the VMH receives meal-associated sensory input and employs that information to prepare the animal metabolically for incoming nutrients. One aspect of this anticipatory mechanism is a vagally mediated insulin secretion.


Complications after bronchoscopic lung volume reduction.
Bronchoscopic lung volume reduction (BLVR) has been demonstrated an efficient and safe alternative to surgery in multiple randomized trials and retrospective cohort studies. However, despite its minimal invasiveness BLVR is not without potential harm. Complications and their incidents differ significantly between the individual BLVR techniques (valves, coils, vapour or sealant) which are bearing varying device- and intervention-dependent risks. Interventional pulmonologists must be aware of potential side effects and their management to anticipate and ensure highest quality treatment of the severely ill emphysema patient.