Prevalence and Trends of Metabolic Syndrome in Slovakia during the Period of 2003-2012.
Metabolic syndrome is a combination of clinical risk factors for cardiovascular disease as well as for diabetes. Metabolic syndrome arises from insulin resistance accompanied with abnormal adipose deposition. The aim of our cross-sectional time trends study was to characterize the prevalence of metabolic syndrome and its five risk determinants among the clients of Health Advice Centres of Regional Public Health Authorities in Slovakia. The study was stratified by gender and age groups during the 10 year period from 2003–2012. Prevalence data were estimated in adults and children (≥10 years, N=79,904) from the nationwide electronic database of Health Advice Centres of Regional Public Health Authorities in Slovak Republic "Test of healthy heart" from 2003 to 2012. The overall prevalence of metabolic syndrome was 30.2% in males and 26.6% in females, abdominal obesity was confirmed in 48.3% of males and 53.9% of females. Increased triglyceride level has higher prevalence among males (33.3%) compared to females (24.2%). Blood pressure (BP) values and fasting glucose values were significantly higher in males (58.2%) than females (41.9%). During the 10 year period from 2003 to 2012, we confirmed an increased trend in the age-adjusted prevalence of metabolic syndrome. Abdominal obesity and elevated triglycerides had also increased time trends prevalence in both sexes. The prevalence of people without risk determinants of metabolic syndrome had a time decreasing trend. A surprising finding is a decrease in the proportion of persons with suboptimal HDL-cholesterol. The proportion of people with elevated BP and glucose showed little change during the reporting period. The increasing prevalence of metabolic syndrome, abdominal obesity, and elevated triglycerides highlights the urgency of addressing these health problems as a healthcare priority to reduce cardiovascular mortality in the Slovak Republic.


Preferential loss of the paternal alleles in the 18q- syndrome.
Individuals with the 18q- syndrome have variable deletions from the long arm of chromosome 18. They also exhibit a highly variable phenotype. To correlate genotype with phenotype accurately, extensive molecular and phenotypic analyses are needed on each affected individual. As a part of this analysis, we have determined the parental origin of the deleted chromosome in 34 individuals with the 18q- syndrome. We have found that 85% of the de novo deletions are paternal in origin. The percentage of fathers of individuals with paternally derived deletions who were > 30 years old was (not significantly) greater than that of the general population. The mothers of individuals with maternally derived deletions were near an average age for childbearing compared to the general population. Individuals with maternally derived terminal deletions had breakpoints as varied as those with paternally derived deletions. These results are consistent with the hypothesis that the reduced incidence of maternally derived deletions is not due to reduced viability, since individuals with large maternally derived deletions of chromosome 18q were found. We hypothesize that the prevalence of paternally derived deletions is due to an increased frequency of chromosome breakage in male germ cells. These results are consistent with results observed in other segmental aneusomies in which there is a high incidence of paternally derived deletions.


Antioxidants protect against glutamate-induced cytotoxicity in a neuronal cell line.
The effects of reducing agents and antioxidants on L-Glutamate (Glu)-induced cytotoxicity were examined in the N18-RE-105 neuronal cell line. The cytotoxicity by Glu (1 and 10mM) was potentiated by exposure to growth medium containing a low concentration of cystine (5-100 microM), instead of the normal medium containing 200 microM cystine. In contrast, the toxicity was suppressed by increasing the cystine concentration to 500 to 1000 microM. Reducing agents, cysteine (30-1000 microM), dithiothreitol (10-250 microM) and glutathione (GSH, 10-1000 microM) also protected the cells against the cytotoxicity of 10 mM Glu in a concentration-dependent manner. The antioxidants vitamin E (10-100 microM), idebenone (0.1-3 microM) and vinpocetine (10-100 microM) also provided marked protection against the cytotoxicity of Glu (10 mM) or quisqualate (1 mM). Antioxidants also prevented the delayed cell death caused by lowering the concentration of cystine in the medium to 5 microM. Incubation of the cells with 10 mM Glu caused a marked decrease in cellular GSH levels. Although cysteine and dithiothreitol prevented the GSH reduction caused by Glu, antioxidants did not. The cellular levels of oxidants were assessed using 2,7-dichlorofluorescin, a probe that accumulates within cells and is converted to a fluorescent product by oxidation. Glu (10 mM) caused a marked increase in such fluorescence, whereas vitamin E and idebenone reduced markedly the number of fluorescent cells to control levels even added with 10 mM Glu. These results indicate that oxidative stress due to loss of cellular levels of GSH is one mechanism whereby Glu/quisqualate exert cytotoxicity and suggest that centrally active antioxidants may reduce neuronal damage in pathologic conditions associated with excessive Glu release.


Serotyping of dengue virus in 2016-17 outbreak in Kudat, Sabah, Malaysia.
An outbreak of dengue in Kudat, northern Sabah in 2016-2017 provided an opportunity to investigate the circulating serotypes of dengue viruses of cases at Hospital Kudat. Between September 2016 and December 2017, a total of 156 dengue positive sera (tested positive by either NS1 antigen, or IgM and IgG antibody rapid test) were collected from dengue patients who had acute fever and showed signs and symptoms suggestive of dengue. RNA was extracted from the sera using QIAamp RNA Blood Mini Kit, and molecular amplification was performed using one-step RT-PCR kit, followed by nested PCR using HotStart Taq master mix kit with the primers of the dengue C-prM gene. There were 81 (52%) male and 75 (48%) female cases. The age group with the highest number of cases was the 10-19 years old, while the youngest infected was 8 months old and the oldest was 83 years old. RT-PCR results showed 88 sera dengue positive, 48 infected with a single serotype while another 40 with multiple serotypes. All four DENV serotypes were co-circulating during the outbreak period and DENV-1 was predominant. Molecular analysis also indicated 69.2%, 50.0%, 51.9% and 48.9% respectively of the NS1, IgM, IgG and IgM and IgG positive sera were RT-PCR positive for dengue. High number of cases were seen in December 2016, February and May 2017. The dengue outbreak might be related to switching of predominant serotype from DENV 4 to DENV 1.


Increased Antifungal Drug Resistance in Clinical Isolates of Cryptococcus neoformans in Uganda.
Cryptococcal antigen screening is recommended among people living with AIDS when entering HIV care with a CD4 count of <100 cells/μl, and preemptive fluconazole monotherapy treatment is recommended for those with subclinical cryptococcal antigenemia. Yet, knowledge is limited of current antimicrobial resistance in Africa. We examined antifungal drug susceptibility in 198 clinical isolates collected from Kampala, Uganda, between 2010 and 2014 using the CLSI broth microdilution assay. In comparison with two previous studies from 1998 to 1999 that reported an MIC50 of 4 μg/ml and an MIC90 of 8 μg/ml prior to widespread human fluconazole and agricultural azole fungicide usage, we report an upward shift in the fluconazole MIC50 to 8 μg/ml and an MIC90 value of 32 μg/ml, with 31% of isolates with a fluconazole MIC of ≥ 16 μg/ml. We observed an amphotericin B MIC50 of 0.5 μg/ml and an MIC90 of 1 μg/ml, of which 99.5% of isolates (197 of 198 isolates) were still susceptible. No correlation between MIC and clinical outcome was observed in the context of amphotericin B and fluconazole combination induction therapy. We also analyzed Cryptococcus susceptibility to sertraline, with an MIC50 of 4 μg/ml, suggesting that sertraline is a promising oral, low-cost, available, novel medication and a possible alternative to fluconazole. Although the CLSI broth microdilution assay is ideal to standardize results, limit human bias, and increase assay capacity, such assays are often inaccessible in low-income countries. Thus, we also developed and validated an assay that could easily be implemented in a resource-limited setting, with similar susceptibility results (P = 0.52).


Listeriosis: a model for the fine balance between immunity and morbidity.
Listeriosis is a severe food-borne disease caused by Listeria monocytogenes. It mostly affects immune-compromised individuals, pregnant women, and the elderly, and it is associated with huge economic losses, especially to the food industry. In the last decade, a sharp increase in listeriosis incidence was observed in several European countries. No suitable explanation was found for this increase, which occurred only in old patients and not in pregnant women. We developed a mathematical model to explore this upsurge by studying the balance between the immunized population fraction and the force of infection, and its influence on the incidence of listeriosis. The model shows that the current upsurge could be the result of a decrease in exposure to the pathogen in food a few decades ago and hence decreased level of population immunity. The model also suggests that, counterintuitively, the incidence of listeriosis can be higher under reduced exposure to L. monocytogenes than under high exposure. These results rely on the accepted assumption that immunity to L. monocytogenes is long-lived (at least 20 years) or that there is a long-lived boosting effect by previous exposure to L. monocytogenes. The results are robust to wide changes in all other model parameters. Historical alterations in exposure to L. monocytogenes might explain current changes in incidence of listeriosis. The model may also be implied for other noncontagious infectious diseases (eg, food borne diseases or vector-borne diseases for which humans are considered dead-end hosts) for which susceptibility increases with age.


Oxygen isotope analysis of bacterial and fungal manganese oxidation.
The ability of micro-organisms to oxidize manganese (Mn) from Mn(II) to Mn(III/IV) oxides transcends boundaries of biological clade or domain. Many bacteria and fungi oxidize Mn(II) to Mn(III/IV) oxides directly through enzymatic activity or indirectly through the production of reactive oxygen species. Here, we determine the oxygen isotope fractionation factors associated with Mn(II) oxidation via various biotic (bacteria and fungi) and abiotic Mn(II) reaction pathways. As oxygen in Mn(III/IV) oxides may be derived from precursor water and molecular oxygen, we use a twofold approach to determine the isotope fractionation with respect to each oxygen source. Using both 18 O-labeled water and closed-system Rayleigh distillation approaches, we constrain the kinetic isotope fractionation factors associated with O atom incorporation during Mn(II) oxidation to -17.3‰ to -25.9‰ for O2 and -1.9‰ to +1.8‰ for water. Results demonstrate that stable oxygen isotopes of Mn(III/IV) oxides have potential to distinguish between two main classes of biotic Mn(II) oxidation: direct enzymatic oxidation in which O2 is the oxidant and indirect enzymatic oxidation in which superoxide is the oxidant. The fraction of Mn(III/IV) oxide-associated oxygen derived from water varies significantly (38%-62%) among these bio-oxides with only weak relationship to Mn oxidation state, suggesting Mn(III) disproportionation may account for differences in the fraction of mineral-bound oxygen from water and O2 . Additionally, direct incorporation of molecular O2 suggests that Mn(III/IV) oxides contain a yet untapped proxy of δ18OO2 of environmental O2 , a parameter reflecting the integrated influence of global respiration, photorespiration, and several other biogeochemical reactions of global significance.


Management of Large Proximal Ureteral Stones: A Comparative Clinical Trial Between Transureteral Lithotripsy (TUL) and Shock Wave Lithotripsy (SWL).
A review of the related medical journals indicates that there is no definite evidence-based option for managing large proximal ureteral stones, although many procedures such as transureteral lithotripsy (TUL), shock wave lithotripsy (SWL), percutaneous nephrolithotripsy, laparoscopic ureterolithotomy, and open ureterolithotomy are currently used to treat this urological problem. In this study, we tried to determine the most appropriate treatment plan for proximal ureteral stones larger than 12 mm by comparing the two most commonly used procedures. Between February 2005 and April 2011, 62 patients including 40 males and 22 females (mean age 39.5 years, range 19 to 64) with proximal ureteral stones larger than 12 mm (12-26 mm) with a mean size of 17.64 mm were prospectively divided into two groups consisting of 32 patients who underwent TUL (group A) and 30 who underwent SWL (group B). In unsuccessful cases, repeat SWL or TUL was planned. Patients who could not tolerate the lithotomy position, younger than 18 years, had undergone coagulopathy, had concurrent renal and ureteral stones, were pregnant, or had sepsis were excluded from this study. Stone access was successful in 28 patients and the treatment was efficient in 18 patients (56.25%) in group A. For the patients with successful stone access but unsuccessful TUL, a DJ was inserted and a second ureteroscopic intervention was performed. The second intervention was successful in 7 patients (21.87). SWL was successful in 14 patients (46.66%) in the first attempt and in 7 additional patients in the second intervention (23.33%). In this study, we showed different success rates for SWL and TUL because of the larger size of the stones. We achieved a success rate of 56.25% in the first attempt in the TUL group, and the overall success rate (after the second TUL) was 78.12%. In comparison, the SWL group had a success rate of 46.66% in the first attempt, and the overall success rate (after the second SWL) was 69.96%.


Curcumin analog GO-Y030 inhibits tumor metastasis and glycolysis.
Tumor metastasis is one of the worst prognostic features of cancer. Although metastasis is a major cause of cancer-related deaths, an effective treatment has not yet been established. Here, we explore the anti-tumor effects of GO-Y030, a curcumin analog, via various mechanisms using a mouse model. GO-Y030 treatment of B16-F10 melanoma cells inhibited TGF-β expression and glycolysis. The invasion assay results showed almost complete invasion inhibition following GO-Y030 treatment. Mouse experiments demonstrated that GO-Y030 administration inhibited lung tumor metastasis without affecting vascular endothelial cells. Consistent with this result, GO-Y030 treatment led to the downregulation of MMP2 and VEGFα, inhibiting tumor invasion and metastasis. The silencing of eIF4B, a downstream molecule of S6, attenuated MMP2 expression. Our study demonstrates the novel efficacy of GO-Y030 in inhibiting tumor metastasis by regulating metastasis-associated gene expression via inhibiting dual access, glycolytic, and TGF-β pathways.


The Association between Pediatric NAFLD and Common Genetic Variants.
Non-alcoholic fatty liver disease (NAFLD) is one of the most common complications of obesity. Several studies have shown that genetic predisposition probably plays an important role in its pathogenesis. In fact, in the last few years a large number of genetic studies have provided compelling evidence that some gene variants, especially those in genes encoding proteins regulating lipid metabolism, are associated with intra-hepatic fat accumulation. Here we provide a comprehensive review of the gene variants that have affected the natural history of the disease.


The anti-angiogenic agent lenvatinib induces tumor vessel normalization and enhances radiosensitivity in hepatocellular tumors.
The evaluation of angiogenesis inhibitors requires the analysis of the precise structure and function of tumor vessels. The anti-angiogenic agents lenvatinib and sorafenib are multi-target tyrosine kinase inhibitors that have been approved for the treatment of hepatocellular carcinoma (HCC). However, the different effects on tumor vasculature between lenvatinib and sorafenib are not well understood. In this study, we analyzed the effects of both drugs on vascular structure and function, including vascular normalization, and investigated whether the normalization had a positive effect on a combination therapy with the drugs and radiation using micro X-ray computed tomography with gold nanoparticles as a contrast agent, as well as immunohistochemical analysis and interstitial fluid pressure (IFP) measurement. In mice subcutaneously transplanted with mouse HCC cells, treatment with lenvatinib or sorafenib for 14 days inhibited tumor growth and reduced the tumor vessel volume density. However, analysis of integrated data on vessel density, rates of pericyte-covering and perfused vessels, tumor hypoxia, and IFP measured 4 days after drug treatment showed that treatment with 3 mg/kg of lenvatinib significantly reduced the microvessel density and normalized tumor vessels compared to treatment with 50 mg/kg of sorafenib. These results showed that lenvatinib induced vascular normalization and improved the intratumoral microenvironment in HCC tumors earlier and more effectively than sorafenib. Moreover, such changes increased the radiosensitivity of tumors and enhanced the effect of lenvatinib and radiation combination therapy, suggesting that this combination therapy is a powerful potential application against HCC.


Indomethacin prophylaxis, patent ductus arteriosus, and the risk of bronchopulmonary dysplasia: further analyses from the Trial of Indomethacin Prophylaxis in Preterms (TIPP).
To determine the risk of bronchopulmonary dysplasia (BPD) in subgroups of infants with and without patent ductus arteriosus (PDA) who were randomized to indomethacin prophylaxis or placebo, and to examine whether adverse drug effects on edema formation and oxygenation may explain why indomethacin prophylaxis does not reduce BPD. We studied 999 extremely low birth weight infants who participated in the Trial of Indomethacin Prophylaxis in Preterms (TIPP) and who survived to a postmenstrual age of 36 weeks. The incidence of BPD in the 2 subgroups of infants with PDA was 52% (55/105) after indomethacin prophylaxis and 56% (137/246) after placebo. In contrast, rates of BPD in the 2 subgroups without a PDA were 43% (170/391) after indomethacin prophylaxis and 30% (78/257) after placebo (P [interaction] = .015). Logistic regression analysis with adjustment for prognostic baseline factors showed that adverse and independent effects of indomethacin prophylaxis on the need for supplemental oxygen and on weight loss by the end of the first week of life may increase the risk of BPD in infants without PDA. Harmful side effects on oxygenation and edema formation may explain why indomethacin prophylaxis does not prevent BPD even though it reduces PDA.


Association of alcohol craving with alpha-synuclein (SNCA).
Studies have found that genomic variation in the gene SNCA, which encodes the protein alpha-synuclein, may contribute to the variation in alcohol consumption in an inbred rat model of alcohol preference. Studies in humans have provided support for an association between SNCA and craving for alcohol. To examine the role of this gene in alcohol dependence and related phenotypes, 30 single nucleotide polymorphisms (SNPs) were genotyped across the SNCA gene in a sample of 219 multiplex alcoholic families of European American descent. Two phenotypes, alcohol dependence and alcohol craving, were analyzed using the pedigree disequilibrium test. There was no evidence of association between any of the SNCA SNPs and alcohol dependence (p>or=0.13). In contrast, 8 SNPs provided evidence of association (p<0.05) with the phenotype of alcohol craving. Haplotype analysis further supported evidence of an association with alcohol craving; a haplotype encompassing SNPs in intron 4 through the region downstream of the gene was overtransmitted to cravers and a second haplotype was overtransmitted to noncravers. These results suggest that variation in SNCA contributes to alcohol craving, a common, although not uniform, feature of alcohol dependence.


Immune responses to CCAR1 and other dermatomyositis autoantigens are associated with attenuated cancer emergence.
BACKGROUNDThe temporal clustering of a cancer diagnosis with dermatomyositis (DM) onset is strikingly associated with autoantibodies against transcriptional intermediary factor 1-γ (TIF1-γ). Nevertheless, many patients with anti-TIF1-γ antibodies never develop cancer. We investigated whether additional autoantibodies are found in anti-TIF1-γ-positive patients without cancer.METHODSUsing a proteomic approach, we defined 10 previously undescribed autoantibody specificities in 5 index anti-TIF1-γ-positive DM patients without cancer. These were subsequently examined in discovery (n = 110) and validation (n = 142) cohorts of DM patients with anti-TIF1-γ autoantibodies.RESULTSWe identified 10 potentially novel autoantibodies in anti-TIF1-γ-positive DM patients, 6 with frequencies ranging from 3% to 32% in 2 independent DM cohorts. Autoantibodies recognizing cell division cycle and apoptosis regulator protein 1 (CCAR1) were the most frequent, and were significantly negatively associated with contemporaneous cancer (discovery cohort OR 0.27 [95% CI 0.7-1.00], P = 0.050; validation cohort OR 0.13 [95% CI 0.03-0.59], P = 0.008). When cancer did emerge, it occurred significantly later in anti-CCAR1-positive compared with anti-CCAR1-negative patients (median time from DM onset 4.3 vs. 0.85 years, respectively; P = 0.006). Cancers that emerged were more likely to be localized (89% of anti-CCAR1-positive cancers presenting at stage 0 or 1 compared with 42% of patients without anti-CCAR1 antibodies, P = 0.02). As the number of additional autoantibody specificities increased in anti-TIF1-γ-positive DM patients, the frequency of cancer decreased (P < 0.001).CONCLUSIONAs the diversity of immune responses in anti-TIF1-γ DM patients increases, the likelihood of cancer emerging decreases. Our findings have important relevance for cancer risk stratification in DM patients and for understanding natural immune regulation of cancer in humans.TRIAL REGISTRATIONNot applicable.FUNDING SOURCESThe NIH, the Donald B. and Dorothy L. Stabler Foundation, and the Huayi and Siuling Zhang Discovery Fund.


Generation of RCAS vectors useful for functional genomic analyses.
Avian leukosis type A virus-derived retroviral vectors have been used to introduce genes into cells expressing the corresponding avian receptor tv-a. This includes the use of Replication-Competent Avian sarcoma-leukosis virus (ASLV) long terminal repeat (LTR) with Splice acceptor (RCAS) vectors in the analysis of avian development, human and murine cell cultures, murine cell lineage studies and cancer biology. Previously, cloning of genes into this virus was difficult due to the large size of the vector and sparse cloning sites. To overcome some of the disadvantages of traditional cloning using the RCASBP-Y vector, we have modified the RCASBP-Y to incorporate "Gateway" site-specific recombination cloning of genes into the construct, either with or without HA epitope tags. We have found the repetitive "att" sequences, which are the targets for site-specific recombination, do not impair the production of infectious viral particles or the expression of the gene of interest. This is the first instance of site-specific recombination being used to generate retroviral gene constructs. These viral constructs will allow for the efficient transfer and expression of cDNAs needed for functional genomic analyses.


Triggering of T-lymphocytes via either T3-Ti or T11 surface structures opens a voltage-insensitive plasma membrane calcium-permeable channel: requirement for interleukin-2 gene function.
Stimulation of human T-lymphocytes via either the surface T3-Ti antigen-major histocompatibility complex receptor complex or the T11 molecule results in clonal proliferation through a calcium-dependent mechanism. To investigate this signal transduction, plasma membrane calcium-permeable channels were characterized in T-lymphocytes by means of whole cell or single channel patch-clamp recordings. Stimulation of T-lymphocytes via either structure results in opening of an identical set of voltage-insensitive plasma membrane Ca2+-permeable channels through the action of a diffusible second messenger. Previous work with excised inside-out patches suggests that inositol 1,4,5-trisphosphate is the activating second messenger of the voltage-insensitive T-cell Ca2+-permeable channel. Since there is a significant increase in phosphoinositide turnover after stimulation via either the T3-Ti or T11 pathway, it is suggested that triggering of either structure opens a common set of channels through this mechanism. Furthermore, currents flowing through Ca2+-permeable channels are apparently autoregulated, as inward conductance is abolished by elevation of Ca2+ concentration in the bathing solution. In particular, the steady-state rise in interleukin-2 (T-cell growth factor) mRNA is dependent on the rise of [Ca2+]i resulting from ion movement across this channel.


Identification of a sequence within the mouse metallothionein-I gene promoter mediating its basal transcription and of a protein interacting with this element.
Previous studies in our laboratory have shown that a trans-acting factor, which binds to a 106 bp sequence in the mouse metallothionein-I (MT-I) gene, is responsible for the relatively high level of MT-I gene transcription in the liver. Using electrophoretic mobility shift assay, we have now identified a 26 bp sequence within the 106 bp region, which interacts with a trans-activating factor in the liver nuclear extract. This sequence, designated MRE-c', is located between positions -135 and -110 with respect to the transcription start site and comprises the metal regulatory element MRE-c and part of its 5' and 3' flanking sequences. UV cross-linking and Southwestern analysis showed that a protein of an apparent molecular mass of 33,000 specifically interacts with MRE-c'. Deletion of the MRE-c' region resulted in a six- to sevenfold decrease in the MT-I promoter activity, as measured by reduction in chloramphenicol acetyltransferase activity. A comparison of other regulatory domains of the MT-I gene and the potential factors interacting with these sequences indicates that MRE-c' and probably the 33 kDa polypeptide are involved in the constitutive transcription of the MT-I gene.


Emergency ultrasound and urinalysis in the evaluation of flank pain.
To determine the sensitivity and specificity of limited emergency ultrasonography of the kidney in diagnosing renal colic. This was a prospective observational trial from December 2001 to December 2003 at a suburban emergency department. Patients who presented with flank pain suspicious for renal colic were enrolled. Exclusion criteria included fever, trauma, known current kidney stone, unstable vital signs, and inability to provide consent. All patients underwent sequential emergency ultrasonography and computed tomography of the kidneys and bladder. Data were analyzed using chi-square analysis. The primary outcome was the sensitivity and specificity of ultrasonography. Results were also stratified for presence of hematuria. Fifty-eight of the 104 patients enrolled in the study were diagnosed with renal colic. The overall sensitivity and specificity of bedside ultrasonography for the detection of hydronephrosis were 86.8 (95% confidence interval [CI] = 78.8 to 92.3) and 82.4 (95% CI = 74.1 to 88.1), respectively. In patients with hematuria, hydronephrosis by emergency ultrasonography demonstrated a sensitivity and specificity of 87.8 (95% CI = 80.3 to 92.5) and 84.8 (95% CI = 73.7 to 91.9), respectively. In 55 of the cases, the initial computed tomograph was read by a resident and later re-read by an attending physician. Using the reading of the attending physician as the criterion standard resulted in a sensitivity and specificity of 83.3 (95% CI = 73.2 to 88.0) and 92.0 (95% CI = 79.9 to 97.6), respectively. Emergency ultrasonography of the kidneys shows very good sensitivity and specificity for diagnosing renal colic in patients with flank pain and hematuria.


The synthesis of cytosolic ascorbate peroxidases in germinating seeds and seedlings of soybean and their behavior under flooding stress.
Cytosolic ascorbate peroxidases (cAPXs) of soybean have been found by proteome analysis to be downregulated in submerged seedlings. To elucidate the physiological meaning of this downregulation, soybean cAPXs were characterized in this study. Vigorous synthesis was detected in germinating seeds and seedlings. Expression of the corresponding genes was detected clearly in tissues that actively underwent cell division. The gene expression was suppressed by flooding stress, but not by salinity, cold or drought stress. The expression recovered 1 d after release from flooding stress, accompanied by growth resurgence.


Two-state conformational equilibrium in the Par-4 leucine zipper domain.
Prostate apoptosis response factor-4 (Par-4) is a pro-apoptotic and tumor-suppressive protein. A highly conserved heptad repeat sequence at the Par-4 C-terminus suggests the presence of a leucine zipper (LZ). This C-terminal region is essential for Par-4 self-association and interaction with various effector proteins. We have used nuclear magnetic resonance (NMR) spectroscopy to fully assign the chemical shift resonances of a peptide comprising the LZ domain of Par-4 at neutral pH. Further, we have investigated the properties of the Par-4 LZ domain and two point mutants under a variety of conditions using NMR, circular dichroism (CD), light scattering, and bioinformatics. Results indicate an environment-dependent conformational equilibrium between a partially ordered monomer (POM) and a predominantly coiled coil dimer (CCD). The combination of techniques used allows the time scales of the equilibrium to be probed and also helps to identify features of the amino acid sequence that may influence the equilibrium.


Effect of vitamin D, calcium and multiple micronutrients supplementation on lipid profile in pre-menopausal Bangladeshi garment factory workers with hypovitaminosis D.
Elevated total cholesterol and low-density lipoprotein cholesterol in sera are both well-known risk factors of coronary heart disease. Adequate vitamin D status is important for optimal function of many organs and tissues of our body. There is continuing controversy about the effect of adequate vitamin D consumption on serum lipids and lipoproteins. The present study assessed the effect of vitamin D, calcium and multiple micronutrients supplementation on the lipid profile in Bangladeshi young female garment factory workers who have hypovitaminosis D. This placebo-controlled intervention trial conducted over a period of one year randomly assigned a total of 200 apparently healthy subjects aged 16-36 years to 4 groups. The subjects received daily supplements of 400 IU of vitamin D (VD group) or 400 IU of vitamin D+600 mg of calcium lactate (VD-Ca group), or multiple micronutrients with 400 IU of vitamin-D+600 mg of calcium lactate (MMN-VD-Ca group), or the group consuming placebo (PL group). Serum concentrations of lipid and lipoprotein, 25-hydroxyvitamin D (25OHD) and intact parathyroid hormone (iPTH) were measured at baseline and after one year of follow-up. No significant changes in the serum levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), LDL-C/HDL-C ratio were observed in the supplemented groups compared to the placebo group. Supplementation had a positive effect (p<0.05) on very low-density-lipoprotein cholesterol (VLDL-C) and triacylglycerol (TAG). A negative correlation between changes in serum iPTH and HDL-C was observed, which indicated that subjects with the greatest decline in S-iPTH had the greatest increase in HDL-C. The results suggest that consumption of adequate vitamin D with calcium or MMN for one-year may have no impact on serum lipid profile in the subjects studied. Longer-term clinical trials with different doses of supplemental vitamin D are warranted in evaluating the effect of intervention.


Lack of association between Leber's hereditary optic neuropathy primary point mutations and multiple sclerosis in Iran.
The hypothesis that mitochondrial genes may implicate susceptibility to multiple sclerosis (MS) is supported by an increasing number of case reports on Leber's hereditary optic neuropathy (LHON)-associated mitochondrial DNA (mtDNA) point mutations in patients with MS. A number of mtDNA mutations with primary pathogenic significance for LHON, a maternally inherited disease causing severe bilateral visual loss predominantly in young men, have been detected in patients with an MS-like phenotype. To evaluate the link between MS and LHON primary point mutations, we investigated 31 non-related Iranian clinically definite MS patients (23 females and 8 males) with optic nerve involvement, as well as 25 patients (16 females and 9 males) without involvement of the optic nerve as controls. Three patients had severe bilateral visual loss without any recovery. We searched for the presence of LHON mitochondrial mutations at nucleotide positions (np) 11,778, 3,460, and 14,484 by mutation-specific polymerase chain reaction and restriction fragment length polymorphism. Our results suggest that there is no association between Iranian patients with MS and mtDNA point mutations at np 11,778, 3,460, and 14,484.


Low density lipoproteins reconstituted with steroids containing the nitrobenzoxadiazole fluorophore.
A new cholesterol analog, N-(7-nitrobenz-2-oxa-1,3-diazole)-23,24-dinor-5-cholen-22-amine-3 beta-ol, with fluorescent properties similar to those of fluorescein, has been synthesized. The fluorescence lifetimes, quantum yields, and wavelength maxima of N-(7-nitrobenz-2-oxa-1,3-diazole)-23,24-dinor-5-cholen-22-amine-3 beta-ol and its linoleate ester were solvent-dependent. The cholesterol analog was a satisfactory substrate for lecithin:cholesterol acyltransferase. The fluorescent sterol, added in ethanol, gave half-maximal suppression of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in cultured human fibroblasts at 2.5 microM and was twice as effective as cholesterol. The fluorescent steryl ester, incorporated into low density lipoprotein, was used to demonstrate high affinity cellular uptake and degradation of the reconstituted lipoproteins, intracellular accumulation of the free sterol and simultaneous suppression of 3-hydroxy-3-methylglutaryl coenzyme a reductase in fibroblasts. Half-maximal suppression was achieved at 10 micrograms mL-1 of low density lipoproteins reconstituted with the fluorescent steryl ester, compared to the same degree of suppression produced by 2 micrograms mL-1 of native low density lipoproteins. The interaction of low density lipoproteins reconstituted with N-(7-nitrobenz-2-oxa-1,3-diazole)-23,24-dinor-5-cholen-22-amine-3 beta-ol linoleate with cells was readily visualized by fluorescence microscopy and quantified by fluorimetry. These analogs will facilitate the studies of lipoprotein-cell interactions and phospholipid vesicle-cell interactions, the selection of cell mutants defective in lipoprotein metabolism, and the assessment of the immediate environment of the steroids in cellular membranes.


Improving Information Sharing for Youth in Foster Care.
There are ∼443 000 children in child protective custody (ie, foster care) in the United States. Children in protective custody have more medical, behavioral, and developmental problems that require health care services than the general population. These health problems are compounded by poor information exchange impeding care coordination. Health care providers often do not know which of their patients are in protective custody and are not privy to the critical social history collected by child protective services, including placement history and maltreatment history. Meanwhile, the custodial child protection agency and designated caregivers (ie, foster caregivers and kinship providers) often lack vital elements of the health history of children in their care, which can result in poor health care delivery such as medication lapses, immunization delay, and poor chronic disease management. In this case study, we address this critical component of health care delivery for a vulnerable population by describing a process of developing an information sharing system between health care and child welfare organizations in collaboration with child protection community partners. Lessons learned include recommended steps for improved information sharing: (1) develop shared community vision, (2) determine shareable information components, (3) implement and analyze information sharing approaches, and (4) evaluate information sharing efforts. A successful example of advocating for improvement of information sharing for youth in protective custody is explored to highlight these steps. In collaboration with child protective services, pediatricians can improve information sharing to impact both health care delivery and child protection outcomes.


Monodisperse titania microspheres via controlled nanoparticle aggregation.
This paper describes the fabrication of highly monodisperse TiO(2) nanoparticle aggregates (NPAs) by controlled aggregation of nanoparticles in a water-in-oil emulsion. Equally sized drops containing a titanium dioxide nanoparticle suspension are produced in a T-channel device. This procedure has a high tuning potential. Increasing the velocity of the titania suspension phase leads to an enlargement of the droplets, while raising the velocity of the oil phase reduces the drop diameter. The technique enables the preparation of monodisperse (<2%) drops between 150 and 400 μm in diameter. Evaporation of the suspension medium leads to nanoparticle aggregates. There is no significant change in dispersity from emulsion drops to NPAs, if the drying is carried out slowly.


[Behavior of the other eye in measuring fixation disparity].
In the subjective measurement of FD, the proband fuses contours presented in the peripheral macular areas of both eyes ('fusion lock'). The position of both eyes relative to each other is monitored by means of two haploscopically seen vertical lines presented in the central visual field, one above and one below a binocularly seen horizontal line. The subject is instructed to shift one of the vertical lines horizontally until the two are aligned, while fixating their intersection with the horizontal line. It has recently been questioned whether the foveolae really are pointed at the perceived intersection. To investigate this matter, we registered the position of one eye while intermittently covering the fellow eye, while the subject maintained fixation of the intersection. We found slight differences in monocular eye position depending on the presence or absence of fusion in the macular periphery, confirming earlier findings in part. These differences were more pronounced in the nondominant eye.


[Peripheral neuropathy and idiopathic hypereosinophilic syndrome: review of the literature apropos of a case with atypical presentation].
Involvement of the nervous system in the idiopathic hypereosinophilic syndrome is a common finding, peripheral neuropathy being the most frequent neurologic manifestation, usually appearing at the onset or following the appearance of eosinophilia. We describe here the case of a 42 year-old male patient with persistent eosinophilia (greater than 1500/mm3) for a period greater than six months in the absence of a known cause for the eosinophilia with end-organ injury, in whom the neurologic involvement preceded by months the elevation of peripheral blood eosinophil count, and consisted of peripheral neuropathy and the unusual involvement of a cranial nerve. There was severe distal wasting and weakness of the four limbs with sensory abnormalities which developed over a period of four months, along with left-sided deafness which was due to eighth cranial nerve involvement. Electrodiagnostic studies showed generalized nerve involvement of the axonal type. Nerve biopsy did not reveal eosinophilic infiltration or vasculitic changes, suggesting that neurotoxic eosinophil products may play a role in the neuropathy. There was marked improvement of the eosinophilia and slight improvement of the sensorimotor disturbance in the course of corticosteroid therapy.


ALK-activating homologous mutations in LTK induce cellular transformation.
Leukocyte tyrosine kinase (LTK) is a receptor tyrosine kinase reported to be overexpressed in human leukemia. Though much regarding the function of LTK remains unknown, it shares a high degree of similarity with anaplastic lymphoma kinase (ALK), which is found mutated in human cancer. In order to determine if LTK has transforming potential, we created two LTK mutants, F568L and R669Q, that correspond to two well-characterized activating mutations of ALK (F1174L and R1275Q). LTK-F568L, but not wildtype LTK or LTK-R669Q, transformed hematopoietic cells to cytokine independence. LTK-F568L exhibited a stronger ability to induce loss of contact inhibition and anchorage-independent growth of epithelial cells compared to LTK-R669Q, while wildtype LTK was non-transforming in the same cells. Likewise, LTK-F568L induced greater neurite outgrowth of PC12 cells than R669Q, while wildtype LTK could not. Correlating with transforming activity, LTK-F568L displayed significantly enhanced tyrosine phosphorylation compared to wildtype LTK and LTK-R668Q and induced activation of various signaling proteins including Shc, ERK and the JAK/STAT pathway. Expression of wildtype LTK or LTK-R669Q generally led to weaker activation of signaling proteins than expression of LTK-F568L, or no activation at all. Thus, mutating LTK at residue F568, and to a lesser extent at R669, activates the receptor tyrosine kinase, inducing cell signaling that results in transforming properties. These studies suggest that aberrant activation of LTK may contribute to neoplastic cell growth.


Hepatitis B virus reactivation in patients with resolved hepatitis B virus infection receiving chemotherapy or immunosuppressive therapy.
The role of antiviral prophylaxis before chemotherapy or immunosuppressive therapy to prevent hepatitis B virus (HBV) reactivation in patients with resolved HBV infection [hepatitis B surface antigen (HBsAg) negative, hepatitis B core antibody (anti-HBc) positive] is unclear. This study aimed to evaluate the efficacy of prophylactic antiviral therapy and outcomes of HBsAg-negative, anti-HBc-positive patients who received chemotherapy or immunosuppressive therapy. We retrospectively evaluated the medical records of HBsAg-negative, anti-HBc-positive patients who underwent chemotherapy or immunosuppressive therapy from January 2013 through November 2016 at a single institute in southern Taiwan. Among 1000 included HBsAg-negative, anti-HBc-positive patients, the rate of hepatitis B surface antibody (anti-HBs) seropositivity before chemotherapy or immunosuppressive therapy was 76.6%. Twenty-six patients received a prophylactic oral antiviral agent (one telbuvudine, two lamivudine, 22 entecavir, and one tenofovir). Seven (0.7%) patients were diagnosed with HBV reactivation during or after chemotherapy courses. In multivariate Cox regression analysis, an rituximab-based regimen (hazard ratio: 11.74; 95% confidence interval: 1.62-84.94; P=0.02) and baseline anti-HBs-positive status (hazard ratio: 0.17; 95% confidence interval: 0.04-0.8; P=0.03) were significant predictive factors for HBV reactivation. Among anti-HBs-negative recipients of rituximab-based chemotherapy, HBV reactivation was observed in zero of nine patients who received prophylactic antiviral therapy and three (33.3%) of nine patients who did not. Negative anti-HBs status and rituximab-containing regimens are both important factors for predicting chemotherapy or immunosuppressive therapy-related HBV reactivation in patients with resolved HBV infection. Therefore, antiviral prophylaxis should be considered in this patient population.


Seroprevalence of Brucella spp. and Rift Valley fever virus among slaughterhouse workers in Isiolo County, northern Kenya.
Brucella spp. and Rift Valley fever virus (RVFV) are classified as priority zoonotic agents in Kenya, based on their public health and socioeconomic impact on the country. Data on the pathogen-specific and co-exposure levels is scarce due to limited active surveillance. This study investigated seroprevalence and co-exposure of Brucella spp. and RVFV and associated risk factors among slaughterhouse workers in Isiolo County, northern Kenya. A cross-sectional serosurvey was done in all 19 slaughterhouses in Isiolo County, enrolling 378 participants into the study. The overall seroprevalences for Brucella spp. and RVFV were 40.2% (95% CI: 35.2-45.4) and 18.3% (95% CI: 14.5-22.5), respectively while 10.3% (95% CI 7.4%-13.8%) of individuals were positive for antibodies against both Brucella spp. and RVFV. Virus neutralisation tests (VNT) confirmed anti-RVFV antibodies in 85% of ELISA-positive samples. Our seroprevalence results were comparable to community-level seroprevalences previously reported in the area. Since most of the study participants were not from livestock-keeping households, our findings attribute most of the detected infections to occupational exposure. The high exposure levels indicate slaughterhouse workers are the most at-risk population and there is need for infection, prevention, and control programs among this high-risk group. This is the first VNT confirmation of virus-neutralising antibodies among slaughterhouse workers in Isiolo County and corroborates reports of the area being a high-risk RVFV area as occasioned by previously reported outbreaks. This necessitates sensitization campaigns to enhance awareness of the risks involved and appropriate mitigation measures.


[First results from the OBSERVANT II study: clinical characteristics of patients with aortic stenosis undergoing transcatheter treatment with new generation devices].
The OBSERVANT study evaluated the medium and long-term outcomes in patients undergoing transcatheter aortic valve implantation (TAVI) versus surgical aortic valve replacement obtaining comparative effectiveness results at least for the first-generation devices. The OBSERVANT II study enrolled a new series of TAVI procedures to evaluate whether the use of new-generation devices has modified outcome differences recorded in the OBSERVANT study. The aim of this paper is to describe the clinical characteristics of the OBSERVANT II population and compare them with those of the OBSERVANT population. OBSERVANT II is a prospective multicenter observational study, which enrolled patients with severe aortic stenosis, who underwent a TAVI procedure in 30 Italian centers from December 2016 to September 2018. Clinical, anatomical and procedural information was recorded for each patient as well as procedure characteristics and any periprocedural outcomes. An administrative follow-up will allow to obtain information on medium and long-term outcomes. The enrolled population consists of 3001 patients with a mean age similar to that recorded in OBSERVANT (81.7 ± 6.3 vs 81.9 ± 6.2 years; p=0.336). OBSERVANT II patients show a lower prevalence of several morbid conditions than OBSERVANT patients (i.e. chronic obstructive pulmonary disease, coronary artery disease, peripheral arterial disease, etc.), though having very similar mean values of EuroSCORE II (7.2 ± 7.6% vs 7.4 ± 7.9%; p=0.558). According to three EuroSCORE II risk classes (≤3; 3-7; ≥7), the in-hospital mortality rates for patients enrolled in the two studies are 1.0% vs 2.0% (p≤0.000), 1.4% vs 4.1% (p<0.000) and 2.4% vs 7.7% (p<0.000), respectively. The first results of the OBSERVANT II study show that patients undergoing TAVI with new-generation devices are elderly, have a lower risk profile than patients enrolled in the OBSERVANT study but comparable mean values of EuroSCORE II. The in-hospital mortality rate for patients enrolled in the OBSERVANT II study is lower than that recorded in the OBSERVANT study across all the EuroSCORE II risk classes.


Identification of specific proteins synthesized by type II pneumocytes in primary culture.
Proteins from primary cultures of type II granular pneumocytes have been examined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis to identify type II cell-specific proteins. The distribution of Coomassie Blue-stained bands in preparations of cellular proteins, culture medium, lavage and lamellar bodies have been compared. The most prominent stained band in the serum-free medium from type II cell cultures (HS1; Mr 39900) corresponds to a major protein in acellular sedimentable (20000 g for 30 min) crude surfactant obtained from rat lungs by saline (0.9% NaCl) lavage. A second protein (HS2; Mr 12000) is also found both in type II cell-conditioned medium and in lavage. Neither rat serum nor donor calf serum (used in the isolation of the type II cells) contains a protein co-migrating with HS1 or HS2 proteins. HS1 is also found in Coomassie Blue-stained gels of cellular proteins and of lamellar bodies isolated from whole lungs. Cultures of type II cells incorporate [14C]phenylalanine into HS1 and HS2 as shown by autoradiography of sodium dodecyl sulphate/polyacrylamide gels of culture medium. Rat lungs perfused in situ incorporate [35S]methionine into HS1 in the lamellar body fraction. A third protein (HS3; Mr 47000) is observed only in autoradiographs of cell culture medium; no corresponding Coomassie Blue-stained band can be identified in medium, in cells or in lung lavage. No protein bands corresponding to HS1, HS2 or HS3 are found in conditioned media from pulmonary alveolar macrophages, rat fibroblasts or bovine aorta endothelial cells. Two-dimensional gel electrophoresis of HS1 shows a single polypeptide with an isoelectric point of 6.3; HS3 appears as a chain of spots with a range of isoelectric points from 6.3 to 6.6. HS2 has not been identified on two-dimensional gels. The amino acid composition of HS1 does not differ significantly from that of surfactant apoproteins studied previously; however, HS1 is not detected by glycoprotein stains, nor does it appear to be a subunit of a thiol-linked multimer.


Minimizing Oxidation of Freeze-Dried Monoclonal Antibodies in Polymeric Vials Using a Smart Packaging Approach.
Primary containers made of cyclic olefin polymer (COP) have recently gained attention since they may overcome several risks and shortcomings of glass containers as they exhibit a high break resistance, biocompatibility, and homogeneous heat transfer during lyophilization. On the downside, COP is more permeable for gases, which can lead to an ingress of oxygen into the container over time. Since oxidation is an important degradation pathway for monoclonal antibodies (mAbs), the continuous migration of oxygen into drug product containers should be avoided overall. To date, no long-term stability studies regarding lyophilizates in polymer vials have been published, potentially because of the unbearable gas permeability. In this study, we demonstrate that after lyophilization in COP vials and storage of these vials in aluminum pouches together with combined oxygen and moisture absorbers ("smart packaging"), oxidation of two lyophilized therapeutic antibodies was as low as in glass vials due to the deoxygenated environment in the pouch. Nevertheless, active removal of oxygen from the primary container below the initial level over time during storage in such "smart" secondary packaging was not achieved. Furthermore, residual moisture was controlled. Overall, the smart packaging reveals a promising approach for long-term stability of biopharmaceuticals; in addition to COP's known benefits, stable, low oxygen and moisture levels as well as the protection from light and cushioning against mechanical shock by the secondary packaging preserve the sensitive products very well.


Rescue of cardiomyocyte dysfunction by phospholamban ablation does not prevent ventricular failure in genetic hypertrophy.
Cardiac hypertrophy, either compensated or decompensated, is associated with cardiomyocyte contractile dysfunction from depressed sarcoplasmic reticulum (SR) Ca(2+) cycling. Normalization of Ca(2+) cycling by ablation or inhibition of the SR inhibitor phospholamban (PLN) has prevented cardiac failure in experimental dilated cardiomyopathy and is a promising therapeutic approach for human heart failure. However, the potential benefits of restoring SR function on primary cardiac hypertrophy, a common antecedent of human heart failure, are unknown. We therefore tested the efficacy of PLN ablation to correct hypertrophy and contractile dysfunction in two well-characterized and highly relevant genetic mouse models of hypertrophy and cardiac failure, Galphaq overexpression and human familial hypertrophic cardiomyopathy mutant myosin binding protein C (MyBP-C(MUT)) expression. In both models, PLN ablation normalized the characteristically prolonged cardiomyocyte Ca(2+) transients and enhanced unloaded fractional shortening with no change in SR Ca(2+) pump content. However, there was no parallel improvement in in vivo cardiac function or hypertrophy in either model. Likewise, the activation of JNK and calcineurin associated with Galphaq overexpression was not affected. Thus, PLN ablation normalized contractility in isolated myocytes, but failed to rescue the cardiomyopathic phenotype elicited by activation of the Galphaq pathway or MyBP-C mutations.


Seeing voices: High-density electrical mapping and source-analysis of the multisensory mismatch negativity evoked during the McGurk illusion.
Seeing a speaker's facial articulatory gestures powerfully affects speech perception, helping us overcome noisy acoustical environments. One particularly dramatic illustration of visual influences on speech perception is the "McGurk illusion", where dubbing an auditory phoneme onto video of an incongruent articulatory movement can often lead to illusory auditory percepts. This illusion is so strong that even in the absence of any real change in auditory stimulation, it activates the automatic auditory change-detection system, as indexed by the mismatch negativity (MMN) component of the auditory event-related potential (ERP). We investigated the putative left hemispheric dominance of McGurk-MMN using high-density ERPs in an oddball paradigm. Topographic mapping of the initial McGurk-MMN response showed a highly lateralized left hemisphere distribution, beginning at 175 ms. Subsequently, scalp activity was also observed over bilateral fronto-central scalp with a maximal amplitude at approximately 290 ms, suggesting later recruitment of right temporal cortices. Strong left hemisphere dominance was again observed during the last phase of the McGurk-MMN waveform (350-400 ms). Source analysis indicated bilateral sources in the temporal lobe just posterior to primary auditory cortex. While a single source in the right superior temporal gyrus (STG) accounted for the right hemisphere activity, two separate sources were required, one in the left transverse gyrus and the other in STG, to account for left hemisphere activity. These findings support the notion that visually driven multisensory illusory phonetic percepts produce an auditory-MMN cortical response and that left hemisphere temporal cortex plays a crucial role in this process.


Effects of quality on outcomes in primary care: a review of the literature.
It is widely believed that health care quality affects primary care outcomes, but the evidence is fragmented and incomplete. The authors searched MEDLINE for relevant articles published between 1950 and 2006 and reviewed the evidence to assess the relationship between the personal aspects of primary care quality and patients' health status and health services utilization. These personal aspects, which include patient-physician continuity and communication, are distinct from the technical aspects of primary care, which include ordering tests, treatments, and referrals. Fourteen articles met the inclusion criteria. Results showed that greater continuity of care is associated with less use of hospitals and emergency departments and lower health care costs; effective communication may be associated with better health status. The limited available evidence suggests that higher quality in the personal aspects of primary care is associated with some but not all outcomes of care. Additional research is needed to define these relationships more clearly.


Essential role for Smad3 in angiotensin II-induced tubular epithelial-mesenchymal transition.
Angiotensin II (Ang II) is a key mediator of chronic kidney disease, in which epithelial-mesenchymal transition (EMT) is a critical process mediated by the TGFbeta/Smad signalling pathway. The present study examined the specific role of Smads in Ang II-induced EMT in vitro and in vivo. We found that Ang II signalled through the receptor of AT1, not AT2, to activate Smad2/3 and induce EMT in a normal rat tubular epithelial cell line (NRK52E). Activation of Smads by Ang II was attributed to degradation of an inhibitory Smad7, which was mediated by the AT1-Smurf2-dependent ubiquitin degradation mechanism because blockade of AT1 receptor or knockdown of Smurf2 inhibited Smad7 loss, thereby reducing Smad2/3 activation and EMT in response to Ang II. In contrast, over-expression of Smad7 inhibited Ang II-induced Smad2/3 activation and EMT in NRK52E cells and in a rat model of remnant kidney disease. Moreover, knockdown of Smad3, not Smad2, attenuated Ang II-induced EMT. In conclusion, Ang II activates Smad signalling to induce EMT, which is mediated by a loss of Smad7 through the AT1-Smurf2-dependent ubiquitin degradation pathway. Smad3, but not Smad2, may be a mediator of EMT, while Smad7 may play a protective role in EMT in response to Ang II.


Effect of Postoperative Bleaching on Microleakage of Etch-and-Rinse and Self-etch Adhesives.
Bleaching the discoloured teeth may affect the tooth/composite interface. The aim of this in vitro experimental study was to evaluate the effect of vital tooth bleaching on microleakage of existent class V composite resin restorations bonded with three dental bonding agents. Class V cavities were prepared on buccal surfaces of 72 intact, extracted human anterior teeth with gingival margins in dentin and occlusal margins in enamel, and randomly divided into 3 groups. Cavities in the three groups were treated with Scotch bond Multi-Purpose, a total etch system and Prompt L-Pop and iBond, two self-etch adhesives. All teeth were restored with Z250 resin composite material and thermo-cycled. Each group was equally divided into the control and the bleached subgroups (n = 12). The bleached subgroups were bleached with 15% carbamide peroxide gel for 8 hours a day for 15 days. Microleakage scores were evaluated on the incisal and cervical walls. Data were analyzed using Kruskal-Wallis, Mann-Whitney and Bonferroni post-hoc tests (α = 0.05). Bleaching with carbamide peroxide gel significantly increased the microleakage of composite restorations in Prompt L-Pop group at dentinal walls (P = 0.001). Bleaching had no effect on microleakage of restorations in the Scotch bond Multi-Purpose and iBond groups. Vital tooth bleaching with carbamide peroxide gel has an adverse effect on marginal seal of dentinal walls of existent composite resin restorations bonded with prompt L-Pop self-etch adhesive.


Listeria monocytogenes sequence type 6 and increased rate of unfavorable outcome in meningitis: epidemiologic cohort study.
We analyzed clinical characteristics, treatment, genetic diversity, and outcome of 92 adults with Listeria monocytogenes meningitis included in 2 prospective nationwide cohort studies. Episodes of community-acquired listerial meningitis confirmed by cerebrospinal fluid culture were included from 1998 to 2002 and 2006 to 2012. We compared patients and pathogen characteristics between cohorts and identified predictors for an unfavorable outcome according to the Glasgow Outcome Scale. Thirty episodes were included from 1998 to 2002 and 62 from 2006 to 2012; clinical and laboratory characteristics on admission were similar between cohorts. However, the rate of unfavorable outcome increased from 27% in the 1998-2002 cohort to 61% in the 2006-2012 cohort (P = .002). Differences between cohorts were increased use of adjunctive dexamethasone therapy (0% in 1998-2002 vs 53% in 2006-2012; P < .001) and emergence of infection by L. monocytogenes genotype sequence type 6 (ST6; 4% in 1998-2002 vs 29% in 2006-2012; P = .009). Multivariate regression analysis identified infection with L. monocytogenes ST6 as the sole predictor of unfavorable outcome (odds ratio, 3.77; 95% confidence interval, 1.07-13.33). Patients infected with genotypes other than ST6 also had an increased rate of unfavorable outcome over time (P = .03). The rate of unfavorable outcome among adults with listerial meningitis has increased over a 14-year period, from 27% to 61%. The emerging L. monocytogenes genotype ST6 was identified as the main factor leading to poorer prognosis. Adjunctive dexamethasone may be discontinued if L. monocytogenes is identified, as there is no proven benefit in Listeria meningitis.


Shisha smoking: impact on cognitive functions impairments in healthy adults.
Shisha smoking is becoming highly prevalent and is a predominant mode of tobacco use particularly amongst the young generation around the globe. This study aims to investigate whether or not shisha smoking impairs the cognitive functioning in adolescents. We recruited 65 (32 control subjects and 33 shisha smokers) apparently healthy male volunteers. The first group consisted of 33 volunteer male shisha smoker subjects with mean age 24.45±2.93 (mean ± SD) years. The second group consists of 32 subjects with mean age 23.31±2.68 (mean ± SD) years who did not smoke shisha. Age, gender, weight, height, ethnicity, educational and socioeconomic status of a matched design was used to appraise the impact of shisha smoking on cognitive functions. The cognitive function's outcome variables were the response time (Attention Switching Task-AST), (Complex Reaction Time-CRT task) and the percentage of correct answers (Pattern Recognition Memory PRM-task). Cognitive functions were assessed by using Cambridge Neuropsychological Automated Battery (CANTAB). Shisha smokers showed significant decline in cognitive performances AST-Latency (p=0.03); AST-Congruent (p=0.05); AST-Incongruent (p=0.05); mean CRT (p=0.001); CRT % (p=0.01); PRM (p=0.024) compared to their matched control. Cognitive functions including attention, alertness, and memory were significantly impaired in healthy adult shisha smokers compared to non-shisha smokers.


Combination of single quantitative parameters into multiparametric model for ischemia detection is not superior to visual assessment during dobutamine stress echocardiography.
To evaluate if the combination of several quantitative parameters into a mathematical model would enhance the detection of myocardial ischemia during dobutamine stress echocardiography (DSE) when compared to conventional wall motion analysis. In a prospective study design 151 patients (age 61.8 ± 9.2) in test group and 105 patients (age 64.0 ± 10.6) in validation group were selected and underwent DSE between January 2008 and December 2012. In all patients coronary angiography was performed within 6-8 weeks from DSE, considering at least one stenosis ≥50% per patient as significant coronary artery disease (CAD). Results of DSE visual assessment and myocardial velocity, strain and strain rate parameters derived from speckle tracking imaging were imported automatically to an originally created software. A mathematical model calculating prognosis of at least one stenosis per patient and stenosis in separate arteries was constructed. Myocardial ischemia was visually detected in 60 (39.7 %) and in 58 (54.2 %) patients of the test and validation group, respectively. A total of 76 (50.3 %) patients in the test group and 69 patients (65.7 %) in the validation group had ≥50% coronary stenosis. Sensitivity and specificity of the mathematical model per patient in the test group were 91.6 % and 86.3 % compared to 76.8 % and 89.0 % of the visual assessment, respectively. However, in the validation group the sensitivity, specificity, positive predictive value and negative predictive value dropped down significantly becoming lower to visual assessment. Myocardial deformation imaging may potentially replace visual assessment with an automated predictive model for stress-induced ischemia detection. However, a multiparametric mathematical model based on quantitative deformation markers did not demonstrate incremental value to visual assessment of wall motion.


Scientific Opinion on the assessment of the control measures for category A diseases of Animal Health Law: Foot and Mouth Disease.
EFSA received a mandate from the European Commission to assess the effectiveness of some of the control measures against diseases included in the Category A list according to Regulation (EU) 2016/429 on transmissible animal diseases ('Animal Health Law'). This opinion belongs to a series of opinions where these control measures will be assessed, with this opinion covering the assessment of control measures for foot and mouth disease (FMD). In this opinion, EFSA and the AHAW Panel of experts review the effectiveness of: i) clinical and laboratory sampling procedures, ii) monitoring period and iii) the minimum radius of the protection and surveillance zones, and the minimum length of time the measures should be applied in these zones. The general methodology used for this series of opinions has been published elsewhere; nonetheless, the transmission kernels used for the assessment of the minimum radius of the protection zone of 3 km and of the surveillance zone of 10 km are shown. Several scenarios for which these control measures had to be assessed were designed and agreed prior to the start of the assessment. The monitoring period of 21 days was assessed as effective, and it was concluded that the protection and the surveillance zones comprise > 99% of the infections from an affected establishment if transmission occurred. Recommendations, provided for each of the scenarios assessed, aim to support the European Commission in the drafting of further pieces of legislation, as well as for plausible ad hoc requests in relation to FMD.


A facilitated approach to family case conferencing for people with advanced dementia living in nursing homes: perceptions of palliative care planning coordinators and other health professionals in the IDEAL study.
Palliative care for nursing home residents with advanced dementia is often sub-optimal due to poor communication and limited care planning. In a cluster randomized controlled trial, registered nurses (RNs) from 10 nursing homes were trained and funded to work as Palliative Care Planning Coordinators (PCPCs) to organize family case conferences and mentor staff. This qualitative sub-study aimed to explore PCPC and health professional perceptions of the benefits of facilitated case conferencing and identify factors influencing implementation. Semi-structured interviews were conducted with the RNs in the PCPC role, other members of nursing home staff, and physicians who participated in case conferences. Analysis was conducted by two researchers using a thematic framework approach. Interviews were conducted with 11 PCPCs, 18 other nurses, eight allied health workers, and three physicians. Perceived benefits of facilitated case conferencing included better communication between staff and families, greater multi-disciplinary involvement in case conferences and care planning, and improved staff attitudes and capabilities for dementia palliative care. Key factors influencing implementation included: staffing levels and time; support from management, staff and physicians; and positive family feedback. The facilitated approach explored in this study addressed known barriers to case conferencing. However, current business models in the sector make it difficult for case conferencing to receive the required levels of nursing qualification, training, and time. A collaborative nursing home culture and ongoing relationships with health professionals are also prerequisites for success. Further studies should document resident and family perceptions to harness consumer advocacy.


Recognition of local anesthetic maldistribution in axillary brachial plexus block guided by ultrasound and nerve stimulation.
Nerve stimulation may occur despite the presence of a fascial barrier between the needle tip and the nerve, which may prevent appropriate flow or distribution of local anesthetic solution. During an axillary nerve block, ultrasound (US) guidance was used to identify the median nerve. Insertion of a needle with US and nerve stimulator guidance resulted in the appearance of the needle tip in contact with the nerve. However, as local anesthetic injection was begun, it was clear that the injectate was accumulating superficial to the investing fascia of the neurovascular bundle. No injectate was seen below the fascia. With US guidance, the needle was repositioned at a greater depth. Repeat injection of local anesthetic clearly flowed around the nerve.


Zinc-binding and protein-protein interactions mediated by the polyomavirus large T antigen zinc finger.
Polyomavirus large tumor antigen (LT) contains a potential C2H2 zinc binding element between residues 452 and 472. LT also contains a third histidine in this region, conserved among the polyomavirus LTs. Synthetic peptides of this region bound a single atom of zinc, as determined by spectroscopic analysis. Blotting experiments also showed that fusion proteins containing the element, as well as full-length LT, bound 65Zn. Polyomavirus middle T and small T antigens also bound zinc in the blotting assay. Site-directed mutagenesis showed the importance of this element in LT. Point mutations in four of the conserved residues (C-452, C-455, H-465, and H-469) blocked the ability of LT to function in viral DNA replication, while mutation of H-472-->L decreased replication to 1/30th that of the wild type. Point mutations in intervening residues tested had little effect on replication. Mutants resulting from mutations in the conserved cysteine or histidine residues retained the ability to bind origin DNA. However, they did show a defect in self-association. Because double-hexamer formation is involved in DNA replication, this deficiency is sufficient to explain the defect in replication. Mutants created by point mutations of the coordinating residues were also deficient in replication-associated phosphorylations.


Acute respiratory failure as a rare complication of celiac plexus block in a patient with adenocarcinoma of the ampulla of vater.
Celiac and splanchnic plexus blocks are considered as terminal approaches for pain control in end stage pancreatic cancer. It may be done temporarily (using local anesthetics) or as a permanent act (using alcohol and/or phenol). Like every other interventional procedure, celiac plexus block has its own potential complications and hazards among them pneumothorax and ARDS are very rare. In this case report we present an end stage patient with adenocarcinoma of ampulla of Vater with involvement of both abdomen and thorax who presented with severe intractable abdominal pain. Bilateral celiac plexus block in this patient resulted in left side pneumothorax and subsequent development of ARDS. We discuss the rare complications of celiac plexus block as well.


Gaming across different consoles: exploring the influence of control scheme on game-player enjoyment.
Many studies have investigated how different technological features impact the experience of playing video games, yet few have focused on how control schemes may affect the play experience. This research employed a between-subjects design to explore the relationship between the type of console played (Nintendo Wii, Playstation 2) and feelings of flow and enjoyment during the game-play experience. Results indicated that participants reported greater feelings of control and enjoyment with a traditional control scheme (Playstation 2) than with the more technologically advanced control scheme (Nintendo Wii). Further mediation analysis showed that enjoyment was driven by the sense of control that participants experienced and not simply by whether they won the game. Theoretical and practical implications are discussed.


Cloning and expression of a cloxacillin-hydrolyzing enzyme and a cephalosporinase from Aeromonas sobria AER 14M in Escherichia coli: requirement for an E. coli chromosomal mutation for efficient expression of the class D enzyme.
Two beta-lactamase genes, asbA1 and asbB1, encoding AsbA1 and AsbB1, respectively, have been cloned from Aeromonas sobria AER 14M into Escherichia coli. AsbA1 was expressed at low but detectable levels in all E. coli laboratory cloning strains tested. AsbB1 was expressed well in the E. coli cloning strain DH5 alpha. However, no enzyme activity could be detected from the same clone when placed in E. coli MC1061. Ampicillin-resistant mutants of E. coli MC1061 were obtained that expressed high levels of enzymatically active AsbB1. Four independent mutants were examined. All four mutations mapped to one locus, designated blpA (beta-lactamase permissive). The blpA locus was distinct from other known loci that play a role in beta-lactamase expression, i.e., the two loci that affect expression of the Bacteroides fragilis metallo-beta-lactamase and the ampC regulatory genes, ampD, ampE, and ampG. Sequence analysis of asbA1 and asbB1 revealed that AsbA1 was a class C beta-lactamase most closely related to the Pseudomonas aeruginosa chromosomal cephalosporinase and probably represents the common A. sobria cephalosporinase. AsbB1 was a class D enzyme most closely related to the oxacillin-hydrolyzing enzyme OXA-1, with 34% amino acid sequence identity. Purified AsbA1 was a typical cephalosporinase with a substrate profile that reflected high rates of hydrolysis of cephaloridine compared with benzylpenicillin. Purified AsbB1 showed strong penicillinase activity, with hydrolysis rates for carbenicillin and cloxacillin 2 to 2.5 times that for benzylpenicillin. Hydrolysis of imipenem was < or = 1% of that for benzylpenicillin. Both clavulanic acid and tazobactam strongly inhibited AsbB1, while sulbactam inhibited the AsbB1 enzyme less effectively. None of the inhibitors worked well against the AsbA1 enzyme. The chelators EDTA and 1,10-o-phenanthroline did not affect the activity of either enzyme. A. sobria AER 14M was found to produce both a group 1 cephalosporinase and a novel group 2d cloxacillin-hydrolyzing beta-lactamase that has been designated here OXA-12.


Unsupervised Instance Discriminative Learning for Depression Detection from Speech Signals.
Major Depressive Disorder (MDD) is a severe illness that affects millions of people, and it is critical to diagnose this disorder as early as possible. Detecting depression from voice signals can be of great help to physicians and can be done without any invasive procedure. Since relevant labelled data are scarce, we propose a modified Instance Discriminative Learning (IDL) method, an unsupervised pre-training technique, to extract augment-invariant and instance-spread-out embeddings. In terms of learning augment-invariant embeddings, various data augmentation methods for speech are investigated, and time-masking yields the best performance. To learn instance-spreadout embeddings, we explore methods for sampling instances for a training batch (distinct speaker-based and random sampling). It is found that the distinct speaker-based sampling provides better performance than the random one, and we hypothesize that this result is because relevant speaker information is preserved in the embedding. Additionally, we propose a novel sampling strategy, Pseudo Instance-based Sampling (PIS), based on clustering algorithms, to enhance spread-out characteristics of the embeddings. Experiments are conducted with DepAudioNet on DAIC-WOZ (English) and CONVERGE (Mandarin) datasets, and statistically significant improvements, with p-value 0.0015 and 0.05, respectively, are observed using PIS in the detection of MDD relative to the baseline without pre-training.


Iron overload in transfusion-dependent patients.
Before the advent of effective iron chelation, death from iron-induced cardiomyopathy occurred in the second decade in patients with transfusion-dependent chronic anemias. The advances in our understanding of iron metabolism; the ability to monitor iron loading in the liver, heart, pancreas and pituitary; and the availability of several effective iron chelators have dramatically improved survival and reduced morbidity from transfusion-related iron overload. Nevertheless, significantly increased survival brings about new complications such as malignant transformation resulting from prolonged exposure to iron, which need to be considered when developing long-term therapeutic strategies. This review discusses the current biology of iron homeostasis and its close relation to marrow activity in patients with transfusion-dependent anemias, and how biology informs clinical approach to treatment.


International medical graduates: the Medicentres experience.
The influx of foreign medical graduates into Canada presents both opportunity and challenges. The opportunity to increase the number of suitably and appropriately qualified providers to meet the demands of an aging and increasingly infirm population is clear. Some of the pressures on the health system could be met by integrating these graduates. However, identifying those with the necessary knowledge and skills to function at the same standard as Canadian graduates is challenging because the training and skills of many graduates is substandard requiring extensive rehabilitation. Furthermore there is a substantial deficiency of an evaluation/licensing pathway and funding for such an initiative.


Microbial battery for efficient energy recovery.
By harnessing the oxidative power of microorganisms, energy can be recovered from reservoirs of less-concentrated organic matter, such as marine sediment, wastewater, and waste biomass. Left unmanaged, these reservoirs can become eutrophic dead zones and sites of greenhouse gas generation. Here, we introduce a unique means of energy recovery from these reservoirs-a microbial battery (MB) consisting of an anode colonized by microorganisms and a reoxidizable solid-state cathode. The MB has a single-chamber configuration and does not contain ion-exchange membranes. Bench-scale MB prototypes were constructed from commercially available materials using glucose or domestic wastewater as electron donor and silver oxide as a coupled solid-state oxidant electrode. The MB achieved an efficiency of electrical energy conversion of 49% based on the combustion enthalpy of the organic matter consumed or 44% based on the organic matter added. Electrochemical reoxidation of the solid-state electrode decreased net efficiency to about 30%. This net efficiency of energy recovery (unoptimized) is comparable to methane fermentation with combined heat and power.


Leptospermum Honey for Wound Care in an Extremely Premature Infant.
Neonatal wound care is challenging due to the fragility and vulnerable skin structure. Neonates are often left susceptible to the forces of their environment, leaving them open to infection when skin injury occurs. Leptospermum honey has been used successfully in adult patients, with evidence lacking in the neonatal population. This case demonstrates the management of a difficult-to-heal wound in a 23-week gestation infant. Selecting the proper treatment and products for wound healing is challenging, with little evidence-based research available for the treatment of neonatal wounds. Leptospermum honey and other adult-driven dressings have been used for neonatal wound care as well as other adult-driven dressings. This case demonstrates the benefits of Leptospermum honey as an option for neonatal wounds. This case presents the treatment and healing of an extensive wound of a 23-week gestation neonate using a hydrogel product initially and then transitioning to a Leptospermum honey dressing due to suboptimal healing. Results of this treatment included quick healing time, little to no scarring, and no loss of movement or function to the affected extremities. The incorporation of Leptospermum honey for wound care has the potential to promote faster wound healing, with less scarring in the neonatal population. Adult wound care principles have been applied in the face of a weak evidence base relating to neonatal-specific cases. There is a need for continued research related to moist wound healing in the neonatal population, with resulting product and practice recommendations.


Purification, characterization and function of dihydrolipoamide dehydrogenase from the cyanobacterium Anabaena sp. strain P.C.C. 7119.
A dihydrolipoamide dehydrogenase (dihydrolipoamide: NAD+ oxidoreductase, EC 220.127.116.11) (DLD) has been found in the soluble fraction of cells of both unicellular (Synechococcus sp. strain P.C.C. 6301) and filamentous (Calothrix sp. strain P.C.C. 7601 and Anabaena sp. strain P.C.C. 7119) cyanobacteria. DLD from Anabaena sp. was purified 3000-fold to electrophoretic homogeneity. The purified enzyme exhibited a specific activity of 190 units/mg and was characterized as a dimeric FAD-containing protein with a native molecular mass of 104 kDa, a Stokes' radius of 4.28 nm and a very acidic pI value of about 3.7. As is the case with the same enzyme from other sources, cyanobacterial DLD showed specificity for NADH and lipoamide, or lipoic acid, as substrates. Nevertheless, the strong acidic character of the Anabaena DLD is a distinctive feature with respect to the same enzyme from other organisms. The presence of essential thiol groups was suggested by the inactivation produced by thiol-group-reactive reagents and heavy-metal ions, with lipoamide, but not NAD+, behaving as a protective agent. The function and physiological significance of Anabaena DLD are discussed in relation to the fact that 2-oxoacid dehydrogenase complexes have not been detected so far in filamentous cyanobacteria. Glycine decarboxylase activity, which might be involved in photorespiratory metabolism, has been found, however, in cell extracts of Anabaena sp. strain P.C.C. 7119 as the present study demonstrates.


Hormonal activation of type-L hormone-sensitive lipase measured in defatted heart powders.
Adrenaline, 3-isobutyl-1-methylxanthine (MIX) and dibutyryl cyclic AMP (Bt2 cyclic AMP) stimulated type-L hormone-sensitive lipase (HSL) activity when measurements were made on defatted rat heart powders. These lipolytic agents stimulated the activity of this enzyme in a time- and dose-dependent manner. This activation was reversible, because removal of adrenaline from the perfusate was accompanied by the return of type-L HSL activity to control levels. We have reported [Palmer, Caruso & Oscai (1981) Biochem. J. 198, 159-166] that perfusion with low levels of adrenaline, MIX or Bt2 cyclic AMP reduced type-L HSL activity below control levels when measurements were made in aqueous homogenates. However, in the present study, when activities were measured in acetone/diethyl ether heart powders, all concentrations of these agents studied stimulated enzyme activity, and at no concentration was there enzyme inhibition. These data suggest that acetone/diethyl ether treatment may remove a factor that plays a role in type-L HSL regulation. Type-L HSL activity measured in acetone/diethyl ether powders of control and stimulated rat heart exhibited properties that include alkaline pH optimum, serum requirement, activation by heparin and inhibition by high salt and protamine sulphate. These characteristics, in addition to the stability of the enzyme to treatment with organic solvents, fulfil the requirements for the type-L HSL classification.


Salt-Mediated Au-Cu Nanofoam and Au-Cu-Pd Porous Macrobeam Synthesis.
Multi-metallic and alloy nanomaterials enable a broad range of catalytic applications with high surface area and tuning reaction specificity through the variation of metal composition. The ability to synthesize these materials as three-dimensional nanostructures enables control of surface area, pore size and mass transfer properties, electronic conductivity, and ultimately device integration. Au-Cu nanomaterials offer tunable optical and catalytic properties at reduced material cost. The synthesis methods for Au-Cu nanostructures, especially three-dimensional materials, has been limited. Here, we present Au-Cu nanofoams and Au-Cu-Pd macrobeams synthesized from salt precursors. Salt precursors formed from the precipitation of square planar ions resulted in short- and long-range ordered crystals that, when reduced in solution, form nanofoams or macrobeams that can be dried or pressed into freestanding monoliths or films. Metal composition was determined with X-ray diffraction and energy dispersive X-ray spectroscopy. Nitrogen gas adsorption indicated an Au-Cu nanofoam specific surface area of 19.4 m²/g. Specific capacitance determined with electrochemical impedance spectroscopy was 46.0 F/g and 52.5 F/g for Au-Cu nanofoams and Au-Cu-Pd macrobeams, respectively. The use of salt precursors is envisioned as a synthesis route to numerous metal and multi-metallic nanostructures for catalytic, energy storage, and sensing applications.


Signals of recent positive selection in a worldwide sample of human populations.
Genome-wide scans for recent positive selection in humans have yielded insight into the mechanisms underlying the extensive phenotypic diversity in our species, but have focused on a limited number of populations. Here, we present an analysis of recent selection in a global sample of 53 populations, using genotype data from the Human Genome Diversity-CEPH Panel. We refine the geographic distributions of known selective sweeps, and find extensive overlap between these distributions for populations in the same continental region but limited overlap between populations outside these groupings. We present several examples of previously unrecognized candidate targets of selection, including signals at a number of genes in the NRG-ERBB4 developmental pathway in non-African populations. Analysis of recently identified genes involved in complex diseases suggests that there has been selection on loci involved in susceptibility to type II diabetes. Finally, we search for local adaptation between geographically close populations, and highlight several examples.


Sensory noise predicts divisive reshaping of receptive fields.
In order to respond reliably to specific features of their environment, sensory neurons need to integrate multiple incoming noisy signals. Crucially, they also need to compete for the interpretation of those signals with other neurons representing similar features. The form that this competition should take depends critically on the noise corrupting these signals. In this study we show that for the type of noise commonly observed in sensory systems, whose variance scales with the mean signal, sensory neurons should selectively divide their input signals by their predictions, suppressing ambiguous cues while amplifying others. Any change in the stimulus context alters which inputs are suppressed, leading to a deep dynamic reshaping of neural receptive fields going far beyond simple surround suppression. Paradoxically, these highly variable receptive fields go alongside and are in fact required for an invariant representation of external sensory features. In addition to offering a normative account of context-dependent changes in sensory responses, perceptual inference in the presence of signal-dependent noise accounts for ubiquitous features of sensory neurons such as divisive normalization, gain control and contrast dependent temporal dynamics.


Epigenetic analysis of the IFNλ3 gene identifies a novel marker for response to therapy in HCV-infected subjects.
Chronic hepatitis C virus (HCV) infection is characterized by high interindividual variability in response to pegylated interferon and ribavirin. A genetic polymorphism on chromosome 19 (rs12979860) upstream of interferon-λ3 (IFNλ3) is associated with a twofold change in sustained virologic response rate after 48 weeks of treatment with pegylated interferon/ribavirin in HCV genotype 1 (GT1) treatment-naïve patients. We conducted epigenetic analysis on the IFNλ3 promoter to investigate whether DNA methylation is associated with response to HCV therapy. DNA samples from HCV GT1-infected subjects receiving an interferon-free paritaprevir-containing combination regimen (N=540) and from HCV-uninfected, healthy controls (N=124) were analysed for IFNλ3 methylation levels. Methylation was strongly associated with rs12979860 allele status whether adjusting for HCV status (r=65.0%, 95% CI: [60.2%, 69.5%]), or not (r=64.4%), both with P<2.2×10-16 . In HCV GT1-infected subjects, C/C genotypes had significantly lower methylation levels relative to C/T or T/T genotypes (P<1×10-14 ), with each T allele resulting in a nine-unit increase in mean methylation level. Methylation levels did not correlate with response in subjects treated for 12 or 24 weeks. However, non-C/C subjects with higher methylation levels were more likely to relapse when treatment duration was 8 weeks. This analysis suggests that methylation status of the IFNλ3 promoter region may be a useful parameter that identifies patients more likely to relapse following HCV therapy; however, continuing therapy for a sufficient duration can overcome this difference. These findings may provide mechanistic insight into the role of IFNλ3 genetic variants in HCV treatment response.


An iconographic program for computer-controlled whole-cell voltage clamp experiments.
A Macintosh-based system is described for performing instrument control, data acquisition and storage operations in single-electrode whole-cell voltage clamp experiments. The system consists of a commercially available voltage clamp amplifier, multifunction input/output (I/O) board, graphical programming language (LabVIEW 2) and custom built 'virtual instrument' (VI). The I/O board is capable of fast (up to 110 ksample/s) multichannel analog-to-digital (A/D) conversion with 12-bit resolution. It can control the gain settings of the clamp unit through digital I/O lines and generate the P/N leak subtraction protocol to eliminate the linear portion of capacitive currents using analog output voltages and gating pulses. Complete voltage clamp protocols can be implemented using the on-screen front panel controls of the VI. It enables the user to visualize the acquired data, to graph sets of current-voltage (I-V) relations or to fit single-exponential functions to one current trace. To evaluate the adequacy of whole-cell recording, the total membrane capacitance (Cm), the series resistance (Rs) and the time constant (tau c) of the decay of the capacitive current are calculated using the single-exponential function fit to the data. The system is particularly well suited to the study of large quantities of transmembrane I-V relationships. Source code for the crucial elements of the VI as well as sample recordings from a cultured spinal cord neuron, illustrating system operation, are presented.


Drainage Tube Placement for the Management of Rebleeding After Vascular Embolization in a Patient With Neurofibromatosis Type 1.
Neurofibromatosis type 1 (NF-1), a hereditary disease caused by a mutation of a gene on chromosome 17q11.2, is associated with manifestations in several organs. Although infrequent, vascular abnormalities are a complication of NF-1, and they are the second most common cause of death in patients with NF-1. Repairing the nutrient artery and achieving hemostasis are difficult once the artery has failed, thereby resulting in poor treatment outcomes. Herein, we report a case of a patient with NF-1 who presented with an enormous cervical hematoma caused by bleeding from a branch of the external carotid artery. Vascular embolization was performed initially; however, rebleeding from the embolized site occurred. Following the removal of the hematoma, drainage tube placement was effective in preventing micro-bleeding. Thus, drainage tube placement may be an effective treatment option in patients with rebleeding.


Selective hypoaldosteronism: a study of steroid biosynthetic pathways under adrenocorticotrophin and angiotensin II infusion.
1. The functional integrity of the adrenal cortex has been tested in a case of selective hypoaldosteronism by adrenocorticotrophin (ACTH) and angiotensin II (AII) infusion. 2. During ACTH infusion a normal functioning zona fasciculata was indicated by the impressive increase of the ACTH-dependent plasma steroids; the aldosterone response was moderate. 3. During AII infusion the plasma aldosterone response was blunted with an unexpected dose-dependent increase in pregnenolone, resulting in abnormal decreasing progesterone/pregnenolone ratios during the infusion, suggesting a slow-down in the conversion of pregnenolone into progesterone. 4. This defect, a probable consequence of chronic renin deficiency on the zona glomerulosa, could be a contributing factor to the hypoaldosteronism.


A deep matrix completion method for imputing missing histological data in breast cancer by integrating DCE-MRI radiomics.
Clinical indicators of histological information are important for breast cancer treatment and operational decision making, but these histological data suffer from frequent missing values due to various experimental/clinical reasons. The limited amount of histological information from breast cancer samples impedes the accuracy of data imputation. The purpose of this study was to impute missing histological data, including Ki-67 expression level, luminal A subtype, and histological grade, by integrating tumor radiomics. To this end, a deep matrix completion (DMC) method was proposed for imputing missing histological data using nonmissing features composed of histological and tumor radiomics (termed radiohistological features). DMC finds a latent nonlinear association between radiohistological features across all samples and samples for all the features. Radiomic features of morphologic, statistical, and texture were extracted from dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) inside the tumor. Experiments on missing histological data imputation were performed with a variable number of features and missing data rates. The performance of the DMC method was compared with those of the nonnegative matrix factorization (NMF) and collaborative filtering (MCF)-based data imputation methods. The area under the curve (AUC) was used to assess the performance of missing histological data imputation. By integrating radiomics from DCE-MRI, the DMC method showed significantly better performance in terms of AUC than that using only histological data. Additionally, DMC using 120 radiomic features showed an optimal prediction performance (AUC = 0.793), which was better than the NMF (AUC = 0.756) and MCF methods (AUC = 0.706; corrected p = 0.001). The DMC method consistently performed better than the NMF and MCF methods with a variable number of radiomic features and missing data rates. DMC improves imputation performance by integrating tumor histological and radiomics data. This study transforms latent imaging-scale patterns for interactions with molecular-scale histological information and is promising in the tumor characterization and management of patients.


Urinary Amino-Polycyclic Aromatic Hydrocarbons in Urban Residents: Finding a Biomarker for Residential Exposure to Diesel Traffic.
Despite substantial evidence of marked exposure to and ill-health effects from diesel exhaust (DE) emissions among occupational population (e.g., miners, truck drivers, and taxi drivers), it is less understood to what extent non-occupational population was exposed to DE among various combustion sources, largely due to the lack of biomarkers that would indicate specific exposure to DE. We evaluated whether urinary amino-polycyclic aromatic hydrocarbons (APAHs), such as major metabolites of DE-specific nitrated PAHs, can be used as DE exposure biomarkers in residential settings. We measured five urinary APAHs in 177 urine samples from 98 UK residents, 89 (91%) of them were London residents, and estimated their residential proximity to various traffic indicators (e.g., the road type, road length, traffic flow, and traffic volume). Participants living within 100 m of major roads exhibited increased levels of all five APAHs, among which 2-amino-fluorene (2-AFLU) reached statistical significance (p < 0.05). We estimated that a 10 m increase in the length of nearby major roads (<100 m) was associated with a 4.4% (95% CI of 1.1 to 7.6%) increase in 2-AFLU levels. Levels of 2-AFLU were significantly associated with the traffic flow of nearby buses and heavy-duty vehicles but not motorbikes, taxis, or coaches. We did not observe a significant association between distance to major roads or the sum of the major road length within 100 m with the other four biomarker concentrations. These results suggest the use of urinary 2-AFLU as a biomarker of DE exposure in urban residents.