Third-hand Smoke: Impact on Hemostasis and Thrombogenesis.
Cigarette smoking is a major risk factor for acute coronary thrombosis. In fact, both active/first-hand smoke and passive/second-hand smoke exposure are known to increase the risk of coronary thrombosis. Although recently a new risk has been identified and termed third-hand smoke (THS), which is the residual tobacco smoke contaminant that remains after a cigarette is extinguished, it remains to be determined whether it can also enhance the risk of thrombogenesis, much like first-hand smoke and second-hand smoke. Therefore, the present studies investigated the impact of THS exposure in the context of platelet biology and related disease states. It was found that THS-exposed mice exhibited an enhanced platelet aggregation and secretion responses as well as enhanced integrin GPIIb-IIIa activation. Furthermore, it was found that THS exposure shortens the tail bleeding time and the occlusion time in a model of thrombosis. Thus, our data demonstrate for the first time (at least in mice) that THS exposure increases the risk of thrombosis-based disease states, which is attributed, at least in part, to their hyperactive platelets.


Topographical organization of the N-terminal segment of lung pulmonary surfactant protein B (SP-B(1-25)) in phospholipid bilayers.
The location and depth of each residue of lung pulmonary surfactant protein B (SP-B(1-25)) in a phospholipid bilayer (PB) was determined by fluorescence quenching using synthesized single-residue-substituted peptides that were reconstituted into 1,2-dipalmitoyl phosphatidylcholine (DPPC)-enriched liposomes. The single-residue substitutions in peptides were either aspartate or tryptophan. The aspartate was subsequently labeled with the N-cyclohexyl-N'-(4-(dimethylamino)naphthyl)carbodiimide (NCD-4) fluorophore, whereas tryptophan is autofluorescent. Spin-labeled compounds, 5-doxylstearic acid (5-DSA), 7-doxylstearic acid (7-DSA), 12-doxylstearic acid (12-DSA), 4-(N,N-dimethyl-N-hexadecyl)ammonium-2,2,6,6-tetramethylpiperidine-1-oxyl iodide (CAT-16), and 4-trimethylammonium-2,2,6,6-tetramethylpiperidine-1-oxy iodide (CAT-1), were used in the quenching experiments. The effective quenching order is determined by the accessibility of the quencher to a fluorescent group on the peptide. The order of quenching efficiency provides information about the relative locations of individual residues in the PB. Our data indicate that residues Phe1-Pro6 are located at the surface of PB, residues Tyr7-Trp9 are embedded in PB, and residues Leu10-Ile22 are involved in an amphipathic alpha-helix with its axis parallel to the surface of PB; residues Pro23-Gly25 reside at the surface. The effects of intermolecular disulfide bond formation in the SP-B(1-25) dimer were also investigated. The experiments suggest that the SP-B helix A has to rotate at an angle to form a disulfide bond with the neighboring cysteine, which makes the hydrophobic sides of the amphipathic helices face each other, thus forming a hydrophobic domain. The detailed topographical mapping of SP-B(1-25) and its dimer in PB provides new insights into the conformational organization of the lung pulmonary surfactant proteins in the environment that mimics the native state. The environment-specific conformational flexibility of the hydrophobic domain created by SP-B folding may explain the key functional properties of SP-B including their impact on phospholipid transport between the lipid phases and in modulating the cell inflammatory response during respiratory distress syndrome.


Camel milk inhibits inflammatory angiogenesis via downregulation of proangiogenic and proinflammatory cytokines in mice.
Camel milk has traditionally been used to treat cancer, but this practice awaits scientific scrutiny, in particular its role in tumor angiogenesis, the key step involved in tumor growth and metastasis. We aimed to investigate the effects of camel milk on key components of inflammatory angiogenesis in sponge implant angiogenesis model. Polyester-polyurethane sponges, used as a framework for fibrovascular tissue growth, were implanted in Swiss albino mice and camel milk (25, 50 and 100 mg/kg/day) was administered for 14 days through installed cannula. The implants collected at day 14 post-implantation were processed for the assessment of hemoglobin (Hb), myeloperoxidase (MPO), N-acetylglucosaminidase (NAG), and collagen, which were used as indices for angiogenesis, neutrophil, and macrophage accumulation and extracellular matrix deposition, respectively. Relevant inflammatory, angiogenic, and fibrogenic cytokines were also determined. Camel milk treatment attenuated the main components of the fibrovascular tissue, wet weight, vascularization (Hb content), macrophage recruitment (NAG activity), collagen deposition and the levels of vascular endothelial growth factor (VEGF), interleukin (IL)-1β, IL-6, IL-17, tumor necrosis factor-α, and transforming growth factor-β. A regulatory function of camel milk on multiple parameters of the main components of inflammatory angiogenesis has been revealed, giving insight into the potential therapeutic benefit underlying the anti-cancer actions of camel milk.


Role of lipid peroxidation in dapsone-induced hemolytic anemia.
Dapsone hydroxylamine (DDS-NOH) is a direct-acting hemolytic agent responsible for dapsone-induced hemolytic anemia in the rat. The hemolytic activity of DDS-NOH is associated with the formation of disulfide-linked hemoglobin adducts on membrane skeletal proteins. We have postulated that this membrane protein "damage" is a consequence of DDS-NOH-induced oxidative stress within the red cell and that it serves as the trigger for premature removal of injured but intact red cells from the circulation by splenic macrophages. Oxidative stress has also been associated with the induction of lipid peroxidation, and it is possible that direct damage to the lipoidal membrane may play a role in the premature sequestration of the damaged cells in the spleen. To investigate this possibility, rat and human red cells were incubated with hemolytic concentrations of DDS-NOH and examined for evidence of lipid peroxidation using two independent assays: thiobarbituric acid-reactive substances formation and cis-paranaric acid degradation. Phenylhydrazine, which is known to induce lipid peroxidation in red cells, was used as a positive control. The extent of thiobarbituric acid-reactive substances formation and cis-paranaric acid degradation in DDS-NOH-treated rat and human red cells was not significantly different from that in control cells. In contrast, thiobarbituric acid-reactive substances formation and cis-paranaric acid degradation were significantly increased in red cells treated with hemolytic concentrations of the positive control, phenylhydrazine. These data suggest that lipid peroxidation is not involved in the mechanism underlying dapsone-induced hemolytic anemia.


Field-theoretic simulations in the Gibbs ensemble.
Calculating phase diagrams and measuring the properties of multiple phases in equilibrium is one of the most common applications of field-theoretic simulations. Such a simulation often attempts to simulate two phases in equilibrium with each other in the same simulation box. This is a computationally demanding approach because it is necessary to perform a large enough simulation so that the interface between the two phases does not affect the estimate of the bulk properties of the phases of interest. In this paper, we describe an efficient method for performing field-theoretic simulations in the Gibbs ensemble, a familiar construct in particle-based simulations where two phases in equilibrium with each other are simulated in separate simulation boxes. Chemical and mechanical equilibrium is maintained by allowing the simulation boxes to swap both chemical species and volume. By fixing the total number of each chemical species and the total volume, the Gibbs ensemble allows for the efficient simulation of two bulk phases at equilibrium in the canonical ensemble. After providing the theoretical framework for field-theoretic simulations in the Gibbs ensemble, we demonstrate the method on two two-dimensional model polymer test systems in both the mean-field limit (self-consistent field theory) and in the fluctuating field theory.


Association of cardiometabolic risk factors and hepatic enzymes in a national sample of Iranian children and adolescents: the CASPIAN-III study.
Elevated liver enzymes including alanine aminotransferase (ALT) and aspartate aminotransferase (AST) may be associated with metabolic syndrome (MetS) and cardiovascular disease. We investigated the association of cardiometabolic risk factors and liver enzymes in a nationally representative sample of Iranian children and adolescents. The national study was conducted in the framework of the third survey of Childhood and Adolescence Surveillance and PreventIon of Adult Non-communicable Disease study. Subjects were 3948 students (1942 girls, 67.55% urban, mean age 14.7 ± 2.4 years) who were recruited by multistage random cluster sampling from 27 provincial counties in Iran. Physical examination and laboratory tests were conducted under standard protocols. Participants with elevated serum ALT had higher levels of almost all cardiometabolic risk factors than other participants; this difference was not significant for fasting blood glucose, total cholesterol, and diastolic blood pressure in both sexes, as well as low-density lipoprotein cholesterol in girls. Participants with generalized and abdominal obesity, MetS, elevated blood pressure, triglycerides, and total cholesterol had increased risk for elevated ALT; this risk remained significant after adjusting for sex and age. Low high-density lipoprotein cholesterol was found as a predictor for both elevated ALT (odds ratio 2.182, 95% confidence interval 1.533-3.105) and AST (odds ratio 2.022, 95% confidence interval 1.438-2.844) even after adjusting for all potential confounders. General (B 0.158, SE 0.030) and abdominal obesity (B 0.058, SE 0.029), MetS (B 0.231, SE 0.048), and triglycerides (B 0.094, SE 0.030) were associated with ALT:AST ratio after adjusting for all potential confounders (P < 0.001). We documented strong relations of elevated ALT, AST levels, and ALT:AST ratio with most cardiometabolic risk factors. This relation was independent of anthropometric indexes. Liver enzymes can be considered as a cardiometabolic risk factor from childhood, and as an additional component of the MetS.


[Cytotoxic properties of a recombinant hybrid of the A protein of Staphylococcus aureus with a fragment of exotoxin A of Pseudomonas aeruginosa].
By gene-engineering technique a chimeric protein made up of fragments of Staphylococcus aureus protein A and . Pseudomonas aeruginosa exotoxin A has been constructed. The chimeric protein was shown to preserve features characteristic of its both constituents--it ADP-ribosylates elongation factor 2 and binds to Ig. Cytotoxic properties of the chimeric protein were studied in two model systems. Treatment of target cells in both systems was performed successively with antibodies against corresponding antigens and after washing--with recombinant chimeric toxin which bound to antibodies on the surface of target cells. In the first model system human B-lymphoma cells (Daudi line) carrying Ig molecules on their surface were treated with polyclonal antibodies against human Ig L-chains. In the other system, human T-lymphoma cells (Jurkat line) were treated successively with monoclonal antibodies against cell surface CD5 antigen and further on--with polyclonal antibodies against mouse Ig. In both systems, only a slight inhibition of the target cells' growth was registered. The probable reasons of low cytotoxic activity of the chimeric protein and prospects of increasing it are discussed.


Methylibium aquaticum sp. nov., a betaproteobacterium isolated from a eutrophic freshwater pond.
A freshwater bacterium, designated IMCC1728(T), was isolated from a eutrophic pond. The strain was Gram-negative, chemoheterotrophic and facultatively aerobic, forming non-motile rods that contained poly-beta-hydroxybutyrate granules. Based on 16S rRNA gene sequence comparisons, the most closely related species to strain IMCC1728(T) was Methylibium petroleiphilum (97.0 % similarity). Phylogenetic trees generated using 16S rRNA gene sequences showed that this isolate formed an independent phyletic line of the genus Methylibium clade of the class Betaproteobacteria. The DNA G+C content of the strain was 66.2+/-0.4 mol%. The predominant cellular fatty acid constituents were C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH (43.1 %), C(16 : 0) (20.3 %), C(12 : 0) (13.4 %) and C(10 : 0) 3-OH (7.3 %). The strain contained Q-8 as the predominant ubiquinone. Several phenotypic characteristics, including flagellation, temperature range for growth and carbon source utilization patterns, differentiated strain IMCC1728(T) from species of the genus Methylibium. Therefore, it is proposed that strain IMCC1728(T) represents a novel species, Methylibium aquaticum sp. nov. The type strain is IMCC1728(T) (=KCCM 42364(T)=NBRC 102349(T)).


Evidence for hyperacute rejection of human liver grafts: The case of the canary kidneys.
Sequential liver and kidney transplantation from the same donor was performed in 2 patients. The kidney in Patient 1, which was transplanted after the liver, was hyperacutely rejected and removed 6 hours later. The first liver as well as another liver transplanted 3 days later developed widespread hemorrhagic necrosis. Although the cytotoxic crossmatch of preoperative recipient serum with both donors was negative, patchy widespread IgM and C(1q) deposits were found in all 3 organs. In Patient 2, who had a strongly positive cytotoxic crossmatch with his donor, the liver suffered a massive but reversible injury, while the kidney never functioned. Both patients developed a coagulopathy a few minutes after liver revascularization. The kidneys in these cases had served like the canaries which miners once used to detect a hostile environment and their presence made more understandable how an indolent version of hyperacute rejection of the liver can take place.


Comparison of two infusion rates of antithrombin concentrate in cardiopulmonary bypass surgery.
Antithrombin concentrate (AT) is used to treat heparin resistance (HR) in cardiac surgery. It is usually given slowly due to the fear of anaphylaxis. This may delay cardiac catheterisation and the start of cardiopulmonary bypass (CPB). HR is often defined as the failure to reach or maintain a target activated clotting time (ACT) despite a standard dose of heparin. It is not generally possible to predict which patients will display HR, although there are known risk factors. Routine early administration of AT before heparinisation is probably not cost-effective. Infusing AT relatively quickly after demonstrating HR may be more cost-effective, while not delaying surgery. The aim of this study is to investigate the safety and side effects of a faster infusion of AT. Forty patients undergoing elective heart surgery were included and randomised to two groups in a double-blind fashion. Each group received 1000 IU of AT intravenously (IV). One group received a slow infusion (100 IU/min) before full-dose heparinisation. The other group received a fast infusion (250 IU/min). Haemodynamic and respiratory data were recorded. Any adverse effects were noted. Thrombin-antithrombin, anti-Xa and antithrombin levels in plasma were measured. No anaphylaxis occurred in either group. No differences were found regarding haemodynamics, respiration or laboratory results. Two patients experienced major haemorrhage and recovered; there were two deaths, thought to be unrelated to the study drugs. AT can be infused at a rate of 250 IU/min. This is faster than the current recommendation of 100 IU/min. This rate of infusion allows restricting AT infusion to those patients who display HR, without delaying surgery. Optimal anticoagulant therapy for CPB probably includes point-of-care measurement of ACT and plasma AT and small, but rapid, infusions of AT in heparin-resistant patients.


[Pyrrolidine acetamide as an auxiliary drug in the treatment of cerebral palsy].
Twenty children, with the diagnosis of cerebral palsy (CP) and under classical, physiotherapeutical and pedagogical, treatment, received piracetam (pyrrolidine acetamide) as an auxiliary drug. The goal was to better spasticity, learning and nervous instability problems aiming at better results of over-all treatment of CP. The group that received the drug has been compared to a control group of 20 children treated by the customary treatment only. The comparison showed favourable results for the medicated group. The drug was administered in the dose of 80 mg/kg/day during 10 weeks. The criteria for evaluation have been psychological, clinical, physiotherapeutical and pedagogical. The drug has been given in a new form of presentation: 6% solution for oral use.


Testicular Ectopia in a Child's Anterior Abdominal Wall: A Case Report and Literature Review.
BACKGROUND Ectopic testis is an uncommon congenital anomaly that has been reported in different sites in the body. The anterior abdominal wall is an exceptionally rare variant site for ectopic testis. The purpose of this case report is to highlight the importance of being aware of this rare clinical entity, thereby increasing the chance of preserving an ectopic testis. In addition, this case report shows how a laparoscopic approach is advantageous for better localizing and treating such rare cases, as this is the first reported case of such a condition to be managed with a laparoscopic approach. CASE REPORT This is a rare case of testicular ectopia in an unclassified abdominal wall defect mimicking a Spigelian hernia that occurred in a 1.5-year-old boy. He presented with congenital swelling of the right lower abdominal wall and an empty right hemiscrotum. Diagnostic laparoscopy was performed and the intraoperative findings suggested right testicular ectopia in the anterior abdominal wall defect. The right testis was herniated through an abnormal, unclassified, abdominal wall defect with both a closed inguinal ring and no defect in the semilunar line. Using a laparoscopic approach, the ectopic testis and hernia were managed successfully with primary hernia repair and single-stage orchidopexy. The patient's postoperative recovery was uneventful. At his 10-month follow-up appointment, he had no evidence of hernia recurrence. CONCLUSIONS As demonstrated in this case, laparoscopic surgery, which has not been used in previously reported cases of ectopic testis, aids in proper localization and repair of an anterior abdominal wall defect along with orchidopexy.


Lateral Approach With a Mouth Corner Incision for the Molar Part of Mandibulectomy.
The aim of the present study was to evaluate the functional and esthetic results of marginal mandibulectomy for mandibular cancer using a lateral approach with a simple, straightforward mouth corner incision. Six mandibular cancer patients (2 men, 4 women; age range: 65-80 years; mean age, 73.1 years; all stage I) were treated using this approach. With this approach, the surgical field was widely exposed, and mandibulectomy was performed with sufficient surgical margins. Intraoperative frozen specimens of remaining tissues showed no malignancy in all cases. In all 6 patients, follow-up imaging assessments were obtained, with no local recurrence after 12 to 78 months. An overall functional, physical, and esthetic assessment of oral behavior and oral appearance was made of all patients by the FACT-H&N questionnaire, which showed that functional lip behavior and esthetic lip appearance were not much affected by the present surgical approach, and good quality of life was maintained. Thus, the lateral approach with the mouth corner incision is an effective and useful alternative for the molar part of mandibulectomy.


PGC1α plays a critical role in TWEAK-induced cardiac dysfunction.
Inflammatory cytokines play an important role in the pathogenesis of heart failure. We have recently found the cytokine TWEAK (tumor necrosis factor (TNF)-like weak inducer of apoptosis), a member of the TNF superfamily, to be increased in patients with cardiomyopathy and result in the development of heart failure when overexpressed in mice. The molecular mechanisms underlying TWEAK-induced cardiac pathology, however, remain unknown. Using mouse models of elevated circulating TWEAK levels, established through intravenous injection of adenovirus expressing TWEAK or recombinant TWEAK protein, we find that TWEAK induces a progressive dilated cardiomyopathy with impaired contractile function in mice. Moreover, TWEAK treatment is associated with decreased expression of peroxisome proliferator-activated receptor gamma coactivator-1α (PGC1α) and genes required for mitochondrial oxidative phosphorylation, which precede the onset of cardiac dysfunction. TWEAK-induced downregulation of PGC1α requires expression of its cell surface receptor, fibroblast growth factor-inducible 14 (Fn14). We further find that TWEAK downregulates PGC1α gene expression via the TNF receptor-associated factor 2 (TRAF2) and NFκB signaling pathways. Maintaining PGC1α levels through adenoviral-mediated gene expression is sufficient to protect against TWEAK-induced cardiomyocyte dysfunction. Collectively, our data suggest that TWEAK induces cardiac dysfunction via downregulation of PGC1α, through FN14-TRAF2-NFκB-dependent signaling. Selective targeting of the FN14-TRAF2-NFκB-dependent signaling pathway or augmenting PGC1α levels may serve as novel therapeutic strategies for cardiomyopathy and heart failure.


Interactions of cadmium and aluminum toxicity in their effect on growth and physiological parameters in soybean.
The effect of Al and Cd on the growth, photosynthesis, and accumulation of Al, Cd and plant nutrients in two soybean genotypes were determined using hydroponic culture. There were six treatments: pH 6.5; pH 4.0; pH 6.5+1.0 micromol/L Cd; pH 4.0+1.0 micromol/L Cd; pH 4.0+150 micromol/L Al; pH 4.0+1.0 micromol/L Cd+150 micromol/L Al. The low pH (4.0) and Al treatments caused marked reduction in root length, shoot height, dry weight, chlorophyll content (SPAD value) and photosynthetic rate. Al-sensitive cv. Zhechun 2 accumulated comparatively more Al and Cd in plants than Al-tolerant cv. Liao 1. Compared with pH 6.5, pH 4.0 resulted in significant increase in Cd and Al concentration in plants. Combined application of Cd and Al enhanced their accumulation in roots, but caused a reduction in shoots. The concentrations of all 10 nutrients (P, K, Ca, Mg, Fe, Mn, Cu, Zn and B), except Mo were also increased when plants were exposed to pH lower than pH 6.5. Al addition caused a reduction in the concentration of most nutrients in plant roots and shoots; but K, Mn and Zn in roots were increased. Treatments with Cd alone or together with Al reduced the concentrations of all the plant nutrients in plants. Al-sensitive genotype Zhechun 2 has lower nutrient concentration than Al-tolerant genotype Liao 1. The current findings imply that Al and Cd are synergistic in their effect on plant growth, physiological traits and nutrient uptake.


[Epithelioid sarcoma. Apropos of 3 cases].
The authors present two cases of epithelioid sarcoma of the hand and one of the foot with clinical interest. This recently described tumor (Enzinger, 1970), is relatively rare. It occurs in the hand, forearm, pretibial region and foot and affects principally young adults. It should be emphasized, and this is borne out in the literature, that this tumor may appear perfectly benign and often has a course of long duration. The difficulty of clinical and especially of histologic diagnosis as well as the difficulty of determining the boundaries of extension of the tumor make it necessary to carry out radical surgery (amputation or rarely block excision). Every author agrees that local excision is to be condemned, there being an 85% recurrence rate. Spread of the tumor is by way of the fascial planes and tendon sheaths. Lymphatic and pulmonary metastases occur particularly when there is vascular invasion.


Novel Pathways between Invasiveness Modulators in Breast Cancer Single Cells.
Individual cells are known to behave differently than their whole populations of cells. The present work focused on proteins that control cancer invasiveness. Individual Dicer siRNA knockdown of HER4, CDC42, and E-cadherin decreased MMP1 mRNA levels in SCP2, a cancer single-cell progeny that is highly metastatic to bone and adrenal gland. Individual knockdown of β-catenin, CDC42, HER3, and the γ catalytic subunit of PI3K raised MMP1 mRNA levels in SCP21, a single-cell progeny of the same tumor and patient, with low metastasis to bone and adrenal.


FUS-dependent liquid-liquid phase separation is important for DNA repair initiation.
RNA-binding proteins (RBPs) are emerging as important effectors of the cellular DNA damage response (DDR). The RBP FUS is implicated in RNA metabolism and DNA repair, and it undergoes reversible liquid-liquid phase separation (LLPS) in vitro. Here, we demonstrate that FUS-dependent LLPS is necessary for the initiation of the DDR. Using laser microirradiation in FUS-knockout cells, we show that FUS is required for the recruitment to DNA damage sites of the DDR factors KU80, NBS1, and 53BP1 and of SFPQ, another RBP implicated in the DDR. The relocation of KU80, NBS1, and SFPQ is similarly impaired by LLPS inhibitors, or LLPS-deficient FUS variants. We also show that LLPS is necessary for efficient γH2AX foci formation. Finally, using superresolution structured illumination microscopy, we demonstrate that the absence of FUS impairs the proper arrangement of γH2AX nanofoci into higher-order clusters. These findings demonstrate the early requirement for FUS-dependent LLPS in the activation of the DDR and the proper assembly of DSB repair complexes.


Cocaine inhibits cromakalim-activated K+ currents in follicle-enclosed Xenopus oocytes.
The effect of cocaine on K+ currents activated by the KATP channel opener cromakalim was investigated in follicular cells of Xenopus oocytes. The results indicate that cocaine in the concentration range of 3-500 microM reversibly inhibits cromakalim-induced K+ currents. The IC50 value for cocaine was 96 microM. Inhibition of the cromakalim-activated K+ current by cocaine was noncompetitive and voltage independent. Pretreatment with the Ca2+ chelator BAPTA did not modify the cocaine-induced inhibition of cromakalim-induced K+ currents, suggesting that Ca2+-activated second messenger pathways are not involved in the actions of cocaine. Outward K+ currents activated by the application of 8-Br-cAMP or forskolin were also inhibited by cocaine. The EC50 and slope values for the activation of K+ currents by cromakalim were 184+/-19 microM and 1.14 in the absence of cocaine as compared to 191+/-23 microM and 1.03 in the presence of cocaine (300 microM). Cocaine also blocked K+ currents mediated through C-terminally deleted form of Kir6.2 (KirDeltaC26) in the absence of sulfonylurea receptor with an IC50 value of 87 microM, suggesting that cocaine interacts directly with the channel forming Kir6.2 subunit. Radioligand binding studies indicated that cocaine (100 microM) did not affect the binding characteristics of the KATP ligand, [3H]glibenclamide. These results demonstrate that cromakalim-activated K+ currents in follicular cells of Xenopus oocytes are modulated by cocaine.


Mutations in the genes encoding the transcription factors hepatocyte nuclear factor 1 alpha and 4 alpha in maturity-onset diabetes of the young and hyperinsulinemic hypoglycemia.
Maturity-onset diabetes of the young (MODY) is a monogenic disorder characterized by autosomal dominant inheritance of young-onset (typically <25 years), noninsulin-dependent diabetes due to defective insulin secretion. MODY is both clinically and genetically heterogeneous with mutations in at least 10 genes. Mutations in the HNF1A gene encoding hepatocyte nuclear factor-1 alpha are the most common cause of MODY in most adult populations studied. The number of different pathogenic HNF1A mutations totals 414 in 1,247 families. Mutations in the HNF4A gene encoding hepatocyte nuclear factor-4 alpha are a rarer cause of MODY with 103 different mutations reported in 173 families to date. Sensitivity to treatment with sulfonylurea tablets is a feature of both HNF1A and HNF4A mutations. The HNF4A MODY phenotype has been expanded by the reports of macrosomia in ∼50% of babies, and more rarely, neonatal hyperinsulinemic hypoglycemia. The identification of an HNF1A or HNF4A gene mutation has important implications for clinical management in diabetes and pregnancy, but MODY is significantly underdiagnosed. Current research is focused on identifying biomarkers and developing probability models to identify those patients most likely to have MODY, until next generation sequencing technology enables cost-effective gene analysis for all patients with young onset diabetes.


A Web-Based Intervention (Germ Defence) to Increase Handwashing During a Pandemic: Process Evaluations of a Randomized Controlled Trial and Public Dissemination.
Washing hands helps prevent transmission of seasonal and pandemic respiratory viruses. In a randomized controlled trial (RCT) during the swine flu outbreak, participants with access to a fully automated, digital intervention promoting handwashing reported washing their hands more often and experienced fewer respiratory tract infections than those without access to the intervention. Based on these findings, the intervention was adapted, renamed as "Germ Defence," and a study was designed to assess the preliminary dissemination of the intervention to the general public to help prevent the spread of seasonal colds and flu. This study compares the process evaluations of the RCT and Germ Defence dissemination to examine (1) how web-based research enrollment procedures affected those who used the intervention, (2) intervention usage in the 2 contexts, and (3) whether increased intentions to wash hands are replicated once disseminated. The RCT ran between 2010 and 2012 recruiting participants offline from general practices, with restricted access to the intervention (N=9155). Germ Defence was disseminated as an open access website for use by the general public from 2016 to 2019 (N=624). The process evaluation plan was developed using Medical Research Council guidance and the framework for Analyzing and Measuring Usage and Engagement Data. Both interventions contained a goal-setting section where users self-reported current and intended handwashing behavior across 7 situations. During web-based enrolment, 54.3% (17,511/32,250) of the RCT participants dropped out of the study compared to 36.5% (358/982) of Germ Defence users. Having reached the start of the intervention, 93.8% (8586/9155) of RCT users completed the core section, whereas 65.1% (406/624) of Germ Defence users reached the same point. Users across both studies selected to increase their handwashing in 5 out of 7 situations, including before eating snacks (RCT mean difference 1.040, 95% CI 1.016-1.063; Germ Defence mean difference 0.949, 95% CI 0.766-1.132) and after blowing their nose, sneezing, or coughing (RCT mean difference 0.995, 95% CI 0.972-1.019; Germ Defence mean difference 0.842, 95% CI 0.675-1.008). By comparing the preliminary dissemination of Germ Defence to the RCT, we were able to examine the potential effects of the research procedures on uptake and attrition such as the sizeable dropout during the RCT enrolment procedure that may have led to a more motivated sample. The Germ Defence study highlighted the points of attrition within the intervention. Despite sample bias in the trial context, the intervention replicated increases in intentions to handwash when used "in the wild." This preliminary dissemination study informed the adaptation of the intervention for the COVID-19 health emergency, and it has now been disseminated globally. ISRCTN Registry ISRCTN75058295; https://www.isrctn.com/ISRCTN75058295.


Health Behaviors in Austrian Apprentices and School Students during the COVID-19 Pandemic.
The COVID-19 pandemic has disrupted our daily lives, which in turn has impacted health behaviors. Young people have been particularly affected. This study aimed to assess health behaviors in Austrian apprentices and high school students during the COVID-19 pandemic and whether vaccination willingness is affected by health behaviors. Two online surveys were conducted via REDCap with 1442 apprentices (female: 53.5%, male: 45.4%) from 29 March to 18 May 2021 and 563 school students (female: 79.6%, male: 18.6%) from 19 June to 2 July 2021. The two samples were matched to account for sociodemographic differences and analyses were run on the matched sample. Besides the health behaviors, namely, smoking, alcohol consumption, cannabis consumption, and exercise, health status and vaccination willingness were also assessed. Health behaviors were affected by both education group and gender. Apprentices reported significantly more smoking than high school students and this difference was more pronounced in women (all p < 0.01). Alcohol consumption was higher in apprentices than school students, but only in women (p < 0.01). There was a trend for the two education groups to differ in their cannabis use as well (p = 0.05). Apprentices took part in more weekly exercise (p < 0.0001), but high school students reported better average health status (p < 0.001). When included in the same model, health behaviors did not affect vaccine willingness, but education group did, with high school students showing a higher willingness to receive the COVD-19 vaccine than apprentices. These findings support the argument that education type is an important factor for health behaviors, but this is also mediated by gender. Appropriate interventions for adolescents are needed to prevent adverse health behavior changes following the COVID-19 pandemic.


Cloning and expression in Escherichia coli of a gene, hup, encoding the histone-like protein HU of Bifidobacterium longum.
A genomic DNA library of Bifidobacterium longum ATCC15707 was transfected into an Escherichia coli strain deficient in both HU and IHF, the growth of which is cold-sensitive because of the deficiency in these proteins. Cold-resistant colonies were selected and the DNA was cloned and sequenced. A polypeptide consisted of 93 amino acids, a predicted molecular mass of 9983 Da with an isoelectric point of 10.35, was deduced from an orf in the middle of the DNA fragment. The amino acid sequence was highly similar to HU family proteins, and 26 aas of N terminal was identical to a histone-like protein, HBI, a HU family protein of B. longum. Incapabilities of Mu phage propagation in an E. coli mutant deficient in HU or IHF could be suppressed by DNA bearing this orf. These results showed that the orf is a gene hup encoding HBI, a histone-like protein HU of B. longum.


Quantification of task-dependent cortical activation evoked by robotic continuous wrist joint manipulation in chronic hemiparetic stroke.
Cortical damage after stroke can drastically impair sensory and motor function of the upper limb, affecting the execution of activities of daily living and quality of life. Motor impairment after stroke has been thoroughly studied, however sensory impairment and its relation to movement control has received less attention. Integrity of the somatosensory system is essential for feedback control of human movement, and compromised integrity due to stroke has been linked to sensory impairment. The goal of this study is to assess the integrity of the somatosensory system in individuals with chronic hemiparetic stroke with different levels of sensory impairment, through a combination of robotic joint manipulation and high-density electroencephalogram (EEG). A robotic wrist manipulator applied continuous periodic disturbances to the affected limb, providing somatosensory (proprioceptive and tactile) stimulation while challenging task execution. The integrity of the somatosensory system was evaluated during passive and active tasks, defined as 'relaxed wrist' and 'maintaining 20% maximum wrist flexion', respectively. The evoked cortical responses in the EEG were quantified using the power in the averaged responses and their signal-to-noise ratio. Thirty individuals with chronic hemiparetic stroke and ten unimpaired individuals without stroke participated in this study. Participants with stroke were classified as having severe, mild, or no sensory impairment, based on the Erasmus modification of the Nottingham Sensory Assessment. Under passive conditions, wrist manipulation resulted in contralateral cortical responses in unimpaired and chronic stroke participants with mild and no sensory impairment. In participants with severe sensory impairment the cortical responses were strongly reduced in amplitude, which related to anatomical damage. Under active conditions, participants with mild sensory impairment showed reduced responses compared to the passive condition, whereas unimpaired and chronic stroke participants without sensory impairment did not show this reduction. Robotic continuous joint manipulation allows studying somatosensory cortical evoked responses during the execution of meaningful upper limb control tasks. Using such an approach it is possible to quantitatively assess the integrity of sensory pathways; in the context of movement control this provides additional information required to develop more effective neurorehabilitation therapies.


Long-term lithium carbonate therapy causes hyperparathyroidism.
Twelve patients taking lithium carbonate for two to 13 years were compared with age- and sex-matched controls to determine whether long-term lithium carbonate therapy is associated with alterations in calcium metabolism. As a group and individually, patients had significantly higher levels of serum total calcium, ionized calcium, and parathyroid hormone. When compared with serum calcium levels, patients' parathyroid hormone levels were significantly more likely than those of controls to indicate hyperparathyroidism. Lithium carbonate-induced mild hyperparathyroidism appears to be more common than had previously been suspected.


The impact of surgical approach and urinary diversion on patient outcomes in pediatric pyeloplasty.
Recent comparisons of the impact of the surgical approach on pediatric pyeloplasty outcomes have generally incorporated a form of internal ureteral drainage. We hypothesized that the surgical approach does not affect outcomes in children who undergo unstented pyeloplasty and stenting offers no long-term benefit in those with pediatric pyeloplasty. After receiving institutional review board approval we examined the records of all children who underwent initial pyeloplasty from December 2001 to December 2009. We compared unstented and stented pyeloplasties, and each surgical approach in the unstented group. During the study period 367 pyeloplasties were performed, including 231 unstented and 136 stented procedures. When comparing unstented to stented pyeloplasties, there was no difference in the complication or failure rate. Of unstented pyeloplasties 71, 67 and 93 were done using a transperitoneal laparoscopic approach, a flank approach and dorsal lumbotomy, respectively. There were 5 failures, of which 2 were laparoscopic, 2 used a flank approach and 1 used dorsal lumbotomy (p = 0.51). A total of 31 patients, including 10 treated with a laparoscopic approach, 3 with a flank approach and 18 with dorsal lumbotomy (p = 0.02), required second procedures, of which 24 were temporary drainage for a urine leak. Univariate analysis of factors associated with secondary procedures in the unstented pyeloplasty group showed that only surgical approach was significant (p = 0.05). In pediatric pyeloplasty there is no significant difference in outcome between stented and unstented repairs. In unstented repairs complications may vary by surgical approach. Regardless of the approach unstented pyeloplasty is safe and effective in the pediatric population.


Association of the US Affordable Care Act With Out-of-Pocket Spending and Catastrophic Health Expenditures Among Adult Patients With Traumatic Injury.
Trauma is an expensive and unpredictable source of out-of-pocket spending for American families. The Patient Protection and Affordable Care Act (ACA) sought to improve financial protection by expanding health insurance coverage, but its association with health care spending for patients with traumatic injury remains largely unknown. To evaluate the association of ACA implementation with out-of-pocket spending, premiums, and catastrophic health expenditures (CHE) among adult patients with traumatic injury. Data from a nationally representative sample of US adults aged 19 to 64 years who had a hospital stay or emergency department visit for a traumatic injury from January 2010 to December 2017 were analyzed using the Medical Expenditure Panel Survey. Multivariable generalized linear models were used to evaluate changes in spending after ACA implementation. Additionally, 4 income subgroups were evaluated based on ACA thresholds for program eligibility: lowest-income patients (earning 138% or less of the federal poverty level [FPL]), low-income patients (earning 139% to 250% of the FPL), middle-income patients (earning 251% to 400% of the FPL), and high-income patients (earning more than 400% of the FPL). Data were analyzed from February to December 2019. Implementation of the ACA, beginning January 1, 2014. Out-of-pocket spending, premium spending, out-of-pocket plus premium spending, and likelihood of experiencing CHE, defined as out-of-pocket plus premium spending exceeding 19.5% of family income. Of the 6288 included patients, 2995 (weighted percentage, 51.3%) were male, and the mean (SD) age was 41.4 (12.8) years. Implementation of the ACA was associated with 31% lower odds of CHE (adjusted odds ratio, 0.69; 95% CI, 0.54 to 0.87; P = .002). Changes were greatest in lowest-income patients, who experienced 30% lower out-of-pocket spending (adjusted percentage change, -30.4%; 95% CI, -46.6% to -9.4%; P = .01), 26% lower out-of-pocket plus premium spending (adjusted percentage change, -26.3%; 95% CI, -41.0% to -8.1%; P = .01), and 39% lower odds of CHE (adjusted odds ratio, 0.61; 95% CI, 0.44 to 0.84; P = .002). Low-income patients experienced decreased out-of-pocket spending and out-of-pocket plus premium spending but no changes in CHE, while middle-income and high-income patients experienced no significant changes in any spending outcome. In the post-ACA period, 1 in 11 of all patients with traumatic injury and 1 in 5 with the lowest incomes continued to experience CHE each year. Implementation of the ACA was associated with improved financial protection for US adults with traumatic injury, especially lowest-income individuals targeted by the law's Medicaid expansions. Despite these gains, injured patients remain at risk of financial strain.


Clinical significance of MCP-1 levels in BALF and serum in patients with interstitial lung diseases.
It has previously been reported that the expression of monocyte chemoattractant protein-1 (MCP-1) in the lung tissues of patients with idiopathic pulmonary fibrosis (IPF) was different from that in the tissues of patients with other interstitial lung diseases (ILDs). The aim of this study was to determine whether this difference reflects the amount of MCP-1 in the bronchoalveolar lavage fluid (BALF) or serum of patients with ILD, and whether such a correlation, if it exists, is clinically useful. MCP-1 concentrations in the BALF and sera were evaluated in 86 patients with ILDs including IPF, acute interstitial pneumonia, interstitial pneumonia with collagen vascular disease (IP-CVD), chronic interstitial pneumonia (CIP), bronchiolitis obliterans-organizing pneumonia, sarcoidosis, hypersensitivity pneumonitis, and in 10 normal healthy volunteers who were controls (NC). BALF MCP-1 levels were significantly elevated in the IPF, IP-CVD, CIP and sarcoidosis groups compared with the NC group. The level in the IPF group was significantly higher than that in any other patient group. Serum MCP-1 levels in the IPF, IP-CVD, CIP and sarcoidosis groups were significantly higher than the NC group. No statistical difference was found in serum MCP-1 levels between the IPF, IP-CVD and CIP groups. BALF MCP-1 levels were significantly higher than serum MCP-1 levels in the IPF group and lower than in the IP-CVD and CIP groups. Serum MCP-1 levels correlated with the clinical course of ILD treated with corticosteroid therapy. These results show that measurement of monocyte chemoattractant protein-1 levels in both bronchoalveolar lavage fluid and serum may be helpful in discriminating idiopathic pulmonary fibrosis from other types of interstitial lung disease and that monitoring of serum monocyte chemoattractant protein-1 may be useful for predicting the clinical course of interstitial lung diseases.


High-energy electrons produced in subpicosecond laser-plasma interactions from subrelativistic laser intensities to relativistic intensities.
The characteristics of the forward hot electrons produced by subpicosecond laser-plasma interactions are studied for different laser polarizations at laser intensities from subrelativistic to relativistic. The peak of the hot electron beam produced by p-polarized laser beam shifts to the laser propagation direction from the target normal direction as the laser intensity reaches the relativistic. For s-polarized laser pulse, hot electrons are mainly directed to the laser axis direction. The temperature and the maximum energy of hot electrons are much higher than that expected by the empirical scaling law. The energy spectra of the hot electrons evolve to be a single-temperature structure at relativistic laser intensities from the two-temperature structure at subrelativistic intensities. For relativistic laser intensities, the forward hot electrons are less dependent on the laser polarization under the laser conditions. The existing of a preplasma formed by the laser amplified spontaneous emission pedestal plays an important role in the interaction. One-dimensional particle-in-cell simulations reproduce the most characteristics observed in the experiment.


Field Theory of Charge Sharpening in Symmetric Monitored Quantum Circuits.
Monitored quantum circuits (MRCs) exhibit a measurement-induced phase transition between area-law and volume-law entanglement scaling. MRCs with a conserved charge additionally exhibit two distinct volume-law entangled phases that cannot be characterized by equilibrium notions of symmetry-breaking or topological order, but rather by the nonequilibrium dynamics and steady-state distribution of charge fluctuations. These include a charge-fuzzy phase in which charge information is rapidly scrambled leading to slowly decaying spatial fluctuations of charge in the steady state, and a charge-sharp phase in which measurements collapse quantum fluctuations of charge without destroying the volume-law entanglement of neutral degrees of freedom. By taking a continuous-time, weak-measurement limit, we construct a controlled replica field theory description of these phases and their intervening charge-sharpening transition in one spatial dimension. We find that the charge fuzzy phase is a critical phase with continuously evolving critical exponents that terminates in a modified Kosterlitz-Thouless transition to the short-range correlated charge-sharp phase. We numerically corroborate these scaling predictions also hold for discrete-time projective-measurement circuit models using large-scale matrix-product state simulations, and discuss generalizations to higher dimensions.


Nonlinear Kramers equation associated with nonextensive statistical mechanics.
Stationary and time-dependent solutions of a nonlinear Kramers equation, as well as its associated nonlinear Fokker-Planck equations, are investigated within the context of Tsallis nonextensive statistical mechanics. Since no general analytical time-dependent solutions are found for such a nonlinear Kramers equation, an ansatz is considered and the corresponding asymptotic behavior is studied and compared with those known for the standard linear Kramers equation. The H-theorem is analyzed for this equation and its connection with Tsallis entropy is investigated. An application is discussed, namely the motion of Hydra cells in two-dimensional cellular aggregates, for which previous measurements have verified q-Gaussian distributions for velocity components and superdiffusion. The present analysis is in quantitative agreement with these experimental results.


Alveolar ridge preservation using resorbable bioactive ceramic composite: a histological study.
The purpose of this study was to histologically evaluate newly generated vital bone using porous granules of bioactive and resorbable silica-calcium phosphate nanocomposite (SCPC) in extraction sockets. Six patients with a non-restorable maxillary central incisor requiring extraction followed by implant placement participated in the study. Extraction sockets were grafted with granules of SCPC. After 6 months, a bone core sample was retrieved from the center of the healed socket for histologic analysis, and dental implants were placed. Alveolar bone width was clinically assessed immediately after tooth extraction and 6 months after bone grafting, at the time of implant placement. Alveolar bone height was radiographically assessed immediately after tooth extraction and 6 months after extraction. Histomorphometric analyses of sockets grafted with SCPC for 6 months revealed 46.8% +/- 14% new vital bone and 2.5% +/- 1.5% graft material remnants. In these sockets, the mean bone height resorption over the 6-month period of healing was 1.6 mm +/- 1.5 mm. The mean bone width resorption of 2 mm +/- 0.7 mm was found at the bone crest. The results of this study suggest that SCPC graft material reduces the amount of change in alveolar ridge dimensions after tooth extraction and facilitates the regeneration of new vital bone.


Long-term DHEA substitution in female adrenocortical failure, body composition, muscle function, and bone metabolism: a randomized trial.
Adrenal derived androgens are low in women with adrenal failure. The physiological consequences of substitution therapy are uncertain. To investigate the effects of DHEA substitution in women with adrenal failure on body composition, fuel metabolism, and inflammatory markers. DESIGN, PARTICIPANTS AND INTERVENTION: In this study, ten female patients (median age 38.5 years, range 28-52) with adrenal failure were treated with DHEA 50 mg for 6 months in a double-blind, randomized, placebo-controlled, and crossover study. The participants underwent dual-energy X-ray absorptiometry (DXA) scan, computed tomography scan of abdominal fat, indirect calorimetry, bicycle ergometry, muscle and fat biopsies, and blood samples. Baseline androgens were normalized to fertile range during active treatment. Anthropometric data were unaffected, but lean body mass (LBM) slightly increased compared with placebo (delta LBM (kg) placebo versus DHEA: -0.48±6.1 vs 1.6±3.4, P=0.02) with no alterations in total or abdominal fat mass. PTH increased with DHEA, but no significant changes were observed in other bone markers or in bone mineral content. The mRNA levels of markers of tissue inflammation (adiponectin, interleukin 6 (IL6), IL10, monocyte chemoattractant protein 1, and tumor necrosis factor α) in fat and muscle tissue were unaffected by DHEA treatment, as was indirect calorimetry and maximal oxygen uptake. A high proportion of self-reported seborrheic side effects were recorded (60%). In female adrenal failure, normalization of androgens with DHEA 50 mg for 6 months had no effects on muscle, fat, and bone tissue and on fuel metabolism in this small study. A small increase in LBM was observed. Treatment was associated with a high frequency of side effects.


Association between Motilin Receptor Gene Haplotypes and Growth Traits in Japanese Hinai-dori Crossbred Chickens.
We previously identified quantitative trait loci (QTL) for body weight and average daily gain in a common region between ADL0198 (chr 1: 171.7 Mb) and ABR0287 (chr 1: 173.4 Mb) on chicken chromosome 1 in an F2 resource population produced by crossing low- and high-growth lines of the Hinai-dori breed. Motilin receptor (MLNR) is a candidate gene affecting growth traits in the region. In this study, we genotyped polymorphisms of the MLNR gene and investigated its association with growth traits in a Hinai-dori F2 intercross population. All the exons of the MLNR gene in the parental population were subjected to PCR amplification, nucleotide sequenced and haplotypes identified. To distinguish resultant diplotype individuals in the F2 population, a mismatch amplification mutation assay was performed. Three haplotypes (Haplotypes 1-3) were accordingly identified. Six genotypes produced by the combination of three haplotypes (Haplotype 1, 2, and 3) were examined in order to identify associations between MLNR haplotypes and growth traits. The data showed that Haplotype 1 was superior to Haplotype 2 and 3 in body weight at 10 and 14 weeks of age, average daily gain between 4 and 10 weeks, 10 and 14 weeks, and 0 and 14 weeks of age in female in F2 females. It was concluded that MLNR is a useful marker of growth traits and could be used to develop strategies for improving growth traits in the Hinai-dori breed.


The production of communication signals at the air-water and water-substrate boundaries.
The 2 interfaces of the aquatic environment, the boundary between air-water and water-substrate, have distinctive physical characteristics that facilitate the production of communication signals. Recent evidence suggests that animals living on or near these boundaries use the interface to generate signals in 2 ways: (a) by producing a signal that propagates along the interface or (b) by producing a signal at the interface that is transmitted and detected within 1 of the component media. By examining the diversity of behaviors used to produce signals at these boundaries, the authors illustrate how human perception of these environments may cause researchers to incorrectly assume the environmental context of signal-generating behaviors and overlook modalities of communication pertinent to the animal.


Auxin regulates distal stem cell differentiation in Arabidopsis roots.
The stem cell niche in the root meristem is critical for the development of the plant root system. The plant hormone auxin acts as a versatile trigger in many developmental processes, including the regulation of root growth, but its role in the control of the stem cell activity remains largely unclear. Here we show that local auxin levels, determined by biosynthesis and intercellular transport, mediate maintenance or differentiation of distal stem cells in the Arabidopsis thaliana roots. Genetic analysis shows that auxin acts upstream of the major regulators of the stem cell activity, the homeodomain transcription factor WOX5, and the AP-2 transcription factor PLETHORA. Auxin signaling for differentiation of distal stem cells requires the transcriptional repressor IAA17/AXR3 as well as the ARF10 and ARF16 auxin response factors. ARF10 and ARF16 activities repress the WOX5 transcription and restrict it to the quiescent center, where WOX5, in turn, is needed for the activity of PLETHORA. Our investigations reveal that long-distance auxin signals act upstream of the short-range network of transcriptional factors to mediate the differentiation of distal stem cells in roots.


Circ_0004904 suppresses trophoblast cell proliferation, invasion and migration in preeclampsia via upregulating the expression of DNA damage inducible alpha by interacting with miR-19a-3p.
Preeclampsia (PE) is the most common complication in the pregnancy of women. PE progression was found to be associated with dysregulated circular RNAs (circRNAs), and we aimed to explore the pathological mechanism with circ_0004904 in PE. The circ_0004904, microRNA-19a-3p (miR-19a-3p) and DNA damage inducible alpha (GADD45A) were quantified via reverse transcription-quantitative polymerase chain reaction assay. Trophoblast cell behaviors were examined by cell viability using Cell Counting Kit-8 assay, cell proliferation using EdU assay, cell apoptosis using flow cytometry, cell invasion using transwell assay and migration using wound healing assay. Western blot was used for protein analysis of epithelial mesenchymal transition (EMT) and GADD45A. Dual-luciferase reporter assay and RNA immunoprecipitation assay were used for target interaction analysis. The circ_0004904 upregulation was detected in placenta tissues from PE patients. Trophoblast cell proliferation, invasion, migration and EMT were repressed but cell apoptosis was promoted after overexpression of circ_0004904. Circ_0004904 acted as a miR-19a-3p sponge in trophoblast cells, and all regulatory effects of circ_0004904 on trophoblast cell behaviors were reversed by miR-19a-3p upregulation. The miR-19a-3p directly targeted GADD45A and miR-19a-3p downregulation inhibited trophoblast cell development through elevating the GADD45A level. Moreover, circ_0004904 enhanced the expression of GADD45A via sponging miR-19a-3p. Our results elucidated that circ_0004904 reduced proliferation and cell motility of trophoblast cells via the miR-19a-3p-mediated GADD45A level elevation, indicating the involvement of circ_0004904/miR-19a-3p/GADD45A in PE progression.


Comparative proteomics of human embryonic stem cells and embryonal carcinoma cells.
Pluripotent human embryonic stem cells (ESCs) can be differentiated in vitro into a variety of cells which hold promise for transplantation therapy. Human embryonal carcinoma cells (ECCs), stem cells of human teratocarcinomas, are considered a close but malignant counterpart to human ESCs. In this study, a comprehensive quantitative proteomic analysis of ESCs and ECCs was carried out using the iTRAQ method. Using two-dimensional LC and MS/MS analyses, we identified and quantitated approximately 1800 proteins. Among these are proteins associated with pluripotency and development as well as tight junction signaling and TGFbeta receptor pathway. Nearly approximately 200 proteins exhibit more than twofold difference in abundance between ESCs and ECCs. Examples of early developmental markers high in ESCs include beta-galactoside-binding lectin, undifferentiated embryonic cell transcription factor-1, DNA cytosine methyltransferase 3beta isoform-B, melanoma antigen family-A4, and interferon-induced transmembrane protein-1. In contrast, CD99-antigen (CD99), growth differentiation factor-3, cellular retinoic acid binding protein-2, and developmental pluripotency associated-4 were among the highly expressed proteins in ECCs. Several proteins that were highly expressed in ECCs such as heat shock 27 kDa protein-1, mitogen-activated protein kinase kinase-1, nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor like-2, and S100 calcium-binding protein-A4 have also been attributed to malignancy in other systems. Importantly, immunocytochemistry was used to validate the proteomic analyses for a subset of the proteins. In summary, this is the first large-scale quantitative proteomic study of human ESCs and ECCs, which provides critical information about the regulators of these two closely related, but developmentally distinct, stem cells.


Intussusception in Johannesburg. A review of 81 cases.
The difference in presentation between two different social groups of children with intussusception was investigated. The sex ratio of the two groups was different. The underprivileged children presented later in the disease and consequently required bowel resection more frequently. The introduction of intravenously administered ketamine increased the barium reduction rate significantly.


An oral sensitization model in Brown Norway rats to screen for potential allergenicity of food proteins.
We developed an oral sensitization protocol for food proteins for the rat. Young Brown Norway (BN) rats were exposed to 1 mg ovalbumin (OVA) by daily gavage dosing for 42 days without the use of an adjuvant. OVA-specific IgE and IgG responses were determined by ELISA. On an oral challenge with OVA some clinical symptoms of food allergy-like effects on the respiratory system, blood pressure, and permeability of the gastrointestinal barrier were studied. In addition, BN rats were orally exposed to a total hen egg white protein (HEW) extract and cow's milk (CM) and the specificities of induced antibody responses were compared with the specificities of antibodies in sera from egg- and milk-allergic patients using immunoblotting. Animals orally exposed to the allergens developed specific IgE and IgG antibodies which recognized the same proteins compared with antibodies from egg- or CM-allergic patients. Among the various clinical symptoms of food allergy, gut permeability was increased after an oral challenge. In addition, some animals demonstrated a temporary decrease in breathing frequency or systolic blood pressure. The results obtained show that the Brown Norway rat is a suitable animal model for inducing specific IgG and IgE responses on daily intragastric dosing of OVA without the use of an adjuvant. Moreover, local immune-mediated effects on oral challenge are observed. The observation that enterally exposed BN rats and food-allergic patients demonstrate antibody responses to a comparable selection of proteins on exposure to different protein mixtures (HEW and CM) further supports the suitability of the BN rat as an animal model for food allergy research and for the study of the allergenicity of (novel) food proteins.


Secreted Bacterial Effectors and Host-Produced Eiger/TNF Drive Death in aSalmonella-Infected Fruit Fly.
Death by infection is often as much due to the host's reaction as it is to the direct result of microbial action. Here we identify genes in both the host and microbe that are involved in the pathogenesis of infection and disease in Drosophila melanogaster challenged with Salmonella enterica serovartyphimurium (S. typhimurium). We demonstrate that wild-typeS. typhimurium causes a lethal systemic infection when injected into the hemocoel of D. melanogaster. Deletion of the gene encoding the secreted bacterial effect or Salmonella leucine-rich (PslrP)changes an acute and lethal infection to one that is persistent and less deadly. We propose a model in which Salmonella secreted effectors stimulate the fly and thus cause an immune response that is damaging both to the bacteria and, subsequently, to the host. In support of this model, we show that mutations in the fly gene eiger, a TNF homolog, delay the lethality of Salmonella infection. These results suggest that S. typhimurium-infected flies die from a condition that resembles TNF-induced metabolic collapse in vertebrates. This idea provides us with a new model to study shock-like biology in a genetically manipulable host. In addition, it allows us to study the difference in pathways followed by a microbe when producing an acute or persistent infection.


EuroQol-5 dimensions utility gain according to British and Swedish preference sets in rheumatoid arthritis treated with abatacept, rituximab, tocilizumab, or tumour necrosis factor inhibitors: a prospective cohort study from southern Sweden.
The development of EuroQol-5 dimensions (EQ-5D) utility over time in rheumatoid arthritis (RA) patients, treated with biologics other than tumour necrosis factor inhibitors (TNFi), based on the standard British (UK) and the new Swedish (SE) EQ-5D preference sets, has not been previously described. Demographics, core set data, EQ-5D utility, and treatment characteristics for patients with established RA, receiving biologics in southern Sweden from January 2006 to March 2014, were retrieved from observational databases. Theoretical, UK, and experience-based, SE, EQ-5D mean utilities were plotted over time. Data regarding 2418 treatment courses with abatacept (ABA, n = 100), rituximab (RTX, n = 230), tocilizumab (TOC, n = 121), or TNFi (n = 1967) were included in the analysis. Patients starting TNFi treatment, as expected, had shorter disease duration and less previous biologics. Baseline utilities of patients commencing ABA and TOC, but not RTX, were also lower than in the TNFi group. Following treatment initiation, rapid utility improvements were seen with all therapies, reaching plateaus after approximately 1.5 months, and then remaining fairly stable throughout follow-up in patients adhering to therapy. SE utilities were consistently higher than UK, with baseline values at around 0.7 leaving little room for improvement. ABA, RTX, TOC, and TNFi treatments were all associated with favourable EQ-5D utility developments in RA patients adhering to therapy. The compression of the experience-based SE preference set towards higher utilities may compromise its ability to detect between-group differences in quality-adjusted life-years, thus making cost-effectiveness harder to demonstrate in cost-utility analyses applying this preference set, rather than the standard UK.


COPD patients perspectives at the time of diagnosis: a qualitative study.
To gain an understanding of patients perspectives and perceptions of chronic obstructive pulmonary disease (COPD) at the time of diagnosis. A qualitative study using grounded theory. Ten patients in primary care in Sweden, newly diagnosed with COPD or with suspected COPD, were interviewed. The analysis created a process model with a core category "Consequences of smoking" and main categories "Shame", "Appearance of symptoms", "Adaptation", "Reflection", and "Action". "Restrictions in physical capacity" was a key indicator of evolving disease and "Getting a diagnosis" was crucial for the patient. The COPD patient needs a clear diagnosis at an early stage. It is important to seize the opportunity when the presumptive COPD patient is receptive towards support and further action. To detect and support the patient, health professionals must be aware of minor symptoms and underlying mechanisms of possible shame.


[Effects of collagen type ⅩⅦ α1 on epidermal stem cells in aging skin and the microRNA intervention mechanism].
Objective: To investigate the expression and function of collagen type ⅩⅦ α1 (COL17α1) in aging mouse skin and its effect on the stemness and proliferation of human epidermal stem cells (ESCs), and to explore the mechanism of related microRNA (miR) in intervening the expression of COL17α1 of human ESC. Methods: The method of experimental research was used. Twelve 2-month-old (young) and twelve 24-month-old (aged) male C57BL/6J mice were selected, and full-thickness skin samples from their upper back were taken for follow-up detection. After hematoxylin-eosin staining of the full-thickness skin samples of young mice and aged mice, the structure of the epidermis was observed and the thickness of the epidermis was measured; the morphology of epidermal basement membrane and hemidesmosomes were observed by transmission electron microscopy, and the hemidesmosomes were counted; the mRNA and protein expressions of COL17α1 were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting respectively, and the protein expression and distribution of COL17α1 was observed and detected by immunofluorescence method. The fresh foreskin tissue discarded after surgery was obtained from 3 healthy men aged 20-30 years who underwent circumcision at the Fourth Medical Center of PLA General Hospital, ESCs were extracted and well-grown cells were wsed for follow-up experiments. According to the random number table (the same grouping method below), ESCs were divided into blank control group, transfection reagent control group, empty vector plasmid group, and COL17α1 knockdown plasmid group with corresponding treatment. After 48 hours of culture, the mRNA expression of COL17α1 was detected by real-time fluorescent quantitative RT-PCR, the protein expressions of COL17α1 and cytokeratin 14 (CK14) were detected by Western blotting, and the cell proliferation level was detected by cell counting kit 8. miRs that might act on the 3' non-coding region of COL17α1 mRNA were screened through DIANA, miRTarBase, miRNAMap, TargetScan, and microRNA databases. The ESCs were divided into negative control group transfected with miR mimic negative control and each miR mimic group transfected with each of the previously screened miR mimics. Forty-eight hours after transfection, the protein expression of COL17α1 was detected by Western blotting. Based on the sequencing data set GSE114006 in Gene Expression Omnibus (GEO), the GEO2R tool was used to statistically analyze the expression of the previously screened miRs that could cause the reduction of COL17α1 protein expression in the skin of 30 young (18-25 years old) and 30 elderly (>70 years old) human skins. The full-thickness skin samples of young mice and aged mice were taken, and the expressions of increased miRs in the aforementioned aged human skin were detected by real-time fluorescent quantitative RT-PCR. Two batches of human ESCs were taken, the first batch was divided into COL17α1 wild type+miR-203b-3p negative control group and COL17α1 wild type+miR-203b-3p mimic group, and the second batch was divided into COL17α1 mutant+miR-203b-3p negative control group and COL17α1 mutant+miR-203b-3p mimic group. Each group of ESC was transfected with corresponding sequences respectively. Forty-eight hours later, the luciferase reporter gene detection kit was used to detect the gene expression level of COL17α1. The number of samples in the tissue experiment was 6, and the number of samples in the cell experiment was 3. Data were statistically analyzed with independent sample t test, one-way analysis of variance, least significant difference test or Dunnett's test, Mann-Whitney U test or Kruskal-Wallis H test. Results: Compared with those of young mice, the boundary between the epidermis and the dermis of the aged mice skin was blurred and the cell layers were less, and the thickness of epidermis was significantly thinner (Z=-2.88, P<0.01); the morphology of basement membrane was discontinuous, with less unevenly distributed hemidesmosomes at the epidermis-dermis junction, and the number of hemidesmosomes was significantly reduced (Z=-2.91, P<0.01); the mRNA and protein expression levels of COL17α1 in the skin of aged mice were significantly decreased (with t values of 10.61 and 6.85, respectively, P<0.01). Compared with those of young mice, the protein expression of COL17α1 in the basal layer of epidermis and the bulb of hair follicle in the skin of aged mice was significantly decreased (Z=-2.24, P<0.05). After 48 hours of culture, the protein expression levels of COL17α1 in ESCs of blank control group, transfection reagent control group, empty vector plasmid group, and COL17α1 knockdown plasmid group were 1.00±0.27, 1.12±0.21, 1.13±0.23, and 0.42±0.18, respectively. Compared with those of blank control group, the mRNA and protein expression levels of COL17α1, the protein expression level of CK14, and the proliferation level of ESCs in transfection reagent control group and empty vector plasmid group did not change significantly (P>0.05), while these indexes in COL17α1 knockdown plasmid group were significantly decreased (P<0.05 or P<0.01). miR-203a-3p, miR-203b-3p, miR-512-5p, miR-124-3p, miR-28-5p, miR-590-3p, and miR-329-5p might bind to the 3' non-coding region of COL17α1 mRNA. Forty-eight hours after transfection, compared with 1.000±0.224 in negative control group, the protein expression level of COL17α1 in ESCs of miR-329-5p mimic group, miR-203b-3p mimic group, and miR-203a-3p mimic group decreased significantly (0.516±0.188, 0.170±0.025, and 0.235±0.025, with t values of 3.17, 5.43, and 5.07, respectively, P<0.05 or P<0.01). Only the expression level of miR-203b-3p in the skin of the elderly was significantly higher than that of the young (t=3.27, P<0.01). The expression level of miR-203b-3p in the skin of aged mice was significantly higher than that of young mice (Z=-2.88, P<0.01). Forty-eight hours after transfection, the gene expression level of COL17α1 in ESCs of COL17α1 wild type+miR-203b-3p mimic group was significantly lower than that of COL17α1 wild type+miR-203b-3p negative control group (t=7.66, P<0.01). The gene expression level of COL17α1 in ESCs of COL17α1 mutant+miR-203b-3p mimic group was similar to that of COL17α1 mutant+miR-203b-3p negative control group (P>0.05). Conclusions: The mRNA and protein expression levels of COL17α1 decrease with age increasing in mice, which may lead to the detachment of mouse ESC from the epidermal basement membrane. Decreased expression of COL17α1 can inhibit the expression of CK14 and ESC proliferation, which may be responsible for the thinning of the epidermis and slower wound healing in aged human skin. The increased expression of miR-203b-3p in aged mouse skin can target and bind to the 3' non-coding region of COL17α1 mRNA, hindering the post-transcriptional translation process, thus resulting in decreased COL17α1 protein expression.


Sodium-dependent suppression of gamma-aminobutyric-acid-gated chloride currents in internally perfused frog sensory neurones.
1. The effects of the Na+ electrochemical potential gradient on gamma-aminobutyric acid (GABA)-induced Cl- currents (ICl) in frog sensory neurones were studied, using a suction pipette technique with which internal perfusion can be accomplished under current- and voltage-clamp conditions. 2. Under current clamp, the depolarizing response to GABA decreased in the presence of external Na+. A similar external Na+-dependent reduction in the GABA-induced inward ICl was observed under voltage clamp. The reversal potential of GABA-induced ICl (EGABA) was nearly equal to the Cl- equilibrium potential (ECl), irrespective of the presence or absence of external Na+. 3. Varying the Na+ influx by changing the holding membrane potential (VH) altered the GABA response: the GABA-induced ICl decreased progressively as VH became more negative. 4. The effects of changing the external and internal Na+ concentrations ([Na+]o and [Na+]i) on the GABA-induced ICl were also studied. Increasing [Na+]o at a constant [Na+]i reduced this current while increasing [Na+]i at a fixed [Na+]o facilitated it. 5. A high temperature coefficient of about 3 was estimated with respect to the percentage reduction in GABA-induced ICl due to [Na+]o. 6. These results indicate that the [Na+]o-dependent suppression of GABA-induced ICl was mediated chiefly by the uptake of GABA subserved by a Na-GABA co-transport mechanism. 7. GABA dose-response measurements were made with and without external Na+. The [Na+]o-induced suppression was more pronounced in relative amount at lower concentrations and in absolute amount at intermediate concentrations. Analysis of these data indicates, however, that the Na+-coupled GABA influx kept increasing at GABA concentrations high enough to nearly saturate GABA-induced ICl, and the same saturating level was observed as in the Na+-free case. This indicates that the electrogenic co-transport current was much smaller so that our measurements of GABA-induced ICl' were contaminated very little. Thus, the present method based on recording of GABA-induced ICl was legitimate for the analysis of the Na-GABA co-transport. 8. By analysing the [Na+]o-dependent suppression of GABA-induced ICl, the stoichiometric ratio of the underlying co-transport was estimated to be one: one Na+ ion per GABA molecule. 9. The ICl induced by GABA agonists such as beta-alanine, taurine, l-GABOB (l-gamma-amino-beta-hydroxybutyric acid) and muscimol was not affected by the amount of external Na+ present, suggesting difference in the affinity between receptor and transport carrier.


Financial performance of the teaching pharmacies in Isfahan: an economic evaluation.
Teaching pharmacies are amongst the important cornerstones of a healthcare system for drug supplying, pharmacy education and pharmacy practice research. Assessment of the Iranian healthcare system costs shows that after personnel charges, drug outlay is the second expensive factor. This great financial mass requires integral audit and management in order to provide costumers satisfaction in addition to financial viability. Teaching pharmacies are required to realize financial viability as well as providing several educational and drug servicing goals, which makes microeconomic analysis important. The aim of this study was to evaluate the financial performance of the teaching pharmacies affiliated with the Isfahan University of Medical Sciences (with the abrreviated names as: SHM, ISJ, AZH for the confidentialiy of the financial data). This is a descriptive and cross-sectional study done in 2008. The target pharmacies of this study were all the 3 teaching pharmacies affiliated with the Isfahan University of Medical Sciences. The data collecting template was prepared using the standard scientific methods according to the goals of this research The goals also nominated necessary items needed in economic profit evaluation. The data collection template was completed by reference to the teaching pharmacies financial documents and reports, used as a base for calculating the total income and the total costs in 2007-2008 financial year. The difference between these two balances showed the value of profits or loss. The profit/cost ratio was also calculated, using the proportion of the total income to the total costs. The collected data was statistically analyzed using the Excel software (Microsoft 2007). For the financial year 2007-2008, the difference between the total income and the total costs was -831.6 million Rials (excess costs to income) for the SHM pharmacy, + 25.4 billion Rials for the ISJ pharmacy and -429.5 million Rials for the AZH pharmacy. According to our findings there is a strong requirement to improve the financial performance of all the three teaching pharmacies while maintaining a high standardard of teaching and educational affairs.


Early warning of cotton bollworm resistance associated with intensive planting of Bt cotton in China.
Transgenic crops producing Bacillus thuringiensis (Bt) toxins kill some key insect pests, but evolution of resistance by pests can reduce their efficacy. The predominant strategy for delaying pest resistance to Bt crops requires refuges of non-Bt host plants to promote survival of susceptible pests. To delay pest resistance to transgenic cotton producing Bt toxin Cry1Ac, farmers in the United States and Australia planted refuges of non-Bt cotton, while farmers in China have relied on "natural" refuges of non-Bt host plants other than cotton. Here we report data from a 2010 survey showing field-evolved resistance to Cry1Ac of the major target pest, cotton bollworm (Helicoverpa armigera), in northern China. Laboratory bioassay results show that susceptibility to Cry1Ac was significantly lower in 13 field populations from northern China, where Bt cotton has been planted intensively, than in two populations from sites in northwestern China where exposure to Bt cotton has been limited. Susceptibility to Bt toxin Cry2Ab did not differ between northern and northwestern China, demonstrating that resistance to Cry1Ac did not cause cross-resistance to Cry2Ab, and implying that resistance to Cry1Ac in northern China is a specific adaptation caused by exposure to this toxin in Bt cotton. Despite the resistance detected in laboratory bioassays, control failures of Bt cotton have not been reported in China. This early warning may spur proactive countermeasures, including a switch to transgenic cotton producing two or more toxins distinct from Cry1A toxins.


The influence of the carbonic anhydrase inhibitor, benzolamide (CL-11,366), on the reabsorption of chloride, sodium, and bicarbonate in the proximal tubule of the rat.
Benzolamide (CL-11,366), a potent carbonic anhydrase inhibitor, was used to examine the influence of carbonic anhydrase inhibition on the reabsorption of chloride, sodium, and bicarbonate in the rat proximal convoluted tubule. Administration of 2 mg/kg benzolamide was associated with a decrease in the tubular fluid/plasma (TF/P) chloride ratio from 1.19+/-0.10 (SEM) to 1.06+/-0.01, and an increase in the TF/P bicarbonate ratio from 0.181+/-0.02 to 0.584+/-0.02. This dose of benzolamide significantly reduced proximal fractional reabsorption of chloride by 29.14%, of sodium by 34.3%, and of bicarbonate by 35.64%. These results indicate that benzolamide administration inhibits the reabsorption of all three electrolytes in the proximal convoluted tubule. Although 20 mg/kg benzolamide accentuated the changes in fractional reabsorption, the differences between 2 and 20 mg/kg were not statistically significant. Inhibition of proximal tubular cytoplasmic and luminal carbonic anhydrase could well explain the diminished bicarbonate reabsorption and a fraction of the diminished sodium reabsorption noted in these studies. The fall in chloride reabsorption, and a portion of the fall in sodium reabsorption, however, may be a direct or indirect consequence of carbonic anhydrase inhibition or of an influence of benzolamide on a transport mechanism not dependent upon carbonic anhydrase.


Prevalence and predictors of Chlamydia trachomatis and Neisseria gonorrhoeae among HIV-negative pregnant women in Kenya.
Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infections in pregnancy contribute to adverse perinatal outcomes. We identified predictors of CT and/or NG infection among pregnant Kenyan women. Women without HIV were enrolled at two antenatal clinics in Western Kenya. CT and NG were assessed using endocervical samples for nucleic acid amplification tests (NAAT). Poisson regression models were used to evaluate potential CT/NG risk factors. Classification and regression trees (CART) were generated to evaluate the joint effects of predictors. Overall, 1276 women had both CT and NG assessments. Women enrolled at a median of 26 weeks gestation (IQR 22-31), median age was 22 years (IQR 19-27), and 78% were married. In total, 98 (7.7%) tested positive for CT/NG: 70 (5.5%) for CT, 32 (2.5%) for NG, 4 of whom (0.3%) had co-infections. Two-thirds (66%) of CT/NG cases were asymptomatic and would have been missed with only syndromic management. Risk factors of CT/NG included age < 22 years, crowded living conditions, being unmarried, being in partnerships for <1-year, abnormal vaginal discharge, previous STI history, and T. vaginalis diagnosis(p < 0.05). CART analyses identified unmarried women <22 years in relationships for <1 year as 6.1 times more likely to have CT/NG compared to women without these characteristics (26% vs. 6%, adjusted prevalence ratio = 6.1, 95% CI: 3.55-10.39, p < 0.001). CT/NG was frequently asymptomatic and common among young unmarried women in newer partnerships in this cohort. Integrating CT/NG testing into routine antenatal care may be beneficial, especially for young women in Kenya.


Posttraumatic stress disorder: an exploratory study examining rates of trauma and PTSD and its effect on client outcomes in community mental health.
Rates of trauma and Posttraumatic Stress Disorder (PTSD) were examined in order to compare the profile in clients of an Australian Public Mental Health Service with that reported in the international literature for clients with major mental illness and to explore the effect of this on client health outcomes. Potential factors contributing to increased levels of trauma/PTSD in this group of clients and the issue of causality between PTSD and subsequent mental illness was also explored. A convenience sample of 29 clients was screened for trauma and PTSD using the Posttraumatic Stress Diagnostic Scale (PDS) and selected outcome measures. Paired and independent samples t-test and ANOVA were applied to the data. High levels of undocumented trauma and PTSD were found. Twenty clients, (74%) reported exposure to multiple traumatic events; 33.3% (9) met DSM IV diagnostic criteria for PTSD. Significant difference was found for PTSD symptomatology, severity and impairment and for client and clinician-rated scores of Quality of Life (QOL) outcomes in the PTSD group. No effect for PTSD symptomatology on the Working Alliance (WA) was found. Factors that may influence higher rates of PTSD in this group were identified and included issues associated with the population studied, the predominance of assaultive violence found, and vulnerability and risks factors associated with re-traumatisation within the social and treating environments. A similar trauma and PTSD profile to that reported in the international literature, including greater levels of trauma and PTSD and a poorer QOL, was found in this small sample of clients. It is postulated that the increased levels of trauma/PTSD as reported for persons with major mental illness, including those found in the current study, are primarily related to the characteristics of the population that access public mainstream psychiatric services and that these factors have specific implications for service delivery, and raise issues of efficiency and effectiveness of resource use in achieving successful outcomes in public mental health services for clients with co-morbid PTSD. Further research with a more rigorous design is needed to test these preliminary findings within Australian Community Mental Health Services.


Liraglutide treatment for the prevention of glucose tolerance deterioration in women with prior gestational diabetes mellitus: A 52-week randomized controlled clinical trial.
We investigated the effect of 52-week treatment with liraglutide, a glucagon-like peptide 1 receptor agonist, on glucose tolerance and incretin effect in women with previous gestational diabetes mellitus (pGDM). Women with overweight/obesity and pGDM were randomized to once daily subcutaneous liraglutide 1.8 mg or placebo for 52 weeks. Participants underwent oral glucose tolerance test (OGTT) and isoglycaemic intravenous glucose infusion at baseline and at 52 weeks, and an additional OGTT after the drug wash-out. In total, 104 women [age: mean ± SD, 38 ± 5 years; fasting plasma glucose (FPG): 5.5 ± 0.4 mmol/L; glycated haemoglobin (HbA1c): 33 ± 4 mmol/mol, bodyweight: 88.2 ± 14.8 kg, body mass index: 31.1 ± 4.3 kg/m2 ] were assigned to liraglutide (n = 49) or placebo (n = 55). Estimated treatment difference (ETD) for area under curve during OGTT was -173 (95% confidence interval -250 to -97) mmol/L × min, p < .0001, but after wash-out the difference disappeared [ETD 58 (-30 to 146) mmol/L × min, p = .536]. Liraglutide reduced FPG [ETD -0.2 (-0.4 to -0.1) mmol/L, p = .018], HbA1c [-2.2 (-3.5 to -0.8) mmol/mol, p = .018] and bodyweight [-3.9 (-6.2 to -1.6) kg, p = .012]. No change in the incretin effect was observed. The number of women with prediabetes was reduced from 64% to 10% with liraglutide vs. 50% with placebo [adjusted odds ratio 0.10 (0.03-0.32), p = .002]. Treatment with liraglutide for 52 weeks improved glucose tolerance, FPG, HbA1c and bodyweight in women with overweight/obesity and pGDM. Progression to prediabetes while on drug was markedly reduced, but after a 1-week drug wash-out, the effect was lost.


Reconstructed human skin produced in vitro and grafted on athymic mice.
The best alternative to a split-thickness graft for the wound coverage of patients with extensive burns should be in vitro reconstructed autologous skin made of both dermis and epidermis and devoid of exogenous extracellular matrix proteins and synthetic material. We have designed such a reconstructed human skin (rHS) and present here its first in vivo grafting on athymic mice. The rHS was made by culturing newborn or adult keratinocytes on superimposed fibrous sheets obtained after culturing human fibroblasts with ascorbic acid. Ten days after keratinocyte seeding, reconstructed skins were either cultured at the air-liquid interface or grafted on athymic mice. We present the macroscopic, histologic, and phenotypic properties of such tissues in vitro and in vivo after grafting on nude mice. After maturation in vitro, the reconstructed skin exhibited a well-developed human epidermis that expressed differentiated markers and basement membrane proteins. Four days after grafting, a complete take of all grafts was obtained. Histological analysis revealed that the newly generated epidermis of newborn rHS was thicker than that of adult rHS after 4 days but similar 21 days after grafting. The basement membrane components (bullous pemphigoid antigens, laminin, and type IV and VII collagens) were detected at the dermo-epidermal junction, showing a continuous line 4 days after grafting. Ultrastructural studies revealed that the basement membrane was continuous and well organized 21 days after transplantation. The macroscopic aspect of the reconstructed skin revealed a resistant, supple, and elastic tissue. Elastin staining and elastic fibers were detected as a complex network in the rHS that contributes to the good elasticity of this new reconstructed tissue. This new rHS model gives supple and easy to handle skins while demonstrating an adequate wound healing on mice. These results are promising for the development of this skin substitute for permanent coverage of burn wounds.


Assessing response, remission, and treatment resistance in patients with obsessive-compulsive disorder with and without tic disorders: results from a multicenter study.
Highlighting the relationship between obsessive-compulsive disorder (OCD) and tic disorder (TD), two highly disabling, comorbid, and difficult-to-treat conditions, Diagnostic and Statistical Manual of Mental Disorders, fifth edition (DSM-5) acknowledged a new "tic-related" specifier for OCD, ie, obsessive-compulsive tic-related disorder (OCTD). As patients with OCTD may frequently show poor treatment response, the aim of this multicenter study was to investigate rates and clinical correlates of response, remission, and treatment resistance in a large multicenter sample of OCD patients with versus without tics. A sample of 398 patients with a DSM-5 diagnosis of OCD with and without comorbid TD was assessed from 10 different psychiatric departments across Italy. For the purpose of the study, treatment response profiles in the whole sample were analyzed comparing the rates of response, remission, and treatment-resistance as well as related clinical features. Multivariate logistic regressions were performed to identify possible factors associated with treatment response. The remission group was associated with later ages of onset of TD and OCD. Moreover, significantly higher rates of psychiatric comorbidities, TD, and lifetime suicidal ideation and attempts emerged in the treatment-resistant group, with larger degrees of perceived worsened quality of life and family involvement. Although remission was associated with later ages of OCD and TD onset, specific clinical factors, such as early onset and presence of psychiatric comorbidities and concomitant TD, predicted a worse treatment response with a significant impairment in quality of life for both patients and their caregivers, suggesting a worse profile of treatment response for patients with OCTD.


Tendon end separation with loading in an Achilles tendon repair model: comparison of non-absorbable vs. absorbable sutures.
Rupture of the Achilles tendon often leads to long-term morbidity, particularly calf weakness associated with tendon elongation. Operative repair of Achilles tendon ruptures leads to reduced tendon elongation. Tendon lengthening is a key problem in the restoration of function following Achilles tendon rupture. A study was performed to determine differences in initial separation, strength and failure characteristics of differing sutures and numbers of core strands in a percutaneous Achilles tendon repair model in response to initial loading. Nineteen bovine Achilles tendons were repaired using a percutaneous/minimally invasive technique with a combination of a modified Bunnell suture proximally and a Kessler suture distally, using non-absorbable 4-strand 6-strand repairs and absorbable 8-strand sutures. Specimens were then cyclically loaded using phases of 10 cycles of 100 N, 100 cycles of 100 N, 100 cycles of 190 N consistent with early range of motion training and weight-bearing, before being loaded to failure. Pre-conditioning of 10 cycles of 100 N resulted in separations of 4 mm for 6-strand, 5.9 mm for 4-strand, but 11.5 mm in 8-strand repairs, this comprised 48.5, 68.6 and 72.7% of the separation that occurred after 100 cycles of 100 N. The tendon separation after the third phase of 100 cycles of 190 N was 17.4 mm for 4-strand repairs, 16.6 mm for 6-strand repairs and 26.6 mm for 8-strand repairs. There were significant differences between the groups (p < 0.0001). Four and six strand non-absorbable repairs had significantly less separation than 8-strand absorbable repairs (p = 0.017 and p = 0.04 respectively). The mean (SEM) ultimate tensile strengths were 4-strand 464.8 N (27.4), 6-strand 543.5 N (49.6) and 8-strand 422.1 N (80.5). Regression analysis reveals no significant difference between the overall strength of the 3 repair models (p = 0.32) (4 vs. 6: p = 0.30, 4 vs. 8: p = 0.87; 6 vs. 8: p = 0.39). The most common mode of failure was pull out of the Kessler suture from the distal stump in 41.7% of specimens. The use of a non-absorbable suture resulted in less end-to-end separation when compared to absorbable sutures when an Achilles tendon repair model was subject to cyclical loading. Ultimate failure occurred more commonly at the distal Kessler suture end although this occurred with separations in excess of clinical failure. The effect of early movement and loading on the Achilles tendon is not fully understood and requires more research.


CON: The hypotension prediction index is a validated predictor of hypotension.
The Hypotension Prediction Index (HPI) algorithm is a commercial prediction algorithm developed to predict hypotension, a mean arterial pressure (MAP) below 65 mmHg. Although HPI has been investigated in several studies, recent concerns of have been raised regarding HPI's predictive abilities, which may have been overstated. A selection bias may have forced the HPI algorithm to learn almost exclusively from MAP. This CON position paper describes the selection bias further and summarises the scientific status of HPI's predictive abilities, including the meaning of a recent erratum retracting the primary conclusion of a published HPI validation study. We argue that the HPI algorithm needs re-validation or complete re-development to achieve a clinically relevant 'added value' in comparison with the predictive performance of a simple and costless MAP alarm threshold in the range of 70 to 75 mmHg.


16 alpha-hydroxylation of estradiol: a possible risk marker for breast cancer.
From these results we may conclude that estradiol 16 alpha-hydroxylation is highly correlated with tumor incidence, and that the reaction is partly regulated by MMTV and the rest by genetic influences. Elevated hydroxylation appears to be an autosomal dominant trait that is highly specific for estradiol. It is also pertinent that the product of the 16 alpha-hydroxyestrone reaction is a potent estrogen that is capable of binding covalently to amino acids and nucleotides, including the estrogen receptor molecule. The results obtained in these studies establish the usefulness of the mouse model for studying the interrelationship between enhanced 16-hydroxylation of estradiol and the incidence of mammary tumors.


Effects of polyamines on protein synthesis and growth of Escherichia coli.
The polyamines (PA) putrescine, spermidine, and spermine have numerous roles in the growth of both prokaryotic and eukaryotic cells. For example, it is well known that putrescine and spermidine are strongly involved in proliferation and viability of Escherichia coli cells. Studies of polyamine functions and distributions in E. coli cells have revealed that polyamines mainly exist as an RNA-polyamine complex. Polyamines stimulate the assembly of 30S ribosomal subunits and thereby increase general protein synthesis 1.5- to 2.0-fold. Moreover, these studies have shown that polyamines stimulate synthesis of 20 different proteins at the level of translation, which are strongly involved in cell growth and viability. The genes encoding these 20 different proteins were termed as the "polyamine modulon." We here review the mechanism of activation of 30S ribosomal subunits and stimulation of specific proteins. Other functions of polyamines in E. coli are also described.


Experiential cognitive therapy in the treatment of panic disorders with agoraphobia: a controlled study.
The use of a multicomponent cognitive-behavioral treatment strategy for panic disorder with agoraphobia is actually one of the preferred therapeutic approaches for this disturbance. This method involves a mixture of cognitive and behavioral techniques that are intended to help patients identify and modify their dysfunctional anxiety-related thoughts, beliefs and behavior. The paper presents a new treatment protocol for Panic Disorder and Agoraphobia, named Experiential-Cognitive Therapy (ECT) that integrates the use of virtual reality (VR) in a multicomponent cognitive-behavioral treatment strategy. The VR software used for the trial is freely downloadable: www.cyberpsychology.info/try.htm. Moreover, the paper presents the result of a controlled study involving 12 consecutive patients aged 35-53. The selected subjects were randomly divided in three groups: ECT group, that experienced the Cognitive Behavioral Therapy-Virtual Reality assisted treatment (eight sessions), a CBT group that experienced the traditional Cognitive Behavioral approach (12 sessions) and a waiting list control group. The data showed that both CBT and ECT could significantly reduce the number of panic attacks, the level of depression and both state and trait anxiety. However, ECT procured these results using 33% fewer sessions than CBT. This datum suggests that ECT could be better than CBT in relation to the "cost of administration," justifying the added use of VR equipment in the treatment of panic disorders.