ZnO, AgCl and AgCl/ZnO nanocomposites incorporated chitosan in the form of hydrogel beads for photocatalytic degradation of MB, E. coli and S. aureus.
Significant improvement of effective and low-cost decolorization and disinfecting technologies is required to address the problems created by dyes and dangerous microorganisms from water and wastewaters. This article expresses the degradation of methylene blue (MB), Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) as gram negative and positive bacteria via a chitosan/AgCl/ZnO (CS/AgCl/ZnO) nanocomposite hydrogel beads system as a photocatalyst under visible light irradiation. The techniques such as FT-IR, SEM, EDAX, TGA, and XRD were applied to recognize the synthesized beads. Decolorization and disinfection experimental results revealed that the hydrogel beads system effectively degrade MB and bacteria. Also, the effects of the initial amount of catalysts, pH, coions and initial concentration of dye on the photocatalytic decolorization were investigated. Moreover, kinetics analysis indicates that the photocatalytic degradation rate of MB and bacteria can be described by Langmuir-Hinshelwood (L-H) and Weibull inactivation models, respectively. We provide a reusable and recoverable effective organic/inorganic photocatalyst in the form of beads that could solve the disadvantages of powder photocatalytic, without reducing the efficiency.


Encounter with unexpected collagenase-1 selective inhibitor: switchover of inhibitor binding pocket induced by fluorine atom.
Phosphonamide-based inhibitors having trifluoromethyl moiety showed highly selective inhibition against MMP-1. A possible mechanism of the selectivity of MMP-1 inhibitors through the switchover of the binding pocket was speculated by computational calculations. As a consequence of the unexpected selectivity, the specific interaction of CF3 group of the inhibitor and Arg214 in the S1' pocket of MMP-1 conducted a low binding energy.


Three-site sandwich radioimmunoassay with monoclonal antibodies for a sensitive determination of human alpha-fetoprotein.
Utilizing monoclonal antibodies against human alpha-fetoprotein, 3 distinct antigenic determinants were identified. These antigenic determinants, provisionally designated a, b and c, were arranged in such a manner that the binding of one determinant with the corresponding antibody did not inhibit, or only barely inhibited the binding of antibodies directed to the other 2 determinants. Monoclonal antibodies with 3 different specificities were, therefore, applied to develop a sandwich-type solid-phase radioimmunoassay of the antigen in which wells were coated with anti-a, and radiolabeled anti-b together with radiolabeled anti-c was employed to detect the bound antigen. The 3-site sandwich radioimmunoassay involving 3 different determinants gave a higher sensitivity than 2-site assays in which only anti-b or anti-c was employed as a radiolabeled reagent, because the radioactivity of the 2 labeled antibodies was added on the antigen bound to immobilized anti-a.


Facilitated transport of oxygen through hemoglobin solutions.
A method for quantitative analysis of hemoglobin color to estimate the oxygen saturation was developed. The method uses an image-input and -processing system composed of a 3-tube video camera and a digital image analyzer. When the system is connected to a microscope, the facilitated diffusion of oxygen in hemoglobin solutions can be observed and analyzed in a position-sensitive manner. The results led to a new transport model expressed as: J = k'CY/d, where J is the oxygen flux, C is the hemoglobin concentration, Y is the oxygen saturation of hemoglobin, d is the distance between the entrance of oxygen and the diffusion front, and k' is a constant.


Compartmentalization of calmodulin and tubulin in the male C57BL/6J mouse brain: heterogeneity of age changes in calmodulin compartments.
Calmodulin (CaM) and tubulin were analyzed by radioimmunoassay in subcellular fractions prepared from cerebral cortex and striatum of aging male C57BL/6J mice. Three fractions were prepared by a new procedure: cytosol (soluble); EGTA-releasable, membrane-bound; and detergent-extractable (Triton X-100), membrane-bound fractions. CaM concentration in all three fractions prepared from striatum showed small (10-15%, p less than 0.05) decreases with age (3-31 months). Cortical CaM concentrations were less affected by age, and only the EGTA-releasable fraction decreased. To compare functional activity and immunoreactivity of CaM, soluble CaM was also assayed by the activation of cyclic nucleotide phosphodiesterase (PDE). The radioimmunoassay and PDE activation assays gave equivalent results, suggesting that no alteration occurred with age in biological activity of CaM, via post-translational modification or other mechanisms. Soluble and particulate tubulin concentration did not change significantly with age in either brain region. The changes observed in striatal CaM, particularly in membrane-bound compartments, could contribute to the age-related decline in mammalian basal ganglial function.


Effects of zero-filling and apodization on spectral integrals in discrete Fourier-transform spectroscopy of noisy data.
The influence of noise on the standard deviation of spectral integrals is examined. Calculations assuming discrete Fourier-transform data are compared with Monte-Carlo simulations. The effects of zero-filling and apodization are examined for free-induction-decay (FID) signals and for symmetric spin-echo signals in one and two dimensions, with particular attention to features not previously presented in the literature. Findings suggest that for mild apodization, the known sensitivity enhancement due to zero-filling in either the real or the imaginary part signal [E. Bartholdi, R.R. Ernst, Fourier spectroscopy and the causality principle, J. Magn. Reson., 11 (1973) 9-19] is maintained; however, for stronger apodization filters, this enhancement can be obliterated completely. It is shown that results obtained by analysis of one-dimensional signals can be readily applied to multi-dimensional data. Furthermore, zero-filling has a negligible effect for symmetric spin-echo signals with implications for signal averaging in magnetic resonance imaging and spectroscopic imaging.


Effects of end-effector robot-assisted gait training on gait ability, muscle strength, and balance in patients with spinal cord injury.
There is no randomized controlled study about the effects of end-effector robot-assisted gait training (RAGT) in patients with spinal cord injury (SCI). To examine the effects of end-effector RAGT on gait and balance abilities in SCI. Thirty-one patients were randomly assigned to the RAGT (Morning Walk®, Curexo, Seoul, South Korea) or conventional therapy (CT) group. Patients were assessed using the 10-meter walk test (10MWT), 6-minute walk test (6mWT), lower extremity motor score (LEMS) and proprioception, Berg Balance Scale (BBS), Walking Index for Spinal Cord Injury-II (WISCI-II), and mobility category of Spinal Cord Independence Measure-III. All clinical outcome measures significantly improved in both groups. The BBS and WISCI-II were significantly improved in the RAGT group compared to the CT group. In the RAGT group, pre-LEMS and pre-WISCI-II of the 10MWT improved group and pre-BBS of the 6mWT improved group were higher than those of the 10MWT non-improved and 6mWT non-improved group, respectively. End-effector RAGT and CT in patients with incomplete SCI could lead to improvements in gait ability, lower extremity muscle strength, balance, proprioception, and mobility. Additionally, end-effector RAGT could improve balance and gait abilities substantially better than CT.


Has the raphe dorsalis nucleus an asymmetric function?
Differential pulse voltammetry (DPV) was used to measure 5-hydroxyindoles in the striata of anaesthetized rats. The amplitude of the voltammetry peaks in both striata decreased when 5-hydroxytryptophan (5-HTP) or 5-hydroxytryptamine (5-HT) was injected into the left side of the raphe dorsalis nucleus (RDN), but increased when either substance was injected into the right side. These findings indicate that the 5-hydroxyindole based communication between the striata and the RDN has a lateral asymmetry. The data are discussed with reference to histological observations suggesting a higher density of 5-hydroxyindole containing cell bodies in the lateral region than in the midline of this "unpaired" nucleus.


Novel approach to target cancer stem cells for therapy.
Even though lots of efforts have been made to find different strategies for cancer treatments, currently available therapeutic approaches are chemotherapy, radiation and surgery or combination of these. These treatments prolonged the survival of patients but did not assure complete cure of the disease. Recent scientific evidences suggest that cancer stem cells (CSC) are responsible for recurrence, resistance and existence of this disease even after various therapeutic treatments. Therefore, we hypothesize that the best approach is to target CSCs along with cancer cells for complete remission of the disease. Before targeting these cells, studying their morphological, proliferation, behavioral aspects, physico-chemical interaction and characterizations are very important. For therapeutic approach the differentiation capacity of these cells to cancer cells with or without drugs is critical. To study basic parameters; the best approach would be aseptic sorting of CSCs from cancer cells based on specific cell surface markers by flowcytometer or magnetic cell sorter. The sorted cells have to be grown in culture conditions and treat with optimum concentrations of drugs to target CSC and cancer cell to find appropriate potential combination.


Soluble AXL is a novel blood marker for early detection of pancreatic ductal adenocarcinoma and differential diagnosis from chronic pancreatitis.
Early diagnosis is crucial for patients with pancreatic ductal adenocarcinoma (PDAC). The AXL receptor tyrosine kinase is proteolytically processed releasing a soluble form (sAXL) into the blood stream. Here we explore the use of sAXL as a biomarker for PDAC. AXL was analysed by immunohistochemistry in human pancreatic tissue samples. RNA expression analysis was performed using TCGA/GTEx databases. The plasma concentrations of sAXL, its ligand GAS6, and CA19-9 were studied in two independent cohorts, the HMar cohort (n = 59) and the HClinic cohort (n = 142), including healthy controls, chronic pancreatitis (CP) or PDAC patients, and in a familial PDAC cohort (n = 68). AXL expression and sAXL release were studied in PDAC cell lines and murine models. AXL is increased in PDAC and precursor lesions as compared to CP or controls. sAXL determined in plasma from two independent cohorts was significantly increased in the PDAC group as compared to healthy controls or CP patients. Patients with high levels of AXL have a lower overall survival. ROC analysis of the plasma levels of sAXL, GAS6, or CA19-9 in our cohorts revealed that sAXL outperformed CA19-9 for discriminating between CP and PDAC. Using both sAXL and CA19-9 increased the diagnostic value. These results were validated in murine models, showing increased sAXL specifically in animals developing PDAC but not those with precursor lesions or acinar tumours. sAXL appears as a biomarker for early detection of PDAC and PDAC-CP discrimination that could accelerate treatment and improve its dismal prognosis. This work was supported by grants PI20/00625 (PN), RTI2018-095672-B-I00 (AM and PGF), PI20/01696 (MG) and PI18/01034 (AC) from MICINN-FEDER and grant 2017/SGR/225 (PN) from Generalitat de Catalunya.


Fundamental role of Lactobacillus plantarum and inulin in improving safety and quality of Karish cheese.
Karish cheese manufactured traditionally from raw milk may harbor many biological health hazards. Production of safe pasteurized Karish cheese with improved sensory characteristics using probiotics and prebiotics (synbiotic Karish cheese). Laboratory Karish cheese was made to study the effect of Lactobacillus plantarum with and without inulin on cheese quality. Treatments were examined for sensory, chemical, and microbial quality, shelf life, and survival of L. plantarum were also monitored. The antimicrobial effect of L. plantarum and inulin against Enterobacter aerogenes in cheese was evaluated. Sensory, chemical, and microbial quality of Karish cheese supplemented with L. plantarum and inulin were positively affected; moreover, the shelf life was extended up to 28 days. Karish cheese contained L. plantarum showed the highest flavor score, while treatment contained both L. plantarum and inulin attained the best body and texture score. Moreover, L. plantarum and inulin significantly reduced E. aerogenes count during Karish cheese chilled storage; the reduction log reached 3.76 log10cfu/g on the seventh day of storage compared to control. Additionally, Inulin significantly increased the survival of L. plantarum throughout the storage period. This study concluded that using probiotics and prebiotics in Karish cheese synergistically improved its sensory properties, safety, and hygienic quality.


A comparison of alcohol and drug use patterns of homosexual and heterosexual men: the San Francisco Men's Health Study.
Very high prevalence rates for problem drinking and/or alcoholism have been estimated for homosexual male populations. Populations characterized by high prevalence rates for problematic drug or alcohol use are of special interest since study of such groups may help to provide insights regarding the processes associated with problematic substance use. However, prevalence estimates for problematic substance use within gay male populations have most typically relied on convenience samples and generally contain an over-representation of bar patrons. This study reports data from a large scale random household sample of homosexual and heterosexual men who live in an urban district of San Francisco, California. Few differences were noted in the drinking patterns of these men, although important differences were found in the prevalence of drug use over a 6-month period. Differences in the prevalence of at least weekly drug use were comparatively minor, however. Health policy implications of these findings are discussed.


Inhibitory potential of naphthoquinones leached from leaves and exuded from roots of the invasive plant Impatiens glandulifera.
Exploring the effects of allelopathic plant chemicals on the growth of native vegetation is essential to understand their ecological roles and importance in exotic plant invasion. Naphthoquinones have been identified as potential growth inhibitors produced by Impatiens glandulifera, an exotic annual plant that recently invaded temperate forests in Europe. However, naphthoquinone release and inhibitory potential have not been examined. We quantified the naphthoquinone content in cotyledons, leaves, stems, and roots from plants of different ages of both the invasive I. glandulifera and native Impatiens noli-tangere as well as in soil extracts and rainwater rinsed from leaves of either plant species by using ultra-high pressure liquid chromatography-mass spectrometry (UHPLC-MS). We identified the compound 2-methoxy-1,4-naphthoquinone (2-MNQ) exclusively in plant organs of I. glandulifera, in resin bags buried into the soil of patches invaded by I. glandulifera, and in rainwater rinsed from its leaves. This indicates that 2-MNQ is released from the roots of I. glandulifera and leached from its leaves by rain. Specific bioassays using aqueous shoot and root extracts revealed a strong inhibitory effect on the germination of two native forest herbs and on the mycelium growth of three ectomycorrhiza fungi. These findings suggest that the release of 2-MNQ may contribute to the invasion success of I. glandulifera and support the novel weapons hypothesis.


Transuranic nuclides Pu, Am and Cm isotopes, and 90Sr in seafloor sediments off the Fukushima Daiichi Nuclear Power Plant during the period from 2012 to 2019.
The 238Pu, 239+240Pu, 241Am, 242Cm, 243+244Cm and 90Sr concentrations in seafloor surface sediments collected at three sampling stations off the Fukushima Daiichi Nuclear Power Plant (FDNPP) site during the period from 2012 to 2019 were determined to elucidate the impacts of the FDNPP accident onto their concentrations in coastal sediments and to discuss the sources of the measured radionuclides. The 239+240Pu, 241Pu and 241Am concentrations and 240Pu/239Pu atom ratios in a sediment core were also determined to allow comparison of their inventories between this study and previously reported values and to identify the Pu sources. The 238Pu, 239+240Pu, 241Am and 90Sr concentrations showed no remarkable temporal variations; no significant increases in concentrations after the FDNPP accident were observed; these concentrations were comfortably within the previously reported concentration range; and no detectable 242Cm and 243+244Cm amounts were observed in surface sediments. The observed 238Pu/239+240Pu activity ratios were approximately two orders of magnitudes lower than those for the damaged FDNPP reactor core inventories and the observed values in terrestrial samples after the accident. The 239+240Pu, 241Pu and 241Am inventories in the sediment core were 389 ± 5, 503 ± 33 and 214 ± 3 Bq m-2, respectively. The 239+240Pu inventory was about an order of magnitude greater than the expected cumulative deposition density of global fallout from atmospheric nuclear weapons testing due to an enhanced scavenging effect. The 240Pu/239Pu atom ratios in the sediment core ranged from 0.239 to 0.246 with a mean value of 0.242 ± 0.002; these ratios were clearly greater than the mean global fallout ratio of 0.18. The results for 238Pu/239+240Pu activity ratios and 240Pu/239Pu atom ratios reflected a mixture of global fallout and Pacific Proving Grounds (PPG) close-in fallout Pu rather than Fukushima accident-derived Pu. The sediment column inventory for 239+240Pu originating from the PPG close-in fallout was calculated as 166 Bq m-2, which corresponded to 43% of the total inventory. A significant amount of the PPG-derived Pu has been transported by ocean currents and then preferentially scavenged in the coastal waters of Japan.


Multiple fibroepithelial hyperplasias of the oral mucosa.
Reported is a rare case of multiple fibroepithelial hyperplasias of the oral cavity, and the suggestion is made that several previous authors may have confused this lesion with focal epithelial hyperplasia (Heck's disease). The differential diagnosis of multiple fibroepithelial hyperplasias is discussed in order to elucidate some of the confusions in the diagnosis of cases of multiple papules and nodules of the oral mucosa.


Insulin resistance and cardiovascular disease risk factors in children of parents with the insulin resistance (metabolic) syndrome.
To evaluate whether children of parents with the insulin resistance syndrome (IRS) themselves have greater insulin resistance and unfavorable patterns of cardiovascular disease (CVD) risk factors. This cross-sectional study included 220 white and 36 black children aged 11-15 years identified through a school-based blood pressure screening program, along with 378 of their parents. Measures of insulin resistance (glucose disposal per minute per kilogram of lean body mass in a euglycemic-hyperinsulinemic clamp [Mlbm] and fasting insulin), adiposity, and other CVD risk factors were compared in children with and without a parental history of IRS, defined according to the National Cholesterol Education Program Adult Treatment Panel III consensus definition. Compared with children in whom neither parent had IRS, children who had at least one parent with the syndrome had statistically significantly lower mean Mlbm (12.1 vs. 13.6 mg.kg(-1).min(-1); P=0.04) and higher fasting insulin (geometric means 99 vs. 76 pmol/l; P=0.01) after adjustment for sex, race, age, and Tanner stage. Mean BMI, waist circumference, waist-to-hip ratio, triceps and subscapular skinfolds, and percentage of body fat were also significantly higher in children of an affected parent, but there were no significant differences in lipid or blood pressure levels between the two groups. Insulin resistance and obesity may be the earliest manifestations of IRS in children with a parental history of the syndrome.


Primary structure of T4 DNA polymerase. Evolutionary relatedness to eucaryotic and other procaryotic DNA polymerases.
Bacteriophage T4 gene 43 codes for the viral DNA polymerase. We report here the sequence of gene 43 and about 70 nucleotides of 5'- and 3'-flanking sequences, determined by both DNA and RNA sequencing. We have also purified T4 DNA polymerase from T4 infected Escherichia coli and from E. coli containing a gene 43 overexpression vector. A major portion of the deduced amino acid sequence has been verified by peptide mapping and sequencing of the purified DNA polymerase. All these results are consistent with T4 DNA polymerase having 898 amino acids with a calculated Mr = 103,572. Comparison of the primary structure of T4 DNA polymerase with the sequence of other procaryotic and eucaryotic DNA polymerases indicates that T4 DNA polymerase has regions of striking similarity with animal virus DNA polymerases and human DNA polymerase alpha. Surprisingly, T4 DNA polymerase shares only limited similarity with E. coli polymerase I and no detectable similarity with T7 DNA polymerase. Based on the location of specific mutations in T4 DNA polymerase and the conservation of particular sequences in T4 and eucaryotic DNA polymerases, we propose that the NH2-terminal half of T4 DNA polymerase forms a domain that carries out the 3'----5' exonuclease activity whereas the COOH-terminal half of the polypeptide contains the dNTP-binding site and is necessary for DNA synthesis.


Protection of hematopoietic cells against combined O6-benzylguanine and chloroethylnitrosourea treatment by mutant forms of O6-methylguanine DNA methyltransferase.
This report demonstrates that expression of the P140A O6-methylguanine DNA methyl transferase (MGMT) mutant via retrovirus-mediated gene transfer leads to significant, but modest, resistance of cells to both 6-benzylguanine (6-BG) depletion and treatment with 1,3-bis (2-chloroethyl)-1-nitrosourea (BCNU). Expression of the P140A/G156A double mutant appeared to be associated with reduced or unstable protein in hematopoietic cells.


Chlorpyrifos induces placental oxidative stress and barrier dysfunction by inducing mitochondrial apoptosis through the ERK/MAPK signaling pathway: In vitro and in vivo studies.
Chlorpyrifos (CPF) is an organophosphorus pesticide that is widely used in agricultural production and residential environments worldwide. In this study, we determined the harmful effects and toxicological mechanism of CPF in porcine trophectoderm (pTr) cells and the placenta of female mice during pregnancy. The findings revealed that CPF significantly decreased cell viability and increased intracellular lactate dehydrogenase (LDH) release in pTr cells. Similarly, CPF induced reproductive toxicity in pregnant maternal mice, including decreased maternal, fetal, and placental weights. Moreover, following CPF treatment, pTr cells and the placenta of female mice showed significant apoptosis. JC-1 staining and flow cytometry analysis also revealed that the mitochondrial membrane potential (MMP) of pTr cells treated with CPF was significantly depolarized. Additionally, CPF can induce an increase in reactive oxygen species (ROS) and barrier dysfunction in pTr cells and the placenta of female mice. We further verified that CPF-induced mitochondrial apoptosis is mediated by the MAPK signaling pathway, as shown by using of small molecular inhibitors of related proteins. Also, CPF-induced oxidative stress, barrier dysfunction, and mitochondrial apoptosis in pTr cells were alleviated by U0126, an inhibitor of the ERK/MAPK signaling pathway. These findings suggested that exposure to CPF in early pregnancy might be a potential risk fator affecting placental formation and function in humans and animals.


Expression and distribution of cell adhesion-related proteins in bovine parthenogenetic embryos: The effects of oocyte vitrification.
The objective was to investigate expression of cell adhesion-related proteins (E-cadherin, β-catenin, and the cytoskeletal protein F-actin) in bovine parthenogenetic embryos derived from vitrified-warmed oocytes. Bovine oocytes at metaphase II were randomly allocated into three groups: (1) untreated (control); (2) exposed to vitrification solution without freezing (toxicity); and (3) vitrified and warmed by the open-pulled straw method (vitrification). After parthenogenetic activation, in the vitrification group compared with the control, the timing of compaction was delayed in (108-120 vs. 96-108 hours, respectively), and the percentage of blastocysts that developed from eight-cell embryos was lower (32.08% vs. 61.03%; P < 0.05). To investigate whether vitrification delayed embryo compaction by affecting adhesion junction formation and function, immunostaining and quantitative reverse transcription polymerase chain reaction were done to characterize distribution patterns (E-cadherin, β-catenin, and the cytoskeletal protein F-actin) and expression levels of cell adhesion-related proteins (β-catenin). Distribution of β-catenin in eight-cell embryos from the vitrification group changed dramatically compared with the control and toxicity groups. Relative expression of β-catenin at the mRNA and protein levels was lower (P < 0.05) than that of the fresh and toxicity groups. However, expression and distribution of E-cadherin were similar among groups. In conclusion, abnormal distribution and decreased expression of β-catenin in bovine parthenogenetic eight-cell embryos derived from vitrified-warmed oocytes were associated with embryo compaction and reduced competence for subsequent embryo development.


The impact of COVID-19 on the intention of third-child in China: an empirical analysis based on survey data.
Against the grim background of declining intention to have children, the ravages of COVID-19 have pushed China and the world into a more complex social environment. To adapt to the new situation, the Chinese government implemented the three-child policy in 2021. COVID-19 pandemic indirectly affects the country's internal economic development, employment, fertility plans or intention, and other major issues related to the people's livelihood, while undermining the stable operation of society. This paper explores the question that will COVID-19 pandemic affect Chinese people's intention to have a third child. And What are the relevant factors inside? The data in this paper are from the Survey released by the Population Policy and Development Research Center of Chongqing Technology and Business University (PDPR-CTBU), including 10,323 samples from mainland China. This paper uses the logit regression model and KHB mediated effect model (a binary response model given by Karlson, Holm, and Breen) to investigate the impact of the COVID-19 pandemic and other factors on Chinese residents' intention to have a third child. The results suggest that the COVID-19 pandemic has a negative effect on Chinese residents' intention to have a third child. In-depth research on the mediating effect of KHB shows that COVID-19 pandemic will further inhibit residents' intention to have a third child by affecting their childcare arrangements, increasing their childcare costs, and increasing their exposure to occupational hazards. This paper is more pioneering in focusing on the impact of the COVID-19 epidemic on the intention to have three children in China. The study provides empirical evidence for understanding the impact of COVID-19 epidemic on fertility intentions, albeit in the context of policy support.


Prevention of Retrograde Ascending Aortic Dissection by Cardiac Pacing During Hybrid Surgery for Zone 0 Aortic Arch Repair.
Retrograde type A dissection (RTAD) after zone 0 hybrid aortic arch repair is highly lethal and not infrequent complication. The aim of this study was to assess the safety and effectiveness of rapid cardiac pacing as an adjunctive tool to prevent RTAD during or after hybrid procedures for zone 0 disease. We performed a retrospective review of 42 consecutive patients with zone 0 hybrid aortic arch repair between November 2004 and January 2018. Right ventricular pacing was carried out through unipolar electrodes attached to the epicardium of the right ventricle through the sternotomy (the indifferent electrode was in the subcutaneous tissue). Pacing was utilised during the clamping of the ascending aorta, release of the aortic clamp, and stent-graft deployment. Operative indications were aortic arch aneurysm 45% (n = 19), aortic arch dissection 45% (n = 19), traumatic rupture of isthmus 7% (n = 3), and type IA endoleak 2% (n = 1). Urgent procedures 48% (n = 20). The mean proximal aortic diameter was 34.14 ± 2.9 mm. Mean stent-graft oversizing was 12.97 ± 3.4%. The 30-day mortality rate was 14% (n = 6). RTAD was observed in 7% (n = 3). The actuarial survival rate was 74% over a mean follow-up of 50 ± 30.2 months. Since January 2013, rapid right ventricular pacing (overdrive pacing at a rate of 200 beats/min) was systematically used (n = 24). No RTAD was observed in this group of patients. Rapid right ventricular pacing reduced significatively the risk of RTAD (P = 0.038). Rapid right ventricular pacing is an effective method of inducing hypotension and appears to decrease the risk of retrograde type A dissection after zone 0 hybrid aortic arch repair.


A performance study of a wearable balance assistance device consisting of scissored-pair control moment gyroscopes and a two-axis inclination sensor.
Excessive postural sway while standing can lead to falls and injuries. A designed wearable balance assistance device which consists of scissored-pair control moment gyroscopes and a two-axis inclination sensor is introduced to reduce fall risk from excessive sway among the elderly. The prototype has dimensions of H50cm × W44cm × D30cm and weighs 15.03 kg. This study aims to investigate the effects of generated torque of the prototype on human subjects and aims to determine if the two-axis inclination sensor can detect sway amplitude and sway direction during an occurrence of excessive sway. Two healthy male subjects participated in the study. According to the results, the detected body incline angle related to the acquired sway amplitude of COP trajectories with correlation factors of 0.92 and 0.88 for the two subjects. The detected sway angle related to the acquired sway direction of COP trajectories with the correlation factors of 0.99 and 0.98 for the two subjects. The maximum-allowable generated torque of the prototype with an assigned actuating angle varying within ±15.6° from the acquired sway direction of COP trajectories was able to drive the COP of 60-kilogram-weighted healthy subject maintaining balance at posterolateral limits of stability with an average body incline angle of 5.74° to pass his standing secure zone. The results indicate that the prototype has the potential of being a wearable balance assistance device which can reduce fall risk from excessive sway among the elderly; however, some improvements are still required in regards to shape, size, mass, generated torque, and strength.


Cardiovascular risk reduction: the future of cholesterol lowering drugs.
Atherosclerotic cardiovascular disease is the leading cause of death in developed and developing countries. LDL lowering therapies have a major role in reduction of cardiovascular events. Statins have been the mainstay of LDL lowering therapies with 20-60% reductions in LDL cholesterol. Monoclonal antibodies to proprotein convertase subtilisin kexin type 9 (PCSK9) represent a new therapeutic option, reducing LDL cholesterol by an additional 40-70% on top of other lipid lowering therapies. This is likely to produce significant cardiovascular risk reduction, although clinical cardiovascular outcomes trials are still in progress. HDL cholesterol raising and triglyceride lowering therapies have not yet shown unequivocal benefits for cardiovascular risk reduction. New therapies in these areas are in development, and their future promise remains to be demonstrated.


Impact of structured report on the quality of preoperative CT staging of pancreatic ductal adenocarcinoma: assessment of intra- and inter-reader variability.
To evaluate whether a structured radiology report improves the completeness of preoperative CT staging of pancreatic ductal adenocarcinoma (PDA) compared to conventional free-text reports. We retrospectively included 27 patients (mean age, 64 ± 11.1 years) referred for pancreatic preoperative CT scan for staging of PDA between 2015 and 2018 and in whom a diagnosis of pancreatic adenocarcinoma was ultimately confirmed. Four readers independently reported CT scans with both conventional free-text and structured reports. Differences in reported morphologic and vascular features with the two reports were assessed through McNemar Test. Intra-reader and inter-reader were calculated. A total of 216 reports were completed by four different readers including 108 free-text and 108 structured reports. Overall, 139 of 540 morphologic characteristics of PDA and 869 of 1188 vascular key features were only described in structured reports. Encasement of left gastric artery, gastroduodenal artery and splenic artery was described in up to 14.8% using free-text reports and in up to 29.6% using structured report, resulting in low-intra-reader agreement (k = 0.033-0.216). Inter-reader agreement improved with structured report compared to free-text one for left gastric artery (ICC = 0.844 vs. ICC = 0.493, respectively), gastroduodenal artery (ICC = 0.730 vs. ICC = 0.449, respectively), portal vein (ICC = 0.847 vs. ICC = 0.638, respectively), portal confluence (ICC = 0.848 vs. ICC = 0.422, respectively) superior mesenteric vein (ICC = 0.765 vs. ICC = 0.695, respectively), and splenic vein (ICC = 0.921 vs. ICC = 0.841, respectively). Structured reports for PDA staging significantly reduces the number of missing morphological and vascular features of PDA and improves the inter-reader agreement compared to free-text reports.


Microstructural Evolution in 2101 Lean Duplex Stainless Steel During Low- and Intermediate-Temperature Aging.
The microstructural evolution of a 2101 lean duplex stainless steel (DSS) during isothermal aging from room temperature to 470 °C was investigated using thermoelectric power (TEP) measurements to follow the kinetics, atom probe tomography, and transmission electron microscopy. Despite the low Ni, Cr, and Mo contents, the lean DSS was sensitive to α-α' phase separation and Ni-Mn-Si-Al-Cu clustering at intermediate temperatures. The time-temperature pairs characteristic of the early stages of ferrite decomposition were determined from the TEP kinetics. Considering their composition and locations, the clusters are most likely G phase precursors.


Influence of dextran polymer particles (Sephadex G-200) on the chemiluminescence response of murine peritoneal leukocytes.
Murine peritoneal leukocytes exhibited enhanced levels of chemiluminescence (Cl) when derived from mice inoculated intraperitoneally with Sephadex G-200 gel. This could be observed in the background, 'spontaneous' Cl reactivity, and in Cl elicited by in vitro addition of zymosan or mitogens. Leukocytes found in the peritoneum after Sephadex G-200 inoculation are predominantly neutrophils. It is suggested that the enhanced Cl reactivity is due to activation and proliferation of neutrophils.


Chlorantholides A-F, eudesmane-type sesquiterpene lactones from Chloranthus elatior.
Six eudesmane-type sesquiterpene lactones, named chlorantholides A-F, were isolated from the ethanol extract of Chloranthus elatior (Chloranthaceae) together with 12 known compounds. Their structures were elucidated on the basis of extensive spectroscopic analysis, and their absolute configurations were studied by the CD exciton chirality method. The structure of a recently reported eudesmanolide from Chloranthus anhuiensis: 8β-hydroxy-1-oxoeudesma-3,7(11)-dien-12,8-olide, was also revised as 8β-hydroxy-2-oxoeudesma-3,7(11)-dien-12,8-olide (chlorantholide D).


Sick day management in people with chronic kidney disease: a scoping review.
Use of certain medications during an acute illness may put patients at an increased risk of acute kidney injury (AKI). Patients with chronic kidney disease (CKD) are at higher risk of developing superimposed AKI. The aim of this scoping review is to collate and characterise existing evidence on sick day management considerations and practices during acute illness in people with CKD. We searched Embase, CINAHL, MEDLINE, International Pharmaceutical Abstract, Scopus, Google Scholar and grey literature sources. We followed the methodological framework for scoping reviews, while information was extracted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for scoping reviews. Findings are presented thematically. Ten studies and seven guidelines met the inclusion criteria. Studies were targeted at patients, general practitioners, pharmacists, and nurses. The major themes identified included development and feasibility testing of a sick day management protocol, current practice of temporary medication discontinuation, and outcomes. Most guidelines provided recommendations for sick day management largely based on expert consensus. A digital intervention was deemed highly acceptable and easy to use, whereas patient handouts were more effective when provided along with dialogue with a health professional. While there is little evidence on the impact of sick day protocols on outcomes, a single randomised trial reported no significant association between sick day protocols and change in kidney function, AKI incidents or risk of hospitalisation. The nascent literature on sick day management in patients with CKD revealed the limited available evidence to provide guidance on implementation and on outcomes. Future research needs to clarify sick day recommendations and assess their impact on clinical outcomes including prevention of superimposed AKI or hospitalisations, as well as to address barriers to implementation.


Psychiatrists' awareness of partial and nonadherence to antipsychotic medication in schizophrenia: results from an Asia-Pacific survey.
Nonadherence is a well-known problem among schizophrenia patients, among whom relapse is fivefold more likely, adversely affecting health, employment, and social functioning. The Spanish Adherencia Terapéutica en la Esquizofrenia (ADHES) survey was developed to determine the scope and causes of medication nonadherence in schizophrenia. The 20-question ADHES survey was distributed to 19,370 psychiatrists in 13 Asia-Pacific countries in January-April 2012, to ascertain psychiatrists' perceptions of antipsychotic medication adherence levels among their schizophrenia patients, reasons for partial/nonadherence, their preferred methods of assessing adherence, and strategies to improve adherence. Responses are reported as mean and range across countries. Four thousand, six hundred sixty one psychiatrists (24% of recipients) completed the survey (highest contributors: People's Republic of China, 1854; India, 1616). Psychiatrists perceived that 56% (range, 30%-71%) of schizophrenia patients were non- or partially adherent to medication. Patients discontinue medication primarily due to lack of insight into their condition (mean, 37%; 1%-65%) and because patients consider medication unnecessary when feeling better (mean, 27%; 15%-68%). Over half of psychiatrists (mean, 55%; 42%-99%) assess medication adherence at every visit, almost exclusively (81%) by asking their patients, versus quantitative measures. One in three psychiatrists expressed their preference to switch to or add a long-acting antipsychotic to improve adherence (15%-82%). The substantial prevalence of partial/nonadherence to medication demonstrates that more proactive management of patients with schizophrenia is needed to improve adherence and thereby treatment outcomes. Registration of this study was not required.


Circular and circularly permuted forms of bovine pancreatic trypsin inhibitor.
Two novel forms of bovine pancreatic trypsin inhibitor have been prepared. The amino- and carboxyl-termini, which are in close proximity in the native conformation, were linked together in a peptide bond, thus generating a molecule with a circular backbone. The circular molecule was then cleaved between Lys15 and Ala16, to yield a linear molecule whose sequence is a circular permutation of that of bovine pancreatic trypsin inhibitor. Both of these modified forms could refold to the native conformation after being reduced, and promise to be interesting subjects for further folding experiments.


Immunoblastic lymphadenopathy with pulmonary infiltrates, hypocomplementemia and vasculitis. A hyperimmune syndrome.
A detailed description of the clinical and morphologic characteristics of four patients with immunoblastic lymphadenopathy, pulmonary infiltrates, hypocomplementemia and vasculitis is presented. Noteworthy in the patients described here is the clinical similarity to patients with collagen-vascular diseases and systemic drug reactions. The frequent occurrence of pulmonary symptoms, bilateral interstitial infiltrates and pleural effusions is emphasized. The morphology suggests stimulation of the immune system by antigenic agents, and the low complement levels and the presence of vasculitis suggest that circulating immune complexes may be present. Immunoblastic lymphadenopathy may represent a syndrome in which the adenopathy is but a nonspecific part of a systemic hyperimmune response to unknown antigens. The clinical course of the patients reviewed suggests that supportive treatment and corticosteroids constitute the safest therapeutic approach.


Isoflurane prevents transitions to tonic and burst firing modes in thalamic neurons.
We used patch-clamp whole-cell techniques to assess the effects of a volatile anaesthetic on thalamic firing modes in rat brain slices. Isoflurane application in clinical concentrations (0.5-2%) reversibly prevented voltage-dependent transitions to repetitive spike and burst firing modes in ventrobasal neurons. In voltage-clamp studies, isoflurane increased leak conductance, which shunted tonic and burst firing. Isoflurane also blocked the low-threshold Ca(2+)-current underlying the burst mode of firing, by increasing leak current and depressing membrane Ca(2+)-channel activity. We suggest that the mechanism of anaesthesia is distinct from sleep, although both states critically involve excitabilities of thalamic neurons.


Caveolin-1/-3: therapeutic targets for myocardial ischemia/reperfusion injury.
Myocardial ischemia/reperfusion (I/R) injury is a major cause of morbidity and mortality worldwide. Caveolae, caveolin-1 (Cav-1), and caveolin-3 (Cav-3) are essential for the protective effects of conditioning against myocardial I/R injury. Caveolins are membrane-bound scaffolding proteins that compartmentalize and modulate signal transduction. In this review, we introduce caveolae and caveolins and briefly describe the interactions of caveolins in the cardiovascular diseases. We also review the roles of Cav-1/-3 in protection against myocardial ischemia and I/R injury, and in conditioning. Finally, we suggest several potential research avenues that may be of interest to clinicians and basic scientists. The information included, herein, is potentially useful for the design of future studies and should advance the investigation of caveolins as therapeutic targets.


Tandemly repeated DNA sequences from Xenopus laevis. II. Dispersed clusters of a 388 base-pair repeating unit.
A repetitive DNA sequence family from Xenopus laevis that has an unusual genomic organization has been identified. It has been shown by blot-hybridization that this sequence occurs in clusters containing variable numbers of the 388 base-pair repeating unit. There are approximately 500 such clusters in the genome, and each cluster has common flanking sequences. The number of tandem 388 base-pair repeats per cluster ranges from one to at least 15, with a mean of seven. Homologous sequences were found in two related species, Xenopus borealis and Xenopus mulleri, where the size of the repeating units and their genomic arrangement are very similar to those in X. laevis. The complete nucleotide sequence of a cloned representative 388 base-pair repeating unit showed no short internal repeats and no long reading frames. By blot-hybridization, no evidence of transcripts of this sequence was found in total RNA from X. laevis liver, embryos or oocytes.


Risperidone in the Emergency Setting is Associated with More Hypotension in Elderly Patients.
Expert consensus panels have recommended risperidone as first-line treatment for agitation of psychiatric origin. However, there are few if any studies on this medication in the emergency setting. To assess the hemodynamic effects of risperidone in an emergency department (ED) setting, stratified by age. This is a structured chart review of all patients who received oral risperidone over a 6-year period in an ED setting, excluding patients who received this medication as a prescription refill. Vital signs were analyzed for this subset prior to and after medication administration, and changes in vital signs were stratified by age. The median dose of risperidone was less in patients aged > 65 years. However, the median drop in systolic blood pressure was larger in this age group compared with younger patients. Clinicians tend to be more cautious with dosing of risperidone to geriatric patients in the ED. Despite this, decreases in systolic blood pressure are larger and more frequent in this age group. When possible, clinicians should consider or attempt nonpharmacologic methods of agitation treatment prior to administering medications such as risperidone to elderly patients.


Statistical dependence between neighboring retinal ganglion cells in goldfish.
Simultaneous recordings were made of the spontaneous discharge of neighboring retinal ganglion cells in goldfish. Crosscorrelation analysis indicates two types of correlation. Neighboring units with like receptive field organization tend to discharge together while units having complementary receptive field organization tend not to discharge together.


Efficacy and safety of superselective trans-catheter arterial embolization of upper and lower gastrointestinal bleeding using N-butyl-2-cyanoacrylate.
To evaluate the efficacy and safety of superselective trans-catheter arterial embolization (TAE) with N-butyl-2-cyanoacrylate (NBCA) for patients with acute non-variceal upper and lower gastrointestinal (GI) bleeding. TAE using NBCA was performed in 21 patients (13 males, 8 females, mean age 60.9 years) with acute non-variceal upper (n = 15) and lower (n = 6) GI bleeding. TAE using other embolic agents was performed in 25 patients (13 males, 12 females, mean age 69.1 years) with acute non-variceal upper (n = 16) and lower (n = 9) GI bleeding. Technical success, clinical success, clinical failure, major and minor complications, bleeding-related 30-day mortality, and overall in-hospital mortality were analyzed. In 21 patients who underwent TAE with NBCA, the overall technical success was 100% (21/21) and overall clinical success was 72.2% (13/18). Uncontrolled bleeding, recurrent bleeding, and clinical failure were seen in 11.1% (2/18), 16.7% (3/18), and 27.8% (5/18) of cases, respectively. The minor complication rate was 16.7% (4/21) and no major complications occurred. Three patients showed ischemic damage in the treated lesion and one patient showed epigastric pain. The bleeding-related 30-day mortality and overall in-hospital mortality rates were 16.7% (3/18) and 28.6% (6/21), respectively. Two patients died of septic shock and one died of myocardial infarction within 30 days after TAE. In 25 patients who underwent TAE with other agents, the overall technical success was 100% (25/25), and the overall clinical success was 68.2% (15/22). The rates of uncontrolled bleeding, recurrent bleeding, clinical failure, bleeding-related 30-day mortality, and overall in-hospital mortality were 0.5% (1/22), 22.7% (5/22), 31.8% (7/22), 22.7% (5/22), and 32.0% (8/25), respectively. TAE with NBCA for acute non-variceal upper and lower GI bleeding proved to be a technically feasible, safe, and effective treatment modality. Thus, NBCA could be used as a primary embolic agent for controlling GI bleeding.


Cross-correlation between a carbonyl C' chemical shift anisotropy and a long-range dipolar C'HA coupling in proteins using symmetrical reconversion.
A new sequence is described to measure the cross-correlation rates between the chemical shift anisotropy of the carbonyl carbon-13 nucleus and the dipole-dipole interaction between this carbonyl and the alpha-proton in proteins. The sequence is based on the symmetrical reconversion principle and is insensitive to experimental errors and to violations of the secular approximation. The cross-correlation rate depends on the backbone angle psi. The advantages and limitations of the sequence are discussed.


Duchenne muscular dystrophy respiratory profiles from real world registry data.
Understanding real-world profiles from neuromuscular databases is helpful for optimizing clinical care and planning research studies. The Canadian Neuromuscular Disease Registry (CNDR) has respiratory data from a population of boys with Duchenne Muscular Dystrophy (DMD). To describe cross-sectional respiratory profiles from a national DMD real-world dataset. To explore the relationship between forced vital capacity percent predicted (FVC%) and disease severity parameters: scoliosis, ambulation and ventilation status. Descriptive statistics summarized the respiratory profiles. The CNDR registry enrolls and collects DMD clinic data from 36 Canadian centers. There were 414 participants enrolled. The age ranged from 2 to 36 years old. Pulmonary function test data were available for 323 participants. The use of ventilatory support was seen in a significant proportion (19.5%) of subjects by age 14-16 years and was used by the majority (69.2%) by age 20-22 years. FVC% declined at a rate of 3.19% per year with every 1-year increase in age. FVC% declined annually by 2.47% in nonambulatory participants versus by 0.96% in ambulatory participants. FVC% did not significantly change over age with the presence of scoliosis or use of ventilatory technology. The data from this large cohort are valuable for understanding real-world patterns of clinical care and disease progression. There is a significant association between the loss of ambulation and the rate of FVC% decline. Further longitudinal studies are needed to better understand the impact of disease parameters on pulmonary function decline and the need for ventilatory support.


Six-month outcomes for medical patients with major depressive disorders.
The courses of depressive disorders and factors affecting recovery rates among patients treated in primary medical care facilities have rarely been studied. The authors investigated the clinical status of 274 patients initially and six months after they sought care at one of three primary care sites. Through use of the highly structured Diagnostic Interview Schedule, they found that physicians assigned a depressive diagnosis to only a fourth of the patients so assessed by the structured interview. Nevertheless, the rates of persisting major depressive disorders at follow-up were found to be very similar for patients whom both the structured interview and the physician initially considered depressed and those so diagnosed by structured interview but not by the physician (25% and 31%, respectively). An analysis of factors associated with course of illness suggests that psychiatric status at the initial assessment and the number of assigned medical diagnoses rather than the physician's recognition and treatment of depression strongly predict continued affective disorder.


Natural cleanup of heavy fuel oil on rocks: an in situ experiment.
Changes in the chemical composition of a heavy fuel oil, Bunker C, exposed to the elements for 556 days in the vicinity of Brest Harbour (France, (48 degrees 18(') N, 4 degrees 32(') W)) have been studied. Samples with exposure to full or reflected sunlight, and in the dark, were analysed by thin layer chromatography and gas chromatography coupled with mass spectrometry and compared with the initial oil. Using hopane as a conserved internal standard, an average of more than 56% of the total hydrocarbon in the residual stranded oil had been removed in the 556 days. The results indicate that dissolution, biodegradation and photooxidation all play important roles in the weathering process, with their respective contributions depending on the exposure.


In silico microarray probe design for diagnosis of multiple pathogens.
With multiple strains of various pathogens being sequenced, it is necessary to develop high-throughput methods that can simultaneously process multiple bacterial or viral genomes to find common fingerprints as well as fingerprints that are unique to each individual genome. We present algorithmic enhancements to an existing single-genome pipeline that allows for efficient design of microarray probes common to groups of target genomes. The enhanced pipeline takes advantage of the similarities in the input genomes to narrow the search to short, nonredundant regions of the target genomes and, thereby, significantly reduces the computation time. The pipeline also computes a three-state hybridization matrix, which gives the expected hybridization of each probe with each target. Design of microarray probes for eight pathogenic Burkholderia genomes shows that the multiple-genome pipeline is nearly four-times faster than the single-genome pipeline for this application. The probes designed for these eight genomes were experimentally tested with one non-target and three target genomes. Hybridization experiments show that less than 10% of the designed probes cross hybridize with non-targets. Also, more than 65% of the probes designed to identify all Burkholderia mallei and B. pseudomallei strains successfully hybridize with a B. pseudomallei strain not used for probe design. The savings in runtime suggest that the enhanced pipeline can be used to design fingerprints for tens or even hundreds of related genomes in a single run. Hybridization results with an unsequenced B. pseudomallei strain indicate that the designed probes might be useful in identifying unsequenced strains of B. mallei and B. pseudomallei.


The role of study quality in aspartame and cancer epidemiology study reviews.
Toews et al. [1] and the World Health Organization (WHO) [2] reviewed observational epidemiology studies of non-sugar sweeteners (NSSs) and various health effects. The former used the Risk of Bias in Non-randomised Studies - of Interventions (ROBINS-I) tool and the latter used both the ROBINS-I tool and the Newcastle-Ottawa Scale to evaluate study quality. Both reviews concluded that there were no associations between NSS or aspartame consumption and cancer (except possibly between saccharin and bladder cancer) but indicated that the certainty of the evidence for all cancer types was "very low." While we agree with this conclusion, the support for the confidence in the evidence generally was not transparently documented, as the results of the study quality assessment were only provided in scores or ratings. An examination of illustrative case studies shows that some important aspects of study quality domains specific for NSSs generally or aspartame specifically (i.e., issues with the exposure and outcome assessments, the consideration of confounding/covariates, and selection bias) may have been overlooked or not given appropriate consideration, while other aspects that were less likely to have a large impact on overall study quality dominated the results in the two assessments. Our review of other studies published after the WHO [2] review further demonstrates this point. While this may not seem important given the overall lack of associations, it impacts the degree to which evidence supports a lack of effects as opposed to not being adequate to evaluate associations. In the future, aspartame and cancer outcome reviews should focus on those study quality domains that are most likely to impact the interpretation of results and discuss them in a transparent, systematic manner. If there is very low certainty in the evidence as a result of low study quality, reviewers should conclude the evidence is inadequate for making a causal determination.


Stationary phase retention and preliminary application of a spiral disk assembly designed for high-speed counter-current chromatography.
A spiral disk assembly composed of five single-channel units was designed for high-speed counter-current chromatography (HSCCC). The retention of different solvent systems ranging from moderately polar to polar organic-aqueous systems to aqueous two-phase systems (ATPs) was investigated under different elution modes. The results indicated that the spiral disk assembly can produce excellent retention of stationary phase for moderately polar organic-aqueous solvent systems, such as chloroform-methanol-water (4:3:2) and hexane-ethyl acetate-methanol-water (1:1:1:1) by pumping lower mobile phase from head (H) to tail (T), and upper mobile phase from tail (T) to head (H) even at a high flow-rate (8 mL/min, Sf>70%), regardless of whether the inlet is at the inner or outer terminal of the channel. This makes it possible for fast analysis of some small molecular compounds. This has been proved in the separation of mixtures of three flavones, including isorhamnetin, kaempferol, and quercetin. The spiral disk assembly can also provide satisfactory retention for polar to ATPS such as 1-butanol-acetic acid-water (4:1:5) (<3 mL/min, Sf>70%), 12.5% poly(ethylene glycol) (PEG) 1000-12.5% K2HPO4-75% water (< or =1 mL/min, Sf>70%) and 4% PEG 8000-5% Dextran T500-91% water (< or =0.5 mL/min, Sf>50%) by pumping lower mobile phase from inner terminal (I) to outer terminal (O), and upper mobile phase from outer terminal (O) to inner terminal (I) at a low flow-rate, while this is not possible with the multilayer coil column. Acceptable resolutions were achieved when it was used for the separation of peptides such as Leu-Tyr and Val-Tyr, and proteins including cytochrome c and myoglobin, lysozyme and myoglobin, and fresh chicken egg-white proteins.


Antiphospholipid antibodies and sub-clinical atherosclerosis in the Coronary Artery Risk Development in Young Adults (CARDIA) cohort.
Antiphospholipid antibodies (APA) have been associated with clinical cardiovascular disease, but it remains unclear whether APA are associated with sub-clinical atherosclerosis. This study examined the relationship between APA and sub-clinical atherosclerosis, measured as coronary artery calcification (CAC), in participants from the prospective Coronary Artery Risk Development in Young Adults (CARDIA) Study. 2,203 black and white participants with sera available from the CARDIA year 7 examination and CAC measured by computed tomography at years 15 or 20 were selected. Anti-β2-glycoprotein I (anti-β2-GPI) immunoglobulin (Ig) M, IgG, and IgA were positive in 7.0, 1.4, and 1.8 % of participants, respectively; anti-cardiolipin (aCL) IgM and IgG were positive in 1.5 and 1.0 %, respectively. 9.5 % of participants had CAC score >0 at year 15. Anti-β2-GPI IgM, IgG, IgA, and aCL IgG positivity were associated with CAC >0 at year 15 after adjustment for traditional cardiovascular risk factors; [odds ratios (95 % confidence intervals) were 1.7 (1.0, 3.1), 6.4 (2.4, 16.8), 5.6 (2.3, 13.2), and 5.1 (1.4, 18.6), respectively]. Anti-β2-GPI IgG was associated with year 20 CAC >0, and anti-β2-GPI IgA and aCL IgG were marginally associated. These findings indicate that APA positivity during young adulthood is a risk factor for subsequent sub-clinical atherosclerosis and might play a role in the pathogenesis of atherosclerosis


A disulfide-free single-domain V(L) intrabody with blocking activity towards huntingtin reveals a novel mode of epitope recognition.
We present the crystal structure and biophysical characterization of a human V(L) [variable domain immunoglobulin (Ig) light chain] single-domain intrabody that binds to the huntingtin (Htt) protein and has been engineered for antigen recognition in the absence of its intradomain disulfide bond, otherwise conserved in the Ig fold. Analytical ultracentrifugation demonstrated that the αHtt-V(L) 12.3 domain is a stable monomer under physiological conditions even at concentrations >20 μM. Using peptide SPOT arrays, we identified the minimal binding epitope to be EKLMKAFESLKSFQ, comprising the N-terminal residues 5-18 of Htt and including the first residue of the poly-Gln stretch. X-ray structural analysis of αHtt-V(L) both as apo protein and in the presence of the epitope peptide revealed several interesting insights: first, the role of mutations acquired during the combinatorial selection process of the αHtt-V(L) 12.3 domain-initially starting from a single-chain Fv fragment-that are responsible for its stability as an individually soluble Ig domain, also lacking the disulfide bridge, and second, a previously unknown mode of antigen recognition, revealing a novel paratope. The Htt epitope peptide adopts a purely α-helical structure in the complex with αHtt-V(L) and is bound at the base of the complementarity-determining regions (CDRs) at the concave β-sheet that normally gives rise to the interface between the V(L) domain and its paired V(H) (variable domain Ig heavy chain) domain, while only few interactions with CDR-L1 and CDR-L3 are formed. Notably, this noncanonical mode of antigen binding may occur more widely in the area of in vitro selected antibody fragments, including other Ig-like scaffolds, possibly even if a V(H) domain is present.


The induction of DNA strand breaks at specific sites by N-nitroso-N-ethylurea depends on the phases of the cell cycle.
Synchronized root meristems of Pisum sativum were treated at each phase of the cell cycle with 6.25 mM N-nitroso-N-ethylurea. DNA extracted from treated cells and run in agarose gel electrophoresis exhibits a series of discrete fragments with length below 2500 bp and a significant number of unspecific single-stranded breaks (or alkali-labile sites). Experiments with micrococcal nuclease indicated that the nucleosomal organization of the chromatin is not responsible for the generation of the discrete fragments: it seems that their appearance is associated with a preferable attack of the mutagen at specific sites, characteristic for the plant genome. Moreover, a cell cycle dependent release of the discrete fragments was found with maximum at G1-S and minimum at mitosis. The model experiments designed to clarify this observation suggest that it might be determined from the cell cycle dependent fluctuation in the accessibility of the chromatin DNA and/or the process of excision-repair.


Impact of spatial resolution on results of esophageal high-resolution manometry.
The Chicago classification for esophageal motility disorders was designed for a 36-channel manometry system with sensors spaced at 1 cm. However, many motility laboratories outside the USA use catheters with a lower resolution in the segments outside the esophagogastric junction. Our aim was to investigate the effect of spatial resolution on the Chicago metrics and diagnosis. In 20 healthy volunteers and 47 patients with upper gastrointestinal symptoms, high-resolution manometric studies of the esophagus were retrospectively reanalyzed using the original 1-cm spacing in the segments outside the 7-cm esophagogastric junction segment, and again after manually increasing the spacing between sensors to 2, 3, and 4 cm (above the lower esophageal sphincter region). Measurements were analyzed in random order and the investigator was blinded to the outcome of the analyses performed in another resolution of the same patient. Intra-class correlation coefficients (ICC) and Kappa values were determined. There was a very strong correlation between the 1-cm and 2-cm analysis for all Chicago metrics studied in healthy volunteers (ICCs: distal contractile integral 0.998; contractile front velocity (CFV) 0.964; distal latency 0.919; peristaltic break size 0.941). The 2-cm spacing analysis also correlated very well with the 1-cm analysis for the different Chicago diagnoses obtained in the patients (Kappa values ranging from 0.665 to 1.000). When the sensor spacing was increased to 3 and 4 cm, the correlation was reduced to moderate for the Chicago metrics, especially for break size and CFV of peristalsis. The Chicago classification for esophageal motility disorders is still valid and the same normal values can be used when catheters with a slightly lower resolution are used (i.e., 2-cm vs 1-cm spacing). For larger sensor intervals, the classification and the normal values will need to be adjusted.


Interaction of substance P with dispersed pancreatic acinar cells from the guinea pig. Relationships between structure, binding and biological activity.
Binding of 125I-[Tyr8]-SP to isolated pancreatic acinar cells was inhibited in a concentration-dependent way by SP, [Tyr8]-SP and longer C-terminal fragments of SP. SP6-11 was the shortest sequence to bind significantly to SP-receptors as well as to stimulate amylase release from dispersed pancreatic acini. SP7-11 and shorter fragments did not inhibit binding of 125I-[Tyr8]-SP and did not stimulate secretion of amylase significantly. SP augmented the stimulatory effect of cholecystokinin on amylase release. Two SP-antagonists, [D-Pro2, D-Trp7,9]-SP and [D-Pro2,4, D-Lys3, D-Gln5,6, D-Trp7,9]-SP inhibited binding of 125I-[Tyr8]-SP in a concentration dependent manner and tended at a high concentration to reduce release of amylase evoked by submaximal concentrations of SP.


Abstinence in late adolescence--antecedents to and covariates of a sober lifestyle and its consequences.
The purpose of this study was first to compare 18-19-year-old male abstainers with alcohol consumers, and especially light consumers, regarding degree of sociability as indicated by their (in)security in the company of others, their number of close friends, intimate conversations with friends and their popularity in school. Secondly, we analysed the importance of antecedents to and covariates of abstinence. In addition, the significant antecedents and covariates gave us information as to abstinence patterns. The study was based on a survey of all Swedish males, 18-19 years old, conscripted for military service in 1969-70. Data had been collected by means of questionnaires and psychological interviews, giving measures of each respondent's social background, psychiatric/psychological and psychosomatic health status, substance use, deviant behaviour and degree of sociability. Poor sociability was more common among the abstainers than among all the other categories of drinkers, including the light consumers. The conscripts' social background, and especially their fathers' drinking habits, had the strongest effects in explaining abstinence. Sixty-two per cent of all abstainers had non-drinking fathers, compared to 28% of the light consumers. As to the majority of abstainers, this indicates a link between the social background of temperance and their own reported abstinence. Their poor sociability could be a consequence of abstaining at a young age when abstinence is uncommon. Those who abstained despite a drinking father showed a worsening psychological status, suggesting a link between psychologically impaired health, poor sociability and abstinence. Though the abstainers were the least sociable, the difference between the abstainers, the light consumers and the moderate consumers in other categories were generally small.(ABSTRACT TRUNCATED AT 250 WORDS)


Bayesian sparse reduced rank multivariate regression.
Many modern statistical problems can be cast in the framework of multivariate regression, where the main task is to make statistical inference for a possibly sparse and low-rank coefficient matrix. The low-rank structure in the coefficient matrix is of intrinsic multivariate nature, which, when combined with sparsity, can further lift dimension reduction, conduct variable selection, and facilitate model interpretation. Using a Bayesian approach, we develop a unified sparse and low-rank multivariate regression method to both estimate the coefficient matrix and obtain its credible region for making inference. The newly developed sparse and low-rank prior for the coefficient matrix enables rank reduction, predictor selection and response selection simultaneously. We utilize the marginal likelihood to determine the regularization hyperparameter, so our method maximizes its posterior probability given the data. For theoretical aspect, the posterior consistency is established to discuss an asymptotic behavior of the proposed method. The efficacy of the proposed approach is demonstrated via simulation studies and a real application on yeast cell cycle data.


A novel method for the biological evaluation of protein quality using plasma somatomedin-C (insulin-like growth factor-1) measurement.
The objective of the present study was to investigate the validity of plasma somatomedin-C insulin-like Growth Factor-1 (IGF-1) measurement for the evaluation of protein quality. Six sources of protein were evaluated using the conventional methods: protein efficiency ratio (PER), net protein ratio (NPR), relative protein value (RPV), biological value (BV) and protein digestion coefficient (D). Plasma IGF-1 level was also measured on the third day of the feeding period. Plasma levels of IGF-1 ranked the protein sources in the same order as other conventional methods. In addition, values of IGF-1 correlated significantly with the values of PER, NPR, RPV and BV. Protein digestion values correlated poorly with the other methods being used. We conclude that the measurement of plasma IGF-1 is a valid, reliable, easy and rapid method for protein quality evaluation to be used by the food industry and compliance regulatory agencies. Furthermore, this method could be employed for the evaluation of protein quality in infant formulas and for pre-clinical evaluation of enteral formulas.


Human Keratinocytes Use Sphingosine 1-Phosphate and its Receptors to Communicate Staphylococcus aureus Invasion and Activate Host Defense.
Sphingosine 1-phosphate (S1P) is a bioactive lipid mediator generated when a cell membrane or its components are damaged by various factors. S1P regulates diverse cell activities via S1P receptors (S1PRs). Keratinocytes express S1PR1-5. Although it is known that S1PRs control keratinocyte differentiation, apoptosis, and wound healing, S1PR functions in keratinocyte infections have not been fully elucidated. We propose that the S1P-S1PR axis in keratinocytes works as a biosensor for bacterial invasion. Indeed, in human impetigo infection, we found high epidermal expression of S1PR1 and S1PR2 in the skin. Furthermore, in normal human epidermal keratinocytes in vitro, treatment with Staphylococcus aureus bacterial supernatant not only induced S1P production but also increased the transcription of S1PR2, confirming our in vivo observation, as well as increased the levels of TNFA, IL36G, IL6, and IL8 mRNAs. However, direct treatment of normal human epidermal keratinocytes with S1P increased the expressions of IL36G, TNFA, and IL8, but not IL6. In both S1P- and S. aureus bacterial supernatant-treated normal human epidermal keratinocytes, S1PR1 knockdown reduced IL36G, TNFA, and IL8 transcription, and the S1PR2 antagonist JTE013 blocked the secretion of these cytokines. Overall, we have proven that during infections, keratinocytes communicate damage by using S1P release and tight control of S1PR1 and 2.


Fasting glucose and postprandial glycemia: which is the best target for improving outcomes? The Apollo and 4-T Trials.
Two studies, the Apollo and 4-T Trials, were conducted in order to determine which insulin regimen (basal or prandial) is the most efficient for the treatment of insulin-requiring type 2 diabetic patients. Both trials compared treatments using prandial insulins (aspart or lispro) three times daily with more classical insulin strategies using basal insulin given once daily (glargine or detemir) or twice daily if required (detemir in the 4-T Study). Both studies showed that a therapeutic regimen involving prandial insulin resulted in equal (Apollo Study) or greater (4-T Study) reductions in patients' HbA(1c) levels than basal insulin regimens. However, the prandial insulin strategies were accompanied by higher risks of hypoglycemia and greater weight gain. As a consequence, the investigators of the two studies concluded that basal insulin once daily provides a simple and effective option with less adverse effects than prandial insulin three times a day. Such conclusions are certainly important for guiding strategies in the vast majority of type 2 diabetic patients who require an add-on insulin therapy. However, the authors' opinion is that the choice between either basal or prandial insulin alone and combined insulin regimens with basal and prandial insulin should be tailored according to the patient's clinical status by paying more attention to the respective contributions of basal and prandial hyperglycemia to their overall hyperglycemia. This recommendation seems to be particularly important when insulin treatment is initiated in patients exhibiting HbA(1c) levels between 7.0 and 8.0%.


In-process monitoring of a tissue-engineered oral mucosa fabricated on a micropatterned collagen scaffold: use of optical coherence tomography for quality control.
We previously reported a novel technique for fabricating dermo-epidermal junction (DEJ)-like micropatterned collagen scaffolds to manufacture an ex vivo produced oral mucosa equivalent (EVPOME) for clinical translation; however, more biomimetic micropatterns are required to promote oral keratinocyte-based tissue engineering/regenerative medicine. In addition, in-process monitoring for quality control of tissue-engineered products is key to successful clinical outcomes. However, evaluating three-dimensional tissue-engineered constructs such as EVPOME is challenging. This study aimed to update our technique to fabricate a more biomimetic DEJ structure of oral mucosa and to investigate the efficacy of optical coherence tomography (OCT) in combination with deep learning for non-invasive EVPOME monitoring. A picosecond laser-textured microstructure mimicking DEJ on stainless steel was used as a negative mould to fabricate the micropatterned collagen scaffold. During EVPOME manufacturing, OCT was applied twice to monitor the EVPOME and evaluate its epithelial thickness. Our moulding system resulted in successful micropattern replication on the curved collagen scaffold. OCT imaging visualised the epithelial layer and the underlying micropatterned scaffold in EVPOME, enabling to non-invasively detect specific defects not found before the histological examination. Additionally, a gradual increase in epithelial thickness was observed over time. These findings demonstrate the feasibility of using a stainless-steel negative mould to create a more biomimetic micropattern on collagen scaffolds and the potential of OCT imaging for quality control in oral keratinocyte-based tissue engineering/regenerative medicine.


Plaque formation by a host range mutant of vaccinia virus in non-permissive cells co-infected with Yaba virus.
A host-dependent conditional lethal mutant of vaccinia virus strain DIs was rescued to form plaques in a non-permissive cell line JINET co-infected with Yaba virus, a poxvirus serologically distant from the rescued virus. The efficiency of plaque formation under optimal conditions in this system was comparable to that in Vero cells which were permissive for the mutant. The plaque number of the mutant in JINET cells was influenced greatly by the multiplicity of Yaba virus, the interval between inoculations with DIs and Yaba virus and the maintenance medium for pre-infection of cells with Yaba virus. The plaque formation by DIs in JINET cells was suppressed or inhibited when the duration of pre-infection with high multiplicities of Yaba virus was prolonged. Thus, Yaba virus possessed both rescuing and inhibitory activities on DIs and the plaque number of DIs in doubly infected cells was thought to be determined by the balance between the two activities. Ultraviolet light-inactivated Yaba virus retained rescuing activity on DIs in JINET cells.


The diagnostic significance of antibody specificity indices in multiple sclerosis and herpes virus induced diseases of the nervous system.
The antibody specificity index (ASI) indicates the cerebrospinal fluid (CSF)/serum difference of antibody amounts per weight unit IgG (normal less than 1.5). It has proven to be the most sensitive inflammation parameter in CSF analysis so far, more sensitive than the Western blot, the "oligoclonal" response, and the empirical differentiation of CSF immunoglobulins. By this diagnostic criterion, several benign viral meningitis cases were found to be caused by the varicella/zoster virus. The diagnostic relevance of local zoster antibody synthesis was greatest in ganglionitis cases, e.g., in zoster oticus sine herpete (facial paresis) and acute radicular syndromes of the elderly. The diagnostic significance of the local immune response against measles, rubella, and zoster antigens (MRZ response) was ascertained further. Together with oligoclonal gamma-globulin fractionation, there is now only 1 out of 100 multiple sclerosis (MS) patients left who has been found to have a normal CSF.


Coexistence of three rare congenital heart defects in a single patient: a unique case with important embryologic implications.
The unique anatomy, physiology, and surgical repair of a patient with features of three rare congenital heart defects (total anomalous pulmonary venous return to the coronary sinus, cor triatriatum, and unroofed coronary sinus with persistent left superior vena cava to the left atrium) is described. Analysis of this case suggests that these three conditions are linked in that they all may result as a consequence of a defect in the same embryologic process (i.e., incorporation of the pulmonary venous confluence into the left atrium).


Intranasal drug-induced fungal rhinopharyngitis.
Intranasal drug abuse has long been recognized as an etiology of sinonasal pathology. However, the intranasal use of prescription and nonprescription drugs has surpassed the use of illicit drugs, and the pattern of presentation and required therapeutic intervention appears to be different. We report on our experience with these patients, along with a successful treatment algorithm for this disease process. Retrospective chart review of 9 consecutive patients who presented with rhinopharyngitis and a history of intranasal opioid and/or acetaminophen abuse, from 2007 to 2010, at a tertiary referral center. Nine patients were found to have abused intranasal hydrocodone/acetaminophen, oxycodone/aceta-minophen, or acetaminophen and were diagnosed with rhinopharyngitis. Sinonasal pain and odynophagia were the most common chief complaints and 8 of 9 patients reported previous antibiotic failures. On endoscopy, all patients exhibited a thick, white, exudative process involving the nasal septum and lateral nasal mucosa. Five of 9 exhibited large septal perforations. Seven of 9 exhibited white, exudative pharyngitis. Seven of 9 patients had identifiable fungal organisms on culture, including 5 with species of Candida and 3 with Aspergillus. Two patients grew Staphylococcus aureus. Five patients were compliant with follow up. All 5 showed significant improvement in symptoms and examination, following treatment with oral and topical antifungal therapy and nasal irrigations. Intranasal opioid and acetaminophen abuse is often associated with the development of fungal rhinopharyngitis. Recognizing that this process is primarily fungal in origin is paramount to successful treatment, as most patients respond well to antifungal therapy when compliant with treatment.


The nucleoprotein gene of Ebola virus: cloning, sequencing, and in vitro expression.
Genomic and messenger RNAs of a Zaire strain of Ebola virus were cloned, and inserts specific for the nucleoprotein gene were isolated and sequenced. The nucleoprotein gene is located proximal to the 3' end of the genome and is preceeded by a putative leader sequence. The gene begins with the transcriptional start site sequence 3'-UACUCCUUCUAAUU..., and ends with the polyadenylation site sequence 3'-... UAAUUCUUUUUU. The predicted coding region is 2217 bases in length and encodes a protein that contains 739 amino acids, with a calculated molecular weight of 83.3 kDa. The protein has an approximate net charge of -30 and can be divided into a hydrophobic N-terminal half and a hydrophilic and highly acidic C-terminal half. An in vitro transcript, generated from plasmid DNA containing the entire coding region, directs the synthesis of authentic nucleoprotein in a rabbit reticulocyte lysate system. The genomic organization and transcriptional signals of Ebola are similar to those of other nonsegmented, negative-strand RNA viruses, but nucleic acid or amino acid sequence comparisons indicate a lack of similarity.


Cochlear outer hair cell electromotility can provide force for both low and high intensity distortion product otoacoustic emissions.
It is generally believed that the force for the otoacoustic emission (OAE) generation is provided by a mechanism of electromotility, observed in isolated cochlear outer hair cells (OHCs). OHC electromotility is resistant to several ototoxic reagents, it does not depend on ATP hydrolysis, but it can be blocked by specific sulfhydryl reagents: p-chloromercuriphenylsulfonic acid (pCMPS) and p-hydroxymercuriphenylsulfonic acid (pHMPS). We have used these reagents to test whether they also affect OAE. Application of pCMPS and pHMPS on the round window membrane of anesthetized guinea pigs produced a dose-dependent inhibition of the cubic (2F1-F2) distortion product OAE (DPOAE). The inhibition developed progressively from high to low frequencies, reflecting the diffusion of the drugs through the cochlear compartment. The effect of pCMPS and pHMPS was different from the effects of furosemide and lethal anoxia, which impair cochlear function but do not block OHC electromotility. pHMPS suppressed DPOAE completely at all sound intensities tested (45-80 dB SPL), whereas furosemide or lethal anoxia caused DPOAE to disappear at low-level stimulation (45-60 dB SPL) only. Our results suggest that the OHC electromotility might provide the force for DPOAE generation not only at low, but also at high stimulus intensities.


Higher formant normalization results from auditory integration of F2 and F3.
Two experiments were conducted to test the hypothesis that higher formant normalization results from the auditory integration of F2 and F3 when they are within 3 Bark of each other. In the first experiment, Formants 3-5 were manipulated in both a "hid"-"head" continuum (in which F2 and F3 are within 3 Bark of each other) and a "hood"-"HUD" continuum (in which F2 and F3 are not within 3 Bark of each other). It was found that there was a shift in identification consistent with the higher formant normalization effect only in the "hid"-"head" continuum. In the second experiment, F3 alone was manipulated in a "hood"-"HUD" continuum. The amplitude of F3 in this continuum was increased (as compared with the F3 in the "hood"-"HUD" continuum used in Experiment 1) and a pretest indicated that the shift in F3 could be detected. As in the first experiment, there was no shift in identification associated with shifting F3 frequency in a back-vowel continuum. The results of these experiments are not consistent with an explanation of higher formant normalization in which hearers adjust an internal vowel space in response to higher formant information; rather, the present findings indicate that higher formant normalization results from auditory integration of F2 and F3.


Radiotherapy of lymphomas.
Radiotherapy for Hodgkin lymphomas has evolved a lot over time, but still plays an important role, almost always in addition to chemotherapy, for the management of the early stages. The major objective is to preserve the quality of life of patients who will be cured from this disease in the vast majority of cases. Also, the personalization of the indications for the purpose of de-escalating toxicity is very refined and is essentially based on the pre- and pertherapeutic assessment by FDG-PET. The indications for radiotherapy are more limited for non-Hodgkin lymphomas, but the same principles are found, regardless of the histological type. We present the update of the recommendations of the French society of oncological radiotherapy for radiotherapy of lymphomas, which remains a very evolving field in terms of therapeutic strategy and evaluation.


Establishing cell polarity in development.
Polarity is a common feature of many different cell types, including the Caenorhabditis elegans zygote, the Drosophila oocyte and mammalian epithelial cells. The initial establishment of cell polarity depends on asymmetric cues that lead to reorganization of the cytoskeleton and polarized localization of several cortical proteins that act downstream of the polarization cues. The past year revealed that homologs of the C. elegans par (partitioning defective) genes are also essential for establishing polarity in Drosophila and vertebrate cells. There is growing evidence that the proteins encoded by these genes interact with key regulators of both the actin and the microtubule cytoskeletons.


When Cancer and Cardiovascular Disease Intersect: The Challenge and the Opportunity of Cardio-Oncology.
Cancer and cardiovascular disease (CVD) commonly coexist, with increasing evidence that long-term cancer survivors are more likely to die from CVD than the general population. Effective management of CVD and its risk factors requires identification of patients at increased risk who may benefit from early intervention and their appropriate monitoring across the disease trajectory. Improving outcomes requires new models of multidisciplinary cancer care supported by care pathways. Such pathways require a clear delineation of the roles and responsibilities of all team members and provision of appropriate enablers for their delivery. These include accessible point-of-care tools/risk calculators, patient resources, and the provision of tailored training opportunities for health care providers.


Use of Sep-Pak cartridges for on-line preparative high-performance liquid chromatography.
Sep-Pak C18 cartridges have found wide application to the extraction, concentration, and fractionation of peptides. Their use has hitherto been limited to stepwise loading, washing, and eluting procedures. By the use of commercially available plastic column end-fittings it was possible to connect steel tubing to both ends of the cartridges and allow low-pressure operation in a high-performance liquid chromatography (HPLC) instrument. The cartridge was then able to function as a disposable, reversed-phase (RP) preparative column. Operation in a gradient mode gave several advantages over step-wise elution, including the direct UV absorption monitoring of the eluant. Gradient conditions could be optimised for sample recovery. The cartridge was compared with standard analytical RP-HPLC columns for the separation of both proteins and peptides. A test protein gave a single peak with good recovery. Experiments with peptide mixtures showed that the cartridge could resolve some components sufficiently to allow fractions of pure peptides to be collected.


Tests for 'detrusor instability' in women. These mainly measure the urethral resistance created by pelvic floor contraction acting against a premature activation of the micturition reflex.
The principal aim of this study was to analyse the simultaneous pressure readings derived from bladder and urethra during a handwashing test. A total of 163 patients with urinary incontinence were studied. It was demonstrated that contraction of the pelvic floor stretches the vagina. In many patients, this appeared to inhibit the micturition reflex, possibly by supporting the nerve endings at bladder neck, thereby inhibiting their premature activation. There was also evidence that this stretching activated reflex contraction of the urethra, the vagina-urethral (coital) reflex, thereby increasing the urethral resistance. On further analysis, it was concluded that urodynamic testing does not detect 'detrusor instability' per se. It detects the urethra's attempts at closure.


The risk of gallbladder cancer from polyps in a large multiethnic series.
The aim of this study is assess whether patients with Indian ethnic background are at an increased risk of developing gallbladder cancer (GBC) if they have been diagnosed with ultrasonic abnormalities of the gallbladder. Between January 1998 and July 2006, 137,655 abdominal ultrasound examinations were performed in Leeds Teaching Hospitals NHS Trust. After the exclusion of repeat scans and those performed for renal or pelvic disease, 71,431 reports were included in this analysis. Patients in whom the diagnosis of GBC has been made without histology have been identified from the database of Northern and Yorkshire Cancer Registry and the presence of GBC was correlated with ultrasonic gallbladder abnormalities. Gallbladder polyps (GBP) were detected in 3.3% of patients and these were larger than 10 mm in 0.1% of the cases. Age above 60 years, Indian ethnic background, single GBP larger than 10mm, the presence of gallstones, severe gallbladder wall thickening and irregular thickening were independently associated with the higher odds of developing GBC. The prevalence of malignancy in those with GBP was significantly higher among patients with Indian ethnic background compared to Caucasian patients, 5.5% versus 0.08%, p<0.001. The presence of GBP, irrelevant of size, amongst patients of Indian ethnic decent, is an indication for further investigation and/or cholecystectomy.


Modelling compound action potentials of peripheral nerves in situ. II. A study of the influence of temperature.
Compound nerve action potentials of sural nerves of healthy volunteers have been measured at different temperatures. In the range from 21 to 39 degrees C a linear temperature dependence of the propagation velocity, related to the fastest fibres, is found (slope 1.9 m/sec/degree C). The causes of amplitude and shape alterations in the compounds action potentials (CAPs) with changing temperature are analysed with the help of a model. It appears that the changes in the measured CAPs are well explained by known temperature effects on the single fibre level. Both the considerable increase in CAP duration after cooling and the relative constancy of the CAP amplitude were reproduced by the model. Close fits to the actually measured CAPs can be obtained when some crucial parameter values are optimized. The results of this procedure are presented.


Increased epidermal filaggrin in chronic idiopathic urticaria is associated with severity of urticaria.
Chronic idiopathic urticaria (CIU) and atopic dermatitis (AD) are common allergic skin diseases associated with severe pruritus. AD skin is characterized by filaggrin deficiency, but it has not been studied in CIU. To compare the expression of filaggrin in skin from patients with CIU, patients with AD, and normal controls and to investigate whether altered filaggrin expression is associated with CIU severity. Skin biopsies were obtained from 16 patients with CIU, 11 patients with AD, and 14 normal controls. Filaggrin expression was evaluated using real-time reverse transcriptase polymerase chain reaction and immunostaining. Urticaria activity score, transepidermal water loss, and skin pH were measured. FLG gene expression was significantly greater in lesional CIU skin compared with lesional AD skin (P < .01). The staining intensity of filaggrin was significantly increased in lesional CIU skin compared with skin from normal controls (P < .01) and lesional AD skin (P < .001). A significant correlation was observed between filaggrin staining intensity and urticaria activity score in patients with CIU (r = 0.538, P < .05). Transepidermal water loss was significantly increased in lesional skin of patients with AD compared with skin from normal controls (P < .01) and lesional skin from patients with CIU (P < .01). Skin pH was significantly decreased in lesional skin from patients with CIU compared with skin from normal controls (P < .01) and patients with AD (P < .001). Filaggrin is overexpressed in lesional CIU skin, and increased filaggrin expression is positively correlated with urticaria severity in CIU. Altered filaggrin expression has physiologic effects on transepidermal water loss and pH in the skin of patients with CIU, suggesting increased barrier function compared with skin from patients with AD.


A simple bridging flocculation assay for rapid, sensitive and stringent detection of gene specific DNA methylation.
The challenge of bringing DNA methylation biomarkers into clinic is the lack of simple methodologies as most current assays have been developed for research purposes. To address the limitations of current methods, we describe herein a novel methyl-protein domain (MBD) enrichment protocol for simple yet rapid and highly stringent selection of highly methylated DNA from limiting input samples. We then coupled this with a DNA-mediated flocculation assay for rapid and low cost naked-eye binary evaluation of highly methylated genes in cell line and blood DNA. The low resource requirements of our method may enable widespread adoption of DNA methylation-based diagnostics in clinic and may be useful for small-scale research.


MicroRNA-145-5p regulates fibrotic features of recessive dystrophic epidermolysis bullosa skin fibroblasts.
Recessive dystrophic epidermolysis bullosa (RDEB) is a skin fragility disorder caused by mutations in the COL7A1 gene encoding type VII collagen, a cutaneous basement membrane component essential for epidermal-dermal adhesion. Hallmarks of the disease are unremitting blistering and chronic wounds with severe inflammation and fibrosis. MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression also implicated in fibrotic processes. However, the role of miRNAs in RDEB fibrosis is almost unexplored. Our study aimed to identify miRNAs deregulated in primary RDEB skin fibroblasts (RDEBFs) and to characterize their function in RDEB fibrosis. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to screen RDEBFs for expression levels of a group of miRNAs deregulated in hypertrophic scars and keloids, pathological conditions with abnormal wound healing and fibrosis. Contractility, proliferation and migration rate were evaluated by different in vitro assays in RDEBFs transfected with a miR-145-5p inhibitor. Expression levels of fibrotic markers and miR-145-5p targets were measured using qRT-PCR and western blot. The miR-143/145 cluster was upregulated in RDEBFs compared with fibroblasts from healthy subjects. RDEBFs transfected with a miR-145-5p inhibitor showed attenuated fibrotic traits of contraction, proliferation and migration, accompanied by reduced expression of the contractile proteins α-smooth muscle actin and transgelin. These effects were associated with upregulation of Krüppel-like factor 4 transcriptional repressor and downregulation of Jagged1, a known inducer of fibrosis. Our results highlight the profibrotic role of miR-145-5p and its regulatory networks in RDEB, shedding light on novel disease pathomechanisms and targets for future therapeutic approaches. What's already known about this topic? Recessive dystrophic epidermolysis bullosa (RDEB) is a highly disabling genetic skin disease caused by mutations in the collagen VII gene and characterized by unremitting blistering and defective wound healing, leading to inflammation and fibrosis. MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression in health and disease, and their deregulation has been implicated in fibrotic skin conditions. To date, only miR-29 has been associated with injury-driven fibrosis in RDEB. What does this study add? In patients with RDEB, miR-145-5p is overexpressed in RDEB skin fibroblasts (RDEBFs), where it plays a profibrotic role, as its inhibition reduces RDEBF fibrotic traits (contraction, proliferation and migration). miR-145-5p inhibition in RDEBFs determines the reduction of contractile markers α-smooth muscle actin and transgelin through upregulation of Krüppel-like factor 4, a transcriptional repressor of contractile proteins, and downregulation of Jagged1 (JAG1), an inducer of fibrosis. What is the translational message? Our findings expand the knowledge on miRNA-driven pathomechanisms implicated in RDEB fibrosis. miR-145-5p and its targets (e.g. JAG1) could represent relevant molecules for the development of novel therapeutic strategies to counteract fibrosis progression in patients with RDEB.