Effects of concanavalin A, phytohemagglutinin, pokeweed mitogen, and lipopolysaccharide on the replication and immunoglobulin synthesis by canine peripheral blood lymphocytes in vitro.
Canine peripheral blood lymphocytes (cPBLs) were used to investigate the mitogenic effects of Con A (concanavalin A), LPS (lipopolysaccharide), PHA (phytohemagglutinin), and PWM (pokeweed mitogen) in vitro by measuring tritium-labeled thymidine [( 3H]thymidine) incorporation and immunoglobulin (Ig) secretion. An ELISA specific for canine IgG and IgM showed that cPBLs secreted significantly more IgG than IgM in response to mitogen concentrations from 30,000 to 0.03 ng/10(5) cells. The optimal stimulating dose of mitogen for lymphocyte response measured by IgG secretion was over a much narrower range of concentration than was the [3H]thymidine incorporation measured response. At a concanavalin A dose where there was increased [3H]thymidine incorporation with a decrease in IgG secretion, it appeared that an active suppression of the IgG response was induced.


Field-flow fractionation and biotechnology.
The gentle separation mechanism has made field-flow fractionation particularly suited to samples of biotechnological interest, from proteins and nucleic acids to viruses, subcellular units and whole cells. Recent progress in field-flow fractionation technology, as well as the development of coupled techniques combining field-flow fractionation capabilities with the specificity and sensitivity of well-established analytical methods, opens up new biotechnological applications for field-flow fractionation. The most recent appealing applications include: sorting and fingerprinting of bacteria for whole-cell vaccine production; noninvasive and tagless sorting of immature and stem cells; separation of intact proteins and enzymes in top-down proteomics; and the development of flow-assisted, multianalyte immunoassays using nano- and micron-sized particles with immobilized biomolecules.


An absorbance-based micro-fluidic sensor for diffusion coefficient and molar mass determinations.
The H-Sensor reported herein is a micro-fluidic device compatible with flow injection analysis (FIA) and high performance liquid chromatography (HPLC). The device detects analytes at two separate off-chip absorbance flow cells, providing two simultaneous absorbance measurements. The ratio of these two absorbance signals contains analyte diffusion coefficient information. A theoretical model for the sensing mechanism is presented. The model relates the signal Ratio to analyte diffusion coefficient. The model is qualitatively evaluated by comparing theoretical and experimental signal Ratio values. Experimental signal Ratios were collected via FIA for a variety of analytes, including sodium azide, benzoic acid, amino acids, peptides, and proteins. Measuring absorbance at multiple wavelengths provides higher order data allowing the analyte signals from mixtures to be deconvolved via classical least squares (CLS). As a result of the H-Sensor providing two simultaneous signals as a function of time for each sample injection, two simulated second-order HPLC chromatograms were generated using experimental H-Sensor data. The chemometric deconvolution method referred to as the generalized rank annihilation method (GRAM) was used to demonstrate chromatographic and spectroscopic deconvolution. GRAM also provides the signal Ratio value, therefore simultaneously obtaining the analyte diffusion coefficient information during deconvolution. The two chromatograms successfully serve as the standard and unknown for the GRAM deconvolution. GRAM was evaluated on chromatograms at various chromatographic resolutions. GRAM was found to function to a chromatographic resolution at and above 0.25 with a percent quantitative error of less then 10%.


[Effect of pentamine on vascular tissue metabolism in experimental vascular pathology].
It has been demonstrated in experiments on cats and rabbits that a single and chronic administration of pentamine (1 mg/kg) produced a distinct variation in carbohydrate-energy metabolism in the tissue of different blood vessels, enhancing the glycolytic formation of ATP. Chronic administration of pentamine corrected metabolic disorders in rabbit vascular tissue in experimental pituitrin-induced hypertension.


Using Teach-Back to Understand Participant Behavioral Self-Monitoring Skills Across Health Literacy Level and Behavioral Condition.
To assess differences, by health literacy status and behavioral condition, in participants' abilities to self-monitor behaviors accurately and recall key behavioral messages using data from a teach-back call. Cross-sectional. Rural, southwestern Virginia. Adults (n = 301). The majority were female (81.1%), 31.9% had a high school education or less, 66.1% earned < $25,000/y, and 32.9% were low health literate. First class session of 2 community-based behavioral interventions: SIPsmartER (reduce sugar-sweetened beverage intake) or MoveMore (increase physical activity). Reported accuracy of behavioral diary completion, proportion of behavioral messages recalled during the first round of teach-back, and rounds of teach-back. Descriptive statistics and generalized linear model. Low health literate participants were significantly less accurate in diary completion (P < .001), recalled fewer behavioral messages correctly (P < .001), and needed more rounds of teach-back (P < .001) than high health literate participants. Compared with SIPsmartER participants, MoveMore participants more accurately completed diaries (P = .001) but recalled a lower proportion of behavioral messages correctly (P < .001) and required more rounds of teach-back (P < .001). Health literacy status and behavioral target affect the ability to self-monitor and recall key concepts. Researchers should consider using teach-back early in the intervention to assess and reinforce participants' ability to self-monitor.


Dietary interventions affecting chylomicron and chylomicron remnant clearance.
Interest in effects of diet on postprandial lipoproteins has increased in recent years as a result of accumulating evidence for adverse cardiovascular consequences of elevated concentrations of triglyceride rich lipoproteins. Particular attention has been given to ability of different fatty acids to modulate postprandial lipoprotein responses because of evidence for both harmful and protective cardiovascular properties of the saturated, monounsaturated and omega-6 and omega-3 polyunsaturated fatty acid (PUFA) classes. Evidence for direct atherogenic properties of chylomicron remnants has led to attempts to monitor effects of diet specifically on this lipoprotein class. Limitations in the methods employed to measure chylomicron remnants and the small number of human studies which have evaluated effects of meal, and background diet, fatty acid composition, makes it difficult to draw definitive conclusions at the present time. However consideration of data from both animal and human studies tends to support the conclusion that diets, and meals, rich in PUFA (particularly long chain omega-3 PUFA), result in attenuated postprandial responses of the intestinally-derived lipoproteins. Attenuated responses to high PUFA meals appear to be due to greater rates of clearance and greater activation of lipoprotein lipase (LPL). Attenuated responses to high PUFA background diets may be due to adaptive changes involving both accelerated rates of clearance in peripheral tissues and liver, as well as decreased output of the competitor for chylomicron clearance, very low density lipoprotein (VLDL).


Optical spectroscopy of individual objects.
With the introduction of single-molecule spectroscopy it was immediately realised that studying molecules and molecular aggregates individually opens up a completely new approach to elucidate information commonly washed out by the ensemble average process. In contrast with ensemble experiments, this technique permits us to determine the whole distribution of parameters rather than their moments. This allows a more direct verification of theoretical models without the need to average over (unknown) distributions to calculate observables. Here we illustrate how single-molecule spectroscopy can be utilised to circumvent the classical diffraction limit of microscopy and show an investigation of the electronic structure of pigment protein complexes that are relevant for bacterial photosynthesis.


Neuronal membrane depolarization and the control of cholinergic muscarinic receptors: selective effect on different neuronal cell types.
1. The possibility that a long-lasting neuronal activation regulates the expression of muscarinic cholinergic receptors was studied with three cultured neuronal cell lines. 2. Continuous depolarization of a subclone of the neuroblastoma-glioma NG108-15 hybrid cells with potassium chloride increased by 45-75% the number of cholinergic muscarinic receptors, monitored with 3H-QNB, whereas a short incubation with KCl for 10 min or 6 hr had no effect. 3. The calcium channel blocker verapamil increased the effect of KCl. 4. Two cell lines, named SC9 and WC5, that originate from the rat brain, also bind 3H-QNB. They were therefore used to test whether the effect of chronic depolarization is universal. Depolarized SC9 and WC5 cells, in the presence or absence of verapamil, did not show an increased 3H-QNB binding. 5. Muscarinic receptors of both SC9 and WC5 cells have a higher affinity to pirenzepine than the M-3 receptor subtype of the neuroblastoma-glioma cells, suggesting therefore that the two rat brain cell lines possess M-1 or M-2 receptors. 6. The physiological significance of this differential role of depolarization on the expression of different muscarinic receptors is discussed in the context of their postreceptor second messengers.


Vascular permeability changes involved in tumor metastasis.
Cancer cell extravasation resembles the leukocyte recruitment during inflammation. Evidence suggests that cancer cells need to weaken the interendothelial junctions in order to cross the endothelial barrier. Several tumor-derived vasoactive compounds have been pointed out to drive this increase in vascular permeability: VEGF, Angptl4, CCL2, SDF-1, etc. Therefore, tumor cells have a wide repertoire of soluble factors to increase vascular permeability in order to colonize new tissues. Tumor soluble factors activate different signaling pathways to induce interendothelial junction disassembly, one common element is Src kinase. Here we summarize the relevant current knowledge about vascular permeability changes involved in tumor metastasis.


[Growth of cultured blood cells in the presence of the cryoprotector polyethylene oxide 400].
The effect of polyethylene oxide 400 (PEO-400) cryoprotector on the mitotic activity of human peripheral blood leukocytes stimulated with PHA was studied in a culture. The cytologic peculiarities of the blood cell growth on addition of PEO-400 prior to cultivation are manifested in the preservation of the transformation capacity and mitosis despite a significant reduction of the mitotic activity in comparison with control cultures.


Capturing intermediate filament networks.
Mapping intermediate filaments in three dimensions reveals that the organization of these filaments differs across cell types.


Impact of magnetic resonance imaging versus CT on nasopharyngeal carcinoma: primary tumor target delineation for radiotherapy.
Our aim was to assess the capacity of CT versus MRI for delineating to the primary tumor extent of nasopharyngeal carcinoma (NPC) in treated patients. From December 1997 to April 2000, 258 patients with NPC were enrolled. We focused on the primary tumor extension and the discrepancy between CT and MRI. The delineation of tumor invasion was crucial for determination of the gross tumor volume (GTV) before radiation therapy. A total of 104 patients (40.3%) had intracranial infiltration detected by MRI, whereas CT showed negative findings (p = 6.879 x 10(-11)). Once the pterygopalatine fossa was involved, the chance of intracranial invasion was increased (96.1%). The detectable percentage of pterygopalatine fossa involvement accompanying intracranial invasion was higher with MRI than with CT (96.1% vs 56.9%). More detailed information about T and N classification of NPC was provided by MRI than by CT, which led to better target delineation for radiotherapy.


Dementia Diagnoses and Treatment in Geriatric Ward Patients: A Cross-Sectional Study in Poland.
The study aimed to determine the prevalence of dementia, its types, and treatment in geriatric ward patients. A cross-sectional study of 406 patients (77.8% women, median age 82, IQR (77-86) years) who underwent a comprehensive geriatric assessment in one of the Polish hospitals between September 2014 and April 2015 was conducted. Dementia was diagnosed in 132 (32.5%) patients (46% mixed dementia; 32% Alzheimer's disease; 10%vascular dementia; 5% dementia in Parkinson's disease; 4% frontotemporal; 3% atypical parkinsonism). A total of 95 (72%) dementia cases were not detected before, and in the above half of these patients, it was not mentioned in the referral document. Only 33.3% of dementia patients were on cognitive enhancers (donepezil, rivastigmine, or memantine); 36.4% received antipsychotics, 45.5% received anti-depressants, 25.8% received nootropics, and 16.7% received anxiolytics/hypnotics. The results confirmed the high incidence of underdiagnoses and undertreatment of dementia in patients admitted to the geriatric ward. It is partly due to the lack of systematic cognitive assessment in primary care settings, although other factors can play a role.


Bag-mask ventilation and direct laryngoscopy versus intubating laryngeal mask airway: a manikin study of hands-on times during cardiopulmonary resuscitation.
The percentage of hands-on time during cardiopulmonary resuscitation is a major determinant of patient outcome. We hypothesized that airway management with the intubating laryngeal mask airway (ILMA) would give greater hands-on time than with bag-mask ventilation (BMV), followed by direct laryngoscopy (DL), particularly in difficult-to-manage airways. Thirty paramedics and 40 medical students performed four standardized, 6-min cardiopulmonary resuscitation scenarios with the SimMan3G in a random sequence. These were normal and difficult-to-manage airways using either BMV+DL or ILMA. The time to the first successful ventilation was significantly longer with the ILMA (P<0.001). Hands-on time was lower for the ILMA after 2 min (67±8 vs. 81±8 s for BMV+DL, P<0.001), but was then significantly greater from the third minute onward (115±11 vs. 104±9 s for BMV+DL, P<0.001). The success rate of the first intubation attempt was higher and the time to ET placement was shorter with the ILMA, especially in the difficult-to-manage airway (P<0.001). In this manikin-based study, hands-on time was greater with the ILMA than with BMV+DL. The ILMA was particularly useful in increasing hands-on times in the difficult-to-manage airway.


Molecular mechanisms of nitric oxide-induced growth arrest and apoptosis in fetal pulmonary arterial smooth muscle cells.
Superoxide plays an important role in pulmonary arterial smooth muscle cell (SMC) proliferation and survival. The rapid reaction between superoxide and nitric oxide (NO) to form peroxynitrite suggests that endothelium-derived NO may influence adjacent SMC growth. To investigate this possibility, we determined the dose-dependent effects of NO on the proliferation and viability of pulmonary arterial SMC isolated from fetal lambs (FPASMC). Using fluorescence microscopy we found a dose-dependent decrease in superoxide levels in FPASMC treated with the NO donor spermine NONOate. This was associated with an increase in peroxynitrite-mediated protein nitration. At doses between 50 and 250 microM, spermine NONOate attenuated serum-induced FPASMC proliferation resulting in a G(0)/G(1) cell cycle arrest. This process involved a decrease in levels of cyclin A and an increase in the nuclear localization of p21 and p27. Furthermore, 500 microM spermine NONOate decreased viable cell number by inducing programmed cell death: FPASMC treated with 500 microM spermine NONOate displayed a loss of mitochondrial membrane potential, followed by caspase activation and DNA fragmentation. These data suggest that NO inhibits superoxide-induced proliferation of FPASMC and at higher doses induces apoptosis. NO donors may therefore prove to be useful therapeutic tools to treat diseases resulting from excessive proliferation of vascular smooth muscle.


Capacity of rat liver for pyrimidine synthesis and catabolism during fetal and neonatal development.
The capacity of minces of rat liver to synthesize and degrade pyrimidines during fetal and neonatal development was examined. Pyrimidine synthesis was determined by measuring the rate of incorporation of NaH14CO3 into orotic acid. Pyrimidine catabolism was estimated by measuring the generation of 14CO2 from [2-14C]uridine. The incorporation of [2-14C]uridine into RNA was determined simultaneously with measurements of uridine catabolism. The activity of beta-ureidopropionase, the enzyme which catalyzes the terminal reaction in the dihydropyrimidine catabolic pathway, was also monitored in cell-free extracts of liver throughout the perinatal period. Catabolic activity was detected at the earliest stage of gestation examined (16 days) and rose sharply during fetal development to reach adult levels at birth or shortly thereafter. A similar rise in the activity of beta-ureidopropionase was somewhat delayed when compared with the rise in overall catabolic activity; the enzyme activity at birth was about half the adult level. By way of contrast, the incorporation of NaH14CO3 into orotic acid and [2-14C]uridine into RNA were highest in 16-day fetal liver and declined sharply with fetal and neonatal development. These results demonstrate an appreciable capacity for pyrimidine catabolism in fetal liver, and also contribute to growing evidence that fetal tissues are capable of meeting their pyrimidine requirements through de novo synthesis. The contrast observed between the rate of synthesis of orotic acid and the capacity for pyrimidine degradation throughout perinatal development fits the pattern which has emerged from other studies showing the pathways for the anabolism and catabolism of pyrimidines to be regulated inversely to one another.


Genetic engineering of Enterobacter aerogenes with the Vitreoscilla hemoglobin gene: cell growth, survival, and antioxidant enzyme status under oxidative stress.
Hemoglobins in unicellular organisms, like the one here in the bacterium Vitreoscilla, have greater chemical reactivity than their homologues in multicellular organisms. They can catalyze redox reactions and may protect cells against oxidative stress. The ability of Vitreoscilla hemoglobin to complement deficiencies of terminal cytochrome oxidases in Escherichia coli also suggests that this hemoglobin can receive electrons during respiration. In this study, a recombinant strain of Enterobacter aerogenes engineered to produce the Vitreoscilla Hb was investigated with regard to its susceptibility to oxidative stress. The culture response to oxidative stress produced by exogenously applied hydrogen peroxide was characterized in terms of cell growth, survival and the activities of two key antioxidant enzymes (catalase and superoxide dismutase). The influence of the physiological state of the cells and different media upon these culture dynamics was determined. Results showed that the hemoglobin-expressing strain is quite distinct in terms of growth/survival properties and activity of antioxidant enzymes from that of non-hemoglobin counterparts.


The influence of the depth of embryo replacement into the uterine cavity on implantation rates after IVF: a controlled, ultrasound-guided study.
Traditionally, embryo transfer after IVF has been performed blindly and placing the embryos approximately 1 cm below the fundal endometrial surface. However, it has been suggested that transferring embryos rather lower in the uterine cavity or high in the uterus may improve implantation rates. Nevertheless, there has not yet been a controlled trial to prove this theory. This prospective randomized study investigates the influence of the depth of embryo replacement on the implantation rate after embryo transfer carried out under transabdominal ultrasound guidance. A total of 180 consecutive patients undergoing ultrasound-guided embryo transfer were randomized to three study groups according to the distance between the tip of the catheter and the uterine fundus at the moment of the embryo deposition in the lumen of the endometrial cavity: group 1: 10 +/- 1.5 mm; group 2: 15 +/- 1.5 mm; group 3: 20 +/- 1.5 mm. There was equal distribution between all three study groups regarding the main demographic and baseline characteristics of the patients, ovarian response, oocyte retrieval and IVF outcome, as well as the characteristics of embryo transfer and luteal phase support. The position of the catheter tip in relation to the fundal endometrial surface in groups 1 (10.2 +/- 0.9 mm), 2 (14.6 +/- 0.7 mm) and 3 (19.3 +/- 0.8 mm) was significantly different. Implantation rate was significantly higher (P < 0.05) in groups 2 (31.3%) and 3 (33.3%) compared with group 1 (20.6%). The depth of the embryo replacement into the uterine cavity may influence implantation rates, and thus it should be considered as an additional procedure among factors recently proposed as associated with successful embryo transfer after IVF.


Epistatic participation of REV1 and REV3 in the formation of UV-induced frameshift mutations in cell cycle-arrested yeast cells.
Mutations arising in times of cell cycle arrest may provide a selective advantage for unicellular organisms adapting to environmental changes. For multicellular organisms, however, they may pose a serious threat, in that such mutations in somatic cells contribute to carcinogenesis and ageing. The budding yeast Saccharomyces cerevisiae presents a convenient model system for studying the incidence and the mechanisms of stationary-phase mutation in a eukaryotic organism. Having studied the emergence of frameshift mutants after several days of starvation-induced cell cycle arrest, we previously reported that all (potentially error-prone) translesion synthesis (TLS) enzymes identified in S. cerevisiae did not contribute to the basal level of spontaneous stationary-phase mutations. However, we observed that an increased frequency of stationary-phase frameshift mutations, brought about by a defective nucleotide excision repair (NER) pathway or by UV irradiation, was dependent on Rev3p, the catalytic subunit of the TLS polymerase zeta (Pol zeta). Employing the same two conditions, we now examined the effect of deletions of the genes coding for polymerase eta (Pol eta) (RAD30) and Rev1p (REV1). In a NER-deficient strain background, the increased incidence of stationary-phase mutations was only moderately influenced by a lack of Pol eta but completely reduced to wild type level by a knockout of the REV1 gene. UV-induced stationary-phase mutations were abundant in wild type and rad30Delta strains, but substantially reduced in a rev1Delta as well as a rev3Delta strain. The similarity of the rev1Delta and the rev3Delta phenotype and an epistatic relationship evident from experiments with a double-deficient strain suggests a participation of Rev1p and Rev3p in the same mutagenic pathway. Based on these results, we propose that the response of cell cycle-arrested cells to an excess of exo- or endogenously induced DNA damage includes a novel replication-independent cooperative function of Rev1p and Pol zeta, which has the potential to generate mutations.


Generation of fully human anti-GPC3 antibodies with high-affinity recognition of GPC3 positive tumors.
The acceleration of therapeutic antibody development has been motivated by the benefit to and their demand for human health. In particular, humanized transgenic antibody discovery platforms, combined with immunization, hybridoma fusion and/or single cell DNA sequencing are the most reliable and rapid methods for mining the human monoclonal antibodies. Human GPC3 protein is an oncofetal antigen, and it is highly expressed in most hepatocellular carcinomas and some types of squamous cell carcinomas. Currently, no fully human anti-GPC3 therapeutic antibodies have been reported and evaluated in extensive tumor tissues. Here, we utilized a new humanized transgenic mouse antibody discovery platform (CAMouse) that contains large V(D)J -regions and human gamma-constant regions of human immunoglobulin in authentic configurations to generate fully human anti-GPC3 antibodies. Our experiments resulted in four anti-GPC3 antibodies with high-specific binding and cytotoxicity to GPC3 positive cancer cells, and the antibody affinities are in the nanomolar range. Immunohistochemistry analysis demonstrated that these antibodies can recognize GPC3 protein on many types of solid tumors. In summary, the human anti-human GPC3 monoclonal antibodies described here are leading candidates for further preclinical studies of cancer therapy, further, the CAMouse platform is a robust tool for human therapeutic antibody discovery.


Croquemort elicits activation of the immune deficiency pathway in ticks.
The immune deficiency (IMD) pathway directs host defense in arthropods upon bacterial infection. In Pancrustacea, peptidoglycan recognition proteins sense microbial moieties and initiate nuclear factor-κB-driven immune responses. Proteins that elicit the IMD pathway in non-insect arthropods remain elusive. Here, we show that an Ixodes scapularis homolog of croquemort (Crq), a CD36-like protein, promotes activation of the tick IMD pathway. Crq exhibits plasma membrane localization and binds the lipid agonist 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol. Crq regulates the IMD and jun N-terminal kinase signaling cascades and limits the acquisition of the Lyme disease spirochete B. burgdorferi. Additionally, nymphs silenced for crq display impaired feeding and delayed molting to adulthood due to a deficiency in ecdysteroid synthesis. Collectively, we establish a distinct mechanism for arthropod immunity outside of insects and crustaceans.


Effect of dipyridamole on spontaneous platelet aggregation in whole blood decreases with the time after venepuncture: evidence for the role of ADP.
Spontaneous platelet aggregation (SPA) was studied in human whole blood at 3, 5, 10, 20, 30, 40, and 60 minutes after venepuncture. Using a whole blood platelet counter, SPA was quantified by measuring the fall in single platelet count upon rollermixing aliquots of citrated blood at 37 degrees C. The extent of SPA increased with the time after venepuncture, with a correlation coefficient of 0.819. The inhibitory effect of dipyridamole (Dipy) on SPA was studied: (a) 10 microM at each time interval; (b) 0.5-100 microM at 3 and 30 minutes and (c) 15 microM in combination with 100 microM adenosine, 8 microM 2-chloroadenosine (2Clad, an ADP receptor blocker) and 50 microM aspirin. There was a rapid decrease in the inhibitory effect of Dipy with the time after venepuncture; the correlation coefficient was -0.533. At all the concentrations studied, Dipy was more effective at 3 minutes than at 30 minutes after venepuncture. A combination of Dipy with adenosine, 2ClAd or aspirin was a more effective inhibitor of SPA than either drug alone. However, when 15 microM Dipy and 10 microM Ad were added together, the inhibitory effect of Dipy was not increased significantly, suggesting that Dipy inhibits platelet aggregation independent of Ad. The increase in SPA with the time after venepuncture was abolished when blood was taken directly into the anticoagulant containing 5 microM 2ClAd. It is suggested that ADP released from the red blood cells is responsible for the increased platelet aggregability with the time after venepuncture and makes a serious contribution to the artifacts of in vitro platelet function studies.


Saikosaponin a functions as anti-epileptic effect in pentylenetetrazol induced rats through inhibiting mTOR signaling pathway.
Saikosaponin a (SSa), which is one major bioactive compound isolated from radix bupleuri, has been demonstrated to exhibit the properties of anticonvulsant and antiepileptic in few reports. This study aims to clarify the molecular mechanism by which SSa protects against pentylenetetrazol (PTZ) induced epileptic seizure. PTZ induced rat and hippocampal neuron were established. Treated rats or hippocampal neuron with SSa, and mTOR, P70S6K, IL-1β and TNF-α were then determined. In PTZ induced rat, SSa significantly reduced seizure severity and duration while markedly elevated seizure latency, and it also down-regulated hippocampal p-mTOR, p-70S6K, L-1β and TNF-α expression. In hippocampal neurons exposed to PTZ, p-mTOR and p-70S6K expression levels were also decreased by SSa. Pre-incubated hippocampal neurons with leucine, an mTOR agonist, reversed the effects of SSa on decreasing cytokines expression and inhibiting cell apoptosis. The treatment of mTOR inhibitor rapamycin prevented against the increase of cytokines expression and hippocampal neuron apoptosis induced by PTZ. Leucine also canceled the alleviation of seizures and induction of hippocampal caspase-3 activity in PTZ induced rat with the treatment of SSa. SSa protects against PTZ induced epileptic seizure and hippocampal neuron apoptosis through inhibiting mTOR signaling pathway.


The concentration of potentially toxic elements (PTEs) in drinking water from Shiraz, Iran: a health risk assessment of samples.
The existence of potentially toxic elements (PTEs) in water bodies has posed a menace to human health. Thus, water resources should be protected from PTEs, and their effect on the exposed population should be investigated. In the present investigation, the concentrations of PTEs such as lead (Pb), mercury (Hg), manganese (Mn), and iron(Fe) in the drinking water of Shiraz, Iran, were determined for the first time. In addition, hazard quotient, hazard index, cancer risk, and sensitivity analysis were applied to estimate the noncarcinogenic and carcinogenic impacts of Pb, Hg, Mn, and Fe on exposed children and adults through ingestion. The mean concentrations (µg/L) of Pb, Hg, Mn, and Fe were 0.36, 0.32, 2.28, and 8.72, respectively, in winter and 0.50, 0.20, 0.55, and 10.36, respectively, in summer. The results displayed that Fe concentration was more than the other PTEs. PTE concentrations were lower than the standard values of the Environment Protection Agency and World Health Organization. Values of the degree of contamination and heavy metal pollution index for lead, mercury, manganese, and iron were significantly low (< 1) and excellent (< 50), respectively. Based on the Spearman rank correlation analysis, positive and negative relationships were observed in the present study. The observations of the health risk assessment demonstrated that mercury, lead, iron, and manganese had an acceptable level of noncarcinogenic harmful health risk in exposed children and adults (hazard quotients < 1 and hazard index < 1). The carcinogenic risk of lead was low (< E - 06), which can be neglected. Monte Carlo simulation showed that water intake rate and mercury concentration were the most critical parameters in the hazard index for children and adults. Lead concentration was also the most crucial factor in the cancer risk analysis. The results of the present study proved that the drinking water of Shiraz is safe and healthy and can be confidently consumed by people.


Electron transfer from triplet thymine and thymidine to lipoic acid.
Electron transfer from the triplet excited state of thymine or thymidine to the disulphide compound lipoic acid (RSSR) was studied using KrF laser flash photolysis (248 nm, 20 ns). The electron transfer reaction rate constants, measured at 310 nm, were determined to be 1.3 x 10(10) M-1 s-1 and 6.9 x 10(9) M-1 s-1 for thymine and thymidine respectively. The transient absorbance at 400 nm in the presence of the quencher is attributed to the anion radical of lipoic acid.


Development of a Pediatric Behavioral Early Warning Scale (Pedi-BEWS) for Children.
Background and Purpose: The prevention of aggressive behavior in child and adolescent inpatient settings is essential. However, it can be difficult to prompt a quick appropriate intervention without an early warning scale for the prevention and management of behavioral emergencies in a psychiatric inpatient adolescent unit. Behavioral emergencies often result in restraint/seclusion and/or administration of psychotropic medications. The objectives are to develop and evaluate the Pediatric Behavioral Early Warning Scale (Pedi-BEWS) as a screening tool to prompt quick appropriate interventions and to decrease incidents of restraint/seclusion and/or administration of psychotropic medications. Methods: A two-phase methodological design was applied. A total of 447 inpatients and 21 nurses were used to test internal consistency reliability and validity. Results: Receiver operating characteristic curve areas for the developed instrument were calculated to be 0.890 (cognition), 0.959 (affect), and 0.951 (behavior). The overall Pedi-BEWS indicated high reliability (Cronbach's alpha = .98). Conclusions: The Pedi-BEWS shows high internal consistency and validity. The use of the newly developed tool may reduce or eliminate episodes of seclusion and restraint for pediatric patients with a behavioral problem. In addition, the tool has the potential to enhance psychiatric nurses' assessment skills and competencies. The use of the newly developed Pedi-BEWS can promote an appropriate and timely nursing assessment and intervention before the deterioration in psychiatric nursing practice.


Tumor suppression, dose-limiting toxicity and wellbeing with the fetal estrogen estetrol in patients with advanced breast cancer.
The aim of this study (the ABCE4 study) was to assess dose-limiting toxicity (DLT), safety, tolerability and preliminary efficacy of high doses of the fetal estrogen estetrol (E4) in postmenopausal patients with heavily pretreated, locally advanced and/or metastatic ER+/HER2-breast cancer, resistant to anti-estrogens. This was a multicenter, open-label, phase IB/IIA, dose-escalation study with a 3 + 3 cohort design, whereby successive cohorts of three patients received 20 mg, 40 mg or 60 mg E4 per day for 12 weeks by oral administration. DLTs, safety and wellbeing were evaluated after 4, 8 and 12 weeks of treatment. Anti-tumor effects were investigated by computer tomography scanning and evaluated according to RECIST criteria before and after 12 weeks of treatment. Wellbeing was judged weekly by the investigator and by quality-of-life questionnaires by the patients. In view of the small number of patients, no statistical testing was performed. All 12 patients enrolled had progressive, heavily pre-treated advanced breast cancer. No treatment-related serious adverse events or DLTs occurred during the first 4 weeks of E4 treatment allowing the investigation of all three doses. Five of nine patients completing 12 weeks of E4 treatment showed objective anti-tumor effects and six of nine patients reported improved wellbeing. High doses of estetrol seem to be safe and are well tolerated during 12 weeks of treatment without dose-limiting toxicity and with anti-tumor effects in five of nine heavily treated patients with progressive, anti-estrogen resistant, advanced breast cancer.


Modulation of guanine nucleotide effects on the insulin receptor by MgCl2.
Insulin binding to partially purified rat adipocyte insulin receptors is inhibited approximately 40-60 percent by 1 mM GTP-gamma-S in the presence of 2 mM MgCl2. However, in the presence of 10 mM MgCl2, GTP-gamma-S does not inhibit binding. Increasing MgCl2 from 0.5 to 10 mM enhances the phosphorylation of calmodulin catalyzed by the insulin receptor but also reduces the inhibition seen with 500 microM GTP-gamma-S. The reversal of the GTP-gamma-S-induced inhibition of calmodulin phosphorylation by high concentrations of MgCl2 appears to be due to an effect on the calmodulin molecule since MgCl2 has little effect on the inhibition of phosphorylation of histone Hf2b or poly (Glu4, Tyr1) induced by GTP-gamma-S. Our data suggest that there are at least two GTP-binding proteins associated with the insulin receptor, one that regulates insulin binding and is modulated by MgCl2 and one that regulates substrate phosphorylation and/or receptor-substrate coupling and is not altered by MgCl2.


Discovery of novel prostaglandin analogs as potent and selective EP2/EP4 dual agonists.
To identify potent EP2/EP4 dual agonists with excellent subtype selectivity, a series of γ-lactam prostaglandin E analogs bearing a 16-phenyl ω-chain were synthesized and evaluated. Structural hybridization of 1 and 2, followed by more detailed chemical modification of the benzoic acid moiety, led us to the discovery of a 2-mercaptothiazole-4-carboxylic acid analog 3 as the optimal compound in the series. An isomer of this compound, the 2-mercaptothiazole-5-carboxylic acid analog 13, showed 34-fold and 13-fold less potent EP2 and EP4 receptor affinities, respectively. Structure activity relationship data from an in vitro mouse receptor binding assay are presented. Continued evaluation in an in vivo rat model of another 2-mercaptothiazole-4-carboxylic acid analog 17, optimized for sustained compound release from PLGA microspheres, demonstrated its effectiveness in a rat bone fracture-healing model following topical administration.


Interleukin-1: effects on rabbit ileal mucosal ion transport in vitro.
Effects of recombinant human interleukin-1 (IL-1) on electrical properties and unidirectional fluxes of Na+ and Cl- in rabbit ileum were examined in vitro employing the Ussing chamber technique. Serosal but not mucosal addition of Il-1 increased short-circuit current (Isc) in a concentration-dependent manner with a maximal effect occurring at 5 ng/ml and a half-maximal effect at 0.9 ng/ml. The time required for a maximal effect was between 30 and 40 min. Flux measurements revealed that serosal IL-1 reduced Na+ and Cl- absorption. The increase in Isc elicited by IL-1 was not altered by pretreatment of tissues with serosal atropine, tetrodotoxin, the H1-histamine receptor antagonist, mepyramine or removal of Ca2+ from the serosal bathing solution. The effects of IL-1 were inhibited by removal of Cl- from the bathing solutions, or by indomethacin or piroxicam. Levels of immunoreactive thromboxane B2 and 6-keto-PGF1 alpha and PGE2 were not significantly altered by IL-1. These results demonstrate that IL-1 can inhibit Na+ and Cl- absorption in rabbit ileum.


Umbilical cord mesenchymal stromal cell transplantations: A systemic analysis of clinical trials.
The advances and success of umbilical cord-derived mesenchymal stromal cells (UC-MSCs) in experimental disease animal models have fueled the development of targeted therapies in humans. The therapeutic potential of allogeneic transplantation of UC-MSCs has been under examination since 2009. The purpose of this systematic analysis was to review the published results, limitations and obstacles for UC-MSC transplantation. An extensive search strategy was applied to the published literature, 93 peer-reviewed full-text articles and abstracts were found published by early August 2017 that investigated the safety, efficacy and feasibility of UC-MSCs in 2001 patients with 53 distinct pathologies including many systemic/local, acute/chronic conditions. Few data were extracted from the abstracts and/or Chinese-written articles (n = 7, 8%). Importantly, no long-term adverse effects, tumor formation or cell rejection were reported. All studies noted certain degrees of therapeutic benefit as evidenced by clinical symptoms and/or laboratory findings. Thirty-seven percent (n = 34) of studies were found published as a single case (n = 10; 11%) or 2-10 case reports (n = 24; 26%) with no control group. Due to the nature of many stem cell-based studies, the majority of patients also received conventional therapy regimens, which obscured the pure efficacy of the cells transplanted. Randomized, blind, phase 1/2 trials with control groups (placebo-controlled) showed more plausible results. Given that most UC-MSC trials are early phase, the internationally recognized cell isolation and preparation standards should be extended to future phase 2/3 trials to reach more convincing conclusions regarding the safety and efficacy of UC-MSC therapies.


Exercise preferences among emerging adults: Do men and women want different things?
The purpose of this study was to examine exercise preferences of college students (CS), and explore potential gender differences to inform interventions. Participants (N = 187, 18-25 years) completed an online survey. Descriptive statistics were conducted to characterize preferences for exercise type and mode of delivery, followed by χ2 tests to assess potential gender differences. CS preferred a combination of moderate intensity cardio and strength training. Most CS preferred exercising on their own with guidance from a program. Men preferred intense strength training at higher rates than women (p < .001). Findings can inform tailored recruitment messaging and lifestyle interventions for this high-risk population.


Enhanced electric control of magnetic anisotropy via high thermal resistance capping layers in magnetic tunnel junctions.
We studied nonlinear magnetic anisotropy changes to the DC bias voltage of magnetic tunnel junctions (MTJs) with capping layers of different thermal resistances. We found that increasing the thickness of MgO capping layers (in the range 0.3-0.5 nm) in MTJs enhances the Joule heating-induced magnetic anisotropy change, which indicates an enhancement of the interfacial thermal resistance at the FeB|MgO capping layer interface. This enhanced interfacial thermal resistance may be attributed to roughness at the FeB|MgO interface. Moreover, we observed a larger power-driven magnetic anisotropy change of 3.21 µJ W-1m-1 in the MTJ with a composite MgO (0.3 nm)|W (2 nm)|MgO (0.4 nm) capping layer. This research supports methods of efficient spin manipulation of spintronic devices such as microwave devices and magnetic memories.


Protein and lipid lateral diffusion in normal and Rous sarcoma virus transformed chick embryo fibroblasts.
We measured the lateral diffusion of the fluorescent lipid analogue dioctadecylindocarbocyanine iodide (DiI) and of membrane glycoproteins labeled with tetramethylrhodamine (TRITC) succinyl concanavalin A (SConA) via fluorescence photobleaching recovery (FPR) at selected times during a temperature downshift experiment on transformation-defective temperature-sensitive (td-ts) Rous sarcoma virus (RSV) NY68-transformed chicken embryo fibroblasts (CEF) and on identically treated CEF and RSV-transformed CEF. There were no significant differences in the lateral diffusion in DiI at any of the times measured. The lateral diffusion of TRITC-SConA on the RSV-transformed CEF, (1.32 +/- 0.12).10(-10) cm2 s-1, was approximately two times faster than that observed in normal CEF, (0.61 +/- 0.06).10(-10) cm2 s-1. In the cells undergoing RSV NY68-mediated transformation, TRITC-SConA diffusion increased over a 24-h period from a value comparable to that observed in normal CEF, (0.72 +/- 0.13).10(-10) cm2 s-1 to a value comparable to the RSV-CEF transformed cells, (1.74 +/- 0.20).10(-10) cm2 s-1. All diffusion measurements reported were made at the permissive temperature for RSV-NY68 (35 degrees C) unless stated otherwise. The changes in the lateral diffusion of TRITC-SConA occurred between the fifth and twelfth hour of the downshift course and could be associated with cytoskeletal disruption and/or fibronectin degradation, both known to occur at this time in RSV-transformed cells. To assess the contribution of extracellular matrix (ECM) degradation, SConA mobility was measured in normal and RSV-transformed cells treated with trypsin. This treatment increased SConA mobility approximately 4-fold in the normal cells relative to untreated controls and only 2-fold in the RSV-CEF transformed cells. No significant difference in SConA mobility between trypsinized spherical normal and transformed cells was apparent.


A new vitamin D analog, 2MD, restores trabecular and cortical bone mass and strength in ovariectomized rats with established osteopenia.
An orally active, highly potent analog of 1alpha,25-dihydroxyvitamin D3, 2MD, restores trabecular and cortical bone mass and strength by stimulating periosteal bone formation and decreasing trabecular bone resorption in OVX rats with established osteopenia. The purposes of this study were to determine the effects of long-term treatment with 2-methylene-19-nor-(20S)-1alpha,25(OH)2D3 (2MD) on restoring bone mass and bone strength in ovariectomized (OVX) rats with established osteopenia and 2MD effects on bone formation and bone resorption on trabecular and cortical bone surfaces. Sprague-Dawley female rats were sham-operated (sham) or OVX at 4 months of age. Beginning at 8 weeks after OVX, OVX rats were orally dosed with 2MD at 0.5, 1, 2.5, 5, or 10 ng/kg/day for 16 weeks. Serum calcium was measured at 6, 13, and 16 weeks after treatment, and bone mass and structure, bone formation, bone resorption, and bone strength were determined at the end of the study. Serum calcium did not change significantly with 2MD at 0.5 or 1 ng/kg/day, whereas it significantly increased at 2.5, 5, or 10 ng/kg/day. 2MD significantly and dose-dependently increased total body BMD, total BMC, and stiffness of femoral shaft (FS), maximal load and stiffness of femoral neck, and toughness of the fifth lumbar vertebral body (L5) at all doses compared with OVX controls. In 2MD-treated OVX rats, there was a dose-dependent increase in total BMD and total BMC of the distal femoral metaphysis (DFM), trabecular bone volume of L3, ultimate strength and stiffness of L5, and maximal load of FS compared with OVX controls at dosages>or=1 ng/kg/day. At dosages>2.5 ng/kg/day, most of the bone mass and bone strength related parameters were significantly higher in 2MD-treated OVX rats compared with sham controls. Bone histomorphometric analysis of L3 showed dose-dependent decreases in osteoclast number and osteoclast surface on trabecular bone surface and a dose-dependent increase in periosteal bone formation associated with 2MD treatment. 2MD not only restored both trabecular and cortical bone mass but also added bone to the osteopenic OVX rats beyond that of sham controls by stimulating bone formation on the periosteal surface and decreasing bone resorption on the trabecular surface. 2MD increased bone mass and strength at doses that did not induced hypercalcemia.


Animal model for cutaneous leishmaniasis.
Using cutaneous leishmaniasis of mice, the existence of so-called T helper (Th) cells type 1 and type 2 had been identified more than 20 years ago. Nowadays, it is well accepted that additional T cell populations as well as B cell-mediated immunity is required for immunity against Leishmania major. Finally, using inbred mouse strains, the relevance of genetical factors that influence anti-pathogen immunity as well as elements of the skin-immune system have been identified. This protocol describes a model for murine experimental leishmaniasis that tries to mimic natural parasite transmission by several means: (1) utilization of only infectious-stage parasites that are found in sand fly saliva, (2) intradermal inoculation, and (3) infection with only 1,000 parasites similar to the numbers inoculated by an infected sand fly.


Effects of 17-AAG on the cell cycle and apoptosis of H446 cells and the associated mechanisms.
As a heat shock protein 90 inhibitor, 17-allylamino-17‑demethoxygeldanamycin (17-AAG) has been studied in numerous types of cancer, however the effects of 17-AAG on apoptosis and the cell cycle of H446 cells remain unclear. In the current study, the MTT method was used to evaluate the inhibitory effects of different durations and doses of 17‑AAG treatment on the proliferation of H446 cells. The cells were stained with Annexin-fluorescein isothiocyanate/propidium iodide and measured by flow cytometry, and the gene and protein expression levels of signal transducer and activator of transcription 3 (STAT3), survivin, cyclin D1, cyt‑C, caspase 9 and caspase 3 were determined by reverse transcription‑quantitative polymerase chain reaction and western blot analysis. The results indicated that with treatment with 1.25‑20 mg/l 17‑AAG for 24 and 48 h, significant inhibition of H446 cell proliferation was observed in a time‑ and dose‑dependent manner. With treatment of 3.125, 6.25 and 12.5 mg/l 17‑AAG for 48 h, significant apoptosis and cell cycle arrest was observed. The results indicated that the gene and protein expression levels of STAT3, survivin and cyclin D1 were downregulated, and cyt‑C, caspase 9 and caspase 3 were upregulated by 17‑AAG in a dose-dependent manner when the cells were treated with 3.125 and 6.25 mg/l 17-AAG for 48 h. The results indicated that 17‑AAG is able to inhibit the cell proliferation, induce apoptosis and G2/M arrest and downregulate the gene and protein expression levels of STAT3, survivin and cyclin D1, and upregulate gene and protein expression of cyt‑C, caspase 9, caspase 3.


HIV-1 infection of neurons might account for progressive HIV-1-associated encephalopathy in children.
Direct and productive infection of neurons in vivo is still a matter of debate, although in vitro experiments have demonstrated that immature neuronal cells can be productively infected by various human immunodeficiency virus (HIV) strains. To address this controversy we have analyzed, using light microscopy and in situ hybridization (ISH), HIV-1 infected cells in brain tissue from four pediatric cases of HIV-1-associated encephalopathy (EP). HIV-1 RNA-expressing cells--therefore, actively infected cells--were detected by ISH in different amounts in all brain specimens from the four children. They mainly correspond to glial cells. However, in two of the four children, who had severe progressive EP, but not in the other two, who had the static form, HIV-1-infected neurons were clearly observed in the cortical brain samples. These results provide initial evidence that HIV-1 can actively infect neurons in vivo in children and show a cortical involvement of HIV brain infection in clear correlation with the clinical EP symptoms.


Safety of Hydrocortisone Premedication Discontinuation in Patients with Inflammatory Bowel Disease on Maintenance Therapy with Infliximab: a Prospective Clinical and Pharmacological Study.
Hydrocortisone premedication reduces the risk of antibodies to infliximab [ATIs] formation in patients receiving infliximab [IFX] therapy for inflammatory bowel disease [IBD]. We aimed to determine the safety of hydrocortisone premedication withdrawal in IBD patients with sustained clinical response on maintenance therapy with IFX. We performed an observational prospective pharmacoclinical study in a tertiary referral centre, including all consecutive IBD outpatients with no previous IFX infusion reaction and in clinical remission on maintenance IFX [alone or in combination therapy] for at least 6 months. This cohort was followed for 1 year after discontinuation of hydrocortisone premedication. Among the 268 IBD outpatients, 95 patients met the inclusion criteria [mean age 38 years; 64% male; 80% Crohn's disease; 45% combination therapy]. The median IFX duration was 5 years [0.54-14] with a mean infused dose of 533 mg [200-1000] and a mean interval duration of 7.9 weeks [4-10]. None of the patients developed permanent ATIs or infusion-related reaction at 1 year. Four patients developed transient ATIs without loss of clinical response. There was no significant variation of infliximab serum trough levels [5.5 µg/mL vs 5.9 µg/mL] measured at the time of the three IFX infusions before and after hydrocortisone withdrawal. Loss of response rate to IFX was 18% at 1 year. Hydrocortisone discontinuation is safe in IBD patients with sustained clinical remission on maintenance therapy with IFX. Our data suggest that routine premedication with hydrocortisone is unnecessary in patients in prolonged remission under IFX maintenance therapy.


Depression roundtable: Is there a role for BoNT?
Depression can occur in the context of major depressive disorder or bipolar disorder. There are many effective and well-tolerated treatment options for most patients experiencing major depressive episodes, but for patients with treatment-resistant major depressive disorder or bipolar depression, current pharmacologic and non-pharmacologic options can be less efficacious, well tolerated, or accessible. Botulinum neurotoxin (BoNT) offers a novel approach to treating depression that is both safe and well-tolerated. Several potential mechanisms of action in depression are theorized, and studies support the efficacy of BoNT in major depressive disorder. Early data suggests that BoNT may be efficacious in bipolar depression and further study is warranted.


Interpersonal distance modulates the influence of social observation on prosocial behaviour: An event-related potential (ERP) study.
This study aimed to explore the influence of interpersonal distance and social observation on prosocial behaviour. Participants were instructed to make costly prosocial decisions towards different interpersonal distance targets (friends, acquaintances or strangers) under the (non) observable condition, with simultaneous electroencephalogram recording. The behavioural results demonstrated that participants made more prosocial choices-unaffected by social observation-towards friends than towards acquaintances and strangers; nonetheless, participants made more prosocial choices towards acquaintances and strangers under the observable than under the non-observable condition. Event-related potential results showed that when participants made prosocial decisions towards friends, the P3 and N2 amplitudes remained unchanged between the observable and non-observable conditions; however, when participants made prosocial decisions towards acquaintances and strangers, a smaller N2 and a larger P3 were observed under the observable than those under the non-observable condition. These findings suggest that prosocial behaviour towards friends is driven by social preferences regarding the welfare of others, regardless of the possibility of reputation management. However, prosocial behaviour towards acquaintances and strangers might be motivated by a positive reputation, because individuals may bear in mind the potential future benefits.


Microwave dielectric characterisation of 3D-printed BaTiO3/ABS polymer composites.
3D printing is used extensively in product prototyping and continues to emerge as a viable option for the direct manufacture of final parts. It is known that dielectric materials with relatively high real permittivity-which are required in important technology sectors such as electronics and communications-may be 3D printed using a variety of techniques. Among these, the fused deposition of polymer composites is particularly straightforward but the range of dielectric permittivities available through commercial feedstock materials is limited. Here we report on the fabrication of a series of composites composed of various loadings of BaTiO3 microparticles in the polymer acrylonitrile butadiene styrene (ABS), which may be used with a commercial desktop 3D printer to produce printed parts containing user-defined regions with high permittivity. The microwave dielectric properties of printed parts with BaTiO3 loadings up to 70 wt% were characterised using a 15 GHz split post dielectric resonator and had real relative permittivities in the range 2.6-8.7 and loss tangents in the range 0.005-0.027. Permittivities were reproducible over the entire process, and matched those of bulk unprinted materials, to within ~1%, suggesting that the technique may be employed as a viable manufacturing process for dielectric composites.


Association of resistin with impaired membrane fluidity of red blood cells in hypertensive and normotensive men: an electron paramagnetic resonance study.
Abnormalities in physical properties of the cell membranes may strongly be linked to hypertension. Recent evidence indicates that resistin may actively participate in the pathophysiology of insulin resistance, diabetes mellitus, hypertension and other circulatory disorders. The present study was undertaken to investigate the possible relationships among plasma resistin, oxidative stress and membrane fluidity (a reciprocal value of membrane microviscosity) in hypertension. We measured the membrane fluidity of red blood cells (RBCs) in hypertensive and normotensive men using an electron paramagnetic resonance (EPR) and spin-labeling method. The order parameter (S) for the spin-label agents (5-nitroxide stearate) in EPR spectra of red blood cell (RBC) membranes was significantly higher in hypertensive men than in normotensive men, indicating that membrane fluidity was decreased in hypertension. Plasma resistin levels were correlated with systolic blood pressure and 8-iso-prostaglandin F2α levels (an index of oxidative stress). Furthermore, the order parameter (S) of RBCs significantly correlated with plasma resistin and plasma 8-isoPG F2α, suggesting that reduced membrane fluidity of RBCs might be associated with hyperresistinemia and increased oxidative stress. Multivariate regression analysis showed that, after adjustment for confounding factors, plasma resistin might be an independent determinant of membrane fluidity of RBCs. The EPR study suggests that resistin might have a close correlation with impaired rheologic behavior of RBCs and microcirculatory dysfunction in hypertension, at least in part, via an oxidative stress-dependent mechanism.


Purification of total RNA from human stool samples.
While colonoscopy may detect early-stage colon tumors, a less invasive and more cost-effective technique would be beneficial. Stool, which picks up sloughed-off colonic epithelial cells, would be ideal for sampling the mucosa; shed tumor cells may display alterations in gene expression observed in intact tumors. It is first necessary, however, to show that RNA can be isolated from human feces and that this RNA contains human gene transcripts. We have therefore developed a method for the isolation of total RNA from freshly passed human stool, consisting of lysis in chaotropic agents, repeated extraction with phenol and phenol-chloroform, and absorption with an RNA-binding resin. After treatment with RNase-free DNase I, we assayed these preparations for the presence of human RNA by quantitative slot blotting, northern blotting, and reverse transcription-polymerase chain reaction (RT-PCR). We obtained 5-30 microg RNA per gram of stool from cancer patients, and about 5 microg RNA per gram of control stool. Quantitative slot blotting showed that about 10% of this RNA was of human origin. Both northern blotting and RT-PCR demonstrated the presence of human RNA in these samples. To unambiguously demonstrate the isolation of RNA from stool, we incubated a mixture of rat cells and control human stool at 37 degrees C for up to 24 hr. RT-PCR of the RNA isolated from this sample clearly revealed the presence of rat-specific mRNA. These experiments indicate that RNA can be isolated from human stool and that message encoded by human genes can be assayed in these preparations. This procedure may provide a powerful tool to identify patients at risk for colon cancer.


[Elderly patients and atrial fibrillation: age alone should not be a contra-indication for anticoagulant treatment].
A significant proportion of elderly patients are not properly treated with anticoagulants. Here we describe the pros and cons of anticoagulant treatment in elderly patients with atrial fibrillation, illustrated by two cases. We discuss the sometimes difficult treatment decisions to be made in frail elderly patients, especially after they have suffered bleeding complications. Aspirin is now considered obsolete in the treatment of atrial fibrillation in the elderly.


Merkel Cell Carcinoma Therapeutic Update.
Merkel cell carcinoma (MCC) is a rare and aggressive neuroendocrine tumor of the skin. Early-stage disease can be cured with surgical resection and radiotherapy (RT). Sentinel lymph node biopsy (SLNB) is an important staging tool, as a microscopic MCC is frequently identified. Adjuvant RT to the primary excision site and regional lymph node bed may improve locoregional control. However, newer studies confirm that patients with biopsy-negative sentinel lymph nodes may not benefit from regional RT. Advanced MCC currently lacks a highly effective treatment as responses to chemotherapy are not durable. Recent work suggests that immunotherapy targeting the programmed cell death receptor 1/programmed cell death ligand 1 (PD-1/PD-L1) checkpoint holds great promise in treating advanced MCC and may provide durable responses in a portion of patients. At the same time, high-throughput sequencing studies have demonstrated significant differences in the mutational profiles of tumors with and without the Merkel cell polyomavirus (MCV). An important secondary endpoint in the ongoing immunotherapy trials for MCC will be determining if there is a response difference between the virus-positive MCC tumors that typically lack a large mutational burden and the virus-negative tumors that have a large number of somatic mutations and predicted tumor neoantigens. Interestingly, sequencing studies have failed to identify a highly recurrent activated driver pathway in the majority of MCC tumors. This may explain why targeted therapies can demonstrate exceptional responses in case reports but fail when treating all comers with MCC. Ultimately, a precision medicine approach may be more appropriate for treating MCC, where identified driver mutations are used to direct targeted therapies. At a minimum, stratifying patients in future clinical trials based on tumor viral status should be considered as virus-negative tumors are more likely to harbor activating driver mutations.


Children, adolescents and trauma.
This paper presents an overview of the psychological effects of trauma on children and adolescents with specific attention to the epidemiology of traumatic experiences, risk factors, developmental effects, the types of traumatic experiences albeit a single event trauma or a chronic process trauma, the spectrum of clinical presentations, psychiatric and psychological comorbidities as well as assessment and therapeutic principles. In addition to the child's psychological response to the traumatic stressor communities are often devastated by natural or man made disasters. The interactions between the child's response with the family and community response as well as community interventions are discussed.


Immunoinformatics Analysis of Citrullinated Antigen as Potential Multi-peptide Lung Cancer Vaccine Candidates for Indonesian Population.
Non-small-cell lung cancer (NSCLC) is the most common lung cancer which has the highest mortality rate in Indonesia. One of the trends in treating cancer is by utilizing peptide vaccines, an immunotherapeutic approach that aims to stimulate the cell-mediated adaptive immune system to recognize cancer-associated peptides. Currently, no peptide vaccines are available in the market for NSCLC treatment. Therefore, this project aims to develop a multi-epitope peptide-based vaccine for NSCLC utilizing citrullinated peptides. Citrullination is a post-translational modification that occurs in cancer cells during autophagy that functions to induce immune responses towards modified self-epitopes such as tumor cells, through activation of PAD enzymes within the APC and target cells. It was found that introducing a common citrullinated neo-antigen peptide such as vimentin and enolase to the immune system could stimulate a higher specific CD4+ T cell response against NSCLC. Moreover, carcinoembryonic antigen (CEA), an antigen that is highly expressed in cancer cells, is also added to increase the vaccine's specificity and to mobilize both CD4+ and CD8+ T cells. These antigens bind strongly to the MHC Class II alleles such as HLA-DRB1*07:01 and HLA-DRB*11:01, which are predominant alleles in Indonesian populations. Through in silico approach, the peptides generated from CEA, citrullinated vimentin and enolase, were analyzed for their MHC binding strength, immunogenicity, ability to induce IFNγ response, and population coverage. It is expected that the immunodominant antigens presentation is able to induce a potent immune response in NSCLC patients in Indonesia, resulting in tumor eradication.


Mammalian oocytes exhibit specific recognition of the RGD (Arg-Gly-Asp) tripeptide and express oolemmal integrins.
Integrins are a family of cell adhesion receptors involved in many cell-cell and cell-matrix interactions. Some of these heterodimeric receptors, such as alpha 5 beta 1 and alpha v beta 1, specifically recognize the amino acid sequence Arg-Gly-Asp (RGD) within their ligands. The RGD sequence is found in fibronectin, vitronectin, and other extracellular matrix proteins. Our results demonstrate that the oolemmas of eggs from human and several other mammalian species contain receptors capable of binding to RGD ligands, and that integrin subunits are expressed by oocytes. Four distinct techniques were utilized to identify the presence of functional integrins on mammalian eggs. RGD-binding receptors were detected on the surfaces of zona-free eggs from all species tested. Covaspheres coated with PepTite-2000, which contains RGD, bound to the eggs and formed rosettes. Rosetting was competitively inhibited by PepTite-2000 and by GRGDTP, a soluble RGD peptide, but not by RGES. An ELISA using polyclonal antibodies directed against the cytoplasmic tails of the integrin subunits identified the integrin subunits alpha 5, beta 1, and alpha v, but not beta 3, in detergent extracts of Syrian hamster eggs. A dot blot confirmed the presence of alpha v in hamster egg lysates. Finally, the integrin subunits alpha 2, alpha 5, alpha 6, but not alpha 4, were detected on the surfaces of zona-free eggs from human and Syrian hamster. Immunobeads coated with monoclonal antibodies specific for alpha 2, alpha 5, and alpha 6 bound to the eggs and formed rosettes.(ABSTRACT TRUNCATED AT 250 WORDS)


Polymorphisms in the gene encoding adiponectin receptor 1 are associated with insulin resistance and high liver fat.
The adipokine adiponectin has insulin-sensitising, anti-atherogenic and anti-inflammatory properties. Recently, the genes for mouse and human adiponectin receptor-1 (ADIPOR1) and -2 (ADIPOR2) have been cloned. The aim of this study was to investigate whether genetic variants of the genes encoding ADIPOR1 and ADIPOR2 play a role in human metabolism. We screened ADIPOR1 and ADIPOR2 for polymorphisms and determined their association with glucose metabolism, lipid metabolism, an atherogenic lipid profile and inflammatory markers in 502 non-diabetic subjects. A subgroup participated in a longitudinal study; these subjects received diet counselling and increased their physical activity. We identified six variants of ADIPOR1 and seven variants of ADIPOR2. A single-nucleotide polymorphism (SNP) in the putative promoter region 8503 bp upstream of the translational start codon (-8503 G/A) of ADIPOR1 (frequency of allele A=0.31) was in almost complete linkage disequilibrium with another SNP (-1927 T/C) in intron 1. Subjects carrying the -8503 A and -1927 C alleles had lower insulin sensitivity, as estimated from a 75 g OGTT (p=0.04) and determined during a euglycaemic clamp (n=295, p=0.04); they also had higher HbA(1)c levels (p=0.02) and, although the difference was not statistically significant, higher liver fat (n=85, determined by proton magnetic resonance spectroscopy, p=0.056) (all p values are adjusted for age, sex and percentage of body fat). In the longitudinal study (n=45), the -8503 A and -1927 C alleles were associated with lower insulin sensitivity (p=0.03) and higher liver fat (p=0.02) at follow-up compared with the -8503 G and -1927 T alleles, independently of basal measurements, sex and baseline and follow-up percentage of body fat. The present findings suggest that the -8503 G/A SNP in the promoter or the -1927 T/C SNP in intron 1 of ADIPOR1 may affect insulin sensitivity and liver fat in humans.


Pleasure and suffering of nursing in COVID-19 hospital units: between disenchantment and formation of meanings.
to analyze the experiences of pleasure and suffering of nursing workers in COVID-19 hospital units. a multicenter, qualitative study, developed with 35 nursing workers from COVID-19 units in seven hospitals in southern Brazil. Data were produced through semi-structured interviews, submitted to thematic content analysis with the help of NVivo. experiences of pleasure were linked to gratification, identification with work content, positive results in care, recognition, integration with the team and personal overcoming. Suffering was revealed in daily life of deaths and losses, feelings of helplessness, team conflicts, institutional demands, professional devaluation. Workers reported disenchantment, but also strengthening the meaning of their work, highlighting frontline impacts on their mental health. in the dynamics between pleasure and suffering in nursing work in COVID-19 hospital units, elements point to the risk of psychological illness.


Surface expression and function of Cav3.2 T-type calcium channels are controlled by asparagine-linked glycosylation.
Low-voltage-activated T-type calcium channels play important roles in neuronal physiology where they control cellular excitability and synaptic transmission. Alteration in T-type channel expression has been linked to various pathophysiological conditions such as pain arising from diabetic neuropathy. In the present study, we looked at the role of asparagine (N)-linked glycosylation on human Cav3.2 T-type channel expression and function. Manipulation of N-glycans on cells expressing a recombinant Cav3.2 channel revealed that N-linked glycosylation is critical for proper functional expression of the channel. Using site-directed mutagenesis to disrupt the canonical N-linked glycosylation sites of Cav3.2 channel, we show that glycosylation at asparagine N192 is critical for channel expression at the surface, whereas glycosylation at asparagine N1466 controls channel activity. Moreover, we demonstrate that N-linked glycosylation of Cav3.2 not only controls surface expression and activity of the channel but also underlies glucose-dependent potentiation of T-type Ca(2+) current. Our data suggest that N-linked glycosylation of T-type channels may play an important role in aberrant upregulation of T-type channel activity in response to glucose elevations.


The First National Survey of Indigenous People's Health and Nutrition in Brazil: rationale, methodology, and overview of results.
Although case studies indicate that indigenous peoples in Brazil often suffer from higher morbidity and mortality rates than the national population, they were not included systematically in any previous national health survey. Reported here for the first time, the First National Survey of Indigenous People's Health and Nutrition in Brazil was conducted in 2008-2009 to obtain baseline information based on a nationwide representative sample. This paper presents the study's rationale, design and methods, and selected results. The survey sought to characterize nutritional status and other health measures in indigenous children less than 5 years of age and indigenous women from 14 to 49 years of age on the basis of a survey employing a representative probabilistic sample of the indigenous population residing in villages in Brazil, according to four major regions (North, Northeast, Central-West, and South/Southeast). Interviews, clinical measurements, and secondary data collection in the field addressed the major topics: nutritional status, prevalence of hypertension and diabetes mellitus in women, child hospitalization, prevalence of tuberculosis and malaria in women, access to health services and programs, and characteristics of the domestic economy and diet. The study obtained data for 113 villages (91.9% of the planned sample), 5,305 households (93.5%), 6,692 women (101.3%), and 6,128 children (93.1%). Multiple household variables followed a pattern of greater economic autonomy and lower socioeconomic status in the North as compared to other regions. For non-pregnant women, elevated prevalence rates were encountered for overweight (30.3%), obesity (15.8%), anemia (32.7%), and hypertension (13.2%). Among children, elevated prevalence rates were observed for height-for-age deficit (25.7%), anemia (51.2%), hospitalizations during the prior 12 months (19.3%), and diarrhea during the prior week (23.6%). The clinical-epidemiological parameters evaluated for indigenous women point to the accentuated occurrence of nutrition transition in all regions of Brazil. Many outcomes also reflected a pattern whereby indigenous women's and children's health indicators were worse than those documented for the national Brazilian population, with important regional variations. Observed disparities in health indicators underscore that basic healthcare and sanitation services are not yet as widely available in Brazil's indigenous communities as they are in the rest of the country.


Contribution of adrenal norepinephrine output to increase aortic norepinephrine during carotid sinus reflex activation in anesthetized dogs.
Changes in circulating plasma catecholamine (CA: E, epinephrine; NE, norepinephrine; and DA, dopamine) concentrations in aortic (AO) blood were investigated in relation to variable rates of CA secretion from both adrenal (ADR) glands in response to bilateral carotid artery occlusion (BLCO) in vagotomized dogs anesthetized with sodium pentobarbital. During BLCO (3 min), AO systolic pressure (AP) increased along with significant increases in ADR-CA output, renal venous (RV) CA output, as well as in AO-E and NE concentrations. A ratio of NE:E in ADR venous and AO blood did not exceed 0.42 +/- 0.09 and 1.09 +/- 0.24 upon BLCO, respectively. In contrast, the NE:E ratio in RV blood increased significantly from 5.39 +/- 0.91 to 9.78 +/- 1.31. Following adrenalectomy (ADRX), the increase in AO-NE in response to BLCO was significantly attenuated by approximately 56%, but the increase in RV-NE output was not affected by ADRX. The results show that in vagotomized dogs, NE is co-released with E from the adrenal glands upon BLCO. The data also indicate that the increase in AO-NE concentration was dependent to a similar extent on the simultaneous increases in ADR-NE output and neuronal NE release. We conclude that under conditions where the sympathoadrenal system is activated, circulating plasma NE concentration may be significantly affected by an increase in ADR-NE output. Sympathetic neuronal contributions would, thereby, be overestimated in assessing overall sympathetic nerve activity by measuring circulating NE. NE concentrations in local venous effluent from individual organs may be more reliable estimates of the sympathetic nerve activity.


Sleep EEG oscillations in neurodevelopmental disorders without intellectual disabilities.
Neurodevelopmental disorders (NDDs) are often characterised by sleep problems, and recent evidence indicates alterations of the sleep electroencephalographic (EEG) oscillations in these patients. Sleep microstructure plays a crucial role in cognitive functioning and brain maturation. In this view, modifications in sleep EEG oscillations in NDDs could further impair the cognitive maturation process in these patients. We provide an overview of sleep microstructure alterations observed in three NDDs without intellectual disabilities (attention-deficit/hyperactivity disorder, high-functioning autism/Asperger syndrome and developmental dyslexia) and their relationships with the disorders' phenomenology. For each NDD, we discuss empirical evidence of altered EEG oscillations, and we consider their interaction with patients' cognitive and behavioural functioning, with the aim to elucidate their functional meaning. We highlight the limits of the present literature and propose possible future directions while underlining the clinical relevance of the research in this field. Beyond confirming the importance of sleep management in atypically developing children, the review findings suggest that sleep EEG oscillations in NDDs could become a target for specific clinical intervention.


Retrieval of Displaced Third Molar Into the Sublingual Space Using 3-Dimensional Navigation Assistance.
Surgical removal of third molars is a common procedure undertaken by both general dentists and oral and maxillofacial surgeons. Although rare, iatrogenic displacement of the tooth or root fragments into adjacent fascial spaces is a known complication. This case report details the retrieval of displaced root tips from a mandibular third molar into the sublingual space using intraoperative navigation to minimize surgical exploration. Oral-maxillofacial surgeons should consider using 3-dimensional navigation during scheduled retrieval of displaced teeth, fragments, or foreign objects to minimize the risk of surgical complications.


Aspirin use in central retinal arterial occlusion to prevent ischaemic stroke: a retrospective cohort study in Taiwan.
To understand the efficacy of aspirin use for preventing ischaemic stroke after central retinal artery occlusion (CRAO). The retrospective cohort study was conducted using the National Health Insurance Research Database from 1998 to 2013. A population-based study. A total of 9437 participants with newly diagnosed CRAO were identified. Participants who had a previous stroke and/or retinal vascular occlusion, were aged <20 years and used aspirin 3 months before the event were excluded. There were 3778 eligible participants matched by propensity score, and they were divided into aspirin (n=434) and aspirin-naive (n=1736) groups after the matching. Cox proportional hazard models and cumulative survival curves were used to assess ischaemic stroke in the study groups, along with log-rank tests to compare group differences. Incidence of ischaemic stroke in the aspirin and aspirin-naive groups 1 year after CRAO. Of the 3778 patients with newly diagnosed CRAO, 151 (4%) had a subsequent ischaemic stroke within 1 year. The risk was especially high during the first week of the CRAO. No difference between the aspirin and aspirin-naive groups was found in risk of ischaemic stroke, haemorrhagic stroke, gastrointestinal bleeding, major bleeding, acute coronary syndrome, retinal vein occlusion, new-onset glaucoma, undergoing panretinal photocoagulation or all-cause mortality. Risk factors for ischaemic stroke within 1 year of CRAO included male gender (p=0.031; HR=1.46) and age (p=0.032; HR=1.14). Aspirin use after a CRAO showed no benefit on attenuating the risk of ischaemic stroke. The risk of ischaemic stroke was increased after CRAO especially during the first week. Male gender and age were risk factors for ischaemic stroke after CRAO.


A population-based study of health service deficits for US adults with asthma.
Asthma prevalence in the United States is higher than it is in many other countries and its impact in terms of healthcare expenditures and morbidity and mortality is staggering. In the United States, many groups bear a disproportionate burden of asthma. Understanding the epidemiology of adult asthma and deficits in health care can identify opportunities for improving care and effectively managing resources. The computed dependent variable, health service deficits, entails a lack of health insurance, not having a healthcare provider, deferring medical care because of cost, and having had no routine medical exam. Bivariate and multivariate analyses were performed on 2005 Behavioral Risk Factor Surveillance Survey data to examine the relationship between health service deficits experienced by adults with asthma and socioeconomic status (SES), race and ethnicity, and geographic locale. The variable was also calculated for each US state. Hispanic (OR = 1.594, 95% CI = 1.588-1.599) and Other/Multiracial (OR = 1.447, 95% CI = 1.441-1.452) adults with current asthma had greater odds of having a health service deficit. Rural adults with current asthma had greater odds of having a health service deficit (OR = 1.086, 95% CI = 1.083-1.089) when compared with non-rural adult residents. Low-SES (OR = 1.976, 95% CI = 1.971-1.982) and middle-SES (OR = 1.596, 95% CI = 1.592-1.600) adults with current asthma had greater odds of having a health service deficit. The percentage of current asthma adults experiencing at least one health service deficit by state ranged from a low of 28.5% (Delaware) to a high of 58.8% (Wyoming). There are clear patterns of disparity associated with health services and asthma that can help target interventions.


Natural Selection, Intracellular Bottlenecks of Virus Populations, and Viral Superinfection Exclusion.
Natural selection acts on cellular organisms by ensuring the genes responsible for an advantageous phenotype consistently reap the phenotypic advantage. This is possible because reproductive cells of these organisms are almost always haploid, separating the beneficial gene from its rival allele at every generation. How natural selection acts on plus-strand RNA viruses is unclear because these viruses frequently load host cells with numerous genome copies and replicate thousands of progeny genomes in each cell. Recent studies suggest that these viruses encode the Bottleneck, Isolate, Amplify, Select (BIAS) mechanism that blocks all but a few viral genome copies from replication, thus creating the environment in which the bottleneck-escaping viral genome copies are isolated from each other, allowing natural selection to reward beneficial mutations and purge lethal errors. This BIAS mechanism also blocks the genomes of highly homologous superinfecting viruses, thus explaining cellular-level superinfection exclusion.


Quantitative evaluation of video laryngostroboscopy: reliability of the basic parameters.
The purpose of this study was to evaluate quantitatively the basic parameters of the video laryngostroboscopy (VLS) and determine the sensitivity and specificity of these parameters discriminating healthy and pathological voice classes. Digital VLS recordings were performed for 159 individuals: 26 healthy and 133 patients. VLS variables (glottal closure, regularity, mucosal wave on the affected/healthy side, symmetry of vibration, and symmetry of image) were rated two times with the time interval of 1 year by three laryngologists. To evaluate interrater and test-retest reliability, intraclass correlation coefficients (ICCs) were calculated. To evaluate sensitivity and specificity of the VLS parameters, discriminant analysis was used. Moderate to almost perfect levels (ICC 0.46-0.90) of interrater reliability were revealed for most of the basic VLS parameters. The ICC of the interrater reliability was highest for symmetry of glottal image; the most problematic VLS parameter for rating was mucosal wave on the healthy side. ICC of the test-retest reliability were 0.71-0.95, P<0.001. An optimum system of VLS parameters discriminating normal and pathological voice subgroups with sensitivity 96.3% and specificity 100% included glottal closure and mucosal wave on the affected side. The quantitative evaluation of the VLS using basic parameters showed to be reliable in clinical settings and demonstrated high sensitivity and specificity distinguishing healthy and pathological voice patient groups.


A brief overview of the past, present, and future of the Department of General Surgery at Pietersburg and Mankweng hospitals in Limpopo Province, South Africa.
A brief overview of the Department of Surgery in the past, present and future.


Effects of replacing medium sodium by choline, caesium, or rubidium, on water and ion contents of renal cortical slices.
1. Renal cortical slices from rat, rabbit, and guinea-pig were incubated in media in which choline, caesium or rubidium replaced sodium.2. Slices of rabbit and guinea-pig renal cortex incubated in oxygenated choline Ringer decreased in volume initially and did not swell over 3 hr at 25 degrees C. There was a steady loss of potassium. Inhibition of metabolism (N(2) + 1 mM iodoacetamide) caused some swelling. Ouabain, 10 mM, in choline Ringer affected neither loss of potassium nor tissue water content.3. Slices of rat renal cortex similarly incubated in choline Ringer swelled over 3 hr at 25 degrees C whether or not metabolism was inhibited; ouabain (15 mM) affected neither tissue potassium loss nor tissue water content.4. Incubation in choline Ringer containing either 0.2 mMp-chloromercuribenzoic acid, or 1 mM ethacrynic acid increased the tissue water content of guinea-pig renal cortical slices.5. Depletion of cellular potassium (by preliminary incubation in oxygenated potassium-free sodium Ringer with 10 mM ouabain at 30 degrees C) resulted in increased tissue water content when rabbit renal cortical slices were subsequently incubated in oxygenated choline Ringer at 25 degrees C for 3 hr.6. There was no evidence of energy-dependent extrusion of water or ions from either equilibrated rat or rabbit renal cortical slices leached at 0.5 degrees C and then reincubated at 25 degrees C in choline Ringer.7. Rat and guinea-pig renal cortical slices leached at 0.5 degrees C and reincubated at 25 degrees C swelled in rubidium Ringer and in caesium Ringer. There was no evidence of energy-dependent water or ion extrusion when metabolism was restored after leaching in either of these media. Metabolizing rat slices but not guinea-pig slices swelled faster than slices whose metabolism was inhibited.8. These results lend no support to the mechano-chemical hypothesis which ascribes cellular volume regulation to a contractile mechanism squeezing isotonic extracellular fluid from the cells. Instead it is suggested that cellular water content in these experiments reflects the balance between the rate of loss of potassium (and chloride) from the cells and the rate of uptake of extracellular cation (and chloride) into the cells - these rates reflecting both the electrochemical potential gradients of the ions and membrane permeability to them. The implications in relation to the hypothesis of ouabain-insensitive cellular volume regulation are discussed.


Effects of diabetic pharmacotherapy on prolactin hormone in patients with type 2 diabetes mellitus: Bane or Boon.
Prolactin is a polypeptide hormone secreted from the anterior part of the pituitary gland which was implicated as a diabetogenic factor in the pathogenesis of type 2 diabetes mellitus (T2DM). Therefore, the aim of the present study was to evaluate prolactin serum levels in patients with T2DM regarding the effect of diabetic pharmacotherapy. Eighty patients with T2DM compared with 25 male healthy controls were recruited and divided into four groups: Group I - 29 patients with T2DM treated with metformin, Group II - 30 patients with T2DM treated with glyburide, Group III - 21 patients with T2DM treated with glyburide plus metformin, and Group IV - 25 control male healthy patients. Prolactin serum levels were high in patients with T2DM compared with controls (P < 0.01). Prolactin serum levels were higher in glyburide-treated patients compared with metformin-treated patients (P < 0.01). This study concludes that high prolactin levels in patients with T2DM are linked with diabetic complications and regarded as a beneficial phenomena to overcome IR and diabetic complications. Metformin but not glyburide reduced prolactin levels due to the improvement of insulin resistance.


A single amino-acid substitution in the iron-sulphur protein subunit of succinate dehydrogenase determines resistance to carboxin in Mycosphaerella graminicola.
A gene encoding the iron-sulphur protein (Ip) subunit of succinate dehydrogenase (Sdh, EC 126.96.36.199) from Mycosphaerella graminicola (Septoria tritici) has been cloned andsequenced. The deduced amino-acid sequence exhibited a high degree of homology to Ip subunits of Sdh from other organisms; three cysteine-rich clusters associated with the iron-sulphur centres involved in electron transport were particularly conserved. Expression studies using a synthetic green fluorescent protein (SGFP) expression vector demonstrated that the cloned DNA also contained a functional promoter region and confirmed that the deduced initiation codon could act as a translational start site. Mutants resistant to the fungicide carboxin (Cbx), a known inhibitor of Sdh, were found to contain a single amino-acid substitution in the third cysteine-rich domain of the Ip protein. These mutations resulted in the conversion of a highly conserved His residue, located in a region of the protein associated with the [3Fe-4 S] high-potential non-heme iron sulphur-redox (S3) centre, to either Tyr or Leu. AnIp gene containing the His -> Tyr mutation was constructed and shown to confer Cbx resistance following co-transformation into the Cbx-sensitive wild-type strain. This confirmed that the mutation identified by sequence analysis was responsible for determining Cbx resistance.


Great expectations: specific lexical anticipation influences the processing of spoken language.
Recently several studies have shown that people use contextual information to make predictions about the rest of the sentence or story as the text unfolds. Using event related potentials (ERPs) we tested whether these on-line predictions are based on a message-level representation of the discourse or on simple automatic activation by individual words. Subjects heard short stories that were highly constraining for one specific noun, or stories that were not specifically predictive but contained the same prime words as the predictive stories. To test whether listeners make specific predictions critical nouns were preceded by an adjective that was inflected according to, or in contrast with, the gender of the expected noun. When the message of the preceding discourse was predictive, adjectives with an unexpected gender inflection evoked a negative deflection over right-frontal electrodes between 300 and 600 ms. This effect was not present in the prime control context, indicating that the prediction mismatch does not hinge on word-based priming but is based on the actual message of the discourse. When listening to a constraining discourse people rapidly make very specific predictions about the remainder of the story, as the story unfolds. These predictions are not simply based on word-based automatic activation, but take into account the actual message of the discourse.


Interleukin-10 controls human peripheral PMN activation triggered by lipopolysaccharide.
Large amounts of anti-inflammatory mediators, such as interleukin (IL)-10, are produced and found early in the course of sepsis. We explore the role of IL-10 on neutrophil (PMN) activation/function using an in vitro model. Isolated human PMN were pre-incubated with lipopolysaccharide (LPS) and/or IL-10 for 18h. Subsequently, a second LPS exposure was performed and CD11b and CD66b up-regulation, and the reactive oxygen species (ROS) generation were measured 2h later. We found that IL-10 prevented PMN activation and the secretion of TNF-α and IL-8 induced by the first LPS contact. In the absence of IL-10, a second LPS exposure induced additive effects that were prevented by IL-10. Only ROS generation was highly affected by the blockade of PMN-secreted TNF-α or IL-8. Additionally, IL-10 prevented other possible mechanisms of LPS priming. Therefore, IL-10 modulates PMN activation preventing autocrine activating loops and priming mechanisms, rendering PMN less responsive to a second LPS exposure.


Density gradient centrifugation of cells separated from multicellular tumor spheroids.
Cells from a murine fibrosarcome (FSa) have been grown in vitro as multicell tumor spheroids (MTS). The growth rate of these MTS was determined. Following selected periods of growth, MTS were made into a single cell suspension and separated on linear density gradients of Renografin. While only 1 population of cells were separated from small spheriods (400 mum diameter), at least 3 subpopulations of tumor cells were separated and isolated from large spheroids (800 mum in diameter).


Online comprehensive two-dimensional hydrophilic interaction chromatography×reversed-phase liquid chromatography coupled with hybrid linear ion trap Orbitrap mass spectrometry for the analysis of phenolic acids in Salvia miltiorrhiza.
In the present study, a system was developed for the analysis of phenolic acids in Salvia miltiorrhiza using online comprehensive two-dimensional hydrophilic interaction chromatography and reversed-phase liquid chromatography coupled to a DAD detector and hybrid linear ion trap-Orbitrap mass spectrometry (HILIC×RP-DAD-ESI/HRMS/MSn). The system was configured based on the combination of an XBridge Amide column (150mm×4.6mm, 3.5μm) and Accucore PFP column (50mm×4.6mm, 2.6μm) for the first and second dimensions, respectively. An additional LC pump was used to dilute the eluent from the first dimension to decrease its elution strength in the second dimension. A back-flush trap column was selected as an interface to make up for the loss of efficiency and resolution due to the online dilution. Under the optimized conditions, a total of 196 peaks of polar compounds were successfully separated and detected in Salvia miltiorrhiza using the developed online HILIC×RP system, which exhibited high orthogonality (73%). The online combination of HILIC and RP provides powerful separation capability for the analysis of polar compounds in samples with complex matrices.


Correlations between osteoprotegerin serum levels and body composition parameters in patients with sleep apnea syndrome and the possible influence on cardiovascular risk.
Osteoprotegerin (OPG) is a member of the tumor necrosis factor family and a key regulator in bone turnover; it plays a role in the development of many cardiovascular diseases and may be treated as a marker of vascular damage. Bioelectrical impedance analysis (BIA) is a reliable, non-invasive and effective technique for measuring body composition. The aim of the study was to evaluate correlations between osteoprotegerin serum levels and body composition parameters in sleep apnea patients and their influence on cardiovascular risk. A total of 125 patients with newly diagnosed OSA were enrolled in the study (including 34 females). The mean age was 54.48±8.81 years, mean AHI 33.16±20.44/h and mean BMI 33.76±7.18. A control group comprised 59 healthy subjects with mean age of 51.27±12.97 years and mean BMI 29.47±5.42. All subjects underwent a nocturnal respiratory polygraphy and body composition measurements were taken with bioelectrical impedance analysis. OPG serum levels were measured using the enzyme-linked immunosorbent assay (ELISA) method. In OSA patients OPG correlated negatively with muscle mass percentage (MM%), phase angle, fat free mass percentage (FFM%) and body cell mass percentage (BCM%), while there was a positive correlation between osteoprotegerin and fat mass percentage (FM%). We demonstrated higher OPG serum levels in OSA patients with cardiovascular diseases than in those without comorbidities (4.01 vs 3.46pmol/l, p<0.05). Our findings, combined with previous observations in other diseases, suggest that elevated OPG serum levels together with selected body composition parameters may be helpful in identifying OSA patients with increased cardiovascular risk.


Comparative proteomics of symbiotic and aposymbiotic juvenile soft corals.
The symbiotic association between corals and photosynthetic unicellular algae is of great importance in coral reef ecosystems. The study of symbiotic relationships is multidisciplinary and involves research in phylogeny, physiology, biochemistry, and ecology. An intriguing phase in each symbiotic relationship is its initiation, in which the partners interact for the first time. The examination of this phase in coral-algae symbiosis from a molecular point of view is still at an early stage. In the present study we used 2-dimensional polyacrylamide gel electrophoresis to compare patterns of proteins synthesized in symbiotic and aposymbiotic primary polyps of the Red Sea soft coral Heteroxenia fuscescens. This is the first work to search for symbiosis-specific proteins during the natural onset of symbiosis in early host ontogeny. The protein profiles reveal changes in the host soft coral proteome through development, but surprisingly virtually no changes in the host proteome as a function of symbiotic state.


Brief report: depression and history of suicide attempts in adults with new-onset Type 2 Diabetes.
To assess past suicide attempts in a cohort of adults with Type 2 Diabetes diagnosed within the prior 24 months. Outpatients were recruited from diabetes education classes or diabetes shared medical appointment. Participants aged 18 or over with a self-reported diagnosis of Type 2 Diabetes (T2DM) in the prior 24 months completed questionnaires about medical (including diabetes), psychiatric, and social history. Participants also completed two screening questionnaires for depression: Patient Health Questionnaire 9 and the Questionnaire Inventory for Depressive Symptoms-Self Report. Those who screened positive for depression had confirmatory testing with a clinician administered Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) checklist. In this convenience sample of 145 patients with Type 2 Diabetes, 9.7% of patients had history of a suicide attempt and 38.2% met diagnosis for major depressive disorder (MDD). Patients with MDD were more likely to have a history of suicide attempts than those without MDD (p=0.0002). Of the patients with prior suicide attempts, 50% screened positive for MDD at the time of the survey. In patients with newly-diagnosed Type 2 Diabetes the rate of past suicide attempts was nearly 10%, which is twice the rate seen in the general population. The rate of past suicide attempts in currently depressed patients with diabetes is 21.8%. These findings suggest the need for monitoring patients with diabetes and depression for future suicide risk.