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Sample records for hyperinsulinemic euglycemic clamp

  1. Evaluation of insulin sensitivity in hyperprolactinemic subjects by euglycemic hyperinsulinemic clamp technique.

    Science.gov (United States)

    Tuzcu, Alpaslan; Yalaki, Serkan; Arikan, Senay; Gokalp, Deniz; Bahcec, Mithat; Tuzcu, Sadiye

    2009-01-01

    The background and aim of the study is to evaluate insulin sensitivity in hyperprolactinemic subjects via euglycemic hyperinsulinemic clamp technique. Sixteen hyperprolactinemic subjects and 12 healthy subjects were included in the study. HOMA-B and HOMA-IR values of groups were calculated. Euglycemic hyperinsulinemic clamp technique was performed in both groups, and the M value of the groups was defined. Mann-Whitney U and chi-square tests were used in statistical analysis. Basal insulin level of hyperprolactinemic patients were higher than the control group (6.85 +/- 4.68; 3.66 +/- 0.88 microU/ml respectively; P < 0.05). Mean HOMA-IR and HOMA-B values of patients were higher than control group (1.49 +/- 1.30; 0.78 +/- 0.27 respectively; P = 0.02 and 136.28 +/- 72.53; 64.77 +/- 23.31, respectively, P < 0.001). M values of the patients were statistically lower than the control group (5.64 +/- 2.36; 7.05 +/- 1.62 kg/mg/min respectively; P < 0.05). (1) Hyperprolactinemic patients were more insulin resistant than control subjects. (2) Insulin resistance in hyperprolactinemic patients is not associated with obesity or anthropometric parameters such as fat content, waist circumference and BMI.

  2. Endotoxemia-induced lymphocyte apoptosis is augmented by a hyperinsulinemic-euglycemic clamp

    DEFF Research Database (Denmark)

    Nielsen, Jeppe Sylvest; A, Larsson; Brix-Christensen, Vibeke

    2005-01-01

    BACKGROUND: Sepsis and endotoxemia are associated with lymphocyte apoptosis. This has been regarded as harmful, contributing to further immune suppression in already immune-compromised patients. Because normalization of blood glucose improves outcome in critically ill patients, the authors...... hypothesized that one of the effects of insulin and normoglycemia would be inhibition of lymphocyte apoptosis. Therefore, in this experimental study in pigs, the authors examined the separate and combined effects of acute endotoxemia and a hyperinsulinemic-euglycemic clamp (HEC) on lymphocyte apoptosis....... METHODS: After 60 min of stabilization, 38 anesthetized and mechanically ventilated pigs (weight, 35-40 kg) were divided (by randomization performed before the experiment) into four groups and were then studied for 570 min. Group 1 received no intervention. Group 2 received a HEC (5 mm p-glucose, insulin...

  3. [Insulin resistance frecuency in women's with polycystic ovary syndrome using hyperinsulinemic-euglycemic clamp].

    Science.gov (United States)

    Vital-Reyes, Victor Saúl; Carrillo-Martínez, Cristo Rey; Hinojosa-Cruz, Juan Carlos; Martínez-Basilia, Azucena; López-Alarcón, Mardya

    2014-12-01

    To assess IR in PCOS patients, using the HE-clamp as the IR gold standard. A transversal design was done. All patients who accepted to participate provided written informed consent. PCOS was diagnosed according to the 2003 Rotterdam criteria. IR was assessed using the H-clamp, and other IR surrogates such as; fasting plasma insulin (FPI), homeostasis model assessment (HOMA) index and insulin/ glucose rate (I/G rate). Statistical analysis was performed using measures of location and spread was used according to data distribution. 21 patients were included. The mean age of the total group studied was 29.5 +/- 4.8 years, and the body mass index (BMI) was 27.2 +/- 3.08 kg/m2. The 85.7% of the patients met the three Rotterdam criteria for the diagnosis of PCOS. According to the HE clamp 95.2% were IR (M/I value < 6 mg/Kg/min), in contrast the prevalence of IR using sur- rogates was 47.6%, 33.3%, and 66.6% for FPI, G/I rate, and HOMA index respectively. Our findings show that IR is highly prevalent in patients with PCOS, and that this prevalence is even higher when insulin sensitivity is assessed using the glucose clamp technique. This evidence suggests that IR could be considered diagnostic criteria for PCOS.

  4. Glucose and fat metabolism in narcolepsy and the effect of sodium oxybate: a hyperinsulinemic-euglycemic clamp study.

    Science.gov (United States)

    Donjacour, Claire E H M; Aziz, N Ahmad; Overeem, Sebastiaan; Kalsbeek, Andries; Pijl, Hanno; Lammers, Gert Jan

    2014-04-01

    Narcolepsy is associated with obesity though it is uncertain whether this is caused by changes in glucose and fat metabolism. Therefore, we performed a detailed analysis of systemic energy homeostasis in narcolepsy patients, and additionally, investigated whether it was affected by three months of sodium oxybate (SXB) treatment. Nine hypocretin deficient patients with narcolepsy-cataplexy, and nine healthy sex, age, and BMI matched controls were enrolled. A hyperinsulinemic-euglycemic clamp combined with stable isotopes ([6,6-(2)H2]-glucose and [(2)H5]- glycerol) was performed at baseline. In seven patients a second study was performed after three months of SXB treatment. Glucose disposal rate (GDR) per unit serum insulin was significantly higher in narcolepsy patients compared to matched controls (1.6 ± 0.2 vs. 1.1 ± 0.3 μmol/kgFFM/min/mU×L; P = 0.024), whereas β-cell function was similar (P = 0.50). Basal steady state glycerol appearance rate tended to be lower in narcolepsy patients (5.2 ± 0.4 vs. 7.5 ± 1.3 μmol/kgFM/min; P = 0.058), suggesting a lower rate of lipolysis. SXB treatment induced a trend in reduction of the GDR (1.4 ± 0.1 vs. 1.1 ± 0.2 μmol/kgFFM/min/mU×L; P = 0.063) and a reduction in endogenous glucose production (0.24 ± 0.03 vs. 0.16 ± 0.03 μmol/kgFFM/min/mU×L: P = 0.028) per unit serum insulin. After SXB treatment lipolysis increased (4.9 ± 0.4 vs. 6.5 ± 0.6 μmol/kgFM/min; P = 0.018), and body weight decreased in narcolepsy patients (99.2 ± 6.0 vs. 94.0 ± 5.4 kg; P = 0.044). We show that narcolepsy patients are more insulin sensitive and may have a lower rate of lipolysis than matched controls. SXB stimulated lipolysis in narcolepsy patients, possibly accounting for the weight loss after treatment. While sodium oxybate tended to decrease systemic insulin sensitivity, it increased hepatic insulin sensitivity, suggesting tissue-specific effects.

  5. Elevation of serum insulin concentration during euglycemic hyperinsulinemic clamp studies leads to similar activation of insulin receptor kinase in skeletal muscle of subjects with and without NIDDM

    DEFF Research Database (Denmark)

    Klein, H H; Vestergaard, H; Kotzke, G

    1995-01-01

    The role of skeletal muscle insulin receptor kinase in the pathogenesis of non-insulin-dependent diabetes mellitus (NIDDM) was investigated. Muscle biopsies from 13 patients with NIDDM and 10 control subjects at fasting serum insulin concentrations and approximately 1,000 pmol/l steady-state serum...... insulin during euglycemic hyperinsulinemic clamps were immediately frozen. The biopsies were then solubilized, and the receptors were immobilized to anti-insulin receptor antibody-coated microwells. Receptor kinase and binding activities were consecutively measured in these wells. The increase in serum...... insulin concentration (73 +/- 14 to 1,004 +/- 83 and 45 +/- 7 to 1,07 +/- 77 pmol/l in the NIDDM and control groups, respectively) had similar effects on receptor kinase activity in both study groups (12 +/- 1 to 42 +/- 5 and 12 +/- 2 to 47 +/- 5 amol P.fmol binding activity-1. min-1 in the NIDDM...

  6. Low-carbohydrate diet versus euglycemic hyperinsulinemic clamp for the assessment of myocardial viability with 18F-fluorodeoxyglucose-PET: a pilot study.

    Science.gov (United States)

    Soares, José; Rodrigues Filho, Filadelfo; Izaki, Marisa; Giorgi, Maria Clementina P; Catapirra, Rosa M A; Abe, Rubens; Vinagre, Carmen G C M; Cerri, Giovanni G; Meneghetti, José Cláudio

    2014-02-01

    Positron emission tomography with (18)F-fluorodeoxyglucose (FDG-PET) is considered the gold standard for myocardial viability. A pilot study was undertaken to compare FDG-PET using euglycemic hyperinsulinemic clamp before (18)F-fluorodeoxyglucose ((18)F-FDG) administration (PET-CLAMP) with a new proposed technique consisting of a 24-h low-carbohydrate diet before (18)F-FDG injection (PET-DIET), for the assessment of hypoperfused but viable myocardium (hibernating myocardium). Thirty patients with previous myocardial infarction were subjected to rest (99m)Tc-sestamibi-SPECT and two (18)F-FDG studies (PET-CLAMP and PET-DIET). Myocardial tracer uptake was visually scored using a 5-point scale in a 17-segment model. Hibernating myocardium was defined as normal or mildly reduced metabolism ((18)F-FDG uptake) in areas with reduced perfusion ((99m)Tc-sestamibi uptake) since (18)F-FDG uptake was higher than the degree of hypoperfusion-perfusion/metabolism mismatch indicating a larger flow defect. PET-DIET identified 79 segments and PET-CLAMP 71 as hibernating myocardium. Both methods agreed in 61 segments (agreement = 94.5 %, κ = 0.78). PET-DIET identified 230 segments and PET-CLAMP 238 as nonviable. None of the patients had hypoglycemia after DIET, while 20 % had it during CLAMP. PET-DIET compared with PET-CLAMP had a good correlation for the assessment of hibernating myocardium. To our knowledge, these data provide the first evidence of the possibility of myocardial viability assessment with this technique.

  7. The GLP-1 Analogue Exenatide Improves Hepatic and Muscle Insulin Sensitivity in Diabetic Rats: Tracer Studies in the Basal State and during Hyperinsulinemic-Euglycemic Clamp

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    Hui Wu

    2014-01-01

    Full Text Available Objective. Glucagon-like peptide-1 (GLP-1 analogues (e.g., exenatide increase insulin secretion in diabetes but less is known about their effects on glucose production or insulin-stimulated glucose uptake in peripheral tissues. Methods. Four groups of Sprague-Dawley rats were studied: nondiabetic (control, C; nondiabetic + exenatide (C + E; diabetic (D; diabetic + exenatide (D + E with diabetes induced by streptozotocin and high fat diet. Infusion of 3-3H-glucose and U-13C-glycerol was used to measure basal rates of appearance (Ra of glucose and glycerol and gluconeogenesis from glycerol (GNG. During hyperinsulinemic-euglycemic clamp, glucose uptake into gastrocnemius muscles was measured with 2-deoxy-D-14C-glucose. Results. In the diabetic rats, exenatide reduced the basal Ra of glucose (P<0.01 and glycerol (P<0.01 and GNG (P<0.001. During the clamp, Ra of glucose was also reduced, whereas the rate of disappearance of glucose increased and there was increased glucose uptake into muscle (P<0.01 during the clamp. In the nondiabetic rats, exenatide had no effect. Conclusion. In addition to its known effects on insulin secretion, administration of the GLP-1 analogue, exenatide, is associated with increased inhibition of gluconeogenesis and improved glucose uptake into muscle in diabetic rats, implying improved hepatic and peripheral insulin sensitivity.

  8. Accuracy and Time Delay of Glucose Measurements of Continuous Glucose Monitoring and Bedside Artificial Pancreas During Hyperglycemic and Euglycemic Hyperinsulinemic Glucose Clamp Study.

    Science.gov (United States)

    Kuroda, Akio; Taniguchi, Satoshi; Akehi, Yuko; Mori, Hiroyasu; Tamaki, Motoyuki; Suzuki, Reiko; Otsuka, Yinhua; Matsuhisa, Munehide

    2017-11-01

    Glucose values of continuous glucose monitoring (CGM) have time delays compared with plasma glucose (PG) values. The artificial pancreas (STG-55, Nikkiso, Japan) (AP), which measures venous blood glucose directly, also has a time delay because of the long tubing lines from sampling vessel to the glucose sensor. We investigate accuracy and time delay of CGM and AP in comparison with PG values during 2-step glucose clamp study. Seven patients with type 2 diabetes and 2 healthy volunteers were included in this study. CGM (Enlite sensor, Medtronic, Northridge, CA, USA) was attached on the day before the experiment. Hyperglycemic (200 mg/dL) clamp was performed for 90 minutes, followed by euglycemic (100 mg/dL) hyperinsulinemic (100 μU/mL) clamp for 90-120 minutes using AP. CGM sensor glucose was calibrated just before and after the clamp study. AP and CGM values were compared with PG values. AP values were significantly lower than PG values at 5, 30 minute during hyperglycemic clamp. In comparison, CGM value at 0 minute was significantly higher, and its following values were almost significantly lower than PG values. The time delay of AP and CGM values to reach maximum glucose levels were 5.0 ± 22.3 (NS) and 28.6 ± 32.5 ( P < .05) min, respectively. Mean absolute rate difference of CGM was significantly higher than AP (24.0 ± 7.6 vs 15.3 ± 4.6, P < .05) during glucose rising period (0-45 min); however, there were no significant differences during other periods. Both CGM and AP failed to follow plasma glucose values during nonphysiologically rapid glucose rising, but indicated accurate values during physiological glucose change.

  9. Use of the hyperinsulinemic euglycemic clamp to assess insulin sensitivity in guinea pigs: dose response, partitioned glucose metabolism, and species comparisons.

    Science.gov (United States)

    Horton, Dane M; Saint, David A; Owens, Julie A; Gatford, Kathryn L; Kind, Karen L

    2017-07-01

    The guinea pig is an alternate small animal model for the study of metabolism, including insulin sensitivity. However, only one study to date has reported the use of the hyperinsulinemic euglycemic clamp in anesthetized animals in this species, and the dose response has not been reported. We therefore characterized the dose-response curve for whole body glucose uptake using recombinant human insulin in the adult guinea pig. Interspecies comparisons with published data showed species differences in maximal whole body responses (guinea pig ≈ human guinea pig > human ≈ rat > mouse). In subsequent studies, we used concomitant d-[3-3H]glucose infusion to characterize insulin sensitivities of whole body glucose uptake, utilization, production, storage, and glycolysis in young adult guinea pigs at human insulin doses that produced approximately half-maximal (7.5 mU·min-1·kg-1) and near-maximal whole body responses (30 mU·min-1·kg-1). Although human insulin infusion increased rates of glucose utilization (up to 68%) and storage and, at high concentrations, increased rates of glycolysis in females, glucose production was only partially suppressed (~23%), even at high insulin doses. Fasting glucose, metabolic clearance of insulin, and rates of glucose utilization, storage, and production during insulin stimulation were higher in female than in male guinea pigs (P guinea pigs, allowing studies of regulation of insulin sensitivity in this species. Copyright © 2017 the American Physiological Society.

  10. Validation of different measures of insulin sensitivity of glucose metabolism in dairy cows using the hyperinsulinemic euglycemic clamp test as the gold standard.

    Science.gov (United States)

    De Koster, J; Hostens, M; Hermans, K; Van den Broeck, W; Opsomer, G

    2016-10-01

    The aim of the present research was to compare different measures of insulin sensitivity in dairy cows at the end of the dry period. To do so, 10 clinically healthy dairy cows with a varying body condition score were selected. By performing hyperinsulinemic euglycemic clamp (HEC) tests, we previously demonstrated a negative association between the insulin sensitivity and insulin responsiveness of glucose metabolism and the body condition score of these animals. In the same animals, other measures of insulin sensitivity were determined and the correlation with the HEC test, which is considered as the gold standard, was calculated. Measures derived from the intravenous glucose tolerance test (IVGTT) are based on the disappearance of glucose after an intravenous glucose bolus. Glucose concentrations during the IVGTT were used to calculate the area under the curve of glucose and the clearance rate of glucose. In addition, glucose and insulin data from the IVGTT were fitted in the minimal model to derive the insulin sensitivity parameter, Si. Based on blood samples taken before the start of the IVGTT, basal concentrations of glucose, insulin, NEFA, and β-hydroxybutyrate were determined and used to calculate surrogate indices for insulin sensitivity, such as the homeostasis model of insulin resistance, the quantitative insulin sensitivity check index, the revised quantitative insulin sensitivity check index and the revised quantitative insulin sensitivity check index including β-hydroxybutyrate. Correlation analysis revealed no association between the results obtained by the HEC test and any of the surrogate indices for insulin sensitivity. For the measures derived from the IVGTT, the area under the curve for the first 60 min of the test and the Si derived from the minimal model demonstrated good correlation with the gold standard. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Using the hyperinsulinemic euglycemic clamp to assess insulin sensitivity at 3 months following Roux-en-Y gastric bypass surgery in type 2 diabetes patients with BMI <35 kg/m2 in China.

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    Zhao, Lei; Zhu, Liyong; Su, Zhihong; Liu, Yong; Li, Pengzhou; Yang, Xiangwu; Li, Weizheng; Tan, Lingjie; Sun, Xulong; Zhu, Shaihong

    2017-02-01

    The aim of this study was to describe short-time effect of Roux-en-Y gastric bypass (RYGB) on insulin sensitivity (IS) of Chinese diabetes patients with body mass index (BMI) clamp. We studied 15 type 2 diabete mellitus (T2DM) patients with BMI clamp were performed at baseline and at 3 months after LRYGB. The glucose disposal rate (M value) increased significantly at 3 months after RYGB (from 3.36 ± 1.26 mg kg-1 min-1 to 6.30 ± 1.3 mg kg-1 min-1, p clamp reduced remarkably from baseline to 3 months after RYGB (from 114.40 ± 6.11 min to 97.93 ± 8.57 min, p clamp may be an useful index of assessing insulin sensitivity. It is more reasonal to combine the time to reach euglycemia with M value for assessing IS. Copyright © 2016 IJS Publishing Group Ltd. Published by Elsevier Ltd. All rights reserved.

  12. Triglyceride glucose index as a surrogate measure of insulin sensitivity in obese adolescents with normoglycemia, prediabetes, and type 2 diabetes mellitus: Comparison with the hyperinsulinemic–euglycemic clamp

    Science.gov (United States)

    There is a need for simple surrogate estimates of insulin sensitivity in epidemiological studies of obese youth because the hyperinsulinemic-euglycemic clamp is not feasible on a large scale. Objectives: (i) To examine the triglyceride glucose (TyG) index (Ln[fasting triglycerides (mg/dL)'×'fasting ...

  13. Hyperinsulinemic clamp modulates milk fat globule lipid composition in goats.

    Science.gov (United States)

    Argov-Argaman, N; Mbogori, T; Sabastian, C; Shamay, A; Mabjeesh, S J

    2012-10-01

    We determined the effect of insulin on milk fatty acid (FA) and lipid composition in goats. Four dairy goats, 150 d in milk, were subjected to hyperinsulinemic clamp (treatment) or saline (control) infusion for 4d in a crossover design study. Composition and concentration of plasma and milk FA, triglycerides, phospholipids, sphingolipids, and cholesterol were determined. Mammary gland biopsies were taken at the end of each experimental period and lipogenic gene expression was determined. Plasma insulin was elevated 3.5-fold, whereas plasma glucose remained constant during the treatment period. Feed intake decreased by 26% and fat yield decreased by 17% relative to controls. No change in nonesterified FA concentration was found between controls and treatment. Compared with controls, insulin decreased yield of long-chain saturated FA by 14%. Milk concentration of long-chain FA was reduced by 3%, whereas that of medium-chain FA increased by 5% during the treatment compared with controls. Hyperinsulinemic clamps increased the yields of milk phospholipids by 9% and cholesterol by 16%, whereas it only tended to decrease triglyceride yields (by 11%). Hyperinsulinemic treatment resulted in compositional changes in the milk fat globule membrane, as reflected by 15 and 9% decreases in phosphatidylethanolamine and phosphatidylcholine concentrations, respectively. Lipogenic gene expression of acyl coenzyme A carboxylase, stearoyl coenzyme A desaturase, and FA synthase did not change, whereas lipoprotein lipase gene expression tended toward an increase in the treatment period compared with controls. Hyperinsulinemic clamps reduce the availability of long-chain FA, which are considered to originate from the diet and adipose lipolysis for milk lipid synthesis by the mammary gland of goats. Under these conditions, long-chain FA might be preferentially channeled to phospholipid rather than triglyceride synthesis, hence increasing phospholipid yields. Mechanisms determining FA

  14. Influence of non-steady state during isoglycemic hyperinsulinemic clamp in hypertension. A LIFE substudy

    DEFF Research Database (Denmark)

    Olsen, M H; Andersen, U B; Wachtell, K

    1999-01-01

    We wanted to investigate whether time to steady state was reached within 2 h of insulin infusion during isoglycemic hyperinsulinemic clamp, comparing the glucose uptake index (M/IG) with Bergman's insulin sensitivity index (Sip). We performed a 2-h oral glucose tolerance test and a 3-h isoglycemic...... hyperinsulinemic clamp in 26 young, healthy subjects and 43 elderly patients with unmedicated essential hypertension and left ventricular hypertrophy. The 3-h Sip correlated strongly with the 2-h M/IG in the patients (r = 0.88, p .... Because the 2-h M/IG correlated strongly with the 3-h Sip with relatively narrow limits of agreement, it is a good measure of insulin sensitivity. However, a 2-h clamp results in lower insulin sensitivity values in elderly, hypertensive patients due to the fact that steady state is not reached...

  15. Influence of non-steady state during isoglycemic hyperinsulinemic clamp in hypertension. A LIFE substudy

    DEFF Research Database (Denmark)

    Olsen, M H; Andersen, U B; Wachtell, K

    1999-01-01

    We wanted to investigate whether time to steady state was reached within 2 h of insulin infusion during isoglycemic hyperinsulinemic clamp, comparing the glucose uptake index (M/IG) with Bergman's insulin sensitivity index (Sip). We performed a 2-h oral glucose tolerance test and a 3-h isoglycemic...... hyperinsulinemic clamp in 26 young, healthy subjects and 43 elderly patients with unmedicated essential hypertension and left ventricular hypertrophy. The 3-h Sip correlated strongly with the 2-h M/IG in the patients (r = 0.88, p ... narrow limits of agreement in the patients. However, during the third hour of insulin infusion, M/IG (10.0 vs 12.21(2) x kg(-1) x min(-1) x mmol(-1), p Sip (7.1 vs 9.41(2) x kg(-1) x min(-1) x mmol(-1), p

  16. Hyperinsulinemic-normoglycemic clamp administered together with amino acids induces anabolism after cardiac surgery.

    Science.gov (United States)

    Codère-Maruyama, Takumi; Schricker, Thomas; Shum-Tim, Dominique; Wykes, Linda; Nitschmann, Evan; Guichon, Céline; Kristof, Arnold S; Hatzakorzian, Roupen

    2016-12-01

    Cardiac surgery triggers an inflammatory stress response, leading to protein catabolism, a process that even high-dose insulin therapy alone cannot reverse. To determine whether hyperinsulinemic-normoglycemic clamp and perioperative amino acid (AA) supplementation improves whole body protein balance, 20 patients scheduled for elective coronary artery bypass grafting surgery were randomly assigned to have intra- and postoperative hyperinsulinemic-normoglycemic clamp, with or without intravenous AA supplementation. Primed continuous infusions of [6,6-2H2]glucose and l-[1-13C]leucine were used to quantify whole body protein and glucose metabolism before and after surgery. Adipose tissue and serum cytokines were also analyzed to measure their responsiveness to the anabolic effect of AA administration. During hyperinsulinemic-normoglycemic clamp, AA supplementation successfully stimulated whole body protein synthesis, resulting in a positive whole body protein balance after surgery (insulin: -13.6 ± 4.5 vs. insulin + AA: 2.1 ± 5.4 μmol·kg-1·h-1, P anabolic state after open-heart surgery, as quantified by a positive whole body protein balance. Copyright © 2016 the American Physiological Society.

  17. Measuring beta-cell function relative to insulin sensitivity in youth: Does the hyperglycemic clamp suffice?

    Science.gov (United States)

    To compare beta-cell function relative to insulin sensitivity, disposition index (DI), calculated from two clamps (2cDI, insulin sensitivity from the hyperinsulinemic-euglycemic clamp and first-phase insulin from the hyperglycemic clamp) with the DI calculated from the hyperglycemic clamp alone (hcD...

  18. Glucose and fat metabolism in narcolepsy and the effect of sodium oxybate: a hyperinsulinemic-euglycemic clamp study

    NARCIS (Netherlands)

    Donjacour, Claire E. H. M.; Aziz, N. Ahmad; Overeem, Sebastiaan; Kalsbeek, Andries; Pijl, Hanno; Lammers, Gert Jan

    2014-01-01

    Narcolepsy is associated with obesity though it is uncertain whether this is caused by changes in glucose and fat metabolism. Therefore, we performed a detailed analysis of systemic energy homeostasis in narcolepsy patients, and additionally, investigated whether it was affected by three months of

  19. Glucose and fat metabolism in narcolepsy and the effect of sodium oxybate : a hyperinsulinemic-euglycemic clamp study

    NARCIS (Netherlands)

    Donjacour, Claire E H M; Aziz, N Ahmad; Overeem, Sebastiaan; Kalsbeek, A.; Pijl, Hanno; Lammers, Gert Jan

    2014-01-01

    INTRODUCTION: Narcolepsy is associated with obesity though it is uncertain whether this is caused by changes in glucose and fat metabolism. Therefore, we performed a detailed analysis of systemic energy homeostasis in narcolepsy patients, and additionally, investigated whether it was affected by

  20. Glucose and fat metabolism in narcolepsy and the effect of sodium oxybate: a hyperinsulinemic-euglycemic clamp study

    NARCIS (Netherlands)

    Donjacour, C.E.; Aziz, N.A.; Overeem, S.; Kalsbeek, A.; Pijl, H.; Lammers, G.J.

    2014-01-01

    INTRODUCTION: Narcolepsy is associated with obesity though it is uncertain whether this is caused by changes in glucose and fat metabolism. Therefore, we performed a detailed analysis of systemic energy homeostasis in narcolepsy patients, and additionally, investigated whether it was affected by

  1. Effects of the rate of insulin infusion during isoglycemic, hyperinsulinemic clamp procedures on measures of insulin action in healthy, mature thoroughbred mares.

    Science.gov (United States)

    Urschel, K L; Escobar, J; McCutcheon, L J; Geor, R J

    2014-04-01

    The objective of this study was to determine whether the rate of insulin infusion during isoglycemic hyperinsulinemic clamp procedures affected measures of insulin action, including glucose disposal and plasma non-esterified fatty acid, endothelin-1, and nitric oxide concentrations, in mature, healthy horses. Eight thoroughbred mares were studied during a 2-h hyperinsulinemic clamp procedure, conducted at each of 4 rates of insulin infusion: 0 (CON), 1.2 (LOWINS), 3 (MEDINS), and 6 (HIGHINS) mU · kg(-1) · min(-1). The infusion rate of a dextrose solution was adjusted throughout the clamp procedures to maintain blood glucose levels within 10% of baseline glucose concentrations. Plasma insulin concentrations were measured throughout the clamp procedures, and used with the rate of glucose infusion to calculate the plasma insulin concentration-to-rate of glucose infusion ratio, a measure of insulin action on glucose disposal. The rate of glucose infusion increased with rate of insulin infusion (P clamp procedures in the LOWINS, MEDINS, and HIGHINS treatments (P 0.05). The data indicate that it is important to standardize insulin infusion rate if data are to be compared between hyperinsulinemic clamp studies. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. In vivo cardiac glucose metabolism in the high-fat fed mouse: Comparison of euglycemic–hyperinsulinemic clamp derived measures of glucose uptake with a dynamic metabolomic flux profiling approach

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    Kowalski, Greg M., E-mail: greg.kowalski@deakin.edu.au [Centre for Physical Activity and Nutrition Research, School of Exercise and Nutrition Sciences, Deakin University, Burwood, Victoria 3125 (Australia); De Souza, David P. [Metabolomics Australia, Department of Biochemistry and Molecular Biology, Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Parkville, Victoria 3010 (Australia); Risis, Steve [Cellular and Molecular Metabolism Laboratory, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria 3004 (Australia); Burch, Micah L. [Brigham and Women' s Hospital, Department of Medicine, Boston, MA (United States); Hamley, Steven [Centre for Physical Activity and Nutrition Research, School of Exercise and Nutrition Sciences, Deakin University, Burwood, Victoria 3125 (Australia); Kloehn, Joachim [Metabolomics Australia, Department of Biochemistry and Molecular Biology, Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Parkville, Victoria 3010 (Australia); Selathurai, Ahrathy [Centre for Physical Activity and Nutrition Research, School of Exercise and Nutrition Sciences, Deakin University, Burwood, Victoria 3125 (Australia); Lee-Young, Robert S. [Cellular and Molecular Metabolism Laboratory, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria 3004 (Australia); Tull, Dedreia; O' Callaghan, Sean; McConville, Malcolm J. [Metabolomics Australia, Department of Biochemistry and Molecular Biology, Bio21 Institute of Molecular Science and Biotechnology, University of Melbourne, Parkville, Victoria 3010 (Australia); Bruce, Clinton R. [Centre for Physical Activity and Nutrition Research, School of Exercise and Nutrition Sciences, Deakin University, Burwood, Victoria 3125 (Australia)

    2015-08-07

    Rationale: Cardiac metabolism is thought to be altered in insulin resistance and type 2 diabetes (T2D). Our understanding of the regulation of cardiac substrate metabolism and insulin sensitivity has largely been derived from ex vivo preparations which are not subject to the same metabolic regulation as in the intact heart in vivo. Studies are therefore required to examine in vivo cardiac glucose metabolism under physiologically relevant conditions. Objective: To determine the temporal pattern of the development of cardiac insulin resistance and to compare with dynamic approaches to interrogate cardiac glucose and intermediary metabolism in vivo. Methods and results: Studies were conducted to determine the evolution of cardiac insulin resistance in C57Bl/6 mice fed a high-fat diet (HFD) for between 1 and 16 weeks. Dynamic in vivo cardiac glucose metabolism was determined following oral administration of [U-{sup 13}C] glucose. Hearts were collected after 15 and 60 min and flux profiling was determined by measuring {sup 13}C mass isotopomers in glycolytic and tricarboxylic acid (TCA) cycle intermediates. Cardiac insulin resistance, determined by euglycemic–hyperinsulinemic clamp, was evident after 3 weeks of HFD. Despite the presence of insulin resistance, in vivo cardiac glucose metabolism following oral glucose administration was not compromised in HFD mice. This contrasts our recent findings in skeletal muscle, where TCA cycle activity was reduced in mice fed a HFD. Similar to our report in muscle, glucose derived pyruvate entry into the TCA cycle in the heart was almost exclusively via pyruvate dehydrogenase, with pyruvate carboxylase mediated anaplerosis being negligible after oral glucose administration. Conclusions: Under experimental conditions which closely mimic the postprandial state, the insulin resistant mouse heart retains the ability to stimulate glucose metabolism. - Highlights: • Insulin clamp was used to determine the evolution of cardiac

  3. Fed-state clamp stimulates cellular mechanisms of muscle protein anabolism and modulates glucose disposal in normal men.

    Science.gov (United States)

    Adegoke, Olasunkanmi A J; Chevalier, Stéphanie; Morais, José A; Gougeon, Réjeanne; Kimball, Scot R; Jefferson, Leonard S; Wing, Simon S; Marliss, Errol B

    2009-01-01

    Since maximum anabolism occurs postprandially, we developed a simulated fed state with clamped hyperinsulinemia, physiological hyperglycemia, and hyperaminoacidemia (Hyper-3) and explored muscle cellular mechanisms. Whole body [1-(13)C]leucine and [3-(3)H]glucose kinetics in healthy men were compared between hyperinsulinemic, euglycemic, isoaminoacidemic (Hyper-1, n = 10) and Hyper-3 (n = 9) clamps. In Hyper-3 vs. Hyper-1, nonoxidative leucine R(d) [rate of disappearance (synthesis)] was stimulated more (45 +/- 4 vs. 24 +/- 4 micromol/min, P anabolism, and muscle protein translation initiation pathways and decreases protein ubiquitination. The main contribution of hyperaminoacidemia is stimulation of synthesis rather than inhibition of proteolysis, and it attenuates the expected increment of glucose disposal.

  4. Computer Simulation Model to Train Medical Personnel on Glucose Clamp Procedures.

    Science.gov (United States)

    Maghoul, Pooya; Boulet, Benoit; Tardif, Annie; Haidar, Ahmad

    2017-10-01

    A glucose clamp procedure is the most reliable way to quantify insulin pharmacokinetics and pharmacodynamics, but skilled and trained research personnel are required to frequently adjust the glucose infusion rate. A computer environment that simulates glucose clamp experiments can be used for efficient personnel training and development and testing of algorithms for automated glucose clamps. We built 17 virtual healthy subjects (mean age, 25±6 years; mean body mass index, 22.2±3 kg/m2), each comprising a mathematical model of glucose regulation and a unique set of parameters. Each virtual subject simulates plasma glucose and insulin concentrations in response to intravenous insulin and glucose infusions. Each virtual subject provides a unique response, and its parameters were estimated from combined intravenous glucose tolerance test-hyperinsulinemic-euglycemic clamp data using the Bayesian approach. The virtual subjects were validated by comparing their simulated predictions against data from 12 healthy individuals who underwent a hyperglycemic glucose clamp procedure. Plasma glucose and insulin concentrations were predicted by the virtual subjects in response to glucose infusions determined by a trained research staff performing a simulated hyperglycemic clamp experiment. The total amount of glucose infusion was indifferent between the simulated and the real subjects (85±18 g vs. 83±23 g; p=NS) as well as plasma insulin levels (63±20 mU/L vs. 58±16 mU/L; p=NS). The virtual subjects can reliably predict glucose needs and plasma insulin profiles during hyperglycemic glucose clamp conditions. These virtual subjects can be used to train personnel to make glucose infusion adjustments during clamp experiments. Copyright © 2017 Diabetes Canada. Published by Elsevier Inc. All rights reserved.

  5. Euglycemic clamp insulin sensitivity and longitudinal systolic blood pressure

    DEFF Research Database (Denmark)

    Petrie, John R; Malik, Muhammad Omar; Balkau, Beverley

    2013-01-01

    Insulin resistance may be an independent risk factor for the development of hypertension, but change in blood pressure (BP) over time has not been adequately studied in healthy individuals fully characterized for insulin sensitivity. In the Relationship between Insulin Sensitivity...

  6. Substrate metabolism during basal and hyperinsulinemic conditions in adolescents and young-adults with Barth syndrome.

    Science.gov (United States)

    Cade, W Todd; Spencer, Carolyn T; Reeds, Dominic N; Waggoner, Alan D; O'Connor, Robert; Maisenbacher, Melissa; Crowley, Jan R; Byrne, Barry J; Peterson, Linda R

    2013-01-01

    Barth syndrome (BTHS) is a rare X-linked disorder that is characterized by mitochondrial abnormalities, infantile or childhood onset of cardioskeletal myopathy, and high mortality rates. It is currently unknown if BTHS related mitochondrial dysfunction results in substrate metabolism abnormalities and thereby contributes to cardioskeletal myopathy in patients with BTHS. Adolescents and young adults with BTHS (n = 5, 20 ± 4 yrs) and age and activity matched healthy controls (n = 5, 18 ± 4 yrs) underwent an hyperinsulinemic-euglycemic clamp procedure with stable isotopically labeled tracers for measurement of lipolysis, fatty acid oxidation, glucose disposal, and whole-body proteolysis rates; dual energy x-ray absorptiometry for measurement of body composition and 2-D and strain echocardiography for measurement of left ventricular function. Participants with BTHS had lower fat-free mass (FFM) (BTHS: 31.4 ± 6.9 vs. 46.7 ± 5.3 kg, p trend towards higher basal leucine rate of appearance per kg FFM (BTHS: 271.4 ± 69.3 vs. 193.1 ± 28.7 μmol/kgFFM/hr, p = 0.07) compared to controls. Higher basal leucine rate of appearance per kg FFM (i.e. whole-body proteolytic rate) tended to be associated with lower left ventricular systolic strain (r = -0.57, p = 0.09). Whole-body fatty acid, glucose and amino acid metabolism kinetics when expressed per unit of body composition are altered and appear to be related to cardioskeletal myopathy in humans with BTHS. Further studies examining myocardial substrate metabolism and whole-body substrate metabolism during increased energy demands (e.g., exercise) and their relationships to skeletal and cardiac function are recommended.

  7. Artificial neural networks based controller for glucose monitoring during clamp test.

    Directory of Open Access Journals (Sweden)

    Merav Catalogna

    Full Text Available Insulin resistance (IR is one of the most widespread health problems in modern times. The gold standard for quantification of IR is the hyperinsulinemic-euglycemic glucose clamp technique. During the test, a regulated glucose infusion is delivered intravenously to maintain a constant blood glucose concentration. Current control algorithms for regulating this glucose infusion are based on feedback control. These models require frequent sampling of blood, and can only partly capture the complexity associated with regulation of glucose. Here we present an improved clamp control algorithm which is motivated by the stochastic nature of glucose kinetics, while using the minimal need in blood samples required for evaluation of IR. A glucose pump control algorithm, based on artificial neural networks model was developed. The system was trained with a data base collected from 62 rat model experiments, using a back-propagation Levenberg-Marquardt optimization. Genetic algorithm was used to optimize network topology and learning features. The predictive value of the proposed algorithm during the temporal period of interest was significantly improved relative to a feedback control applied at an equivalent low sampling interval. Robustness to noise analysis demonstrates the applicability of the algorithm in realistic situations.

  8. Post clamp

    Science.gov (United States)

    Ramsey, John K. (Inventor); Meyn, Erwin H. (Inventor)

    1990-01-01

    A pair of spaced collars are mounted at right angles on a clamp body by retaining rings which enable the collars to rotate with respect to the clamp body. Mounting posts extend through aligned holes in the collars and clamp body. Each collar can be clamped onto the inserted post while the clamp body remains free to rotate about the post and collar. The clamp body is selectively clamped onto each post.

  9. Postprandial Hyperinsulinemic Hypoglycemia in Bariatric Surgery

    NARCIS (Netherlands)

    de Heide, L. J.M.; Emous, M.; van Beek, A. P.

    2016-01-01

    Postprandial hyperinsulinemic hypoglycemia (PHH) is a complication of bariatric surgery, especially Roux-en-Y gastric bypass. The true incidence is not known as the definition of PHH is not clear. Continuous glucose monitoring shows a prevalence of hypoglycemia in 75% of patients, with only 1 in 5

  10. Replication Clamps and Clamp Loaders

    Science.gov (United States)

    Hedglin, Mark; Kumar, Ravindra; Benkovic, Stephen J.

    2013-01-01

    To achieve the high degree of processivity required for DNA replication, DNA polymerases associate with ring-shaped sliding clamps that encircle the template DNA and slide freely along it. The closed circular structure of sliding clamps necessitates an enzyme-catalyzed mechanism, which not only opens them for assembly and closes them around DNA, but specifically targets them to sites where DNA synthesis is initiated and orients them correctly for replication. Such a feat is performed by multisubunit complexes known as clamp loaders, which use ATP to open sliding clamp rings and place them around the 3′ end of primer–template (PT) junctions. Here we discuss the structure and composition of sliding clamps and clamp loaders from the three domains of life as well as T4 bacteriophage, and provide our current understanding of the clamp-loading process. PMID:23545418

  11. Rigid clamp

    Science.gov (United States)

    Benavides, Gilbert L.; Burt, Jack D.

    1994-01-01

    The invention relates to a clamp mechanism that can be used to attach or temporarily support objects inside of tubular goods. The clamp mechanism can also be modified so that it grips objects. The clamp has a self-centering feature to accommodate out-of-roundness or other internal defections in tubular objects such as pipe. A plurality of clamping shoes are expanded by a linkage which is preferably powered by a motor to contact the inside of a pipe. The motion can be reversed and jaw elements can be connected to the linkage so as to bring the jaws together to grab an object.

  12. BMI, RQ, Diabetes, and Sex Affect the Relationships Between Amino Acids and Clamp Measures of Insulin Action in Humans

    Science.gov (United States)

    Thalacker-Mercer, Anna E.; Ingram, Katherine H.; Guo, Fangjian; Ilkayeva, Olga; Newgard, Christopher B.; Garvey, W. Timothy

    2014-01-01

    Previous studies have used indirect measures of insulin sensitivity to link circulating amino acids with insulin resistance and identify potential biomarkers of diabetes risk. Using direct measures (i.e., hyperinsulinemic-euglycemic clamps), we examined the relationships between the metabolomic amino acid profile and insulin action (i.e., glucose disposal rate [GDR]). Relationships between GDR and serum amino acids were determined among insulin-sensitive, insulin-resistant, and type 2 diabetic (T2DM) individuals. In all subjects, glycine (Gly) had the strongest correlation with GDR (positive association), followed by leucine/isoleucine (Leu/Ile) (negative association). These relationships were dramatically influenced by BMI, the resting respiratory quotient (RQ), T2DM, and sex. Gly had a strong positive correlation with GDR regardless of BMI, RQ, or sex but became nonsignificant in T2DM. In contrast, Leu/Ile was negatively associated with GDR in nonobese and T2DM subjects. Increased resting fat metabolism (i.e., low RQ) and obesity were observed to independently promote and negate the association between Leu/Ile and insulin resistance, respectively. Additionally, the relationship between Leu/Ile and GDR was magnified in T2DM males. Future studies are needed to determine whether Gly has a mechanistic role in glucose homeostasis and whether dietary Gly enrichment may be an effective intervention in diseases characterized by insulin resistance. PMID:24130332

  13. Characterization of the insulin sensitivity of ghrelin receptor KO mice using glycemic clamps

    Directory of Open Access Journals (Sweden)

    Morgan Kristen

    2011-01-01

    Full Text Available Abstract Background We and others have demonstrated previously that ghrelin receptor (GhrR knock out (KO mice fed a high fat diet (HFD have increased insulin sensitivity and metabolic flexibility relative to WT littermates. A striking feature of the HFD-fed GhrR KO mouse is the dramatic decrease in hepatic steatosis. To characterize further the underlying mechanisms of glucose homeostasis in GhrR KO mice, we conducted both hyperglycemic (HG and hyperinsulinemic-euglycemic (HI-E clamps. Additionally, we investigated tissue glucose uptake and specifically examined liver insulin sensitivity. Results Consistent with glucose tolerance-test data, in HG clamp experiments, GhrR KO mice showed a reduction in glucose-stimulated insulin release relative to WT littermates. Nevertheless, a robust 1st phase insulin secretion was still achieved, indicating that a healthy β-cell response is maintained. Additionally, GhrR KO mice demonstrated both a significantly increased glucose infusion rate and significantly reduced insulin requirement for maintenance of the HG clamp, consistent with their relative insulin sensitivity. In HI-E clamps, both LFD-fed and HFD-fed GhrR KO mice showed higher peripheral insulin sensitivity relative to WT littermates as indicated by a significant increase in insulin-stimulated glucose disposal (Rd, and decreased hepatic glucose production (HGP. HFD-fed GhrR KO mice showed a marked increase in peripheral tissue glucose uptake in a variety of tissues, including skeletal muscle, brown adipose tissue and white adipose tissue. GhrR KO mice fed a HFD also showed a modest, but significant decrease in conversion of pyruvate to glucose, as would be anticipated if these mice displayed increased liver insulin sensitivity. Additionally, the levels of UCP2 and UCP1 were reduced in the liver and BAT, respectively, in GhrR KO mice relative to WT mice. Conclusions These results indicate that improved glucose homeostasis of GhrR KO mice is

  14. Charlie's Clamp.

    Science.gov (United States)

    Tarino, Janet Z.

    1998-01-01

    Presents a version of the crush-the-can demonstration which is a hands-on activity in which students use an inexpensive, easily made holder for the can called Charlie's clamp. Includes some suggestions for the follow-up discussion. (DDR)

  15. Euglycemic Diabetic Ketoacidosis in a Patient with Cocaine Intoxication

    Directory of Open Access Journals (Sweden)

    Asma Abu-Abed Abdin

    2016-01-01

    Full Text Available Diabetic ketoacidosis (DKA is characterized by elevated anion gap metabolic acidosis, hyperglycemia, and elevated ketones in urine and blood. Hyperglycemia is a key component of DKA; however, a subset of DKA patients can present with near-normal blood glucose, an entity described as “euglycemic DKA.” This rare phenomenon is thought to be due to starvation and food restriction in insulin dependent diabetic patients. Cocaine abuse is considered a trigger for development of DKA. Cocaine also has anorexic effects. We describe an interesting case of euglycemic DKA in a middle-aged diabetic female presenting with elevated anion gap metabolic acidosis, with near-normal blood glucose, in the settings of noncompliance to insulin and cocaine abuse. We have postulated that cocaine abuse was implicated in the pathophysiology of euglycemic DKA in this case. This case highlights complex physiological interplay between type-1 diabetes, noncompliance to insulin, and cocaine abuse leading to DKA, with starvation physiology causing development of euglycemic DKA.

  16. A cross-sectional characterization of insulin resistance by phenotype and insulin clamp in East Asian Americans with type 1 and type 2 diabetes.

    Directory of Open Access Journals (Sweden)

    William C Hsu

    Full Text Available OBJECTIVE: Classic features of type 1 and type 2 diabetes may not apply in Asian Americans, due to shared absence of common HLA DR-DQ genotype, low prevalence of positive anti-islet antibodies and low BMI in both types of diabetes. Our objective was to characterize diabetic phenotypes in Asian Americans by clamp and clinical features. MATERIALS/METHODS: This was a cross-sectional study conducted in a referral center. Thirty East young Asian American adult volunteers (27.6±5.5 years with type 1, type 2 diabetes or controls underwent hyperinsulinemic euglycemic clamp to assess insulin resistance and DEXA to assess adiposity. RESULTS: Gender, BMI, waist/hip ratio, leptin, LDL, anti-GAD, anti-IA2 antibodies and C-reactive protein were similar among three groups. Serum C-peptide, adiponectin, free fatty acid, HDL concentrations and truncal fat by DEXA, were different between diabetic groups. Glucose disposal rate by clamp was lowest in type 2 diabetes, followed by type 1 diabetes and controls (5.43±2.70, 7.62±2.59, 8.61±2.37 mg/min/kg, respectively, p = 0.001. Free fatty acid concentration universally plummeted during steady state of the clamp procedure regardless of diabetes types in all three groups. Adipocyte fatty acid binding protein in the entire cohort (r = -0.625, p = 0.04 and controls (r = -0.869, p = 0.046 correlated best with insulin resistance, independent of BMI. CONCLUSIONS: Type 2 diabetes in Asian Americans was associated with insulin resistance despite having low BMI as type 1 diabetes, suggesting a potential role for targeting insulin resistance apart from weight loss. Adipocyte fatty acid binding protein, strongly associated with insulin resistance, independent of adiposity in the young Asian American population, may potentially serve as a biomarker to identify at-risk individuals. Larger studies are needed to confirm this finding.

  17. Combined metformin-associated lactic acidosis and euglycemic ketoacidosis.

    Science.gov (United States)

    Schwetz, Verena; Eisner, Florian; Schilcher, Gernot; Eller, Kathrin; Plank, Johannes; Lind, Alice; Pieber, Thomas R; Mader, Julia K; Eller, Philipp

    2017-09-01

    In renal failure metformin can lead to lactic acidosis. Additional inhibition of hepatic gluconeogenesis by accumulation of the drug may aggravate fasting-induced ketoacidosis. We report the occurrence of metformin-associated lactic acidosis (MALA) with concurrent euglycemic ketoacidosis (MALKA) in three patients with renal failure. Patient 1: a 78-year-old woman (pH = 6.89, lactic acid 22 mmol/l, serum ketoacids 7.4 mmol/l and blood glucose 63 mg/dl) on metformin and insulin treatment. Patient 2: a 79-year-old woman on metformin treatment (pH = 6.80, lactic acid 14.7 mmol/l, serum ketoacids 6.4 mmol/l and blood glucose 76 mg/dl). Patient 3: a 71-year-old man on metformin, canagliflozin and liraglutide treatment (pH = 7.21, lactic acid 5.9 mmol/l, serum ketoacids 16 mmol/l and blood glucose 150 mg/dl). In all patients, ketoacidosis receded on glucose infusion and renal replacement therapy. This case series highlights the parallel occurrence of MALA and euglycemic ketoacidosis, the latter exceeding ketosis due to starvation, suggesting a metformin-triggered inhibition of gluconeogenesis. Affected patients benefit from glucose infusion counteracting suppressed hepatic gluconeogenesis.

  18. Clamp usable as jig and lifting clamp

    Science.gov (United States)

    Tsuyama, Yoshizo

    1976-01-01

    There is provided a clamp which is well suited for use as a lifting clamp for lifting and moving materials of assembly in a shipyard, etc. and as a pulling jig in welding and other operations. The clamp comprises a clamp body including a shackle for engagement with a pulling device and a slot for receiving an article, and a pair of jaws provided on the leg portions of the clamp body on the opposite sides of the slot to grip the article in the slot, one of said jaws consisting of a screw rod and the other jaw consisting of a swivel jaw with a spherical surface, whereby when the article clamped in the slot by the pair of jaws tends to slide in any direction with respect to the clamp body, the article is more positively gripped by the pair of jaws.

  19. Insulin resistance in obesity can be reliably identified from fasting plasma insulin

    NARCIS (Netherlands)

    ter Horst, K. W.; Gilijamse, P. W.; Koopman, K. E.; de Weijer, B. A.; Brands, M.; Kootte, R. S.; Romijn, J. A.; Ackermans, M. T.; Nieuwdorp, M.; Soeters, M. R.; Serlie, M. J.

    2015-01-01

    Insulin resistance is the major contributor to cardiometabolic complications of obesity. We aimed to (1) establish cutoff points for insulin resistance from euglycemic hyperinsulinemic clamps (EHCs), (2) identify insulin-resistant obese subjects and (3) predict insulin resistance from routinely

  20. Genetic variants associated with glycine metabolism and their role in insulin sensitivity and type 2 diabetes

    DEFF Research Database (Denmark)

    Xie, Weijia; Wood, Andrew R; Lyssenko, Valeriya

    2013-01-01

    Circulating metabolites associated with insulin sensitivity may represent useful biomarkers, but their causal role in insulin sensitivity and diabetes is less certain. We previously identified novel metabolites correlated with insulin sensitivity measured by the hyperinsulinemic-euglycemic clamp....

  1. Diabetic euglycemic ketoacidosis in newly diagnosed type 1 diabetes mellitus during Ramadan fasting.

    Science.gov (United States)

    Baş, Veysel Nijat; Uytun, Salih; Torun, Yasemin Altuner

    2015-03-01

    Real euglycemic diabetic ketoacidosis [DKA; blood glucose importance of evaluation of acid-base state, urine glucose, and ketone values at the application in all newly diagnosed type 1 diabetic patients with normal glucose levels by defining euglycemic DKA that resulted from long-lasting starvation during Ramadan fasting in a newly diagnosed 14-year-old male patient.

  2. Quick-attach clamp

    Science.gov (United States)

    Vano, A. E.

    1968-01-01

    Clamp of the slideable jaw type can be applied to moving lines such as cables or ropes. The clamp has a trigger-operated jaw that can be attached to a redrop parachute on a moving tow cable. The trigger mechanism maintains the jaws retracted in the housing until they are released for clamping.

  3. Association between irisin and homocysteine in euglycemic and diabetic subjects.

    Science.gov (United States)

    Alis, Rafael; Sanchis-Gomar, Fabian; Pareja-Galeano, Helios; Hernández-Mijares, Antonio; Romagnoli, Marco; Víctor, Víctor M; Rocha, Milagros

    2014-12-01

    The aim of study was to explore whether a relationship exists between homocysteine and irisin in type 2 diabetes (T2D) patients-a population with a high risk of developing cardiovascular disease-and euglycemic controls. 69 T2D patients and 75 control subjects (adjusted by body mass index (BMI)) were included in the study. Irisin and homocysteine concentrations and anthropometric and biochemical variables were determined. Levels of homocysteine were significantly higher (11.0±3.0 vs 12.4±4.2 μmol/l) and levels of irisin were lower (279±58 vs 263±38 ng/ml) in T2D patients. When both T2D and controls were considered, irisin was found to correlate only with homocysteine (r=-0.215; p=0.011). Moreover, a decreasing trend in irisin levels was observed according to homocysteine tertile (p=0.034). Our results provide evidence of an association between irisin and homocysteine, which may be due to nicotinamide metabolism. The clinical significance of this relationship is unclear, but our findings may prompt further mechanistic research to investigate the role played by irisin in vascular disorders. Copyright © 2014 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  4. Photovoltaic panel clamp

    Science.gov (United States)

    Brown, Malcolm P.; Mittan, Margaret Birmingham; Miros, Robert H. J.; Stancel, Robert

    2013-03-19

    A photovoltaic panel clamp includes an upper and lower section. The interface between the assembled clamp halves and the module edge is filled by a flexible gasket material, such as EPDM rubber. The gasket preferably has small, finger like protrusions that allow for easy insertion onto the module edge while being reversed makes it more difficult to remove them from the module once installed. The clamp includes mounting posts or an integral axle to engage a bracket. The clamp also may include a locking tongue to secure the clamp to a bracket.

  5. Radial wedge flange clamp

    Science.gov (United States)

    Smith, Karl H.

    2002-01-01

    A radial wedge flange clamp comprising a pair of flanges each comprising a plurality of peripheral flat wedge facets having flat wedge surfaces and opposed and mating flat surfaces attached to or otherwise engaged with two elements to be joined and including a series of generally U-shaped wedge clamps each having flat wedge interior surfaces and engaging one pair of said peripheral flat wedge facets. Each of said generally U-shaped wedge clamps has in its opposing extremities apertures for the tangential insertion of bolts to apply uniform radial force to said wedge clamps when assembled about said wedge segments.

  6. Clamp Restrains Pressure Line

    Science.gov (United States)

    Aliberti, J. A.

    1982-01-01

    Safety restraint protects people and property if a high-pressure fitting fails. As long as pressure line remains attached at the fitting, clamp exerts essentially no force on hose. If fitting fails, force of fluid leaving free end of hose causes the cam on the clamp to compress hose with a positive locking action.

  7. Quick action clamp

    Science.gov (United States)

    Calco, Frank S. (Inventor)

    1991-01-01

    A quick release toggle clamp that utilizes a spring that requires a deliberate positive action for disengagement is presented. The clamp has a sliding bolt that provides a latching mechanism. The bolt is moved by a handle that tends to remain in an engaged position while under tension.

  8. Clamp-mount device

    Science.gov (United States)

    Clark, K. H. (Inventor)

    1983-01-01

    A clamp-mount device is disclosed for mounting equipment to an associated I-beam and the like structural member of the type having oppositely extending flanges wherein the device comprises a base and a pair of oppositely facing clamping members carried diagonally on the base clamping flanges therebetween and having flange receiving openings facing one another. Lock means are carried diagonally by the base opposite the clamping members locking the flanges in the clamping members. A resilient hub is carried centrally of the base engaging and biasing a back side of the flanges maintaining tightly clamped and facilitating use on vertical as well as horizontal members. The base turns about the hub to receive the flanges within the clamping members. Equipment may be secured to the base by any suitable means such as bolts in openings. Slidable gate latches secure the hinged locks in an upright locking position. The resilient hub includes a recess opening formed in the base and a rubber-like pad carried in this opening being depressably and rotatably carried therein.

  9. Bend-absorbing clamp

    Science.gov (United States)

    Abbott, J. R.; Valencia, B., Jr.

    1979-01-01

    Compact, inexpensive clamp for flexible cables or rigid tubes absorbs vibrations and other motion. It accomodates wide range of dimensions, and saves space by eliminating pigtails or bellows commonly used to absorb linear movement or vibrations

  10. Persistent hyperinsulinemic hypoglycemia of infancy: An overview of current concepts

    Directory of Open Access Journals (Sweden)

    Prabudh Goel

    2012-01-01

    Full Text Available Persistent hyperinsulinemic hypoglycemia of infancy (PHHI is relatively rare but one of the most important causes of severe neonatal hypoglycemia. Recognition of this entity becomes important due to the fact that the hypoglycemia is so severe and frequent that it may lead to severe neurological damage in the infant manifesting as mental or psychomotor retardation or even a life-threatening event if not recognized and treated effectively in time. Near-total pancreatectomy may be required for patients with intractable hypoglycemia despite medical treatment; however, that may result in diabetes mellitus or recurrent postoperative hypoglycemia. This review aims to consolidate the traditional concepts and current information related to the pathogenesis and management of PHHI.

  11. How a DNA polymerase clamp loader opens a sliding clamp

    OpenAIRE

    Kelch, Brian A.; Makino, Debora L.; O’Donnell, Mike; Kuriyan, John

    2011-01-01

    Processive chromosomal replication relies on sliding DNA clamps, which are loaded onto DNA by pentameric clamp loader complexes belonging to the AAA+ family of ATPases. We present structures for the ATP-bound state of the clamp loader complex from bacteriophage T4, bound to an open clamp and primer-template DNA. The clamp loader traps a spiral conformation of the open clamp so that both the loader and the clamp match the helical symmetry of DNA. One structure reveals that ATP has been hydroly...

  12. How a DNA polymerase clamp loader opens a sliding clamp

    Science.gov (United States)

    Kelch, Brian A.; Makino, Debora L.; O’Donnell, Mike; Kuriyan, John

    2012-01-01

    Processive chromosomal replication relies on sliding DNA clamps, which are loaded onto DNA by pentameric clamp loader complexes belonging to the AAA+ family of ATPases. We present structures for the ATP-bound state of the clamp loader complex from bacteriophage T4, bound to an open clamp and primer-template DNA. The clamp loader traps a spiral conformation of the open clamp so that both the loader and the clamp match the helical symmetry of DNA. One structure reveals that ATP has been hydrolyzed in one subunit, and suggests that clamp closure and ejection of the loader involves disruption of the ATP-dependent match in symmetry. The structures explain how synergy between the loader, the clamp and DNA can trigger ATP hydrolysis and release of the closed clamp on DNA. PMID:22194570

  13. Clamp Secures Material to Cylinder Base

    Science.gov (United States)

    Tam, Y. S.

    1985-01-01

    Two-part clamp assembled around pipe or post, ends not free. Resistance welding clamp to post and locking ring to lower part of clamp permanently fixes clamp in place, preventing thermal cycling from loosening clamp.

  14. Clinical Features and Causes of Endogenous Hyperinsulinemic Hypoglycemia in Korea

    Directory of Open Access Journals (Sweden)

    Chang-Yun Woo

    2015-04-01

    Full Text Available BackgroundEndogenous hyperinsulinemic hypoglycemia (EHH is characterized by an inappropriately high plasma insulin level, despite a low plasma glucose level. Most of the EHH cases are caused by insulinoma, whereas nesidioblastosis and insulin autoimmune syndrome (IAS are relatively rare.MethodsTo evaluate the relative frequencies of various causes of EHH in Korea, we retrospectively analyzed 84 patients who were diagnosed with EHH from 1998 to 2012 in a university hospital.ResultsAmong the 84 EHH patients, 74 patients (88%, five (6%, and five (6% were diagnosed with insulinoma, nesidioblastosis or IAS, respectively. The most common clinical manifestation of EHH was neuroglycopenic symptoms. Symptom duration before diagnosis was 14.5 months (range, 1 to 120 months for insulinoma, 1.0 months (range, 6 days to 7 months for nesidioblastosis, and 2.0 months (range, 1 to 12 months for IAS. One patient, who was diagnosed with nesidioblastosis in 2006, underwent distal pancreatectomy but was later determined to be positive for insulin autoantibodies. Except for one patient who was diagnosed in 2007, the remaining three patients with nesidioblastosis demonstrated severe hyperinsulinemia (157 to 2,719 µIU/mL, which suggests that these patients might have had IAS, rather than nesidioblastosis.ConclusionThe results of this study suggest that the prevalence of IAS may be higher in Korea than previously thought. Therefore, measurement of insulin autoantibody levels is warranted for EHH patients, especially in patients with very high plasma insulin levels.

  15. Clamp for detonating fuze

    Science.gov (United States)

    Holderman, E. J.

    1968-01-01

    Quick acting clamp provides physical support for a closely confined detonating fuse in an application requiring removal and replacement at frequent intervals during test. It can be designed with a base of any required strength and configuration to permit the insertion of an object.

  16. Mechanism of opening a sliding clamp.

    Science.gov (United States)

    Douma, Lauren G; Yu, Kevin K; England, Jennifer K; Levitus, Marcia; Bloom, Linda B

    2017-09-29

    Clamp loaders load ring-shaped sliding clamps onto DNA where the clamps serve as processivity factors for DNA polymerases. In the first stage of clamp loading, clamp loaders bind and stabilize clamps in an open conformation, and in the second stage, clamp loaders place the open clamps around DNA so that the clamps encircle DNA. Here, the mechanism of the initial clamp opening stage is investigated. Mutations were introduced into the Escherichia coli β-sliding clamp that destabilize the dimer interface to determine whether the formation of an open clamp loader-clamp complex is dependent on spontaneous clamp opening events. In other work, we showed that mutation of a positively charged Arg residue at the β-dimer interface and high NaCl concentrations destabilize the clamp, but neither facilitates the formation of an open clamp loader-clamp complex in experiments presented here. Clamp opening reactions could be fit to a minimal three-step 'bind-open-lock' model in which the clamp loader binds a closed clamp, the clamp opens, and subsequent conformational rearrangements 'lock' the clamp loader-clamp complex in a stable open conformation. Our results support a model in which the E. coli clamp loader actively opens the β-sliding clamp. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  17. Tumor necrosis factor alpha is associated with insulin-mediated suppression of free fatty acids and net lipid oxidation in HIV-infected patients with lipodystrophy

    DEFF Research Database (Denmark)

    Haugaard, Steen B; Andersen, Ove; Pedersen, SB

    2006-01-01

    with lipodystrophy (LIPO) and those without (controls). LIPOX was estimated by indirect calorimetry during fasting and steady state of a hyperinsulinemic euglycemic clamp in 36 (18 LIPO and 18 controls) normoglycemic HIV-infected men on highly active antiretroviral therapy. In LIPO, TNF-alpha correlated with clamp...

  18. Hand-Held Power Clamp

    Science.gov (United States)

    Clancy, J. P.

    1985-01-01

    Tool furnishes large pushing or pulling forces. Device includes two clamping blocks, two clamping plates, and a motor-driven linear actuator with selflocking screw shaft. Power clamp exerts opening or closing force at push of switch. Tool approximately 1 m long. Originally designed to secure payload aboard Space Shuttle, operated with one hand to apply opening or closing force of up to 1,000 lb (4,400 N). Clamp has potential applications as end effector for industrial robots and in rescue work to push or pull wreckage with great force.

  19. The Escherichia coli Clamp Loader Can Actively Pry Open the β-Sliding Clamp*

    Science.gov (United States)

    Paschall, Christopher O.; Thompson, Jennifer A.; Marzahn, Melissa R.; Chiraniya, Ankita; Hayner, Jaclyn N.; O'Donnell, Mike; Robbins, Arthur H.; McKenna, Robert; Bloom, Linda B.

    2011-01-01

    Clamp loaders load ring-shaped sliding clamps onto DNA. Once loaded onto DNA, sliding clamps bind to DNA polymerases to increase the processivity of DNA synthesis. To load clamps onto DNA, an open clamp loader-clamp complex must form. An unresolved question is whether clamp loaders capture clamps that have transiently opened or whether clamp loaders bind closed clamps and actively open clamps. A simple fluorescence-based clamp opening assay was developed to address this question and to determine how ATP binding contributes to clamp opening. A direct comparison of real time binding and opening reactions revealed that the Escherichia coli γ complex binds β first and then opens the clamp. Mutation of conserved “arginine fingers” in the γ complex that interact with bound ATP decreased clamp opening activity showing that arginine fingers make an important contribution to the ATP-induced conformational changes that allow the clamp loader to pry open the clamp. PMID:21971175

  20. Fasting insulin sensitivity indices are not better than routine clinical variables at predicting insulin sensitivity among Black Africans: a clamp study in sub-Saharan Africans

    Science.gov (United States)

    2014-01-01

    Background We aimed to evaluate the predictive utility of common fasting insulin sensitivity indices, and non-laboratory surrogates [BMI, waist circumference (WC) and waist-to-height ratio (WHtR)] in sub-Saharan Africans without diabetes. Methods We measured fasting glucose and insulin, and glucose uptake during 80/mU/m2/min euglycemic clamp in 87 Cameroonians (51 men) aged (SD) 34.6 (11.4) years. We derived insulin sensitivity indices including HOMA-IR, quantitative insulin sensitivity check index (QUICKI), fasting insulin resistance index (FIRI) and glucose-to-insulin ratio (GIR). Indices and clinical predictors were compared to clamp using correlation tests, robust linear regressions and agreement of classification by sex-specific thirds. Results The mean insulin sensitivity was M = 10.5 ± 3.2 mg/kg/min. Classification across thirds of insulin sensitivity by clamp matched with non-laboratory surrogates in 30-48% of participants, and with fasting indices in 27-51%, with kappa statistics ranging from −0.10 to 0.26. Fasting indices correlated significantly with clamp (/r/=0.23-0.30), with GIR performing less well than fasting insulin and HOMA-IR (both p insulin sensitivity indices are modest predictors of insulin sensitivity measured by euglycemic clamp, and do not perform better than clinical surrogates in this population. PMID:25106496

  1. Octreotide therapy and restricted fetal growth: pregnancy in familial hyperinsulinemic hypoglycemia

    Directory of Open Access Journals (Sweden)

    Marianne Geilswijk

    2017-02-01

    Full Text Available Hypoglycemia during pregnancy can have serious health implications for both mother and fetus. Although not generally recommended in pregnancy, synthetic somatostatin analogues are used for the management of blood glucose levels in expectant hyperinsulinemic mothers. Recent reports suggest that octreotide treatment in pregnancy, as well as hypoglycemia in itself, may pose a risk of fetal growth restriction. During pregnancy, management of blood glucose levels in familial hyperinsulinemic hypoglycemia thus forms a medical dilemma. We report on pregnancy outcomes in a woman with symptomatic familial hyperinsulinemic hypoglycemia, type 3. During the patient’s first pregnancy with a viable fetus octreotide treatment was instituted in gestational age 23 weeks to prevent severe hypoglycemic incidences. Fetal growth velocity declined, and at 37 weeks of gestation, intrauterine growth retardation was evident. During the second pregnancy with a viable fetus, blood glucose levels were managed through dietary intervention alone. Thus, the patient was advised to take small but frequent meals high in fiber and low in carbohydrates. Throughout pregnancy, no incidences of severe hypoglycemia occurred and fetal growth velocity was normal. We conclude that octreotide treatment during pregnancy may pose a risk of fetal growth restriction and warrants careful consideration. In some cases of familial hyperinsulinemic hypoglycemia, blood glucose levels can be successfully managed through diet only, also during pregnancy.

  2. [Feasibility of domestic (18)F-DOPA PET/CT scanning in the differential diagnosis of pancreatic lesions in children with hyperinsulinemic hypoglycemia].

    Science.gov (United States)

    Zhang, M Y; Ge, J J; Pei, Z; Dong, K R; Chen, L; Wang, X; Zhang, Z W; Li, X J; Xi, L; Cheng, R Q; Lu, G P; Guan, Y H; Luo, F H

    2017-10-02

    Objective: To study the feasibility of (18)F-fluoro-L-dihydroxyphenylalanine positron emission tomography/Computed tomography ((18)F-DOPA PET/CT) scanning in the localization and differential diagnosing of focal versus diffuse form of pancreas lesions in patients with hyperinsulinemic hypoglycemia (HH). Method: Twenty-four patients were diagnosed with HH between January, 2016 and February, 2017 in the Department of Pediatric Endocrinology and Inherited Metabolic Diseases, Children's Hospital of Fudan University using an integrated clinical and biochemical diagnostic protocol, domestic (18)F-DOPA PET/CT imaging technique were applied after MRI and ultrasound failed to detect pancreas lesions. Pancreas (18)F-DOPA standardized uptake values (SUV) were measured, and pancreas' lesions were dually analyzed via visual method and pancreas percentage SUV method. Among these patients, 9 patients received surgical pancreatic lesion resections, the correlations among surgical outcomes, histopathological findings and (18)F-DOPA PET/CT scan results were analyzed. Result: Seven patients were detected with focal form of pancreas lesions, the mean peak of SUV was 4.7±1.7(2.6-7.1), and 17 patients were found to have diffuse form lesions after (18)F-DOPA-PET/CT scanning. Among the 24 cases, 9 patients (7 showed focal and 2 showed diffuse (18)F-DOPA PET/CT pancreatic uptake)were euglycemic without any medical support after surgery; the resected pancreatic tissue histopathological results were consistent with that of PET/CT imaging. Only one patient, who responded to medical treatment before surgery, had temporary hyperglycemia after operation. Conclusion: Domestic (18)F-DOPA PET/CT could successfully locate and differentiate the pancreatic lesions and thus improve the success of surgery.

  3. Internal V-Band Clamp

    Science.gov (United States)

    Vaughn, Mark R.; Hafenrichter, Everett S.; Chapa, Agapito C.; Harris, Steven M.; Martinez, Marcus J.; Baty, Roy S.

    2006-02-28

    A system for clamping two tubular members together in an end-to-end relationship uses a split ring with a V-shaped outer rim that can engage a clamping surface on each member. The split ring has a relaxed closed state where the ends of the ring are adjacent and the outside diameter of the split ring is less than the minimum inside diameter of the members at their ends. The members are clamped when the split ring is spread into an elastically stretched position where the ring rim is pressed tightly against the interior surfaces of the members. Mechanisms are provided for removing the spreader so the split ring will return to the relaxed state, releasing the clamped members.

  4. Micromachined patch-clamp apparatus

    Science.gov (United States)

    Okandan, Murat

    2012-12-04

    A micromachined patch-clamp apparatus is disclosed for holding one or more cells and providing electrical, chemical, or mechanical stimulation to the cells during analysis with the patch-clamp technique for studying ion channels in cell membranes. The apparatus formed on a silicon substrate utilizes a lower chamber formed from silicon nitride using surface micromachining and an upper chamber formed from a molded polymer material. An opening in a common wall between the chambers is used to trap and hold a cell for analysis using the patch-clamp technique with sensing electrodes on each side of the cell. Some embodiments of the present invention utilize one or more electrostatic actuators formed on the substrate to provide mechanical stimulation to the cell being analyzed, or to provide information about mechanical movement of the cell in response to electrical or chemical stimulation.

  5. How to Assess the Quality of Glucose Clamps? Evaluation of Clamps Performed With ClampArt, a Novel Automated Clamp Device.

    Science.gov (United States)

    Benesch, Carsten; Heise, Tim; Klein, Oliver; Heinemann, Lutz; Arnolds, Sabine

    2015-07-01

    There are no widely accepted parameters to assess the quality of glucose clamps. Thus, we selected different parameters describing clamp quality. These parameters were then evaluated in glucose clamps carried out with ClampArt, a novel CE-marked, state-of-the-art fully automated glucose clamp device employing continuous blood glucose (BG) measurements and minute-by-minute adaptations of glucose infusion rate (GIR). Thirty-nine glucose clamps were performed in 10 healthy and 29 subjects with type 1 diabetes (T1DM) (total duration 583 h). ClampArt-based BG measurements were compared with those obtained with a laboratory reference method. Clamp quality was assessed by 5 parameters: (1) difference (mg/dl) of all paired BG measurements of ClampArt versus reference method ("trueness"), (2) coefficient of variation (CV, %) of ClampArt's BG measurements at target clamp level ("precision"), (3) mean absolute relative difference (MARD, %) at target clamp level ("accuracy"), (4) difference (mg/dl) between ClampArt and target BG ("control deviation"), and (5) percentage operational time ("utility"). ClampArt-based BG measurements showed a trueness of 1.2 ± 2.5 mg/dl. CV and MARD at target BG were 5.5 ± 2.1% and 5.3 ± 2.3%, respectively. There were only small deviations from target level (1.2 ± 1.6 mg/dl). Operational time was as high as 95.4% ± 4.1% (means ± SD). The selected parameters seem to be adequate to characterize clamp quality. The novel, fully automated clamp device ClampArt achieves high clamp quality, which in future trials should be compared with other (automated and manual) clamp methods. © 2015 Diabetes Technology Society.

  6. Intimal injury from arterial clamps.

    Science.gov (United States)

    Slayback, J B; Bowen, W W; Hinshaw, D B

    1976-08-01

    Preliminary experimental data have been presented indicating that intimal injury of some degree is a virtually constant finding at the site of application of any arterial occluding clamp. These injuries vary from intimal distortion to complete fracture into the media of the vessel. The degree of injury appears directly proportional to the amount of pressure exerted through a given clamp. Atherosclerotic arteries are particularly subject to severe degrees of intimal injury. Preliminary observations suggest that heparin is not helpful in preventing platelet aggregation and initial thrombus formation at intimal injury sites. The problem of anticoagulation at such injury sites is being studied further.

  7. The association between TNF-α and insulin resistance in euglycemic women.

    LENUS (Irish Health Repository)

    Walsh, Jennifer M

    2013-10-01

    Chronic low levels of inflammation have links to obesity, diabetes and insulin resistance. We sought to assess the relationship between cytokine tumor necrosis factor (TNF-α) and insulin resistance in a healthy, euglycemic population. This is a prospective study of 574 non-diabetic mother and infant pairs. Maternal body mass index (BMI), TNF-α, glucose and insulin were measured in early pregnancy and at 28 weeks. Insulin resistance was calculated by HOMA index. At delivery birthweight was recorded and cord blood analysed for fetal C-peptide and TNF-α. In a multivariate model, maternal TNF-α in early pregnancy was predicted by maternal insulin resistance at the same time-point, (β=0.54, p<0.01), and maternal TNF-α at 28 weeks was predicted by maternal insulin resistance in early pregnancy (β=0.24, p<0.01) and at 28 weeks (β=0.39, p<0.01). These results, in a large cohort of healthy, non-diabetic women have shown that insulin resistance, even at levels below those diagnostic of gestational diabetes, is associated with maternal and fetal inflammatory response. These findings have important implications for defining the pathways of fetal programming of later metabolic syndrome and childhood obesity.

  8. Fasting insulin sensitivity indices are not better than routine clinical variables at predicting insulin sensitivity among Black Africans: a clamp study in sub-Saharan Africans.

    Science.gov (United States)

    Sobngwi, Eugene; Kengne, Andre-Pascal; Echouffo-Tcheugui, Justin B; Choukem, Simeon; Sobngwi-Tambekou, Joelle; Balti, Eric V; Pearce, Mark S; Siaha, Valentin; Mamdjokam, Aissa S; Effoe, Valery; Lontchi-Yimagou, Eric; Donfack, Oliver T; Atogho-Tiedeu, Barbara; Boudou, Philippe; Gautier, Jean-Francois; Mbanya, Jean-Claude

    2014-08-09

    We aimed to evaluate the predictive utility of common fasting insulin sensitivity indices, and non-laboratory surrogates [BMI, waist circumference (WC) and waist-to-height ratio (WHtR)] in sub-Saharan Africans without diabetes. We measured fasting glucose and insulin, and glucose uptake during 80/mU/m2/min euglycemic clamp in 87 Cameroonians (51 men) aged (SD) 34.6 (11.4) years. We derived insulin sensitivity indices including HOMA-IR, quantitative insulin sensitivity check index (QUICKI), fasting insulin resistance index (FIRI) and glucose-to-insulin ratio (GIR). Indices and clinical predictors were compared to clamp using correlation tests, robust linear regressions and agreement of classification by sex-specific thirds. The mean insulin sensitivity was M = 10.5 ± 3.2 mg/kg/min. Classification across thirds of insulin sensitivity by clamp matched with non-laboratory surrogates in 30-48% of participants, and with fasting indices in 27-51%, with kappa statistics ranging from -0.10 to 0.26. Fasting indices correlated significantly with clamp (/r/=0.23-0.30), with GIR performing less well than fasting insulin and HOMA-IR (both p fasting indices (/r/=0.38-0.43). Combinations of fasting indices and clinical predictors explained 25-27% of variation in clamp values. Fasting insulin sensitivity indices are modest predictors of insulin sensitivity measured by euglycemic clamp, and do not perform better than clinical surrogates in this population.

  9. Insulin resistance in SD rats chronically treated with ethanol

    Science.gov (United States)

    We have previously demonstrated that hepatic insulin signaling is disrupted in Sprague-Dawley (SD) rats fed EtOH-containing diets by total enteral nutrition (TEN). To determine if whole body insulin resistance could be demonstrated in the TEN model, we conducted euglycemic-hyperinsulinemic clamp st...

  10. Mathematical modeling of the glucose-insulin system

    DEFF Research Database (Denmark)

    Palumbo, Pasquale; Ditlevsen, Susanne; Bertuzzi, Alessandro

    2013-01-01

    of pancreatic insulin production, with a oarser/finer level of detail ranging over cellular and subcellular scales, to short-term organ/tissue models accounting for the intra-venous and the oral glucose tolerance tests as well as for the euglycemic hyperinsulinemic clamp, to total-body, long-term diabetes...

  11. Exercise increases human skeletal muscle insulin sensitivity via coordinated increases in microvascular perfusion and molecular signaling

    DEFF Research Database (Denmark)

    Sjøberg, Kim Anker; Frøsig, Christian; Kjøbsted, Rasmus

    2017-01-01

    Insulin resistance is a major health risk and although exercise clearly improves skeletal muscle insulin sensitivity, the mechanisms are unclear. Here we show that initiation of a euglycemic hyperinsulinemic clamp four hours after single-legged exercise in humans increased microvascular perfusion...

  12. Protection from obesity and insulin resistance in mice overexpressing human apolipoprotein C1

    NARCIS (Netherlands)

    Jong, M. C.; Voshol, P. J.; Muurling, M.; Dahlmans, V. E.; Romijn, J. A.; Pijl, H.; Havekes, L. M.

    2001-01-01

    Apolipoprotein (APO) C1 is a 6.6-kDa protein present in plasma and associated with lipoproteins. Using hyperinsulinemic-euglycemic clamp tests, we previously found that in APOC1 transgenic mice, the whole-body insulin-mediated glucose uptake is increased concomitant with a decreased fatty acid

  13. Combined intervention with pioglitazone and n-3 fatty acids in metformin-treated type 2 diabetic patients: improvement of lipid metabolism

    Czech Academy of Sciences Publication Activity Database

    Veleba, J.; Kopecký Jr., J.; Janovská, Petra; Kuda, Ondřej; Horáková, Olga; Malínská, H.; Kazdová, L.; Oliyarnyk, O.; Škop, V.; Trnovská, J.; Hájek, M.; Škoch, A.; Flachs, Pavel; Bardová, Kristina; Rossmeisl, Martin; Olza, J.; de Castro, S. G.; Calder, P. C.; Gardlo, Alžběta; Fišerová, E.; Jensen, J.; Bryhn, M.; Kopecký, Jan; Pelikánová, T.

    2015-01-01

    Roč. 12, Dec 2 (2015), s. 52 ISSN 1743-7075 R&D Projects: GA MZd(CZ) NT13763 Institutional support: RVO:67985823 Keywords : eicosapentaenoic acid * docosahexaenoic acid * indirect calorimetry * meal test * humans * hyperinsulinemic-euglycemic clamp Subject RIV: FB - Endocrinology, Diabetology, Metabolism, Nutrition Impact factor: 3.280, year: 2015

  14. Effect of antioxidant supplementation on insulin sensitivity in response to endurance exercise training

    DEFF Research Database (Denmark)

    Yfanti, Christina; Nielsen, Anders R; Åkerström, Thorbjörn

    2011-01-01

    ) group that both underwent a supervised intense endurance-training program, 5 times per week for 12 weeks. A 3-hour euglycemic-hyperinsulinemic clamp, a maximal oxygen consumption (VO(2max)) and maximal power output (P(max)) test, and body composition measurements (fat mass, fat-free mass) were performed...

  15. Effect of training on insulin sensitivity of glucose uptake and lipolysis in human adipose tissue

    DEFF Research Database (Denmark)

    Stallknecht, Bente; Larsen, J J; Mikines, K J

    2000-01-01

    Training increases insulin sensitivity of both whole body and muscle in humans. To investigate whether training also increases insulin sensitivity of adipose tissue, we performed a three-step hyperinsulinemic, euglycemic clamp in eight endurance-trained (T) and eight sedentary (S) young men [insu...

  16. Co-ordination of hepatic and adipose tissue lipid metabolism after oral glucose

    DEFF Research Database (Denmark)

    Bülow, J; Simonsen, L; Wiggins, D

    1999-01-01

    The integration of lipid metabolism in the splanchnic bed and in subcutaneous adipose tissue before and after ingestion of a 75 g glucose load was studied by Fick's principle in seven healthy subjects. Six additional subjects were studied during a hyperinsulinemic euglycemic clamp. Release of non...

  17. Clamp wins pipe repair prize

    Energy Technology Data Exchange (ETDEWEB)

    Anon.

    2001-04-01

    This paper describes the permanent pipeline repair system, developed by Tekmar, which is powered by seawater hydraulics and is easily installed and tested by any workclass remotely operated vehicle (rov). Details are given of the two main components of the system, namely, the diverless high pressure split repair clamp and the rov-operated tool to install it.

  18. [Cardiac performance during insulin clamp: its evaluation by bioimpedance measurement].

    Science.gov (United States)

    Ferraro, S; Bellarosa, I; Genovese, S; Desiderio, A; D'Alto, M; Trimigliozzi, P; Chiariello, M

    1997-02-01

    Ten patients with dilated cardiomyopathy, 7 ischemic heart disease and 3 idiopathic (7 males and 3 females, mean age 54.8 +/- 11 years), were studied. An insulin euglycemic clamp was performed to evaluate the influence of insulin infusion on cardiac performance. All the subjects were in NYHA functional class II-III and had an ejection fraction < 60% at a previous echocardiographic examination. They were submitted to a noninvasive continuous cardiac output monitoring by variations of thoracic impedance at three stops: a basal evaluation (T0); 1.2 mU/kg/min insulin infusion (T1); 5 mU/kg/min insulin infusion (T2). Moreover at the same times Na+, K+, hematocrit (Hct) and catecholamines were measured in the plasma. The total time required for the study was about 4 hours. Cardiac performance improved as shown by a significant increase in T1 and T2 with respect to the basal condition (T0), of ejection fraction: 57.87 +/- 11.3% (T1) vs 52.25 +/- 14.1% (T0; p < 0.01); 59.5 +/- 10.7% (T2) vs 52.25 +/- 14.1% (T0; p < 0.005); stroke volume: 2.61 +/- 0.5 l/min/m2 (T1) vs 2.01 +/- 0.75 l/min/m2 (T0; p < 0.001); 3.35 +/- 0.66 l/min/m2 (T2) vs 2.01 +/- 0.75 l/min/m2 (T0; p < 0.001) and cardiac output: 41.14 +/- 9.1 ml/m2 (T1) vs 38.83 +/- 7.1 ml/m2 (T0; p < 0.01); 46.45 +/- 8.56 ml/m2 (T2) vs 38.83 +/- 7.1 ml/m2 (T0; p < 0.001). This improvement in cardiac function was not due to an increase in preload because end diastolic volume, Hct, Na+ did not differ through the study. Similarly, afterload measured as mean blood pressure did not decrease. Cardiac function improvement was obtained in the presence of increased catecholamines. In conclusion, exogenous insulin infusion was able, in the presence of euglycemic condition, to improve cardiac performance. This improvement could be due to a direct inotropic effect of insulin perhaps mediated by its metabolic properties on cardiac muscular cells and/or by increasing the cellular intake of Ca+2.

  19. To clamp or not to clamp? Long‐term functional outcomes for elective off‐clamp laparoscopic partial nephrectomy

    National Research Council Canada - National Science Library

    Shah, Paras H; George, Arvin K; Moreira, Daniel M; Alom, Manaf; Okhunov, Zhamshid; Salami, Simpa; Waingankar, Nikhil; Schwartz, Michael J; Vira, Manish A; Richstone, Lee; Kavoussi, Louis R

    2016-01-01

    ... is to avoid ischaemia altogether by performing surgery entirely off‐clamp. Several institutions have reported superior renal functional preservation with the off‐clamp compared with the on‐clamp method after elective laparoscopic PN (LPN), where there is a solitary tumour and normal contralateral kidney and preoperative serum creatinine ...

  20. Multifrequency Excitation of a Clamped-Clamped Microbeam

    KAUST Repository

    Jaber, Nizar R.

    2016-01-20

    We present analytical and experimental investigation of an electrically actuated clamped-clamped microbeam under a twosource harmonic excitation. The first frequency is swept around the first mode of vibration where the second one is fixed. These microbeams are fabricated using polyimide as structural layer coated with nickel form top and chromium and gold layers from bottom. We demonstrate the excitation of additive and subtractive type resonance. We show that by properly tuning the frequency and the amplitude of the excitation force, the frequency bandwidth of the resonator is increased. Theoretically, we solved the eigenvalue problem for different axial forces to find the natural frequencies ratio that match the experimental values. Using Galerkin method, a reduced order model is derived to simulate the static and dynamic response of the device where using three symmetric mode shapes provided a good agreement with experimental data. © Copyright 2015 by ASME.

  1. Analysis list: Clamp [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Clamp Cell line + dm3 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/target/Clamp.1....tsv http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/target/Clamp.5.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/target/Clam...p.10.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/colo/Clamp.Cell_line.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/colo/Cell_line.gml ...

  2. Box Lesion with Single Radiofrequency Clamp.

    Science.gov (United States)

    Shaikhrezai, Kasra; Spadaccio, Cristiano; Hunter, Steven

    2017-04-12

    We introduce a maneuver to perform the box lesion as a part of Cox maze IV procedure with single radiofrequency clamp. The maneuver entails engaging the right pulmonary veins with the clamp, then advancing the clamp jaws underneath the superior vena cava through the transverse sinus to include the roof and the left pulmonary veins in the clamp. We regularly use this technique for box lesion with desirable transmurality. Georg Thieme Verlag KG Stuttgart · New York.

  3. 21 CFR 870.4450 - Vascular clamp.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Vascular clamp. 870.4450 Section 870.4450 Food and... CARDIOVASCULAR DEVICES Cardiovascular Surgical Devices § 870.4450 Vascular clamp. (a) Identification. A vascular clamp is a surgical instrument used to occlude a blood vessel temporarily. (b) Classification. Class II...

  4. Is early cord clamping, delayed cord clamping or cord milking best?

    Science.gov (United States)

    Vatansever, Binay; Demirel, Gamze; Ciler Eren, Elif; Erel, Ozcan; Neselioglu, Salim; Karavar, Hande Nur; Gundogdu, Semra; Ulfer, Gozde; Bahadir, Selcen; Tastekin, Ayhan

    2018-04-01

    To compare the antioxidant status of three cord clamping procedures (early clamping, delayed clamping and milking) by analyzing the thiol-disulfide balance. This randomized controlled study enrolled 189 term infants who were divided into three groups according to the cord clamping procedure: early clamping, delayed clamping and milking. Blood samples were collected from the umbilical arteries immediately after clamping, and the thiol/disulfide homeostasis was analyzed. The native and total thiol levels were significantly (p cord clamping group compared with the other two groups. The disulfide/total thiol ratio was significantly (p = .026) lower in the delayed cord clamping and milking groups compared with the early clamping groups. Early cord clamping causes the production of more disulfide bonds and lower thiol levels, indicating that oxidation reactions are increased in the early cord clamping procedure compared with the delayed cord clamping and milking procedures. The oxidant capacity is greater with early cord clamping than with delayed clamping or cord milking. Delayed cord clamping or milking are beneficial in neonatal care, and we suggest that they be performed routinely in all deliveries.

  5. Solution structure of an "open" E. coli Pol III clamp loader sliding clamp complex.

    Science.gov (United States)

    Tondnevis, Farzaneh; Weiss, Thomas M; Matsui, Tsutomu; Bloom, Linda B; McKenna, Robert

    2016-06-01

    Sliding clamps are opened and loaded onto primer template junctions by clamp loaders, and once loaded on DNA, confer processivity to replicative polymerases. Previously determined crystal structures of eukaryotic and T4 clamp loader-clamp complexes have captured the sliding clamps in either closed or only partially open interface conformations. In these solution structure studies, we have captured for the first time the clamp loader-sliding clamp complex from Escherichia coli using size exclusion chromatography coupled to small angle X-ray scattering (SEC-SAXS). The data suggests the sliding clamp is in an open conformation which is wide enough to permit duplex DNA binding. The data also provides information about spatial arrangement of the sliding clamp with respect to the clamp loader subunits and is compared to complex crystal structures determined from other organisms. Published by Elsevier Inc.

  6. Loading dynamics of a sliding DNA clamp.

    KAUST Repository

    Cho, Won-Ki

    2014-05-22

    Sliding DNA clamps are loaded at a ss/dsDNA junction by a clamp loader that depends on ATP binding for clamp opening. Sequential ATP hydrolysis results in closure of the clamp so that it completely encircles and diffuses on dsDNA. We followed events during loading of an E. coli β clamp in real time by using single-molecule FRET (smFRET). Three successive FRET states were retained for 0.3 s, 0.7 s, and 9 min: Hydrolysis of the first ATP molecule by the γ clamp loader resulted in closure of the clamp in 0.3 s, and after 0.7 s in the closed conformation, the clamp was released to diffuse on the dsDNA for at least 9 min. An additional single-molecule polarization study revealed that the interfacial domain of the clamp rotated in plane by approximately 8° during clamp closure. The single-molecule polarization and FRET studies thus revealed the real-time dynamics of the ATP-hydrolysis-dependent 3D conformational change of the β clamp during loading at a ss/dsDNA junction.

  7. Dynamics of Open DNA Sliding Clamps.

    Directory of Open Access Journals (Sweden)

    Aaron J Oakley

    Full Text Available A range of enzymes in DNA replication and repair bind to DNA-clamps: torus-shaped proteins that encircle double-stranded DNA and act as mobile tethers. Clamps from viruses (such as gp45 from the T4 bacteriophage and eukaryotes (PCNAs are homotrimers, each protomer containing two repeats of the DNA-clamp motif, while bacterial clamps (pol III β are homodimers, each protomer containing three DNA-clamp motifs. Clamps need to be flexible enough to allow opening and loading onto primed DNA by clamp loader complexes. Equilibrium and steered molecular dynamics simulations have been used to study DNA-clamp conformation in open and closed forms. The E. coli and PCNA clamps appear to prefer closed, planar conformations. Remarkably, gp45 appears to prefer an open right-handed spiral conformation in solution, in agreement with previously reported biophysical data. The structural preferences of DNA clamps in solution have implications for understanding the duty cycle of clamp-loaders.

  8. The RFC Clamp Loader: Structure and Function

    Science.gov (United States)

    O'Donnell, Mike

    2013-01-01

    The eukaryotic RFC clamp loader couples the energy of ATP hydrolysis to open and close the circular PCNA sliding clamp onto primed sites for use by DNA polymerases and repair factors. Structural studies reveal clamp loaders to be heteropentamers. Each subunit contains a region of homology to AAA+ proteins that defines two domains. The AAA+ domains form a right-handed spiral upon binding ATP. This spiral arrangement generates a DNA binding site within the center of RFC. DNA enters the central chamber through a gap between the AAA+ domains of two subunits. Specificity for a primed template junction is achieved by a third domain that blocks DNA, forcing it to bend sharply. Thus only DNA with a flexible joint can bind the central chamber. DNA entry also requires a slot in the PCNA clamp, which is opened upon binding the AAA+ domains of the clamp loader. ATP hydrolysis enables clamp closing and ejection of RFC, completing the clamp loading reaction. PMID:22918590

  9. Effects of glucagon-like peptide 1 on counterregulatory hormone responses, cognitive functions, and insulin secretion during hyperinsulinemic, stepped hypoglycemic clamp experiments in healthy volunteers

    DEFF Research Database (Denmark)

    Nauck, Michael A; Heimesaat, Markus M; Behle, Kai

    2002-01-01

    Glucagon-like peptide 1 (GLP-1) and analogues are being evaluated as a new therapeutic principle for the treatment of type 2 diabetes. GLP-1 suppresses glucagon secretion, which could lead to disturbances of hypoglycemia counterregulation. This has, however, not been tested. Nine healthy voluntee......, GLP-1 (1.2 pmol x kg(-1) x min(-1)) was administered i.v. (steady-state concentration, approximately 125 pmol/liter); on the other occasion, NaCl was administered as placebo. Glucagon, cortisol, GH (immunoassays), and catecholamines (radioenzymatic assay) were determined, autonomous.......97). The other counterregulatory hormones and autonomic or neuroglucopenic symptom scores increased, and cognitive functions decreased with decreasing glucose concentrations, but there were no significant differences comparing experiments with GLP-1 or placebo, except for a significant reduction of GH responses...

  10. Dynamics of insulin signalling in liver during hyperinsulinemic euglycaemic clamp conditions in vivo and the effects of high-fat feeding in male mice

    NARCIS (Netherlands)

    Korsheninnikova, E.; Voshol, P.J.; Baan, B.; Zon, G.C.M. van der; Havekes, L.M.; Romijn, J.A.; Maassen, J.A.; Ouwens, D.M.

    2007-01-01

    Insulin is an important regulator of hepatic carbohydrate, lipid, and protein metabolism, and the regulation of these processes by insulin is disturbed under conditions of insulin resistance and type 2 diabetes. Despite these alterations, the impact of insulin resistance on insulin signalling in the

  11. Dynamics of insulin signalling in liver during hyperinsulinemic euglycaemic clamp conditions in vivo and the effects of high-fat feeding in male mice.

    Science.gov (United States)

    Korsheninnikova, E; Voshol, P J; Baan, B; van der Zon, G C M; Havekes, L M; Romijn, J A; Maassen, J A; Ouwens, D M

    2007-01-01

    Insulin is an important regulator of hepatic carbohydrate, lipid, and protein metabolism, and the regulation of these processes by insulin is disturbed under conditions of insulin resistance and type 2 diabetes. Despite these alterations, the impact of insulin resistance on insulin signalling in the liver is not well defined. Variations in time and dose of insulin stimulation as well as plasma glucose levels may underlie this. The present study aimed at determining the dynamics of activation of hepatic insulin signalling in vivo at insulin concentrations resembling those achieved after a meal, and addressing the effects of high-fat feeding. An unexpected finding of this study was the biphasic activation pattern of the IRS-PI3K-PKB/Akt pathway. Our findings indicate that the first burst of activation contributes to regulation of glucose metabolism. The physiological function of the second peak is still unknown, but may involve regulation of protein synthesis. Finally, high-fat feeding caused hepatic insulin resistance, as illustrated by a reduced suppression of hepatic glucose production. A sustained increased phosphorylation of the serine/threonine kinases p70S6kinase and Jun N-terminal kinase in the absence of insulin may underlie the abrogated phosphorylation of the IRS proteins and their downstream targets.

  12. Pancreatic arterial calcium stimulation in the diagnosis and localisation of persistent hyperinsulinemic hypoglycaemia of infancy

    Energy Technology Data Exchange (ETDEWEB)

    Chigot, V.; Brunelle, F. [Department of Radiology, Hopital des Enfants Malades, Paris (France); Lonlay, P. de; Nassogne, M.-C.; Delagne, V.; Saudubray, J.-M. [Dept. of Paediatrics, Hopital des Enfants Malades, Paris (France); Laborde, K. [Dept. of Biology, Hopital des Enfants Malades, Paris (France); Fournet, J.-C. [Dept. of Pathology, Hopital des Enfants Malades, Paris (France); Nihoul-Fekete, C. [Department of Surgery, Hopital des Enfants Malades, Paris (France)

    2001-09-01

    Persistent hyperinsulinemic hypoglycaemia of infancy (PHHI) is often resistant to medical therapy. Surgery is therefore necessary. It is due to focal adenomatous islet-cell hyperplasia treatable by partial pancreatectomy, or diffuse beta-cell hyperfunction, which requires near-total pancreatectomy. Pancreatic venous sampling (PVS) is the reference technique for the preoperative diagnosis and localization of focal forms of PHHI in the pancreas. However, hypoglycaemia is necessary to analyse the results and PVS is technically challenging. Pancreatic arterial calcium stimulation (PACS) is technically easier and does not require hypoglycaemia. To study the accuracy in the diagnosis and localization of PHHI. Materials and methods: PACS was performed in 12 patients and correlated with histology. The accuracy of PACS is poor in diffuse lesions since only two of six cases were correctly identified by this test. Five of six focal lesions were correctly recognized and located. PACS is less accurate than PVS in PHHI. Currently, it should be performed only when PVS fails. (orig.)

  13. Poor pregnancy outcome after octreotide treatment during pregnancy for familial hyperinsulinemic hypoglycemia

    DEFF Research Database (Denmark)

    Skajaa, Gitte O; Mathiesen, Elisabeth R; Iyore, Elisabeth

    2014-01-01

    growth. At birth the girl was small for gestational age. She was admitted to the neonatal special care unit due to low blood glucose and intravenous glucose and early feeding was initiated. One day old, her condition deteriorated with signs of an abdominal catastrophe indicating necrotizing enterocolitis...... successfully. One child carried the mutation and was small for gestational age at birth while the other child did not carry the mutation and had normal birth weight. CONCLUSION: In a woman with late familial hyperinsulinemic hypoglycemia octreotide was given during the first four pregnancies resulting in 2...... cases of early termination of pregnancy on parental request and 2 cases of inappropriate fetal growth and unviable outcome. The following two pregnancies treated with diet only had a successful outcome....

  14. A novel syndrome of autosomal-dominant hyperinsulinemic hypoglycemia linked to a mutation in the human insulin receptor gene

    DEFF Research Database (Denmark)

    Højlund, Kurt; Hansen, Torben; Lajer, Maria

    2004-01-01

    a missense mutation (Arg1174Gln) in the tyrosine kinase domain of the insulin receptor gene that cosegregated with the disease phenotype (logarithm of odds [LOD] score 3.21). In conclusion, we report a novel syndrome of autosomal-dominant hyperinsulinemic hypoglycemia. The findings demonstrate...

  15. Arterial clamping leads to stenosis at clamp sites after femoropopliteal bypass surgery.

    Science.gov (United States)

    Vriens, Bianca H R; Pol, Robert A; Hulsebos, Robin G; van Det, Rob J; van der Palen, Job; Zeebregts, Clark J; Geelkerken, Robert H

    2015-09-01

    To date, the incidence and clinical relevance of arterial stenosis at clamp sites after femoropopliteal bypass surgery is unknown. Ninety-four patients underwent a femoropopliteal bypass in which the arterial inflow and outflow clamp sites were controlled by the Fogarty-Soft-Inlay clamp and marked with an hemoclip. The number of pre-existing atherosclerotic segments, clamp force, and clamp time were recorded and the occurrence of a stenosis at the clamp site was determined. After a mean follow-up of 83 months, a significant stenosis was confirmed at 23 of the 178 clamp sites (12.9%; 95% confidence interval 8.4 to 18.8). The mean number of pre-existing atherosclerotic segments (P = .28) and the mean clamp force (P = .55) was similar between the groups with and without a stenosis. There was a significant difference regarding clamp time between the group with and without a stenosis (38 minutes and 26 minutes, P = .001). Arterial clamping, even with the Fogarty-Soft-Inlay clamp, can lead to clamp stenosis and seems to be related to the duration of clamping, but not to pre-existent atherosclerotic burden. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. To clamp or not to clamp? Long-term functional outcomes for elective off-clamp laparoscopic partial nephrectomy.

    Science.gov (United States)

    Shah, Paras H; George, Arvin K; Moreira, Daniel M; Alom, Manaf; Okhunov, Zhamshid; Salami, Simpa; Waingankar, Nikhil; Schwartz, Michael J; Vira, Manish A; Richstone, Lee; Kavoussi, Louis R

    2016-02-01

    To evaluate whether elective off-clamp laparoscopic partial nephrectomy (LPN) affords long-term renal functional benefit compared with the on-clamp approach. This is a retrospective review of patients who underwent elective LPN between 2006 and 2011. Patients were followed longitudinally for up to 5 years. In all, 315 patients with radiographic evidence of a solitary renal mass and normal-appearing contralateral kidney underwent elective LPN; 209 were performed on-clamp vs 106 off-clamp. One patient who required conversion from LPN to open PN was excluded from the study. Additionally, four patients in the on-clamp cohort who underwent subsequent radical nephrectomy for local-regional recurrence were excluded from longitudinal functional evaluation after their procedure. The primary objective was to evaluate differences in postoperative estimated glomerular filtration rate (eGFR) between hilar clamping groups. Subgroup analyses were performed for patients with clamp times >30 min and those with baseline renal insufficiency (eGFR clamp and off-clamp cohorts (80.7 vs 84.1 mL/min/1.73m(2) , P > 0.05). Univariable and multivariable analyses did not show significant differences in postoperative eGFR between both groups among all-comers, those with clamp times >30 min, and patients with baseline renal insufficiency. Risk of chronic kidney disease was not diminished by the off-clamp approach with up to 5 years of follow-up. Progressive recovery of renal function after hilar clamping in the elective setting eclipses short-term functional benefit achieved with off-clamp LPN by 6 months; there was no significant difference in eGFR or the percentage incidence of chronic kidney disease between the on-clamp and off-clamp cohorts with up to 5 years follow-up. As such, eliminating transient ischaemia during elective LPN does not confer clinical benefit. © 2015 The Authors BJU International © 2015 BJU International Published by John Wiley & Sons Ltd.

  17. Comparison of the physiological relevance of systemic vs. portal insulin delivery to evaluate whole body glucose flux during an insulin clamp.

    Science.gov (United States)

    Farmer, Tiffany D; Jenkins, Erin C; O'Brien, Tracy P; McCoy, Gregory A; Havlik, Allison E; Nass, Erik R; Nicholson, Wendell E; Printz, Richard L; Shiota, Masakazu

    2015-02-01

    To understand the underlying pathology of metabolic diseases, such as diabetes, an accurate determination of whole body glucose flux needs to be made by a method that maintains key physiological features. One such feature is a positive differential in insulin concentration between the portal venous and systemic arterial circulation (P/S-IG). P/S-IG during the determination of the relative contribution of liver and extra-liver tissues/organs to whole body glucose flux during an insulin clamp with either systemic (SID) or portal (PID) insulin delivery was examined with insulin infusion rates of 1, 2, and 5 mU·kg(-1)·min(-1) under either euglycemic or hyperglycemic conditions in 6-h-fasted conscious normal rats. A P/S-IG was initially determined with endogenous insulin secretion to exist with a value of 2.07. During an insulin clamp, while inhibiting endogenous insulin secretion by somatostatin, P/S-IG remained at 2.2 with PID, whereas, P/S-IG disappeared completely with SID, which exhibited higher arterial and lower portal insulin levels compared with PID. Consequently, glucose disappearance rates and muscle glycogen synthetic rates were higher, but suppression of endogenous glucose production and liver glycogen synthetic rates were lower with SID compared with PID. When the insulin clamp was performed with SID at 2 and 5 mU·kg(-1)·min(-1) without managing endogenous insulin secretion under euglycemic but not hyperglycemic conditions, endogenous insulin secretion was completely suppressed with SID, and the P/S-IG disappeared. Thus, compared with PID, an insulin clamp with SID underestimates the contribution of liver in response to insulin to whole body glucose flux. Copyright © 2015 the American Physiological Society.

  18. Loading dynamics of a sliding DNA clamp.

    Science.gov (United States)

    Cho, Won-Ki; Jergic, Slobodan; Kim, Daehyung; Dixon, Nicholas E; Lee, Jong-Bong

    2014-06-23

    Sliding DNA clamps are loaded at a ss/dsDNA junction by a clamp loader that depends on ATP binding for clamp opening. Sequential ATP hydrolysis results in closure of the clamp so that it completely encircles and diffuses on dsDNA. We followed events during loading of an E. coli β clamp in real time by using single-molecule FRET (smFRET). Three successive FRET states were retained for 0.3 s, 0.7 s, and 9 min: Hydrolysis of the first ATP molecule by the γ clamp loader resulted in closure of the clamp in 0.3 s, and after 0.7 s in the closed conformation, the clamp was released to diffuse on the dsDNA for at least 9 min. An additional single-molecule polarization study revealed that the interfacial domain of the clamp rotated in plane by approximately 8° during clamp closure. The single-molecule polarization and FRET studies thus revealed the real-time dynamics of the ATP-hydrolysis-dependent 3D conformational change of the β clamp during loading at a ss/dsDNA junction. © 2014 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

  19. Sub-nanometer stable precision MEMS clamping mechanism maintaining clamp force unpowered for TEM application

    NARCIS (Netherlands)

    Brouwer, Dannis Michel; de Jong, B.R.; Soemers, Herman; van Dijk, Johannes

    2006-01-01

    A design is presented for a relatively large force (0.5 mN) high-precision MEMS clamping mechanism. The clamp is a part of a MEMS transmission electron microscope (TEM) sample manipulator, which needs to be fixed unpowered once positioned. The elastic deformation of the clamp suspension has been

  20. Electrostatic clamp manufactured by novel method

    NARCIS (Netherlands)

    Sligte, E. te; Storm, A.; Koster, N.B.

    2011-01-01

    Electrostatic clamps (ESCs), used in reticle and wafer handling, are presently manufactured using glass bonding and polishing technologies. We present a patented alternative concept to this process, relying on coating and etching processes rather than bonding. We manufactured a first prototype clamp

  1. Clamp and grout systems for structural repair

    Energy Technology Data Exchange (ETDEWEB)

    Lalani, M.

    1984-10-01

    Steel offshore platforms in the North Sea and elsewhere have required repair or strengthening, opening up a new and important aspect of offshore engineering. A clamp and grout system for structured repair is described. Over 150 clamped and grouted repair systems are now in service in the North Sea and elsewhere.

  2. Dumping syndrome: an unusual cause of severe hyperinsulinemic hypoglycemia in neurologically impaired children with gastrostomy.

    Science.gov (United States)

    Bizzarri, C; Cervoni, M; Crea, F; Cutrera, R; Schiavino, A; Schiaffini, R; Cappa, M

    2011-02-01

    This paper describes severe hyperinsulinemic hypoglycemia during bolus enteral feeding in two neurologically impaired children. Both children were affected by dysphagia with swallowing difficulties; caloric intake was inadequate. For these reasons, percutaneous endoscopic gastrostomy had been positioned during the first months of life. In one patient due to persisting vomiting, after a few months, a gastrojejunal tube (PEG-J) was inserted. Hypoglycemia was revealed by routine blood tests, without evidence of specific symptoms. Continuous subcutaneous glucose monitoring showed wide glucose excursions, ranging from hypoglycemia to hyperglycemia. Extremely high levels of insulin were detected at the time of hypoglycemia. A diagnosis of dumping syndrome (DS) was suspected in both children. In the child with PEG, the tip of the gastrostomy catheter was found to be lying in the bulbus duodeni. Once this had been pulled back, hypoglycemic episodes disappeared. The child with PEG-J needed continuous enteral feeding to reach a normal glucose balance. DS is a relatively common complication in children with gastrostomy, but extremely irregular glucose levels, ranging from hypoglycemia to hyperglycemia, and increased insulin secretion had not been previously demonstrated. The incidence of DS is probably underestimated in children receiving enteral feeding for neurological impairment. In these patients intensive monitoring of blood glucose levels should be performed to calibrate meals. Repeated underestimated hypoglycemic episodes could worsen neurological damage and cause a deterioration in clinical conditions.

  3. Clamp force and alignment checking device

    Science.gov (United States)

    Spicer, John Patrick; Cai, Wayne W.; Chakraborty, Debejyo; Mink, Keith

    2017-04-11

    A check fixture measures a total clamp force applied by a welder device. The welder device includes a welding horn having a plurality of weld pads and welding anvil having a plurality of weld pads. The check fixture includes a base member operatively supporting a plurality of force sensors. The base member and the force sensors are received between the weld pads of the welding horn and the anvil pads of the welding anvil. Each force sensor is configured to measure an individual clamp force applied thereto by corresponding weld and anvil pads when the base member is received between the welding horn and the welding anvil and the welder device is in the clamped position. The individual clamp forces are used to determine whether the weld and/or anvil pads are worn or misaligned.

  4. Assessment of Nifedipine Therapy in Hyperinsulinemic Hypoglycemia due to Mutations in the ABCC8 Gene.

    Science.gov (United States)

    Güemes, Maria; Shah, Pratik; Silvera, Shavel; Morgan, Kate; Gilbert, Clare; Hinchey, Louise; Hussain, Khalid

    2017-03-01

    Previous case reports have documented the effectiveness of l-type calcium channel blockers (such as nifedipine and verapamil) for treating different forms of hyperinsulinemic hypoglycemia (HH). To systematically assess the glycemic response to nifedipine therapy in 11 patients with HH due to mutations in the ABCC8 gene. Dose escalation of nifedipine therapy. Eleven children who were inpatients at a tertiary hospital and had diazoxide unresponsive HH due to mutations in the ABCC8 gene. Nifedipine was administered orally at an escalating dose up to a maximum of 2.5 mg/kg/d. Improvement in glycemic control, avoidance of hypoglycemic episodes, reduction of intravenous glucose infusion, and reduction in the requirements of other medical therapies. The median age of the patients was 0.44 years (range 0.14 to 3.77). The ABCC8 gene mutations were homozygous in 3 cases, paternally inherited heterozygous in 4, and compound heterozygous in 4. 18F-DOPA PET/CT scan demonstrated a focal lesion in 2 cases and the rest were diffuse HH disease. One subject had nifedipine as monotherapy, whereas the rest had it in combination with octreotide/glucagon/diazoxide or cornstarch. After a median of 6.5 (3 to 23) days of maximal (2.5 mg/kg/d) dose of nifedipine therapy, none of the patients showed any improvement in glycemic control and patients continued to have hypoglycemic episodes. HH due to mutations in the ABCC8 gene does not respond to nifedipine therapy. Mutations in the KATP channel genes might render the l-type calcium channel ineffective to therapy with nifedipine.

  5. Use of the Satinsky clamp for hilar clamping during robotic partial nephrectomy: indications, technique, and multi-center outcomes.

    Science.gov (United States)

    Abdullah, Newaj; Rahbar, Haider; Barod, Ravi; Dalela, Deepansh; Larson, Jeff; Johnson, Michael; Mass, Alon; Zargar, Homayoun; Kaouk, Jihad; Allaf, Mohamad; Bhayani, Sam; Stifelman, Michael; Rogers, Craig

    2017-03-01

    A Satinsky clamp may be a backup option for hilar clamping during robotic partial nephrectomy (RPN) if there are challenges with application of bulldog clamps, but there are potential safety concerns. We evaluate outcomes of RPN using Satinsky vs. bulldog clamps, and provide tips for safe use of the Satinsky as a backup option. Using a multi-center database, we identified 1073 patients who underwent RPN between 2006 and 2013, and had information available about method of hilar clamping (bulldog clamp vs. Satinsky clamp). Patient baseline characteristics, tumor features, and perioperative outcomes were compared between the Satinsky and bulldog clamp groups. A Satinsky clamp was used for hilar clamping in 94 (8.8 %) RPN cases, and bulldog clamps were used in 979 (91.2 %) cases. The use of a Satinsky clamp was associated with greater operative time (198 vs. 175 min, p clamp group, but none were directly related to the Satinsky clamp. On multivariable analysis, the use of the Satinsky clamp was not associated with increase in intraoperative or Clavien ≥3 postoperative complications, positive surgical margin rate or percentage change in estimated glomerular filtration rate. A Satinsky clamp can be a backup option for hilar clamping during challenging RPN cases, but requires careful technique, and was rarely necessary.

  6. 21 CFR 882.4460 - Neurosurgical head holder (skull clamp).

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Neurosurgical head holder (skull clamp). 882.4460... holder (skull clamp). (a) Identification. A neurosurgical head holder (skull clamp) is a device used to clamp the patient's skull to hold head and neck in a particular position during surgical procedures. (b...

  7. Arterial clamping leads to stenosis at clamp sites after femoropopliteal bypass surgery

    NARCIS (Netherlands)

    Vriens, Bianca H. R.; Pol, Robert A.; Hulsebos, Robin G.; van Det, Rob J.; van der Palen, Job; Zeebregts, Clark J.; Geelkerken, Robert H.

    BACKGROUND: To date, the incidence and clinical relevance of arterial stenosis at clamp sites after femoropopliteal bypass surgery is unknown. METHODS: Ninety-four patients underwent a femoropopliteal bypass in which the arterial inflow and outflow clamp sites were controlled by the

  8. Arterial clamping leads to stenosis at clamp sites after femoropopliteal bypass surgery

    NARCIS (Netherlands)

    Vriens, Bianca H.R.; Pol, Robert A.; Hulsebos, Robin G.; van Det, Rob J.; van der Palen, Jacobus Adrianus Maria; Zeebregts, Clark J.; Geelkerken, Robert H.

    2015-01-01

    Background To date, the incidence and clinical relevance of arterial stenosis at clamp sites after femoropopliteal bypass surgery is unknown. Methods Ninety-four patients underwent a femoropopliteal bypass in which the arterial inflow and outflow clamp sites were controlled by the Fogarty-Soft-Inlay

  9. Insulin Plays a Permissive Role for the Vasoactive Effect of GIP Regulating Adipose Tissue Metabolism in Humans

    DEFF Research Database (Denmark)

    Asmar, Meena; Simonsen, Lene; Asmar, Ali

    2016-01-01

    of insulin for the vasoactive effect of GIP in adipose tissue metabolism and whether the vasodilatory effect of GIP is dependent on C-peptide. METHODS: Six lean healthy subjects were studied. The sc abdominal adipose tissue metabolism was assessed by Fick's principle during GIP infusion (1.5 pmol....../kg/min) in combination with 1) euglycemic-high insulinemic clamp (Eugluc-Hiinsu), raising plasma insulin concentrations to postprandial levels, 2) hyperglycemic-euinsulinemic clamp (Hygluc-Euinsu), and 3) hyperglycemic-hyperinsulinemic clamp, raising plasma insulin concentrations to supraphysiological levels. During...... the hyperglycemic clamps, endogenous insulin and C-peptide secretion were inhibited by infusion of the somatostatin analogue octreotide. RESULTS: During GIP infusion, Eugluc-Hiinsu, and hyperglycemic-hyperinsulinemic clamps, sc abdominal adipose tissue blood flow (ATBF) was similar and increased from 2.1 ± 0...

  10. Piezoresistive cantilever force-clamp system

    Energy Technology Data Exchange (ETDEWEB)

    Park, Sung-Jin; Petzold, Bryan C.; Pruitt, Beth L. [Department of Mechanical Engineering, Stanford University, Stanford, California 94305 (United States); Goodman, Miriam B. [Department of Molecular and Cellular Physiology, Stanford University, Stanford, California 94305 (United States)

    2011-04-15

    We present a microelectromechanical device-based tool, namely, a force-clamp system that sets or ''clamps'' the scaled force and can apply designed loading profiles (e.g., constant, sinusoidal) of a desired magnitude. The system implements a piezoresistive cantilever as a force sensor and the built-in capacitive sensor of a piezoelectric actuator as a displacement sensor, such that sample indentation depth can be directly calculated from the force and displacement signals. A programmable real-time controller operating at 100 kHz feedback calculates the driving voltage of the actuator. The system has two distinct modes: a force-clamp mode that controls the force applied to a sample and a displacement-clamp mode that controls the moving distance of the actuator. We demonstrate that the system has a large dynamic range (sub-nN up to tens of {mu}N force and nm up to tens of {mu}m displacement) in both air and water, and excellent dynamic response (fast response time, <2 ms and large bandwidth, 1 Hz up to 1 kHz). In addition, the system has been specifically designed to be integrated with other instruments such as a microscope with patch-clamp electronics. We demonstrate the capabilities of the system by using it to calibrate the stiffness and sensitivity of an electrostatic actuator and to measure the mechanics of a living, freely moving Caenorhabditis elegans nematode.

  11. Whole-cell voltage clamp on skeletal muscle fibers with the silicone-clamp technique.

    Science.gov (United States)

    Lefebvre, Romain; Pouvreau, Sandrine; Collet, Claude; Allard, Bruno; Jacquemond, Vincent

    2014-01-01

    Control of membrane voltage and membrane current measurements are of critical importance for the study of numerous aspects of skeletal muscle physiology and pathophysiology. The silicone-clamp technique makes use of a conventional patch-clamp apparatus to achieve whole-cell voltage clamp of a restricted portion of a fully differentiated adult skeletal muscle fiber. The major part of an isolated muscle fiber is insulated from the extracellular medium with silicone grease and the tip of a single microelectrode connected to the amplifier is then inserted within the fiber through the silicone layer. The method is extremely easy to implement. It represents an alternative to the traditional vaseline-gap isolation and two or three microelectrodes voltage-clamp techniques. The present chapter reviews the benefits of the silicone-clamp technique and provides updated detailed insights into its practical implementation.

  12. IMPORTANT: Fluke is recalling Digital Clamp Meters

    CERN Multimedia

    2013-01-01

    Fluke is voluntarily recalling four models of Digital Clamp Meters: Fluke 373, 374, 375 and 376. If you own one of these clamp meters, please stop using it and send it back to Fluke for repair even if you have not experienced problems.   Description of the problem: "The printed circuit assembly may not be properly fastened to the test lead input jack. This may result in inaccurate voltage readings, including a low or no-voltage reading on a circuit energised with a hazardous voltage, presenting a shock, electrocution or thermal burn hazard." To determine if your clamp meter is affected by this recall notice, and for more information, click here.

  13. Kinetic analysis of PCNA clamp binding and release in the clamp loading reaction catalyzed by Saccharomyces cerevisiae replication factor C.

    Science.gov (United States)

    Marzahn, Melissa R; Hayner, Jaclyn N; Meyer, Jennifer A; Bloom, Linda B

    2015-01-01

    DNA polymerases require a sliding clamp to achieve processive DNA synthesis. The toroidal clamps are loaded onto DNA by clamp loaders, members of the AAA+family of ATPases. These enzymes utilize the energy of ATP binding and hydrolysis to perform a variety of cellular functions. In this study, a clamp loader-clamp binding assay was developed to measure the rates of ATP-dependent clamp binding and ATP-hydrolysis-dependent clamp release for the Saccharomyces cerevisiae clamp loader (RFC) and clamp (PCNA). Pre-steady-state kinetics of PCNA binding showed that although ATP binding to RFC increases affinity for PCNA, ATP binding rates and ATP-dependent conformational changes in RFC are fast relative to PCNA binding rates. Interestingly, RFC binds PCNA faster than the Escherichia coli γ complex clamp loader binds the β-clamp. In the process of loading clamps on DNA, RFC maintains contact with PCNA while PCNA closes, as the observed rate of PCNA closing is faster than the rate of PCNA release, precluding the possibility of an open clamp dissociating from DNA. Rates of clamp closing and release are not dependent on the rate of the DNA binding step and are also slower than reported rates of ATP hydrolysis, showing that these rates reflect unique intramolecular reaction steps in the clamp loading pathway. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Fibroblast growth factor-21 is induced in human skeletal muscles by hyperinsulinemia

    DEFF Research Database (Denmark)

    Hojman, Pernille; Pedersen, Maria; Nielsen, Anders Rinnov

    2009-01-01

    DESIGN AND METHODS: We studied muscular FGF-21 expression and plasma FGF-21 after acute insulin stimulation in young healthy men during a hyperinsulinemic-euglycemic clamp. Furthermore, we investigated systemic levels and muscle FGF-21 expression in humans with or without insulin resistance and chronic...... elevated insulin. RESULTS: FGF-21 was barely detectable in young healthy men before insulin infusion. After 3 or 4 h of insulin infusion during a hyperinsulinemic-euglycemic clamp, muscular FGF-21 expression increased significantly. Plasma FGF-21 followed the same pattern. In individuals with chronic...... elevated insulin, muscular FGF-21 expression was associated with hyperinsulinemia in men but not in women. In plasma, hyperinsulinemia and fasting glucose were positively associated with plasma FGF-21 while plasma FGF-21 correlated negatively with HDL cholesterol. No associations between muscle and plasma...

  15. Clamp loader ATPases and the evolution of DNA replication machinery

    Science.gov (United States)

    2012-01-01

    Clamp loaders are pentameric ATPases of the AAA+ family that operate to ensure processive DNA replication. They do so by loading onto DNA the ring-shaped sliding clamps that tether the polymerase to the DNA. Structural and biochemical analysis of clamp loaders has shown how, despite differences in composition across different branches of life, all clamp loaders undergo the same concerted conformational transformations, which generate a binding surface for the open clamp and an internal spiral chamber into which the DNA at the replication fork can slide, triggering ATP hydrolysis, release of the clamp loader, and closure of the clamp round the DNA. We review here the current understanding of the clamp loader mechanism and discuss the implications of the differences between clamp loaders from the different branches of life. PMID:22520345

  16. Clamp loader ATPases and the evolution of DNA replication machinery

    Directory of Open Access Journals (Sweden)

    Kelch Brian A

    2012-04-01

    Full Text Available Abstract Clamp loaders are pentameric ATPases of the AAA+ family that operate to ensure processive DNA replication. They do so by loading onto DNA the ring-shaped sliding clamps that tether the polymerase to the DNA. Structural and biochemical analysis of clamp loaders has shown how, despite differences in composition across different branches of life, all clamp loaders undergo the same concerted conformational transformations, which generate a binding surface for the open clamp and an internal spiral chamber into which the DNA at the replication fork can slide, triggering ATP hydrolysis, release of the clamp loader, and closure of the clamp round the DNA. We review here the current understanding of the clamp loader mechanism and discuss the implications of the differences between clamp loaders from the different branches of life.

  17. The relationship of hyperinsulinemic state to left ventricular hypertrophy, microalbuminuria, and physical fitness in borderline and mild hypertension.

    Science.gov (United States)

    Tomiyama, H; Doba, N; Kushiro, T; Yamashita, M; Kanmatsuse, K; Kajiwara, N; Yoshida, H; Hinohara, S

    1997-06-01

    The relationship of the hyperinsulinemic state to left ventricular hypertrophy, left ventricular geometric patterns, microalbuminuria, and physical fitness were studied in 52 middle-aged unmedicated men with borderline and mild hypertension. Left ventricular mass index and relative wall thickness were assessed by echocardiography. Physical fitness was determined by symptom-limited maximal treadmill stress testings. The urinary concentration of microalbumin and C-peptide was measured in 24-h urine samples by radioimmunoassey. The 24-h urinary C-peptide excretion rate was correlated with left ventricular mass index (r = 0.46), relative wall thickness (r = 0.41), treadmill time (r = -0.35), normalized treadmill time (r = -0.52), systolic blood pressure at peak exercise (r = 0.29), and 24-h urinary microalbumin excretion (r = 0.48). Stepwise multiple regression analysis identified the left ventricular mass index, the 24-h urinary albumin excretion, and the normalized treadmill time as variables in the equation for the 24-h urinary C-peptide excretion. Thus, the hyperinsulinemic state is related to left ventricular hypertrophy, microalbuminuria, and impaired physical fitness in patients with borderline and mild hypertension.

  18. Inactivation of the first nucleotide-binding fold of the sulfonylurea receptor, and familial persistent hyperinsulinemic hypoglycemia of infancy

    Energy Technology Data Exchange (ETDEWEB)

    Thomas, P.M.; Wohllk, N.; Huang, E. [Univ. of Texas, Houston, TX (United States)] [and others

    1996-09-01

    Familial persistent hyperinsulinemic hypoglycemia of infancy is a disorder of glucose homeostasis and is characterized by unregulated insulin secretion and profound hypoglycemia. Loss-of-function mutations in the second nucleotide-binding fold of the sulfonylurea receptor, a subunit of the pancreatic-islet {beta}-cell ATP-dependent potassium channel, has been demonstrated to be causative for persistent hyperinsulinemic hypoglycemia of infancy. We now describe three additional mutations in the first nucleotide-binding fold of the sulfonylurea-receptor gene. One point mutation disrupts the highly conserved Walker A motif of the first nucleotide-binding-fold region. The other two mutations occur in noncoding sequences required for RNA processing and are predicted to disrupt the normal splicing pathway of the sulfonylurea-receptor mRNA precursor. These data suggest that both nucleotide-binding-fold regions of the sulfortylurea receptor are required for normal regulation of {beta}-cell ATP-dependent potassium channel activity and insulin secretion. 32 refs., 4 figs., 1 tab.

  19. The Effect of Near Total Pancreatectomy in Treatment of Hyperinsulinemic Hyperglycemic Infants Children’s Medical Center 2000

    Directory of Open Access Journals (Sweden)

    M Klantary

    2002-09-01

    Full Text Available Background: There are many surgical approaches for hyperinsulinemic hyperglycemic treatment. Subtotal pancreatectomy which has been used for many years, is abounded know due to high recurrence rate, and has been replaced by near total resection. This study focuses on “near total resection”. Materials and Mehods: In this study 15 cases of hyper insulinemic hyperglycemia patients which has been operated since year 1985 till 2000 in the Childrens Medical Center, has brought into consideration and described as a case series study. Results: From the patients, three cases were operated by subtotal pancreatomy and all of them have had recurrence. In the remaining twelve cases, the method used was different and “near subtotal pancreatomy” were used. The recurrence rate in this group was zero and there were not also any case of sepsis. Postoperative complications were rather low and the most common postoperative finding in these patients was transient hypoglycemia. Conclusion: Near total pancreatomy for hyperinsulinemic hypoglycemia which is implemented by an expert surgeon with low complication and high cure rate, can be considered as a safe technique with high cure rate and satisfactory results.

  20. Γ-source Neutral Point Clamped Inverter

    DEFF Research Database (Denmark)

    Mo, Wei; Loh, Poh Chiang; Blaabjerg, Frede

    Transformer based Z-source inverters are recently proposed to achieve promising buck-boost capability. They have improved higher buck-boost capability, smaller size and less components count over Z-source inverters. On the other hand, neutral point clamped inverters have less switching stress...

  1. Clamp and Gas Nozzle for TIG Welding

    Science.gov (United States)

    Gue, G. B.; Goller, H. L.

    1982-01-01

    Tool that combines clamp with gas nozzle is aid to tungsten/inert-gas (TIG) welding in hard-to-reach spots. Tool holds work to be welded while directing a stream of argon gas at weld joint, providing an oxygen-free environment for tungsten-arc welding.

  2. AN ACTIVE VALVE WITH A CLAMPED MEMBRANE

    DEFF Research Database (Denmark)

    2009-01-01

    An active valve for use e.g. in fluidic microsystems is provided, wherein the active valve comprises a membrane having at least one flow gate, arranged between a first and a second substantially rigid element. Adjusting means provides an adjustment of the clamping force on membrane arranged between...

  3. ANALYTICAL BENDING SOLUTION OF ALL CLAMPED ISOTROPIC ...

    African Journals Online (AJOL)

    ANALYTICAL BENDING SOLUTION OF ALL CLAMPED ISOTROPIC RECTANGULAR PLATE ON WINKLER’S FOUNDATION USING CHARACTERISTIC ... The PDF file you selected should load here if your Web browser has a PDF reader plug-in installed (for example, a recent version of Adobe Acrobat Reader).

  4. Comparison of the effects of barnidipine+losartan compared with telmisartan+hydrochlorothiazide on several parameters of insulin sensitivity in patients with hypertension and type 2 diabetes mellitus.

    Science.gov (United States)

    Derosa, Giuseppe; Querci, Fabrizio; Franzetti, Ivano; Dario Ragonesi, Pietro; D'Angelo, Angela; Maffioli, Pamela

    2015-10-01

    The aim of this study was to evaluate the effects of barnidipine+losartan compared with telmisartan+hydrochlorothiazide on several parameters of insulin sensitivity in patients with hypertension and type 2 diabetes mellitus. We enrolled 148 normocholesterolemic patients with mild-to-moderate hypertension and type 2 diabetes mellitus. Patients were treated with barnidipine, 20 mg day(-1), in combination with losartan, 100 mg day(-1), or with telmisartan+hydrochlorothiazide, 80/12.5 mg day(-1), for 6 months. We assessed blood pressure (BP) on a monthly basis; additionally, blood samples were collected to assess, at baseline and after 6 months, the following parameters: fasting plasma glucose; glycated hemoglobin; fasting plasma insulin; HOMA index; and some adipocytokines, such as adiponectin (ADN), resistin, leptin, visfatin and vaspin. Patients were also subjected to an euglycemic hyperinsulinemic clamp to assess the M value and glucose infusion rate to ascertain their insulin sensitivity. One hundred and forty-one patients completed the study. The BP was reduced in both groups, although the reduction was greater with barnidipine+losartan (Plosartan increased the M value and glucose infusion rate during the euglycemic hyperinsulinemic clamp (Plosartan (Plosartan compared with baseline (Plosartan were significantly better than those obtained with telmisartan+hydrochlorothiazide (Plosartan improved the insulin sensitivity, as assessed by an euglycemic hyperinsulinemic clamp, and improved some of the adipocytokines related to insulin resistance.

  5. Single molecule study of a processivity clamp sliding on DNA

    Energy Technology Data Exchange (ETDEWEB)

    Laurence, T A; Kwon, Y; Johnson, A; Hollars, C; O?Donnell, M; Camarero, J A; Barsky, D

    2007-07-05

    Using solution based single molecule spectroscopy, we study the motion of the polIII {beta}-subunit DNA sliding clamp ('{beta}-clamp') on DNA. Present in all cellular (and some viral) forms of life, DNA sliding clamps attach to polymerases and allow rapid, processive replication of DNA. In the absence of other proteins, the DNA sliding clamps are thought to 'freely slide' along the DNA; however, the abundance of positively charged residues along the inner surface may create favorable electrostatic contact with the highly negatively charged DNA. We have performed single-molecule measurements on a fluorescently labeled {beta}-clamp loaded onto freely diffusing plasmids annealed with fluorescently labeled primers of up to 90 bases. We find that the diffusion constant for 1D diffusion of the {beta}-clamp on DNA satisfies D {le} 10{sup -14} cm{sup 2}/s, much slower than the frictionless limit of D = 10{sup -10} cm{sup 2}/s. We find that the {beta} clamp remains at the 3-foot end in the presence of E. coli single-stranded binding protein (SSB), which would allow for a sliding clamp to wait for binding of the DNA polymerase. Replacement of SSB with Human RP-A eliminates this interaction; free movement of sliding clamp and poor binding of clamp loader to the junction allows sliding clamp to accumulate on DNA. This result implies that the clamp not only acts as a tether, but also a placeholder.

  6. Analysis of the Pallet Clamping Conditions in Work Positioning

    Directory of Open Access Journals (Sweden)

    L. A. Kolesnikov

    2016-01-01

    Full Text Available Pallets of automatic transfer lines serve as the technological and work-transfer equipment. However, their design is based on the general principles of designing machine accessories. Calculation of the basic design parameters of work based on their functioning features is fairly relevant.In pallet transfer movement over work positioning there is a mismatch between axes of the basic bushing of a pallet and the clamping locks of positioning. The paper identifies the factors influencing it, and defines the conditions to ensure the trouble-free clamping lock. The circular and rhombic clamping of pallet is simultaneous. Further, a clamping load is calculated from the pallet mass (together with the work-piece and the geometric parameters of the pallet clamping.The paper reveals three movement stages of the clamping lock in a vertical plane when setting the pallet in work positioning:- a clamping lock free play till it encounters with the location bushing;- a clamping lock play length when the lead-in chamfer contacts the bushing chamfer;- a clamping lock play length over the circular surface of bushing.The study of this process has allowed us to calculate the time of pallet clamping and dynamic loads acting on the clamping lock. These calculations made it possible to calculate the required diameter of the clamping lock. The presented calculations allow a reasonable assigning the main design parameters of work positioning for the pallet automatic lines.

  7. 21 CFR 882.5175 - Carotid artery clamp.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Carotid artery clamp. 882.5175 Section 882.5175...) MEDICAL DEVICES NEUROLOGICAL DEVICES Neurological Therapeutic Devices § 882.5175 Carotid artery clamp. (a) Identification. A carotid artery clamp is a device that is surgically placed around a patient's carotid artery...

  8. 21 CFR 876.5160 - Urological clamp for males.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Urological clamp for males. 876.5160 Section 876...) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5160 Urological clamp for males. (a) Identification. A urological clamp for males is a device used to close the urethra of a male to...

  9. Multilevel-clamped multilevel converters (MLC2)

    DEFF Research Database (Denmark)

    Rodriguez, Pedro; Bellar, Maria Dias; Muñoz-Aguliar, Raúl Santiago

    2012-01-01

    This letter introduces a new series of multilevel (ML) converters based on the ML clamping concept. By applying this technique, a ML clamping unit (MCU) conveys additional levels for synthesizing the output waveforms of a diode-clampedML dc– ac power converter. The basic building block of the ML ...

  10. High-speed dynamic-clamp interface

    Science.gov (United States)

    Yang, Yang; Adowski, Timothy; Ramamurthy, Bina; Neef, Andreas

    2015-01-01

    The dynamic-clamp technique is highly useful for mimicking synaptic or voltage-gated conductances. However, its use remains rare in part because there are few systems, and they can be expensive and difficult for less-experienced programmers to implement. Furthermore, some conductances (such as sodium channels) can be quite rapid or may have complex voltage sensitivity, so high speeds are necessary. To address these issues, we have developed a new interface that uses a common personal computer platform with National Instruments data acquisition and WaveMetrics IGOR to provide a simple user interface. This dynamic clamp implements leak and linear synaptic conductances as well as a voltage-dependent synaptic conductance and kinetic channel conductances based on Hodgkin-Huxley or Markov models. The speed of the system can be assayed using a testing mode, and currently speeds of >100 kHz (10 μs per cycle) are achievable with short latency and little jitter. PMID:25632075

  11. Towards a Dynamic Clamp for Neurochemical Modalities

    Directory of Open Access Journals (Sweden)

    Catalina Maria Rivera

    2015-05-01

    Full Text Available The classic dynamic clamp technique uses a real-time electrical interface between living cells and neural simulations in order to investigate hypotheses about neural function and structure. One of the acknowledged drawbacks of that technique is the limited control of the cells’ chemical microenvironment. In this manuscript, we use a novel combination of nanosensor and microfluidic technology and microfluidic and neural simulations to add sensing and control of chemical concentrations to the dynamic clamp technique. Specifically, we use a microfluidic lab-on-a-chip to generate distinct chemical concentration gradients (ions or neuromodulators, to register the concentrations with embedded nanosensors and use the processed signals as an input to simulations of a neural cell. The ultimate goal of this project is to close the loop and provide sensor signals to the microfluidic lab-on-a-chip to mimic the interaction of the simulated cell with other cells in its chemical environment.

  12. A Dynamic Clamp on Every Rig.

    Science.gov (United States)

    Desai, Niraj S; Gray, Richard; Johnston, Daniel

    2017-01-01

    The dynamic clamp should be a standard part of every cellular electrophysiologist's toolbox. That it is not, even 25 years after its introduction, comes down to three issues: money, the disruption that adding dynamic clamp to an existing electrophysiology rig entails, and the technical prowess required of experimenters. These have been valid and limiting issues in the past, but no longer. Technological advances associated with the so-called maker movement render them moot. We demonstrate this by implementing a fast (∼100 kHz) dynamic clamp system using an inexpensive microcontroller (Teensy 3.6). The overall cost of the system is less than USD$100, and assembling it requires no prior electronics experience. Modifying it-for example, to add Hodgkin-Huxley-style conductances-requires no prior programming experience. The system works together with existing electrophysiology data acquisition systems (for Macintosh, Windows, and Linux); it does not attempt to supplant them. Moreover, the process of assembling, modifying, and using the system constitutes a useful pedagogical exercise for students and researchers with no background but an interest in electronics and programming. We demonstrate the system's utility by implementing conductances as fast as a transient sodium conductance and as complex as the Ornstein-Uhlenbeck conductances of the "point conductance" model of synaptic background activity.

  13. Hyperinsulinemic hypoglycemia in Beckwith-Wiedemann syndrome due to defects in the function of pancreatic beta-cell adenosine triphosphate-sensitive potassium channels

    DEFF Research Database (Denmark)

    Hussain, K; Cosgrove, K E; Shepherd, R M

    2005-01-01

    Beckwith-Wiedemann syndrome (BWS) is a congenital overgrowth syndrome that is clinically and genetically heterogeneous. Hyperinsulinemic hypoglycemia occurs in about 50% of children with BWS and, in the majority of infants, it resolves spontaneously. However, in a small group of patients the hypo...

  14. Dental dam clamp adaptation method on carved gypsum cast.

    Science.gov (United States)

    Cazacu, N C E

    2014-01-01

    Dental Dam is the safest and most efficient isolation technique in endodontics and restorative dentistry, but it also used in esthetics, orthodontics, prosthetics, pedodontics and periodontology (for teeth immobilization). While in most cases the standard clamps are efficient, in some clinical situations clamp adaptation is mandatory in order to assure a tight contact on the tooth. The purpose of this study is to list the elements of the clamp, which should be modified in order to assure a secure constriction of the clamp on the anchor tooth, by using the carved gypsum cast method. 100 patients were examined, diagnosed and treated for various diagnoses like simple decay, gangrene, chronic apical periodontitis, and endodontic retreatments. The clamps used in this study were produced by Hu-Friedy, Hygenic, KKD, SDI, Hager & Werken. In 10 cases, the anchor tooth did not provide enough stability to the standard clamp--as provided by the producer. Therefore, we have done some adjustments to some of the elements of the clamp: the arch, the wings, the plateau, the active area, and the contact points. In 6 cases, major clamp adaptations on carved gypsum cast were imperative. The classic clamps cannot provide a grip to be enough in all the clinical cases due to the huge variety and position and implantation of the anchor teeth. Therefore, in such situations, the clamps should be adapted in order to provide stability and assure the safe isolation during the treatment. The modified clamps will be useful in similar cases, so they must be kept.

  15. Unequal pressure distribution along the jaws of currently available vascular clamps: do we need a new aortic clamp?

    Science.gov (United States)

    Rylski, Bartosz; Schmid, Claudius; Beyersdorf, Friedhelm; Kari, Fabian Alexander; Kondov, Stoyan; Lutz, Lisa; Werner, Martin; Czerny, Martin; Siepe, Matthias

    2016-06-01

    The pressure along vascular clamp jaws may be unequally distributed, with greater pressure near the clamp hinge than at its top. Such unequal pressure distribution may cause aortic injury, especially in large aortas. We evaluated pressure distribution along different currently availably clamp jaws. Seven descending thoracic aortas from pigs (diameter 2.0-3.0 cm) were plainly dissected and all side arteries closed. Aortas were filled up with water and cross-clamped. The pressure inside the aorta was raised to 100 mmHg and the aorta was clamped so tightly that no water exited from the distal aortic end. Each aorta was clamped seven times at different sites with the following clamps: DeBakey, Satinsky, femoral, iliac, Chitwood, angled handle Fogarty and straight handle Fogarty. The pressure along the clamp jaws was measured with a pressure-detecting film placed between the clamp jaws and aorta. The collagen-fibre disorganization was examined in haemotoxylin-eosin- and Elastica van Gieson-stained tissue samples. The DeBakey clamp revealed the lowest maximum pressure along the clamp jaws after complete aortic occlusion (1.43 ± 0.49 MPa), whereas the Chitwood clamp's pressure was the highest (3.26 ± 1.93 MPa, P clamp displayed the lowest difference between maximum pressures across the jaws (33%), with the greatest difference measured in the iliac (72%) and Chitwood (66%) clamps. The highest collagen-fibre disorganization score was observed in the proximal-to-the-clamp-hinge quartile after clamping with the angled handle Fogarty (2.8 ± 0.4), straight handle Fogarty (2.3 ± 0.8) and Chitwood (2.3 ± 0.5) clamps. The pressure along clamp jaws is unequally distributed in all the currently available vascular clamps. The Chitwood clamp is associated with the highest maximum pressure during complete aortic occlusion and with the most unequal pressure distribution along the jaws. © The Author 2015. Published by Oxford University Press on behalf of the European Association

  16. Whole-GUV patch-clamping.

    Science.gov (United States)

    Garten, Matthias; Mosgaard, Lars D; Bornschlögl, Thomas; Dieudonné, Stéphane; Bassereau, Patricia; Toombes, Gilman E S

    2017-01-10

    Studying how the membrane modulates ion channel and transporter activity is challenging because cells actively regulate membrane properties, whereas existing in vitro systems have limitations, such as residual solvent and unphysiologically high membrane tension. Cell-sized giant unilamellar vesicles (GUVs) would be ideal for in vitro electrophysiology, but efforts to measure the membrane current of intact GUVs have been unsuccessful. In this work, two challenges for obtaining the "whole-GUV" patch-clamp configuration were identified and resolved. First, unless the patch pipette and GUV pressures are precisely matched in the GUV-attached configuration, breaking the patch membrane also ruptures the GUV. Second, GUVs shrink irreversibly because the membrane/glass adhesion creating the high-resistance seal (>1 GΩ) continuously pulls membrane into the pipette. In contrast, for cell-derived giant plasma membrane vesicles (GPMVs), breaking the patch membrane allows the GPMV contents to passivate the pipette surface, thereby dynamically blocking membrane spreading in the whole-GMPV mode. To mimic this dynamic passivation mechanism, beta-casein was encapsulated into GUVs, yielding a stable, high-resistance, whole-GUV configuration for a range of membrane compositions. Specific membrane capacitance measurements confirmed that the membranes were truly solvent-free and that membrane tension could be controlled over a physiological range. Finally, the potential for ion transport studies was tested using the model ion channel, gramicidin, and voltage-clamp fluorometry measurements were performed with a voltage-dependent fluorophore/quencher pair. Whole-GUV patch-clamping allows ion transport and other voltage-dependent processes to be studied while controlling membrane composition, tension, and shape.

  17. The β Sliding Clamp Closes around DNA prior to Release by the Escherichia coli Clamp Loader γ Complex*

    Science.gov (United States)

    Hayner, Jaclyn N.; Bloom, Linda B.

    2013-01-01

    Escherichia coli γ complex clamp loader functions to load the β sliding clamp onto sites of DNA replication and repair. The clamp loader uses the energy of ATP binding and hydrolysis to drive conformational changes allowing for β binding and opening, DNA binding, and then release of the β·DNA complex. Although much work has been done studying the sliding clamp and clamp loader mechanism, kinetic analysis of the events following β·γ complex·DNA formation is not complete. Using fluorescent clamp closing and release assays, we show that β closing is faster than β release, indicating that γ complex closes β before releasing it around DNA. Using a fluorescent ATP hydrolysis assay, we show that there is a burst of ATP hydrolysis before β closing and that β release may be the rate-limiting step in the overall clamp loading reaction. The combined use of these fluorescent assays provides a unique perspective into the E. coli clamp loader by providing a measure of the relative timing of different events in the clamp loading reaction, helping to elucidate the complicated clamp loading mechanism. PMID:23161545

  18. Improving safety and efficiency during emergent central venous catheter placement with a needleless securing clamp.

    Science.gov (United States)

    Silich, Bert; Chrobak, Paul; Siu, Jeffrey; Schlichting, Adam; Patel, Samir; Yang, James

    2013-08-01

    To compare the needleless securing clamp to the traditional suture-secured clamp for central venous catheters. Compare the holding strength of each type of clamps by measuring the amount of kinetic energy absorbed, ask 20 physicians to evaluate the clamp placement using sutures or staples, and summarise the clamps effectiveness and complications in 10 patients. Compared to sutured clamp, the needleless clamp was more secure. The needleless clamp was also significantly better with regard to ease of use, safety, perceived strength (p value clamps. Without incurring complications or increasing risk to patients, the needleless clamp is secure and improves safety and efficiency for physicians.

  19. The ATP Sites of AAA+ Clamp Loaders Work Together as a Switch to Assemble Clamps on DNA*

    Science.gov (United States)

    Marzahn, Melissa R.; Hayner, Jaclyn N.; Finkelstein, Jeff; O'Donnell, Mike; Bloom, Linda B.

    2014-01-01

    Clamp loaders belong to a family of proteins known as ATPases associated with various cellular activities (AAA+). These proteins utilize the energy from ATP binding and hydrolysis to perform cellular functions. The clamp loader is required to load the clamp onto DNA for use by DNA polymerases to increase processivity. ATP binding and hydrolysis are coordinated by several key residues, including a conserved Lys located within the Walker A motif (or P-loop). This residue is required for each subunit to bind ATP. The specific function of each ATP molecule bound to the Saccharomyces cerevisiae clamp loader is unknown. A series of point mutants, each lacking a single Walker A Lys residue, was generated to study the effects of abolishing ATP binding in individual clamp loader subunits. A variety of biochemical assays were used to analyze the function of ATP binding during discrete steps of the clamp loading reaction. All mutants reduced clamp binding/opening to different degrees. Decreased clamp binding activity was generally correlated with decreases in the population of open clamps, suggesting that differences in the binding affinities of Walker A mutants stem from differences in stabilization of proliferating cell nuclear antigen in an open conformation. Walker A mutations had a smaller effect on DNA binding than clamp binding/opening. Our data do not support a model in which each ATP site functions independently to regulate a different step in the clamp loading cycle to coordinate these steps. Instead, the ATP sites work in unison to promote conformational changes in the clamp loader that drive clamp loading. PMID:24436332

  20. The ATP sites of AAA+ clamp loaders work together as a switch to assemble clamps on DNA.

    Science.gov (United States)

    Marzahn, Melissa R; Hayner, Jaclyn N; Finkelstein, Jeff; O'Donnell, Mike; Bloom, Linda B

    2014-02-28

    Clamp loaders belong to a family of proteins known as ATPases associated with various cellular activities (AAA+). These proteins utilize the energy from ATP binding and hydrolysis to perform cellular functions. The clamp loader is required to load the clamp onto DNA for use by DNA polymerases to increase processivity. ATP binding and hydrolysis are coordinated by several key residues, including a conserved Lys located within the Walker A motif (or P-loop). This residue is required for each subunit to bind ATP. The specific function of each ATP molecule bound to the Saccharomyces cerevisiae clamp loader is unknown. A series of point mutants, each lacking a single Walker A Lys residue, was generated to study the effects of abolishing ATP binding in individual clamp loader subunits. A variety of biochemical assays were used to analyze the function of ATP binding during discrete steps of the clamp loading reaction. All mutants reduced clamp binding/opening to different degrees. Decreased clamp binding activity was generally correlated with decreases in the population of open clamps, suggesting that differences in the binding affinities of Walker A mutants stem from differences in stabilization of proliferating cell nuclear antigen in an open conformation. Walker A mutations had a smaller effect on DNA binding than clamp binding/opening. Our data do not support a model in which each ATP site functions independently to regulate a different step in the clamp loading cycle to coordinate these steps. Instead, the ATP sites work in unison to promote conformational changes in the clamp loader that drive clamp loading.

  1. Medial Clamp Tine Positioning Affects Ankle Syndesmosis Malreduction.

    Science.gov (United States)

    Cosgrove, Christopher T; Putnam, Sara M; Cherney, Steven M; Ricci, William M; Spraggs-Hughes, Amanda; McAndrew, Christopher M; Gardner, Michael J

    2017-08-01

    To determine whether the position of the medial clamp tine during syndesmotic reduction affected reduction accuracy. Prospective cohort. Urban Level 1 trauma center. Seventy-two patients with operatively treated syndesmotic injuries. Patients underwent operative fixation of their ankle syndesmotic injuries using reduction forceps. The position of the medial clamp tine was then recorded with intraoperative fluoroscopy. Malreduction rates were then assessed with bilateral ankle computerized tomography. Fibular position within the incisura was measured with respect to the uninjured side to determine whether a malreduction had occurred. Malreductions were then analyzed for associations with injury pattern, patient demographics, and the location of the medial clamp tine. A statistically significant association was found between medial clamp position and sagittal plane syndesmosis malreduction. In reference to anterior fibular translation, there was a 0% malreduction rate in the 18 patients where the clamp tine was placed in the anterior third, a 19.4% malreduction rate in the middle third, and 60% malreduction rate in the posterior third (P = 0.006). In reference to posterior fibular translation, there was a 11.1% malreduction when clamp placement was in the anterior third, a 16.1% malreduction rate in the middle third, and 60% malreduction rate in the posterior third (P = 0.062). There were no significant associations between medial clamp position and coronal plane malreductions (overcompression or undercompression) (P = 1). When using reduction forceps for syndesmotic reduction, the position of the medial clamp tine can be highly variable. The angle created with off-axis syndesmotic clamping is likely a major culprit in iatrogenic malreduction. Sagittal plane malreduction appears to be highly sensitive to clamp obliquity, which is directly related to the medial clamp tine placement. Based on these data, we recommend placing the medial clamp tine in the anterior third

  2. Delayed versus Immediate Cord Clamping in Preterm Infants.

    Science.gov (United States)

    Tarnow-Mordi, William; Morris, Jonathan; Kirby, Adrienne; Robledo, Kristy; Askie, Lisa; Brown, Rebecca; Evans, Nicholas; Finlayson, Sarah; Fogarty, Michael; Gebski, Val; Ghadge, Alpana; Hague, Wendy; Isaacs, David; Jeffery, Michelle; Keech, Anthony; Kluckow, Martin; Popat, Himanshu; Sebastian, Lucille; Aagaard, Kjersti; Belfort, Michael; Pammi, Mohan; Abdel-Latif, Mohamed; Reynolds, Graham; Ariff, Shabina; Sheikh, Lumaan; Chen, Yan; Colditz, Paul; Liley, Helen; Pritchard, Margo; de Luca, Daniele; de Waal, Koert; Forder, Peta; Duley, Lelia; El-Naggar, Walid; Gill, Andrew; Newnham, John; Simmer, Karen; Groom, Katie; Weston, Philip; Gullam, Joanna; Patel, Harshad; Koh, Guan; Lui, Kei; Marlow, Neil; Morris, Scott; Sehgal, Arvind; Wallace, Euan; Soll, Roger; Young, Leslie; Sweet, David; Walker, Susan; Watkins, Andrew; Wright, Ian; Osborn, David; Simes, John

    2017-12-21

    The preferred timing of umbilical-cord clamping in preterm infants is unclear. We randomly assigned fetuses from women who were expected to deliver before 30 weeks of gestation to either immediate clamping of the umbilical cord (≤10 seconds after delivery) or delayed clamping (≥60 seconds after delivery). The primary composite outcome was death or major morbidity (defined as severe brain injury on postnatal ultrasonography, severe retinopathy of prematurity, necrotizing enterocolitis, or late-onset sepsis) by 36 weeks of postmenstrual age. Analyses were performed on an intention-to-treat basis, accounting for multiple births. Of 1634 fetuses that underwent randomization, 1566 were born alive before 30 weeks of gestation; of these, 782 were assigned to immediate cord clamping and 784 to delayed cord clamping. The median time between delivery and cord clamping was 5 seconds and 60 seconds in the respective groups. Complete data on the primary outcome were available for 1497 infants (95.6%). There was no significant difference in the incidence of the primary outcome between infants assigned to delayed clamping (37.0%) and those assigned to immediate clamping (37.2%) (relative risk, 1.00; 95% confidence interval, 0.88 to 1.13; P=0.96). The mortality was 6.4% in the delayed-clamping group and 9.0% in the immediate-clamping group (P=0.03 in unadjusted analyses; P=0.39 after post hoc adjustment for multiple secondary outcomes). There were no significant differences between the two groups in the incidences of chronic lung disease or other major morbidities. Among preterm infants, delayed cord clamping did not result in a lower incidence of the combined outcome of death or major morbidity at 36 weeks of gestation than immediate cord clamping. (Funded by the Australian National Health and Medical Research Council [NHMRC] and the NHMRC Clinical Trials Centre; APTS Australian and New Zealand Clinical Trials Registry number, ACTRN12610000633088 .).

  3. Delayed cord clamping and cord gas analysis at birth.

    Science.gov (United States)

    Xodo, Serena; Xodo, Luigi; Berghella, Vincenzo

    2018-01-01

    Delayed cord clamping for at least 60 s in both term and preterm babies is a major recent change in clinical care. Delayed cord clamping has several effects on other possible interventions. One of these is the effect of delayed cord clamping on umbilical artery gas analysis. When indicated, umbilical artery gas analysis can safely be done either with early cord clamping or, probably most of the times it is necessary, during delayed cord clamping with the cord still unclamped. Paired blood samples (one from the umbilical artery and one from the umbilical vein) can be taken from the pulsating and unclamped cord, immediately after birth, during delayed cord clamping, without any effect on either the accuracy of umbilical artery gas analysis or the transfusion of blood through delayed cord clamping. Umbilical artery gas analysis should instead not be done after delayed cord clamping, since delayed cord clamping alters several acid-based parameters and lactate values. © 2017 Nordic Federation of Societies of Obstetrics and Gynecology.

  4. The Use of Gastrostomy Tube for the Long-Term Remission of Hyperinsulinemic Hypoglycemia After Roux-en-y Gastric Bypass

    DEFF Research Database (Denmark)

    Craig, CM; Lamendola, C; Holst, Jens Juul

    2015-01-01

    Objective: Hyperinsulinemic hypoglycemia is an increasingly reported complication of Roux-en-Y gastric bypass surgery (RYGB), for which there is currently no acceptable treatment. We present a case of the reversal of severe hyperinsulinemic hypoglycemia through gastrostomy tube (GT) feeding...... to the remnant stomach and uniquely report the durable resolution of neuroglycopenic symptoms 3 years after GT placement. Methods: The case subject underwent standardized postprandial measurement of plasma glucose, insulin, glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic peptide (GIP......), and glucagon concentrations after oral or GT administration of a standardized liquid meal. Results: Hypersecretion of insulin, GLP-1, and glucagon elicited by oral administration of the liquid meal were reversed with GT feeding. GIP was not secreted in excess of normal after the oral meal. Conclusion...

  5. Off-Clamp versus On-Clamp Robotic Partial Nephrectomy: A Multicenter Match-Paired Case-Control Study.

    Science.gov (United States)

    Peyronnet, Benoit; Khene, Zine-Eddine; Pradère, Benjamin; Seisen, Thomas; Verhoest, Grégory; Masson-Lecomte, Alexandra; Grassano, Yohann; Roumiguié, Mathieu; Beauval, Jean-Baptiste; Baumert, Hervé; Droupy, Stéphane; Doumerc, Nicolas; Bernhard, Jean-Christophe; Vaessen, Christophe; Bruyère, Franck; de la Taille, Alexandre; Rouprêt, Morgan; Bensalah, Karim

    2017-01-01

    The aim of this study was to compare the outcomes of on-clamp and off-clamp robotic partial nephrectomy (RPN). The charts of all patients who underwent an RPN at 8 institutions between 2010 and 2014 were retrospectively reviewed. The patients who underwent an off-clamp RPN were matched to on-clamp RPN in a 1-4 fashion according to the following variables: RENAL score, tumor size and surgeon's experience. Pre-, intra-, and postoperative data were compared between both groups. Among 525 RPN, 26 were performed off-clamp (5%). They were matched to 104 on-clamp RPN. The complications rate (15.5 vs. 7.7%, p = 0.53), major complications rate (4.9 vs. 3.9%; p = 0.82), and transfusions rate (0 vs. 4.9%; p = 0.58) did not differ significantly between the clamped and unclamped groups. Conversely, estimated blood loss was higher in the off-clamp group (266.4 vs. 284.6 mL, p = 0.048) and so was the rate of conversion to radical nephrectomy (0 vs. 7.7%, p = 0.04). Postoperative preservation of renal function was comparable in both groups. Off-clamp RPN is feasible for a small subgroup of renal tumors without increased risk of postoperative complications but at the cost of higher estimated blood loss and increased risk of conversion to radical nephrectomy. © 2017 S. Karger AG, Basel.

  6. Comparison of off-clamp partial nephrectomy and on-clamp partial nephrectomy: a systematic review and meta-analysis.

    Science.gov (United States)

    Trehan, Abhishek

    2014-01-01

    To compare peri- and postoperative variables, surgical complications, oncological outcomes and renal outcomes of off-clamp partial nephrectomy (PN) and on-clamp PN. A systematic search of the electronic databases, including MEDLINE, Embase and Cochrane Library, was performed. The pooled estimates of tumour size, operative time, estimated blood loss, length of stay, overall complications, transfusion rates, urinary leaks, positive surgical margins and eGFR were calculated. 14 studies were included. There was no significant difference between off-clamp PN and on-clamp PN in terms of tumour size, operative time, estimated blood loss, length of stay, overall complications, transfusion rates, urinary leaks, and positive surgical margins. However, a non-statistically significant trend towards increased blood loss (p = 0.12) and transfusion rates (p = 0.07) in those undergoing off-clamp PN was noted. Off-clamp PN was associated with a significantly lower reduction in eGFR than on-clamp PN (standardised weighted mean difference 0.27, 95% CI 0.14, 0.40, p clamp PN may be associated with improved long-term renal outcomes when compared to on-clamp PN with no difference in in peri- and postoperative variables, surgical complications and oncological outcomes. However, the meta-analysis was limited by the design of the underlying studies, and hence further work is necessary. 2014 S. Karger AG, Basel.

  7. Dental dam clamp adaptation method on carved gypsum cast

    Science.gov (United States)

    Cazacu, NCE

    2014-01-01

    Rationale. Dental Dam is the safest and most efficient isolation technique in endodontics and restorative dentistry, but it also used in esthetics, orthodontics, prosthetics, pedodontics and periodontology (for teeth immobilization). While in most cases the standard clamps are efficient, in some clinical situations clamp adaptation is mandatory in order to assure a tight contact on the tooth. Objective. The purpose of this study is to list the elements of the clamp, which should be modified in order to assure a secure constriction of the clamp on the anchor tooth, by using the carved gypsum cast method. Methods and Results. 100 patients were examined, diagnosed and treated for various diagnoses like simple decay, gangrene, chronic apical periodontitis, and endodontic retreatments. The clamps used in this study were produced by Hu-Friedy, Hygenic, KKD, SDI, Hager & Werken. In 10 cases, the anchor tooth did not provide enough stability to the standard clamp – as provided by the producer. Therefore, we have done some adjustments to some of the elements of the clamp: the arch, the wings, the plateau, the active area, and the contact points. In 6 cases, major clamp adaptations on carved gypsum cast were imperative. Discussion. The classic clamps cannot provide a grip to be enough in all the clinical cases due to the huge variety and position and implantation of the anchor teeth. Therefore, in such situations, the clamps should be adapted in order to provide stability and assure the safe isolation during the treatment. The modified clamps will be useful in similar cases, so they must be kept. PMID:25713609

  8. Increased skeletal muscle capillarization enhances insulin sensitivity

    DEFF Research Database (Denmark)

    Åkerström, Thorbjörn; Laub, Lasse; Vedel, Kenneth

    2014-01-01

    . Skeletal muscle specific angiogenesis was induced by adding the α1-adrenergic receptor antagonist Prazosin to the drinking water of Sprague Dawley rats (n=33) while 34 rats served as controls. Insulin sensitivity was measured ≥40 h after termination of the 3-week Prazosin treatment, which ensured...... that Prazosin was cleared from the blood stream. Whole-body insulin sensitivity was measured in conscious, unrestrained rats by hyperinsulinemic euglycemic clamp. Tissue specific insulin sensitivity was assessed by administration of 2-deoxy-[(3)H]-Glucose during the plateau phase of the clamp. Whole-body...

  9. Applications of alcohol clamping in early drug development

    NARCIS (Netherlands)

    Zoethout, Remco Wiebe Martijn

    2012-01-01

    This thesis describes the development of a novel alcohol clamp, a new method to obtain stable plasma levels of alcohol and its application in CNS-research. The method might have several advantages that were explored in subsequent studies described in this thesis. The stability of the alcohol clamp

  10. The problem of isotropic rectangular plate with four clamped edges

    Indian Academy of Sciences (India)

    This report discusses in exact solution of the governing equation of an isotropic rectangular plate with four clamped edges. A numerical method for clamped isotropic rectangular plate under distributed loads and an exact solution of the governing equation in terms of trigonometric and hyperbolic function are given. Finally ...

  11. New active load voltage clamp for HF-link converters

    DEFF Research Database (Denmark)

    Ljusev, Petar; Andersen, Michael Andreas E.

    2005-01-01

    This paper proposes a new active clamp for HF-link converters, which features very high efficiency by returning the clamped energy back to the primary side through a small auxiliary converter. It also increases the reliability of HF-link converters by providing an alternative load current path...

  12. Laparoscopic off-clamp partial nephrectomy using soft coagulation.

    Science.gov (United States)

    Hongo, Fumiya; Kawauchi, Akihiro; Ueda, Takashi; Fujihara-Iwata, Atsuko; Nakamura, Terukazu; Naya, Yoshio; Kamoi, Kazumi; Okihara, Koji; Miki, Tsuneharu

    2015-08-01

    To assess the effectiveness of soft coagulation in off-clamp laparoscopic partial nephrectomy. A total of 32 patients with renal tumors underwent laparoscopic partial nephrectomy with off-clamp using soft coagulation between May 2012 and September 2013. Tumor resection was carried out using a combination of bipolar forceps and a ball electrode using the soft coagulation system without hilar clamping. The outcomes of these patients were compared with those of 30 patients treated with hilar clamping. This off-clamp procedure was successfully completed in 31 cases. No significant differences were observed in the mean age (60 vs 61 years), sex (male/female; 25/7 vs 20/10), mean RENAL nephrometry score (5.7 vs 5.8), mean body mass index (24.4 vs 23) or tumor size (15 mm vs 16 mm) between the two groups. No significant differences were noted in positive surgical margins (0 vs 0) or blood loss (104 vs 115 cc) as well. In contrast, a significant difference was noted in the total operative time (278 vs 238 min). Serum creatinine percentage changes at 3 months were 6.4 versus 7.3% in the off-clamp and hilum-clamp groups, respectively, which were not significantly different. Off-clamp laparoscopic partial nephrectomy can be safely carried out by using a soft coagulation technique. © 2015 The Japanese Urological Association.

  13. Advanced patch-clamp techniques and single-channel analysis

    NARCIS (Netherlands)

    Biskup, B; Elzenga, JTM; Homann, U; Thiel, G; Wissing, F; Maathuis, FJM

    Much of our knowledge of ion-transport mechanisms in plant cell membranes comes from experiments using voltage-clamp. This technique allows the measurement of ionic currents across the membrane, whilst the voltage is held under experimental control. The patch-clamp technique was developed to study

  14. Studying mechanosensitive ion channels with an automated patch clamp

    NARCIS (Netherlands)

    Barthmes, Maria; Jose, Mac Donald F; Birkner, Jan Peter; Brüggemann, Andrea; Wahl-Schott, Christian; Kocer, Armagan

    Patch clamp electrophysiology is the main technique to study mechanosensitive ion channels (MSCs), however, conventional patch clamping is laborious and success and output depends on the skills of the operator. Even though automated patch systems solve these problems for other ion channels, they

  15. Off-clamp robotic partial nephrectomy: Technique and outcome.

    Science.gov (United States)

    Lamoshi, Abdulraouf Y; Salkini, Mohamad W

    2015-01-01

    Robotic partial nephrectomy (RPN) is a technically challenging procedure. Advanced skills are needed to accomplish tumor resection, hemostasis, and renorrhaphy within short ischemia time in RPN. Off-clamp RPN with zero ischemia may decrease the risk of ischemic reperfusion injury to the kidney. However, the off-clamp technique has been associated with an increased risk of blood loss. The purpose of this study was to evaluate the outcome of our modified off-clamp technique utilized in certain RPN cases. A total of 81 patients underwent RPN between September 2009 and July 2013 for renal masses. We studied a subgroup of patients who underwent off-clamp RPN with zero ischemia time. The off-clamp technique was utilized for exophytic, nonhilar tumors that have a base of 2 cm or less. We developed a novel technique to avoid ischemia reperfusion renal injury while minimizing blood loss in certain cases of RPN. Of the 81 cases of RPN, we reviewed and adopted the off-clamp technique in 34 patients (41.98%). Utilizing off-clamp RPN resulted in an average blood loss of 96.29 ml and 1.56 days (range: 1-3 days) of hospital stay and minimal change in serum creatinine. Off-clamp RPN is safe and feasible approach to excise certain kidney tumors. It carries the benefits of RPN and prevents ischemia reperfusion renal injury.

  16. Circumferential hoof clamp method of lameness induction in the horse.

    Science.gov (United States)

    Swaab, M E; Mendez-Angulo, J L; Groschen, D M; Ernst, N S; Brown, M P; Trumble, T N

    2015-07-01

    A circumferential hoof clamp method to induce controlled and reversible lameness in the forelimbs of eight horses was assessed. Peak vertical forces and vertical impulses were recorded using a force plate to verify induced lameness. Video recordings were used by blinded observers to determine subjective lameness using a 0-5 scale and any residual lameness following clamp loosening. Tightening of clamps resulted in consistent, visible lameness in the selected limbs in all horses. Lameness was confirmed by significant decreases from baseline in the peak vertical force (P <0.01). Lameness was also confirmed subjectively by elevated median scores (0 at baseline and 2 during lameness). Lameness was not immediately reversible after clamp loosening (median score 1.5), but horses were not obviously lame after clamp removal and were no different from initial baseline (median score 0.5) approximately 3 days later. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Functional analyses of novel mutations in the sulfonylurea receptor 1 associated with persistent hyperinsulinemic hypoglycemia of infancy.

    Science.gov (United States)

    Shyng, S L; Ferrigni, T; Shepard, J B; Nestorowicz, A; Glaser, B; Permutt, M A; Nichols, C G

    1998-07-01

    The ATP-sensitive potassium channel, K(ATP) channel, a functional complex of the sulfonylurea receptor 1, SUR1, and an inward rectifier potassium channel subunit, Kir6.2, regulates insulin secretion in the pancreas. Mutations in both the Kir6.2 and SUR1 genes are associated with persistent hyperinsulinemic hypoglycemia of infancy (PHHI), a disorder of pancreatic beta-cell function characterized by excess insulin secretion and hypoglycemia. We have studied the functional properties of novel SUR1 mutations identified in PHHI patients, including H125Q, N188S, F591L, T1139M, R1215Q, G1382S, and R1394H. R1394H and deltaF1388 SUR1, a previously identified PHHI mutation, resulted in no functional channels when coexpressed with Kir6.2 in COS cells, while H125Q, N188S, F591L, T1139M, R1215Q, and G1382S SUR1 generated functional channels in the absence of ATP. With the exception of N188S and H125Q, all mutants had reduced response to stimulation by MgADP. These results indicate that lack of, or reduction of, K(ATP) channel sensitivity to MgADP is a common molecular defect associated with the disease. The mutant channels also showed varied response to activation by the potassium channel opener diazoxide. Because these mutations are distributed throughout the molecule, our data have new implications for structure-function relationships of the K(ATP) channel, suggesting that structural elements in SUR1 outside of the two nucleotide-binding folds are also important in regulating channel activity.

  18. Randomized clinical trial of BiClamp forceps versus clamp-crushing technique in open liver resection.

    Science.gov (United States)

    Chen, Jiang Ming; Geng, Wei; Zhang, Song; Liu, Fu Bao; Zhao, Hong Chuan; Zhao, Yi Jun; Wang, Guo Bin; Xie, Sheng Xue; Geng, Xiao Ping

    2017-03-01

    The aim of this trial was to compare the efficacy and safety of BiClamp forceps with the "gold-standard" clamp-crushing technique for open liver resection. From October 2014 to May 2016, 86 consecutive patients scheduled to undergo hepatic resection were randomized to a BiClamp forceps group (n = 43) or to a clamp-crushing technique group (n = 43). Background characteristics of the two groups were closely matched. There were no significant differences between the BiClamp forceps group and clamp-crushing group in total intraoperative blood loss (339.81 ± 257.20 ml vs. 376.73 ± 303.67 ml, respectively; P = 0.545) or blood loss per transection area (5.35 ± 3.27 ml/cm2 vs. 5.44 ± 3.02 ml/cm2 , respectively; P = 0.609). Liver transection speed, the need of blood transfusion, morbidity, length of postoperative hospital stay, total hospitalization cost and liver function recovery were similar in the two groups. Multivariate logistic regression analysis identified major hepatectomy, multiple resections and liver transection time ≥30 min as significantly unfavorable factors for decreased intraoperative blood loss. Liver parenchymal transection with BiClamp forceps is as safe and feasible as the gold-standard clamp-crushing technique. © 2017 Japanese Society of Hepato-Biliary-Pancreatic Surgery.

  19. Is "Delayed Umbilical Cord Clamping" Beneficial for Premature Newborns?

    Directory of Open Access Journals (Sweden)

    Amir-Mohammad Armanian

    2017-05-01

    Full Text Available Background: The appropriate moment for clamping the umbilical cord is controversial. Immediate cord clamping (ICC is an item of active management of the third stage of labor (AMTSL. Unclamped umbilical cord may cause inconvenience in preterm neonates because they commonly need some levels of emergent services. Some studies revealed delayed cord clamping (DCC of preterm neonates results in better health conditions like lower rates of respiratory distress syndrome (RDS, less morbidities in labor room and lower risk of postpartum hemorrhage. The aim of the present study was to determine the effect of delayed umbilical cord clamping on premature neonatal outcomes. Materials and Methods: In this single‑center randomized control trial study, sixty premature neonates (gestational age ≤ 34 weeks were randomly assigned to ICC (cord clamped at 5–10 seconds or DCC (30–45 seconds groups and followed up in neonatal intensive care unit (NICU. Primary outcomes were 1st and 5th minute Apgar score, average of level of hematocrit after birth, intra ventricle hemorrhage and need some levels of resuscitation. Results: Differences in demographic characteristics were not statistically significant. After birth, neonates who had delayed clamping had significantly higher mean hematocrit after at 4-hour of birth (49.58+5.15gr/dl vs. 46.58+5.40gr/dlin DCC vs. ICC groups, respectively (P=0.031. Delayed cord clamping reduced the duration of need to nasal continues positive airway pressure (NCPAP (86.7% and 60.0% in ICC and DCC groups, respectively, P= 0.039. Attractively, the results showed lower incidence of clinical sepsis in delayed cord clamping neonates (53.3% vs. 23.3% in ICC and DCC groups, respectively, P=0.033. Conclusion: Prematurity complications might decrease by delay umbilical cord clamping which improve the hematocrit, duration of need to NCPAP and incidence of clinical sepsis. Furthermore, DCC may have no negative impact on neonatal resuscitation.

  20. Reciprocating clamp apparatus for thermoforming plastic containers

    Energy Technology Data Exchange (ETDEWEB)

    Beck, M.H.; Harry, I.L.; Krishnakumar, S.M.

    1984-03-06

    This relates to the forming of containers and like hollow articles from sheets or webs of thermoplastic material. Two webs or sheets are simultaneously acted upon by way of a forming apparatus which includes a reciprocating clamp first cooperable with one outer platen and then the other in sequence wherein, while a first web or sheet is being formed within a plurality of mold cavities to define a plurality of hollow articles such as containers, the other sheet or web may be stripped from its respective mold set and a new sheet or a new portion of a sheet or web may be advanced into position for molding. The forming apparatus may be constructed in a manner wherein the web portions which are to be formed may be billowed away from the mold cavities as an initial step in the stretching and orientation of the thermoplastic material. The thermoplastic material may be heated to the desired forming temperature using separate sets of rf electrodes so that only those portions of the thermoplastic material which are to be formed need be heated. Two sets of containers of different sizes may be formed from the webs or sheets, and then internested to form double wall containers wherein the walls may be formed of different materials and wherein the outer material may be a barrier material, or wherein the walls of the containers are spaced apart to form an insulated container. This abstract forms no part of the specification of this application and is not to be construed as limiting the claims of the application.

  1. Dynamic Study of a Capacitive MEMS Switch with Double Clamped-Clamped Microbeams

    Directory of Open Access Journals (Sweden)

    Hatem Samaali

    2014-01-01

    Full Text Available We study a capacitive MEMS switch composed of two clamped-clamped exible microbeams. We first develop a mathematical model for the MEMS switch where the upper microbeam represents the ground transmission line and the lower one represents the central transmission line. An electrostatic force is applied between the two microbeams to yield the switch to its ON and OFF states. We derive the equations of motion of the system and associated boundary conditions and solve the static and dynamic problems using the differential quadratic method. We show that using only nine grid points gives relatively accurate results when compared to those obtained using FEM. We also examine the transient behavior of the microswitch and obtain results indicating that subsequent reduction in actuation voltage, switching time, and power consumption are expected along with relatively good RF performances. ANSYS HFSS simulator is used in this paper to extract the RF characteristics of the microswitch. HFSS simulation results show that the insertion loss is as low as −0.31 dB and that the return loss is better than −12.41 dB at 10 GHz in the ON state. At the OFF state, the isolation is lower than −23 dB in the range of 10 to 50 GHz.

  2. Comparison of drain clamp after bilateral total knee arthroplasty.

    Science.gov (United States)

    Madadi, Firooz; Mehrvarz, Amir Sarshekeh; Madadi, Firoozeh; Boreiri, Majid; Abachizadeh, Kambiz; Ershadi, Ali

    2010-12-01

    Suction drains provide an easy and feasible method for controlling hemorrhage after total knee arthroplasty. However, there has been no compromise regarding the optimum clamping time for these drains. We conducted a randomized clinical trial in 50 patients to compare 12-hour drain clamping and continuous drainage after total knee arthroplasty in terms of wound complications, blood loss, and articular arc of motion. To eliminate any other factor except duration of clamping, we chose to compare only knees belonging to a single patient and to restrict the study to those knees undergoing surgery due to osteoarthritis. From a total of 100 knees (50 patients) studied, the 12-hour-clamping method resulted in a significantly smaller amount of postoperative blood loss (p < 0.001). The passive ranges of motion and wound complications were not significantly different between the two groups.

  3. Lotus birth, a holistic approach on physiological cord clamping.

    Science.gov (United States)

    Zinsser, Laura A

    2017-09-04

    The positive effects of delayed cord clamping (DCC) has been extensively researched. DCC means: waiting at least one minute after birth before clamping and cutting the cord or till the pulsation has stopped. With physiological clamping and cutting (PCC) the clamping and cutting can happen at the earliest after the pulsation has stopped. With a Lotus birth, no clamping and cutting of the cord is done. A woman called Clair Lotus Day imitated the holistic approach of PCC from an anthropoid ape in 1974. The chimpanzee did not separate the placenta from the newborn. The aim of this case report is to discuss and learn a different approach in the third stage of labour. Three cases of Lotus birth by human beings were observed. All three women gave birth in an out-of-hospital setting and had ambulant postnatal care. The placenta was washed, salted and herbs were put on 2-3h post partum. The placenta was wrapped in something that absorbs the moisture. The salting was repeated with a degreasing frequency depending on moistness of the placenta. On life day six all three Lotus babies experiences a natural separation of the cord. All three Lotus birth cases were unproblematic, no special incidence occurred. One should differentiate between early cord clamping (ECC), delayed cord clamping (DCC) and physiological cord clamping (PCC). Lotus birth might lead to an optimisation of the bonding and attachment. Research is needed in the areas of both PCC and Lotus birth. Copyright © 2017 Australian College of Midwives. Published by Elsevier Ltd. All rights reserved.

  4. Force-controlled patch clamp of beating cardiac cells.

    Science.gov (United States)

    Ossola, Dario; Amarouch, Mohamed-Yassine; Behr, Pascal; Vörös, János; Abriel, Hugues; Zambelli, Tomaso

    2015-03-11

    From its invention in the 1970s, the patch clamp technique is the gold standard in electrophysiology research and drug screening because it is the only tool enabling accurate investigation of voltage-gated ion channels, which are responsible for action potentials. Because of its key role in drug screening, innovation efforts are being made to reduce its complexity toward more automated systems. While some of these new approaches are being adopted in pharmaceutical companies, conventional patch-clamp remains unmatched in fundamental research due to its versatility. Here, we merged the patch clamp and atomic force microscope (AFM) techniques, thus equipping the patch-clamp with the sensitive AFM force control. This was possible using the FluidFM, a force-controlled nanopipette based on microchanneled AFM cantilevers. First, the compatibility of the system with patch-clamp electronics and its ability to record the activity of voltage-gated ion channels in whole-cell configuration was demonstrated with sodium (NaV1.5) channels. Second, we showed the feasibility of simultaneous recording of membrane current and force development during contraction of isolated cardiomyocytes. Force feedback allowed for a gentle and stable contact between AFM tip and cell membrane enabling serial patch clamping and injection without apparent cell damage.

  5. Analysis to Effect of Non-linear Axial Force on the MEMS Clamped-clamped Switch Beam

    Directory of Open Access Journals (Sweden)

    Cao Tian-Jie

    2014-04-01

    Full Text Available The effect of non-linear axial force, which is caused by both temperature change of environment and the length change of the beam, on the behavior of the switch is quite heavy because of the larger slenderness ratio of the MEMS clamped-clamped switch beams. By using the 1D beam model, the behaviors of electrostatic MEMS clamped-clamped switch beams with both the temperature change and the length change for three different stages were studied. In the paper, the governing equation of the elastic curve of the beam with the effect of non-linear axial force in the beam was given. A three-fold method of bisection was used to solve the unknown axial force in the governing equation, applied voltage and one boundary condition of the beam. On obtaining the axial force, the applied voltage and the boundary condition, the deflections at any cross-section could be obtained easily by using the numerical method to the ordinary differential equation. Incorporating the numerical method to the ordinary differential equation with some optimization method, pull-in voltage and the corresponding deflection could be distinguished and obtained automatically on a computer with the specifically designed program. In examples, the entire mechanical behavior of the MEMS clamped- clamped switch beam over the operational voltage ranges was investigated and the effects of the axial force on pull-in voltage at several temperature changes with different geometry dimensions of the beam were studied.

  6. Analyzing of Nonuniform Hardness for Anchorage Clamp Plate after Heat Treatment

    Science.gov (United States)

    Liu, Zhenmin; Wang, Lijun; Ruan, Shipeng; Wang, Ningtao

    2017-09-01

    20CrMnTi wire rod can be used to manufacture the anchorage clamp plate. In the normal production, the test results show that the hardness of anchorage clamp plate is nonuniform. In this paper, the chemical composition, microstructure, grain size of clamp plate billet and the microstructure, hardness of the finished clamp plate product were studied and analyzed. The results show that improper heat treatment technics causes nonuniform hardness for anchorage clamp plate.

  7. [Advantage of delayed umbilical cord clamping in the newborn infant].

    Science.gov (United States)

    Menget, A; Mougey, C; Thiriez, G; Riethmuller, D

    2013-09-01

    The timing of umbilical cord clamping remains controversial. Although most maternity wards use the early clamping (5-15s), randomized studies and meta-analyses have demonstrated the benefit of delayed clamping for term and preterm newborn infants over the past 10 years. Indeed, placentofetal transfusion of 20-30 ml/kg in 2-3 min improves the iron status of term infants and prevents infant hypochromic anemia. Infant anemia is a public health problem in many developing countries. For preterm newborns, placental transfusion for 45 s or milking the cord for 15 s improves cardiovascular adaptation, with better hemodynamic stability, as well as decreased intraventricular hemorrhages, need for transfusion, and late-onset sepsis. A new look at this symbolic act is needed and professionals need to be persuaded of the importance of the "wait a minute" policy for a better physiological delivery. Copyright © 2013. Published by Elsevier SAS.

  8. New active load voltage clamp for HF-link converters

    Energy Technology Data Exchange (ETDEWEB)

    Ljusev, P.; Andersen, M.A.E.

    2005-07-01

    This paper proposes a new active clamp for HF-link converters, which features very high efficiency by returning the clamped energy back to the primary side through a small auxiliary converter. It also increases the reliability of HF-link converters by providing an alternative load current path during malfunctions of the secondary bidirectional bridge. The feasibility of the approach is shown on audio power amplifier prototype. New integrated magnetics design is presented that incorporates both the main power and auxiliary transformer on the same magnetic core. (au)

  9. Plasma temperature clamping in filamentation laser induced breakdown spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Harilal, Sivanandan S.; Yeak, J.; Phillips, Mark C.

    2015-10-19

    Ultrafast laser filament induced breakdown spectroscopy is a very promising method for remote material detection. We present characteristics of plasmas generated in a metal target by laser filaments in air. Our measurements show that the temperature of the ablation plasma is clamped along the filamentation channel due to intensity clamping in a filament. Nevertheless, significant changes in radiation intensity are noticeable, and this is essentially due to variation in the number density of emitting atoms. The present results also partly explains the reason for the occurrence of atomic plume during fs LIBS in air compared to long-pulse ns LIBS.

  10. Insulin secretion and cellular glucose metabolism after prolonged low-grade intralipid infusion in young men

    DEFF Research Database (Denmark)

    Jensen, Christine B; Storgaard, Heidi; Holst, Jens Juul

    2003-01-01

    not in the nonoxidative) glucose metabolism in young healthy men. Moreover, insulin hypersecretion perfectly countered the free-fatty acid-induced insulin resistance. Future studies are needed to determine the role of a prolonged moderate lipid load in subjects at increased risk of developing diabetes.......We examined the simultaneous effects of a 24-h low-grade Intralipid infusion on peripheral glucose disposal, intracellular glucose partitioning and insulin secretion rates in twenty young men, by 2-step hyperinsulinemic euglycemic clamp [low insulin clamp (LI), 10 mU/m(2) x min; high insulin clamp...... (HI), 40 mU/m(2) x min], 3-(3)H-glucose, indirect calorimetry, and iv glucose tolerance test. Free fatty acid concentrations were similar during basal steady state but 3.7- to 13-fold higher during clamps. P-glucagon increased and the insulin/glucagon ratio decreased at both LI and HI during...

  11. Oral chromium picolinate improves carbohydrate and lipid metabolism and enhances skeletal muscle Glut-4 translocation in obese, hyperinsulinemic (JCR-LA corpulent) rats.

    Science.gov (United States)

    Cefalu, William T; Wang, Zhong Q; Zhang, Xian H; Baldor, Linda C; Russell, James C

    2002-06-01

    Human studies suggest that chromium picolinate (CrPic) decreases insulin levels and improves glucose disposal in obese and type 2 diabetic populations. To evaluate whether CrPic may aid in treatment of the insulin resistance syndrome, we assessed its effects in JCR:LA-corpulent rats, a model of this syndrome. Male lean and obese hyperinsulinemic rats were randomly assigned to receive oral CrPic [80 microg/(kg. d); n = 5 or 6, respectively) in water or to control conditions (water, n = 5). After 3 mo, a 120-min intraperitoneal glucose tolerance test (IPGTT) and a 30-min insulin tolerance test were performed. Obese rats administered CrPic had significantly lower fasting insulin levels (1848 +/- 102 vs. 2688 +/- 234 pmol/L; P < 0.001; mean +/- SEM) and significantly improved glucose disappearance (P < 0.001) compared with obese controls. Glucose and insulin areas under the curve for IPGTT were significantly less for obese CrPic-treated rats than in obese controls (P < 0.001). Obese CrPic-treated rats had lower plasma total cholesterol (3.57 +/- 0.28 vs. 4.11 +/- 0.47 mmol/L, P < 0.05) and higher HDL cholesterol levels (1.92 +/- 0.09 vs. 1.37 +/- 0.36 mmol/L, P < 0.01) than obese controls. CrPic did not alter plasma glucose or cholesterol levels in lean rats. Total skeletal muscle glucose transporter (Glut)-4 did not differ among groups; however, CrPic significantly enhanced membrane-associated Glut-4 in obese rats after insulin stimulation. Thus, CrPic supplementation enhances insulin sensitivity and glucose disappearance, and improves lipids in male obese hyperinsulinemic JCR:LA-corpulent rats.

  12. A new configuration for multilevel converters with diode clamped topology

    DEFF Research Database (Denmark)

    Nami, A.; Zare, F.; Ledwich, G.

    2008-01-01

    points in this type of converter is dc-voltage control. In this paper, a novel multi output dc-dc converter connected to a diode clamped topology is presented. This converter, for a given duty cycles, is able to regulate the capacitor voltage to provide an appropriate input voltage for NPC regardless...

  13. Improved DNA clamps by stacking to adjacent nucleobases

    DEFF Research Database (Denmark)

    Fatthalla, M.I.; Pedersen, Erik Bjerregaard

    2012-01-01

    to the underlying non-planar base triplet in the formed triplex. This type of triplexes has a substantially higher thermal melting temperature which can be further improved by inserting locked nucleic acids (LNAs) in the Hoogsteen part of the clamp. An extremely high sensitivity to mismatches is observed...

  14. Beyond the patch clamp: nanotechnologies for intracellular recording.

    Science.gov (United States)

    Kruskal, Peter B; Jiang, Zhe; Gao, Teng; Lieber, Charles M

    2015-04-08

    The patch clamp is a fundamental tool for neuroscientists, offering insights that have shaped our understanding of the brain. Advances in nanotechnology suggest that the next generation of recording methods is now within reach. We discuss the complexity and future promise of applying nanoscience to neural recording. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. A piezoelectric vibration harvester based on clamped-guided beams

    NARCIS (Netherlands)

    Wang, Z.; Matova, S.; Elfrink, R.; Jambunathan, M.; Nooijer, C. de; Schaijk, R. van; Vullers, R.J.M.

    2012-01-01

    The paper addresses the design, modeling, fabrication and experimental results of a piezoelectric energy harvester based on clamped-guided beams. The design is featured by shorter mass displacement and higher reliability than cantilever beams. Two separate sets of capacitors allow exploiting both

  16. The cardiovascular effects of aortic clamping and unclamping

    African Journals Online (AJOL)

    presence of overt coronary artery disease. 2. The radius of the ventricle influences wall tension. In other words .... follows acute aortic constriction. This recovery of end-diastolic volume and pressure toward control .... concomitant valvular disease is beyond the scope of this manuscript. The circulation proximal to the clamp.

  17. Five-level Z-source diode-clamped inverter

    DEFF Research Database (Denmark)

    Gao, F.; Loh, Poh Chiang; Blaabjerg, Frede

    2010-01-01

    This study proposes a five-level Z-source diode-clamped inverter designed with two intermediate Z-source networks connected between the dc input sources and rear-end inverter circuitry. By partially shorting the Z-source networks, new operating states not previously reported for two-level Z...

  18. Significance of the clamp-connection in the Basidiomycetes

    NARCIS (Netherlands)

    Furtado, João S.

    1966-01-01

    The cytogenetic phenomena affecting clamp-connection formation and the interpretation of this peculiar type of septum in the classification of the higher Basidiomycetes were reviewed from the literature and discussed. The cytogenetic data available are restricted to a small number of basidiomycetous

  19. Technical Note. A Clamped Bar Model for the Sompoton Vibrator

    National Research Council Canada - National Science Library

    Tee Hao Wong; Jedol Dayou; M.C.D. Ngu; Jackson H.W. Chang; Willey Y.H. Liew

    2013-01-01

    .... In this paper, the vibrator is modeled as a clamped bar with a uniformly distributed mass. By means of this model, the fundamental frequency is analyzed with the use of an equivalent single degree of freedom system (SDOF) and exact analysis...

  20. analytical bending solution of all clamped isotropic rectangular plate

    African Journals Online (AJOL)

    HP

    to the governing differential equation of the plate on Winkler foundation to obtain the deflection coefficient, . Numerical example was presented at the end to compare the results obtained by this method and those from earlier studies. The percentage difference obtained for central deflection of all clamped rectangular plate ...

  1. INOVASI SAMBUNGAN MEKANIS MENGGUNAKAN CLAMP BAJA UNTUK TULANGAN BETON

    Directory of Open Access Journals (Sweden)

    Parmo

    2015-12-01

    Full Text Available In the last few years, Indonesian urban growth has tends to be lead to the overground space. This was caused by the significant population increasement the lackness of land availability. The phenomenon is also marked with the increasing number of high-rise buildings and skyscrapers. However, high-rise buildings and skyscrapers have the potential dangers against earthquakes, even more Indonesia lay on an earthquake-prone areas. There are two main things that need to be considered in the planning of earthquake-resistant buildings, which is strength and ductility. Deformation capability and the connection strength between the reinforcement taken into consideration in the design of earthquake-resistant structures. From the steel clamp tensile test results for D13’s mechanical connection reinforcement, it gained 270.69 Mpa for yield strength, 351.45 MPa for ultimate tensile strength and 4757 kg for maximum load with 40% elongation. As for the D16’s mechanical connection reinforcement gained 217.80 MPa for yield strength, 327 605 MPa for ultimate tensile strength and 6717 kg for maximum load with 32% elongation. This study uses two pieces of steel clamp, but in order to obtain a better result then it need to be add more clamps while increasing the steel clamp material quality.

  2. A clamp-like biohybrid catalyst for DNA oxidation

    NARCIS (Netherlands)

    van Dongen, S.F.M.; Clerx, J.; Norgaard, K.; Bloemberg, T.G.; Cornelissen, Jeroen Johannes Lambertus Maria; Trakselis, M.A.; Nelson, S.W.; Benkovic, S.J.; Rowan, A.E.; Nolte, R.J.M.

    2013-01-01

    In processive catalysis, a catalyst binds to a substrate and remains bound as it performs several consecutive reactions, as exemplified by DNA polymerases. Processivity is essential in nature and is often mediated by a clamp-like structure that physically tethers the catalyst to its (polymeric)

  3. The problem of isotropic rectangular plate with four clamped edges

    Indian Academy of Sciences (India)

    ... received considerable attention because of its technical importance. This paper analyses the deflections of an isotropic rectangular clamped thin plates under uniformly distributed loads. A plate is called thin when its thickness is at least one order of magnitude smaller than the span of the plate. The bending and buckling.

  4. The phylogenetics of dynamics in DNA clamp proteins.

    Science.gov (United States)

    Balasubramaniam, Deepa; Komives, Elizabeth A

    2014-04-08

    Using HXMS to measure protein dynamics, Fang and colleagues (in this issue of Structure) probe the dynamics of processivity clamp proteins from all kingdoms of life. The proteins have similar structures but divergent sequences. Their results show conserved dynamics correlating with primary functions and divergent dynamics for divergent functions. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Increased rate of whole body lipolysis before and after 9 days of bed rest in healthy young men born with low birth weight

    DEFF Research Database (Denmark)

    Alibegovic, Amra Ciric; Hojbjerre, Lise; Sonne, Mette

    2010-01-01

    Background: Individuals born with low birth weight (LBW) are at risk of developing type 2 diabetes mellitus (T2D), which may be precipitated by physical inactivity. Methods: 22 LBW and 23 controls were studied before and after bed rest by the hyperinsulinemic euglycemic clamp combined with indirect...... calorimetry, infusion of stable isotope tracers, preceded by an intravenous glucose tolerance test. Results: LBW subjects had similar BMI, but elevated abdominal obesity compared with controls. The basal rate of whole body lipolysis (WBL) was elevated in LBW subjects with and without correction for abdominal...

  6. A two-week reduction of ambulatory activity attenuates peripheral insulin sensitivity

    DEFF Research Database (Denmark)

    Krogh-Madsen, Rikke; Thyfault, John P; Broholm, Christa

    2009-01-01

    men decreased their daily activity level from a mean of 10,501 (+/- 808) to 1,344 (+/- 33) steps/day for 2 weeks. Hyperinsulinemic-euglycemic clamps with stable isotopes and muscle biopsies, maximal oxygen consumption (VO2max) tests, and blood samples were performed pre and post intervention.A reduced......US adults take between ~2,000 to ~12,000 steps per day, a wide range of ambulatory activity, that at the low range could increase risk for developing chronic metabolic diseases. Dramatic reductions in physical activity induce insulin resistance; however it is uncertain if and how low ambulatory...

  7. A 2-wk reduction of ambulatory activity attenuates peripheral insulin sensitivity

    DEFF Research Database (Denmark)

    Krogh-Madsen, Rikke; Thyfault, John P; Broholm, Christa

    2010-01-01

    healthy young men decreased their daily activity level from a mean of 10,501+/-808 to 1,344+/-33 steps/day for 2 wk. Hyperinsulinemic-euglycemic clamps with stable isotopes and muscle biopsies, maximal oxygen consumption (VO2 max) tests, and blood samples were performed pre- and postintervention......US adults take between approximately 2,000 and approximately 12,000 steps per day, a wide range of ambulatory activity that at the low range could increase risk for developing chronic metabolic diseases. Dramatic reductions in physical activity induce insulin resistance; however, it is uncertain...

  8. Early enhancements of hepatic and later of peripheral insulin sensitivity combined with increased postprandial insulin secretion contribute to improved glycemic control after Roux-en-Y gastric bypass

    DEFF Research Database (Denmark)

    Bojsen-Møller, Kirstine N; Dirksen, Carsten; Jørgensen, Nils Bruun

    2014-01-01

    after RYGB. Participants were included after a preoperative diet induced total weight loss of -9.2±1.2%. Hepatic and peripheral insulin sensitivity were assessed using the hyperinsulinemic euglycemic clamp combined with glucose tracer technique and beta-cell function evaluated in response...... to an intravenous glucose-glucagon challenge as well as an oral glucose load. Already within 1 week, RYGB reduced basal glucose production, improved basal hepatic insulin sensitivity and increased insulin clearance highlighting the liver as an important organ responsible for the early effects on glucose metabolism...... in response to oral glucose, underscoring the importance of the changed gut anatomy....

  9. Single-nucleotide polymorphism rs7754840 of CDKAL1 is associated with impaired insulin secretion in nondiabetic offspring of type 2 diabetic subjects and in a large sample of men with normal glucose tolerance

    DEFF Research Database (Denmark)

    Stancáková, Alena; Pihlajamäki, Jussi; Kuusisto, Johanna

    2008-01-01

    -sectional study including subjects from five white populations in Europe (Denmark, Finland, Germany, Italy, and Sweden). Study 2 is an ongoing prospective study of Finnish men. PARTICIPANTS: In study 1, 846 nondiabetic offspring of type 2 diabetic patients (age 40 +/- 10 yr; body mass index 26.7 +/- 5.0 kg/m(2......)) participated. In study 2, subjects included 3900 middle-aged men (533 type 2 diabetic and 3367 nondiabetic subjects). Interventions: Interventions included iv glucose-tolerance test (IVGTT), oral glucose-tolerance test (OGTT), and euglycemic-hyperinsulinemic clamp in study 1 and OGTT in study 2. MAIN OUTCOME...

  10. Short-term weight loss attenuates local tissue inflammation and improves insulin sensitivity without affecting adipose inflammation in obese mice

    OpenAIRE

    Jung, Dae Young; Ko, Hwi Jin; Lichtman, Eben I.; Lee, Eunjung; Lawton, Elizabeth; Ong, Helena; Yu, Kristine; Azuma, Yoshihiro; Friedline, Randall H; Lee, Ki Won; Kim, Jason K.

    2013-01-01

    Obesity is a major cause of insulin resistance, and weight loss is shown to improve glucose homeostasis. But the underlying mechanism and the role of inflammation remain unclear. Male C57BL/6 mice were fed a high-fat diet (HFD) for 12 wk. After HFD, weight loss was induced by changing to a low-fat diet (LFD) or exercise with continuous HFD. The weight loss effects on energy balance and insulin sensitivity were determined using metabolic cages and hyperinsulinemic euglycemic clamps in awake mi...

  11. Retinol-binding protein 4 in twins: regulatory mechanisms and impact of circulating and tissue expression levels on insulin secretion and action

    DEFF Research Database (Denmark)

    Ribel-Madsen, Rasmus; Friedrichsen, Martin; Vaag, Allan

    2009-01-01

    OBJECTIVE: Retinol-binding protein (RBP) 4 is an adipokine of which plasma levels are elevated in obesity and type 2 diabetes. The aims of the study were to identify determinants of plasma RBP4 and RBP4 mRNA expression in subcutaneous adipose tissue (SAT) and skeletal muscle and to investigate...... twins. Peripheral and hepatic insulin sensitivity was assessed by a euglycemic-hyperinsulinemic clamp, and beta-cell function was estimated from an intravenous glucose tolerance test. RESULTS: The influence of environmental versus genetic factors in the regulation of plasma RBP4 increased with age...

  12. A Low-Glycemic Diet Lifestyle Intervention Improves Fat Utilization during Exercise in Older Obese Humans

    DEFF Research Database (Denmark)

    Solomon, Thomas; Haus, Jacob M; Cook, Marc A

    2013-01-01

    Objective: To determine the influence of dietary glycemic index on exercise training-induced adaptations in substrate oxidation in obesity. Design and Methods: Twenty older, obese individuals undertook 3 months of fully supervised aerobic exercise and were randomized to low- (LoGIX) or high......-glycemic (HiGIX) diets. Changes in indirect calorimetry (VO2 ; VCO2 ) were assessed at rest, during a hyperinsulinemic-euglycemic clamp, and during submaximal exercise (walking: 65% VO2 max, 200 kcal energy expenditure). Intramyocellular lipid (IMCL) was measured by (1) H-magnetic resonance spectroscopy...

  13. Dietary fat drives whole-body insulin resistance and promotes intestinal inflammation independent of body weight gain

    DEFF Research Database (Denmark)

    Jensen, Benjamin Anderschou Holbech; Nielsen, Thomas Svava; Fritzen, Andreas Mæchel

    2016-01-01

    BACKGROUND: The obesogenic potential of high-fat diets (HFD) in rodents is attenuated when the protein:carbohydrate ratio is increased. However, it is not known if intake of an HFD irrespective of the protein:carbohydrate ratio and in the absence of weight gain, affects glucose homeostasis...... no differences were observed during insulin tolerance tests. The hyperinsulinemic euglycemic clamp revealed tissue-specific effects on glucose homeostasis in all HFD-fed groups. HFD-fed mice exhibited decreased insulin-stimulated glucose uptake in white but not in brown adipose tissue, and sustained endogenous...

  14. Altered PI3-kinase/Akt signalling in skeletal muscle of young men with low birth weight

    DEFF Research Database (Denmark)

    Jensen, C.B.; Martin-Gronert, M.S.; Storgaard, H.

    2008-01-01

    regulatory subunit and p110beta catalytic subunit, PKCzeta and GLUT4 in the fasting state. The aim of this study was to determine whether insulin activation of the PI3K/Akt and MAPK signalling pathways is altered in skeletal muscle of young adult men with LBW. METHODS: Vastus lateralis muscle biopsies were...... obtained from 20 healthy 19-yr old men with BWage (LBW) and 20 normal birth weight controls (NBW), matched for physical fitness and whole-body glucose disposal, prior to (fasting state) and following a 4-hr hyperinsulinemic euglycemic clamp (insulin stimulated state...... stimulated levels of IRS1 (psignalling...

  15. Complement factors C4 and C3 are down regulated in response to short term overfeeding in healthy young men

    DEFF Research Database (Denmark)

    Foghmar, Caroline; Brøns, Charlotte; Pilely, Katrine

    2017-01-01

    -days HFO by a hyperinsulinemic-euglycemic-clamp. Circulating C4, C3, ficolins, mannose-binding-lectin, complement activation products C3bc, terminal complement complex (TCC) and complement activation capacity were determined using turbidimetry and ELISA. HFO induced peripheral insulin resistance in LBW...... individuals only, while both groups had the same degree of hepatic insulin resistance after HFO. Viewing all individuals circulating levels of C4, C3, C3bc, TCC and complement activation capacity decreased paradoxically along the development of insulin resistance after HFO (P = 0.0015, P

  16. Preliminary report: genetic variation within the GPBAR1 gene is not associated with metabolic traits in white subjects at an increased risk for type 2 diabetes mellitus

    DEFF Research Database (Denmark)

    Müssig, Karsten; Staiger, Harald; Machicao, Fausto

    2009-01-01

    -cell dysfunction. We genotyped 1576 subjects (1043 women, 533 men) for the single nucleotide polymorphism rs3731859 in the GPBAR1 gene. All subjects underwent an oral glucose tolerance test; a subset additionally had a hyperinsulinemic-euglycemic clamp. Regional fat distribution, ectopic hepatic...... and intramyocellular lipids were determined by magnetic resonance techniques. Peak aerobic capacity, a surrogate parameter for oxidative capacity of skeletal muscle, was measured by an incremental exercise test on a motorized treadmill. Total GLP-1 and gastric inhibitory peptide levels were determined...

  17. To clamp or not to clamp? Bladder management by suprapubic catheterization in patients with neurogenic bladder dysfunction.

    Science.gov (United States)

    Pannek, Jürgen; Göcking, Konrad; Bersch, Ulf

    2010-10-01

    In a small subset of patients with neurogenic lower urinary tract dysfunction, insertion of suprapubic catheters (SPC) cannot be avoided. If SPC has to be utilized, catheter clamping and anticholinergic medication are often recommended, but evidence supporting this view is scarce. We determined the influence of anticholinergic medication and catheter clamping on urodynamic parameters and the status of the urinary tract in patients with chronic suprapubic catheterization. In a retrospective study, the results of urodynamic testing, sonographic evaluations, and urinalyses of 85 patients with chronic (>1 year) suprapubic catheterization due to neurogenic bladder dysfunction were analyzed. The 51 male and 34 female patients (mean age 55 years) were managed with an SPC for 65.3 ± 48.0 months. Forty patients had an SPC for more than 60 months. Comparing the results before SPC insertion with the last follow-up examination, no significant differences in detrusor compliance and maximum detrusor pressure were detected, whereas bladder capacity significantly decreased. In three patients, alterations of the upper urinary tract were found. The results were not significantly different between the patients using anticholinergic medication and/or catheter clamping and those who did not. According to our study, routine use of anticholinergic medication and clamping of catheter does not seem to be necessary to preserve detrusor compliance and renal function in patients with SPC and neurogenic bladder dysfunction.

  18. Impact of Early Release of the IVC Clamp from Different IVC Clamping Maneuvers on Changes in Hemodynamic Parameters in Living Donor Liver Transplantation.

    Science.gov (United States)

    Shih, Tsung-Hsiao; Huang, Chia-Jung; Chen, Chao-Long; Cheng, Kwok-Wai; Wu, Shao-Chun; Yang, Sheng-Chun; Juang, Sin-Ei; Lee, Ying-En; Jawan, Bruno; Wang, Chih-Hsien

    2015-09-06

    BACKGROUND The aim of this study was to evaluate the impact of different methods of inferior vena cava (IVC) clamping and release of the cross clamp on hemodynamic parameters of recipients during living donor liver transplantation. MATERIAL AND METHODS Ninety-six adult living donor liver transplantation patients were divided into 3 groups according to cross-clamp of the IVC for all the hepatic vein and portal vein reconstruction (G1), cross-clamp of the IVC only for hepatic vein reconstruction (G2), and side-clamp of the IVC for hepatic vein reconstruction (G3). In G2 and G2, the reconstructed hepatic vein was clamped instead of the IVC for portal vein reconstruction. The hemodynamic parameters among groups were compared by 1-way ANOVA and the complications in each group were compared using the Kruskal-Wallis test. RESULTS Changes in percentage of MAP and CO in G3 were significantly less than that of G1 and G2 for hepatic vein reconstruction. Hemodynamic parameters of G2 and G3 normalized to pre-clamped values during portal vein reconstruction, while the hemodynamics of G1 remained unstable. CONCLUSIONS Hemodynamic changes were less pronounced in LT with side-clamp of the inferior cava vein versus total cross-clamp. Early release of the IVC clamp minimized the hemodynamic changes. There were no differences in terms of outcome (morbidity and mortality).

  19. Patch-Clamp Recording from Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes: Improving Action Potential Characteristics through Dynamic Clamp.

    Science.gov (United States)

    Verkerk, Arie O; Veerman, Christiaan C; Zegers, Jan G; Mengarelli, Isabella; Bezzina, Connie R; Wilders, Ronald

    2017-08-30

    Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold great promise for studying inherited cardiac arrhythmias and developing drug therapies to treat such arrhythmias. Unfortunately, until now, action potential (AP) measurements in hiPSC-CMs have been hampered by the virtual absence of the inward rectifier potassium current (IK1) in hiPSC-CMs, resulting in spontaneous activity and altered function of various depolarising and repolarising membrane currents. We assessed whether AP measurements in "ventricular-like" and "atrial-like" hiPSC-CMs could be improved through a simple, highly reproducible dynamic clamp approach to provide these cells with a substantial IK1 (computed in real time according to the actual membrane potential and injected through the patch-clamp pipette). APs were measured at 1 Hz using perforated patch-clamp methodology, both in control cells and in cells treated with all-trans retinoic acid (RA) during the differentiation process to increase the number of cells with atrial-like APs. RA-treated hiPSC-CMs displayed shorter APs than control hiPSC-CMs and this phenotype became more prominent upon addition of synthetic IK1 through dynamic clamp. Furthermore, the variability of several AP parameters decreased upon IK1 injection. Computer simulations with models of ventricular-like and atrial-like hiPSC-CMs demonstrated the importance of selecting an appropriate synthetic IK1. In conclusion, the dynamic clamp-based approach of IK1 injection has broad applicability for detailed AP measurements in hiPSC-CMs.

  20. Patch-Clamp Recording from Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes: Improving Action Potential Characteristics through Dynamic Clamp

    Science.gov (United States)

    Veerman, Christiaan C.; Zegers, Jan G.; Mengarelli, Isabella; Bezzina, Connie R.

    2017-01-01

    Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold great promise for studying inherited cardiac arrhythmias and developing drug therapies to treat such arrhythmias. Unfortunately, until now, action potential (AP) measurements in hiPSC-CMs have been hampered by the virtual absence of the inward rectifier potassium current (IK1) in hiPSC-CMs, resulting in spontaneous activity and altered function of various depolarising and repolarising membrane currents. We assessed whether AP measurements in “ventricular-like” and “atrial-like” hiPSC-CMs could be improved through a simple, highly reproducible dynamic clamp approach to provide these cells with a substantial IK1 (computed in real time according to the actual membrane potential and injected through the patch-clamp pipette). APs were measured at 1 Hz using perforated patch-clamp methodology, both in control cells and in cells treated with all-trans retinoic acid (RA) during the differentiation process to increase the number of cells with atrial-like APs. RA-treated hiPSC-CMs displayed shorter APs than control hiPSC-CMs and this phenotype became more prominent upon addition of synthetic IK1 through dynamic clamp. Furthermore, the variability of several AP parameters decreased upon IK1 injection. Computer simulations with models of ventricular-like and atrial-like hiPSC-CMs demonstrated the importance of selecting an appropriate synthetic IK1. In conclusion, the dynamic clamp-based approach of IK1 injection has broad applicability for detailed AP measurements in hiPSC-CMs. PMID:28867785

  1. Review: The lord of the rings: Structure and mechanism of the sliding clamp loader.

    Science.gov (United States)

    Kelch, Brian A

    2016-08-01

    Sliding clamps are ring-shaped polymerase processivity factors that act as master regulators of cellular replication by coordinating multiple functions on DNA to ensure faithful transmission of genetic and epigenetic information. Dedicated AAA+ ATPase machines called clamp loaders actively place clamps on DNA, thereby governing clamp function by controlling when and where clamps are used. Clamp loaders are also important model systems for understanding the basic principles of AAA+ mechanism and function. After nearly 30 years of study, the ATP-dependent mechanism of opening and loading of clamps is now becoming clear. Here I review the structural and mechanistic aspects of the clamp loading process, as well as comment on questions that will be addressed by future studies. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 532-546, 2016. © 2016 Wiley Periodicals, Inc.

  2. Four- year thyroidectomy procedures: Ligasure Vs. clamp & tie

    Directory of Open Access Journals (Sweden)

    Pazoki AR

    2010-01-01

    Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: The electrothermal bipolar vessel sealing system is an adjunct to the hemostatic technique, which has been successfully used in many abdominal operation, recently made available to thyroid surgery. The purpose of this was compare the outcomes of new hemostatic technique with traditional method of clamp and tie."n"nMethods: In this retrospective cross-sectional study has been done in surgery's wards of Milad & Rasul- Akram hospitals in Tehran, Iran. During three- year period between 2005-2008 all patients with different kind of pathology that need operation were assessed. Time duration of surgery and hospital stay and post surgery complication in two group, ligasure (n=45 and clamp & tie (n=145 were compared with each others."n"nResults: No difference was seen between two groups in time duration of surgery. Significant difference was seen in hospital stay and complications of surgery. Time of operation in Ligasure group was 132/22 minute and in clamp and tie group was 130/10 minute. There was found significant difference in duration of hospital stay and postoperative complication. Hypocalcemia were seen in two and 24 cases in Ligasure and Clamp & Tie group, respectively (p<0.05. Hospital stay was 2

  3. Open partial nephrectomy with selective renal parenchymal control: a new reliable clamp.

    Science.gov (United States)

    Huyghe, Eric; Nohra, Joe; Leobon, Bertrand; El Khoury, Elias; Khedis, Mehdi; Soulié, Michel; Plante, Pierre

    2006-09-01

    We describe our experience of open partial nephrectomy with a parenchymal clamp, the Reni-Clamp, in 30 patients from January 2002 to May 2005. The mean operative and clamping time was 150 and 27 minutes, respectively, and the blood loss was 150 mL. The Reni-Clamp enabled us to perform partial nephrectomy safely in all cases of polar or external edge renal tumor.

  4. Hepatectomy for spontaneous rupture of hepatocellular carcinoma without portal triad clamping.

    Science.gov (United States)

    Wang, Zhi-Ming; Lu, Xin-Sheng; Qiu, Fu

    2017-10-01

    Hepatectomy without portal triad clamping may decrease the incidence of liver injury; however, the effects of hepatectomy without portal triad clamping in the treatment of spontaneous rupture of hepatocellular carcinoma (SRHCC) remain unclear. The aims of the present study were to evaluate the therapeutic value of hepatectomy without portal triad clamping in the treatment of patients with SRHCC. The present study retrospectively reviewed patients with SRHCC who received hepatectomy without portal triad clamping (non-clamping group) and the therapeutic efficacy was compared with that of 20 patients with SRHCC undergoing the same surgery in the presence of portal triad clamping (clamping group). Following hepatectomy, the non-clamping group exhibited a significantly lower incidence of acute liver failure compared with the clamping group (P0.05). At 1 week and 2 weeks after surgery, the non-clamping group exhibited significantly lower alanine aminotransferase, aspartate aminotransferase and total bilirubin serum levels compared with the clamping group (all Pportal triad clamping may decrease the incidence of liver injury and liver failure in patients with SRHCC, suggesting that it may be a safe and effective therapeutic strategy.

  5. MEMS-based clamp with a passive hold function for precision position retaining of micromanipulators

    NARCIS (Netherlands)

    Brouwer, Dannis Michel; de Jong, B.R.; de Boer, Meint J.; Jansen, Henricus V.; van Dijk, Johannes A.G.M.; van Dijk, Johannes; Krijnen, Gijsbertus J.M.; Soemers, Herman

    2009-01-01

    In this paper the design, modeling and fabrication of a precision MEMS-based clamp with arelatively large clamping force are presented. The purpose of the clamp is to mechanically fixa six-degree-of-freedom (DOF) MEMS-based sample manipulator (Brouwer et al J. Int. Soc. Precis. Eng. Nanotechnol.

  6. File list: Oth.ALL.20.Clamp.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.20.Clamp.AllCell dm3 TFs and others Clamp All cell types SRX159172,SRX15917...0,SRX159174,SRX159166,SRX159173 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/assembled/Oth.ALL.20.Clamp.AllCell.bed ...

  7. File list: Oth.CeL.05.Clamp.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.CeL.05.Clamp.AllCell dm3 TFs and others Clamp Cell line SRX159172,SRX159170,SRX...159174,SRX159166,SRX159173 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/assembled/Oth.CeL.05.Clamp.AllCell.bed ...

  8. File list: Oth.CeL.20.Clamp.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.CeL.20.Clamp.AllCell dm3 TFs and others Clamp Cell line SRX159172,SRX159170,SRX...159174,SRX159166,SRX159173 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/assembled/Oth.CeL.20.Clamp.AllCell.bed ...

  9. File list: Oth.ALL.05.Clamp.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.05.Clamp.AllCell dm3 TFs and others Clamp All cell types SRX159172,SRX15917...0,SRX159174,SRX159166,SRX159173 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/assembled/Oth.ALL.05.Clamp.AllCell.bed ...

  10. File list: Oth.ALL.50.Clamp.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.50.Clamp.AllCell dm3 TFs and others Clamp All cell types SRX159172,SRX15917...0,SRX159174,SRX159166,SRX159173 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/assembled/Oth.ALL.50.Clamp.AllCell.bed ...

  11. File list: Oth.CeL.50.Clamp.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.CeL.50.Clamp.AllCell dm3 TFs and others Clamp Cell line SRX159172,SRX159170,SRX...159174,SRX159166,SRX159173 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/assembled/Oth.CeL.50.Clamp.AllCell.bed ...

  12. File list: Oth.ALL.10.Clamp.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.ALL.10.Clamp.AllCell dm3 TFs and others Clamp All cell types SRX159170,SRX15917...2,SRX159174,SRX159166,SRX159173 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/assembled/Oth.ALL.10.Clamp.AllCell.bed ...

  13. File list: Oth.CeL.10.Clamp.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.CeL.10.Clamp.AllCell dm3 TFs and others Clamp Cell line SRX159170,SRX159172,SRX...159174,SRX159166,SRX159173 http://dbarchive.biosciencedbc.jp/kyushu-u/dm3/assembled/Oth.CeL.10.Clamp.AllCell.bed ...

  14. Gigaseal formation in patch clamping with applications of nanotechnology

    CERN Document Server

    Malboubi, Majid

    2014-01-01

    This book presents an investigation of gigaseal formation using micro/nanotechnology. The aims of the book are twofold. First, it explains the mechanisms of gigaseal formation using the latest discoveries. Second, it provides practical techniques for frequent formation of high resistance seals. The formation of a high-resistance electrical seal, also known as a gigaseal, between a cell membrane and a glass micropipette tip is essential in patch-clamp experiments. Even though four decades have passed since the introduction of the patch-clamping technique by Neher and Sakmann, gigaseal formation remains an obstacle in developing the high-throughput ion channel screening systems required by the pharmaceutical industry. Here the authors share their latest methods for achieving gigaseal formation and describe techniques that are highly desirable at both research and industrial levels. Nanotechnology has been found to be a powerful tool for studying and modifying glass micropipettes and in tackling the problem of g...

  15. Patch Clamp Experiments under Conditions of Variable Graviy

    Science.gov (United States)

    Kohn, F. P. M.; Meissner, K.

    2013-02-01

    The cellular membrane is an intrinsic part of any cell. It has a complex composition of lipid molecules and proteins. The membrane is, among others, involved in excitation and signal transduction. Ion channels, as integral membrane proteins, play an important role. For the question of gravity sensitivity of biological systems, especially neuronal cells, ion channels are of high interest. Gravity might directly interact with the ion channel protein or it might change the thermodynamic membrane parameters, influencing the incorporated proteins indirectly. Detailed information about the effects of gravity on the function of single ion-channels can up to now only be acquired by electrophysiological approaches like the patch clamp technique. Today this technique is the preferentially used technique for single ion-channel studies. Consequently, experiments have been developed in recent years to investigate the interaction of gravity with single ion channel molecules utilizing the patch-clamp technology on different macro- and micro-gravity platforms.

  16. A High-Efficiency Diode-Clamped Linear Amplifier

    Science.gov (United States)

    Fujita, Hideaki

    This paper proposes a new power converter without any switching operation, which works as a linear amplifier. The main circuit of the proposed converter consists of series-connected MOSFETs, series-multi dc power supplies and clamping diodes. The circuit configuration is similar to a diode-clamped multi-level inverter, except for using complementary power devices, which are n- and p-channel MOSFETs. One of the series-connected MOSFETs operates in an active state just like a linear amplifier, while the other MOSFETs operate in on or off states like an inverter circuit. As a result, the proposed converter achieves an acceptable efficiency as high as 90% without any ripples nor harmonics caused by switching operation. Experimental results demonstrate that the proposed converter has capability of driving a 2.2-kW three-phase induction motor.

  17. [Time of cord clamping and neonatal complications, a prospective study].

    Science.gov (United States)

    Rincón, D; Foguet, A; Rojas, M; Segarra, E; Sacristán, E; Teixidor, R; Ortega, A

    2014-09-01

    To assess the effects of early or late clamping of the umbilical cord in term newborns, assessing the levels of hemoglobin, hematocrit, and ferritin, and their correlation with some of the complications. A prospective study of healthy newborns at term or born by dystotic or eutocic delivery in our hospital between May 2009 until May 2010. Patients were assigned according to the time of clamping, group 1 (neonatal phototherapy or admission to the Intensive Care Unit and the hospital stay, were evaluated. A total of 242 patients were included: group 1 (g1=80), group 2 (g2=31) y group 3 (g3=131). The background maternal and neonatal characteristics were similar in all sets. The first test showed significant differences in ferritin levels in those infants with delayed clamping (g1: 111 mg/dl, g2: 125 mg/dl, g3: 173 mg/dl; p<0.01). In the second analysis the values of hemoglobin (g1: 17.3 g/dl, g2: 18.9 g/dl, g3: 19.2 g/dl; p<0.01), hematocrit (g1: 53.4%, g2: 58%, g3: 59%; p<0.01) and ferritin (g1: 254 mg/dl, g2: 254.7 mg/dl, g3: 313 mg/dl; p = 0.008) were statistically higher in this group. As regards complications, a significant increase was observed in the number of cases of polycythemia symptoms in group 3. The late cord clamping is associated with an increase in hematocrit, hemoglobin and ferritin at 48 hours of life, as well as an increased risk of polycythemia present with symptoms. Copyright © 2013 Asociación Española de Pediatría. Published by Elsevier Espana. All rights reserved.

  18. Voltage Balancing Control of Diode-Clamped Multilevel Inverter

    Directory of Open Access Journals (Sweden)

    ŞCHIOP Adrian

    2013-10-01

    Full Text Available In this paper is developed a control scheme for mono-phase diode clamped inverter to achieve balancing voltages on inverter capacitors. First, it develops a control scheme without taking into account the need to balance voltage on two capacitors. It examines the effects on the output voltage inverter, and then it realizes two control schemes that will balance the voltages. The simulations of control schemes were performed in OrCAD Pspice.

  19. The Human Cytomegalovirus UL44 C Clamp Wraps around DNA

    Science.gov (United States)

    Komazin-Meredith, Gloria; Petrella, Robert J.; Santos, Webster L.; Filman, David J.; Hogle, James M.; Verdine, Gregory L.; Karplus, Martin; Coen, Donald M.

    2010-01-01

    Summary Processivity factors tether the catalytic subunits of DNA polymerases to DNA so that continuous synthesis of long DNA strands is possible. The human cytomegalovirus DNA polymerase subunit UL44 forms a C clamp-shaped dimer intermediate in structure between monomeric herpes simplex virus UL42, which binds DNA directly via a basic surface, and the trimeric sliding clamp PCNA, which encircles DNA. To investigate how UL44 interacts with DNA, calculations were performed in which a 12 bp DNA oligonucleotide was docked to UL44. The calculations suggested that UL44 encircles DNA, which interacts with basic residues both within the cavity of the C clamp and in flexible loops of UL44 that complete the “circle.” The results of mutational and crosslinking studies were consistent with this model. Thus, UL44 is a “hybrid” of UL42 and PCNA: its structure is intermediate between the two and its mode of interaction with DNA has elements of both. PMID:18682223

  20. Investigation of the Dynamics of a Clamped-Clamped Microbeam Near the Third Mode Using a Partial Electrode

    KAUST Repository

    Masri, Karim M.

    2014-08-17

    We present an investigation of the dynamics of a clamped-clamped microbeam excited electrostatically near its third mode. To maximize the response at the third mode, a partial electrode configuration is utilized. A multi-mode Galerkin method is used to develop a reduced order model (ROM) of the beam. A shooting method to find the periodic motion is utilized to generate frequency response curves. The curves show hardenining behavior and dynamic pull-in. We show that the dynamic amplitude of the partial configuration is higher than that of a full electrode configuration. These results are promising for the use of higher-order modes for mass detection and for ultra sensitive resonant sensors.

  1. Microsurgical Hepatic Artery Reconstruction Using Ikuta A-II Double Clamp.

    Science.gov (United States)

    Okochi, Masayuki; Okochi, Hiromi; Sakaba, Takao; Momiyama, Masanori; Ueda, Kazuki

    2017-05-01

    We performed hepatic artery (HA) reconstruction on 24 patients between January 2010 and October 2016. Six of 24 patients used an Ikuta type A-II vascular clamp (A-II group). The mean age was 38.0 years (range, 1-61 years). There was no blood leakage at the anastomosed site in any of the patients. No patients required an additional vascular clamp, and none developed HA thrombosis. Eighteen of 24 patients used a conventional vascular clamp. The mean age was 36.1 years (range, 1-65 years; conventional group). Sixteen of 18 patients required an additional vascular clamp due to blood leakage from the HA. There was no significant difference between the 2 groups in mean age or diameter of the recipient HA. However, there was a significant difference in the proportion of patients who required an additional vascular clamp (n clamp is an effective vascular clamp for reconstruction of the HA in living donor liver transplantation.

  2. Dynamic clamp as a tool to study the functional effects of individual membrane currents.

    Science.gov (United States)

    Berecki, Géza; Verkerk, Arie O; van Ginneken, Antoni C G; Wilders, Ronald

    2014-01-01

    Today, the patch-clamp technique is the main technique in electrophysiology to record action potentials or membrane current from isolated cells, using a patch pipette to gain electrical access to the cell. The common recording modes of the patch-clamp technique are current clamp and voltage clamp. In the current clamp mode, the current injected through the patch pipette is under control while the free-running membrane potential of the cell is recorded. Current clamp allows for measurements of action potentials that may either occur spontaneously or in response to an injected stimulus current. In voltage clamp mode, the membrane potential is held at a set level through a feedback circuit, which allows for the recording of the net membrane current at a given membrane potential.A less common configuration of the patch-clamp technique is the dynamic clamp. In this configuration, a specific non-predetermined membrane current can be added to or removed from the cell while it is in free-running current clamp mode. This current may be computed in real time, based on the recorded action potential of the cell, and injected into the cell. Instead of being computed, this current may also be recorded from a heterologous expression system such as a HEK-293 cell that is voltage-clamped by the free-running action potential of the cell ("dynamic action potential clamp"). Thus, one may directly test the effects of an additional or mutated membrane current, a synaptic current or a gap junctional current on the action potential of a patch-clamped cell. In the present chapter, we describe the dynamic clamp on the basis of its application in cardiac cellular electrophysiology.

  3. M. tuberculosis Sliding β-Clamp Does Not Interact Directly with the NAD+ -Dependent DNA Ligase

    Science.gov (United States)

    Kukshal, Vandna; Khanam, Taran; Chopra, Deepti; Singh, Nidhi; Sanyal, Sabyasachi; Ramachandran, Ravishankar

    2012-01-01

    The sliding β-clamp, an important component of the DNA replication and repair machinery, is drawing increasing attention as a therapeutic target. We report the crystal structure of the M. tuberculosis β-clamp (Mtbβ-clamp) to 3.0 Å resolution. The protein crystallized in the space group C2221 with cell-dimensions a = 72.7, b = 234.9 & c = 125.1 Å respectively. Mtbβ-clamp is a dimer, and exhibits head-to-tail association similar to other bacterial clamps. Each monomer folds into three domains with similar structures respectively and associates with its dimeric partner through 6 salt-bridges and about 21 polar interactions. Affinity experiments involving a blunt DNA duplex, primed-DNA and nicked DNA respectively show that Mtbβ-clamp binds specifically to primed DNA about 1.8 times stronger compared to the other two substrates and with an apparent Kd of 300 nM. In bacteria like E. coli, the β-clamp is known to interact with subunits of the clamp loader, NAD+ -dependent DNA ligase (LigA) and other partners. We tested the interactions of the Mtbβ-clamp with MtbLigA and the γ-clamp loader subunit through radioactive gel shift assays, size exclusion chromatography, yeast-two hybrid experiments and also functionally. Intriguingly while Mtbβ-clamp interacts in vitro with the γ-clamp loader, it does not interact with MtbLigA unlike in bacteria like E. coli where it does. Modeling studies involving earlier peptide complexes reveal that the peptide-binding site is largely conserved despite lower sequence identity between bacterial clamps. Overall the results suggest that other as-yet-unidentified factors may mediate interactions between the clamp, LigA and DNA in mycobacteria. PMID:22545130

  4. Insulin secretion and cellular glucose metabolism after prolonged low-grade intralipid infusion in young men

    DEFF Research Database (Denmark)

    Jensen, Christine B; Storgaard, Heidi; Holst, Jens Juul

    2003-01-01

    We examined the simultaneous effects of a 24-h low-grade Intralipid infusion on peripheral glucose disposal, intracellular glucose partitioning and insulin secretion rates in twenty young men, by 2-step hyperinsulinemic euglycemic clamp [low insulin clamp (LI), 10 mU/m(2) x min; high insulin clamp...... Intralipid infusion. At LI, glucose oxidation decreased by 10%, whereas glucose disposal, glycolytic flux, glucose storage, and glucose production were not significantly altered. At HI, glucose disposal, and glucose oxidation decreased by 12% and 24%, respectively, during Intralipid infusion. Glycolytic flux......, glucose storage, and glucose production were unchanged. Insulin secretion rates increased in response to Intralipid infusion, but disposition indices (DI = insulin action.insulin secretion) were unchanged. In conclusion, a 24-h low-grade Intralipid infusion caused insulin resistance in the oxidative (but...

  5. Analysis of motion during the breast clamping phase of mammography.

    Science.gov (United States)

    Ma, Wang Kei; McEntee, Mark F; Mercer, Claire; Kelly, Judith; Millington, Sara; Hogg, Peter

    2016-01-01

    To measure paddle motion during the clamping phase of a breast phantom for a range of machine/paddle combinations. A deformable breast phantom was used to simulate a female breast. 12 mammography machines from three manufacturers with 22 flexible and 20 fixed paddles were evaluated. Vertical motion at the paddle was measured using two calibrated linear potentiometers. For each paddle, the motion in millimetres was recorded every 0.5 s for 40 s, while the phantom was compressed with 80 N. Independent t-tests were used to determine differences in paddle motion between flexible and fixed, small and large, GE Senographe Essential (General Electric Medical Systems, Milwaukee, WI) and Hologic Selenia Dimensions paddles (Hologic, Bedford, MA). Paddle tilt in the medial-lateral plane for each machine/paddle combination was calculated. All machine/paddle combinations demonstrate highest levels of motion during the first 10 s of the clamping phase. The least motion is 0.17 ± 0.05 mm/10 s (n = 20) and the most motion is 0.51 ± 0.15 mm/10 s (n = 80). There is a statistical difference in paddle motion between fixed and flexible (p motion and tilting, and the extent varied with machine and paddle sizes and types. This research suggests that image blurring will likely be clinically insignificant 4 s or more after the clamping phase commences.

  6. Normal-Pressure Tests of Circular Plates with Clamped Edges

    Science.gov (United States)

    Mcpherson, Albert E; Ramberg, Walter; Levy, Samuel

    1942-01-01

    A fixture is described for making normal-pressure tests of flat plates 5 inches in diameter in which particular care was taken to obtain rigid clamping at the edges. Results are given for 19 plates, ranging in thickness form 0.015 to 0.072 inch. The center deflections and the extreme-fiber stresses at low pressures were found to agree with theoretical values; the center deflections at high pressures were 4 to 12 percent greater than the theoretical values. Empirical curves are derived of the pressure for the beginning of the permanent set as a function of the dimensions of the plate and the tensile properties of the material.

  7. Electrostatic Interactions at the Dimer Interface Stabilize the E. coli β Sliding Clamp.

    Science.gov (United States)

    Purohit, Anirban; England, Jennifer K; Douma, Lauren G; Tondnevis, Farzaneh; Bloom, Linda B; Levitus, Marcia

    2017-08-22

    Sliding clamps are ring-shaped oligomeric proteins that encircle DNA and associate with DNA polymerases for processive DNA replication. The dimeric Escherichia coli β-clamp is closed in solution but must adopt an open conformation to be assembled onto DNA by a clamp loader. To determine what factors contribute to the stability of the dimer interfaces in the closed conformation and how clamp dynamics contribute to formation of the open conformation, we identified conditions that destabilized the dimer and measured the effects of these conditions on clamp dynamics. We characterized the role of electrostatic interactions in stabilizing the β-clamp interface. Increasing salt concentration results in decreased dimer stability and faster subunit dissociation kinetics. The equilibrium dissociation constant of the dimeric clamp varies with salt concentration as predicted by simple charge-screening models, indicating that charged amino acids contribute to the remarkable stability of the interface at physiological salt concentrations. Mutation of a charged residue at the interface (Arg-103) weakens the interface significantly, whereas effects are negligible when a hydrophilic (Ser-109) or a hydrophobic (Ile-305) amino acid is mutated instead. It has been suggested that clamp opening by the clamp loader takes advantage of spontaneous opening-closing fluctuations at the clamp's interface, but our time-resolved fluorescence and fluorescence correlation experiments rule out conformational fluctuations that lead to a significant fraction of open states. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  8. Randomised trial of cord clamping and initial stabilisation at very preterm birth.

    Science.gov (United States)

    Duley, Lelia; Dorling, Jon; Pushpa-Rajah, Angela; Oddie, Sam J; Yoxall, Charles William; Schoonakker, Bernard; Bradshaw, Lucy; Mitchell, Eleanor J; Fawke, Joe Anthony

    2018-01-01

    For very preterm births, to compare alternative policies for umbilical cord clamping and immediate neonatal care. Parallel group randomised (1:1) trial, using sealed opaque numbered envelopes. Eight UK tertiary maternity units. 261 women expected to have a live birth before 32 weeks, and their 276 babies. Cord clamping after at least 2 min and immediate neonatal care with cord intact, or clamping within 20 s and immediate neonatal care after clamping. Intraventricular haemorrhage (IVH), death before discharge. 132 women (137 babies) were allocated clamping ≥2 min and neonatal care cord intact, and 129 (139) clamping ≤20 s and neonatal care after clamping; six mother-infant dyads were excluded (2, 4) as birth was after 35+6 weeks, one withdrew (death data only available) (0, 1). Median gestation was 28.9 weeks for those allocated clamping ≥2 min, and 29.2 for those allocated clamping ≤20 s. Median time to clamping was 120 and 11 s, respectively. 7 of 135 infants (5.2%) allocated clamping ≥2 min died and 15 of 135 (11.1%) allocated clamping ≤20 s; risk difference (RD) -5.9% (95% CI -12.4% to 0.6%). Of live births, 43 of 134 (32%) had IVH vs 47 of 132 (36%), respectively; RD -3.5% (-14.9% to 7.8%). There were no clear differences in other outcomes for infants or mothers. This is promising evidence that clamping after at least 2 min and immediate neonatal care with cord intact at very preterm birth may improve outcome; a large trial is urgently needed. ISRCTN 21456601. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  9. The human dynamic clamp as a paradigm for social interaction.

    Science.gov (United States)

    Dumas, Guillaume; de Guzman, Gonzalo C; Tognoli, Emmanuelle; Kelso, J A Scott

    2014-09-02

    Social neuroscience has called for new experimental paradigms aimed toward real-time interactions. A distinctive feature of interactions is mutual information exchange: One member of a pair changes in response to the other while simultaneously producing actions that alter the other. Combining mathematical and neurophysiological methods, we introduce a paradigm called the human dynamic clamp (HDC), to directly manipulate the interaction or coupling between a human and a surrogate constructed to behave like a human. Inspired by the dynamic clamp used so productively in cellular neuroscience, the HDC allows a person to interact in real time with a virtual partner itself driven by well-established models of coordination dynamics. People coordinate hand movements with the visually observed movements of a virtual hand, the parameters of which depend on input from the subject's own movements. We demonstrate that HDC can be extended to cover a broad repertoire of human behavior, including rhythmic and discrete movements, adaptation to changes of pacing, and behavioral skill learning as specified by a virtual "teacher." We propose HDC as a general paradigm, best implemented when empirically verified theoretical or mathematical models have been developed in a particular scientific field. The HDC paradigm is powerful because it provides an opportunity to explore parameter ranges and perturbations that are not easily accessible in ordinary human interactions. The HDC not only enables to test the veracity of theoretical models, it also illuminates features that are not always apparent in real-time human social interactions and the brain correlates thereof.

  10. A reduction clamp for an aiming component in associated acetabular fractures.

    Science.gov (United States)

    Wang, Zhang-Fu; Hong, Zheng-Hua; Wang, Mei-Zhen; Ruan, Jian-Wei; Wang, Wei; Pan, Wei-Bo

    2015-01-01

    The treatment of acetabular fractures is complex and requires specialized equipment. However, all currently available instruments have some disadvantages. A new reduction clamp that can firmly enable reduction and not hinder subsequent fixation procedures for some special fracture types is needed. In this study, we introduce a new acetabular clamp and its preliminary clinical application in three T-shaped acetabular fractures. This new clamp can successfully pull the posterior column back to the anterior column and firmly maintain the reduction. This clamp's aiming plate can facilitate the insertion of long lag screws. The clamp is also easy to assemble and use. This reduction clamp is a useful instrument that can facilitate open reduction and internal fixation of acetabular fractures.

  11. Renal function is the same 6 months after robot-assisted partial nephrectomy regardless of clamp technique: analysis of outcomes for off-clamp, selective arterial clamp and main artery clamp techniques, with a minimum follow-up of 1 year.

    Science.gov (United States)

    Komninos, Christos; Shin, Tae Young; Tuliao, Patrick; Han, Woong Kyu; Chung, Byung Ha; Choi, Young Deuk; Rha, Koon Ho

    2015-06-01

    To compare the renal functional outcomes, with >1 year of follow-up, of patients who underwent robot-assisted partial nephrectomy (RAPN) performed with different clamping techniques. The peri-operative data of patients undergoing RAPN performed with different clamping techniques were retrospectively analysed (group 1: off-clamp, n = 23; group 2: selective clamp, n = 25; group 3: main artery clamp, n = 114). The main outcome measures were postoperative serum creatinine level, estimated glomerular filtration rate (eGFR) and percentage change in eGFR, the data for which were collected at periodic intervals during the first 12 months and annually thereafter, in addition to late eGFR value. Only patients with >1 year of follow-up were included in the analysis. The baseline characteristics of groups 2 and 3 were similar, while patients in group 1 had smaller sized tumours and lower tumour complexity. The median follow-up periods were 45 (group 1), 20 (group 2) and 47 (group 3) months. The median clamping times were 24.8 min in the main artery clamp and 18 min in the selective artery clamp groups. Group 2 had greater median blood loss volume (100 vs 500 vs 200 mL for groups 1, 2 and 3, respectively; P clamp procedure and tumour complexity score were predictive of normal renal function 7 days after surgery, while only baseline eGFR and age could predict it 1 year postoperatively. Off-clamp and selective artery clamp techniques result in superior short-term renal functional outcomes compared with the main artery clamp approach; however, after the 6th postoperative month, there were no significant differences regarding the functional outcome among the above surgical techniques, as long as the warm ischaemia time was 20-30 min. © 2014 The Authors BJU International © 2014 BJU International.

  12. Microchip amplifier for in vitro, in vivo, and automated whole cell patch-clamp recording.

    Science.gov (United States)

    Harrison, Reid R; Kolb, Ilya; Kodandaramaiah, Suhasa B; Chubykin, Alexander A; Yang, Aimei; Bear, Mark F; Boyden, Edward S; Forest, Craig R

    2015-02-15

    Patch clamping is a gold-standard electrophysiology technique that has the temporal resolution and signal-to-noise ratio capable of reporting single ion channel currents, as well as electrical activity of excitable single cells. Despite its usefulness and decades of development, the amplifiers required for patch clamping are expensive and bulky. This has limited the scalability and throughput of patch clamping for single-ion channel and single-cell analyses. In this work, we have developed a custom patch-clamp amplifier microchip that can be fabricated using standard commercial silicon processes capable of performing both voltage- and current-clamp measurements. A key innovation is the use of nonlinear feedback elements in the voltage-clamp amplifier circuit to convert measured currents into logarithmically encoded voltages, thereby eliminating the need for large high-valued resistors, a factor that has limited previous attempts at integration. Benchtop characterization of the chip shows low levels of current noise [1.1 pA root mean square (rms) over 5 kHz] during voltage-clamp measurements and low levels of voltage noise (8.2 μV rms over 10 kHz) during current-clamp measurements. We demonstrate the ability of the chip to perform both current- and voltage-clamp measurement in vitro in HEK293FT cells and cultured neurons. We also demonstrate its ability to perform in vivo recordings as part of a robotic patch-clamping system. The performance of the patch-clamp amplifier microchip compares favorably with much larger commercial instrumentation, enabling benchtop commoditization, miniaturization, and scalable patch-clamp instrumentation. Copyright © 2015 the American Physiological Society.

  13. Crystal structure of a DNA polymerase sliding clamp from a Gram-positive bacterium

    Directory of Open Access Journals (Sweden)

    O'Donnell Mike

    2006-01-01

    Full Text Available Abstract Background Sliding DNA clamps are processivity factors that are required for efficient DNA replication. DNA polymerases maintain proximity to nucleic acid templates by interacting with sliding clamps that encircle DNA and thereby link the polymerase enzyme to the DNA substrate. Although the structures of sliding clamps from Gram-negative bacteria (E. coli, eukaryotes, archaea, and T4-like bacteriophages are well-known, the structure of a sliding clamp from Gram-positive bacteria has not been reported previously. Results We have determined the crystal structure of the dimeric β subunit of the DNA polymerase III holoenzyme of Streptococcus pyogenes. The sliding clamp from this Gram-positive organism forms a ring-shaped dimeric assembly that is similar in overall structure to that of the sliding clamps from Gram-negative bacteria, bacteriophage T4, eukaryotes and archaea. The dimer has overall dimensions of ~90 Å × ~70 Å × ~25 Å with a central chamber that is large enough to accommodate duplex DNA. In comparison to the circular shape of other assemblies, the S. pyogenes clamp adopts a more elliptical structure. Conclusion The sequences of sliding clamps from S. pyogenes and E. coli are only 23% identical, making the generation of structural models for the S. pyogenes clamp difficult in the absence of direct experimental information. Our structure of the S. pyogenes β subunit completes the catalog of clamp structures from all the major sequence grouping of sliding clamps. The more elliptical rather than circular structure of the S. pyogenes clamp implies that the topological nature of encircling DNA, rather than a precise geometric shape, is the most conserved aspect for this family of proteins.

  14. Stepwise unfolding of titin under force-clamp atomic force microscopy

    OpenAIRE

    Andres F. Oberhauser; Hansma, Paul K.; Carrion-Vazquez, Mariano; Fernandez, Julio M.

    2001-01-01

    Here we demonstrate the implementation of a single-molecule force clamp adapted for use with an atomic force microscope. We show that under force-clamp conditions, an engineered titin protein elongates in steps because of the unfolding of its modules and that the waiting times to unfold are exponentially distributed. Force-clamp measurements directly measure the force dependence of the unfolding probability and readily captures the different mechanical stability of the...

  15. Microchip amplifier for in vitro, in vivo, and automated whole cell patch-clamp recording

    Science.gov (United States)

    Kolb, Ilya; Kodandaramaiah, Suhasa B.; Chubykin, Alexander A.; Yang, Aimei; Bear, Mark F.; Boyden, Edward S.; Forest, Craig R.

    2014-01-01

    Patch clamping is a gold-standard electrophysiology technique that has the temporal resolution and signal-to-noise ratio capable of reporting single ion channel currents, as well as electrical activity of excitable single cells. Despite its usefulness and decades of development, the amplifiers required for patch clamping are expensive and bulky. This has limited the scalability and throughput of patch clamping for single-ion channel and single-cell analyses. In this work, we have developed a custom patch-clamp amplifier microchip that can be fabricated using standard commercial silicon processes capable of performing both voltage- and current-clamp measurements. A key innovation is the use of nonlinear feedback elements in the voltage-clamp amplifier circuit to convert measured currents into logarithmically encoded voltages, thereby eliminating the need for large high-valued resistors, a factor that has limited previous attempts at integration. Benchtop characterization of the chip shows low levels of current noise [1.1 pA root mean square (rms) over 5 kHz] during voltage-clamp measurements and low levels of voltage noise (8.2 μV rms over 10 kHz) during current-clamp measurements. We demonstrate the ability of the chip to perform both current- and voltage-clamp measurement in vitro in HEK293FT cells and cultured neurons. We also demonstrate its ability to perform in vivo recordings as part of a robotic patch-clamping system. The performance of the patch-clamp amplifier microchip compares favorably with much larger commercial instrumentation, enabling benchtop commoditization, miniaturization, and scalable patch-clamp instrumentation. PMID:25429119

  16. Microchip amplifier for in vitro, in vivo, and automated whole cell patch-clamp recording

    OpenAIRE

    Harrison, Reid R.; Kolb, Ilya; Kodandaramaiah, Suhasa B.; Chubykin, Alexander A.; Yang, Aimei; Bear, Mark F.; Boyden, Edward S.; Forest, Craig R.

    2014-01-01

    Patch clamping is a gold-standard electrophysiology technique that has the temporal resolution and signal-to-noise ratio capable of reporting single ion channel currents, as well as electrical activity of excitable single cells. Despite its usefulness and decades of development, the amplifiers required for patch clamping are expensive and bulky. This has limited the scalability and throughput of patch clamping for single-ion channel and single-cell analyses. In this work, we have developed a ...

  17. Capparis spinosa L. aqueous extract evokes antidiabetic effect in streptozotocin-induced diabetic mice.

    Science.gov (United States)

    Eddouks, Mohamed; Lemhadri, Ahmed; Hebi, Morad; El Hidani, Ahmed; Zeggwagh, Naoufel Ali; El Bouhali, Bachir; Hajji, Lhoussaine; Burcelin, Remy

    2017-01-01

    As the aqueous extract of Capparis spinosa (CS) possess antidiabetic effect, he present study aims to reveal the possible mechanism of action of CS in diabetic mice. Both single and repeated oral administrations of aqueous extract of CS were performed in multi-low dose streptozotocin-induced (MLDS) diabetic mice. Euglycemic hyperinsulinemic clamp was used in association with the endogenous glucose production (perfusion rate of 3-3H glucose) to evaluate the effect of CS aqueous extract on insulin sensitivity. Our study showed that aqueous extract of CS possess a potent hypoglycaemic activity in MLDS diabetic mice. Furthermore, the analysis perfusion of 3-3H glucose demonstrated the parallel decrease of basal endogenous glucose production (EGP) with the hypoglycaemic activity. EGP was lower in CS-Treated group when compared to the control group (p<0.001). The euglycemic hyperinsulinemic clamp technique demonstrated that CS treatment improves insulin sensitivity in peripheral tissues. We conclude that the antihyperglycemic effet CS is probably due to the inhibition of basal endogenous glucose production and the improvement of insulin sensitivity in MLDS diabetic mice.

  18. Effects of Encapsulated Propolis on Blood Glycemic Control, Lipid Metabolism, and Insulin Resistance in Type 2 Diabetes Mellitus Rats

    Directory of Open Access Journals (Sweden)

    Yajing Li

    2012-01-01

    Full Text Available The present study investigates the encapsulated propolis on blood glycemic control, lipid metabolism, and insulin resistance in type 2 diabetes mellitus (T2DM rats. The animal characteristics and biological assays of body weight, fasting blood glucose (FBG, fasting serum insulin (FINS, insulin act index (IAI, triglycerides (TG, total cholesterol (TC, high-density lipoprotein cholesterol (HDL-C, and low-density lipoprotein cholesterol (LDL-C were measured and euglycemic hyperinsulinemic glucose clamp technique were used to determine these effects. Our findings show that oral administration of encapsulated propolis can significantly inhibit the increasing of FBG and TG in T2DM rats and can improve IAI and M value in euglycemic hyperinsulinemic clamp experiment. There was no significant effects on body weight, TC, HDL-C, and LDL-C in T2DM rats treated with encapsulated propolis. In conclusion, the results indicate that encapsulated propolis can control blood glucose, modulate lipid metabolism, and improve the insulin sensitivity in T2DM rats.

  19. Substrate Metabolism and Insulin Sensitivity During Fasting in Obese Human Subjects: Impact of GH Blockade.

    Science.gov (United States)

    Pedersen, Morten Høgild; Svart, Mads Vandsted; Lebeck, Janne; Bidlingmaier, Martin; Stødkilde-Jørgensen, Hans; Pedersen, Steen Bønløkke; Møller, Niels; Jessen, Niels; Jørgensen, Jens O L

    2017-04-01

    Insulin resistance and metabolic inflexibility are features of obesity and are amplified by fasting. Growth hormone (GH) secretion increases during fasting and GH causes insulin resistance. To study the metabolic effects of GH blockade during fasting in obese subjects. Nine obese males were studied thrice in a randomized design: (1) after an overnight fast (control), (2) after 72 hour fasting (fasting), and (3) after 72 hour fasting with GH blockade (pegvisomant) [fasting plus GH antagonist (GHA)]. Each study day consisted of a 4-hour basal period followed by a 2-hour hyperinsulinemic, euglycemic clamp combined with indirect calorimetry, assessment of glucose and palmitate turnover, and muscle and fat biopsies. GH levels increased with fasting (P fasting-induced reduction of serum insulin-like growth factor I was enhanced by GHA (P Fasting increased lipolysis and lipid oxidation independent of GHA, but fasting plus GHA caused a more pronounced suppression of lipid intermediates in response to hyperinsulinemic, euglycemic clamp. Fasting-induced insulin resistance was abrogated by GHA (P Fasting plus GHA also caused elevated glycerol levels and reduced levels of counterregulatory hormones. Fasting significantly reduced the expression of antilipolytic signals in adipose tissue independent of GHA. Suppression of GH activity during fasting in obese subjects reverses insulin resistance and amplifies insulin-stimulated suppression of lipid intermediates, indicating that GH is an important regulator of substrate metabolism, insulin sensitivity, and metabolic flexibility also in obese subjects.

  20. Cryogenic Clamp-on Ultrasonic Flowmeters using Single Crystal Piezoelectric Transducers Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Clamp-on ultrasound cryogenic flowmeters using single crystal piezoelectric transducers are proposed to enable reliable, accurate cryogenic instrumentation needs in...

  1. COMPARISON OF EARLY AND DELAYED UMBILICAL CORD CLAMPING IN THE THIRD STAGE OF LABOUR

    Directory of Open Access Journals (Sweden)

    Tamara Serdinšek

    2015-09-01

    Full Text Available Background: Umbilical cord clamping in the third stage of labour is still controversial. Early cord clamping is defined as clamping at 10, 15, 30 or 60 seconds after delivery and delayed as clamping after 60 seconds or at 2-5 minutes after delivery, when the cord stops pulsating or when the placenta is visible within the birth canal. Early clamping is one of the three components of active management of the third stage of labour, which has been used widely in modern obstetrics during the last century. However, in some northern European countries, various parts of the USA and Canada and in developing countries physiological management is preferred.Conclusions: After publication of several trials describing advantages of delayed clamping, this has recently been progressively replacing early clamping. The most important advantages of delayed cord clamping are higher haemoglobin and ferritin levels, higher iron stores, lower incidence of iron deficiency anaemia, better cardiopulmonary adaptation, lower rate of respiratory distress syndrome, and longer duration of early breastfeeding in term neonates, while there is no increase in the incidence of postpartum haemorrhage. Delayed clamping seems to bring some advantages for preterm neonates as well. However, caution is still advised because of the potential adverse effects, especially polycythaemia with hyperviscosity, hyperbilirubinaemia and respiratory distress.

  2. Electrothermally actuated tunable clamped-guided resonant microbeams

    KAUST Repository

    Alcheikh, Nouha

    2017-06-11

    We present simulation and experimental investigation demonstrating active alteration of the resonant and frequency response behavior of resonators by controlling the electrothermal actuation method on their anchors. In-plane clamped-guided arch and straight microbeams resonators are designed and fabricated with V-shaped electrothermal actuators on their anchors. These anchors not only offer various electrothermal actuation options, but also serve as various mechanical stiffness elements that affect the operating resonance frequency of the structures. We have shown that for an arch, the first mode resonance frequency can be increased up to 50% of its initial value. For a straight beam, we have shown that before buckling, the resonance frequency decreases to very low values and after buckling, it increases up to twice of its initial value. These results can be promising for the realization of different wide–range tunable microresonator. The experimental results have been compared to multi-physics finite-element simulations showing good agreement among them.

  3. Electrothermally actuated tunable clamped-guided resonant microbeams

    Science.gov (United States)

    Alcheikh, N.; Hajjaj, A. Z.; Jaber, N.; Younis, M. I.

    2018-01-01

    We present simulation and experimental investigation demonstrating active alteration of the resonant and frequency response behavior of resonators by controlling the electrothermal actuation method on their anchors. In-plane clamped-guided arch and straight microbeams resonators are designed and fabricated with V-shaped electrothermal actuators on their anchors. These anchors not only offer various electrothermal actuation options, but also serve as various mechanical stiffness elements that affect the operating resonance frequency of the structures. We have shown that for an arch, the first mode resonance frequency can be increased up to 50% of its initial value. For a straight beam, we have shown that before buckling, the resonance frequency decreases to very low values and after buckling, it increases up to twice of its initial value. These results can be promising for the realization of different wide-range tunable microresonator. The experimental results have been compared to multi-physics finite-element simulations showing good agreement among them.

  4. Trans-Z-source Neutral Point Clamped inverter

    DEFF Research Database (Denmark)

    Mo, W.; Loh, P. C.; Li, D.

    2012-01-01

    Transformer based Z-source (trans-Z-source) inverters are recently proposed by extending the traditional Z-source inverter with higher buck-boost capability as well as reducing the passive components at the same time. Multi-Level Z-source inverters are single-stage topological solutions used...... for buck-boost energy conversion with all the favourable advantages of multi-level switching retained. This paper presents three-level trans-Z-source Neutral Point Clamped (NPC) inverter topology, which achieves both the advantages of trans-Z-source and three-level NPC inverter configuration. With proper...... modulation scheme, the three-level trans-Z-source inverter can function with minimum of six device commutations per half carrier cycle (same as the traditional buck NPC inverter), while maintaining to produce the designed volt-sec average and inductive voltage boosting at ac output terminals. The designed...

  5. Pulsewidth-modulated 2-source neutral-point-clamped inverter

    DEFF Research Database (Denmark)

    Blaabjerg, Frede; Loh, Poh Chang; Gao, Feng

    2007-01-01

    set of carrier-based modulation expressions can be used for controlling the -source two-level inverter and NPC inverter with and without reduced common-mode switching. All findings presented in this paper have been confirmed in simulation and experimentally using an implemented laboratory prototype.......This paper presents the careful integration of a newly proposed Z-source topological concept to the basic neutral-point-clamped (NPC) inverter topology for designing a three-level inverter with both voltage-buck and voltage-boost capabilities. The designed Z-source NPC inverter uses two unique X......-shaped inductance-capacitance (LC) impedance networks that are connected between two isolated dc input power sources and its inverter circuitry for boosting its AC output voltage. Through the design of an appropriate pulsewidth-modulation (PWM) algorithm, the two impedance networks can be short...

  6. Renal Vascular Clamp Placement: A Potential Cause of Incomplete Hilar Control during Partial Nephrectomy.

    Science.gov (United States)

    Tryon, David; Myklak, Kristene; Alsyouf, Muhannad; Conceicao, Carol; Peplinski, Brandon; Arenas, Javier L; Faaborg, Daniel; Ruckle, Herbert C; Baldwin, D Duane

    2016-03-01

    Previous benchtop studies have shown that robotic bulldog clamps provide incomplete vascular control of a Penrose drain. We determined the efficacy of robotic and laparoscopic bulldog clamps to ensure hemostasis on the human renal artery. The effect of clamp position on vascular control was also examined. Fresh human cadaveric renal arteries were used to determine the leak point pressure of 7 bulldog clamps from a total of 3 manufacturers. Five trials were performed per clamp at 4 locations, including the fulcrum, proximal, middle and distal positions. Comparison was done using the Kruskal-Wallis test with p clamps leaked at a pressure less than 215 mm Hg when applied at the proximal, middle or distal position. In general leak point pressure decreased as the artery was positioned more distal along the clamp. The exception was when the vessel was placed at the fulcrum position. At that position 80% to 100% of trials with the Klein laparoscopic, 100% with the Klein robotic (Klein Robotic, San Antonio, Texas) and 60% to 80% with the Scanlan robotic (Scanlan International, Saint Paul, Minnesota) clamp leaked at pressure below 215 mm Hg. Each vascular clamp adequately occluded flow at physiological pressure when placed at the proximal, middle or distal position. Furthermore, these results demonstrate that there is leakage at physiological pressure when the artery is placed at the fulcrum of certain clamp types. These results suggest that applying a bulldog clamp at the fulcrum could potentially lead to inadequate vessel occlusion and intraoperative bleeding. Copyright © 2016 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

  7. Euglycaemic glucose clamp: what it can and cannot do, and how to do it.

    Science.gov (United States)

    Heise, Tim; Zijlstra, Eric; Nosek, Leszek; Heckermann, Sascha; Plum-Mörschel, Leona; Forst, Thomas

    2016-10-01

    The hyperinsulinaemic-euglycaemic glucose clamp has always been regarded as the "gold standard" for the assessment of pharmacodynamic (PD) properties of insulin preparations; however, there has been controversy over a variety of methodogical details, such as study population, dosing time and the initial stabilization of blood glucose (BG) concentrations at the clamp target level, among clamp groups. As the impact of these details on PD results is unclear, the present review provides an overview of different methodological approaches for both the manual and the automated hyperinsulinaemic-euglycaemic glucose clamp. The advantages and limitations of several methodological details are discussed as well as the relevance of clamp results for the prediction of clinical outcomes. Overall, the best method strongly depends on the exact objective of the trial. If, for instance, duration of action is the primary objective, studies should be carried out in patients with type 1 diabetes to avoid any interference of endogenous insulin. This is less important for variables such as onset of action or early metabolic activity. The hyperinsulinaemic-euglycaemic glucose clamp has a high sensitivity to detect even minor differences between different insulin preparations. The practical relevance of potential differences, however, needs to be investigated in clinical studies. A major prerequisite for obtaining reliable glucose clamp results is the attainment of high clamp quality (i.e. keeping BG concentrations close to the clamp target throughout the experiments). Unfortunately, measures of clamp quality are often under-reported, as is the variability in PD profiles, although these might explain some unconfirmed extreme results obtained in a few clamp studies. © 2016 John Wiley & Sons Ltd.

  8. Application of active electrode compensation to perform continuous voltage-clamp recordings with sharp microelectrodes

    Science.gov (United States)

    Gómez-González, J. F.; Destexhe, A.; Bal, T.

    2014-10-01

    Objective. Electrophysiological recordings of single neurons in brain tissues are very common in neuroscience. Glass microelectrodes filled with an electrolyte are used to impale the cell membrane in order to record the membrane potential or to inject current. Their high resistance induces a high voltage drop when passing current and it is essential to correct the voltage measurements. In particular, for voltage clamping, the traditional alternatives are two-electrode voltage-clamp technique or discontinuous single electrode voltage-clamp (dSEVC). Nevertheless, it is generally difficult to impale two electrodes in a same neuron and the switching frequency is limited to low frequencies in the case of dSEVC. We present a novel fully computer-implemented alternative to perform continuous voltage-clamp recordings with a single sharp-electrode. Approach. To reach such voltage-clamp recordings, we combine an active electrode compensation algorithm (AEC) with a digital controller (AECVC). Main results. We applied two types of control-systems: a linear controller (proportional plus integrative controller) and a model-based controller (optimal control). We compared the performance of the two methods to dSEVC using a dynamic model cell and experiments in brain slices. Significance. The AECVC method provides an entirely digital method to perform continuous recording and smooth switching between voltage-clamp, current clamp or dynamic-clamp configurations without introducing artifacts.

  9. A structural and mechanistic study of π-clamp-mediated cysteine perfluoroarylation.

    Science.gov (United States)

    Dai, Peng; Williams, Jonathan K; Zhang, Chi; Welborn, Matthew; Shepherd, James J; Zhu, Tianyu; Van Voorhis, Troy; Hong, Mei; Pentelute, Bradley L

    2017-08-11

    Natural enzymes use local environments to tune the reactivity of amino acid side chains. In searching for small peptides with similar properties, we discovered a four-residue π-clamp motif (Phe-Cys-Pro-Phe) for regio- and chemoselective arylation of cysteine in ribosomally produced proteins. Here we report mutational, computational, and structural findings directed toward elucidating the molecular factors that drive π-clamp-mediated arylation. We show the significance of a trans conformation prolyl amide bond for the π-clamp reactivity. The π-clamp cysteine arylation reaction enthalpy of activation (ΔH‡) is significantly lower than a non-π-clamp cysteine. Solid-state NMR chemical shifts indicate the prolyl amide bond in the π-clamp motif adopts a 1:1 ratio of the cis and trans conformation, while in the reaction product Pro3 was exclusively in trans. In two structural models of the perfluoroarylated product, distinct interactions at 4.7 Å between Phe1 side chain and perfluoroaryl electrophile moiety are observed. Further, solution 19F NMR and isothermal titration calorimetry measurements suggest interactions between hydrophobic side chains in a π-clamp mutant and the perfluoroaryl probe. These studies led us to design a π-clamp mutant with an 85-fold rate enhancement. These findings will guide us toward the discovery of small reactive peptides to facilitate abiotic chemistry in water.

  10. Application of active electrode compensation to perform continuous voltage-clamp recordings with sharp microelectrodes.

    Science.gov (United States)

    Gómez-González, J F; Destexhe, A; Bal, T

    2014-10-01

    Electrophysiological recordings of single neurons in brain tissues are very common in neuroscience. Glass microelectrodes filled with an electrolyte are used to impale the cell membrane in order to record the membrane potential or to inject current. Their high resistance induces a high voltage drop when passing current and it is essential to correct the voltage measurements. In particular, for voltage clamping, the traditional alternatives are two-electrode voltage-clamp technique or discontinuous single electrode voltage-clamp (dSEVC). Nevertheless, it is generally difficult to impale two electrodes in a same neuron and the switching frequency is limited to low frequencies in the case of dSEVC. We present a novel fully computer-implemented alternative to perform continuous voltage-clamp recordings with a single sharp-electrode. To reach such voltage-clamp recordings, we combine an active electrode compensation algorithm (AEC) with a digital controller (AECVC). We applied two types of control-systems: a linear controller (proportional plus integrative controller) and a model-based controller (optimal control). We compared the performance of the two methods to dSEVC using a dynamic model cell and experiments in brain slices. The AECVC method provides an entirely digital method to perform continuous recording and smooth switching between voltage-clamp, current clamp or dynamic-clamp configurations without introducing artifacts.

  11. Implementing dynamic clamp with synaptic and artificial conductances in mouse retinal ganglion cells.

    Science.gov (United States)

    Huang, Jin Y; Stiefel, Klaus M; Protti, Dario A

    2013-05-16

    Ganglion cells are the output neurons of the retina and their activity reflects the integration of multiple synaptic inputs arising from specific neural circuits. Patch clamp techniques, in voltage clamp and current clamp configurations, are commonly used to study the physiological properties of neurons and to characterize their synaptic inputs. Although the application of these techniques is highly informative, they pose various limitations. For example, it is difficult to quantify how the precise interactions of excitatory and inhibitory inputs determine response output. To address this issue, we used a modified current clamp technique, dynamic clamp, also called conductance clamp (1, 2, 3) and examined the impact of excitatory and inhibitory synaptic inputs on neuronal excitability. This technique requires the injection of current into the cell and is dependent on the real-time feedback of its membrane potential at that time. The injected current is calculated from predetermined excitatory and inhibitory synaptic conductances, their reversal potentials and the cell's instantaneous membrane potential. Details on the experimental procedures, patch clamping cells to achieve a whole-cell configuration and employment of the dynamic clamp technique are illustrated in this video article. Here, we show the responses of mouse retinal ganglion cells to various conductance waveforms obtained from physiological experiments in control conditions or in the presence of drugs. Furthermore, we show the use of artificial excitatory and inhibitory conductances generated using alpha functions to investigate the responses of the cells.

  12. The transducer domain is important for clamp operation in human DNA topoisomerase IIalpha

    DEFF Research Database (Denmark)

    Oestergaard, Vibe H; Bjergbaek, Lotte; Skouboe, Camilla

    2003-01-01

    the clamp to the enzyme core. Although structurally conserved, it is unclear whether the transducer domain is involved in clamp mechanism. We have purified and characterized a human topoisomerase II alpha enzyme with a two-amino acid insertion at position 408 in the transducer domain. The enzyme retains...

  13. Using atomic force microscopy to investigate patch-clamped nuclear membrane.

    Science.gov (United States)

    Danker, T; Mazzanti, M; Tonini, R; Rakowska, A; Oberleithner, H

    1997-11-01

    Nuclear patch clamp is an emerging research field that aims to disclose the electrical phenomena underlying macromolecular transport across the nuclear envelope (NE), its properties as an ion barrier and its function as an intracellular calcium store. The authors combined the patch clamp technique with atomic force microscopy (AFM) to investigate the structure-function relationship of NE. In principle, patch clamp currents, recorded from the NE can indicate the activity of the nuclear pore complexes (NPCs) and/or of ion channels in the two biomembranes that compose the NE. However, the role of the NPCs is still nuclear because the observed NE current in patch clamp experiments is lower than expected from the known density of the NPCs. Therefore, AFM was applied to link patch clamp currents to structure. The membrane patch was excised from the nuclear envelope and, after electrical evaluation, transferred from the patch pipette to a substrate. We could identify the native nuclear membrane patches with AFM at a lateral and a vertical resolution of 3 nm and 0.1 nm, respectively. It was shown that complete NE together with NPCs can be excised from the nucleus after their functional identification in patch clamp experiments. However, we also show that membranes of the endoplasmic reticulum can contaminate the tip of the patch pipette during nuclear patch clamp experiments. This possibility must be considered carefully in nuclear patch clamp experiments.

  14. Relating glucose clamp profiles to reduction of blood glucose after insulin administration.

    Science.gov (United States)

    Woodworth, James R; Howey, Daniel C; Bowsher, Ronald R; Brunelle, Rocco L

    2004-04-01

    A model was established allowing prediction of blood glucose response from glucose clamp results performed in healthy volunteers. Data from published studies performed in healthy volunteers were used to establish, test, and validate a model for the evaluation of glucose reductions from glucose clamp results. Studies included those that measured blood glucose and glucodynamic response over time after administration of 0.05 U/kg of regular human insulin (HR) and insulin lispro (LP) with and without the benefit of a glucose clamp procedure. An inhibitory effect E(max) model was used to describe the relationship; the model differed between the HR and LP responses by the intensity of the counterregulatory response as assessed by glucagon measurements. The relationships were used to predict blood glucose responses from a clamp study assessing NPH insulin and HR administrations. Glucose concentrations measured after administration of NPH insulin and HR without a clamp were compared to the model-predicted results to assess the accuracy of the model predictions. The E(max) model successfully correlated the glucose clamp results with the blood glucose depressions in the presence and absence of a counterregulatory response. However, predictions of glucose depression were only accurately modeled in the absence of a counterregulatory glucagon response. The correlations established with a minimal counterregulatory response underscore the value of glucose clamp procedures in defining the time-activity profiles of insulins when the clamp is established at fasting glucose concentrations.

  15. Modeling of SVM Diode Clamping Three-Level Inverter Connected to Grid

    DEFF Research Database (Denmark)

    Guo, Yougui; Zeng, Ping; Zhu, Jieqiong

    2011-01-01

    PLECS is used to model the diode clamping three-level inverter connected to grid and good results are obtained. First the output voltage SVM is described for diode clamping three-level inverter with loads connected to Y. Then the output voltage SVM of diode clamping three-level inverter is simply...... analyzed with loads connected to △. But it will be further researched in the future. Third, PLECS is briefly introduced. Fourth, the modeling of diode clamping three-level inverter is briefly presented with PLECS. Finally, a series of simulations are carried out. The simulation results tell us PLECS...... is very powerful tool to real power circuits and it is very easy to simulate them. They have also verified that SVM control strategy is feasible to control the diode clamping three-level inverter....

  16. A reduction clamp for an aiming component in associated acetabular fractures

    Directory of Open Access Journals (Sweden)

    Zhang-Fu Wang

    2015-01-01

    Full Text Available Background: The treatment of acetabular fractures is complex and requires specialized equipment. However, all currently available instruments have some disadvantages. A new reduction clamp that can firmly enable reduction and not hinder subsequent fixation procedures for some special fracture types is needed. Materials and Methods: In this study, we introduce a new acetabular clamp and its preliminary clinical application in three T-shaped acetabular fractures. Results: This new clamp can successfully pull the posterior column back to the anterior column and firmly maintain the reduction. This clamp′s aiming plate can facilitate the insertion of long lag screws. The clamp is also easy to assemble and use. Conclusion: This reduction clamp is a useful instrument that can facilitate open reduction and internal fixation of acetabular fractures.

  17. Escherichia coli processivity clamp β from DNA polymerase III is dynamic in solution†

    Science.gov (United States)

    Fang, Jing; Engen, John R.; Beuning, Penny J.

    2011-01-01

    Escherichia coli DNA polymerase III is a highly processive replicase due to the presence of the β clamp protein that tethers DNA polymerases to DNA. The β clamp is a head-to-tail ring-shaped homodimer, in which each protomer contains three structurally similar domains. Although multiple studies have probed the functions of the β clamp, a detailed understanding of the conformational dynamics of the β clamp in solution is lacking. Here we used hydrogen exchange mass spectrometry to characterize the conformation and dynamics of the intact dimer β clamp and a variant form (I272A/L273A) with diminished ability to dimerize in solution. Our data indicate that the β clamp is not a static closed ring but rather is dynamic in solution. The three domains showed different dynamics though they share a highly similar tertiary structure. Domain I, which controls the opening of the clamp by dissociating from Domain III, contained several highly flexible peptides that underwent partial cooperative unfolding (EX1 kinetics) with a half-life ~4 h. The comparison between the β monomer variant and the wild-type β clamp showed that the β monomer was more dynamic. In the monomer, partial unfolding was much faster and additional regions of Domain III also underwent partial unfolding with a half-life ~1 h. Our results suggest that the δ subunit of the clamp loader may function as a “ring holder” to stabilize the transient opening of the β clamp, rather than as a “ring opener”. PMID:21657794

  18. Generalization of the dynamic clamp concept in neurophysiology and behavior.

    Directory of Open Access Journals (Sweden)

    Pablo Chamorro

    Full Text Available The idea of closed-loop interaction in in vitro and in vivo electrophysiology has been successfully implemented in the dynamic clamp concept strongly impacting the research of membrane and synaptic properties of neurons. In this paper we show that this concept can be easily generalized to build other kinds of closed-loop protocols beyond (or in addition to electrical stimulation and recording in neurophysiology and behavioral studies for neuroethology. In particular, we illustrate three different examples of goal-driven real-time closed-loop interactions with drug microinjectors, mechanical devices and video event driven stimulation. Modern activity-dependent stimulation protocols can be used to reveal dynamics (otherwise hidden under traditional stimulation techniques, achieve control of natural and pathological states, induce learning, bridge between disparate levels of analysis and for a further automation of experiments. We argue that closed-loop interaction calls for novel real time analysis, prediction and control tools and a new perspective for designing stimulus-response experiments, which can have a large impact in neuroscience research.

  19. Non-Hookean statistical mechanics of clamped graphene ribbons

    Science.gov (United States)

    Bowick, Mark J.; Košmrlj, Andrej; Nelson, David R.; Sknepnek, Rastko

    2017-03-01

    Thermally fluctuating sheets and ribbons provide an intriguing forum in which to investigate strong violations of Hooke's Law: Large distance elastic parameters are in fact not constant but instead depend on the macroscopic dimensions. Inspired by recent experiments on free-standing graphene cantilevers, we combine the statistical mechanics of thin elastic plates and large-scale numerical simulations to investigate the thermal renormalization of the bending rigidity of graphene ribbons clamped at one end. For ribbons of dimensions W ×L (with L ≥W ), the macroscopic bending rigidity κR determined from cantilever deformations is independent of the width when W expected. When W >ℓth , however, this thermally renormalized bending rigidity begins to systematically increase, in agreement with the scaling theory, although in our simulations we were not quite able to reach the system sizes necessary to determine the fully developed power law dependence on W . When the ribbon length L >ℓp , where ℓp is the W -dependent thermally renormalized ribbon persistence length, we observe a scaling collapse and the beginnings of large scale random walk behavior.

  20. Indentation metrology of clamped, ultra-thin elastic sheets.

    Science.gov (United States)

    Vella, Dominic; Davidovitch, Benny

    2017-03-15

    We study the indentation of ultrathin elastic sheets clamped to the edge of a circular hole. This classical setup has received considerable attention lately, being used by various experimental groups as a probe to measure the surface properties and stretching modulus of thin solid films. Despite the apparent simplicity of this method, the geometric nonlinearity inherent in the mechanical response of thin solid objects renders the analysis of the resulting data a nontrivial task. Importantly, the essence of this difficulty is in the geometric coupling between in-plane stress and out-of-plane deformations, and hence is present in the behaviour of Hookean solids even when the slope of the deformed membrane remains small. Here we take a systematic approach to address this problem, using the membrane limit of the Föppl-von-Kármán equations. This approach highlights some of the dangers in the use of approximate formulae in the metrology of solid films, which can introduce large errors; we suggest how such errors may be avoided in performing experiments and analyzing the resulting data.

  1. Deletion of GPR40 Impairs Glucose-Induced Insulin Secretion In Vivo in Mice Without Affecting Intracellular Fuel Metabolism in Islets

    Energy Technology Data Exchange (ETDEWEB)

    Alquier, Thierry; Peyot, Marie-Line; Latour, M. G.; Kebede, Melkam; Sorensen, Christina M.; Gesta, Stephane; Kahn, C. R.; Smith, Richard D.; Jetton, Thomas L.; Metz, Thomas O.; Prentki, Marc; Poitout, Vincent J.

    2009-11-01

    The G protein-coupled receptor GPR40 mediates fatty-acid potentiation of glucose-stimulated insulin secretion, but its contribution to insulin secretion in vivo and mechanisms of action remain uncertain. This study was aimed to ascertain whether GPR40 controls insulin secretion in vivo and modulates intracellular fuel metabolism in islets. We observed that glucose- and arginine-stimulated insulin secretion, assessed by hyperglycemic clamps, was decreased by approximately 60% in GPR40 knock-out (KO) fasted and fed mice, without changes in insulin sensitivity assessed by hyperinsulinemic-euglycemic clamps. Glucose and palmitate metabolism were not affected by GPR40 deletion. Lipid profiling revealed a similar increase in triglyceride and decrease in lysophosphatidylethanolamine species in WT and KO islets in response to palmitate. These results demonstrate that GPR40 regulates insulin secretion in vivo not only in response to fatty acids but also to glucose and arginine, without altering intracellular fuel metabolism.

  2. Effect of arotinolol on insulin sensitivity in obese hypertensive patients.

    Science.gov (United States)

    Kageyama, S; Yamamoto, J; Mimura, A; Sakurai, T; Ishibashi, K; Isogai, Y

    1994-01-01

    Insulin sensitivity in terms of glucose disposal rate was determined by using the hyperinsulinemic euglycemic clamp technique in seven obese hypertensive patients and nine normotensive control subjects. The technique was used on all study subjects at baseline and on the hypertensive patients after 12 weeks of arotinolol treatment. Mean supine systolic blood pressure decreased from 155 +/- 3 mm Hg before treatment to 133 +/- 6 mm Hg after treatment (P arotinolol was 10 mg/d, except for one patient who received 20 mg/d. The glucose disposal rate during the last 30 minutes of the glucose clamp procedure was not significantly affected by treatment with arotinolol. Compared with the hypertensive patients, the glucose disposal rate in the normotensive subjects was significantly increased, both before and after treatment (P arotinolol did not significantly affect insulin sensitivity.

  3. Insulin sensitivity and metabolic flexibility following exercise training among different obese insulin resistant phenotypes

    DEFF Research Database (Denmark)

    Malin, Steven K; Haus, Jacob M; Solomon, Thomas

    2013-01-01

    Impaired fasting glucose (IFG) blunts the reversal of impaired glucose tolerance (IGT) after exercise training. Metabolic inflexibility has been implicated in the etiology of insulin resistance, however, the efficacy of exercise on peripheral and hepatic insulin sensitivity or substrate utilization......) and were instructed to maintain a eucaloric diet. A euglycemic-hyperinsulinemic clamp (40 mU/m(2)/min) with [6,6-(2)H]-glucose was used to determine peripheral and hepatic insulin sensitivity. Non-oxidative glucose disposal and metabolic flexibility (insulin-stimulated respiratory quotient [RQ] minus...... fasting RQ) were also assessed. Glucose incremental area under the curve was calculated from the OGTT (iAUCOGTT). Exercise increased clamp-derived peripheral and hepatic insulin sensitivity more in adults with IFG or IGT alone than IFG+IGT (P...

  4. Effect of gender on lipid-induced insulin resistance in obese subjects

    DEFF Research Database (Denmark)

    Vistisen, Bodil; Hellgren, Lars; Vadset, T.

    2008-01-01

    Objective: In obese subjects, chronically elevated plasma concentrations of non-esterified fatty acids (NEFAs) exert a marked risk to contract insulin resistance and subsequently type 2 diabetes. When NEFA is acutely increased due to i.v. infusion of lipid, glucose disposal during...... a hyperinsulinemic-euglycemic clamp is reduced. This effect has been explained by a NEFA-induced decrease in skeletal muscle insulin sensitivity caused by accumulation of the lipid intermediates Such as ceramide and diacylglycerol in the myocytes. However, neither the lipid-induced reduction of glucose disposal nor...... the clamp was similar in females and males (46+/-10 and 60+/-4%,, respectively, NS). However, whole-body insulin sensitivity as well as non-oxidative glucose disposal was higher in obese females compared with obese males both during lipid and saline infusion (P...

  5. Sortilin 1 knockout alters basal adipose glucose metabolism but not diet-induced obesity in mice.

    Science.gov (United States)

    Li, Jibiao; Matye, David J; Wang, Yifeng; Li, Tiangang

    2017-04-01

    Sortilin 1 (Sort1) is a trafficking receptor that has been implicated in the regulation of plasma cholesterol in humans and mice. Here, we use metabolomics and hyperinsulinemic-euglycemic clamp approaches to obtain further understanding of the in vivo effects of Sort1 deletion on diet-induced obesity as well as on adipose lipid and glucose metabolism. Results show that Sort1 knockout (KO) does not affect Western diet-induced obesity nor adipose fatty acid and ceramide concentrations. Under the basal fasting state, chow-fed Sort1 KO mice have decreased adipose glycolytic metabolites, but Sort1 deletion does not affect insulin-stimulated tissue glucose uptake during the insulin clamp. These results suggest that Sort1 loss-of-function in vivo does not affect obesity development, but differentially modulates adipose glucose metabolism under fasting and insulin-stimulated states. © 2017 Federation of European Biochemical Societies.

  6. Supraceliac, but not infrarenal, aortic cross-clamping upregulates neutrophil integrin CD11b.

    Science.gov (United States)

    Hill, G E; Mihalakakos, P J; Spurzem, J R; Baxter, T B

    1995-10-01

    To evaluate the effects of supraceliac and infrarenal aortic cross-clamping on the expression of neutrophil integrin in CD11b (a marker of systemic cytokine release). Two groups, determined by anatomic placement of aortic cross-clamp. Laboratory personnel were blinded as to group assignment. University teaching and community hospitals. Laboratory facilities used were university and Veteran's Affairs medical centers. Patients scheduled for aortic surgery. Blood sampling was performed at baseline, after 30 minutes of aortic cross-clamp duration, 30 and 90 minutes after reperfusion (for tumor necrosis factor-alpha plasma levels in infrarenal cross-clamp group), and at baseline and 90 minutes reperfusion (for neutrophil CD11b expression quantification) in both groups. Tumor necrosis factor-alpha measured by ELISA technique did not change at any time period in the infrarenal clamping group. Neutrophil CD11b expression, measured by double antibody staining and FACScan analysis, did not change significantly at 90 minutes of reperfusion in the infrarenal group, but increased significantly (p entrapment and subsequent neutrophil-mediated reperfusion injury. These results suggest that upregulation of neutrophil integrin CD11b after supraceliac aortic clamping may in part be responsible for the higher incidence of acute lung injury after thoracic aortic aneurysm repair requiring supraceliac clamping when compared with infrarenal aneurysm surgery.

  7. Laparoscopic Partial Nephrectomy with a Remotely Activated Bulldog Clamp in a Pig Model.

    Science.gov (United States)

    Chen, Wei; Guo, Jian-Ming; Hou, Jun; Wang, Guo-Min

    2017-01-01

    Saving warm ischemia time (WIT) in laparoscopic partial nephrectomy (LPN) is important. We have designed a new device, a remotely activated bulldog clamp, to facilitate LPN. In present study, the effectiveness, convenience, and safety of the new design device were compared with that of standard bulldog vascular clamp in a pig model. Fifteen cases of LPN were bilaterally operated on upper or lower pole of kidneys in five pigs. As the test group, eight cases involved the remotely activated bulldog clamp, whereas the remaining seven cases, as the control group, involved the common standard vascular bulldog clamp. The mean operative time, blood lost, and mean WIT were compared between the two groups. The mean operative time and blood lost were not significantly different between the groups (p clamping was used in the two groups. With on-demand clamping, the mean WIT in the test group was 19.0 ± 3.42 minutes, which was significantly less compared with the control group (31.54 ± 5.23 minutes, p clamping and unclamping much easier. It is effective and safe in LPN.

  8. Dynamic assembly of Hda and the sliding clamp in the regulation of replication licensing.

    Science.gov (United States)

    Kim, Jin S; Nanfara, Michael T; Chodavarapu, Sundari; Jin, Kyeong S; Babu, Vignesh M P; Ghazy, Mohamed A; Chung, Scisung; Kaguni, Jon M; Sutton, Mark D; Cho, Yunje

    2017-04-20

    Regulatory inactivation of DnaA (RIDA) is one of the major regulatory mechanisms of prokaryotic replication licensing. In RIDA, the Hda-sliding clamp complex loaded onto DNA directly interacts with adenosine triphosphate (ATP)-bound DnaA and stimulates the hydrolysis of ATP to inactivate DnaA. A prediction is that the activity of Hda is tightly controlled to ensure that replication initiation occurs only once per cell cycle. Here, we determined the crystal structure of the Hda-β clamp complex. This complex contains two pairs of Hda dimers sandwiched between two β clamp rings to form an octamer that is stabilized by three discrete interfaces. Two separate surfaces of Hda make contact with the β clamp, which is essential for Hda function in RIDA. The third interface between Hda monomers occludes the active site arginine finger, blocking its access to DnaA. Taken together, our structural and mutational analyses of the Hda-β clamp complex indicate that the interaction of the β clamp with Hda controls the ability of Hda to interact with DnaA. In the octameric Hda-β clamp complex, the inability of Hda to interact with DnaA is a novel mechanism that may regulate Hda function. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  9. Planar patch clamp for neuronal networks--considerations and future perspectives.

    Science.gov (United States)

    Bosca, Alessandro; Martina, Marzia; Py, Christophe

    2014-01-01

    The patch-clamp technique is generally accepted as the gold standard for studying ion channel activity allowing investigators to either "clamp" membrane voltage and directly measure transmembrane currents through ion channels, or to passively monitor spontaneously occurring intracellular voltage oscillations. However, this resulting high information content comes at a price. The technique is labor-intensive and requires highly trained personnel and expensive equipment. This seriously limits its application as an interrogation tool for drug development. Patch-clamp chips have been developed in the last decade to overcome the tedious manipulations associated with the use of glass pipettes in conventional patch-clamp experiments. In this chapter, we describe some of the main materials and fabrication protocols that have been developed to date for the production of patch-clamp chips. We also present the concept of a patch-clamp chip array providing high resolution patch-clamp recordings from individual cells at multiple sites in a network of communicating neurons. On this chip, the neurons are aligned with the aperture-probes using chemical patterning. In the discussion we review the potential use of this technology for pharmaceutical assays, neuronal physiology and synaptic plasticity studies.

  10. Blood gas values in clamped and unclamped umbilical cord at birth.

    Science.gov (United States)

    Di Tommaso, Mariarosaria; Seravalli, Viola; Martini, Irene; La Torre, Pasquale; Dani, Carlo

    2014-09-01

    To determine the reliability of the cord blood gas analysis on the unclamped cord compared to the standard technique of sampling on double clamped cord. Prospective observational study conducted on 46 singleton neonates vaginally delivered at term. Matched pairs of umbilical artery and vein blood samples were collected from unclamped cord within 90s after birth and from the same cord after clamping, with the clamping occurring immediately after the first blood collection. A blood gas analysis was performed on each collected sample. Arterial and venous blood samples were analyzed for pH, PO2, pCO2, SaO2, hemoglobin concentration (ctHb) and base excess (BE). The values were compared between the two groups (clamped vs unclamped) using a Wilcoxon test. No significant difference was found in pH, PO2, pCO2, SaO2 and ctHb values on arterial blood between unclamped and clamped cord. The only significant difference was related to BE (pblood, the values of pH, PO2, pCO2 were comparable between unclamped and clamped cord, while the values of SaO2, ctHb and BE were significantly different (pblood gas parameters and in the main venous blood gas parameters between unclamped and clamped cord. Sampling of cord blood for gas analysis may be performed on the unclamped cord right after birth without reducing the accuracy of the analysis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Prognóstico da hipoglicemia hiperinsulinêmica persistente da infância: uma revisão sistemática Prognosis of persistent hyperinsulinemic hypoglycemia of infancy: a systematic review

    Directory of Open Access Journals (Sweden)

    Cláudio Flauzino Oliveira

    2007-09-01

    Full Text Available OBJETIVO: Descrever o prognóstico de crianças com hipoglicemia hiperinsulinêmica persistente da infância, submetidos ou não à pancreatectomia. FONTES DE DADOS: Foram selecionados, por meio das bases de dados Medline e Ovid, estudos publicados nos últimos dez anos contendo o termo "hyperinsulinemic hypoglycemia". A partir desta seleção, foram analisados os artigos que realizaram estudos observacionais sobre o prognóstico da hipoglicemia hiperinsulinêmica persistente da infância (zero a dez anos, para a revisão sistemática. SÍNTESE DOS DADOS: Foram identificadas 269 publicações, das quais 13 continham informações sobre prognóstico das crianças com hipoglicemia hiperinsulinêmica persistente da infância, incluindo oito artigos referentes a pacientes submetidos à pancreatectomia. O principal determinante do prognóstico é o quadro neurológico. Nos estudos apresentados, a incidência de retardo do desenvolvimento neurológico variou de 10 a 70%, dependendo da casuística e da idade de aparecimento dos sintomas. Entre as crianças submetidas à pancreatectomia para controle da hipoglicemia, o principal fator que influenciou o prognóstico em longo prazo foi a característica e a extensão da lesão pancreática. As lesões focais acarretam significativamente menos complicações que as difusas. A incidência de diabetes melito após a pancreatectomia parcial ou subtotal variou de 25 a 100% dos pacientes, de acordo com a casuística avaliada. CONCLUSÕES: O momento de aparecimento dos sintomas influi diretamente sobre a gravidade do quadro neurológico e, consequentemente, sobre o prognóstico. Nos pacientes submetidos à pancreatectomia, a característica da lesão determina maiores ou menores conseqüências.OBJECTIVE: To describe the prognosis of persistent hyperinsulinemic hypoglycemia of infancy, submitted or not to pancreatectomy. DATA SOURCES: Databases Medline and Ovid were searched for studies published in the last ten

  12. Metformin augments the levels of molecules that regulate the expression of the insulin-dependent glucose transporter GLUT4 in the endometria of hyperinsulinemic PCOS patients.

    Science.gov (United States)

    Carvajal, Rodrigo; Rosas, Carlos; Kohan, Karla; Gabler, Fernando; Vantman, David; Romero, Carmen; Vega, Margarita

    2013-08-01

    Does treatment with the insulin sensitizer metformin modify the levels and activation of proteins related to the expression of the insulin-dependent glucose transporter (GLUT4), such as adenosine monophosphate-activated protein kinase (AMPK) and myocyte enhancer factor 2A (MEF2A), in endometria from hyperinsulinemic hyperandrogenemic polycystic ovary syndrome (PCOS h-Ins) patients? In PCOS h-Ins patients, metformin increases endometrial levels of GLUT4 mRNA and protein levels by normalizing the quantity and activation of molecules that regulate GLUT4 expression to healthy values. These changes could improve endometrial metabolic function. PCOS is an endocrine-metabolic disorders closely associated with insulin resistance. In particular, the insulin signaling pathway is impaired in endometria from these patients and the concentration of GLUT4, as well as the molecules involved in its translocation to the cell surface, is decreased. However, there are limited data about the mechanisms that regulate the GLUT4 expression in the endometria and the effect of metformin on them. This is a case-control study in the setting of a research unit, approved by the Ethical Committees of our institution. The groups whose endometria were studied were PCOS h-Ins (n = 8); PCOS patients with hyperandrogenemia hyperinsulinemia taking only metformin for at least 3 months (PCOS-MTF, n = 8) and healthy fertile women at the time of hysterectomy because of benign pathology as controls (CE, n = 8). Steroids and sex hormone-binding globulin were measured and glucose and insulin levels were evaluated during an oral glucose tolerance test. Protein levels for αAMPK (catalytic subunit of AMPK), phosphorylated (p)-AMPKαThr(172) (activating phosphorylation site), MEF2A, p-MEF2AThr312 (activating phosphorylation site) and GLUT4 were assessed by western blot and immunohistochemistry. In addition, GLUT4 gene expression was evaluated by RT-PCR. We found significantly lower levels of MEF2A and p-MEF2

  13. Analysis of the role of PCNA-DNA contacts during clamp loading

    Directory of Open Access Journals (Sweden)

    Goedken Eric R

    2010-01-01

    Full Text Available Abstract Background Sliding clamps, such as Proliferating Cell Nuclear Antigen (PCNA in eukaryotes, are ring-shaped protein complexes that encircle DNA and enable highly processive DNA replication by serving as docking sites for DNA polymerases. In an ATP-dependent reaction, clamp loader complexes, such as the Replication Factor-C (RFC complex in eukaryotes, open the clamp and load it around primer-template DNA. Results We built a model of RFC bound to PCNA and DNA based on existing crystal structures of clamp loaders. This model suggests that DNA would enter the clamp at an angle during clamp loading, thereby interacting with positively charged residues in the center of PCNA. We show that simultaneous mutation of Lys 20, Lys 77, Arg 80, and Arg 149, which interact with DNA in the RFC-PCNA-DNA model, compromises the ability of yeast PCNA to stimulate the DNA-dependent ATPase activity of RFC when the DNA is long enough to extend through the clamp. Fluorescence anisotropy binding experiments show that the inability of the mutant clamp proteins to stimulate RFC ATPase activity is likely caused by reduction in the affinity of the RFC-PCNA complex for DNA. We obtained several crystal forms of yeast PCNA-DNA complexes, measuring X-ray diffraction data to 3.0 Å resolution for one such complex. The resulting electron density maps show that DNA is bound in a tilted orientation relative to PCNA, but makes different contacts than those implicated in clamp loading. Because of apparent partial disorder in the DNA, we restricted refinement of the DNA to a rigid body model. This result contrasts with previous analysis of a bacterial clamp bound to DNA, where the DNA was well resolved. Conclusion Mutational analysis of PCNA suggests that positively charged residues in the center of the clamp create a binding surface that makes contact with DNA. Disruption of this positive surface, which had not previously been implicated in clamp loading function, reduces RFC

  14. Automated Patch Clamp Meets High-Throughput Screening: 384 Cells Recorded in Parallel on a Planar Patch Clamp Module.

    Science.gov (United States)

    Obergrussberger, Alison; Brüggemann, Andrea; Goetze, Tom A; Rapedius, Markus; Haarmann, Claudia; Rinke, Ilka; Becker, Nadine; Oka, Takayuki; Ohtsuki, Atsushi; Stengel, Timo; Vogel, Marius; Steindl, Juergen; Mueller, Max; Stiehler, Johannes; George, Michael; Fertig, Niels

    2016-12-01

    We have developed an automated patch clamp module for high-throughput ion channel screening, recording from 384 cells simultaneously. The module is incorporated into a laboratory pipetting robot and uses a 384-channel pipettor head for application of cells and compounds. The module contains 384 amplifier channels for fully parallel recordings using a digital amplifier. Success rates for completed experiments (1- to 4-point concentration-response curves for cells satisfying defined quality control parameters) of greater than 85% have been routinely achieved with, for example, HEK, CHO, and RBL cell lines expressing hNaV1.7, hERG, Kir2.1, GABA, or glutamate receptors. Pharmacology experiments are recorded and analyzed using specialized software, and the pharmacology of hNaV1.7 and hERG is described. Fast external solution exchange rates of robot to minimize exposure of the ligand on the receptor. This ensures that ligand-gated ion channels, for example, GABA and glutamate described in this report, can be reproducibly recorded. Stem cell-derived cardiomyocytes have also been used with success rates of 52% for cells that have a seal resistance of >200 MΩ, and recordings of voltage-gated Na+ and Ca2+ are shown. © 2015 Society for Laboratory Automation and Screening.

  15. Effects of current clamp on chick retinal pigment epithelium.

    Science.gov (United States)

    Fujii, S; Hughes, B A; Steinberg, R H

    1991-06-01

    The basal membrane of the retinal pigment epithelium (RPE) is the origin of two components of the electroretinogram, the fast oscillation and the light peak. Both of these responses originate from changes in basal membrane potential (Vba), and both are associated with changes in basal membrane resistance (Rba). In addition, many experimental manipulations that alter Vba also produce apparent changes in Rba. These findings raise the possibility that the basal membrane contains a voltage-sensitive conductance that operates in the physiologic range and is involved causally in light-evoked and other responses. We report the results of current clamp experiments on the isolated retina-RPE-choroid of chick that were designed to test for the presence of such a voltage-sensitive conductance in the basal membrane. Depolarizing Vba by 15 mV with retina-to-choroid current had essentially no effect on either the ratio of membrane resistances (Rap/Rba) or the transtissue resistance (RTotal), indicating no alteration in Rba. In contrast, hyperpolarizing Vba by 15 mV with choroid-to-retina current caused a gradual decrease in RTotal and increase in Rap/Rba. Analysis of accompanying changes in membrane voltages and changes in intracellular c-wave amplitude suggested that the most likely cause of the decrease in RTotal is a decrease in paracellular resistance. Voltage-sensitive conductances of the basal membrane appear to play little or no role in the resistance changes that accompany changes in Vba in the physiologic range. The conductance changes underlying the fast oscillation and light peak probably result from either the modulation of channels by second messengers or changes in intracellular ion concentration.

  16. Implementation of a fast 16-Bit dynamic clamp using LabVIEW-RT.

    Science.gov (United States)

    Kullmann, Paul H M; Wheeler, Diek W; Beacom, Joshua; Horn, John P

    2004-01-01

    The dynamic-clamp method provides a powerful electrophysiological tool for creating virtual ionic conductances in living cells and studying their influence on membrane potential. Here we describe G-clamp, a new way to implement a dynamic clamp using the real-time version of the Lab-VIEW programming environment together with a Windows host, an embedded microprocessor that runs a real-time operating system and a multifunction data-acquisition board. The software includes descriptions of a fast voltage-dependent sodium conductance, delayed rectifier, M-type and A-type potassium conductances, and a leak conductance. The system can also read synaptic conductance waveforms from preassembled data files. These virtual conductances can be reliably implemented at speeds LabVIEW should make it relatively easy for others to adapt G-clamp for new experimental applications.

  17. Model Clamp: A Computer Tool to Probe Action Potential Transfer Between Cardiac Cells

    National Research Council Canada - National Science Library

    Wilders, Ronald

    2001-01-01

    In the early nineties, Joyner and coworkers introduced the "coupling clamp" technique in which an isolated cardiac cell can be electrically coupled to either another isolated cardiac cell or to an analog model cell (RC circuit...

  18. Stepwise unfolding of titin under force-clamp atomic force microscopy.

    Science.gov (United States)

    Oberhauser, A F; Hansma, P K; Carrion-Vazquez, M; Fernandez, J M

    2001-01-16

    Here we demonstrate the implementation of a single-molecule force clamp adapted for use with an atomic force microscope. We show that under force-clamp conditions, an engineered titin protein elongates in steps because of the unfolding of its modules and that the waiting times to unfold are exponentially distributed. Force-clamp measurements directly measure the force dependence of the unfolding probability and readily captures the different mechanical stability of the I27 and I28 modules of human cardiac titin. Force-clamp spectroscopy promises to be a direct way to probe the mechanical stability of elastic proteins such as those found in muscle, the extracellular matrix, and cell adhesion.

  19. The TCF C-clamp DNA binding domain expands the Wnt transcriptome via alternative target recognition.

    Science.gov (United States)

    Hoverter, Nate P; Zeller, Michael D; McQuade, Miriam M; Garibaldi, Angela; Busch, Anke; Selwan, Elizabeth M; Hertel, Klemens J; Baldi, Pierre; Waterman, Marian L

    2014-12-16

    LEF/TCFs direct the final step in Wnt/β-catenin signalling by recruiting β-catenin to genes for activation of transcription. Ancient, non-vertebrate TCFs contain two DNA binding domains, a High Mobility Group box for recognition of the Wnt Response Element (WRE; 5'-CTTTGWWS-3') and the C-clamp domain for recognition of the GC-rich Helper motif (5'-RCCGCC-3'). Two vertebrate TCFs (TCF-1/TCF7 and TCF-4/TCF7L2) use the C-clamp as an alternatively spliced domain to regulate cell-cycle progression, but how the C-clamp influences TCF binding and activity genome-wide is not known. Here, we used a doxycycline inducible system with ChIP-seq to assess how the C-clamp influences human TCF1 binding genome-wide. Metabolic pulse-labeling of nascent RNA with 4'Thiouridine was used with RNA-seq to connect binding to the Wnt transcriptome. We find that the C-clamp enables targeting to a greater number of gene loci for stronger occupancy and transcription regulation. The C-clamp uses Helper sites concurrently with WREs for gene targeting, but it also targets TCF1 to sites that do not have readily identifiable canonical WREs. The coupled ChIP-seq/4'Thiouridine-seq analysis identified new Wnt target genes, including additional regulators of cell proliferation. Thus, C-clamp containing isoforms of TCFs are potent transcriptional regulators with an expanded transcriptome directed by C-clamp-Helper site interactions. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  20. Delayed umbilical cord clamping after childbirth: potential benefits to baby's health

    Directory of Open Access Journals (Sweden)

    Uwins C

    2014-11-01

    Full Text Available Christina Uwins,1 David JR Hutchon2 1Department of Obstetrics and Gynaecology, Croydon University Hospital, Croydon, 2Department of Obstetrics, Darlington Memorial Hospital, Darlington, UKAbstract: Early cord clamping was initially introduced as part of the package of care known as “active management of the third stage”, which was implemented to reduce postpartum hemorrhage. It has now been shown to provide no benefit to the mother and to result in harm to the neonate. The clinical trial evidence relating to delayed cord clamping compared to immediate cord clamping is presented and the physiological rationale for delayed cord clamping is discussed in this paper. Most organizations (eg World Health Organization (WHO, Royal College of Obstetricians and Gynaecologists (RCOG, Resuscitation Council (UK,The International Federation of Gynecology and Obstetrics (FIGO, International Confederation of Midwives, International Liaison Committee on Resuscitation (ILCOR and the European Resuscitation Council now advise a delay of 1–3 minutes before clamping the cord in term and preterm infants, and clinicians need to be aware of this change. Healthy neonates benefit from a more physiological and gentle transition from placental to pulmonary respiration, and we explain why this benefit should be provided to all neonates until there is any evidence to the contrary. The harm of early cord clamping is not limited to anemia and iron deficiency, and evidence for a wide range of possible harms of early cord clamping is presented. The need for resuscitation is one of the most common concerns, and ways of overcoming these concerns are described.Keywords: transition, cord clamping, hypovolemia, intraventricular hemorrhage

  1. Patch Clamp: A Powerful Technique for Studying the Mechanism of Acupuncture

    Directory of Open Access Journals (Sweden)

    D. Zhang

    2012-01-01

    Full Text Available Cellular and molecular events can be investigated using electrophysiological techniques. In particular, the patch-clamp method provides detailed information. In addition, the patch-clamp technique has become a powerful method for investigating the mechanisms underlying the effects of acupuncture. In this paper, recent researches on how acupuncture might modulate electrophysiological responses in the central nervous system (CNS and affect peripheral structures are reviewed.

  2. [High-resolution patch-clamp technique based on feedback control of scanning ion conductance microscopy].

    Science.gov (United States)

    Yang, Xi; Liu, Xiao; Zhang, Xiao-Fan; Lu, Hu-Jie; Zhang, Yan-Jun

    2010-06-25

    The ion channels located on the cell fine structures play an important role in the physiological functions of cell membrane. However, it is impossible to achieve precise positioning on the nanometer scale cellular microstructures by conventional patch-clamp technique, due to the 200 nm resolution limit of optical microscope. To solve this problem, we have established a high-resolution patch-clamp technique, which combined commercial scanning ion conductance microscopy (SICM) and patch-clamp recording through a nanopipette probe, based on SICM feedback control. MDCK cells were used as observation object to test the capability of the technique. Firstly, a feedback controlled SICM nanopipette (approximately 150 MOmega) non-contactly scanned over a selected area of living MDCK cells monolayer to obtain high-resolution topographic images of microvilli and tight-junction microstructures on the MDCK cells monolayer. Secondly, the same nanopipette was non-contactly moved and precisely positioned over the microvilli or tight-junction microstructure under SICM feedback control. Finally, the SICM feedback control was switched off, the nanopipette slowly contacted with the cell membrane to get a patch-clamp giga-ohm sealing in the cell-attached patch-clamp configuration, and then performed ion channel recording as a normal patch-clamp electrode. The ion channel recordings showed that ion channels of microvilli microstructure opened at pipette holding potential of -100, -60, -40, 0, +40, +60, +100 mV (n=11). However, the opening of ion channels of tight-junction microstructure was not detected at pipette holding potential of -100, -40, 0, +40, +100 mV (n=9). These results suggest that our high-resolution patch-clamp technique can achieve accurate nanopipette positioning and nanometer scale high-resolution patch-clamp recording, which may provide a powerful tool to study the spatial distribution and functions of ion channel in the nanometer scale microstructures of living

  3. Automated whole-cell patch clamp electrophysiology of neurons in vivo

    OpenAIRE

    Kodandaramaiah, Suhasa B.; Franzesi, Giovanni Talei; Chow, Brian Y.; Boyden, Edward S.; Forest, Craig R.

    2012-01-01

    Whole-cell patch clamp electrophysiology of neurons is a gold standard technique for high-fidelity analysis of the biophysical mechanisms of neural computation and pathology but it requires great skill to perform. We have developed a robot that automatically performs patch clamping in vivo, algorithmically detecting cells by analyzing the temporal sequence of electrode impedance changes. We demonstrate good yield, throughput, and quality of automated intracellular recording in mouse cortex an...

  4. Automated whole-cell patch clamp electrophysiology of neurons in vivo

    Science.gov (United States)

    Kodandaramaiah, Suhasa B.; Franzesi, Giovanni Talei; Chow, Brian Y.; Boyden, Edward S.; Forest, Craig R.

    2012-01-01

    Whole-cell patch clamp electrophysiology of neurons is a gold standard technique for high-fidelity analysis of the biophysical mechanisms of neural computation and pathology but it requires great skill to perform. We have developed a robot that automatically performs patch clamping in vivo, algorithmically detecting cells by analyzing the temporal sequence of electrode impedance changes. We demonstrate good yield, throughput, and quality of automated intracellular recording in mouse cortex and hippocampus. PMID:22561988

  5. Locoregional Anesthesia for Carotid Endarterectomy: Identification of Patients with Intolerance to Cross-Clamping.

    Science.gov (United States)

    Dellaretti, Marcos; de Vasconcelos, Laura T; Dourado, Jules; de Souza, Renata F; Fontoura, Renato R; de Sousa, Atos A

    2016-03-01

    During carotid endarterectomy (CEA), carotid cross-clamping is performed to allow for artery incision and plaque removal. A small subgroup of patients can tolerate carotid occlusion for only a few seconds, if at all, without presenting neurologic deficit. These patients are described as having ''cross-clamp intolerance.'' The purpose of this study was to demonstrate the safety of locoregional anesthesia in identifying patients with cross-clamp intolerance and factors associated with this condition. From August 2008 to May 2010, 115 consecutive patients were submitted to CEA under locoregional anesthesia at the Santa Casa de Belo Horizonte; the procedure was performed by the main author. Patients who showed intolerance to internal carotid artery (ICA) occlusion for intolerance. Among the 115 participating patients, 9.6% (11 patients) showed intolerance to ICA occlusion and developed deficits in intolerance). The factor that was associated with cross-clamp intolerance was the mean degree of contralateral carotid stenosis, which was 57.5% for those who presented intolerance and 27.8% for those who tolerated ICA occlusion. Locoregional anesthesia is a safe method for identifying patients with cross-clamp intolerance. Patients with cross-clamp intolerance present contralateral stenosis that is greater on average than patients who readily tolerate carotid artery occlusion. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. A High Step-Down Interleaved Buck Converter with Active-Clamp Circuits for Wind Turbines

    Directory of Open Access Journals (Sweden)

    Chih-Lung Shen

    2012-12-01

    Full Text Available In this paper, a high step-down interleaved buck coupled-inductor converter (IBCC with active-clamp circuits for wind energy conversion has been studied. In high step-down voltage applications, an IBCC can extend duty ratio and reduce voltage stresses on active switches. In order to reduce switching losses of active switches to improve conversion efficiency, a IBCC with soft-switching techniques is usually required. Compared with passive-clamp circuits, the IBCC with active-clamp circuits have lower switching losses and minimum ringing voltage of the active switches. Thus, the proposed IBCC with active-clamp circuits for wind energy conversion can significantly increase conversion efficiency. Finally, a 240 W prototype of the proposed IBCC with active-clamp circuits was built and implemented. Experimental results have shown that efficiency can reach as high as 91%. The proposed IBCC with active-clamp circuits is presented in high step-down voltage applications to verify the performance and the feasibility for energy conversion of wind turbines.

  7. Early identification of hERG liability in drug discovery programs by automated patch clamp

    Directory of Open Access Journals (Sweden)

    Timm eDanker

    2014-09-01

    Full Text Available Blockade of the cardiac ion channel coded by hERG can lead to cardiac arrhythmia, which has become a major concern in drug discovery and development. Automated electrophysiological patch clamp allows assessment of hERG channel effects early in drug development to aid medicinal chemistry programs and has become routine in pharmaceutical companies. However, a number of potential sources of errors in setting up hERG channel assays by automated patch clamp can lead to misinterpretation of data or false effects being reported. This article describes protocols for automated electrophysiology screening of compound effects on the hERG channel current. Protocol details and the translation of criteria known from manual patch clamp experiments to automated patch clamp experiments to achieve good quality data are emphasized. Typical pitfalls and artifacts that may lead to misinterpretation of data are discussed. While this article focuses on hERG channel recordings using the QPatch (Sophion A/S, Copenhagen, Denmark technology, many of the assay and protocol details given in this article can be transferred for setting up different ion channel assays by automated patch clamp and are similar on other planar patch clamp platforms.

  8. Patch-clamp technique to characterize ion channels in enlarged individual endolysosomes.

    Science.gov (United States)

    Chen, Cheng-Chang; Cang, Chunlei; Fenske, Stefanie; Butz, Elisabeth; Chao, Yu-Kai; Biel, Martin; Ren, Dejian; Wahl-Schott, Christian; Grimm, Christian

    2017-08-01

    According to proteomics analyses, more than 70 different ion channels and transporters are harbored in membranes of intracellular compartments such as endosomes and lysosomes. Malfunctioning of these channels has been implicated in human diseases such as lysosomal storage disorders, neurodegenerative diseases and metabolic pathologies, as well as in the progression of certain infectious diseases. As a consequence, these channels have engendered very high interest as future drug targets. Detailed electrophysiological characterization of intracellular ion channels is lacking, mainly because standard methods to analyze plasma membrane ion channels, such as the patch-clamp technique, are not readily applicable to intracellular organelles. Here we present a protocol detailing how to implement a manual patch-clamp technique for endolysosomal compartments. In contrast to the alternatively used planar endolysosomal patch-clamp technique, this method is a visually controlled, direct patch-clamp technique similar to conventional patch-clamping. The protocol assumes basic knowledge and experience with patch-clamp methods. Implementation of the method requires up to 1 week, and material preparation takes ∼2-4 d. An individual experiment (i.e., measurement of channel currents across the endolysosomal membrane), including control experiments, can be completed within 1 h. This excludes the time for endolysosome enlargement, which takes between 1 and 48 h, depending on the approach and cell type used. Data analysis requires an additional hour.

  9. Early identification of hERG liability in drug discovery programs by automated patch clamp.

    Science.gov (United States)

    Danker, Timm; Möller, Clemens

    2014-01-01

    Blockade of the cardiac ion channel coded by human ether-à-gogo-related gene (hERG) can lead to cardiac arrhythmia, which has become a major concern in drug discovery and development. Automated electrophysiological patch clamp allows assessment of hERG channel effects early in drug development to aid medicinal chemistry programs and has become routine in pharmaceutical companies. However, a number of potential sources of errors in setting up hERG channel assays by automated patch clamp can lead to misinterpretation of data or false effects being reported. This article describes protocols for automated electrophysiology screening of compound effects on the hERG channel current. Protocol details and the translation of criteria known from manual patch clamp experiments to automated patch clamp experiments to achieve good quality data are emphasized. Typical pitfalls and artifacts that may lead to misinterpretation of data are discussed. While this article focuses on hERG channel recordings using the QPatch (Sophion A/S, Copenhagen, Denmark) technology, many of the assay and protocol details given in this article can be transferred for setting up different ion channel assays by automated patch clamp and are similar on other planar patch clamp platforms.

  10. The development of high quality seals for silicon patch-clamp chips.

    Science.gov (United States)

    Sordel, Thomas; Kermarrec, Frédérique; Sinquin, Yann; Fonteille, Isabelle; Labeau, Michel; Sauter-Starace, Fabien; Pudda, Catherine; de Crécy, François; Chatelain, François; De Waard, Michel; Arnoult, Christophe; Picollet-D'hahan, Nathalie

    2010-10-01

    Planar patch-clamp is a two-dimensional variation of traditional patch-clamp. By contrast to classical glass micropipette, the seal quality of silicon patch-clamp chips (i.e. seal resistance and seal success rate) have remained poor due to the planar geometry and the nature of the substrate and thus partially obliterate the advantages related to planar patch-clamp. The characterization of physical parameters involved in seal formation is thus of major interest. In this paper, we demonstrate that the physical characterization of surfaces by a set of techniques (Atomic Force Microscopy (AFM), Scanning Electron Microscopy (SEM), X-ray Photoelectron Spectroscopy (XPS), surface energy (polar and dispersive contributions), drop angles, impedance spectroscopy, combined with a statistical design of experiments (DOE)) allowed us discriminating chips that provide relevant performances for planar patch-clamp analysis. Analyses of seal quality demonstrate that dispersive interactions and micropore size are the most crucial physical parameters of chip surfaces, by contrast to surface roughness and dielectric membrane thickness. This multi-scale study combined with electrophysiological validation of chips on a diverse set of cell-types expressing various ion channels (IRK1, hERG and hNa(v)1.5 channels) unveiled a suitable patch-clamp chip candidate. This original approach may inspire novel strategies for selecting appropriate surface parameters dedicated to biochips.

  11. Late umbilical cord clamping, neonatal hematocrit and Apgar scores: a randomized controlled trial.

    Science.gov (United States)

    Salari, Z; Rezapour, M; Khalili, N

    2014-01-01

    Based on current evidence, there is a little agreement on the best timing for after birth umbilical cord clamping. This study was designed to compare the impact of using two different times for cord clamping on hematocrit concentration and Apgar scores of the neonate. Fifty-six healthy full-term vaginally born neonates were allocated to early (10 seconds after delivery) and late (3 minutes after delivery) umbilical cord clamping groups in this randomized clinical trial. We recorded the length of the 3rd stage of labor and Apgar score at 5 minutes. Infant's hematocrit was measured at 2 and 18 hours of age. Neonatal hematocrit differed between the two groups. Late cord clamping group had greater hematocrit at 2 hours (45.5 ± 4 vs. 49.5 ± 4.4, P = 0.0003) and 18 hours (47.7 ± 5.5 vs. 52.9 ± 4.3, P = 0.0002). Apgar scores at 5 minutes (9.3 ± 0.6 vs. 9.4 ± 0.6, p = 0.5) and duration of delivery 3rd stage (10.2 ± 3.7 min vs. 8.9 ± 5 min, P = 0.2) did not differ between early and late cord clamping groups respectively. Late cord clamping leads to a significant increase in the hematocrit of the neonate but it does not have effects on Apgar score and duration of the 3rd stage of labor.

  12. Modeling CICR in rat ventricular myocytes: voltage clamp studies

    Directory of Open Access Journals (Sweden)

    Palade Philip T

    2010-11-01

    Full Text Available Abstract Background The past thirty-five years have seen an intense search for the molecular mechanisms underlying calcium-induced calcium-release (CICR in cardiac myocytes, with voltage clamp (VC studies being the leading tool employed. Several VC protocols including lowering of extracellular calcium to affect Ca2+ loading of the sarcoplasmic reticulum (SR, and administration of blockers caffeine and thapsigargin have been utilized to probe the phenomena surrounding SR Ca2+ release. Here, we develop a deterministic mathematical model of a rat ventricular myocyte under VC conditions, to better understand mechanisms underlying the response of an isolated cell to calcium perturbation. Motivation for the study was to pinpoint key control variables influencing CICR and examine the role of CICR in the context of a physiological control system regulating cytosolic Ca2+ concentration ([Ca2+]myo. Methods The cell model consists of an electrical-equivalent model for the cell membrane and a fluid-compartment model describing the flux of ionic species between the extracellular and several intracellular compartments (cell cytosol, SR and the dyadic coupling unit (DCU, in which resides the mechanistic basis of CICR. The DCU is described as a controller-actuator mechanism, internally stabilized by negative feedback control of the unit's two diametrically-opposed Ca2+ channels (trigger-channel and release-channel. It releases Ca2+ flux into the cyto-plasm and is in turn enclosed within a negative feedback loop involving the SERCA pump, regulating[Ca2+]myo. Results Our model reproduces measured VC data published by several laboratories, and generates graded Ca2+ release at high Ca2+ gain in a homeostatically-controlled environment where [Ca2+]myo is precisely regulated. We elucidate the importance of the DCU elements in this process, particularly the role of the ryanodine receptor in controlling SR Ca2+ release, its activation by trigger Ca2+, and its

  13. The Formation of the Surface during AN Abrasive Finishing at Constant and Variable Clamping Forces: Finishing at Constant and Variable Clamping Forces

    Science.gov (United States)

    Muratov, Karim Ravilevich; Muratov, Ravil Arifovich; Ablyaz, Timur Rizovich; Gashev, Evgeniy Anatolyevich

    2017-06-01

    In this work the flat abrasive finishing process is reported. The mechanical and chemical phenomena taking place during the finishing process are described. The most common forming process schemes observed during abrasive treatment are reviewed. The schemes of pulsed, geometrical and energetic models are discussed. On the basis of the energetic hypothesis and Preston hypothesis, the rational law of clamping force variation during the finishing is established. This law allowed for the stabilization of the contact pressure and an increase of the treatment efficiency. An exponential dependence of a boundary contact area variation during the finishing of planes with different initial profiles of the macro-topography is developed. An experimental confirmation of the Preston hypothesis during the abrasive finishing is carried out with the plane-finishing machine “Rastr 220”. The comparison experiments are carried out at constant and variable clamping forces of the treated surface towards the tool. It is found that by varying the forces according to the exponential law (i.e. similar to the changing of the boundary contact area during the abrasive finishing) the process efficiency is increasing by a factor of 2.5-3. It is shown that the changes of the root mean square deviation σ of the roughness Ra of a treated surface in the beginning of the process (at time t=1 min) during the finishing at a constant clamping force is two times higher compared to the treatment at a variable clamping force in the same time period.

  14. Improvement in cardioplegic perfusion technique in single aortic clamping - initial results

    Science.gov (United States)

    Sobral, Marcelo Luiz Peixoto; dos Santos Júnior, Sérgio Francisco; de Sá, Juliano Cavalcante; Terrazas, Anderson da Silva; Trompieri, Daniel Francisco de Mendonça; de Sousa, Thierry Araújo Nunes; dos Santos, Gilmar Geraldo; Stolf, Noedir Antonio Groppo

    2014-01-01

    Introduction The most common method used for myocardial protection is administering cardioplegic solution in the coronary circulation. Nevertheless, protection may be achieved by intermittent perfusion of the coronary system with patient's own blood. The intermittent perfusion may be performed by multiple sequences of clamping and opening of the aortic clamp or due single clamping and accessory cannulation of the aortic root as in the improved technique proposed in this study, reperfusion without the need for multiple clamping of the aorta. Objective To evaluate the clinical outcome and the occurrence of neurological events in in-hospital patients submitted to myocardial revascularization surgery with the "improved technique" of intermittent perfusion of the aortic root with single clamping. Methods This is a prospective, cross-sectional, observational study that describes a myocardial management technique that consists of intermittent perfusion of the aortic root with single clamping in which 50 patients (mean age 58.5±7.19 years old) have been submitted to the myocardial revasculrization surgery under the proposed technique. Clinical and laboratory variables, pre- and post-surgery, have been assessed. Results The mean peak level of post-surgery CKMB was 51.64±27.10 U/L in the second post-surgery and of troponin I was 3.35±4.39 ng/ml in the fourth post-surgery, within normal limits. No deaths have occurred and one patient presented mild neurological disorder. Hemodynamic monitoring has not indicated any changes. Conclusion The myocardial revascularization surgery by perfusion with the improved technique with intermittent aortic root with single clamping proved to be safe, enabling satisfactory clinical results. PMID:25140473

  15. Pre-Descemets endothelial keratoplasty: the PDEK clamp for successful PDEK.

    Science.gov (United States)

    Dua, H S; Said, D G

    2017-07-01

    PurposeTo design and produce a clamp that enables good handling of donor sclera-corneal disks, allows air to be injected in the corneal stroma and consistently provides pre-Descemets endothelial keratoplasty (PDEK) tissue by creation of a type-1 big bubble (BB) avoiding a type-2BB by shutting the fenestrations in the periphery of the pre-Descemets layer and preventing escape of air.Patients and methodsA clamp with spring loaded handles attached to two rings of 9 mm diameter and 1 mm width with a side port for insertion of a needle for air injection was designed and produced. The clamp was tested in 20 human donor sclera-corneal disks and refined over three prototypes. The final design was used for PDEK in 8 patients.ResultsThe clamp was able to prevent any escape of air or the formation of a type-2BB. By preventing air leak, it provided better control over the volume of air injected allowing for consistent formation of a type-1BB, which could be expanded to its full diameter by advancing the tip of the needle into the bubble cavity. Centration of tissue in the clamp was important. The clamp was used successfully to obtain PDEK tissue from 8 donor sclara-corneal disks for transplant in three eyes with pseudophakic bullous keratopathy and five eyes with pseudophakia and Fuchs' endothelial dystrophy.ConclusionThe PDEK clamp is a simple and useful instrument that will give surgeons the confidence of consistently obtaining PDEK tissue with ease and without the risk of separating the Descemets membrane in a type-2BB.

  16. Conformational Changes of the Clamp of the Protein Translocation ATPase SecA.

    Science.gov (United States)

    Chen, Yu; Bauer, Benedikt W; Rapoport, Tom A; Gumbart, James C

    2015-07-17

    Post-translational protein translocation across the bacterial plasma membrane is mediated by the interplay of the SecA ATPase and the protein-conducting SecY channel. SecA consists of several domains, including two nucleotide-binding domains (NBD1 and NBD2), a polypeptide cross-linking domain (PPXD), a helical scaffold domain (HSD), and a helical wing domain (HWD). PPXD, HSD, and NBD2 form a clamp that positions the polypeptide substrate above the channel so that it can be pushed into the channel by a two-helix finger of the HSD. How the substrate is accommodated in the clamp during translocation is unclear. Here, we report a crystal structure of Thermotoga maritima SecA at 1.9 Å resolution. Structural analysis and free-energy calculations indicate that the new structure represents an intermediate state during the transition of the clamp from an open to a closed conformation. Molecular dynamics simulations show that closure of the clamp occurs in two phases, an initial movement of PPXD, HSD, and HWD as a unit, followed by a movement of PPXD alone toward NBD2. Simulations in the presence of a polypeptide chain show that the substrate associates with the back of the clamp by dynamic hydrogen bonding and that the clamp is laterally closed by a conserved loop of the PPXD. Mutational disruption of clamp opening or closure abolishes protein translocation. These results suggest how conformational changes of SecA allow substrate binding and movement during protein translocation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Clampe de Ganz no tratamento de urgência em lesões do anel pélvico Ganz clamps to treat emergencies in pelvic ring lesions

    Directory of Open Access Journals (Sweden)

    Gilberto José Cação Pereira

    2008-07-01

    Full Text Available OBJETIVO: Avaliar a eficiência do clampe de Ganz na estabilização e redução dos deslocamentos da pelve, quando utilizado no tratamento de urgência, além de aquilatar eventuais dificuldades e facilidades do método. MÉTODOS: O clampe de Ganz foi utilizado no tratamento de urgência em 31 pacientes com graves lesões do anel pélvico (Tile C associadas a importante instabilidade hemodinâmica. RESULTADO: Entre os pacientes, 27 (87,1% apresentaram evolução favorável, com estabilização, redução dos deslocamentos e compressão da região posterior do anel, além de estabilização hemodinâmica, e quatro (12,9% evoluíram para óbito. CONCLUSÃO: O clampe de Ganz mostrou-se eficiente ferramenta no tratamento de urgência das lesões do anel pélvico, por ser de concepção simples, de rápida colocação, não impedir ou dificultar procedimentos no abdome e, principalmente, por permitir a estabilização da pelve, redução dos deslocamentos e compressão na região posterior do anel, local onde ocorrem os maiores sangramentos, reduzindo-os ou eliminando-os.OBJECTIVE: To evaluate the effectiveness of Ganz clamp to stabilize and reduce pelvic dislocations in emergency situations, and to assess possible difficult and easy aspects of the method. METHOD: Ganz clamp was used in the emergency treatment of 31 patients with severe lesions of the pelvic ring (Tile C associated to major hemodynamic instability. RESULTS: Among the patients, 27 (87.1% presented a favorable evolution, with stabilization, reduction of dislocations, and compression of the posterior portion of the ring, and also hemodynamic stabilization, and 4 (12.9% died. CONCLUSION: The Ganz clamp was found to be an effective tool in the emergency treatment of pelvic ring lesions because it has an uncomplicated conception, allowing for a fast placement, because it does not prevent nor make it difficult for abdominal procedures, and most of all because it allows for the

  18. Functional dissection of synaptic circuits: in vivo patch-clamp recording in neuroscience.

    Science.gov (United States)

    Tao, Can; Zhang, Guangwei; Xiong, Ying; Zhou, Yi

    2015-01-01

    Neuronal activity is dominated by synaptic inputs from excitatory or inhibitory neural circuits. With the development of in vivo patch-clamp recording, especially in vivo voltage-clamp recording, researchers can not only directly measure neuronal activity, such as spiking responses or membrane potential dynamics, but also quantify synaptic inputs from excitatory and inhibitory circuits in living animals. This approach enables researchers to directly unravel different synaptic components and to understand their underlying roles in particular brain functions. Combining in vivo patch-clamp recording with other techniques, such as two-photon imaging or optogenetics, can provide even clearer functional dissection of the synaptic contributions of different neurons or nuclei. Here, we summarized current applications and recent research progress using the in vivo patch-clamp recording method and focused on its role in the functional dissection of different synaptic inputs. The key factors of a successful in vivo patch-clamp experiment and possible solutions based on references and our experiences were also discussed.

  19. A single electrode voltage, current- and patch-clamp amplifier with complete stable series resistance compensation.

    Science.gov (United States)

    Strickholm, A

    1995-01-01

    An input headstage for single electrode voltage and current clamping is described which permits selecting desired operational modes during an experiment. These include cell attached or whole cell patch clamping, single micro-electrode voltage or current clamping, potential recording, iontophoresis and voltammetry. Input electrode series resistance with the electrode inserted in a cell or during whole cell patch clamping can be measured at any time. The unique circuitry allows complete compensation of the series resistance with high frequency response. The input series electrode resistance can also vary considerably around an initially set 100% compensation with intrinsic stability. An accelerator 'supercharger' or 'booster circuit' is included which can shorten the rise time of membrane potential to the command potential by a hundredfold. This improves the recording of early ion channel currents. Feedback resistors are selected remotely for the specific experimental need. Signal resolution in patch clamping is that expected from the thermal noise of the feedback resistor and electrode to membrane seal resistance. This headstage circuitry design thus allows interchangeable modes of operation and parameters during an experiment to obtain optimal conditions for signal detection and frequency response.

  20. Clamping instability and van der Waals forces in carbon nanotube mechanical resonators.

    Science.gov (United States)

    Aykol, Mehmet; Hou, Bingya; Dhall, Rohan; Chang, Shun-Wen; Branham, William; Qiu, Jing; Cronin, Stephen B

    2014-05-14

    We investigate the role of weak clamping forces, typically assumed to be infinite, in carbon nanotube mechanical resonators. Due to these forces, we observe a hysteretic clamping and unclamping of the nanotube device that results in a discrete drop in the mechanical resonance frequency on the order of 5-20 MHz, when the temperature is cycled between 340 and 375 K. This instability in the resonant frequency results from the nanotube unpinning from the electrode/trench sidewall where it is bound weakly by van der Waals forces. Interestingly, this unpinning does not affect the Q-factor of the resonance, since the clamping is still governed by van der Waals forces above and below the unpinning. For a 1 μm device, the drop observed in resonance frequency corresponds to a change in nanotube length of approximately 50-65 nm. On the basis of these findings, we introduce a new model, which includes a finite tension around zero gate voltage due to van der Waals forces and shows better agreement with the experimental data than the perfect clamping model. From the gate dependence of the mechanical resonance frequency, we extract the van der Waals clamping force to be 1.8 pN. The mechanical resonance frequency exhibits a striking temperature dependence below 200 K attributed to a temperature-dependent slack arising from the competition between the van der Waals force and the thermal fluctuations in the suspended nanotube.

  1. Combined Triplex/Duplex Invasion of Double-Stranded DNA by "Tail-Clamp" Peptide Nucleic Acid

    DEFF Research Database (Denmark)

    Bentin, Thomas; Larsen, H. J.; Nielsen, Peter E.

    2003-01-01

    as determined by T-m measurements. Binding to double-stranded (ds) DNA occurred by combined triplex and duplex invasion as analyzed by permanganate probing. Furthermore, C-50 measurements revealed that tail-clamp PNAs consistently bound the dsDNA target more efficiently, and kinetics experiments revealed......"Tail-clamp" PNAs composed of a short (hexamer) homopyrimidine triplex forming domain and a (decamer) mixed sequence duplex forming extension have been designed. Tail-clamp PNAs display significantly increased binding to single-stranded DNA compared with PNAs lacking a duplex-forming extension...... that this was due to a dramatically reduced dissociation rate of such complexes. Increasing the PNA net charge also increased binding efficiency, but unexpectedly, this increase was much more pronounced for tailless-clamp PNAs than for tail-clamp PNAs. Finally, shortening the tail-clamp PNA triplex invasion moiety...

  2. The role of acute hyperinsulinemia in the development of cardiac arrhythmias.

    Science.gov (United States)

    Drimba, László; Döbrönte, Róbert; Hegedüs, Csaba; Sári, Réka; Di, Yin; Németh, Joseph; Szilvássy, Zoltán; Peitl, Barna

    2013-05-01

    Patients with perturbed metabolic control are more prone to develop cardiac rhythm disturbances. The main purpose of the present preclinical study was to investigate the possible role of euglycemic hyperinsulinemia in development of cardiac arrhythmias. Euglycemic hyperinsulinemia was induced in conscious rabbits equipped with a right ventricular pacemaker electrode catheter by hyperinsulinemic euglycemic glucose clamp (HEGC) applying two different rates of insulin infusion (5 and 10 mIU/kg/min) and variable rate of glucose infusion to maintain euglycemia (5.5 ± 0.5 mmol/l). The effect of hyperinsulinemia on cardiac electrophysiological parameters was continuously monitored by means of 12-lead surface ECG recording. Arrhythmia incidence was determined by means of programmed electrical stimulation (PES). The possible role of adrenergic activation was investigated by determination of plasma catecholamine levels and intravenous administration of a beta adrenergic blocking agent, metoprolol. All of the measurements were performed during the steady-state period of HEGC and subsequent to metoprolol administration. Both 5 and 10 mIU/kg/min insulin infusion prolonged significantly QTend, QTc, and Tpeak-Tend intervals. The incidence of ventricular arrhythmias generated by PES was increased significantly by euglycemic hyperinsulinemia and exhibited linear relationship to plasma levels of insulin. No alteration on plasma catecholamine levels could be observed; however, metoprolol treatment restored the prolonged QTend, QTc, and Tpeak-Tend intervals and significantly reduced the hyperinsulinemia-induced increase of arrhythmia incidence. Euglycemic hyperinsulinemia can exert proarrhythmic effect presumably due to the enhancement of transmural dispersion of repolarization. Metoprolol treatment may be of benefit in hyperinsulinemia associated with increased incidence of cardiac arrhythmias.

  3. Leucine disposal rate for assessment of amino acid metabolism in maintenance hemodialysis patients

    Science.gov (United States)

    Denny, Gerald B.; Deger, Serpil M.; Chen, Guanhua; Bian, Aihua; Sha, Feng; Booker, Cindy; Kesler, Jaclyn T.; David, Sthuthi; Ellis, Charles D.; Ikizler, T. Alp

    2016-01-01

    Background Protein energy wasting (PEW) is common in patients undergoing maintenance hemodialysis (MHD) and closely associated with poor outcomes. Insulin resistance and associated alterations in amino acid metabolism are potential pathways leading to PEW. We hypothesized that the measurement of leucine disposal during a hyperinsulinemic- euglycemic-euaminoacidemic clamp (HEAC) procedure would accurately measure the sensitivity to insulin for its actions on concomitant carbohydrate and protein metabolism in MHD patients. Methods We examined 35 MHD patients and 17 control subjects with normal kidney function by hyperinsulinemic-euglycemic clamp (HEGC) followed by HEAC clamp procedure to obtain leucine disposal rate (LDR) along with isotope tracer methodology to assess whole body protein turnover. Results The glucose disposal rate (GDR) by HEGC was 5.1 ± 2.1 mg/kg/min for the MHD patients compared to 6.3 ± 3.9 mg/kg/min for the controls (p = 0.38). The LDR during HEAC was 0.09 ± 0.03 mg/kg/min for the MHD patients compared to 0.11 ± 0.05 mg/kg/min for the controls (p = 0.009). The LDR level was correlated with whole body protein synthesis (r = 0.25; p = 0.08), with whole body protein breakdown (r = −0.38 p = 0.01) and net protein balance (r = 0.85; p < 0.001) in the overall study population. Correlations remained significant in subgroup analysis. The GDR derived by HEGC and LDR correlated well in the controls (r = 0.79, p < 0.001), but less so in the MHD patients (r = 0.58, p < 0.001). Conclusions Leucine disposal rate reliably measures amino acid utilization in MHD patients and controls in response to high dose insulin. PMID:27413537

  4. No effect on the short-term of a decrease in blood viscosity on insulin resistance.

    Science.gov (United States)

    Gnasso, Agostino; Cacia, Michele; Cortese, Claudio; Succurro, Elena; Andreozzi, Francesco; Carallo, Claudio; Irace, Concetta

    2018-01-01

    Blood viscosity (BV) might influence glucose delivery to peripheral tissues and play an important role in insulin resistance and diabetes mellitus. However, the exact relationship between BV and insulin resistance is not yet clear. Aim of the present study is to evaluate the effects of the acute reduction in BV on insulin resistance, in healthy male subjects. Fifteen healthy male blood donors have been studied before and 48 hours after blood donation. Blood and plasma viscosity have been measured at 37°C with a cone-plate viscometer. Insulin resistance has been evaluated by euglycemic/hyperinsulinemic clamp in eight subjects, and by iHOMA2 Index in further seven subjects. Blood viscosity was markedly reduced after blood donation (BV225 (cP) 4.53 ± 0.59 vs. 4.18 ± 0.31, p < 0.05). Insulin resistance was unchanged: MFFM clamp: 5.6 ± 4.5vs. 4.4 ± 2.2 and iHOMA2 Index 1.2 ± 0.6 vs. 1.2 ± 0.5, before vs. after respectively, p = NS. Blood pressure and lipids were unchanged after blood donation. The present results demonstrate that acute reduction of BV in healthy male subjects does not change the insulin resistance, measured using both euglycemic/hyperinsulinemic clamp and iHOMA2 Index. Further intervention studies are needed to assess the effect that the reduction in BV can have in subjects with insulin resistance.

  5. The interplay of primer-template DNA phosphorylation status and single-stranded DNA binding proteins in directing clamp loaders to the appropriate polarity of DNA.

    Science.gov (United States)

    Hayner, Jaclyn N; Douma, Lauren G; Bloom, Linda B

    2014-01-01

    Sliding clamps are loaded onto DNA by clamp loaders to serve the critical role of coordinating various enzymes on DNA. Clamp loaders must quickly and efficiently load clamps at primer/template (p/t) junctions containing a duplex region with a free 3'OH (3'DNA), but it is unclear how clamp loaders target these sites. To measure the Escherichia coli and Saccharomyces cerevisiae clamp loader specificity toward 3'DNA, fluorescent β and PCNA clamps were used to measure clamp closing triggered by DNA substrates of differing polarity, testing the role of both the 5'phosphate (5'P) and the presence of single-stranded binding proteins (SSBs). SSBs inhibit clamp loading by both clamp loaders on the incorrect polarity of DNA (5'DNA). The 5'P groups contribute selectivity to differing degrees for the two clamp loaders, suggesting variations in the mechanism by which clamp loaders target 3'DNA. Interestingly, the χ subunit of the E. coli clamp loader is not required for SSB to inhibit clamp loading on phosphorylated 5'DNA, showing that χ·SSB interactions are dispensable. These studies highlight a common role for SSBs in directing clamp loaders to 3'DNA, as well as uncover nuances in the mechanisms by which SSBs perform this vital role. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  6. Ion channels in small cells and subcellular structures can be studied with a smart patch-clamp system.

    OpenAIRE

    Gorelik, Julia; Gu, Yuchun; Spohr, Hilmar A; Shevchuk, Andrew I; Lab, Max J; Harding, Sian E; Edwards, Christopher R W; Whitaker, Michael; Moss, Guy W J; Benton, David C H; Sánchez, Daniel; Darszon, Alberto; Vodyanoy, Igor; Klenerman, David; Korchev, Yuri E

    2002-01-01

    We have developed a scanning patch-clamp technique that facilitates single-channel recording from small cells and submicron cellular structures that are inaccessible by conventional methods. The scanning patch-clamp technique combines scanning ion conductance microscopy and patch-clamp recording through a single glass nanopipette probe. In this method the nanopipette is first scanned over a cell surface, using current feedback, to obtain a high-resolution topographic image. This same pipette ...

  7. The Effect of an Added Mass on the Frequency Shifts of a Clamped-Clamped Microbeam for Bio-Mass Detection

    KAUST Repository

    Bouchaala, Adam M.

    2016-12-05

    We present analytical formulations to calculate the induced resonance frequency shifts of electrically actuated clamped-clamped microbeams due to an added mass. Based on the Euler-Bernoulli beam theory, we investigate the linear dynamic responses of the beams added masses, which are modeled as discrete point masses. Analytical expressions based on perturbation techniques and a one-mode Galerkin approximation are developed to calculate accurately the frequency shifts under a DC voltage as a function of the added mass and position. The analytical results are compared to numerical solution of the eigenvalue problem. Results are shown for the fundamental as well as the higher-order modes of the beams. The results indicate a significant increase in the frequency shift, and hence the sensitivity of detection, when scaling down to nano scale and using higher-order modes.

  8. Modelling the clamping force distribution among chips in press-pack IGBTs using the finite element method

    DEFF Research Database (Denmark)

    Hasmasan, Adrian Augustin; Busca, Christian; Teodorescu, Remus

    2012-01-01

    In this paper, a FEM (finite element method) based mechanical model for PP (press-pack) IGBTs (insulated gate bipolar transistors) is presented, which can be used to calculate the clamping force distribution among chips under various clamping conditions. The clamping force is an important parameter...... for the chip, because it influences contact electrical resistance, contact thermal resistance and power cycling capability. Ideally, the clamping force should be equally distributed among chips, in order to maximize the reliability of the PP IGBT. The model is built around a hypothetical PP IGBT with 9 chips...

  9. The essential Drosophila CLAMP protein differentially regulates non-coding roX RNAs in male and females.

    Science.gov (United States)

    Urban, Jennifer A; Doherty, Caroline A; Jordan, William T; Bliss, Jacob E; Feng, Jessica; Soruco, Marcela M; Rieder, Leila E; Tsiarli, Maria A; Larschan, Erica N

    2017-06-01

    Heterogametic species require chromosome-wide gene regulation to compensate for differences in sex chromosome gene dosage. In Drosophila melanogaster, transcriptional output from the single male X-chromosome is equalized to that of XX females by recruitment of the male-specific lethal (MSL) complex, which increases transcript levels of active genes 2-fold. The MSL complex contains several protein components and two non-coding RNA on the X ( roX) RNAs that are transcriptionally activated by the MSL complex. We previously discovered that targeting of the MSL complex to the X-chromosome is dependent on the chromatin-linked adapter for MSL proteins (CLAMP) zinc finger protein. To better understand CLAMP function, we used the CRISPR/Cas9 genome editing system to generate a frameshift mutation in the clamp gene that eliminates expression of the CLAMP protein. We found that clamp null females die at the third instar larval stage, while almost all clamp null males die at earlier developmental stages. Moreover, we found that in clamp null females roX gene expression is activated, whereas in clamp null males roX gene expression is reduced. Therefore, CLAMP regulates roX abundance in a sex-specific manner. Our results provide new insights into sex-specific gene regulation by an essential transcription factor.

  10. Significance of Shrinkage Induced Clamping Pressure in Fiber-Matrix Bonding in Cementitious Composite Materials

    DEFF Research Database (Denmark)

    Stang, Henrik

    1996-01-01

    inhomogeneity embedded in a matrix consisting of acementitious material undergoing shrinkage during hydration(autogenous shrinkage). Furthermore, the paperpresents the analysis necessary to perform an interpretation of the experimental results and which allows for thedetermination of the clamping pressure......The present paper accesses the significance of shrinkage inducedclamping pressure in fiber/matrix bonding mechanisms incementitious composite materials. The paper contains a description of an experimental setup whichallows mbox{measurement} of the clamping pressure which develops on anelastic...... acting on any elastic inhomogeneityembedded in the same cementitious matrix material. Fiber-shaped inhomogeneities are of special interest in cementitious composite material systems andresults are presented for the development of clamping pressure on three typical fiber types in two typical cementpastes...

  11. Flange joint system for SRF cavities utilizing high force spring clamps for low particle generation

    Energy Technology Data Exchange (ETDEWEB)

    None

    2017-09-05

    A flange joint system for SRF cavities. The flange joint system includes a set of high force spring clamps that produce high force on the simple flanges of Superconducting Radio Frequency (SRF) cavities to squeeze conventional metallic seals. The system establishes the required vacuum and RF-tight seal with minimum particle contamination to the inside of the cavity assembly. The spring clamps are designed to stay within their elastic range while being forced open enough to mount over the flange pair. Upon release, the clamps have enough force to plastically deform metallic seal surfaces and continue to a new equilibrium sprung dimension where the flanges remain held against one another with enough preload such that normal handling will not break the seal.

  12. Development of Automated Patch Clamp Technique to Investigate CFTR Chloride Channel Function.

    Science.gov (United States)

    Billet, Arnaud; Froux, Lionel; Hanrahan, John W; Becq, Frederic

    2017-01-01

    The chloride (Cl-) channel cystic fibrosis transmembrane conductance regulator (CFTR) is defective in cystic fibrosis (CF), and mutation of its encoding gene leads to various defects such as retention of the misfolded protein in the endoplasmic reticulum, reduced stability at the plasma membrane, abnormal channel gating with low open probability, and thermal instability, which leads to inactivation of the channel at physiological temperature. Pharmacotherapy is one major therapeutic approach in the CF field and needs sensible and fast tools to identify promising compounds. The high throughput screening assays available are often fast and sensible techniques but with lack of specificity. Few works used automated patch clamp (APC) for CFTR recording, and none have compared conventional and planar techniques and demonstrated their capabilities for different types of experiments. In this study, we evaluated the use of planar parallel APC technique for pharmacological search of CFTR-trafficking correctors and CFTR function modulators. Using optimized conditions, we recorded both wt- and corrected F508del-CFTR Cl- currents with automated whole-cell patch clamp and compared the data to results obtained with conventional manual whole-cell patch clamp. We found no significant difference in patch clamp parameters such as cell capacitance and series resistance between automated and manual patch clamp. Also, the results showed good similarities of CFTR currents recording between the two methods. We showed that similar stimulation protocols could be used in both manual and automatic techniques allowing precise control of temperature, classic I/V relationship, and monitoring of current stability in time. In conclusion, parallel patch-clamp recording allows rapid and efficient investigation of CFTR currents with a variety of tests available and could be considered as new tool for medium throughput screening in CF pharmacotherapy.

  13. The alcohol clamp: applications, challenges, and new directions--an RSA 2004 symposium summary.

    Science.gov (United States)

    Ramchandani, Vijay A; O'connor, Sean; Neumark, Yehuda; Zimmermann, Ulrich S; Morzorati, Sandra L; de Wit, Harriet

    2006-01-01

    This article summarizes the proceedings of a symposium organized and cochaired by Vijay Ramchandani and Sean O'Connor and presented at the 2004 Research Society on Alcoholism meeting in Vancouver, BC, Canada. The objectives of this symposium were: (1) to provide a rationale for the development and use of the alcohol clamp and the requirements for its use in alcohol challenge studies; (2) to highlight recent studies conducted using the alcohol clamp to identify sources of variation in the pharmacokinetics and pharmacodynamics of alcohol, as well as to address important research questions related to the relationship between the response to alcohol and the risk for alcoholism; and (3) to provide a perspective on progress, address limitations of the clamp, and identify new directions for alcohol challenge research. The symposium began with an introduction and overview of the alcohol clamp, by Vijay Ramchandani. This was followed by 4 presentations that highlighted recent studies conducted using the clamp including: (1) determination of the influence of alcohol dehydrogenase polymorphisms on alcohol elimination rates in a male Jewish population, by Yehuda Neumark; (2) examination of family history of alcoholism, recent drinking history, and levels and rates of administration as determinants of the response to alcohol and risk for alcoholism, by Sean O'Connor; (3) evaluation of the time course of ethanol intoxication on neuroendocrine function in humans, by Ulrich Zimmermann; and (4) a study of the effects of steady-state blood alcohol levels on auditory event-related potentials in rats, by Sandra Morzorati. Harriet de Wit summarized and discussed the research presented at the symposium and provided her perspective on future directions for research using the alcohol clamp.

  14. Whole cell voltage clamp recordings from cultured neurons of the supraoptic area of neonatal rat hypothalamus.

    Science.gov (United States)

    Cobbett, P; Mason, W T

    1987-04-14

    Whole-cell voltage and current clamp recordings were made from cultured neurons from the hypothalamic supraoptic area of neonatal rats. These neurons fired spontaneous Na+-action potentials, appearing as mixed Na+/K+-currents in voltage clamp. Isolated Na+-currents (less than 3 nA) were rapidly activated and inactivated during positive potential pulses from -80 mV. Two voltage-activated Ba2+-currents (less than 1 nA) were also recorded. These techniques offer a promising new approach for studying the striking electrical behavior of cultured hypothalamic neurons.

  15. Early vs delayed clamping of the umbilical cord in full term, preterm and very preterm infants

    DEFF Research Database (Denmark)

    Moller, N.K.; Weber, T.

    2008-01-01

    Randomized studies from 2006 and two meta-analyses published in 2007 agree that clamping of the umbilical cord can be delayed. For the preterm and very preterm infant benefits include less need for blood transfusion and less morbidity, especially for the very preterm male infant. For the term...

  16. Experimental and numerical analysis of clamped joints in front motorbike suspensions

    Directory of Open Access Journals (Sweden)

    Vincenzi N.

    2010-06-01

    Full Text Available Clamped joints are shaft-hub connections used, as an instance, in front motorbike suspensions to lock the steering plates with the legs and the legs with the wheel pin, by means of one or two bolts. The preloading force, produced during the tightening process, should be evaluated accurately, since it must lock safely the shaft, without overcoming the yielding point of the hub. Firstly, friction coefficients have been evaluated on “ad-hoc designed” specimens, by applying the Design of Experiment approach: the applied tightening torque has been precisely related to the imposed preloading force. Then, the tensile state of clamps have been evaluated both via FEM and by leveraging some design formulae proposed by the Authors as function of the preloading force and of the clamp geometry. Finally, the results have been compared to those given by some strain gauges applied on the tested clamps: the discrepancies between numerical analyses, the design formulae and the experimental results remains under a threshold of 10%.

  17. Fundamentals of a Multi-Phase, Neutral-Point Clamped Multilevel ...

    African Journals Online (AJOL)

    The conventional five-level diode-clamped inverter suffers from the problem of many active power switches when extended to multiphase system. As a consequence, increasing number of gate drive circuits are required; resulting in complex system circuitry. This paper presents the fundamentals of a multiphase, neutral ...

  18. Acute hypotension induced by aortic clamp vs. PTH provokes distinct proximal tubule Na+ transporter redistribution patterns

    DEFF Research Database (Denmark)

    Leong, Patrick K K; Yang, Li E; Lin, Harrison W

    2004-01-01

    in renal cortical membranes fractionated on sorbitol density gradients. Aortic clamp-induced acute hypotension (from 100 +/- 3 to 78 +/- 2 mmHg) provoked a 62% decrease in urine output and a significant decrease in volume flow from the proximal tubule detected as a 66% decrease in endogenous lithium......-density membranes enriched in apical markers. PTH at much lower doses (

  19. Emergent management of APC-2 pelvic ring injuries with an anteriorly placed C-clamp.

    Science.gov (United States)

    Richard, Marc J; Tornetta, Paul

    2009-01-01

    Can anteriorly placed pelvic C-clamps be used successfully in the emergent management of APC-2 pelvic fractures? Prospective cohort. Level 1 trauma center. A single-surgeon series of 24 patients with an anteroposterior compression type 2 pelvic fracture. Application of an anteriorly placed pelvic C-clamp within 2 hours of presentation. Response to hypotension, complications related to pin placement, application time, and symphyseal reduction measured on anteroposterior radiograph. Twenty-four patients with a mean age of 29 years (14-58 years) had an APC-2 pelvic fracture diagnosed by an anteroposterior radiograph of the pelvis on presentation. All patients were emergently managed with an anteriorly placed C-clamp applied in the emergency room (10), angiography suite (9), or operating room (5). Eleven patients presented with hypotension (systolic blood pressure elevation of their blood pressure of 23 mm Hg (10-44 mm Hg). The symphyseal separation was reduced from a mean of 4.5 cm (3-9 cm) to pelvic C-clamp can be placed anteriorly as a part of the early management of APC-2 pelvic fractures with a short application time in a variety of patient care areas.

  20. Cyclic peptide inhibitors of the β-sliding clamp in Staphylococcus aureus

    DEFF Research Database (Denmark)

    Kjelstrup, Susanne; Hansen, Paula Melo Paulon; Thomsen, Line Elnif

    2013-01-01

    , from a library generated by the split intein-mediated circular ligation of peptides and proteins technology, were found to interfere with dimerization of the β-sliding clamp of the replisome. Two 8-mer peptides were analyzed in more detail. Both inhibited DNA replication, led to SOS induction, altered...

  1. Enhanced Buck-Boost Three-Level Neutral-Point-Clamped Inverters

    DEFF Research Database (Denmark)

    Tan, K. K.; Gao, F.; Chiang Loh, Poh

    2009-01-01

    In traditional three-level neutral-point-clamped (NPC) inverters, a major issue is capacitor voltage imbalance, which results in low order harmonics. The compensation of the capacitor voltages often require additional control complexity, which cannot be conveniently implemented. The "alternative...

  2. Electrophysiological analysis of mammalian cells expressing hERG using automated 384-well-patch-clamp.

    Science.gov (United States)

    Haraguchi, Yuji; Ohtsuki, Atsushi; Oka, Takayuki; Shimizu, Tatsuya

    2015-12-16

    An in vitro electrophysiological assay system, which can assess compound effects and thus show cardiotoxicity including arrhythmia risks of test drugs, is an essential method in the field of drug development and toxicology. In this study, high-throughput electrophysiological recordings of human embryonic kidney (HEK 293) cells and Chinese hamster ovary (CHO) cells stably expressing human ether-a-go-go related gene (hERG) were performed utilizing an automated 384-well-patch-clamp system, which records up to 384 cells simultaneously. hERG channel inhibition, which is closely related to a drug-induced QT prolongation and is increasing the risk of sudden cardiac death, was investigated in the high-throughput screening patch-clamp system. In the automated patch-clamp measurements performed here, Kv currents were investigated with high efficiency. Various hERG channel blockers showed concentration-dependent inhibition, the 50 % inhibitory concentrations (IC50) of those blockers were in good agreement with previous reports. The high-throughput patch-clamp system has a high potential in the field of pharmacology, toxicology, and cardiac physiology, and will contribute to the acceleration of pharmaceutical drug development and drug safety testing.

  3. Macromechanical parametric amplification with a base-excited doubly clamped beam

    DEFF Research Database (Denmark)

    Neumeyer, Stefan; Thomsen, Jon Juel

    2013-01-01

    amplitude is derived. For a given small level of transverse displacement for both the cantilever and doubly clamped beam, the effect of midplane stretching is dominant compared to those caused by nonlinear inertia and curvature. It was found that the beam slenderness ratio can be used as an effective design...

  4. Hand-assisted partial nephrectomy with early arterial clamp removal: Impact of the learning curve

    DEFF Research Database (Denmark)

    Azawi, Nessn H; Norus, Thomas P; Wittendorff, Hans-Erik

    2014-01-01

    (40%), nine (45%) and 14 (70%) patients in cohorts 1, 2 and 3, respectively (p = 0.0185). CONCLUSIONS: Hand-assisted laparoscopic partial nephrectomy with early removal of arterial clamps is safe and easy to learn. An expert laparoscopic surgeon can perform hand-assisted laparoscopic partial...

  5. Construction, Calibration, and Validation of a Simple Patch-Clamp Amplifier for Physiology Education

    Science.gov (United States)

    Rouzrokh, Ali; Ebrahimi, Soltan Ahmed; Mahmoudian, Massoud

    2009-01-01

    A modular patch-clamp amplifier was constructed based on the Strickholm design, which was initially published in 1995. Various parts of the amplifier such as the power supply, input circuit, headstage, feedback circuit, output and nulling circuits were redesigned to use recent software advances and fabricated using the common lithographic printed…

  6. A New Preparation of Pancreatic Ducts for Patch-Clamp Studies

    DEFF Research Database (Denmark)

    Novak, Ivana; Hug, M.J.

    1995-01-01

    parameters, monitored with the whole-cell patch-clamp method, and those in isolated perfused ducts, monitored with the conventional microelectrodes [Novak I, Greger R: Pflügers Arch 1988;411:58-68, 546-553], are comparable. Similar conclusion can be made about the calcium measurements [Hug M, et al: Pflügers...

  7. The Response Clamp: Functional characterization of neural systems using closed-loop control

    Directory of Open Access Journals (Sweden)

    Avner eWallach

    2013-01-01

    Full Text Available The voltage clamp method, pioneered by Hodgkin, Huxley and Katz, laid the foundations to neurophysiological research. Its core rationale is the use of closed-loop control as a tool for system characterization. A recently introduced method, the response clamp, extends the voltage clamp rationale to the functional, phenomenological level. The method consists of on-line estimation of a response variable of interest (e.g. the probability of response or its latency and a simple feedback control mechanism designed to tightly converge this variable towards a desired trajectory. In the present contribution I offer a perspective on this novel method and its applications in the broader context of system identification and characterization. First, I demonstrate how internal state variables are exposed using the method, and how the use of several controllers may allow for a detailed, multi-variable characterization of the system. Second, I discuss three different categories of applications of the method: (i exploration of intrinsically generated dynamics, (ii exploration of extrinsically generated dynamics and (iii generation of input-output trajectories. The relation of these categories to similar uses in the voltage clamp and other techniques is also discussed. Finally, I discuss the method’s limitations, as well as its possible synthesis with existing complementary approaches.

  8. Delayed umbilical cord clamping for reducing anaemia in low birthweight infants : implications for developing countries

    NARCIS (Netherlands)

    van Rheenen, Patrick F.; Gruschke, Sebastian; Brabin, Bernard J.

    BACKGROUND: Cheap and effective interventions are needed to reduce the risk of infant anaemia in developing countries. Delayed cord clamping (DCC) has been shown to be a simple, safe and cost-free delivery procedure that augments red cell mass in appropriate-for-gestational-age term and preterm

  9. A new clamp method for firing bricks | Obeng | Journal of Applied ...

    African Journals Online (AJOL)

    A conventional or standard clamp firing method has long been used by small and medium scale brick industries in Ghana, because the system is easier to build, cheaper to operate, can be constructed close to the supply of cl-ay and fuel to reduce transportation cost, and has a higher brick capacity of between 5.0 x 103 to ...

  10. Elastic and Plastic Analysis of a Single Edge Cracked Tension Specimen with Clamped Ends

    National Research Council Canada - National Science Library

    Mercier, Gerald

    1998-01-01

    ... (compliance and elastic stress intensity) and the plastic response (J-plastic, Eta(pl)) of the clamped end model were determined at ratios of height-to-width of 6, 8 and 10 and crack length to width ratios between 0.1 and 0.6...

  11. Proximal endovascular blood flow shunt for thoracoabdominal aortic aneurism without total aortic clamping

    Directory of Open Access Journals (Sweden)

    Gaudencio Espinosa

    Full Text Available The authors present a surgical approach to type III and IV Crawford aneurysms that does not need total aortic clamping, which allows the more objective prevention of direct ischemic damage, as well as its exclusion by the endoprosthesis implantation, shunting the flow to the synthetic graft.

  12. Gain-clamping techniques in two-stage double-pass L-band EDFA

    Indian Academy of Sciences (India)

    Two designs of long-wavelength band erbium-doped fiber amplifier (L-band EDFA) for gain clamping in double-pass systems are demonstrated and compared. The first design is based on ring laser technique where a backward amplified spontaneous emission (ASE) from the second stage is routed into the feedback loop ...

  13. space vector pulse width modulation of a multi-level diode clamped ...

    African Journals Online (AJOL)

    ES Obe

    nique in most industrial drives and control applications. It also focuses on the step by step ... the zero vector and two adjacent active vectors in determining the switching pro- cess of the multilevel DCC, quite unlike the .... Table 1: Switching states of an n-level Diode clamped converter. Switching States. S1. S2 ... Sn−2. Sa−1.

  14. Five-Level Active-Neutral-Point-Clamped DC/DC Converter

    DEFF Research Database (Denmark)

    Liu, Dong; Deng, Fujin; Chen, Zhe

    2016-01-01

    Multi-level converters are getting more and more attentions because of their obvious merits such as lower voltage stress and harmonic, smaller size of output filters, and so on. In this paper, a five-level active-neutral-point-clamped (5L-ANPC) dc/dc converter is proposed for power transfer...

  15. Seasonal growth of the attachment clamps of a Paradiplozoon sp. as ...

    African Journals Online (AJOL)

    Seasonal growth of the attachment clamps of a Paradiplozoon sp. as depicted by statistical shape analysis. SJ Milne, A Avenant-Oldewage. Abstract. Geometric morphometric methods using computer software is a more statistically powerful method of assessing changes in the anatomy than are traditional measurements of ...

  16. Two-Dimensional Fiber Positioning and Clamping Device for Product-Internal Microassembly

    NARCIS (Netherlands)

    Henneken, V.A.; Sassen, W.P.; Van der Vlist, W.; Wien, W.H.A.; Tichem, M.; Sarro, P.M.

    2008-01-01

    In this paper, we present a microelectromechanical systems-based two-degrees-of-freedom positioning device combined with a clamping structure for positioning and constraining an optical fiber. The fiber position can be controlled in the two directions perpendicular to the fiber axis using two

  17. space vector pulse width modulation of a multi-level diode clamped ...

    African Journals Online (AJOL)

    ES Obe

    Abstract. This paper presents the analysis of a multi-level diode clamped converter (DCC) which is implemented using space-vector pulse width modulation (SVPWM) tech- nique, widely considered as the most efficient and easily applied modulation tech- nique in most industrial drives and control applications.

  18. Glucose consumption by various tissues in pregnant rats : effects of a 6-day euglycaemic hyperinsulinaemic clamp

    NARCIS (Netherlands)

    Nieuwenhuizen, AG; Schuiling, GA; Bonen, A; Paans, AMJ; Vaalburg, W; Koiter, TR

    1998-01-01

    In the course of pregnancy maternal tissues become increasingly more insensitive to insulin. As 6 days of euglycaemic hyperinsulinaemic clamping, from day 8 until 14 of gestation, ameliorates total glucose consumption, we analysed the contribution of individual tissues in this phenomenon. We

  19. Diuretic response to acute hypertension is blunted during angiotensin II clamp

    DEFF Research Database (Denmark)

    Leong, Patrick K K; Zhang, Yibin; Yang, Li E

    2002-01-01

    whether the suppression of the renin-angiotensin system during acute hypertension affects the magnitude of the inhibition of PT fluid and sodium reabsorption, plasma ANG II levels were clamped by infusion of the angiotensin-converting enzyme (ACE) inhibitor captopril (12 microg/min) and ANG II after...

  20. MATLAB implementation of a dynamic clamp with bandwidth >125 KHz capable of generating INa at 37°C

    Science.gov (United States)

    Clausen, Chris; Valiunas, Virginijus; Brink, Peter R.; Cohen, Ira S.

    2012-01-01

    We describe the construction of a dynamic clamp with bandwidth >125 KHz that utilizes a high performance, yet low cost, standard home/office PC interfaced with a high-speed (16 bit) data acquisition module. High bandwidth is achieved by exploiting recently available software advances (code-generation technology, optimized real-time kernel). Dynamic-clamp programs are constructed using Simulink, a visual programming language. Blocks for computation of membrane currents are written in the high-level matlab language; no programming in C is required. The instrument can be used in single- or dual-cell configurations, with the capability to modify programs while experiments are in progress. We describe an algorithm for computing the fast transient Na+ current (INa) in real time, and test its accuracy and stability using rate constants appropriate for 37°C. We then construct a program capable of supplying three currents to a cell preparation: INa, the hyperpolarizing-activated inward pacemaker current (If), and an inward-rectifier K+ current (IK1). The program corrects for the IR drop due to electrode current flow, and also records all voltages and currents. We tested this program on dual patch-clamped HEK293 cells where the dynamic clamp controls a current-clamp amplifier and a voltage-clamp amplifier controls membrane potential, and current-clamped HEK293 cells where the dynamic clamp produces spontaneous pacing behavior exhibiting Na+ spikes in otherwise passive cells. PMID:23224681

  1. State-of-the-art automated patch clamp: heat activation, action potentials, and high throughput in ion channel screening.

    Science.gov (United States)

    Stoelzle-Feix, Sonja

    2014-01-01

    A successful robotic approach of the patch clamp technique is based on planar patch clamp chips where a glass pipette, as used in conventional patch clamping, is replaced by a thin planar glass sheet with a small hole in the middle. Automated patch clamp (APC) systems utilizing this chip design offer higher throughput capabilities and ease of use and thus have become common in basic research, drug development, and safety screening. Further development of existing devices and introduction of new systems widen the range of possible experiments and increase throughput. Here, two features with different areas of applications that meet the needs of drug discovery researchers and basic researchers alike are described. The utilized system is a medium throughput APC device capable of recording up to eight cells simultaneously. The temperature control capability and the possibility to perform recordings not only in the voltage clamp but also in the current clamp mode are described in detail. Since eight recordings can be generated in parallel without compromising data quality, reliable and cost-effective and time-effective screening of compounds against ion channels using voltage clamp and current clamp electrophysiology can be performed.

  2. Evaluation of diel patterns of relative changes in cell turgor of tomato plants using leaf patch clamp pressure probes

    NARCIS (Netherlands)

    Lee, K.M.; Driever, S.M.; Heuvelink, E.; Rüger, S.; Zimmermann, U.; Gelder, de A.; Marcelis, L.F.M.

    2012-01-01

    Relative changes in cell turgor of leaves of well-watered tomato plants were evaluated using the leaf patch clamp pressure probe (LPCP) under dynamic greenhouse climate conditions. Leaf patch clamp pressure changes, a measure for relative changes in cell turgor, were monitored at three different

  3. [Timing of clamping and factors associated with iron stores in full-term newborns].

    Science.gov (United States)

    Oliveira, Fabiana de Cássia Carvalho; Assis, Karine Franklin; Martins, Mariana Campos; Prado, Mara Rúbia Maciel Cardoso do; Ribeiro, Andréia Queiroz; Sant'Ana, Luciana Ferreira da Rocha; Priore, Silvia Eloiza; Franceschini, Sylvia do Carmo Castro

    2014-02-01

    To analyze the impact of timing of clamping and obstetric, biological and socioeconomic factors on the iron stores of full-term newborns. Cross-sectional study between October 2011 and July 2012 in which hematological parameters were evaluated for newborns in Viçosa, MG, Southeastern Brazil. It involved collecting 7 mL of umbilical cord blood from 144 full-term not underweight newborns. The parameters investigated were complete blood count, serum iron, ferritin and C-reactive protein. The time of umbilical cord clamping was measured using a digital timer without interfering in the procedures of childbirth. The birth data were collected from Live Birth Certificates and other information was obtained from the mother through a questionnaire applied in the first month postpartum. Analysis of multiple linear regression was then used to estimate the influence of biological, obstetrics and socioeconomic factors on the ferritin levels at birth. The median ferritin was 130.3 µg/L (n = 129, minimum = 16.4; maximum = 420.5 µg/L), the mean serum iron was 137.9 μg/dL (n = 144, SD = 39.29) and mean hemoglobin was 14.7 g/dL (n = 144, SD = 1.47). The median time of cord clamping was 36 seconds, ranging between 7 and 100. The bivariate analysis detected an association between ferritin levels and color of the child, timing clamping of 60 seconds, type of delivery, the presence of gestational diabetes and per capita family income. In multivariate analysis, the variables per capita income, number of antenatal visits and length at birth accounted for 22.0% of variation in ferritin levels. Iron stores at birth were influenced by biological, obstetric and social characteristics. Tackling anemia should involve creating policies aimed at reducing social inequalities, improving the quality of antenatal care, as well as implementing a criterion of delayed clamping of the umbilical cord within the guidelines of labor.

  4. Modified Levator Muscle Resection Using Putterman Muller's Muscle-Conjunctival Resection-Ptosis Clamp.

    Science.gov (United States)

    Yang, Ju-Wen

    2017-11-28

    The traditional technique for levator muscle resection includes a time-consuming dissection procedure that causes tissue trauma and swelling. The Putterman ptosis clamp has been popularly used in recent years for conjunctival müllerectomy. In this paper, we describe a modified surgical technique for ptosis treatment using the Putterman ptosis clamp. The modified technique improves the surgical results of levator muscle resection. We performed a retrospective case-series study to determine the outcomes and complications associated with the use of the Putterman ptosis clamp in levator muscle resection. Adults aged ≥18 years with moderate-to-severe ptosis underwent the modified technique for levator muscle resection. We first performed dissection to expose the aponeurosis and tarsus. Then, we placed the Putterman ptosis clamp to measure redundant aponeurotic and septal tissues and to perform the resection. Following the adjustment of the eyelid fissure, we refixed the levator muscle to the tarsus with 4-0 vicryl stitches. Seventeen patients (34 eyes) were included in the study. Of the 34 eyes, 31 (91.2%) experienced the complete resolution of ptosis after the surgery. One patient (2 eyes, 5.9%) had mild bilateral dermatochalasis and received revision surgery 6 months postoperative. One patient (1 eye, 2.9%) lost the crease of the left eye and received revision surgeries 2 and 6 months after the first surgery. No residual ptosis or severe adverse events were noted in the patients. Modified levator muscle resection using the Putterman ptosis clamp is an effective procedure for ptosis treatment. 4.

  5. Ventilation before Umbilical Cord Clamping improves the physiological transition at birth.

    Directory of Open Access Journals (Sweden)

    Sasmira eBhatt

    2014-10-01

    Full Text Available The transition from a fetus to a neonate at birth represents a critical phase in our life. Most infants make this transition without complications, but preterm infants usually require some form of assistance due to immature cardiopulmonary systems that predispose them to lifelong sequelae. As the incidence of preterm birth is increasing, there is now an urgent need for the development of management strategies that facilitate this transition, which will likely include improved strategies for the management of the maternal third stage of labour. For instance, recent studies on the physiological transition at birth have led to the discovery that establishing ventilation in the infant before the umbilical cord is clamped greatly stabilizes the cardiovascular transition at birth. While most benefits of delayed clamping have previously been attributed to an increase in placenta to infant blood transfusion, clearly there are other significant benefits for the infant, which are not well understood. Nevertheless, if ventilation can be established before cord clamping in a preterm infant, the large adverse changes in cardiac function that normally accompanies umbilical cord clamping can be avoided. As preterm infants have an immature cerebral vascular bed, large swings in cardiovascular function places them at high risk of cerebral vascular rupture and the associated increased risk of mortality and morbidity. In view of the impact that cord clamping has on the cardiovascular transition at birth, it is also time to re-examine some of the strategies used in the management of the third stage of labour. These include the appropriate timing of uterotonic administration in relation to delivery of the infant and placenta. As there is a lack of evidence on the effects these individual practices have on the infant, there is a necessity to improve our understanding of their impact in order to develop strategies that facilitate the transition to newborn life.

  6. Efficacy of tranexamic acid plus drain-clamping to reduce blood loss in total knee arthroplasty: A meta-analysis.

    Science.gov (United States)

    Zhang, Yan; Zhang, Jun-Wei; Wang, Bao-Hua

    2017-06-01

    Perioperative blood loss is still an unsolved problem in total knee arthroplasty (TKA). The efficacy of the preoperative use of tranexamic acid (TXA) plus drain-clamping to reduce blood loss in TKA has been debated. This meta-analysis aimed to illustrate the efficacy of TXA plus drain-clamping to reduce blood loss in patients who underwent a TKA. In February 2017, a systematic computer-based search was conducted in PubMed, EMBASE, Web of Science, the Cochrane Database of Systematic Reviews, and Google Scholar. Data from patients prepared for TKA in studies that compared TXA plus drain-clamping versus TXA alone, drain-clamping alone, or controls were retrieved. The primary endpoint was the need for transfusion. The secondary outcomes were total blood loss, blood loss in drainage, the decrease in hemoglobin, and the occurrence of deep venous thrombosis. After testing for publication bias and heterogeneity between studies, data were aggregated for random-effects models when necessary. Ultimately, 5 clinical studies with 618 patients (TXA plus drain-clamping group = 249, control group = 130, TXA-alone group = 60, and drain-clamping group = 179) were included. TXA plus drain-clamping could decrease the need for transfusion, total blood loss, blood loss in drainage, and the decrease in hemoglobin than could the control group, the TXA-alone group, and the drain-clamping group (P  .05). TXA plus drain-clamping can achieve the maximum effects of hemostasis in patients prepared for primary TKA. Because the number and the quality of the included studies were limited, more high-quality randomized controlled trials are needed to identify the optimal dose of TXA and the clamping hours in patients prepared for TKA.

  7. [How to measure insulin sensitivity in clinical practice?].

    Science.gov (United States)

    Rabasa-Lhoret, R; Laville, M

    2001-04-01

    Insulin resistance is common and cluster with glucose intolerance, dyslipidaemia and high blood pressure,. in type 2 diabetes mellitus it play a key role in the occurence of hyperglycemia. The importance of the insulin-resistant phenotype for the assessment of cardiovascular risk and response to intervention is increasingly being recognized. Therefore there is a need for accurate, reproducible and simple methods for measuring insulin resistance in vivo. The euglycemic hyperinsulinemic clamp is currently the best available standard technique but is not suitable for clinical practice. Thus, numerous index for insulin resistance estimation from fasting or post-load OGTT glycemia and insulinemia have been proposed. Although their simplicity is an obvious advantage, their application is subject to numerous limitations. The choice of the method to evaluate insulin sensitivity thus depend on objectives and available means. For clinical research, euglycemic clamp is the gold standard. In the case of epidemiologic studies, validated models like HOMA model are suitable. Finally in clinical practice, for type 2 diabetic patients, evaluation of insulin resistance should be made from clinical and biological context eventually associated with an estimation of respectives roles of insulinopenia and insulin resistance with a validated index like the HOMA model.

  8. Anti-Diabetic Activities of Gastrodia elata Blume Water Extracts Are Mediated Mainly by Potentiating Glucose-Stimulated Insulin Secretion and Increasing β-Cell Mass in Non-Obese Type 2 Diabetic Animals

    Directory of Open Access Journals (Sweden)

    Hye Jeong Yang

    2016-03-01

    Full Text Available The brain is an important modulator of glucose metabolism, and is known to respond Gastrodia elata Blume water extract (GEB. Therefore, we examined whether long-term administration of GEB has hypoglycemic activity, and its action mechanism was explored in partially-pancreatectomized rats that exhibit similar characteristics as Asian type 2 diabetes, non-obese insulin-insufficient diabetes. The rats were provided high-fat diets supplemented with either of (1 0.5% GEB (GEB-L, (2 2% GEB (GEB-H, (3 2% dextrin (control, or (4 2% dextrin with rosiglitazone (20 mg/kg body weight; positive-control for eight weeks. GEB dose-dependently improved hypothalamic insulin signaling, enhanced whole-body insulin sensitivity during hyperinsulinemic euglycemic clamp, and reduced hepatic glucose output in a hyperinsulinemic state. GEB dose-dependently increased the area under the curve of the serum insulin levels at the first and second phases during hyperglycemic clamp compared to the control, whereas the positive control had no effect. Insulin sensitivity during the hyperglycemic state also improved, dose-dependently, in response to GEB compared with that of the control, but was less than the positive control. GEB-H increased the mass of β-cells by potentiating proliferation and decreasing apoptosis. In conclusion, GEB could be a therapeutic agent for treating Asian type 2 diabetes.

  9. Capparis spinosa L. aqueous extract evokes antidiabetic effect in streptozotocin-induced diabetic mice

    Directory of Open Access Journals (Sweden)

    Mohamed Eddouks

    2017-02-01

    Full Text Available Objective: As the aqueous extract of Capparis spinosa (CS possess antidiabetic effect, he present study aims to reveal the possible  mechanism of action of CS in diabetic mice.Materials and Methods: Both single and repeated oral administrations of aqueous extract of CS were performed in multi-low dose streptozotocin-induced (MLDS diabetic mice. Euglycemic hyperinsulinemic clamp was used in association with the endogenous glucose production (perfusion rate of 3-3H glucose to evaluate the effect of CS aqueous extract on insulin sensitivity.Results: Our study showed that aqueous extract of CS possess a potent hypoglycaemic activity in MLDS diabetic mice. Furthermore, the analysis perfusion of 3-3H glucose demonstrated  the parallel decrease of basal endogenous glucose production (EGP with the hypoglycaemic activity. EGP was lower in CS-Treated group when compared to the control group (p

  10. Fitness, adiposopathy, and adiposity are independent predictors of insulin sensitivity in middle-aged men without diabetes.

    Science.gov (United States)

    Huth, Claire; Pigeon, Étienne; Riou, Marie-Ève; St-Onge, Josée; Arguin, Hélène; Couillard, Erick; Dubois, Marie-Julie; Marette, André; Tremblay, Angelo; Weisnagel, S John; Lacaille, Michel; Mauriège, Pascale; Joanisse, Denis R

    2016-09-01

    Adiposopathy, or sick fat, refers to adipose tissue dysfunction that can lead to several complications such as dyslipidemia, insulin resistance, and hyperglycemia. The relative contribution of adiposopathy in predicting insulin resistance remains unclear. We investigated the relationship between adiposopathy, as assessed as a low plasma adiponectin/leptin ratio, with anthropometry, body composition (hydrostatic weighing), insulin sensitivity (hyperinsulinemic-euglycemic clamp), inflammation, and fitness level (ergocycle VO2max, mL/kgFFM/min) in 53 men (aged 34-53 years) from four groups: sedentary controls without obesity (body mass index [BMI]  30 kg/m(2)), sedentary with obesity and glucose intolerance, and endurance trained active without obesity. The adiponectin/leptin ratio was the highest in trained men (4.75 ± 0.82) and the lowest in glucose intolerant subjects with obesity (0.27 ± 0.06; ANOVA p diabetes.

  11. Impact of Glucose Tolerance Status, Sex, and Body Size on Glucose Absorption Patterns During OGTTs

    DEFF Research Database (Denmark)

    Faerch, K.; Pacini, G.; Nolan, J. J.

    2013-01-01

    OBJECTIVEWe studied whether patterns of glucose absorption during oral glucose tolerance tests (OGTTs) were abnormal in individuals with impaired glucose regulation and whether they were related to sex and body size (height and fat-free mass). We also examined how well differences in insulin...... sensitivity and -cell function measured by gold-standard tests were reflected in the corresponding OGTT-derived estimates.RESEARCH DESIGN AND METHODSWith validated methods, various aspects of glucose absorption were estimated from 12-point, 3-h, 75-g OGTTs in 66 individuals with normal glucose tolerance (NGT......), isolated impaired fasting glucose (i-IFG), or isolated impaired glucose tolerance (i-IGT). Insulin sensitivity and -cell function were measured with the euglycemic-hyperinsulinemic clamp and intravenous glucose tolerance tests, respectively. Surrogate markers of both conditions were calculated from OGTTs...

  12. Regulation and function of FTO mRNA expression in human skeletal muscle and subcutaneous adipose tissue

    DEFF Research Database (Denmark)

    Grunnet, Louise G; Nilsson, Emma; Ling, Charlotte

    2009-01-01

    Objective. Common variants in FTO (the fat-mass and obesity-associated gene) associate with obesity and type 2 diabetes. The regulation and biological function of FTO mRNA expression in target tissue is unknown. We investigated the genetic and non-genetic regulation of FTO mRNA in skeletal muscle......) and elderly (58-66 years) non-diabetic twins examined by a hyperinsulinemic euglycemic clamp including indirect calorimetry. FTO mRNA expression was determined in subcutaneous adipose tissue (n=226) and skeletal muscle biopsies (n=158). Results. Heritability of FTO expression in both tissues was low, and FTO...... expression was not influenced by FTO rs9939609 genotype. FTO mRNA expression in skeletal muscle was regulated by age and sex, whereas age and BMI were predictors of adipose tissue FTO mRNA expression. FTO mRNA expression in adipose tissue was associated with an atherogenic lipid profile. In skeletal muscle...

  13. Impact of oral vancomycin on gut microbiota, bile acid metabolism, and insulin sensitivity

    DEFF Research Database (Denmark)

    Vrieze, Anne; Out, Carolien; Fuentes, Susana

    2014-01-01

    BACKGROUND & AIMS: Obesity has been associated with changes in the composition and function of the intestinal microbiota. Modulation of the microbiota by antibiotics also alters bile acid and glucose metabolism in mice. Hence, we hypothesized that short term administration of oral antibiotics...... in humans would affect fecal microbiota composition and subsequently bile acid and glucose metabolism. METHODS: In this single blinded randomized controlled trial, 20 male obese subjects with metabolic syndrome were randomized to 7 days of amoxicillin 500 mg t.i.d. or 7 days of vancomycin 500 mg t.......i.d. At baseline and after 1 week of therapy, fecal microbiota composition (Human Intestinal Tract Chip phylogenetic microarray), fecal and plasma bile acid concentrations as well as insulin sensitivity (hyperinsulinemic euglycemic clamp using [6,6-(2)H2]-glucose tracer) were measured. RESULTS: Vancomycin reduced...

  14. A high-fat, ketogenic diet causes hepatic insulin resistance in mice, despite increasing energy expenditure and preventing weight gain.

    Science.gov (United States)

    Jornayvaz, François R; Jurczak, Michael J; Lee, Hui-Young; Birkenfeld, Andreas L; Frederick, David W; Zhang, Dongyang; Zhang, Xian-Man; Samuel, Varman T; Shulman, Gerald I

    2010-11-01

    Low-carbohydrate, high-fat ketogenic diets (KD) have been suggested to be more effective in promoting weight loss than conventional caloric restriction, whereas their effect on hepatic glucose and lipid metabolism and the mechanisms by which they may promote weight loss remain controversial. The aim of this study was to explore the role of KD on liver and muscle insulin sensitivity, hepatic lipid metabolism, energy expenditure, and food intake. Using hyperinsulinemic-euglycemic clamps, we studied insulin action in mice fed a KD or regular chow (RC). Body composition was assessed by ¹H magnetic resonance spectroscopy. Despite being 15% lighter (P weight gain in mice, KD induces hepatic insulin resistance secondary to increased hepatic diacylglycerol content. Given the key role of nonalcoholic fatty liver disease in the development of type 2 diabetes and the widespread use of KD for the treatment of obesity, these results may have potentially important clinical implications.

  15. Genetic variation within the TRPM5 locus associates with prediabetic phenotypes in subjects at increased risk for type 2 diabetes

    DEFF Research Database (Denmark)

    Ketterer, Caroline; Müssig, Karsten; Heni, Martin

    2011-01-01

    genotyped 1798 white subjects at increased type 2 diabetes mellitus risk for 9 TRPM5 single nucleotide polymorphisms (namely, rs2301696, rs800344, rs800345, rs800347, rs800348, rs2074234, rs2301698, rs886277, and rs2301699) and also performed correlational analyses with metabolic traits. An oral glucose...... tolerance test (OGTT) was conducted on all subjects, and a subset (n = 525) additionally underwent a hyperinsulinemic-euglycemic clamp. The 9 chosen single nucleotide polymorphisms cover 100% of the common genetic variation (minor allele frequency =0.05) within the TRPM5 locus (D' = 1.0; r² = 0.8). Rs800344...... glucagon-like peptide-1 levels at 30 minutes during the OGTT compared with major allele homozygotes (P = .0124), whereas in male subjects, no significant differences were found (P = .3). In our German population, the common TRPM5 variants are likely to be associated with prediabetic phenotypes...

  16. Characterisation of adiponectin multimers and the IGF axis in humans with a heterozygote mutation in the tyrosine kinase domain of the insulin receptor gene

    DEFF Research Database (Denmark)

    Højlund, Kurt; Beck-Nielsen, Henning; Flyvbjerg, Allan

    2012-01-01

    Objective: Low levels of adiponectin, IGF-binding protein (IGFBP)-1, and IGFBP-2, and high levels of leptin correlate with several indices of insulin resistance and risk of type 2 diabetes. However, in insulin receptoropathies plasma adiponectin is paradoxically increased despite severe insulin...... resistance, whereas the IGF-axis is sparsely described. Here, we aimed to characterize the multimeric distribution of adiponectin and the IGF-axis in humans with a heterozygous INSR mutation (Arg1174Gln).Methods: Blood samples obtained in six Arg1174Gln-carriers and 10 lean, healthy controls before and after...... an euglycemic-hyperinsulinemic clamp were examined for plasma adiponectin multimers, leptin, total IGF-I, IGF-II, free IGF-I, IGFBP-1 and IGFBP-2.Results: Despite 10-fold elevated fasting insulin and marked insulin resistance in Arg1174Gln-carriers, the levels of total adiponectin, leptin, IGFBP-1 and IGFBP-2...

  17. Catecholamine and insulin control of lipolysis in subcutaneous adipose tissue during long-term diet-induced weight loss in obese women

    DEFF Research Database (Denmark)

    Koppo, Katrien; Siklová-Vitková, Michaela; Klimcáková, Eva

    2012-01-01

    low-calorie diet (LCD) and 3-mo weight maintenance (WM) diet. At each phase of the dietary intervention, microdialysis of subcutaneous adipose tissue (SCAT) was performed at rest and during a 3-h hyperinsulinemic euglycemic clamp. Responses of dialysate glycerol concentration (DGC) were determined...... the different phases of the diet. The adrenaline-induced increase in DGC was higher at VLCD and LCD compared with baseline condition and returned to prediet levels at WM. In the probe with adrenaline and phentolamine, the increase in DGC was higher than that in the adrenaline probe at baseline and WM......, but it was not different at VLCD and LCD. The results suggest that the responsiveness of SCAT to adrenaline-stimulated lipolysis increases during the calorie-restricted phases due to a reduction of the α(2)-adrenoceptor-mediated antilipolytic action of adrenaline. At WM, adrenaline-stimulated lipolysis returned...

  18. The reduction in visceral fat mass in response to growth hormone is more marked in men than in oestrogen-deficient women

    DEFF Research Database (Denmark)

    Franco, Celina; Koranyi, Josef; Brandberg, John

    2008-01-01

    CONTEXT: Women with severe growth hormone (GH) deficiency have a less marked response to GH replacement than men. This has mostly been attributed to the attenuating effects of oestrogen replacement therapy. OBJECTIVE: To study gender related differences in the response to GH treatment in men...... and postmenopausal women. METHODS: Fifteen men and 15 age- and BMI-matched women with abdominal obesity (mean age: 58; range 51-64 years) were treated for one year with similar doses (0.47 vs. 0.51 mg/day) of GH. All women were postmenopausal not receiving oestrogen treatment. Insulin sensitivity was assessed using...... a hyperinsulinemic euglycemic clamp and body composition by computed tomography (CT) scans and from total body potassium, K(40). RESULTS: Men and women were comparable at baseline in terms of waist circumference, IGF-1 and lipid levels. After one year of GH treatment, there was a 18% reduction in visceral adipose...

  19. The Dual Amylin- and Calcitonin-Receptor Agonist KBP-042 Increases Insulin Sensitivity and Induces Weight Loss in Rats with Obesity

    DEFF Research Database (Denmark)

    Hjuler, Sara Toftegaard; Gydesen, Sofie; Andreassen, Kim Vietz

    2016-01-01

    Objective: In this study, KBP-042, a dual amylin- and calcitonin-receptor agonist, was investigated as a treatment of obesity and insulin resistance in five different doses (0.625 μg/kg-10 μg/kg) compared with saline-treated and pair-fed controls. Methods: Rats with obesity received daily s...... combines two highly relevant features, namely weight loss and insulin sensitivity, and is thus an excellent candidate for chronic treatment of obesity and insulin resistance........c. administrations for 56 days, and glucose tolerance was assessed after one acute injection, 3 weeks of treatment, and again after 7 weeks of treatment. To assess the effect on insulin sensitivity, rats received 5 μg/kg KBP-042 for 21 days before hyperinsulinemic-euglycemic clamp. Results: KBP-042 induced...

  20. Insulin action in human thighs after one-legged immobilization

    DEFF Research Database (Denmark)

    Richter, Erik; Kiens, Bente; Mizuno, M.

    1989-01-01

    Insulin action was assessed in thighs of five healthy young males who had one knee immobilized for 7 days by a splint. The splint was not worn in bed. Subjects also used crutches to prevent weight bearing of the immobilized leg. Immobilization decreased the activity of citrate synthase and 3-OH......-acyl-CoA-dehydrogenase in the vastus lateralis muscle by 9 and 14%, respectively, and thigh volume by 5%. After 7 days of immobilization, a two-step euglycemic hyperinsulinemic clamp procedure combined with arterial and bilateral femoral venous catheterization was performed. Insulin action on glucose uptake and tyrosine release...... was significantly higher in the immobilized than in the control thigh. Seven days of one-legged immobilization causes local decreased insulin action on thigh glucose uptake and net protein degradation....

  1. Changes in insulin sensitivity precede changes in body composition during 14 days of step reduction combined with overfeeding in healthy young men

    DEFF Research Database (Denmark)

    Knudsen, Sine Haugaard; Hansen, Louise Seier; Pedersen, Maria

    2012-01-01

    A lifestyle characterized by inactivity and a high-calorie diet is a known risk factor for impaired insulin sensitivity and development of Type 2 diabetes mellitus. To investigate possible links, nine young healthy men (24 ± 3 yr; body mass index of 21.6 ± 2.5 kg/m(2)) completed 14 days of step...... reduction (10,000 to 1,500 steps/day) and overfeeding (+50% kcal). Body composition (dual X-ray absorptiometry, MRI), aerobic fitness (maximal O(2) consumption), systemic inflammation and insulin sensitivity [oral glucose tolerance test (OGTT), hyperinsulinemic euglycemic clamp] were assessed before (day 0......), during (days 3 and 7), and immediately after the intervention (day 14), with follow-up tests (day 30). Body weight had increased at days 7 and 14 (P day 14 compared with day 0 (P days 7 and 14 (P...

  2. Differential impact of selective GH deficiency and endogenous GH excess on insulin-mediated actions in muscle and liver of male mice.

    Science.gov (United States)

    Cordoba-Chacon, Jose; Gahete, Manuel D; McGuinness, Owen P; Kineman, Rhonda D

    2014-11-15

    A reciprocal relationship between insulin sensitivity and glucose tolerance has been reported in some mouse models and humans with isolated changes in growth hormone (GH) production and signaling. To determine if this could be explained in part by tissue-specific changes in insulin sensitivity, hyperinsulinemic-euglycemic clamps were performed in mice with adult-onset, isolated GH deficiency and in mice with elevated endogenous GH levels due to somatotrope-specific loss of IGF-I and insulin receptors. Our results demonstrate that circulating GH levels are negatively correlated with insulin-mediated glucose uptake in muscle but positively correlated with insulin-mediated suppression of hepatic glucose production. A positive relationship was also observed between GH levels and endpoints of hepatic lipid metabolism known to be regulated by insulin. These results suggest hepatic insulin resistance could represent an early metabolic defect in GH deficiency.

  3. Pioglitazone enhances expression of genes involved in mitochondrial oxidative metabolism in skeletal muscle of women with polycystic ovary syndrome (PCOS)

    DEFF Research Database (Denmark)

    Skov, Vibe

    Aims                Polycystic ovary syndrome (PCOS) is a common endocrine disorder in premenopausal women and is associated with insulin resistance increasing the risk for developing type 2 diabetes mellitus. Studies have shown that thiazolidinediones (TZD) improve metabolic disturbances in PCOS...... patients. We hypothesized that the effect of TZD in PCOS is in part mediated by changes in the transcriptional profile of muscle favoring insulin sensitivity. Methods Using the HG-U133 2.0 Plus expression array from Affymetrix, we examined the effect of pioglitazone (30 mg/day for 16 weeks) on gene...... expression in skeletal muscle of 10 obese women with PCOS (dataset 1). Furthermore, evaluation of gene expression changes between PCOS patients before treatment and control subjects were performed (dataset 2). All subjects were metabolically characterised by a euglycemic-hyperinsulinemic clamp combined...

  4. Skeletal Muscle Mitochondrial Function in Polycystic Ovarian Syndrome

    DEFF Research Database (Denmark)

    Rabøl, Rasmus; Svendsen, Pernille Maj; Skovbro, Mette

    2011-01-01

    Objective Polycystic ovarian syndrome (PCOS) is associated with skeletal muscle insulin resistance, which has been linked to decreased mitochondrial function. We measured mitochondrial respiration in lean and obese women with and without PCOS using high-resolution respirometry. Methods...... Hyperinsulinemic euglycemic clamps (40 mU/min/m2) and muscle biopsies were performed on 23 women with PCOS (9 lean (body mass index (BMI) 25 kg/m2)) and 17 age- and weight-matched controls (6 lean and 11 obese). Western blotting and high-resolution respirometry was used to determine mitochondrial function. Results...... Insulin sensitivity decreased with PCOS and increasing body weight. Mitochondrial respiration with substrates for complex I and complex I+II were similar in all groups, and PCOS was not associated with a decrease in mitochondrial content as measured by mtDNA/genomicDNA. We found no correlation between...

  5. Differential Nongenetic Impact of Birth Weight Versus Third-Trimester Growth Velocity on Glucose Metabolism and Magnetic Resonance Imaging Abdominal Obesity in Young Healthy Twins

    DEFF Research Database (Denmark)

    Pilgaard, Kasper; Mosbech, Thomas Hammershaimb; Grunnet, Louise

    2011-01-01

    monozygotic twins are instrumental in determining nongenetic associations between early environment and adult metabolic phenotype.Objective: Our objective was to investigate the relationship between size at birth and third-trimester growth velocity on adult body composition and glucose metabolism using intra......-pair differences in young healthy twins.Methods: Fifty-eight healthy twins (42 monozygotic/16 dizygotic) aged 18-24 yr participated. Insulin sensitivity was assessed using hyperinsulinemic-euglycemic clamps. Whole-body fat was assessed by dual-energy x-ray absorptiometry scan, whereas abdominal visceral and sc fat...... (L1-L4) were assessed by magnetic resonance imaging. Third-trimester growth velocity was determined by repeated ultrasound examinations.Results: Size at birth was nongenetically inversely associated with adult visceral and sc fat accumulation but unrelated to adult insulin action. In contrast, fetal...

  6. The effect of menopause on carotid artery remodeling, insulin sensitivity, and plasma adiponectin in healthy women

    DEFF Research Database (Denmark)

    Muscelli, Elza; Kozàkovà, Michaela; Flyvbjerg, Allan

    2009-01-01

    secretion and sensitivity, plasma adiponectin), and carotid intima-media thickness (IMT) in healthy women. METHODS: In 74 menopausal women (mean age = 51 +/- 3 years, mean duration of menopause = 2.9 +/- 1.2 years) and in 74 nonmenopausal women comparable for age and body mass index (BMI), common carotid...... artery (CCA) luminal diameter, and IMT in different carotid segments were measured in digitized ultrasound images. Insulin sensitivity and secretion were assessed using the euglycemic hyperinsulinemic clamp technique and oral glucose tolerance test (OGTT). Insulin secretion was reconstructed...... by mathematical modeling. RESULTS: CCA diameter (5.55 +/- 0.46 vs. 5.21+/- 0.51 mm, P IMT (608 +/- 78 vs. 576 +/- 74 microm, P IMT/diameter ratio and IMT in other carotid...

  7. Fatty liver is associated with insulin resistance, risk of coronary heart disease, and early atherosclerosis in a large European population

    DEFF Research Database (Denmark)

    Gastaldelli, Amalia; Kozakova, Michaela; Højlund, Kurt

    2009-01-01

    ,307 nondiabetic subjects (age 30-60 years) recruited at 19 centers, we evaluated liver enzymes, lipids, insulin sensitivity (by euglycemic-hyperinsulinemic clamp), glucose tolerance (by 75 g oral glucose tolerance test), carotid atherosclerosis as intima media thickness (IMT), CHD risk by the Framingham Heart......: intermediate group (n = 465). Compared to G1, G3 included more men (70% versus 24%) and people with impaired glucose tolerance (23% versus 5%). IMT increased with FLI (G3 = 0.64 +/- 0.08 versus G1 = 0.58 +/- 0.08 mm, P ... cholesterol (r = 0.33), alanine aminotransferase (r = 0.48), aspartate aminotransferase (r = 0.25), systolic blood pressure (r = 0.39) and IMT (r = 0.30), and reduced insulin sensitivity (r = -0.43), high-density lipoprotein cholesterol (r = -0.50), adiponectin (r = -0.42), and physical activity (r = -0...

  8. Testosterone therapy increased muscle mass and lipid oxidation in aging men

    DEFF Research Database (Denmark)

    Frederiksen, Louise; Højlund, Kurt; Hougaard, David M

    2011-01-01

    The indication for testosterone therapy in aging hypogonadal men without hypothalamic, pituitary, or testicular disease remains to be elucidated. The aim of this study was to investigate the effect of testosterone therapy on insulin sensitivity, substrate metabolism, body composition, and lipids...... in aging men with low normal bioavailable testosterone levels using a predefined cutoff level for bioavailable testosterone. A randomized, double-blinded, placebo-controlled study of testosterone treatment (gel) was done on 38 men, aged 60-78 years, with bioavailable testosterone 94 cm. Insulin......-stimulated glucose disposal (Rd) and substrate oxidation were assessed by euglycemic hyperinsulinemic clamps combined with indirect calorimetry. Lean body mass (LBM) and total fat mass (TFM) were measured by dual x-ray absorptiometry, and serum total testosterone was measured by tandem mass spectrometry...

  9. Expression of insulin receptor spliced variants and their functional correlates in muscle from patients with non-insulin-dependent diabetes mellitus

    DEFF Research Database (Denmark)

    Hansen, Torben; Bjørbaek, C; Vestergaard, H

    1993-01-01

    Due to alternative splicing of exon 11 of the receptor gene, the human insulin receptor exists in two forms, that have distinct tissue-specific expression and are functionally different. Needle biopsies obtained from vastus lateralis muscle from 20 patients with noninsulin-dependent diabetes...... mellitus (NIDDM) and 20 normal control subjects were analyzed for the relative expression of insulin receptor mRNA variants in a novel assay using fluorescence-labeled primers and subsequent analysis on an automated DNA sequencer. In subgroups of patients and control subjects, insulin binding and tyrosine...... kinase activity were examined in wheat germ agglutinin-purified insulin receptors isolated from muscle biopsies. Moreover, insulin-stimulated glucose disposal was studied by means of the euglycemic hyperinsulinemic clamp technique. No difference in the relative expression of spliced variants...

  10. Dysregulation of glycogen synthase COOH- and NH2-terminal phosphorylation by insulin in obesity and type 2 diabetes mellitus

    DEFF Research Database (Denmark)

    Højlund, Kurt; Birk, Jesper Bratz; Klein, Ditte Kjærsgaard

    2009-01-01

    , phospholamban-Thr17, was also evident in T2DM. Dephosphorylation of GS by phosphatase treatment fully restored GS activity in all groups. Conclusions: Dysregulation of GS phosphorylation plays a major role in impaired insulin regulation of GS in obesity and T2DM. In obesity, independent of T2DM......Context: Insulin-stimulated glucose disposal is impaired in obesity and type 2 diabetes mellitus (T2DM) and is tightly linked to impaired skeletal muscle glucose uptake and storage. Impaired activation of glycogen synthase (GS) by insulin is a well-established defect in both obesity and T2DM......, but the underlying mechanisms remain unclear. Design and Participants: Insulin action was investigated in a matched cohort of lean healthy, obese nondiabetic, and obese type 2 diabetic subjects by the euglycemic-hyperinsulinemic clamp technique combined with muscle biopsies. Activity, site-specific phosphorylation...

  11. Pioglitazone enhances mitochondrial biogenesis and ribosomal protein biosynthesis in skeletal muscle in polycystic ovary syndrome

    DEFF Research Database (Denmark)

    Skov, Vibe; Glintborg, Dorte; Knudsen, Steen

    2008-01-01

    Insulin resistance is a common metabolic abnormality in women with PCOS and leads to an elevated risk of type 2 diabetes. Studies have shown that thiazolidinediones (TZDs) improve metabolic disturbances in PCOS patients. We hypothesized that the effect of TZDs in PCOS is, in part, mediated...... by changes in the transcriptional profile of muscle favoring insulin sensitivity. Using Affymetrix microarrays, we examined the effect of pioglitazone (30 mg/day for 16 weeks) on gene expression in skeletal muscle of 10 obese women with PCOS metabolically characterized by a euglycemic-hyperinsulinemic clamp....... Moreover, we explored gene expression changes between these PCOS patients before treatment and 13 healthy women. Treatment with pioglitazone improved insulin-stimulated glucose metabolism and plasma adiponectin, and reduced fasting serum insulin (all P

  12. IRS-1 serine phosphorylation and insulin resistance in skeletal muscle from pancreas tranplant recipient

    DEFF Research Database (Denmark)

    Bouzakri, K; Karlsson, HRK; Vestergaard, Henrik

    2006-01-01

    Insulin-dependent diabetic recipients of successful pancreas allografts achieve self-regulatory insulin secretion and discontinue exogenous insulin therapy; however, chronic hyperinsulinemia and impaired insulin sensitivity generally develop. To determine whether insulin resistance is accompanied...... by altered signal transduction, skeletal muscle biopsies were obtained from pancreas-kidney transplant recipients (n = 4), nondiabetic kidney transplant recipients (receiving the same immunosuppressive drugs; n = 5), and healthy subjects (n = 6) before and during a euglycemic-hyperinsulinemic clamp. Basal...... insulin receptor substrate (IRS)-1 Ser (312) and Ser (616) phosphorylation, IRS-1-associated phosphatidylinositol 3-kinase activity, and extracellular signal-regulated kinase (ERK)-1/2 phosphorylation were elevated in pancreas-kidney transplant recipients, coincident with fasting hyperinsulinemia. Basal...

  13. Hypoglycemia-Related Electroencephalogram Changes Assessed by Multiscale Entropy

    DEFF Research Database (Denmark)

    Fabris, C.; Sparacino, G.; Sejling, A. S.

    2014-01-01

    Background: Several clinical studies have shown that low blood glucose (BG) levels affect electroencephalogram (EEG) rhythms through the quantification of traditional indicators based on linear spectral analysis. Nonlinear measures used in the last decades to characterize the EEG in several...... physiopathological conditions have never been assessed in hypoglycemia. The present study investigates if properties of the EEG signal measured by nonlinear entropy-based algorithms are altered in a significant manner when a state of hypoglycemia is entered. Subjects and Methods: EEG was acquired from 19 patients...... with type 1 diabetes during a hyperinsulinemic-euglycemic-hypoglycemic clamp experiment. In parallel, BG was frequently monitored by the standard YSI glucose and lactate analyzer and used to identify two 1-h intervals corresponding to euglycemia and hypoglycemia, respectively. In each subject, the P3-C3 EEG...

  14. The adipose transcriptional response to insulin is determined by obesity, not insulin sensitivity

    DEFF Research Database (Denmark)

    Rydén, Mikael; Hrydziuszko, Olga; Mileti, Enrichetta

    2016-01-01

    Metabolically healthy obese subjects display preserved insulin sensitivity and a beneficial white adipose tissue gene expression pattern. However, this observation stems from fasting studies when insulin levels are low. We investigated adipose gene expression by 5'Cap-mRNA sequencing in 17 healthy...... non-obese (NO), 21 insulin-sensitive severely obese (ISO), and 30 insulin-resistant severely obese (IRO) subjects, before and 2 hr into a hyperinsulinemic euglycemic clamp. ISO and IRO subjects displayed a clear but globally similar transcriptional response to insulin, which differed from the small...... effects observed in NO subjects. In the obese, 231 genes were altered; 71 were enriched in ISO subjects (e.g., phosphorylation processes), and 52 were enriched in IRO subjects (e.g., cellular stimuli). Common cardio-metabolic risk factors and gender do not influence these findings. This study demonstrates...

  15. Computational model of cellular metabolic dynamics

    DEFF Research Database (Denmark)

    Li, Yanjun; Solomon, Thomas; Haus, Jacob M

    2010-01-01

    , by application of mechanism M.3, the model predicts metabolite concentration changes and glucose partitioning patterns consistent with experimental data. The reaction rate fluxes quantified by this detailed model of insulin/glucose metabolism provide information that can be used to evaluate the development......, pyruvate dehydrogenase); or M.3, parallel activation by a phenomenological insulin-mediated intracellular signal that modifies reaction rate coefficients. These simulations indicated that models M.1 and M.2 were not sufficient to explain the experimentally measured metabolic responses. However...... of the cytosol and mitochondria. The model simulated skeletal muscle metabolic responses to insulin corresponding to human hyperinsulinemic-euglycemic clamp studies. Insulin-mediated rate of glucose disposal was the primary model input. For model validation, simulations were compared with experimental data...

  16. Reduced CD300LG mRNA tissue expression, increased intramyocellular lipid content and impaired glucose metabolism in healthy male carriers of Arg82Cys in CD300LG a novel genometabolic cross-link between CD300LG and common metabolic phenotypes

    DEFF Research Database (Denmark)

    Støy, Julie; Kampmann, Ulla; Mengel, Annette

    2015-01-01

    by the hyperinsulinemic euglycemic clamp, intrahepatic lipid content (IHLC) and intramyocellular lipid content (IMCL) by MR spectroscopy, and β-cell function by an intravenous glucose tolerance test. Changes in insulin signaling and CD300LG mRNA expression were determined by western blotting and quantitative PCR...... with extracellular lipids. We tested whether this specific polymorphism results in abnormal lipid accumulation in skeletal muscle and liver and other indices of metabolic dysfunction. METHODS: 40 healthy men with a mean age of 55 years were characterized metabolically including assessment of insulin sensitivity......: The CD300LG polymorphism was associated with decreased CD300LG mRNA expression in muscle and adipose tissue, increased IMCL, and abnormalities of glucose metabolism. CD300LG mRNA levels correlated with IMCL and forearm glucose uptake. These findings link a specific CD300LG polymorphism with features...

  17. Insulin resistance and maximal oxygen uptake

    DEFF Research Database (Denmark)

    Seibaek, Marie; Vestergaard, Henrik; Burchardt, Hans

    2003-01-01

    BACKGROUND: Type 2 diabetes, coronary atherosclerosis, and physical fitness all correlate with insulin resistance, but the relative importance of each component is unknown. HYPOTHESIS: This study was undertaken to determine the relationship between insulin resistance, maximal oxygen uptake......, and the presence of either diabetes or ischemic heart disease. METHODS: The study population comprised 33 patients with and without diabetes and ischemic heart disease. Insulin resistance was measured by a hyperinsulinemic euglycemic clamp; maximal oxygen uptake was measured during a bicycle exercise test. RESULTS......: There was a strong correlation between maximal oxygen uptake and insulin-stimulated glucose uptake (r = 0.7, p = 0.001), and maximal oxygen uptake was the only factor of importance for determining insulin sensitivity in a model, which also included the presence of diabetes and ischemic heart disease. CONCLUSION...

  18. Pre- and posttranslational upregulation of muscle-specific glycogen synthase in athletes

    DEFF Research Database (Denmark)

    Vestergaard, H; Andersen, P H; Lund, S

    1994-01-01

    Expression of muscle-specific glycogen synthase (GS) and phosphofructokinase (PFK) was analyzed in seven athletes and eight control subjects who were characterized using the euglycemic, hyperinsulinemic (2 mU.kg-1.min-1) clamp technique in combination with indirect calorimetry and biopsy sampling.......005) and both nonoxidative (P metabolism were significantly higher in athletes. In parallel, after hyperinsulinemia, the relative activation of GS by G-6-P was significantly higher in athletes, whereas total activity and gene expression of both GS and PFK were unaffected...... by insulin. We conclude that athletes have increased whole body insulin-stimulated nonoxidative glucose metabolism associated with both pretranslational (mRNA) and posttranslational (enzyme activity) upregulation of GS. However, the immunoreactive mass of GS is normal, emphasizing that posttranslational...

  19. Effects of exercise training and diet on lipid kinetics during free fatty acid-induced insulin resistance in older obese humans with impaired glucose tolerance

    DEFF Research Database (Denmark)

    Solomon, Thomas; Haus, Jacob M; Marchetti, Christine M

    2009-01-01

    Elevated free fatty acids (FFA) are implicated with insulin resistance at the cellular level. However, the contribution of whole body lipid kinetics to FFA-induced insulin resistance is not well understood, and the effect of exercise and diet on this metabolic defect is not known. We investigated.......8 +/- 1.8 kg/m(2)) or a eucaloric (n = 8; 67 +/- 2 yr, 35.3 +/- 2.1 kg/m(2)) diet and aerobic exercise (1 h/day at 65% of maximal oxygen uptake) regimen. Lipid kinetics ([1-(14)C]palmitate) were assessed throughout a 7-h, 40 mU x m(-2) x min(-1) hyperinsulinemic euglycemic clamp, during which insulin...

  20. Autonomic nervous system activation mediates the increase in whole-body glucose uptake in response to electroacupuncture

    DEFF Research Database (Denmark)

    Benrick, Anna; Kokosar, Milana; Hu, Min

    2017-01-01

    measured by euglycemic-hyperinsulinemic clamp during and after 45 min of low-frequency EA in 21 overweight/obese women with polycystic ovary syndrome (PCOS) and 21 controls matched for age, weight, and body mass index (experiment 1) and in rats receiving autonomic receptor blockers (experiment 2). GIR...... was higher after EA in controls and women with PCOS. Plasma serotonin levels and homovanillic acid, markers of vagal activity, decreased in both controls and patients with PCOS. Adipose tissue expression of pro-nerve growth factor (proNGF) decreased, and the mature NGF/proNGF ratio increased after EA in PCOS...... of EA increases whole-body glucose uptake by activation of the sympathetic and partly the parasympathetic nervous systems, which could have important clinical implications for the treatment of insulin resistance.-Benrick, A., Kokosar, M., Hu, M., Larsson, M., Maliqueo, M., Marcondes, R. R., Soligo, M...

  1. Blueberries’ Impact on Insulin Resistance and Glucose Intolerance

    Directory of Open Access Journals (Sweden)

    April J. Stull

    2016-11-01

    Full Text Available Blueberries are a rich source of polyphenols, which include anthocyanin bioactive compounds. Epidemiological evidence indicates that incorporating blueberries into the diet may lower the risk of developing type 2 diabetes (T2DM. These findings are supported by pre-clinical and clinical studies that have shown improvements in insulin resistance (i.e., increased insulin sensitivity after obese and insulin-resistant rodents or humans consumed blueberries. Insulin resistance was assessed by homeostatic model assessment-estimated insulin resistance (HOMA-IR, insulin tolerance tests, and hyperinsulinemic-euglycemic clamps. Additionally, the improvements in glucose tolerance after blueberry consumption were assessed by glucose tolerance tests. However, firm conclusions regarding the anti-diabetic effect of blueberries cannot be drawn due to the small number of existing clinical studies. Although the current evidence is promising, more long-term, randomized, and placebo-controlled trials are needed to establish the role of blueberries in preventing or delaying T2DM.

  2. Increased recovery rates of phosphocreatine and inorganic phosphate after isometric contraction in oxidative muscle fibres and elevated hepatic insulin resistance in homozygous carriers of the A-allele of FTO rs9939609

    DEFF Research Database (Denmark)

    Grunnet, Louise Groth; Brøns, Charlotte; Jacobsen, Stine

    2009-01-01

    diabetes. Methods. Forty-six young men underwent a hyperinsulinemic euglycemic clamp with excision of skeletal muscle biopsies, an intravenous glucose tolerance test, (31)phosphorous magnetic resonance spectroscopy and 24-hour whole body metabolism was measured in a respiratory chamber. Results. The FTO rs......9939609 A-allele was associated with elevated fasting blood glucose and plasma insulin, hepatic insulin resistance and shorter recovery halftimes of phosphocreatine (PCr) and inorganic phosphate (Pi) after exercise in a primarily type I muscle. These relationships - except for fasting insulin - remained...... in homozygous carriers of the FTO A-risk-allele. Hepatic insulin resistance may represent the key metabolic defect responsible for mild elevations of fasting blood glucose associated to the FTO phenotype....

  3. Acute hyperinsulinemia is followed by increased serum concentrations of fibroblast growth factor 23 in type 2 diabetes patients

    DEFF Research Database (Denmark)

    Winther, Kristian; Nybo, Mads; Vind, Birgitte

    2012-01-01

    Background. Phosphate homeostasis is connected to glucose metabolism and is influenced by insulin, but the role of fibroblast growth factor 23 (FGF23) is unknown in this relation. Therefore, the levels of FGF23 and phosphate were investigated during a euglycemic-hyperinsulinemic clamp in healthy...... for the understanding of phosphate metabolism in relation to type 2 diabetes and its vascular complications.......)) to determine glucose disposal rates. Blood samples for measurement of serum FGF23 and insulin, plasma phosphate and glucose, and HbA(1c) were drawn before and after insulin infusion. Results. The three groups did not differ in baseline levels of serum FGF23. Insulin infusion increased serum FGF23...

  4. Severe insulin-resistant diabetes mellitus in patients with congenital muscle fiber type disproportion myopathy

    DEFF Research Database (Denmark)

    Vestergaard, H; Klein, H H; Hansen, T

    1995-01-01

    severe insulin resistance of both liver and peripheral tissues. The impaired insulin-stimulated glucose disposal to peripheral tissues was primarily due to reduced nonoxidative glucose metabolism. These changes were paralleled by reduced basal values of muscle GS total activity, allosterical activation...... heterogeneity. Concomitant endocrine dysfunctions have not been described. We report the findings of altered insulin secretion and insulin action in two brothers affected with CFTDM and glucose intolerance as well as in their nonconsanguineous glucose-tolerant parents. Results are compared with those of six...... normoglycemic control subjects. All study participants underwent an oral glucose tolerance test to estimate insulin secretion. The oldest boy and his parents volunteered for studies of whole-body insulin sensitivity consisting of a 4-h euglycemic hyperinsulinemic clamp in combination with indirect calorimetry...

  5. Differential effects of glucagon-like peptide-1 on microvascular recruitment and glucose metabolism in short- and long-term Insulin resistance

    DEFF Research Database (Denmark)

    Sjøberg, Kim Anker; Rattigan, Stephen; Jeppesen, Jacob Fuglsbjerg

    2015-01-01

    Acute infusion of glucagon-like-peptide-1 (GLP-1) has potent effects on blood flow distribution through the microcirculation in healthy humans and rats. High fat diet induces impairments in insulin-mediated microvascular recruitment (MVR) and muscle glucose uptake, and here we examined whether...... this could be reversed by GLP-1. Using contrast-enhanced ultrasound, microvascular recruitment was assessed by continuous real-time imaging of gas-filled microbubbles in the microcirculation after acute (5 days) and prolonged (8 weeks) high fat diet (HF) induced insulin resistance in rats. An euglycemic...... hyperinsulinemic clamp (3 mU·min(-1) ·kg(-1) ) with or without a co-infusion of GLP-1 (100 pmol·l(-1) ) was performed in anaesthetized rats. Consumption of the HF diet attenuated the insulin-mediated MVR in both 5 days and 8 weeks HF interventions which was associated with a 50% reduction in insulin...

  6. Gender differences in skeletal muscle substrate metabolism - molecular mechanisms and insulin sensitivity

    DEFF Research Database (Denmark)

    Lundsgaard, Annemarie; Kiens, Bente

    2014-01-01

    higher insulin sensitivity of female skeletal muscle can be related to gender-specific regulation of molecular metabolism will be topic for discussion. Gender differences in muscle fiber type distribution and substrate availability to and in skeletal muscle are highly relevant for substrate metabolism......It has become increasingly apparent that substrate metabolism is subject to gender-specific regulation, and the aim of this review is to outline the available evidence of molecular gender differences in glucose and lipid metabolism of skeletal muscle. Female sex has been suggested to have...... a favorable effect on glucose homeostasis, and the available evidence from hyperinsulinemic-euglycemic clamp studies is summarized to delineate whether there is a gender difference in whole-body insulin sensitivity and in particular insulin-stimulated glucose uptake of skeletal muscle. Whether an eventual...

  7. Genetically Determined Insulin Resistance is Characterized by Down-Regulation of Mitochondrial Oxidative Metabolism in Human Skeletal Muscle

    DEFF Research Database (Denmark)

    Kristensen, Jonas M; Skov, Vibe; Wojtaszewski, Jørgen

    2010-01-01

    mitochondrial dysfunction is a cause or consequence of insulin resistance remains to be clarified. In the present study, we tested the hypothesis that mitochondrial oxidative metabolism was down-regulated in skeletal muscle of patients with genetically determined insulin resistance. Skeletal muscle biopsies......Transcriptional profiling of skeletal muscle from patients with type 2 diabetes and high-risk individuals have demonstrated a co-ordinated down-regulation of oxidative phosphorylation (OxPhos) genes, suggesting a link between insulin resistance and mitochondrial dysfunction. However, whether...... obtained during a euglycemic-hyperinsulinemic clamp in heterozygote carriers (n=6) of a mutation (Arg1174Gln) in the tyrosine kinase domain of the insulin receptor gene (INSR) and matched healthy controls (n=11) were used for transcriptional profiling. Biological pathways analysis was performed using Gene...

  8. Dual Amylin and Calcitonin Receptor Agonists: A Novel Treatment for Obesity and Related Co-Morbidities

    DEFF Research Database (Denmark)

    Gydesen, Sofie

    accumulation was eliminated by KBP treatment, and interestingly, KBPs alleviated hyperinsulinemia and improved glucose tolerance even with significantly lower insulin levels. KBP treatment increased the glucose infusion rate during a hyperinsulinemic euglycemic clamp indicating enhanced insulin action....... Importantly, KBPs also improved glucose homeostasis and enhanced insulin action in Zucker Diabetic Fatty rats. To investigate beneficial effects beyond weight loss, a weight-matched group was implemented. Of interest, weight matching led to improved glucose homeostasis through lowered plasma insulin; however...... the combination resulted in a 12% body weight reduction. Moreover, the combination improved glucose tolerance. In a rat model resembling the phenotype of human NASH, KBP treatment led to a reduction of the high fat, high cholesterol and cholate diet induced increase in liver weight and circulating aspartate...

  9. Changes in HbA1c and circulating and adipose tissue androgen levels in overweight-obese women with polycystic ovary syndrome in response to electroacupuncture

    DEFF Research Database (Denmark)

    Stener-Victorin, E.; Maliqueo, M.; Soligo, M

    2016-01-01

    AIM: Insulin sensitivity is ~40% lower in women with polycystic ovary syndrome (PCOS) than in controls. We tested the hypothesis that 5 weeks of electroacupuncture treatment improves glucose regulation and androgen levels in overweight/obese women with PCOS. MATERIAL AND METHODS: Seventeen women...... with PCOS, aged 18 to 38 years, with a body mass index (BMI) ≥25 kg/m(2) and diagnosed with PCOS were included in this experimental and feasibility study and subjected to five weeks of electroacupuncture treatments three times/week. The primary outcome was changes in whole-body glucose homeostasis measured...... by euglycemic hyperinsulinemic clamp before and after the intervention. Secondary outcome were changes in HbA1c, circulating catecholamines, adipocyte size and adipose tissue expression of sex steroids and nerve growth factor (NGF). RESULTS: No significant change in glucose homeostasis was observed, but HbA1c...

  10. Impact of TCF7L2 rs7903146 on insulin secretion and action in young and elderly Danish twins

    DEFF Research Database (Denmark)

    Wegner, Lise; Hussain, Meena S; Pilgaard, Kasper

    2008-01-01

    tests, and population 2 was additionally examined with iv glucose tolerance tests and hyperinsulinemic, euglycemic clamps. RESULTS: Elderly T-allele carriers had decreased plasma insulin responses and lower disposition index, whereas insulinogenic index was similar between genotype groups. Elderly...... capacity, birth, or adult anthropometry and was not associated with in vivo glucose metabolism. CONCLUSIONS: The rs7903146 T-allele associates with hepatic insulin resistance and diminished glucose-stimulated plasma insulin secretion. Our study does not provide evidence of a role of TCF7L2 gene expression...... in sc fat tissue and muscle tissue in the regulation of glucose homeostasis. This suggests that the primary defect of rs7903146 T-allele carriers is impairment of insulin secretion rather than a defect in insulin action in peripheral tissues....

  11. Increased subsarcolemmal lipids in type 2 diabetes: effect of training on localization of lipids, mitochondria, and glycogen in sedentary human skeletal muscle

    DEFF Research Database (Denmark)

    Nielsen, Joachim; Hey-Mogensen, Martin; Vind, Birgitte F

    2010-01-01

    The purpose of the study was to investigate the effect of aerobic training and type 2 diabetes on intramyocellular localization of lipids, mitochondria, and glycogen. Obese type 2 diabetic patients (n = 12) and matched obese controls (n = 12) participated in aerobic cycling training for 10 wk....... Endurance-trained athletes (n = 15) were included for comparison. Insulin action was determined by euglycemic-hyperinsulinemic clamp. Intramyocellular contents of lipids, mitochondria, and glycogen at different subcellular compartments were assessed by transmission electron microscopy in biopsies obtained...... from vastus lateralis muscle. Type 2 diabetic patients were more insulin resistant than obese controls and had threefold higher volume of subsarcolemmal (SS) lipids compared with obese controls and endurance-trained subjects. No difference was found in intermyofibrillar lipids. Importantly, following...

  12. Beta-cell function is associated with carotid intima-media thickness independently of insulin resistance in healthy individuals

    DEFF Research Database (Denmark)

    Roussel, Ronan; Natali, Andrea; Balkau, Beverley

    2016-01-01

    Objective: It is a common belief that early atherosclerosis in prediabetes is causally linked to endothelial insulin resistance. Another condition, a low insulin secretion, may be associated with insufficient insulin action on the vascular wall and consequently favor atherosclerosis. Our aim...... was to test this hypothesis in people without diabetes, taking into account the gold-standard measurement of insulin sensitivity, a major confounder in the relationship between insulin secretion and atherosclerosis. Methods: We studied the European Relationship between Insulin Sensitivity and Cardiovascular...... Risk cohort of 451 men and 593 women (44±8 years, mean±SD) who were free of diabetes, hypertension, dyslipidemia, and other known chronic or acute conditions. All underwent an oral glucose tolerance test, a euglycemic-hyperinsulinemic clamp (M/I measured insulin sensitivity), and B-mode carotid...

  13. Enhanced insulin signaling in human skeletal muscle and adipose tissue following gastric bypass surgery

    DEFF Research Database (Denmark)

    Albers, Peter Hjorth; Bojsen-Moller, Kirstine N; Dirksen, Carsten

    2015-01-01

    12 months post-surgery. Adipose tissue from glucose tolerant subjects was the most responsive to RYGB compared to type 2 diabetic patients, whereas changes in skeletal muscle were largely similar in these two groups. In conclusion, an improved molecular insulin sensitive phenotype of skeletal muscle......Roux-en-Y gastric bypass (RYGB) leads to increased peripheral insulin sensitivity. The aim of this study was to investigate the effect of RYGB on expression and regulation of proteins involved in regulation of peripheral glucose metabolism. Skeletal muscle and adipose tissue biopsies from glucose...... tolerant and type 2 diabetic subjects at fasting and during a hyperinsulinemic-euglycemic clamp before as well as 1 week, 3 and 12 months after RYGB were analyzed for relevant insulin effector proteins/signaling components. Improvement in peripheral insulin sensitivity mainly occurred at 12 months post-surgery...

  14. Higher intramuscular triacylglycerol in women does not impair insulin sensitivity and proximal insulin signaling

    DEFF Research Database (Denmark)

    Høeg, Louise; Roepstorff, Carsten; Thiele, Maja

    2009-01-01

    whole body (pinsulin stimulated leg glucose uptake (p=0.05) during an euglycemic-hyperinsulinemic ( approximately 70 uU/ml) clamp compared to matched male subjects. The higher insulin sensitivity in women could not be explained by higher expression of muscle GLUT4, insulin receptor (IR......Women have been shown to have higher muscle tricylglycerol (IMTG) levels than men and could therefore be expected to have lower insulin sensitivity than men, since previous studies have linked high IMTG to decreased insulin sensitivity. Thus, insulin sensitivity of whole body and leg glucose uptake......) or Akt expression or the ability of insulin to stimulate Akt (thr308) or Akt (ser473) phosphorylation. However, a 30% higher (p

  15. Young, low-birth-weight men are not more susceptible to the diabetogenic effects of a prolonged free fatty acid exposure than matched controls

    DEFF Research Database (Denmark)

    Jensen, Christine B; Storgaard, Heidi; Holst, Jens Juul

    2005-01-01

    Low birth weight (LBW) is associated with increased risk of developing type 2 diabetes later in life. Progression from normal to impaired glucose tolerance and overt diabetes may depend, to some extent, on elevation of plasma free fatty acids (FFAs). We undertook this study to elucidate whether...... a prolonged physiological lipid load could unmask or augment existing metabolic defects in otherwise healthy young LBW subjects. Forty 19-year-old men (LBW [n = 20], controls [normal birth weight, NBW] [n = 20]) without a family history of diabetes underwent an intravenous glucose tolerance test (0.3 g kg(-1......)), followed by 2-step hyperinsulinemic-euglycemic clamps (2 x 120 minutes: 10 and 40 mU m(-2) min(-1)) in combination with [3-3H]-glucose and indirect calorimetry. The tests were preceded, in randomized order, by a 24-hour continuous intralipid (20%, 0.4 mg mL(-1) h(-1)) or saline infusion. Estimates...

  16. Direct evidence of fiber type-dependent GLUT-4 expression in human skeletal muscle

    DEFF Research Database (Denmark)

    Gaster, M; Poulsen, P; Handberg, A

    2000-01-01

    GLUT-4 expression in individual fibers of human skeletal muscles in younger and older adults was studied. Furthermore, the dependency of insulin-stimulated glucose uptake on fiber type distribution was investigated. Fiber type distribution was determined in cryosections of muscle biopsies from 8...... younger (29 yr) and 8 older (64 yr) healthy subjects, and estimates of GLUT-4 expression in individual fibers were obtained by combining immunohistochemistry and stereology. GLUT-4 was more abundantly expressed in slow compared with fast muscle fibers in both younger (P ...) subjects. A 25% reduction of GLUT-4 density in fast fibers (P GLUT-4 density in slow fibers were demonstrated in older compared with younger subjects. Insulin-stimulated glucose uptake rates measured by hyperinsulinemic, euglycemic clamp were not correlated with the fraction...

  17. Direct effects of locally administered lipopolysaccharide on glucose, lipid, and protein metabolism in the placebo-controlled, bilaterally infused human leg

    DEFF Research Database (Denmark)

    Buhl, Mads; Bosnjak, Ermina; Vendelbo, Mikkel H.

    2013-01-01

    discrimination between direct and indirect effects impossible. Objective: We sought to define the direct, placebo-controlled effects of LPS on insulin resistance and protein and lipid metabolism in the infused human leg without systemic interference from cytokines and stress hormones. Design......Context: Accumulating evidence suggests that chronic exposure to lipopolysaccharide (LPS, endotoxin) maycreate a constant low-grade inflammation, leading to insulin resistance and diabetes. All previous human studies assessing the metabolic actions of LPS have used systemic administration, making......: This was a randomized, placebo-controlled, single-blinded study. Participants and Intervention: We studied 8 healthy volunteers with bilateral femoral vein and artery catheters during a 3-hour basal and 3-hour hyperinsulinemic-euglycemic clamp period with bilateral muscle biopsies in each period during infusion...

  18. Effects of 7 days of exercise training on insulin sensitivity and responsiveness in type 2 diabetes mellitus

    DEFF Research Database (Denmark)

    Kirwan, John P; Solomon, Thomas; Wojta, Daniel M

    2009-01-01

    The objectives of this study were to determine whether 1) the improvement in insulin action induced by short-term exercise training in patients with type 2 diabetes is due to an improvement in insulin sensitivity, an improvement in insulin responsiveness, or a combination of improved insulin...... sensitivity and responsiveness and 2) short-term exercise training results in improved suppression of hepatic glucose production by insulin. Fourteen obese patients with type 2 diabetes, age 64 +/- 2 yr, underwent a two-stage hyperinsulinemic euglycemic clamp procedure, first stage 40 mU.m(-2).min(-1) insulin...... of vigorous exercise training can induce significant improvements in insulin action in type 2 diabetes. These improvements include increased peripheral insulin sensitivity and responsiveness as well as enhanced suppression of hepatic glucose production....

  19. Hydrochlorothiazide, but not Candesartan, aggravates insulin resistance and causes visceral and hepatic fat accumulation: the mechanisms for the diabetes preventing effect of Candesartan (MEDICA) Study

    DEFF Research Database (Denmark)

    Eriksson, J.W.; Jansson, P.A.; Carlberg, B.

    2008-01-01

    Treatment with angiotensin II receptor blockers is associated with lower risk for the development of type 2 diabetes mellitus compared with thiazide diuretics. The Mechanisms for the Diabetes Preventing Effect of Candesartan Study addressed insulin action and secretion and body fat distribution...... after treatment with candesartan, hydrochlorothiazide, and placebo. Twenty-six nondiabetic, abdominally obese, hypertensive patients were included in a multicenter 3-way crossover trial, and 22 completers (by predefined criteria; 10 men and 12 women) were included in the analyses. They underwent 12-week...... treatment periods with candesartan (C; 16 to 32 mg), hydrochlorothiazide (H; 25 to 50 mg), and placebo (P), respectively, and the treatment order was randomly assigned and double blinded. Intravenous glucose tolerance tests and euglycemic hyperinsulinemic (56 mU/m(2) per minute) clamps were performed...

  20. IRS-1 serine phosphorylation and insulin resistance in skeletal muscle from pancreas transplant recipients

    DEFF Research Database (Denmark)

    Bouzakri, Karim; Karlsson, Håkan K R; Vestergaard, Henrik

    2006-01-01

    Insulin-dependent diabetic recipients of successful pancreas allografts achieve self-regulatory insulin secretion and discontinue exogenous insulin therapy; however, chronic hyperinsulinemia and impaired insulin sensitivity generally develop. To determine whether insulin resistance is accompanied...... by altered signal transduction, skeletal muscle biopsies were obtained from pancreas-kidney transplant recipients (n = 4), nondiabetic kidney transplant recipients (receiving the same immunosuppressive drugs; n = 5), and healthy subjects (n = 6) before and during a euglycemic-hyperinsulinemic clamp. Basal...... insulin receptor substrate (IRS)-1 Ser (312) and Ser (616) phosphorylation, IRS-1-associated phosphatidylinositol 3-kinase activity, and extracellular signal-regulated kinase (ERK)-1/2 phosphorylation were elevated in pancreas-kidney transplant recipients, coincident with fasting hyperinsulinemia. Basal...

  1. Effects of menopause and high-intensity training on insulin sensitivity and muscle metabolism

    DEFF Research Database (Denmark)

    Mandrup, Camilla M; Egelund, Jon; Nyberg, Michael

    2018-01-01

    To investigate peripheral insulin sensitivity and skeletal muscle glucose metabolism in premenopausal and postmenopausal women, and evaluate whether exercise training benefits are maintained after menopause. Sedentary, healthy, normal-weight, late premenopausal (n = 21), and early postmenopausal (n...... = 20) women were included in a 3-month high-intensity exercise training intervention. Body composition was assessed by magnetic resonance imaging and dual-energy x-ray absorptiometry, whole body glucose disposal rate (GDR) by hyperinsulinemic euglycemic clamp (40 mU/m/min), and femoral muscle glucose....... Training increased lean body mass (estimate [95% confidence interval] 0.5 [0.2-0.9] kg, P ...

  2. Off-clamp robot-assisted partial nephrectomy does not benefit short-term renal function: a matched cohort analysis.

    Science.gov (United States)

    Anderson, Barrett G; Potretzke, Aaron M; Du, Kefu; Vetter, Joel; Figenshau, R Sherburne

    2017-08-31

    In the interest of renal functional preservation, partial nephrectomy has supplanted radical nephrectomy as the preferred treatment for T1 renal masses. This procedure usually involves the induction of renal warm ischemia by clamping the hilar vessels prior to tumor excision. Performing robot-assisted partial nephrectomy (RAPN) "off-clamp" can theoretically prevent renal functional loss associated with warm ischemia. We describe our institutional experience and compare perioperative and renal functional outcomes using a propensity score matched cohort. We conducted a retrospective comparison from a prospectively maintained database of all patients who underwent RAPN from 2009 to 2015. Of those patients, 143 underwent off-clamp RAPN. Fifty off-clamp RAPN patients were propensity score matched with fifty clamped RAPN patients based on renal function, tumor size, and R.E.N.A.L. nephrometry score. The cohorts were compared across demographics, operative information, perioperative outcomes, and renal functional outcomes. For all off-clamp RAPN patients, mean nephrometry score was 7.1, mean estimated blood loss (EBL) was 236.9 mL, perioperative complication rate was 7.7%, and mean decrease in estimated glomerular filtration rate (eGFR) was 7.1% at a median follow-up of 9.2 months. In the propensity score matched cohorts, off-clamp RAPN resulted in a shorter mean operative time (172.0 versus 196.0 min, p = 0.025) and a lower mean EBL (179.7 versus 283.2 mL, p = 0.046). A lower complication rate of 6.0% in the off-clamp group compared with 20.0% in the clamped group approached significance (p = 0.071). Mean preoperative eGFR was similar in both cohorts. Importantly, there was no significant difference in decrease in eGFR between the clamped cohort (9.8%) and off-clamp cohort (11.9%) at a median follow-up of 9.0 months (p = 0.620). Off-clamp RAPN did not result in improved renal functional preservation in our experience. Surprisingly, the off-clamp cohort

  3. The Effects of Telmisartan and Losartan on Insulin Sensitivity

    Directory of Open Access Journals (Sweden)

    Okan Bakıner

    2013-12-01

    Full Text Available Purpose: Telmisartan has been reported to increase insulin sensitivity by acting as a partial PPAR-gamma agonist, regardless of its renin-angiotensin system inhibition, but losartan has no such activity. In this study, we compared the effects of telmisartan and losartan on insulin sensitivity among type 2 diabetic patients. Material and Method: Age-, sex- and weight-matched patients, who had been on telmisartan or losartan treatment for at least 3 months, were included. Their anthropometric measurements were performed, blood pressures were recorded. Fasting venous blood samples were obtained for analyzing the levels of glucose, HbA1C, insulin and adiponectin. HOMA-IR was calculated. An euglycemic hyperinsulinemic clamp procedure was performed in each subject and M index was calculated. Thereafter, telmisartan and losartan were withdrawn in both groups. A cross-over design was planned; following a wash-out period of 15 days, losartan group (Group 1 was given telmisartan 80 mg/day, telmisartan group (Group 2 was administered losartan 50 mg/day. After 12 weeks of therapy, all measurements, calculations and the euglycemic hyperinsulinemic clamp test were re-performed. Results: General characteristics of the patients at inclusion were similar. Nine cases in group 1 and 8 cases in Group 2 concluded the follow-up. In Group 1, among all follow-up parameters, only weight exhibited a significant decrease at final evaluation (p=0,034. In Group 2, only M value was found to increase (p=0,028. Discussion: Our findings indicated that losartan improved insulin sensitivity in patients with concomitant hypertension and type 2 diabetes, and telmisartan use resulted in more weight loss. Turk Jem 2013; 17: 92-7

  4. Increased fetal myocardial sensitivity to insulin-stimulated glucose metabolism during ovine fetal growth restriction.

    Science.gov (United States)

    Barry, James S; Rozance, Paul J; Brown, Laura D; Anthony, Russell V; Thornburg, Kent L; Hay, William W

    2016-04-01

    Unlike other visceral organs, myocardial weight is maintained in relation to fetal body weight in intrauterine growth restriction (IUGR) fetal sheep despite hypoinsulinemia and global nutrient restriction. We designed experiments in fetal sheep with placental insufficiency and restricted growth to determine basal and insulin-stimulated myocardial glucose and oxygen metabolism and test the hypothesis that myocardial insulin sensitivity would be increased in the IUGR heart. IUGR was induced by maternal hyperthermia during gestation. Control (C) and IUGR fetal myocardial metabolism were measured at baseline and under acute hyperinsulinemic/euglycemic clamp conditions at 128-132 days gestation using fluorescent microspheres to determine myocardial blood flow. Fetal body and heart weights were reduced by 33% (P = 0.008) and 30% (P = 0.027), respectively. Heart weight to body weight ratios were not different. Basal left ventricular (LV) myocardial blood flow per gram of LV tissue was maintained in IUGR fetuses compared to controls. Insulin increased LV myocardial blood flow by ∼38% (P IUGR fetuses was 73% greater than controls. Similar to previous reports testing acute hypoxia, LV blood flow was inversely related to arterial oxygen concentration (r(2 )= 0.71) in both control and IUGR animals. Basal LV myocardial glucose delivery and uptake rates were not different between IUGR and control fetuses. Insulin increased LV myocardial glucose delivery (by 40%) and uptake (by 78%) (P IUGR fetuses compared to controls. During basal and hyperinsulinemic-euglycemic clamp conditions LV myocardial oxygen delivery, oxygen uptake, and oxygen extraction efficiency were not different between groups. These novel results demonstrate that the fetal heart exposed to nutrient and oxygen deprivation from placental insufficiency appears to maintain myocardial energy supply in the IUGR condition via increased glucose uptake and metabolic response to insulin, which support

  5. Dietary fat drives whole-body insulin resistance and promotes intestinal inflammation independent of body weight gain.

    Science.gov (United States)

    Jensen, Benjamin A H; Nielsen, Thomas S; Fritzen, Andreas M; Holm, Jacob B; Fjære, Even; Serup, Annette K; Borkowski, Kamil; Risis, Steve; Pærregaard, Simone I; Søgaard, Ida; Poupeau, Audrey; Poulsen, Michelle; Ma, Tao; Sina, Christian; Kiens, Bente; Madsen, Lise; Kristiansen, Karsten; Treebak, Jonas T

    2016-12-01

    The obesogenic potential of high-fat diets (HFD) in rodents is attenuated when the protein:carbohydrate ratio is increased. However, it is not known if intake of an HFD irrespective of the protein:carbohydrate ratio and in the absence of weight gain, affects glucose homeostasis and the gut microbiota. We fed C57BL6/J mice 3 different HFDs with decreasing protein:carbohydrate ratios for 8weeks and compared the results to a LFD reference group. We analyzed the gut microbiota composition by 16S rDNA amplicon sequencing and the intestinal gene expression by real-time PCR. Whole body glucose homeostasis was evaluated by insulin and glucose tolerance tests as well as by a hyperinsulinemic euglycemic clamp experiment. Compared with LFD-fed reference mice, HFD-fed mice, irrespective of protein:carbohydrate ratio, exhibited impaired glucose tolerance, whereas no differences were observed during insulin tolerance tests. The hyperinsulinemic euglycemic clamp revealed tissue-specific effects on glucose homeostasis in all HFD-fed groups. HFD-fed mice exhibited decreased insulin-stimulated glucose uptake in white but not in brown adipose tissue, and sustained endogenous glucose production under insulin-stimulated conditions. We observed no impairment of insulin-stimulated glucose uptake in skeletal muscles of different fiber type composition. HFD-feeding altered the gut microbiota composition paralleled by increased expression of pro-inflammatory cytokines and genes involved in gluconeogenesis in intestinal epithelial cells of the jejunum. Intake of a HFD profoundly affected glucose homeostasis, gut inflammatory responses, and gut microbiota composition in the absence of fat mass accretion. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  6. NMR resonance assignments for the N-terminal domain of the δ subunit of the E. coli γ clamp loader complex.

    Science.gov (United States)

    Alyami, Esmael M; Rizzo, Alessandro A; Beuning, Penny J; Korzhnev, Dmitry M

    2017-10-01

    The β-clamp protein and the γ clamp loader complex are essential components of bacterial DNA replication machinery. The β-clamp is a ring-shaped homodimer that encircles DNA and increases the efficiency of replication by providing a binding platform for DNA polymerases and other replication-related proteins. The β-clamp is loaded onto DNA by the five-subunit γ clamp loader complex in a multi-step ATP-dependent process. The initial steps of this process involve the cooperative binding of the β-clamp by the five subunits of ATP-bound clamp loader, which induces or traps an open conformation of the clamp. Remarkably, the δ subunit of the E. coli clamp loader, or even its 140 residue N-terminal domain (called mini-δ), alone can shift conformational equilibrium of the β-clamp towards the open state. Here we report nearly complete backbone and side-chain 1H, 13C and 15N NMR resonance assignments of mini-δ that will facilitate NMR studies of the mechanisms of β-clamp opening and its loading on DNA by the clamp loader.

  7. Glucose effectiveness and insulin sensitivity measurements derived from the non-insulin-assisted minimal model and the clamp techniques are concordant

    DEFF Research Database (Denmark)

    Henriksen, Jan Erik; Alford, Frank; Ward, Glenn

    2010-01-01

    We investigated the concordance between glucose effectiveness (SG) and insulin sensitivity (SI), derived from the unmodified dynamic non-insulin-assisted intravenous glucose tolerance test (IVGTT) implemented by SG(MM) and SI(MM); simulation analysis and modelling/conversational interaction (SAAM....../CONSAM) versus the eu/hyperglycaemic basal insulinaemic and the euglycaemic hyperinsulinaemic clamp (SG(CLAMP) and SI(CLAMP))....

  8. Implementation and Tuning of an Optical Tweezers Force-Clamp Feedback System.

    Science.gov (United States)

    Bugiel, Michael; Jannasch, Anita; Schäffer, Erik

    2017-01-01

    Feedback systems can be used to control the value of a system variable. In optical tweezers, active feedback is often implemented to either keep the position or tension applied to a single biomolecule constant. Here, we describe the implementation of the latter: an optical force-clamp setup that can be used to study the motion of processive molecular motors under a constant load. We describe the basics of a software-implemented proportional-integral-derivative (PID) controller, how to tune it, and how to determine its optimal feedback rate. Limitations, possible feed-forward applications, and extensions into two- and three-dimensional optical force clamps are discussed. The feedback is ultimately limited by thermal fluctuations and the compliance of the involved molecules. To investigate a particular mechanical process, understanding the basics and limitations of the feedback system will be helpful for choosing the proper feedback hardware, for optimizing the system parameters, and for the design of the experiment.

  9. Analisa Sistem Clamp pada Penggantung Jembatan Kutai Kartanegara dengan Metode Elemen Hingga

    Directory of Open Access Journals (Sweden)

    Sri Nur Oktavianto

    2013-03-01

    Full Text Available Kegagalan suatu struktur dapat terjadi akibat perubahan dan terkonsentrasinya gaya-gaya dalam diluar prediksi pada saat perencanaan. Seperti halnya yang terjadi pada Jembatan Kutai Kertanegara (Kukar yang merupakan jembatan bertipe suspension terpanjang yang dimiliki oleh Indonesia, mengalami keruntuhan pada saat proses pemeliharaan berlangsung. Penyebab runtuhnya Jembatan Kukar diperkirakan rusaknya bagian koneksi clamp yang disebabkan oleh terkonsentrasinya gaya akibat proses jacking salah satu hanger pada saat proses penyesuaian camber jembatan, selain itu korosi dari material yang digunakan juga ikut andil dalam hal ini. Salah satu metode yang bisa digunakan adalah Finite Elemen Method (FEM, metode numerik ini digunakan untuk menyelesaikan permasalahan teknik dan problem matematis dari suatu gejala phisis[1]. Untuk membantu proses simulasi diperlukan program bantu yang berbasis FEM. Dari hasil simulasi didapat gaya yang bekerja pada saat mengalami keruntuhan dan konsentrasi tegangan pada sistem clamp.

  10. Requirements to gap widths and clamping for CO2 laser butt welding

    DEFF Research Database (Denmark)

    Gong, Hui; Juhl, Thomas Winther

    1999-01-01

    In the experimental study of fixturing and gap width requirements a clamping device for laser butt welding of steel sheets has been developed and tested. It has fulfilled the work and made the gap width experiments possible.It has shown that the maximum allowable gap width to some extent is inver......In the experimental study of fixturing and gap width requirements a clamping device for laser butt welding of steel sheets has been developed and tested. It has fulfilled the work and made the gap width experiments possible.It has shown that the maximum allowable gap width to some extent....../min 2.6 kWThe quality level is measured according to ISO 13919-1. Qualities of the butt welds with the maximum gap width listed in the table are mainly grouped in level B (stringent). The maximum gap width should be chosen with respect to the application requirements....

  11. Axially Modulated Clamped-Guided Arch Resonator for Memory and Logic Applications

    KAUST Repository

    Hafiz, Md Abdullah Al

    2017-11-03

    We experimentally demonstrate memory and logic devices based on an axially modulated clamped-guided arch resonator. The device are electrostatically actuated and capacitively sensed, while the resonance frequency modulation is achieved through an axial electrostatic force from the guided side of the clamped-guided arch microbeam. We present two case studies: first, a dynamic memory based on the nonlinear frequency response of the resonator, and second, a reprogrammable two-input logic gate based on the linear frequency modulation of the resonator. These devices show energy cost per memory/logic operation in pJ, are fully compatible with CMOS fabrication processes, have the potential for on-chip system integration, and operate at room temperature.

  12. [Traumatic section of the thoracic aorta: its repair by direct aortic clamping].

    Science.gov (United States)

    Cairols, M A; Sieyro, F; Miralles, M; Blanes, I; Lozano, P

    1991-01-01

    Traumatic section of descendent aorta, severe complication of a thoracic traumatism, requires an early recognition and restoration because its high mortality rate. Between 1988 and 1990, three patients underwent surgical approach, by direct aortic clamping [correction of clamplaje], at our Service. In two cases, a heart stoppage appeared during the clamping/unclamping [correction of clampaje/desclampaje] maneuvers. One of them had previously electrocardiographic abnormalities, suggestive of heart contusion. Two patients died, one of them during surgical procedure because an irreversible heart stoppage, and the second patient died after 7 days because of a brain hemorrhage. One case presented postoperative paraplegia. Respective rates and literature about the main factors implicated in the diagnosis and treatment of such pathology are reviewed.

  13. Voltage-clamp recordings from identified dissociated neuroendocrine cells of the adult rat supraoptic nucleus.

    Science.gov (United States)

    Cobbett, P; Weiss, M L

    1990-06-01

    In vitro intracellular recordings of membrane potential obtained from the oxytocin and vasopressin neurons of the mammalian hypothalamo-neurohypophysial system in slices (1-3) and expiants (4, 5) have demonstrated many of the intrinsic properties of these magnocellular neuroendocrine cells (MNCs). Voltage-clamp techniques, which are required to study directly the currents underlying intrinsic or transmitter-evoked potential changes, have been applied to cultured embryonic (6) or neonatal supraoptic neurons (7-9) and have been successfully applied to adult supraoptic neurons in situ in only one laboratory (10, 11). We have modified a technique for dissociation of viable adult guineapig hippocampal neurons (12) to dissociate supraoptic MNCs from adult rats for voltage-clamp studies. MNCs were selectively labelled with a fluorescent dye in vivo so that they could be identified after dissociation and prior to making recordings. These data have been published in abstract form elsewhere (13, 14).

  14. Five-Level Z-Source Neutral Point-Clamped Inverter

    DEFF Research Database (Denmark)

    Gao, F.; Loh, P.C.; Blaabjerg, Frede

    2007-01-01

    This paper proposes a five-level Z-source neutralpoint- clamped (NPC) inverter with two Z-source networks functioning as intermediate energy storages coupled between dc sources and NPC inverter circuitry. Analyzing the operational principles of Z-source network with partial dclink shoot-through s......This paper proposes a five-level Z-source neutralpoint- clamped (NPC) inverter with two Z-source networks functioning as intermediate energy storages coupled between dc sources and NPC inverter circuitry. Analyzing the operational principles of Z-source network with partial dclink shoot......-through scheme reveals the hidden theories in the five-level Z-source NPC inverter unlike the operational principle appeared in the general two-level Z-source inverter, so that the five-level Z-source NPC inverter can be designed with the modulation of carrier-based phase disposition (PD) or alternative phase...

  15. Indications, techniques, outcomes, and limitations for minimally ischemic and off-clamp partial nephrectomy: a systematic review of the literature.

    Science.gov (United States)

    Simone, Giuseppe; Gill, Inderbir S; Mottrie, Alexandre; Kutikov, Alexander; Patard, Jean-Jacques; Alcaraz, Antonio; Rogers, Craig G

    2015-10-01

    On-clamp partial nephrectomy (PN) has been considered the standard approach to minimize intraoperative bleeding and thus achieve adequate control of tumor margins. The potential negative impact of ischemia on renal function (RF) led to the development of techniques to minimize or avoid renal ischemia, such as off-clamp PN and minimally ischemic PN techniques. To review current evidence on the indications and techniques for and outcomes of minimally ischemic and off-clamp PN. A systematic review of English-language publications on PN without a main renal artery clamp from January 2005 to July 2014 was performed using the Medline, Embase, and Web of Science databases. The searches retrieved 52 papers. Off-clamp PN has been more commonly applied to small and peripheral renal tumors, while minimally ischemic PN is best suited for hilar and medially located renal tumors. These approaches are associated with increased intraoperative blood loss and perioperative transfusion rates compared to on-clamp PN. Minimally ischemic and off-clamp PN have potential functional benefits when longer ischemia time is anticipated, particularly for patients with lower baseline RF. Limitations include the lack of prospective randomized trials comparing minimally ischemic and off-clamp to on-clamp techniques, and the small sample size and short follow-up of most published series. The impact of different resection and renorrhaphy techniques on postoperative RF and its assessment via renal scintigraphy requires further investigations. Minimally ischemic and off-clamp PN are established procedures that may be particularly applicable for patients with decreased baseline RF. However, these techniques are technically demanding, with potential for increased blood loss, and require considerable experience with PN surgery. The role of ischemia in patients with a contralateral healthy kidney and consequently an indication for elective minimally ischemic or off-clamp PN remains a debatable issue. In

  16. Reversible fastener clamp load monitor with continuous visual or remote readout

    Science.gov (United States)

    Hodge, Malcolm H.; Kausel, Theodore C., Jr.; Begley, Matthew R.

    1998-03-01

    SIMS has developed a simple means for detecting and monitoring both absolute and relative clamp load, or bolt tension, in fastener systems. More than twenty-five percent of automotive failures are known to be due to undetected loss of fastener clamp load. While the equivalent aerospace maintenance statistics are not known, the average automobile has 3,500 fasteners while a Boeing 747 has closer to one million. It is therefore anticipated that the new SensaBolt clamp load tracking system could find wide applications in the aerospace arena. We describe a visually-evident and retrofitted clamp load monitoring design which is based on the differential joint substrate compression at, and immediately adjacent to, the fastener location. This intrinsically-accurate indicator does not necessarily require alteration in either the bolt or nut geometries, thereby facilitating product introduction and retrofit in aging aircraft applications. In addition, SensaBolt's sole reliance on substrate compression renders it more accurate then torque wrench or turn-of-nut techniques. Readout may be accomplished by any of three principal methods: for those applications with ease of access to the sensor, loss of tension can be determined by direct visual inspection. Application of a standard wrench can then be made to restore the fastener's proper tightness, per the SensaBolt indicators. In those instances where line-of-sight is unimpeded and more formal inspection is desired, the SensaBolt may be interrogated by a laser scanner bar code reader. Finally, SensaBolt may be addressed by the SIMS fiber optic harness for those instances where full-time remote interrogation is desired.

  17. Adverse impact of intermittent portal clamping on long-term postoperative outcomes in hepatocellular carcinoma.

    Science.gov (United States)

    Hao, S; Chen, S; Yang, X; Wan, C

    2017-01-01

    Introduction To evaluate the impact of intermittent portal clamping (IPC) on long-term postoperative outcomes in patients with hepatocellular carcinoma (HCC). Methods Clinical records of 355 patients underwent curative liver resection for HCC in January 2007 to December 2010 were retrospectively reviewed. According to how portal clamping was performed, patients were grouped as: IPC, n=113; other portal clamping (OPC), n=190; and no portal clamping (NPC), n=52. Results Median recurrence-free survival (RFS) was statistically significantly shorter in the IPC (39.4 months) than OPC (47.3 months, p=0.010) and NPC groups (51.4 months, p=0.008). Median overall survival (OS) was also significantly shorter with IPC (46.3 months), versus 52.9 months with OPC (p=0.022) and 56.2 months with NPC (p=0.015). Kaplan-Meier survival analysis revealed that 5-year cumulative RFS was much lower in the IPC (42.5%) than OPC (50.9%, p=0.014) and NPC groups (49.6%, p=0.013). Five-year cumulative OS was also much lower in the IPC (44.9%) than OPC (58.0%, p=0.020) and NPC groups (57.7%, p=0.025). On univariate analysis, tumour grade, size and number, TNM stage, blood transfusion, vascular invasion and IPC were significantly inversely correlated with RFS and OS. On multivariate analysis, tumour size and number, blood transfusion, vascular invasion and IPC remained significant. Conclusions Our study suggests that IPC is an independent risk factor for poor long-term postoperative outcomes in patients with HCC.

  18. Clamp fixation to prevent unfolding of a suture knot decreases tensile strength of polypropylene sutures.

    Science.gov (United States)

    Türker, Mehmet; Yalçinozan, Mehmet; Çirpar, Meriç; Çetik, Özgür; Kalaycioğlu, Barış

    2012-12-01

    Although sutures evolved in last decade and the product spectrum broadened largely, they can be still classified into two: monofilament and multifilament. Sutures are the mainstay of orthopedic procedures like fascial closures, tendon repairs or tenodesis. In every repair, a suture loop is created. This suture loop is prone to failure due to suture elongation, knot slip and suture breakage. As the knot is the stress riser in a suture loop, the majority of acute loop failure occurs just adjacent to the knot. Monofilament sutures have higher bending stiffness and tendency to untie than multifilament sutures. The first throw of monofilament sutures have tendency to untie, which decrease loop tension and result in loss of achieved tissue approximation. Although a common practice is to fix the first throw via a clamp before the locking one is tied, it can be hypothesized that a potential deforming effect can lead to a decrease in ultimate failure load of a monofilament suture loop. Fixing the first throw significantly reduced the ultimate failure load of monofilament nonabsorbable polypropylene sutures (Prolene) (62.2 ± 8 N vs. 72.7 ± 9 N, p = 0.019). The ultimate failure load achieved by monofilament sutures Polyglyconate (Maxon) and Nylon (Ethilon) and braided absorbable Polyglactin (Vicryl) were not affected by fixing the first throw. Under microscopic examination, polypropylene sutures were found to be deformed by clamp fixation, while the others were not. Polypropylene sutures can be easily damaged when it is fixed by a clamp during knot tying. Presented data demonstrated that in real surgical situations clamp fixation of polypropylene knots can damage the suture loop and carry the risk of acute failure of repair site during early rehabilitation.

  19. Calcium-activated potassium current clamps the dark potential of vertebrate rods.

    Science.gov (United States)

    Moriondo, A; Pelucchi, B; Rispoli, G

    2001-07-01

    Vertebrate photoreceptors respond to light with a graded hyperpolarization from a membrane potential in the dark of approximately -35 mV. The present work investigates the physiological role of the Ca2+-activated K+ current in the photovoltage generation in mechanically isolated rods from salamander retina. Membrane current or voltage in isolated rods was recorded from light- and dark-adapted rods under voltage- or current-clamp conditions, respectively. In light-adapted rods of the salamander, selective blockade of Ca2+-activated K+ channels by means of charybdotoxin depolarized the plasma membrane of current-clamped rods by approximately 30 mV, from a resting potential of approximately -35 mV. A similar depolarization was observed if external Ca2+ (1 mM) was substituted with Ba2+ or Sr2+. Under control conditions, the injection of currents of increasing amplitude (up to -100 pA, to mimic the current entering the rod outer segment) could not depolarize the membrane potential beyond a saturating value of approximately -20 mV. However, in the presence of charybdotoxin, rods depolarized up to +20 mV. In experiments with dark-adapted current-clamped rods, charybdotoxin perfusion lead to transient depolarizations up to 0 mV and steady-state depolarizations of approximately 5 mV above the dark resting potential. Finally, the recovery phase of the voltage response to a flash of light in the presence of charybdotoxin showed a transient overshoot of the membrane potential. It was concluded that Ca2+-activated K+ current is necessary for clamping the rod photovoltage to values close to the dark potential, thus allowing faithful single photon detection and correct synaptic transmission.

  20. Prediction of numerical distortion after welding with various welding sequences and clampings

    Directory of Open Access Journals (Sweden)

    S. Kastelic

    2010-10-01

    Full Text Available Welding simulation of a test cover for hydropower plant was made due to very large dimensions of the cover. The main aim was to predict distortion after welding in order to avoid machining the cover. Welding process was simulated with the Sysweld program to keep distortion in desired limits. Various welding sequences and clamping conditions were calculated to reduce the distortion. Calculation of microstructure constituents in virtual complex geometry of joints was also analyzed.

  1. Involvement of the β Clamp in Methyl-directed Mismatch Repair in Vitro*

    Science.gov (United States)

    Pluciennik, Anna; Burdett, Vickers; Lukianova, Olga; O'Donnell, Mike; Modrich, Paul

    2009-01-01

    We have examined function of the bacterial β replication clamp in the different steps of methyl-directed DNA mismatch repair. The mismatch-, MutS-, and MutL-dependent activation of MutH is unaffected by the presence or orientation of loaded β clamp on either 3′ or 5′ heteroduplexes. Similarly, β is not required for 3′ or 5′ mismatch-provoked excision when scored in the presence of γ complex or in the presence of γ complex and DNA polymerase III core components. However, mismatch repair does not occur in the absence of β, an effect we attribute to a requirement for the clamp in the repair DNA synthesis step of the reaction. We have confirmed previous findings that β clamp interacts specifically with MutS and MutL (López de Saro, F. J., Marinus, M. G., Modrich, P., and O'Donnell, M. (2006) J. Biol. Chem. 281, 14340–14349) and show that the mutator phenotype conferred by amino acid substitution within the MutS N-terminal β-interaction motif is the probable result of instability coupled with reduced activity in multiple steps of the repair reaction. In addition, we have found that the DNA polymerase III α catalytic subunit interacts strongly and specifically with both MutS and MutL. Because interactions of polymerase III holoenzyme components with MutS and MutL appear to be of limited import during the initiation and excision steps of mismatch correction, we suggest that their significance might lie in the control of replication fork events in response to the sensing of DNA lesions by the repair system. PMID:19783657

  2. Limitations of the whole cell patch clamp technique in the control of intracellular concentrations.

    OpenAIRE

    Mathias, R T; Cohen, I.S.; Oliva, C.

    1990-01-01

    Recent experimental studies (Pusch and Neher, 1988) and theoretical studies (Oliva et al., 1988) have found that the pipette tip is a significant barrier to diffusion in the whole cell patch clamp configuration. In this paper, we extend the theoretical analysis of fluxes between the pipette and cell to include transmembrane fluxes. The general conclusions are: (a) within the pipette, ion fluxes are driven primarily by diffusion rather than voltage gradients. (b) At steady state there is a con...

  3. Calculation of time constants for intracellular diffusion in whole cell patch clamp configuration

    OpenAIRE

    Oliva, C.; Cohen, I.S.; Mathias, R T

    1988-01-01

    We present a simplified model to identify and analyze the important variables governing the diffusion of substances from pipettes into canine cardiac Purkinje cells in the whole cell patch clamp configuration. We show that diffusion of substances through the pipette is the major barrier for equilibration of the pipette and cellular contents. We solve numerically the one-dimensional diffusion equation for different pipette geometries, and we derive a simple analytic equation which allows one t...

  4. Pengembangan Topologi Inverter Multilevel Tiga Tingkat Satu Fasa Tipe Dioda Clamped Dengan Mereduksi Komponen Saklar Daya

    OpenAIRE

    Matalata, Hendi; Hamid, Muh. Imran

    2016-01-01

    Multilevel inverter is a converter which converts the DC power source into an AC power source with a voltage output of more than two levels. Conventional multilevel inverter topology for the three levels developed at this time generally uses four components of the power switch. In this paper presented a three-level multilevel inverter topology by reducing the power switch components into two pieces and named as the type of diode clamped multilevel inverter three level one phase. Simulation wi...

  5. Prospective study comparing two techniques of renal clamping in laparoscopic partial nephrectomy: impact on perioperative parameters.

    Science.gov (United States)

    Imbeault, Annie; Pouliot, Frédéric; Finley, David S; Shuch, Brian; Dujardin, Thierry

    2012-05-01

    To compare en bloc and artery-only clamping techniques on renal function and perioperative outcomes after laparoscopic partial nephrectomy (LPN). From March 2003 to December 2008, 205 patients underwent LPN by one surgeon in a single institution. The first 103 LPNs were achieved with artery-only clamping (AO), and the last 102 LPNs were realized under control of the renal hilum (artery and vein [AV] clamping). Renal function was evaluated by creatinine changes, estimation of the glomerular filtration rate (eGFR), and assessment of split renal function using renal mercaptoacetyl triglycine-Lasix scintigraphy. Sixty-two of 205 patients had renal scintigraphy before and after surgery. There was no significant difference between the two groups regarding demographic data and renal mass characteristics. Warm ischemia time (WIT) was higher in the AO group: 30.4 ± 8.2 vs 23.3 minutes ± 10.0 (P<0.0001). The eGFR change was significantly lower in the AV group during the postoperative period: 10.2 mL/min vs 13.7 mL/min (P=0.0472). Operative blood loss, operative time, and complication rate were not statistically different between groups. Average loss of differential function of the operated kidney was 13.6 ± 9.2% for the AO group and 14.3 ± 12.3% for the AV group (P=0.8016). On multivariate analysis, clamping technique was not a predictive factor of renal function reduction. AV and AO techniques are associated with similar renal function outcomes in patients who were undergoing LPN. In our series, the AV technique was associated with a lower WIT, an important predictor of decrease in renal function.

  6. Crystal structure of an archaeal DNA sliding clamp: proliferating cell nuclear antigen from Pyrococcus furiosus.

    Science.gov (United States)

    Matsumiya, S; Ishino, Y; Morikawa, K

    2001-01-01

    The proliferating cell nuclear antigen (PCNA) is now recognized as one of the key proteins in DNA metabolic events because of its direct interactions with many proteins involved in important cellular processes. We have determined the crystal structure of PCNA from a hyperthermophilic archaeon, Pyrococcus furiosus (pfuPCNA), at 2.1 A resolution. pfuPCNA forms a toroidal, ring-shaped structure consisting of homotrimeric molecules, which is also observed in the PCNA crystals from human and yeast. The overall structure of pfuPCNA is highly conserved with other PCNA proteins, as well as with the bacterial ss clamp and the bacteriophage gp45. This result shows that the three-dimensional structure of the sliding clamp is conserved in the three domains of life. pfuPCNA has two remarkable features compared with the human and yeast PCNA molecules: it has more ion pairs and fewer intermolecular main chain hydrogen bonds. The former may contribute to the thermal stability of pfuPCNA, and the latter may be the cause of the stimulatory effect of pfuPCNA on the DNA synthesizing activity of P. furiosus DNA polymerases in the absence of the clamp loader replication factor C in vitro.

  7. Vibroacoustic Analysis of a Rectangular Enclosure Bounded by a Flexible Panel with Clamped Boundary Condition

    Directory of Open Access Journals (Sweden)

    Yuan Wang

    2014-01-01

    Full Text Available A lot of research on the properties of a panel-enclosure coupled system which consists of an enclosure with a simply supported flexible wall was conducted. However, an enclosure with clamped flexible walls is commonly encountered. The properties of a panel-enclosure system, with one clamped flexible wall, are different from those of a simply supported one, and there are only a few studies on it. Thus, this paper is dedicated to the study of the effect of structural-acoustic coupling on the properties of a panel-enclosure system, which consists of an enclosure with a clamped panel. The resonance frequencies and the decay times of the coupled system are obtained using the classical modal coupling theory. The effects of enclosure and its flexible boundary on the resonance frequencies and the modal decay times of the coupled system are then analyzed. It is shown that, when panel thickness is changed, coupling strength is determined by the difference between the resonance frequencies of the panel and the enclosure modes. However, for the variation of enclosure depth, the factor which determines the coupling strength between panel and enclosure modes is the enclosure depth or the difference between their resonance frequencies.

  8. Bipolar Thermofusion BiClamp 150 in Thyroidectomy: A Review of 1156 Operations

    Directory of Open Access Journals (Sweden)

    Tomáš Pniak

    2014-01-01

    Full Text Available Objectives. To compare the bipolar thermofusion BiClamp 150 with conventional ligature techniques for thyroid gland surgery, and report the advantages/disadvantages of both techniques. Methods. In this retrospective comparative study, all thyroid gland operations performed in the ENT Clinic Faculty Hospital Ostrava from 2006 to 2013 were included (1156 operations, 2122 lobes. Patients were categorized into two groups according to the type of vessel sealing method used, group I (BiClamp, n=819 operations and group II (conventional ligature, n=337 operations. The number of revision surgeries due to wound hematoma was recorded as a bleeding event. Statistical analysis of the complication rate (bleeding rate, recurrent nerve palsy and time of duration was performed. Results. The rate of revision surgery performed due to postoperative wound hematoma was significantly lower in group I (15/819, 1.83% compared with group II (14/337, 4.15% (P=0.022. There was no statistically significant difference in the frequency of recurrent laryngeal nerve palsy between groups I and II (P=0.36. The average surgery time was significantly shorter in group I (P<0.001. Conclusion. Bipolar thermofusion BiClamp is an effective vessel sealing method that leads to a significant reduction in postoperative wound bleeding rates and reduces surgical time compared with conventional vessel ligature.

  9. Histone locus regulation by the Drosophila dosage compensation adaptor protein CLAMP.

    Science.gov (United States)

    Rieder, Leila E; Koreski, Kaitlin P; Boltz, Kara A; Kuzu, Guray; Urban, Jennifer A; Bowman, Sarah K; Zeidman, Anna; Jordan, William T; Tolstorukov, Michael Y; Marzluff, William F; Duronio, Robert J; Larschan, Erica N

    2017-07-15

    The conserved histone locus body (HLB) assembles prior to zygotic gene activation early during development and concentrates factors into a nuclear domain of coordinated histone gene regulation. Although HLBs form specifically at replication-dependent histone loci, the cis and trans factors that target HLB components to histone genes remained unknown. Here we report that conserved GA repeat cis elements within the bidirectional histone3-histone4 promoter direct HLB formation in Drosophila In addition, the CLAMP (chromatin-linked adaptor for male-specific lethal [MSL] proteins) zinc finger protein binds these GA repeat motifs, increases chromatin accessibility, enhances histone gene transcription, and promotes HLB formation. We demonstrated previously that CLAMP also promotes the formation of another domain of coordinated gene regulation: the dosage-compensated male X chromosome. Therefore, CLAMP binding to GA repeat motifs promotes the formation of two distinct domains of coordinated gene activation located at different places in the genome. © 2017 Rieder et al.; Published by Cold Spring Harbor Laboratory Press.

  10. External mean flow influence on sound transmission through finite clamped double-wall sandwich panels

    Science.gov (United States)

    Liu, Yu; Catalan, Jean-Cédric

    2017-09-01

    This paper studies the influence of an external mean flow on the sound transmission through finite clamped double-wall sandwich panels lined with poroelastic materials. Biot's theory is employed to describe wave propagation in poroelastic materials and various configurations of coupling the poroelastic layer to the facing plates are considered. The clamped boundary of finite panels are dealt with by the modal superposition theory and the weighted residual (Garlekin) method, leading to a matrix equation solution for the sound transmission loss (STL) through the structure. The theoretical model is validated against existing theories of infinite sandwich panels with and without an external flow. The numerical results of a single incident wave show that the external mean flow has significant effects on the STL which are coupled with the clamped boundary effect dominating in the low-frequency range. The external mean flow also influences considerably the limiting incidence angle of the panel system and the effect of the incidence angle on the STL. However, the influences of the azimuthal angle and the external flow orientation are negligible.

  11. Thermal and Efficiency Analysis of Five-Level Multilevel-Clamped Multilevel Converter Considering Grid Codes

    DEFF Research Database (Denmark)

    Ma, Ke; Muñoz-Aguilar, R. S.; Rodriguez, Pedro

    2014-01-01

    In this paper, a five-level multilevel-clamped multilevel converter (5L- MLC2) topology is analyzed based on a typical grid integration application for renewable energy system. The loss and thermal distributions of the power devices in different switching legs are investigated and illustrated und...... the clamping switching legs and the main switching legs; it also shows a good and consistent efficiency characteristic under different output power ranges when complying with the grid codes.......In this paper, a five-level multilevel-clamped multilevel converter (5L- MLC2) topology is analyzed based on a typical grid integration application for renewable energy system. The loss and thermal distributions of the power devices in different switching legs are investigated and illustrated under...... rated condition. Afterward, a loss and efficiency evaluation method is proposed and applied which takes into account various injected reactive power ranges regulated by the grid codes. It is concluded that the loss and thermal distributions of the 5L- MLC2 multilevel topology are unequal between...

  12. Is Near-Infrared Spectroscopy a Reliable Method to Evaluate Clamping Ischemia during Carotid Surgery?

    Directory of Open Access Journals (Sweden)

    Luciano Pedrini

    2012-01-01

    Full Text Available Guidelines do not include cerebral oximetry among monitoring for carotid endarterectomy (CEA. The purpose of this study was to evaluate the reliability of near-infrared spectroscopy (NIRS in the detection of clamping ischemia and in the prevention of clamping-related neurologic deficits using, as a cutoff for shunting, a 20% regional cerebral oxygen saturation (rSO2 decrease if persistent more than 4 minutes, otherwise a 25% rSO2 decrease. Bilateral rSO2 was monitored continuously in patients undergoing CEA under general anesthesia (GA. Data was recorded after clamping, declamping, during shunting and lowest values achieved. Preoperative neurologic, CT-scan, and vascular lesions were recorded. We reviewed 473 cases: 305 males (64.5% mean age 73.3±7.3. Three patients presented transient ischemic deficits at awakening, no perioperative stroke or death; 41 (8.7% required shunting: 30 based on the initial rSO2 value and 11 due to a decrease during surgery. Using the ROC curve analysis we found, for a >25% reduction from baseline value, a sensitivity of 100% and a specificity of 90.6%. Reliability, PPV, and NPV were 95.38%, 9%, and 100%, respectively. In conclusion, this study indicates the potential reliability of NIRS monitoring during CEA under GA, using a cutoff of 25% or a cutoff of 20% for prolonged hypoperfusion.

  13. Look twice before you clamp: decapitation of an omphaloenteric duct. A case report.

    Science.gov (United States)

    van Tuil, Cornelia; Saxena, Amulya K; Willital, Gunter H

    2006-01-01

    To report a case of accidental decapitation of a persistent omphaloenteric duct (POD) in congenital hernia of the umbilical cord in a newborn male. The newborn was transferred to an intensive care unit, where under closer inspection the diagnosis of an omphalocele was presumed. Surgical exploration at our Pediatric Surgical University Clinic revealed a clamped and decapitated POD in the umbilical cord. The decapitation had gone unnoticed due to severe asphyxia resulting from meconium aspiration encountered at the time of delivery. Using general anesthesia, the clamp was removed under sterile conditions. The decapitation of the POD in the herniated umbilical cord was confirmed at this time. The severed intestine was sutured and closed using 5-0 Vicryl sutures. The further course was uneventful. Herniation of the umbilical cord at the time of birth by intestinal loops or POD is rare, but caution should be taken by midwives before clamping in order to avoid the type of injury seen in this case. (c) 2006 S. Karger AG, Basel

  14. The effect of whole grain wheat sourdough bread consumption on serum lipids in healthy normoglycemic/normoinsulinemic and hyperglycemic/hyperinsulinemic adults depends on presence of the APOE E3/E3 genotype: a randomized controlled trial

    Directory of Open Access Journals (Sweden)

    Bakovic Marica

    2010-05-01

    Full Text Available Abstract Background Epidemiological studies associate consumption of whole grain foods, including breads, with reduced cardiovascular disease (CVD risk; however, few studies have compared wheat whole grains with wheat refined grains. Methods This study investigated effects of 6-week consumption of whole grain wheat sourdough bread in comparison to white bread on fasting serum lipids in normoglycemic/normoinsulinemic (NGI; n = 14 and hyperglycemic/hyperinsulinemic (HGI; n = 14 adults. The influence of single-nucleotide polymorphisms, 3 within the APOE gene (E2, E3, E4 and 2 within the hepatic lipase gene promoter (LIPC -514C>T, LIPC -250G>A were considered. Results At baseline, HGI participants had significantly higher body weight, waist circumference, body fat, and fasted glucose, insulin, homeostasis model assessment of insulin resistance (HOMA-IR, glucagon, triacylglycerols (TAG and TAG:HDL-cholesterol, compared to NGI participants; however, none of these in addition to none of the other serum lipids, differed between bread treatments, within either participant group. For participants with the APOE E3/E3 genotype, LDL-cholesterol (P = 0.02 increased in the NGI group (n = 7, and TAG (P = 0.03 and TAG:HDL-cholesterol (P = 0.04 increased in the HGI group (n = 10, following consumption of whole grain wheat sourdough compared to white bread. Conclusions In summary, 6-week consumption of whole grain wheat sourdough bread did not significantly modulate serum lipids in NGI or HGI adults; however, it significantly increased LDL-cholesterol, TAG and TAG:HDL-cholesterol in participants with the APOE E3/E3 genotype. These data add to limited literature comparing wheat whole grains to wheat refined grains on CVD risk and highlight the need to consider genetic variation in relation to lipoprotein lipid content and CVD risk.

  15. Timing of clamping and factors associated with iron stores in full-term newborns

    Science.gov (United States)

    Oliveira, Fabiana de Cássia Carvalho; Assis, Karine Franklin; Martins, Mariana Campos; do Prado, Mara Rúbia Maciel Cardoso; Ribeiro, Andréia Queiroz; Sant’Ana, Luciana Ferreira da Rocha; Priore, Silvia Eloiza; Franceschini, Sylvia do Carmo Castro

    2014-01-01

    OBJECTIVE To analyze the impact of timing of clamping and obstetric, biological and socioeconomic factors on the iron stores of full-term newborns. METHODS Cross-sectional study between October 2011 and July 2012 in which hematological parameters were evaluated for newborns in Viçosa, MG, Southeastern Brazil. It involved collecting 7 mL of umbilical cord blood from 144 full-term not underweight newborns. The parameters investigated were complete blood count, serum iron, ferritin and C-reactive protein. The time of umbilical cord clamping was measured using a digital timer without interfering in the procedures of childbirth. The birth data were collected from Live Birth Certificates and other information was obtained from the mother through a questionnaire applied in the first month postpartum. Analysis of multiple linear regression was then used to estimate the influence of biological, obstetrics and socioeconomic factors on the ferritin levels at birth. RESULTS The median ferritin was 130.3 µg/L (n = 129, minimum = 16.4; maximum = 420.5 µg/L), the mean serum iron was 137.9 μg/dL (n = 144, SD = 39.29) and mean hemoglobin was 14.7 g/dL (n = 144, SD = 1.47). The median time of cord clamping was 36 seconds, ranging between 7 and 100. The bivariate analysis detected an association between ferritin levels and color of the child, timing clamping of 60 seconds, type of delivery, the presence of gestational diabetes and per capita family income. In multivariate analysis, the variables per capita income, number of antenatal visits and length at birth accounted for 22.0% of variation in ferritin levels. CONCLUSIONS Iron stores at birth were influenced by biological, obstetric and social characteristics. Tackling anemia should involve creating policies aimed at reducing social inequalities, improving the quality of antenatal care, as well as implementing a criterion of delayed clamping of the umbilical cord within the guidelines of labor. PMID:24789632

  16. Experimental determination of optimal clamping torque for AB-PEM Fuel cell

    Directory of Open Access Journals (Sweden)

    Noor Ul Hassan

    2016-04-01

    Full Text Available Polymer electrolyte Membrane (PEM fuel cell is an electrochemical device producing electricity by the reaction of hydrogen and oxygen without combustion. PEM fuel cell stack is provided with an appropriate clamping torque to prevent leakage of reactant gases and to minimize the contact resistance between gas diffusion media (GDL and bipolar plates. GDL porous structure and gas permeability is directly affected by the compaction pressure which, consequently, drastically change the fuel cell performance. Various efforts were made to determine the optimal compaction pressure and pressure distributions through simulations and experimentation. Lower compaction pressure results in increase of contact resistance and also chances of leakage. On the other hand, higher compaction pressure decreases the contact resistance but also narrows down the diffusion path for mass transfer from gas channels to the catalyst layers, consequently, lowering cell performance. The optimal cell performance is related to the gasket thickness and compression pressure on GDL. Every stack has a unique assembly pressure due to differences in fuel cell components material and stack design. Therefore, there is still need to determine the optimal torque value for getting the optimal cell performance. This study has been carried out in continuation of deve­lopment of Air breathing PEM fuel cell for small Unmanned Aerial Vehicle (UAV application. Compaction pressure at minimum contact resistance was determined and clamping torque value was calcu­la­ted accordingly. Single cell performance tests were performed at five different clamping torque values i.e 0.5, 1.0, 1.5, 2.0 and 2.5 N m, for achieving optimal cell per­formance. Clamping pressure distribution tests were also performed at these torque values to verify uniform pressure distribution at optimal torque value. Experimental and theoretical results were compared for making inferences about optimal cell perfor­man­ce. A

  17. Evaluation of insulin secretion and action in New World camelids.

    Science.gov (United States)

    Firshman, Anna M; Cebra, Christopher K; Schanbacher, Barbara J; Seaquist, Elizabeth R

    2013-01-01

    To measure and compare insulin secretion and sensitivity in healthy alpacas and llamas via glucose clamping techniques. 8 llamas and 8 alpacas. Hyperinsulinemic euglycemic clamping (HEC) and hyperglycemic clamping (HGC) were performed on each camelid in a crossover design with a minimum 48-hour washout period between clamping procedures. The HEC technique was performed to measure insulin sensitivity. Insulin was infused IV at 6 mU/min/kg for 4 hours, and an IV infusion of glucose was adjusted to maintain blood glucose concentration at 150 mg/dL. Concentrations of blood glucose and plasma insulin were determined throughout. The HGC technique was performed to assess insulin secretion in response to exogenous glucose infusion. An IV infusion of glucose was administered to maintain blood glucose concentration at 320 mg/dL for 3 hours, and concentrations of blood glucose and plasma insulin were determined throughout. Alpacas and llamas were not significantly different with respect to whole-body insulin sensitivity during HEC or in pancreatic β-cell response during HGC. Alpacas and llamas had markedly lower insulin sensitivity during HEC and markedly lower pancreatic β-cell response during HGC, in comparison with many other species. New World camelids had lower glucose-induced insulin secretion and marked insulin resistance in comparison with other species. This likely contributes to the disorders of fat and glucose metabolism that are common to camelids.

  18. High conversion ratio DC-DC converter with isolated transformer and switched-clamp capacitor for Taiwan photon source

    Science.gov (United States)

    Wong, Y.-S.; Chen, J.-F.; Liu, K.-B.; Hsieh, Y.-P.

    2017-12-01

    A new high step-up voltage converter that combines a switch capacitor and isolated transformer, together with a passive clamp circuit, is employed to reduce voltage stress on the main power switch. The voltage stress of the power switch should be clamped to 1/4 Vo, and the proposed converter can achieve high step-up voltage gain with appropriate duty ratio. The energy of the leakage inductor can be recycled by the clamp capacitor because of the passive clamp circuit, and low On-state resistance RDS(on) of the power switch can be adopted to reduce the conduction loss. In this paper, several mathematical derivations are presented, CCM and DCM operating principle are discussed, and experimental results are provided to verify the effectiveness of converter topology. Finally, a 24-V-input voltage to 200-V-output voltage and a 150 W output power prototype converter are fabricated in the laboratory.

  19. Association between oxidative stress and cord serum lipids in relation to delayed cord clamping in term neonates.

    Science.gov (United States)

    Moustafa, Asmaa N; Ibrahim, Mahmoud H; Mousa, Suzan Omar; Hassan, Ebtesam E; Mohamed, Hashem F; Moness, Hend M

    2017-11-09

    Although delayed cord clamping (DCC) is a recent WHO recommendation, early cord clamping (ECC) is still a routine practice in many countries. Limited researches studied the effect of delayed cord clamping on oxidative stress in term neonates; In this study we aim to assess the influence of cord clamping either early or late on oxidative stress in term neonates and to evaluate the association of oxidative stress and cord blood lipids. One-hundred mothers and their term neonates were included in the present study. Umbilical cord blood samples were collected from the umbilical vein and umbilical artery immediately following labor. Total cholesterol, total triglycerides and phospholipids levels were significantly higher in the ECC group than the DCC group (p cord blood lipids and an augmented antioxidant activity. This suggests the protective effect of DCC on the future health of the term neonates and supports the application of DCC in active management of 3rd stage of labor in term neonates.

  20. Structure-function analysis of the C-clamp of TCF/Pangolin in Wnt/ß-catenin signaling.

    Directory of Open Access Journals (Sweden)

    Aditi J Ravindranath

    Full Text Available The evolutionarily conserved Wnt/ß-catenin (Wnt/ß-cat pathway plays an important role in animal development in metazoans. Many Wnt targets are regulated by members of the TCF/LEF1 (TCF family of transcription factors. All TCFs contain a High Mobility Group (HMG domain that bind specific DNA sequences. Invertebrate TCFs and some vertebrate TCF isoforms also contain another domain, called the C-clamp, which allows TCFs to recognize an additional DNA motif known as the Helper site. While the C-clamp has been shown to be important for regulating several Wnt reporter genes in cell culture, its physiological role in regulating Wnt targets is less clear. In addition, little is known about this domain, except that two of the four conserved cysteines are functionally important. Here, we carried out a systematic mutagenesis and functional analysis of the C-clamp from the Drosophila TCF/Pangolin (TCF/Pan protein. We found that the C-clamp is a zinc-binding domain that is sufficient for binding to the Helper site. In addition to this DNA-binding activity, the C-clamp also inhibits the HMG domain from binding its cognate DNA site. Point mutations were identified that specifically affected DNA-binding or reduced the inhibitory effect. These mutants were characterized in TCF/Pan rescue assays. The specific DNA-binding activity of the C-clamp was essential for TCF/Pan function in cell culture and in patterning the embryonic epidermis of Drosophila, demonstrating the importance of this C-clamp activity in regulating Wnt target gene expression. In contrast, the inhibitory mutation had a subtle effect in cell culture and no effect on TCF/Pan activity in embryos. These results provide important information about the functional domains of the C-clamp, and highlight its importance for Wnt/ß-cat signaling in Drosophila.

  1. Structure-function analysis of the C-clamp of TCF/Pangolin in Wnt/ß-catenin signaling.

    Science.gov (United States)

    Ravindranath, Aditi J; Ravindranath, Aditi; Cadigan, Ken M

    2014-01-01

    The evolutionarily conserved Wnt/ß-catenin (Wnt/ß-cat) pathway plays an important role in animal development in metazoans. Many Wnt targets are regulated by members of the TCF/LEF1 (TCF) family of transcription factors. All TCFs contain a High Mobility Group (HMG) domain that bind specific DNA sequences. Invertebrate TCFs and some vertebrate TCF isoforms also contain another domain, called the C-clamp, which allows TCFs to recognize an additional DNA motif known as the Helper site. While the C-clamp has been shown to be important for regulating several Wnt reporter genes in cell culture, its physiological role in regulating Wnt targets is less clear. In addition, little is known about this domain, except that two of the four conserved cysteines are functionally important. Here, we carried out a systematic mutagenesis and functional analysis of the C-clamp from the Drosophila TCF/Pangolin (TCF/Pan) protein. We found that the C-clamp is a zinc-binding domain that is sufficient for binding to the Helper site. In addition to this DNA-binding activity, the C-clamp also inhibits the HMG domain from binding its cognate DNA site. Point mutations were identified that specifically affected DNA-binding or reduced the inhibitory effect. These mutants were characterized in TCF/Pan rescue assays. The specific DNA-binding activity of the C-clamp was essential for TCF/Pan function in cell culture and in patterning the embryonic epidermis of Drosophila, demonstrating the importance of this C-clamp activity in regulating Wnt target gene expression. In contrast, the inhibitory mutation had a subtle effect in cell culture and no effect on TCF/Pan activity in embryos. These results provide important information about the functional domains of the C-clamp, and highlight its importance for Wnt/ß-cat signaling in Drosophila.

  2. The influence of delayed cord clamping and cord milking on inflammatory cytokines in umbilical vein and neonatal circulation.

    Science.gov (United States)

    Daskalakis, George; Papapanagiotou, Angeliki; Siristatidis, Charalambos; Drakakis, Petros; Mole, Ioanna; Barbouni, Anastasia; Papavassiliou, Athanasios G; Pergialiotis, Vasilios

    2018-01-16

    The purpose of the present study was to compare the levels of tumor necrosis factor a (TNF-a), and interleukins (IL) 1, 6, 8 and 10 in the umbilical cord and neonatal circulation among neonates with early and late cord clamping MATERIAL AND METHODS: A consecutive series of 76 cases that had an uncomplicated pregnancy and an uneventful parturition was evaluated. In 40 cases delayed cord clamping was used and in the remaining 36 cases early cord clamping was practiced. Blood samples were collected from the umbilical vein immediately after cord clamping and at 24 hours from the median cubital or basilic vein of the neonate RESULTS: Significant differences were noted in the hematocrit and hemoglobin levels at 24 hours that favored delayed clamping. None of the evaluated markers of inflammation differ between the two groups. Spearman's rho revealed a significant correlation between umbilical cord TNF-a and TNF-a neonatal values at 24 hours (r=.551, p=.022) in the early clamped group. Significant correlations were also noted between umbilical cord TNF-a and TNF-a neonatal values at 24 hours (r=.728, p=.001), umbilical cord IL-10 and neonatal IL-10 at 24 hours (r=.487, p=.047) and umbilical cord IL-1b and neonatal IL-1b at 24 hours (r=.516, p=.034) CONCLUSIONS: Delayed cord clamping or cord milking does not alter the levels of inflammatory cytokines in cord blood and neonatal serum. Future studies should evaluate the impact of delayed cord clamping in selected high-risk pregnancies. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  3. A Mouse Model of Ischemic Spinal Cord Injury with Delayed Paralysis Caused by Aortic Cross-clamping

    Science.gov (United States)

    Awad, Hamdy; Ankeny, Daniel P.; Guan, Zhen; Wei, Ping; McTigue, Dana M.; Popovich, Phillip G.

    2010-01-01

    Background Spinal cord ischemia and paralysis are devastating perioperative complications that can accompany open or endovascular repair surgery for aortic aneurysms. Here, we report on the development of a new mouse model of spinal cord ischemia with delayed paralysis induced by cross-clamping the descending aorta. Methods Transient aortic occlusion was produced in mice by cross clamping the descending aorta through a lateral thoracotomy. To establish an optimal surgical procedure with limited mortality, variable cross-clamp times and core temperatures were tested between experiments. Results The onset of paresis or paralysis and postsurgical mortality varied as a function of cross-clamp time and core temperature that was maintained during the period of cross-clamp. Using optimal surgical parameters (7.5 min cross-clamp duration @ 33°C core temperature), the onset of paralysis is delayed 24–36 h postreperfusion and > 95% of mice survive through 9 weeks postsurgery. These mice are further stratified into two groups, with 70% (n = 19/27) of mice developing severe hindlimb paralysis and the remaining mice showing mild, though still permanent, behavioral deficits. Conclusion This new model should prove useful as a preclinical tool for screening neuroprotective therapeutics and for defining the basic biological mechanisms that cause delayed paralysis and neurodegeneration after transient spinal cord ischemia. PMID:20808212

  4. Outcomes of peripheral perfusion with balloon aortic clamping for totally endoscopic robotic mitral valve repair.

    Science.gov (United States)

    Ward, Alison F; Loulmet, Didier F; Neuburger, Peter J; Grossi, Eugene A

    2014-12-01

    Although the technique of totally endoscopic robotic mitral valve repair (TERMR) has been well described, few reports have examined the results of peripheral perfusion with balloon clamping. We analyzed the outcomes of TERMR performed using this strategy. A total of 108 consecutive patients underwent TERMR by a 2-surgeon team. The preoperative evaluation included chest computed tomography and abdominal and pelvis computed tomography. Additional procedures included appendage exclusion in 96, patent foramen ovale closure in 29, cryoablation in 16, tricuspid valve repair in 2, and septal myectomy in 2. The mean patient age was 59 years (range, 21-86). Central venous drainage was obtained with a long cannula. Arterial return was achieved with femoral cannulation, when possible. An endoballoon catheter was placed through the femoral artery. Transesophageal echocardiography was used to position all catheters. Femoral artery perfusion was possible in 103 of 108 patients (95.3%). The subclavian artery was used in 5 patients (4.6%) with contraindications to retrograde perfusion. An endoballoon clamp was placed by way of the femoral artery. In 105 of 108 patients (97.2%), endoaortic occlusion was successfully used; the mean crossclamp time was 87.4 minutes. The coronary sinus cardioplegia catheter was placed successfully in 81 of the 108 patients (75%). Postoperatively, no or mild inotropic support was needed in 94 (87%) and moderate support in 14 (13.0%). Of the 108 patients, 55 (50.9%) were extubated in the operating room. No hospital mortality, aortic injury, vascular complications, or wound infections occurred. Complications included 2 strokes (no residual deficit) (1.8%) and atrial fibrillation in 18 (16.7%). The median hospital stay was 4 days. Eighty patients (74.1%) were discharged by postoperative day 5. A preoperative image-guided perfusion strategy and aortic balloon clamping permit routine TERMR with excellent myocardial preservation and minimal complications

  5. Identification and Characterization of an Inside-Out Folding Intermediate of T4 Phage Sliding Clamp.

    Science.gov (United States)

    Singh, Manika Indrajit; Jain, Vikas

    2017-10-17

    Protein folding process involves formation of transiently occurring intermediates that are difficult to isolate and characterize. It is both necessary and interesting to characterize the structural conformations adopted by these intermediates, also called molten globules (MG), to understand protein folding. Here, we investigated the equilibrium (un)folding intermediate state of T4 phage gene product 45 (gp45, also known as DNA polymerase processivity factor or sliding clamp) obtained during chemical denaturation. We show that gp45 undergoes substantial conformational rearrangement during unfolding and forms an expanded dry-MG. By monitoring the fluorescence of tryptophans that were strategically introduced at various sites, we demonstrate that the urea-treated molecule has its surface residues flip inside the core, and closely placed residues move farther. We were also able to isolate and purify the MG form of gp45 in native condition (i.e., nondenaturing buffer, at physiological pH and temperature); characteristics of this purified molecule substantially match with urea-treated wild-type gp45. To the best of our knowledge, this is one of the few reports that demonstrate the isolation and purification of a protein folding intermediate in native condition. We believe that our work not only allows us to dissect the process of protein folding, but will also help in the designing of folding inhibitors against sliding clamps to treat a wide variety of diseases from bacterial infection to cancer, due to the vast presence of clamps in all the domains of life. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  6. Transition from fetal to neonatal circulation: Modeling the effect of umbilical cord clamping.

    Science.gov (United States)

    Yigit, Mehmet B; Kowalski, William J; Hutchon, David J R; Pekkan, Kerem

    2015-06-25

    Hemodynamics of the fetal to neonatal transition are orchestrated through complex physiological changes and results in cardiovascular adaptation to the adult biventricular circulation. Clinical practice during this critical period can influence vital organ physiology for normal newborns, premature babies and congenital heart defect patients. Particularly, the timing of the cord clamping procedure, immediate (ICC) vs. delayed cord clamping (DCC), is hypothesized to be an important factor for the transitory fetal hemodynamics. The clinical need for a quantitative understanding of this physiology motivated the development of a lumped parameter model (LPM) of the fetal cardio-respiratory system covering the late-gestation to neonatal period. The LPM was validated with in vivo clinical data and then used to predict the effects of cord clamping procedures on hemodynamics and vital gases. Clinical time-dependent resistance functions to simulate the vascular changes were introduced. For DCC, placental transfusion (31.3 ml) increased neonatal blood volume by 11.7%. This increased blood volume is reflected in an increase in preload pressures by ~20% compared to ICC, which in turn increased the cardiac output (CO) by 20% (COICC=993 ml/min; CODCC=1197 ml/min). Our model accurately predicted dynamic flow patterns in vivo. DCC was shown to maintain oxygenation if the onset of pulmonary respiration was delayed or impaired. On the other hand, a significant 25% decrease in oxygen saturations was observed when applying ICC under the same physiological conditions. We conclude that DCC has a significant impact on newborn hemodynamics, mainly because of the improved blood volume and the sustained placental respiration. Copyright © 2015. Published by Elsevier Ltd.

  7. Near-total pancreatectomy for persistent hyperinsulinemic ...

    African Journals Online (AJOL)

    All of them suffered from symptoms of PHHI: thermoregulatory problems in five cases, tremors in five cases, seizures in three, irritability in five, respiratory distress in eight, apnea in four, hypotonia in four, lethargy in five, and feeding difficulty in four cases. All cases were diagnosed following strict criteria that included bouts of ...

  8. Near-total pancreatectomy for persistent hyperinsulinemic ...

    African Journals Online (AJOL)

    medical treatment. Their ages at the time of surgery ranged from 20 days to 27 months. Only patients who were surgically managed were included in this study. All of them suffered from symptoms of PHHI: thermoregulatory problems in five cases, tremors in five cases, seizures in three, irritability in five, respiratory distress in ...

  9. Transverse Vibrations of Clamped and Simply-Supported Circular Plates with Two Dimensional Thickness Variation

    Directory of Open Access Journals (Sweden)

    N. Bhardwaj

    2012-01-01

    Full Text Available Two dimensional boundary characteristic orthonormal polynomials are used in the Ritz method for the vibration analysis of clamped and simply-supported circular plates of varying thickness. The thickness variation in the radial direction is linear whereas in the circumferential direction the thickness varies according to cos kθ, where k is an integer. In order to verify the validity, convergence and accuracy of the results, comparison studies are made against existing results for the special case of linearly tapered thickness plates. Variations in frequencies for the first six normal modes of vibration and mode shapes for various taper parameters are presented.

  10. Characterization of a patch-clamp microchannel array towards neuronal networks analysis

    DEFF Research Database (Denmark)

    Alberti, Massimo; Snakenborg, Detlef; Lopacinska, Joanna M.

    2010-01-01

    for simultaneous patch clamping of cultured cells or neurons in the same network. A disposable silicon/silicon dioxide (Si/SiO2) chip with a microhole array was integrated in a microfluidic system for cell handling, perfusion and electrical recording. Fluidic characterization showed that our PC mu CA can work...... theory, but the capacitance is still too high for high resolution recording. The system was tested on HeLa cells: successful cell trapping with a sealing of 40 M Omega was recorded. Modification of the Si/SiO2 chip is needed in order to achieve a better sealing and long-term cell culturing in the PC mu...

  11. Direct-conversion switching-mode audio power amplifier with active capacitive voltage clamp

    Energy Technology Data Exchange (ETDEWEB)

    Ljusev, P.; Andersen, Michael A.E.

    2005-07-01

    This paper discusses the advantages and problems when implementing direct energy conversion switching-mode audio power amplifiers. It is shown that the total integration of the power supply and Class D audio power amplifier into one compact direct converter can simplify design, increase efficiency and integration level, reduce product volume and lower its cost. As an example, the principle of operation and the measurements made on a direct-conversion switching-mode audio power amplifier with active capacitive voltage clamp are presented. (au)

  12. Activity Clamp Provides Insights into Paradoxical Effects of the Anti-Seizure Drug Carbamazepine.

    Science.gov (United States)

    Morris, Gareth; Leite, Marco; Kullmann, Dimitri M; Pavlov, Ivan; Schorge, Stephanie; Lignani, Gabriele

    2017-05-31

    A major challenge in experimental epilepsy research is to reconcile the effects of anti-epileptic drugs (AEDs) on individual neurons with their network-level actions. Highlighting this difficulty, it is unclear why carbamazepine (CBZ), a frontline AED with a known molecular mechanism, has been reported to increase epileptiform activity in several clinical and experimental studies. We confirmed in an in vitro mouse model (in both sexes) that the frequency of interictal bursts increased after CBZ perfusion. To address the underlying mechanisms, we developed a method, activity clamp, to distinguish the response of individual neurons from network-level actions of CBZ. We first recorded barrages of synaptic conductances from neurons during epileptiform activity and then replayed them in pharmacologically isolated neurons under control conditions and in the presence of CBZ. CBZ consistently decreased the reliability of the second action potential in each burst of activity. Conventional current-clamp recordings using excitatory ramp or square-step current injections failed to reveal this effect. Network modeling showed that a CBZ-induced decrease of neuron recruitment during epileptic bursts can lead to an increase in burst frequency at the network level by reducing the refractoriness of excitatory transmission. By combining activity clamp with computer simulations, the present study provides a potential explanation for the paradoxical effects of CBZ on epileptiform activity.SIGNIFICANCE STATEMENT The effects of anti-epileptic drugs on individual neurons are difficult to separate from their network-level actions. Although carbamazepine (CBZ) has a known anti-epileptic mechanism, paradoxically, it has also been reported to increase epileptiform activity in clinical and experimental studies. To investigate this paradox during realistic neuronal epileptiform activity, we developed a method, activity clamp, to distinguish the effects of CBZ on individual neurons from network

  13. Off-Clamp Robot-Assisted Partial Nephrectomy: How Far Shall We Proceed?

    Science.gov (United States)

    Abdel Raheem, Ali; Santok, Glen Denmer; Kim, Lawrence H C; Chang, Ki Don; Lum, Trenton G H; Yoon, Young Eun; Han, Woong Kyu; Choi, Young Deuk; Rha, Koon Ho

    2017-10-19

    Off-clamp robot-assisted partial nephrectomy (RAPN) is associated with increased intraoperative blood loss compared with on-clamp technique. Our aim was to demonstrate our surgical technique and to determine which tumors are ideally suited for this technique. Sixty-two patients who underwent off-clamp RAPN for renal tumor between 2006 and 2016 were retrospectively analyzed. Increased estimated blood loss (EBL) volume was defined as more than 75 percentile. receiver operating characteristic (ROC) analysis was used to determine exact cut-off tumor size and the preoperative aspects and dimensions used for an anatomical (PADUA) score that are associated with increased EBL. Risk factors for increased EBL >400 mL and chronic kidney disease (CKD) upstaging were evaluated using logistic regression analysis. The median follow-up period was 20 months (interquartile range [IQR]: 12-84). Patient's mean age, mean tumor size, and mean body mass index were 53.5 ± 12.2 years, 2.6 ± 1.5 cm, and 25 ± 4.1 kg/m2, respectively. Median EBL volume was 200 mL (IQR: 100-400). ROC analysis showed that tumor size of 3.2 cm (area under the curve [AUC] = 0.82, P 400 mL. Patients with tumor size >3.2 cm had longer operative time (116 versus 163 minutes, P = .002), more EBL (150 versus 575 mL, P 400 mL was the only predictor of CKD upstaging (odds ratio: 6.704, P = .009). Our study showed that the risk of intraoperative bleeding and transfusion rate during off-clamp RAPN is increased if tumor size >3.2 cm and/or PADUA complexity score ≥9. Moreover, EBL >400 mL was a risk factor of CKD upstaging, despite zero ischemia. Further larger prospective studies are warranted to validate our results.

  14. Crystal structure of the DNA polymerase III β subunit (β-clamp) from the extremophile Deinococcus radiodurans.

    Science.gov (United States)

    Niiranen, Laila; Lian, Kjersti; Johnson, Kenneth A; Moe, Elin

    2015-02-27

    Deinococcus radiodurans is an extremely radiation and desiccation resistant bacterium which can tolerate radiation doses up to 5,000 Grays without losing viability. We are studying the role of DNA repair and replication proteins for this unusual phenotype by a structural biology approach. The DNA polymerase III β subunit (β-clamp) acts as a sliding clamp on DNA, promoting the binding and processivity of many DNA-acting proteins, and here we report the crystal structure of D. radiodurans β-clamp (Drβ-clamp) at 2.0 Å resolution. The sequence verification process revealed that at the time of the study the gene encoding Drβ-clamp was wrongly annotated in the genome database, encoding a protein of 393 instead of 362 amino acids. The short protein was successfully expressed, purified and used for crystallisation purposes in complex with Cy5-labeled DNA. The structure, which was obtained from blue crystals, shows a typical ring-shaped bacterial β-clamp formed of two monomers, each with three domains of identical topology, but with no visible DNA in electron density. A visualisation of the electrostatic surface potential reveals a highly negatively charged outer surface while the inner surface and the dimer forming interface have a more even charge distribution. The structure of Drβ-clamp was determined to 2.0 Å resolution and shows an evenly distributed electrostatic surface charge on the DNA interacting side. We hypothesise that this charge distribution may facilitate efficient movement on encircled DNA and help ensure efficient DNA metabolism in D. radiodurans upon exposure to high doses of ionizing irradiation or desiccation.

  15. A dnaN Plasmid Shuffle Strain for Rapid In Vivo Analysis of Mutant Escherichia coli β Clamps Provides Insight Into the Role of Clamp in umuDC-Mediated Cold Sensitivity

    Science.gov (United States)

    Babu, Vignesh M. P.; Sutton, Mark D.

    2014-01-01

    The E. coli umuDC gene products participate in two temporally distinct roles: UmuD2C acts in a DNA damage checkpoint control, while UmuD'2C, also known as DNA polymerase V (Pol V), catalyzes replication past DNA lesions via a process termed translesion DNA synthesis. These different roles of the umuDC gene products are managed in part by the dnaN-encoded β sliding clamp protein. Co-overexpression of the β clamp and Pol V severely blocked E. coli growth at 30°C. We previously used a genetic assay that was independent of the ability of β clamp to support E. coli viability to isolate 8 mutant clamp proteins (βQ61K, βS107L, βD150N, βG157S, βV170M, βE202K, βM204K and βP363S) that failed to block growth at 30°C when co-overexpressed with Pol V. It was unknown whether these mutant clamps were capable of supporting E. coli viability and normal umuDC functions in vivo. The goals of this study were to answer these questions. To this end, we developed a novel dnaN plasmid shuffle assay. Using this assay, βD150N and βP363S were unable to support E. coli viability. The remaining 6 mutant clamps, each of which supported viability, were indistinguishable from β+ with respect to umuDC functions in vivo. In light of these findings, we analyzed phenotypes of strains overexpressing either β clamp or Pol V alone. The strain overexpressing β+, but not those expressing mutant β clamps, displayed slowed growth irrespective of the incubation temperature. Moreover, growth of the Pol V-expressing strain was modestly slowed at 30°, but not 42°C. Taken together, these results suggest the mutant clamps were identified due to their inability to slow growth rather than an inability to interact with Pol V. They further suggest that cold sensitivity is due, at least in part, to the combination of their individual effects on growth at 30°C. PMID:24896652

  16. Predictors of cross-clamp-induced intraoperative monitoring changes during carotid endarterectomy using both electroencephalography and somatosensory evoked potentials.

    Science.gov (United States)

    Domenick Sridharan, Natalie; Thirumala, Partha; Chaer, Rabih; Balzer, Jeffrey; Long, Becky; Crammond, Donald; Makaroun, Michel; Avgerinos, Efthymios

    2017-07-05

    The efficacy of selective shunting during carotid endarterectomy (CEA) using intraoperative monitoring (IOM) for detection of cerebral ischemia is well established. There is mounting evidence that monitoring of both electroencephalography (EEG) and somatosensory evoked potentials (SSEPs) increases the sensitivity of cerebral ischemia detection. Predictors of cerebral ischemia requiring selective shunt placement using IOM of both EEG and SSEPs have not been previously identified. Consecutive patients who underwent CEA between January 1, 2000, and December 31, 2010, were retrospectively analyzed. Primary end points were IOM changes at any time during the operation or IOM changes with carotid cross-clamping. Risk factors assessed included demographics; baseline comorbidities; severity of ipsilateral and contralateral disease; symptomatic status; and use of statin, antiplatelet, and beta-blocker medications. Univariate and multivariate logistic regression was used for analysis. During the 11-year study period, a total of 758 patients underwent 804 CEAs (mean age, 70.6 ± 9.5 years; 59.8% male; 39.2% symptomatic) using IOM of both SSEPs and EEG for selective shunting guidance. Postoperative stroke rate was 1.37%; 27.1% of patients had significant SSEP or EEG changes, and 49.1% of these were clamp induced (within 5 minutes of cross-clamping). Of these patients, 83.2% received a shunt (11.4% overall). The most common reason that a shunt was not placed after cross-clamp-induced changes was that the changes resolved with further blood pressure elevation (8 of 17 patients). Clamp-induced IOM changes were predictive of postoperative stroke (odds ratio [OR], 5.5; P = .005). Risk factors for clamp-induced IOM changes were contralateral carotid occlusion (OR, 2.5; P = .01), symptomatic stenosis (OR, 1.8; P = .006), and diabetes (OR, 1.6; P = .03), whereas there was a trend toward increased risk with female sex (OR, 1.5; P = .08). Risk factors for any IOM change (clamp

  17. Shape of a clamped stiff harpsichord wire driven at a resonant frequency

    Science.gov (United States)

    Hanson, Roger J.; Macomber, Hilliard Kent; Boucher, Mathew A.

    2002-05-01

    A wire transversely driven by a sinusoidal force at the resonant frequency of a vibrational mode vibrates at the driving frequency and at harmonics generated by nonlinear processes in the wire. If the amplitude of a harmonic is measured as a function of position along the wire, its shape is revealed. It differs significantly from a sinusoid in the vicinity of either end of the wire because the ends are clamped and the wire has significant stiffness. The shapes of various harmonics have been determined for a brass harpsichord wire, 70 cm long, from optical detector measurements made at different distances from a clamped end. Knowledge of shape facilitates the determination of antinode amplitudes of harmonics when the gross motion of the wire is so large that the detectors must be positioned near an end of the wire because of their very limited dynamic range. Some observations of harmonics and related phenomena were reported previously [Hanson et al., J. Acoust Soc. Am. 108, 2592 (2000); 106, 2141 (1999)]. The shape information is also needed to help separate nonlinear effects possibly occurring in the detectors from those of interest, occurring in the wire itself.

  18. Testing of Diode-Clamping in an Inductive Pulsed Plasma Thruster Circuit

    Science.gov (United States)

    Toftul, Alexandra; Polzin, Kurt A.; Martin, Adam K.; Hudgins, Jerry L.

    2014-01-01

    Testing of a 5.5 kV silicon (Si) diode and 5.8 kV prototype silicon carbide (SiC) diode in an inductive pulsed plasma thruster (IPPT) circuit was performed to obtain a comparison of the resulting circuit recapture efficiency,eta(sub r), defined as the percentage of the initial charge energy remaining on the capacitor bank after the diode interrupts the current. The diode was placed in a pulsed circuit in series with a silicon controlled rectifier (SCR) switch, and the voltages across different components and current waveforms were collected over a range of capacitor charge voltages. Reverse recovery parameters, including turn-off time and peak reverse recovery current, were measured and capacitor voltage waveforms were used to determine the recapture efficiency for each case. The Si fast recovery diode in the circuit was shown to yield a recapture efficiency of up to 20% for the conditions tested, while the SiC diode further increased recapture efficiency to nearly 30%. The data presented show that fast recovery diodes operate on a timescale that permits them to clamp the discharge quickly after the first half cycle, supporting the idea that diode-clamping in IPPT circuit reduces energy dissipation that occurs after the first half cycle

  19. High Performance ZVT with Bus Clamping Modulation Technique for Single Phase Full Bridge Inverters

    Energy Technology Data Exchange (ETDEWEB)

    Xia, Yinglai; Ayyanar, Raja

    2016-03-20

    This paper proposes a topology based on bus clamping modulation and zero-voltage-transition (ZVT) technique to realize zero-voltage-switching (ZVS) for all the main switches of the full bridge inverters, and inherent ZVS and/or ZCS for the auxiliary switches. The advantages of the strategy include significant reduction in the turn-on loss of the ZVT auxiliary switches which typically account for a major part of the total loss in other ZVT circuits, and reduction in the voltage ratings of auxiliary switches. The modulation scheme and the commutation stages are analyzed in detail. Finally, a 1kW, 500 kHz switching frequency inverter of the proposed topology using SiC MOSFETs has been built to validate the theoretical analysis. The ZVT with bus clamping modulation technique of fixed timing and adaptive timing schemes are implemented in DSP TMS320F28335 resulting in full ZVS for the main switches in the full bridge inverter. The proposed scheme can save up to 33 % of the switching loss compared with no ZVT case.

  20. Robotic partial nephrectomy with cold ischemia and on-clamp tumor extraction: recapitulating the open approach.

    Science.gov (United States)

    Rogers, Craig G; Ghani, Khurshid R; Kumar, Ramesh K; Jeong, Wooju; Menon, Mani

    2013-03-01

    We describe a reproducible technique for achieving cold ischemia with intraoperative tumor assessment during robotic partial nephrectomy (RPN) that recapitulates the open approach: intracorporeal cooling and extraction (ICE). A total of seven patients underwent the ICE modification of RPN by transperitoneal (n=5) and retroperitoneal (n=2) approaches. A Gelpoint access port was used for the camera and assistant ports. Following hilar clamping, ice slush was introduced through the Gelpoint via syringes and applied over the kidney surface. The excised tumor was immediately extracted through the Gelpoint, allowing gross margin assessment by pathology during the renorrhaphy. RPN was achieved in all cases with successful introduction of ice slush and tumor extraction while on clamp. Median RENAL nephrometry score was 8 (range: 6-10), and there was one solitary kidney. Mean cold ischemia time was 19.6 min (range: 8-37) and mean estimated blood loss was 296.4 ml (range: 50-1000). Renal parenchymal temperatures cold ischemia and there was no drop in core body temperature >0.5°C during any procedures. Intraoperative assessment of the excised tumor showed adequate gross margins in all cases and final pathology confirmed negative surgical margins. Copyright © 2012 European Association of Urology. Published by Elsevier B.V. All rights reserved.

  1. Vibroacoustic behavior of clamp mounted double-panel partition with enclosure air cavity.

    Science.gov (United States)

    Xin, F X; Lu, T J; Chen, C Q

    2008-12-01

    A theoretical study on the vibroacoustic performance of a rectangular double-panel partition clamp mounted in an infinite acoustic rigid baffle is presented. With the clamped boundary condition taken into account by the method of modal function, a double Fourier series solution to the dynamic response of the structure is obtained by employing the weighted residual method (i.e., the Galerkin method). The double series solution can be considered as the exact solution of the problem, as the structural and acoustic-structural coupling effects are fully accounted for and the solution converges numerically. The accuracy of the theoretical predictions is checked against existing experimental data, with good agreement achieved. The influence of several key parameters on the sound isolation capability of the double-panel configuration is then systematically studied, including panel dimensions, thickness of air cavity, elevation angle, and azimuth angle of incidence sound. The present method is suitable for double-panel systems of finite or infinite extent and is applicable for both low- and high-frequency ranges. With these merits, the proposed method compares favorably with a number of other approaches, e.g., finite element method, boundary element method, and statistical energy analysis method.

  2. Design and control of a novel gastroscope intervention mechanism with circumferentially pneumatic-driven clamping function.

    Science.gov (United States)

    Li, Yanmin; Liu, Hao; Hao, Siwen; Li, Hongyi; Han, Jianda; Yang, Yunsheng

    2017-03-01

    Robot-assisted manipulation is promising for solving problems such as understaffing and the risk of infection in gastro-intestinal endoscopy. However, the commonly used friction rollers in few existing systems have a potential risk of deforming flexible endoscopes for non-uniform clamping. This paper presents a robotic system for a standard flexible endoscope and focuses on a novel gastroscope intervention mechanism (GIM), which provides circumferentially uniform clamping with an airbag. The GIM works with a relay-on mechanism in a way similar to manual operation. The shear stiffness of airbag and the critical slipping force (CSF) were analysed to determine the parameters of the airbag. A fuzzy PID controller was employed to realize a fast response and high accuracy of pneumatic actuation. Experiments were performed to evaluate the accuracy, stiffness and CSF. In vitro and in vivo animal experiments were also carried out. The GIM realized an accuracy of 0.025 ± 0.2 mm and -0.03 ± 0.25° for push-pull and rotation without delivery resistance. Under controlled to guarantee operating safety. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  3. Influence of squeeze film damping on the higher-order modes of clamped–clamped microbeams

    KAUST Repository

    Alcheikh, Nouha

    2016-05-06

    This paper presents an experimental study and a finite-element analysis of the effect of squeeze film damping on the resonance frequency and quality factor of the higher-order flexure vibrations modes of clamped-clamped microbeams. Viscoelastic and silicon nitride microbeams are fabricated and are electrostatically actuated by various electrode configurations to trigger the first, second, and third modes. The damping characteristic and the resonance frequency of these modes are examined for a wide range of gas pressure and electrostatic voltage loads. The results of the silicon nitride beams and viscoelastic beams are compared. It is found that the intrinsic material loss is the major dissipation mechanism at low pressure for the viscoelastic microbeams, significantly limiting their quality factor. It is also found that while the silicon nitride beams show higher quality factors at the intrinsic and molecular regimes of pressure, due to their low intrinsic loss, their quality factors near atmospheric pressure are lower than those of the viscoelastic microbeams. Further, the higher-order modes of all the beams show much higher quality factors at atmospheric pressure compared to the first mode, which could be promising for operating such resonators in air. Experimental results and finite element model simulations show good agreement for resonance frequency and quality factor for the three studied modes. © 2016 IOP Publishing Ltd.

  4. Higher order modes excitation of electrostatically actuated clamped–clamped microbeams: experimental and analytical investigation

    KAUST Repository

    Jaber, Nizar

    2016-01-06

    © 2016 IOP Publishing Ltd. In this study, we demonstrate analytically and experimentally the excitations of the higher order modes of vibrations in electrostatically actuated clamped-clamped microbeam resonators. The concept is based on using partial electrodes with shapes that induce strong excitation of the mode of interest. The devices are fabricated using polyimide as a structural layer coated with nickel from the top and chrome and gold layers from the bottom. Experimentally, frequency sweeps with different electro-dynamical loading conditions are shown to demonstrate the excitation of the higher order modes of vibration. Using a half electrode, the second mode is excited with high amplitude of vibration compared with almost zero response using the full electrode. Also, using a two-third electrode configuration is shown to amplify the third mode resonance amplitude compared with the full electrode under the same electrical loading conditions. An analytical model is developed based on the Euler-Bernollui beam model and the Galerkin method to simulate the device response. Good agreement between the simulation results and the experimental data is reported.

  5. Blood pressure monitoring during exercise: comparison of pulse transit time and volume clamp methods.

    Science.gov (United States)

    Wibmer, Thomas; Denner, Coy; Fischer, Christoph; Schildge, Benedikt; Rüdiger, Stefan; Kropf-Sanchen, Cornelia; Rottbauer, Wolfgang; Schumann, Christian

    2015-01-01

    During physical exercise, pulse transit time (PTT), expressed as the interval between ventricular electrical activity and peripheral pulse wave, may provide a surrogate estimate for blood pressure by the use of specific calibration procedures. The objective of this study was to determine systolic blood pressure (SBP) values derived from the PTT method and from an established method of non-invasive continuous blood pressure measurement based on the volume clamp technique, and to compare their agreement with sphygmomanometry during exercise tests. In 18 subjects, electrocardiogram (ECG) and finger-photoplethysmography were continuously recorded during maximal cycle exercise tests. Intermittent and continuous blood pressure measurements were simultaneously taken using automated sphygmomanometry and a Portapres Model-2 device, respectively. PTT was calculated for each ECG R-wave and the corresponding steepest upstroke slope in the photoplethysmogram, and was transformed to a continuous blood pressure estimate using multipoint nonlinear regression calibration based on the individual subject's sphygmomanometer readings. Bland-Altman limits of agreement between PTT-derived SBP estimates and sphygmomanometer values were -24.7 to 24.1 mmHg, and between Portapres and sphygmomanometer SBP values were -42.0 to 70.1 mmHg. For beat-to-beat SBP estimation during exercise, PTT measurement combined with multipoint nonlinear regression calibration based on intermittent sphygmomanometry may be an alternative to volume clamp devices.

  6. Mechanically clamped PZT ceramics investigated by First-order reversal curves diagram

    Directory of Open Access Journals (Sweden)

    Laurentiu Stoleriu

    2010-09-01

    Full Text Available The First Order Reversal Curves (FORC diagrams method was developed for characterizing the switching properties of ferroelectrics. In the present paper, the FORC method was applied for hard Pb(Zr,TiO3 ceramics with symmetric and asymmetric clamping. An ideal high-oriented single-crystalline ferroelectric with rectangular P(E loop would be characterised by a delta-function FORC distribution, while real ferroelectrics and mostly the polycrystalline ceramics show dispersed FORC distributions. All the investigated ceramics show FORC distributions with non-Gaussian shape, slightly elongated along the coercitive axis, meaning a high dispersion of the energy barriers separating the two bi-stable polarizations ±P. The degree of dispersion is enhanced by clamping. The maximum FORC coercivity is located at ~ (1.9-2 MV/m for all the hard ceramics. The FORC cycling experiment causes the reversal of the initial poling and result in a positive/negative bias on the FORC diagrams. According to the observed features, it results that FORC coercivity is more related to the nature of the material, while the bias field is more sensitive to the electrical and mechanical boundary conditions in which the ferroelectric ceramics evolves while switching.

  7. Experimental model of mesenteric ischemia: reperfusion by abdominal aorta clamping in Wistar rats.

    Science.gov (United States)

    Rocha, Bruno da Costa; Mendes, Rogério Rafael da Silva; Lima, Gabriel Varjão; Albuquerque, Gabriel de Souza; Araújo, Lucas Lacerda; de Jesus, Mateus Neves da Silva; Dos Santos, Washington Luís Conrado; Carreiro, Mário Castro

    2012-01-01

    To develop an experimental model of global normothermic ischemia able to demonstrate the transient ischemia and reperfusion periods required for development of ischemia/reperfusion injury in the small intestines of Wistar rats by clamping the abdominal aorta. Twenty adult male Wistar rats weighing 250-350g were randomly divided into five groups with four rats each and submitted to increasing times of ischemia (0 - 30 - 45 - 60 - 90 minutes). Within each group, except the control one, two rats underwent 60 minutes of reperfusion and two 90 minutes. After the procedures, histological analysis was conducted by measurement of areas of necrosis. The degree of intestinal necrosis ranged from 15% to 54% (p = 0.0004). There was progressive increase in the degree of injury related to increase in ischemic time. However, greater degrees of injury were observed in the lowest times of reperfusion. The analysis of the coefficient of variation of necrosis among the ten groups of ischemia/reperfusion showed a statistically significant difference in 15 areas, 13 related to the control group. The model was able to show the periods required for the occurrence of ischemia/reperfusion injury by aortic clamping and can serve as a basis to facilitate the development of studies that aim at understanding this kind of injury.

  8. A low-voltage sense amplifier with two-stage operational amplifier clamping for flash memory

    Science.gov (United States)

    Guo, Jiarong

    2017-04-01

    A low-voltage sense amplifier with reference current generator utilizing two-stage operational amplifier clamp structure for flash memory is presented in this paper, capable of operating with minimum supply voltage at 1 V. A new reference current generation circuit composed of a reference cell and a two-stage operational amplifier clamping the drain pole of the reference cell is used to generate the reference current, which avoids the threshold limitation caused by current mirror transistor in the traditional sense amplifier. A novel reference voltage generation circuit using dummy bit-line structure without pull-down current is also adopted, which not only improves the sense window enhancing read precision but also saves power consumption. The sense amplifier was implemented in a flash realized in 90 nm flash technology. Experimental results show the access time is 14.7 ns with power supply of 1.2 V and slow corner at 125 °C. Project supported by the National Natural Science Fundation of China (No. 61376028).

  9. Voltage clamp studies on S-layer-supported tetraether lipid membranes.

    Science.gov (United States)

    Schuster, B; Pum, D; Sleytr, U B

    1998-02-02

    Isolated subunits from the cell surface proteins (S-layer) of Bacillus coagulans E38-66 have been recrystallized on a glycerol dialkyl nonitol tetraether lipid (GDNT)-monolayer and the electrophysical features of this biomimetic membrane have been investigated in comparison to unsupported GDNT-monolayers. The GDNT-monolayer, spread on a Langmuir-Blodgett trough, was clamped with the tip of a glass patch pipette. In order to investigate the barrier function and potential to incorporate functional molecules, voltage-clamp examinations on plain and S-layer-supported GDNT-monolayers were per-formed. Our results indicate the formation of a tight GDNT-monolayer sealing the tip of the glass pipette, and a decrease in conductance of the GDNT-monolayer upon recrystallization of the S-layer protein. Thus, the S-layer protein, apparently, did not penetrate or rupture the lipid monolayer. The valinomycin-mediated increase in conductance was less pronounced for the S-layer-supported than for the plain GDNT-monolayer, indicating differences in the accessibility and/or in the fluidity of the lipid membranes. Furthermore. in contrast to plain GDNT-monolayers. S-layer supported GDNT-monolayers with high valinomycin-mediated conductance persisted over long, periods of time, indicating enhanced stability. These composite S-layer/lipid films may constitute a new tool for electrophysical and electrophysiological studies on membrane-associated and membrane-integrated biomolecules.

  10. A Clamped Be Window for the Dump of the HiRadMat Experiment at CERN

    CERN Document Server

    Delonca, M; Grenier, D; Maglioni, C; Sarrio Martinez, A

    2012-01-01

    At CERN, the High Radiation to Materials facility (HiRadMat) is designed to test accelerator components under the impact of high-intensity pulsed beams and will start operation in 2012. In this frame an LHC TED-type dump was installed at the end of the line, working in nitrogen overpressure, and a 254μm-thick beryllium window was placed as barrier between the inside of the dump and the external atmosphere. Because of the special loading conditions, a clamped window design was especially developed, optimized and implemented, the more standard welded window not being suitable for such loads. Considering then the clamping force and the applied differential pressures, the stresses on the window components were carefully evaluated thanks to empirical as well as numerical models, to guarantee the structural integrity of the beryllium foil. This paper reports on choices and optimizations that led to the final design, presenting also comparative results from different solutions and the detailed results for the adopt...

  11. A Clamped Be Window for the Dump of the HiRadMat Experiment at CERN

    CERN Document Server

    Delonca, M; Grenier, D; Maglioni, C; Sarrio Martinez, A

    2012-01-01

    At CERN, the High Radiation to Materials facility (HiRadMat) is designed to test accelerator components under the impact of high-intensity pulsed beams and will start operation in 2012. In this frame an LHC TED-type dump was installed at the end of the line, working in nitrogen over-pressure, and a 254μm-thick beryllium window was placed as barrier between the inside of the dump and the external atmosphere. Because of the special loading conditions, a clamped window design was especially developed, optimized and implemented, the more standard welded window not being suitable for such loads. Considering then the clamping force and the applied differential pressures, the stresses on the window components were carefully evaluated thanks to empirical as well as numerical models, to guarantee the structural integrity of the beryllium foil. This paper reports on choices and optimizations that led to the final design, presenting also comparative results from different solutions and the detailed results for the adop...

  12. The use of clamping grips and friction pads by tree frogs for climbing curved surfaces.

    Science.gov (United States)

    Endlein, Thomas; Ji, Aihong; Yuan, Shanshan; Hill, Iain; Wang, Huan; Barnes, W Jon P; Dai, Zhendong; Sitti, Metin

    2017-02-22

    Most studies on the adhesive mechanisms of climbing animals have addressed attachment against flat surfaces, yet many animals can climb highly curved surfaces, like twigs and small branches. Here we investigated whether tree frogs use a clamping grip by recording the ground reaction forces on a cylindrical object with either a smooth or anti-adhesive, rough surface. Furthermore, we measured the contact area of fore and hindlimbs against differently sized transparent cylinders and the forces of individual pads and subarticular tubercles in restrained animals. Our study revealed that frogs use friction and normal forces of roughly a similar magnitude for holding on to cylindrical objects. When challenged with climbing a non-adhesive surface, the compressive forces between opposite legs nearly doubled, indicating a stronger clamping grip. In contrast to climbing flat surfaces, frogs increased the contact area on all limbs by engaging not just adhesive pads but also subarticular tubercles on curved surfaces. Our force measurements showed that tubercles can withstand larger shear stresses than pads. SEM images of tubercles revealed a similar structure to that of toe pads including the presence of nanopillars, though channels surrounding epithelial cells were less pronounced. The tubercles' smaller size, proximal location on the toes and shallow cells make them probably less prone to buckling and thus ideal for gripping curved surfaces. © 2017 The Authors.

  13. A direct proofreader-clamp interaction stabilizes the Pol III replicase in the polymerization mode

    KAUST Repository

    Jergic, Slobodan

    2013-02-22

    Processive DNA synthesis by the αÉ"θ core of the Escherichia coli Pol III replicase requires it to be bound to the β 2 clamp via a site in the α polymerase subunit. How the É" proofreading exonuclease subunit influences DNA synthesis by α was not previously understood. In this work, bulk assays of DNA replication were used to uncover a non-proofreading activity of É". Combination of mutagenesis with biophysical studies and single-molecule leading-strand replication assays traced this activity to a novel β-binding site in É" that, in conjunction with the site in α, maintains a closed state of the αÉ"θ-β 2 replicase in the polymerization mode of DNA synthesis. The É"-β interaction, selected during evolution to be weak and thus suited for transient disruption to enable access of alternate polymerases and other clamp binding proteins, therefore makes an important contribution to the network of protein-protein interactions that finely tune stability of the replicase on the DNA template in its various conformational states. © 2013 European Molecular Biology Organization.

  14. Voltage clamp methods for the study of membrane currents and SR Ca2+ release in adult skeletal muscle fibres

    Science.gov (United States)

    Hernández-Ochoa, Erick O.; Schneider, Martin F.

    2012-01-01

    Skeletal muscle excitation-contraction (E-C)1 coupling is a process composed of multiple sequential stages, by which an action potential triggers sarcoplasmic reticulum (SR)2 Ca2+ release and subsequent contractile activation. The various steps in the E-C coupling process in skeletal muscle can be studied using different techniques. The simultaneous recordings of sarcolemmal electrical signals and the accompanying elevation in myoplasmic Ca2+, due to depolarization-initiated SR Ca2+ release in skeletal muscle fibres, have been useful to obtain a better understanding of muscle function. In studying the origin and mechanism of voltage dependency of E-C coupling a variety of different techniques have been used to control the voltage in adult skeletal fibres. Pioneering work in muscles isolated from amphibians or crustaceans used microelectrodes or ‘high resistance gap’ techniques to manipulate the voltage in the muscle fibres. The development of the patch clamp technique and its variant, the whole-cell clamp configuration that facilitates the manipulation of the intracellular environment, allowed the use of the voltage clamp techniques in different cell types, including skeletal muscle fibres. The aim of this article is to present an historical perspective of the voltage clamp methods used to study skeletal muscle E-C coupling as well as to describe the current status of using the whole-cell patch clamp technique in studies in which the electrical and Ca2+ signalling properties of mouse skeletal muscle membranes are being investigated. PMID:22306655

  15. An accurate online calibration system based on combined clamp-shape coil for high voltage electronic current transformers.

    Science.gov (United States)

    Li, Zhen-hua; Li, Hong-bin; Zhang, Zhi

    2013-07-01

    Electronic transformers are widely used in power systems because of their wide bandwidth and good transient performance. However, as an emerging technology, the failure rate of electronic transformers is higher than that of traditional transformers. As a result, the calibration period needs to be shortened. Traditional calibration methods require the power of transmission line be cut off, which results in complicated operation and power off loss. This paper proposes an online calibration system which can calibrate electronic current transformers without power off. In this work, the high accuracy standard current transformer and online operation method are the key techniques. Based on the clamp-shape iron-core coil and clamp-shape air-core coil, a combined clamp-shape coil is designed as the standard current transformer. By analyzing the output characteristics of the two coils, the combined clamp-shape coil can achieve verification of the accuracy. So the accuracy of the online calibration system can be guaranteed. Moreover, by employing the earth potential working method and using two insulating rods to connect the combined clamp-shape coil to the high voltage bus, the operation becomes simple and safe. Tests in China National Center for High Voltage Measurement and field experiments show that the proposed system has a high accuracy of up to 0.05 class.

  16. Multiple adjustable vascular clamp prototype: feasibility study on an experimental model of end-to-side microsurgical vascular anastomosis.

    Science.gov (United States)

    Pereira, A; Ichihara, S; Collon, S; Bodin, F; Gay, A; Facca, S; Liverneaux, P

    2014-12-01

    The aim of this study was to establish the feasibility of microsurgical end-to-side vascular anastomosis with a multiclamp adjustable vascular clamp prototype in an inert experimental model. Our method consisted of performing an end-to-side microsurgical anastomosis with 10/0 suture on a 2-mm diameter segment. In group 1, the end-to-side segment was held in place by a double clamp and a single end clamp. In group 2, the segment was held in place with a single multiclamp adjustable clamp. The average time for performing the anastomosis was shorter in group 2. The average number of sutures was the same in both groups. No leak was found and permeability was always positive in both groups. Our results show that performing end-to-side anastomosis with a multiclamp adjustable vascular clamp is feasible in an inert experimental model. Feasibility in a live animal model has to be demonstrated before clinical use. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  17. Nucleic acid clamp-mediated recognition and stabilization of the physiologically relevant MYC promoter G-quadruplex

    Science.gov (United States)

    Hao, Taisen; Gaerig, Vanessa C.; Brooks, Tracy A.

    2016-01-01

    The MYC proto-oncogene is upregulated, often at the transcriptional level, in ∼80% of all cancers. MYC's promoter is governed by a higher order G-quadruplex (G4) structure in the NHE III1 region. Under a variety of conditions, multiple isoforms have been described to form from the first four continuous guanine runs (G41–4) predominating under the physiologically relevant supercoiled conditions. In the current study, short oligonucleotides complementing the 5′- and 3′-regions flanking the G4 have been connected by an abasic linker to form G4 clamps, varying both linker length and G4 isoform being targeted. Clamp A with an 18 Å linker was found to have marked affinity for its target isomer (G41–4) over the other major structures (G42–5 and G41–5, recognized by clamps B and C, respectively), and to be able to shift equilibrating DNA to foster greater G4 formation. In addition, clamp A, but not B or C, is able to modulate MYC promoter activity with a significant and dose-dependent effect on transcription driven by the Del4 plasmid. This linked clamp-mediated approach to G4 recognition represents a novel therapeutic mechanism with specificity for an individual promoter structure, amenable to a large array of promoters. PMID:27789698

  18. Applications of Dynamic Clamp to Cardiac Arrhythmia Research: Role in Drug Target Discovery and Safety Pharmacology Testing

    Directory of Open Access Journals (Sweden)

    Francis A. Ortega

    2018-01-01

    Full Text Available Dynamic clamp, a hybrid-computational-experimental technique that has been used to elucidate ionic mechanisms underlying cardiac electrophysiology, is emerging as a promising tool in the discovery of potential anti-arrhythmic targets and in pharmacological safety testing. Through the injection of computationally simulated conductances into isolated cardiomyocytes in a real-time continuous loop, dynamic clamp has greatly expanded the capabilities of patch clamp outside traditional static voltage and current protocols. Recent applications include fine manipulation of injected artificial conductances to identify promising drug targets in the prevention of arrhythmia and the direct testing of model-based hypotheses. Furthermore, dynamic clamp has been used to enhance existing experimental models by addressing their intrinsic limitations, which increased predictive power in identifying pro-arrhythmic pharmacological compounds. Here, we review the recent advances of the dynamic clamp technique in cardiac electrophysiology with a focus on its future role in the development of safety testing and discovery of anti-arrhythmic drugs.

  19. Enhanced chromatin accessibility of the dosage compensated Drosophila male X-chromosome requires the CLAMP zinc finger protein.

    Science.gov (United States)

    Urban, Jennifer; Kuzu, Guray; Bowman, Sarah; Scruggs, Benjamin; Henriques, Telmo; Kingston, Robert; Adelman, Karen; Tolstorukov, Michael; Larschan, Erica

    2017-01-01

    The essential process of dosage compensation is required to equalize gene expression of X-chromosome genes between males (XY) and females (XX). In Drosophila, the conserved Male-specific lethal (MSL) histone acetyltransferase complex mediates dosage compensation by increasing transcript levels from genes on the single male X-chromosome approximately two-fold. Consistent with its increased levels of transcription, the male X-chromosome has enhanced chromatin accessibility, distinguishing it from the autosomes. Here, we demonstrate that the non-sex-specific CLAMP (Chromatin-linked adaptor for MSL proteins) zinc finger protein that recognizes GA-rich sequences genome-wide promotes the specialized chromatin environment on the male X-chromosome and can act over long genomic distances (~14 kb). Although MSL complex is required for increasing transcript levels of X-linked genes, it is not required for enhancing global male X-chromosome chromatin accessibility, and instead works cooperatively with CLAMP to facilitate an accessible chromatin configuration at its sites of highest occupancy. Furthermore, CLAMP regulates chromatin structure at strong MSL complex binding sites through promoting recruitment of the Nucleosome Remodeling Factor (NURF) complex. In contrast to the X-chromosome, CLAMP regulates chromatin and gene expression on autosomes through a distinct mechanism that does not involve NURF recruitment. Overall, our results support a model where synergy between a non-sex-specific transcription factor (CLAMP) and a sex-specific cofactor (MSL) creates a specialized chromatin domain on the male X-chromosome.

  20. Linchpin DNA-binding residues serve as go/no-go controls in the replication factor C-catalyzed clamp-loading mechanism.

    Science.gov (United States)

    Liu, Juan; Zhou, Yayan; Hingorani, Manju M

    2017-09-22

    DNA polymerases depend on circular sliding clamps for processive replication. Clamps must be loaded onto primer-template DNA (ptDNA) by clamp loaders that open and close clamps around ptDNA in an ATP-fueled reaction. All clamp loaders share a core structure in which five subunits form a spiral chamber that binds the clamp at its base in a twisted open form and encloses ptDNA within, while binding and hydrolyzing ATP to topologically link the clamp and ptDNA. To understand how clamp loaders perform this complex task, here we focused on conserved arginines that might play a central coordinating role in the mechanism because they can alternately contact ptDNA or Walker B glutamate in the ATPase site and lie close to the clamp loader-clamp-binding interface. We mutated Arg-84, Arg-88, and Arg-101 in the ATPase-active B, C, and D subunits of Saccharomyces cerevisiae replication factor C (RFC) clamp loader, respectively, and assessed the impact on multiple transient events in the reaction: proliferating cell nuclear antigen (PCNA) clamp binding/opening/closure/release, ptDNA binding/release, and ATP hydrolysis/product release. The results show that these arginines relay critical information between the PCNA-binding, DNA-binding, and ATPase sites at all steps of the reaction, particularly at a checkpoint before RFC commits to ATP hydrolysis. Moreover, their actions are subunit-specific with RFC-C Arg-88 serving as an accelerator that enables rapid ATP hydrolysis upon contact with ptDNA and RFC-D Arg-101 serving as a brake that confers specificity for ptDNA as the correct substrate for loading PCNA. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  1. Mycobacterium tuberculosis class II apurinic/apyrimidinic-endonuclease/3'-5' exonuclease III exhibits DNA regulated modes of interaction with the sliding DNA β-clamp.

    Science.gov (United States)

    Khanam, Taran; Rai, Niyati; Ramachandran, Ravishankar

    2015-10-01

    The class-II AP-endonuclease (XthA) acts on abasic sites of damaged DNA in bacterial base excision repair. We identified that the sliding DNA β-clamp forms in vivo and in vitro complexes with XthA in Mycobacterium tuberculosis. A novel 239 QLRFPKK245 motif in the DNA-binding domain of XthA was found to be important for the interactions. Likewise, the peptide binding-groove (PBG) and the C-terminal of β-clamp located on different domains interact with XthA. The β-clamp-XthA complex can be disrupted by clamp binding peptides and also by a specific bacterial clamp inhibitor that binds at the PBG. We also identified that β-clamp stimulates the activities of XthA primarily by increasing its affinity for the substrate and its processivity. Additionally, loading of the β-clamp onto DNA is required for activity stimulation. A reduction in XthA activity stimulation was observed in the presence of β-clamp binding peptides supporting that direct interactions between the proteins are necessary to cause stimulation. Finally, we found that in the absence of DNA, the PBG located on the second domain of the β-clamp is important for interactions with XthA, while the C-terminal domain predominantly mediates functional interactions in the substrate's presence. © 2015 John Wiley & Sons Ltd.

  2. Monitoring of bolt clamping force at high temperatures using metal-packaged regenerated fiber Bragg grating strain sensors

    Science.gov (United States)

    Huang, Yong-Kuo; Tu, Yun; Tu, Shan-Tung

    2017-10-01

    Insufficient or excessive preload has been regarded as the main issue that may endanger the safety of bolted joints operated at high temperatures. A new technique for monitoring the bolt clamping force is thus proposed using metal-packaged regenerated fiber Bragg grating (RFBG) strain sensors spot-welded onto the bolt. The measurement of the clamping force in a bolted steel-steel joint subjected to thermal loading is conducted. The experimental results show that the bolt clamping force can be monitored by the metal-packaged RFBG strain sensor at high temperatures up to 500 °C. This technique provides a new possibility for structural health monitoring of bolted joints operated at high temperatures.

  3. Comparison: Simulation and Experimental Characterisation of an all-Optical Gain-Clamped Erbium-Doped Fibre Amplifier

    Directory of Open Access Journals (Sweden)

    T. Subramaniam M. A. Mahdi P. Poopalan, and H. Ahmad

    2012-10-01

    Full Text Available This paper presents the GC-EDFA characteristics comparison between the simulated results (using EDFA_Design software and experimental results. The comparisons reveal the usefulness of the software in simulating the behaviour of an all-optical GC-EDFA system. Comparisons are made for values obtained from the system operated at high laser power, in order to highlight the differences between the experimental and simulated values. The main objective for this comparison is to prove the capability of the software in simulating the gain-clamped system. Therefore, the software can be used to test new configurations, aimed at improvising current gain-clamped EDFA performances.Keywords optical, gain-clamping, erbium, fibre amplifier, simulation, experiment

  4. Betaalpha-hairpin clamps brace betaalphabeta modules and can make substantive contributions to the stability of TIM barrel proteins.

    Directory of Open Access Journals (Sweden)

    Xiaoyan Yang

    Full Text Available Non-local hydrogen bonding interactions between main chain amide hydrogen atoms and polar side chain acceptors that bracket consecutive betaalpha or alphabeta elements of secondary structure in alphaTS from E. coli, a TIM barrel protein, have previously been found to contribute 4-6 kcal mol(-1 to the stability of the native conformation. Experimental analysis of similar betaalpha-hairpin clamps in a homologous pair of TIM barrel proteins of low sequence identity, IGPS from S. solfataricus and E. coli, reveals that this dramatic enhancement of stability is not unique to alphaTS. A survey of 71 TIM barrel proteins demonstrates a 4-fold symmetry for the placement of betaalpha-hairpin clamps, bracing the fundamental betaalphabeta building block and defining its register in the (betaalpha(8 motif. The preferred sequences and locations of betaalpha-hairpin clamps will enhance structure prediction algorithms and provide a strategy for engineering stability in TIM barrel proteins.

  5. Firing clamp: A novel method for single-trial estimation of excitatory and inhibitory synaptic neuronal conductances

    Directory of Open Access Journals (Sweden)

    Anton eChizhov

    2014-03-01

    Full Text Available Understanding non-stationary neuronal activity as seen in vivo requires estimation of both excitatory and inhibitory synaptic conductances from a single trial of recording. We propose a new intracellular recording method for this purpose called firing clamp. Synaptic conductances are estimated from the characteristics of artificially evoked probe spikes, namely the spike amplitude and the mean subthreshold potential, which are sensitive to both excitatory and inhibitory synaptic input signals. The probe spikes, timed at a fixed rate, are evoked in the dynamic-clamp mode by injected meander-like current steps, with the step duration depending on neuronal membrane voltage. We test the method with perforated-patch recordings from isolated cells stimulated by external application or synaptic release of transmitter, and validate the method with simulations of a biophysically-detailed neuron model. The results are compared with the conductance estimates based on conventional current-clamp recordings.

  6. Patch clamping on plane glass-fabrication of hourglass aperture and high-yield ion channel recording.

    Science.gov (United States)

    Chen, Chang-Yu; Tu, Ting-Yuan; Chen, Chang-Hung; Jong, De-Shien; Wo, Andrew M

    2009-08-21

    Planar patch-clamp has revolutionized ion-channel measurement by eliminating laborious manipulation from the traditional micropipette approach and enabling high throughput. However, low yield in gigaseal formation and/or relatively high cost due to microfabricated processes are two main drawbacks. This paper presents patch clamping on glass substrate-an economical solution without sacrificing gigaseal yield rate. Two-stage CO(2) laser drilling methodology was used to generate an hourglass, funnel-like aperture of a specified diameter with smooth and debris-free surfaces on 150 microm borosilicate cover glass. For 1-3 microm apertures as patch-clamp chips, seal resistance was tested on human embryonic kidney, Chinese hamster ovary, and Jurkat T lymphoma cells with a gigaseal success rate of 62.5%, 43.6% and 66.7% respectively. Results also demonstrated both whole-cell and single channel recording on endogenously expressed ion channels to confirm the capability of different patch configurations.

  7. Robust Clamping Force Control of an Electro-Mechanical Brake System for Application to Commercial City Buses

    Directory of Open Access Journals (Sweden)

    Sangjune Eum

    2017-02-01

    Full Text Available This paper proposes a sensor-less robust force control method for improving the control performance of an electro-mechanical brake (EMB which is applicable to commercial city buses. The EMB generates the accurate clamping force commanded by a driver through an independent motor control at each wheel instead of using existing mechanical components. In general, an EMB undergoes parameter variation and a backdrivability problem. For this reason, the cascade control strategy (e.g., force-position cascade control structure is proposed and the disturbance observer is employed to enhance control robustness against model variations. Additionally, this paper proposed the clamping force estimation method for a sensor-less control, i.e., the clamping force observer (CFO. Finally, in order to confirm the performance and effectiveness of a proposed robust control method, several experiments are performed and analyzed.

  8. State-of-the-art automated patch clamp devices: Heat activation, action potentials and high throughput in ion channel screening

    Directory of Open Access Journals (Sweden)

    Sonja eStoelzle

    2011-11-01

    Full Text Available Ion channels are essential in a wide range of cellular functions and their malfunction underlies many disease states making them important targets in drug discovery. Diverse automated patch clamp systems with high-throughput capabilities are available for drug screening, but there are limitations in the application range. For example, solution exchange time, temperature control and the availability of the current clamp mode can be limiting factors. However, further development of existing devices and introduction of new systems widen the range of possible experiments and increase throughput. Here we introduce new features and platforms that meet the needs of drug discovery researchers and basic researchers alike.The Patchliner is an automated patch clamp system capable of recording up to 8 cells simultaneously with high success rates. Novel features such as temperature control and recordings in the current clamp mode are described here. Standard cell lines and excitable cells such as stem cell-derived cardiomyocytes have been used in the voltage clamp and current clamp modes with the view to finding new drug candidates and safety testing methods in a more physiologically relevant environment. The SyncroPatch 96, is a screening platform capable of recording from 96 cells in parallel and offers a throughput of 5000 data points per day. Full dose response curves can be acquired from individual cells reducing the cost per data point. The system is an ideal tool for secondary screening efforts and for safety testing on ligand- and voltage-gated ion channels.The Patchliner and SyncroPatch 96 are ideal platforms for drug discovery, ion channel research and safety testing, combining long awaited features such as parallel action potential recordings and temperature control with extensively increased throughput.

  9. Ramp-incremented and RPE-clamped test protocols elicit similar VO2max values in trained cyclists.

    Science.gov (United States)

    Straub, Allison M; Midgley, Adrian W; Zavorsky, Gerald S; Hillman, Angela R

    2014-08-01

    The present study compared the efficacy of ramp incremented and ratings of perceived exertion (RPE)-clamped test protocols for eliciting maximal oxygen uptake (VO2max). Sixteen trained cyclists (age 34 ± 7 years) performed a ramp-incremented protocol and an RPE-clamped protocol 1 week apart in a randomized, counterbalanced order. The RPE-clamped protocol consisted of five, 2-min stages where subjects self-selected work rate and pedal cadence to maintain the prescribed RPE. After completing both test protocols subjects were asked which they preferred. The mean ± SD test time of 568 ± 72 s in the ramp protocol was not significantly different to the 600 ± 0 s in the RPE-clamped protocol (mean difference = 32 s; p = 0.09), or was the VO2max of 3.86 ± 0.73 L min(-1) in the ramp protocol significantly different to the 3.87 ± 0.72 L min(-1) in the RPE-clamped protocol (mean difference = 0.002 L min(-1); p = 0.97). Furthermore, no significant differences were observed for peak power output (p = 0.21), maximal minute ventilation (p = 0.97), maximal respiratory exchange ratio (p = 0.09), maximal heart rate (p = 0.51), and post-test blood lactate concentration (p = 0.58). The VO2max attained in the preferred protocol was significantly higher than the non-preferred protocol (mean difference = 0.14 L min(-1); p = 0.03). The RPE-clamped test protocol was as effective as the ramp-incremented protocol for eliciting VO2max and could be considered as a valid alternative protocol, particularly where a fixed test duration is desirable.

  10. Research and development of a self-centering clamping device for deep-water multifunctional pipeline repair machinery

    Directory of Open Access Journals (Sweden)

    Liquan Wang

    2016-01-01

    Full Text Available When multifunctional pipeline repair machinery (MPRM is used in the deep sea area, it is difficult to grip the pipeline and ensure concentricity between the cutter heads and the pipeline during its operation. In view of this, a new system of two-arm holding self-centering pipeline clamping device was proposed. The system is composed of two groups of parallelogram double-rocker mechanism and cranking block mechanism which are symmetrically distributed on the frame. The geometric parameter solutions of the clamping device were analyzed with motion and transmission as the constraints. A mechanical model was established to associate the friction torque of clamping points with the driving force. Clamping device and machinery were designed and manufactured for the Ø304.8–457.2 mm pipelines used in this test. ADAMS simulation experiments were conducted underwater, and the cutting and beveling tests were carried out onshore. The following results are achieved. First, the smaller the pipe diameter, the smaller the transmission angle of the oscillating slider mechanism; the longer the hydraulic cylinder stroke, the greater the transmission angle of the double rocker mechanism. Second, the driving force of the clamping device increases with the increase of the pipe diameter. When the diameter reaches 457.2 mm, the hydraulic cylinder driving force of the clamping device should be greater than 10219 N. Third, the feed rate of the cutters increases suddenly due to the slight shaking of the machinery which occurs at the beginning of the pipe cutting, so it is necessary to adopt a small feed rate. And fourth, onshore experiment results agree well with the theoretical design and simulation results, proving the rationality of the system. The research results in this paper provide technical basis for the research and development of similar engineering prototypes.

  11. Structure of the sliding clamp from the fungal pathogen Aspergillus fumigatus (AfumPCNA) and interactions with Human p21.

    Science.gov (United States)

    Marshall, Andrew C; Kroker, Alice J; Murray, Lauren A M; Gronthos, Kahlia; Rajapaksha, Harinda; Wegener, Kate L; Bruning, John B

    2017-03-01

    The fungal pathogen Aspergillus fumigatus has been implicated in a drastic increase in life-threatening infections over the past decade. However, compared to other microbial pathogens, little is known about the essential molecular processes of this organism. One such fundamental process is DNA replication. The protein responsible for ensuring processive DNA replication is PCNA (proliferating cell nuclear antigen, also known as the sliding clamp), which clamps the replicative polymerase to DNA. Here we present the first crystal structure of a sliding clamp from a pathogenic fungus (A. fumigatus), at 2.6Å. Surprisingly, the structure bears more similarity to the human sliding clamp than other available fungal sliding clamps. Reflecting this, fluorescence polarization experiments demonstrated that AfumPCNA interacts with the PCNA-interacting protein (PIP-box) motif of human p21 with an affinity (Kd ) of 3.1 μm. Molecular dynamics simulations were carried out to better understand how AfumPCNA interacts with human p21. These simulations revealed that the PIP-box bound to AfuPCNA forms a secondary structure similar to that observed in the human complex, with a central 310 helix contacting the hydrophobic surface pocket of AfumPCNA as well as a β-strand that forms an antiparallel sheet with the AfumPCNA surface. Differences in the 310 helix interaction with PCNA, attributed to residue Thr131 of AfumPCNA, and a less stable β-strand formation, attributed to residues Gln123 and His125 of AfumPCNA, are likely causes of the over 10-fold lower affinity of the p21 PIP-box for AfumPCNA as compared to hPCNA. The atomic coordinates and structure factors for the Aspergillus fumigatus sliding clamp can be found in the RCSB Protein Data Bank (http://www.rcsb.org) under the accession code 5TUP. © 2017 Federation of European Biochemical Societies.

  12. Novel description of ionic currents recorded with the action potential clamp technique: application to excitatory currents in suprachiasmatic nucleus neurons.

    Science.gov (United States)

    Clay, John R

    2015-07-01

    The traditional method of recording ionic currents in neurons has been with voltage-clamp steps. Other waveforms such as action potentials (APs) can be used. The AP clamp method reveals contributions of ionic currents that underlie excitability during an AP (Bean BP. Nat Rev Neurosci 8: 451-465, 2007). A novel usage of the method is described in this report. An experimental recording of an AP from the literature is digitized and applied computationally to models of ionic currents. These results are compared with experimental AP-clamp recordings for model verification or, if need be, alterations to the model. The method is applied to the tetrodotoxin-sensitive sodium ion current, INa, and the calcium ion current, ICa, from suprachiasmatic nucleus (SCN) neurons (Jackson AC, Yao GL, Bean BP. J Neurosci 24: 7985-7998, 2004). The latter group reported voltage-step and AP-clamp results for both components. A model of INa is constructed from their voltage-step results. The AP clamp computational methodology applied to that model compares favorably with experiment, other than a modest discrepancy close to the peak of the AP that has not yet been resolved. A model of ICa was constructed from both voltage-step and AP-clamp results of this component. The model employs the Goldman-Hodgkin-Katz equation for the current-voltage relation rather than the traditional linear dependence of this aspect of the model on the Ca(2+) driving force. The long-term goal of this work is a mathematical model of the SCN AP. The method is general. It can be applied to any excitable cell.

  13. Cascading MutS and MutL sliding clamps control DNA diffusion to activate mismatch repair.

    Science.gov (United States)

    Liu, Jiaquan; Hanne, Jeungphill; Britton, Brooke M; Bennett, Jared; Kim, Daehyung; Lee, Jong-Bong; Fishel, Richard

    2016-11-24

    Mismatched nucleotides arise from polymerase misincorporation errors, recombination between heteroallelic parents and chemical or physical DNA damage. Highly conserved MutS (MSH) and MutL (MLH/PMS) homologues initiate mismatch repair and, in higher eukaryotes, act as DNA damage sensors that can trigger apoptosis. Defects in human mismatch repair genes cause Lynch syndrome or hereditary non-polyposis colorectal cancer and 10-40% of related sporadic tumours. However, the collaborative mechanics of MSH and MLH/PMS proteins have not been resolved in any organism. We visualized Escherichia coli (Ec) ensemble mismatch repair and confirmed that EcMutS mismatch recognition results in the formation of stable ATP-bound sliding clamps that randomly diffuse along the DNA with intermittent backbone contact. The EcMutS sliding clamps act as a platform to recruit EcMutL onto the mismatched DNA, forming an EcMutS-EcMutL search complex that then closely follows the DNA backbone. ATP binding by EcMutL establishes a second long-lived DNA clamp that oscillates between the principal EcMutS-EcMutL search complex and unrestricted EcMutS and EcMutL sliding clamps. The EcMutH endonuclease that targets mismatch repair excision only binds clamped EcMutL, increasing its DNA association kinetics by more than 1,000-fold. The assembly of an EcMutS-EcMutL-EcMutH search complex illustrates how sequential stable sliding clamps can modulate one-dimensional diffusion mechanics along the DNA to direct mismatch repair.

  14. A scanning force microscope for simultaneous force and patch-clamp measurements on living cell tissues

    Science.gov (United States)

    Langer, M. G.; Öffner, W.; Wittmann, H.; Flösser, H.; Schaar, H.; Häberle, W.; Pralle, A.; Ruppersberg, J. P.; Hörber, J. K. H.

    1997-06-01

    For the investigation of mechanosensitive ion channels of living cells it is of great interest to apply very local forces in the piconewton range and to measure, simultaneously, ion currents down to 1 pA. Scanning force microscopy (SFM) is a suitable technique, that allows the application of such small forces with a lateral resolution in the range of 10 nm. We developed a novel type of experimental setup, because no existing SFM, home built or commercial, allows a simultaneous investigation of ion currents and mechanical properties of living cells. The construction consists of a SFM that is combined with an upright infrared differential interference contrast (DIC) video microscope and a conventional patch-clamp setup. Instead of the object, the force sensor is scanned to prevent relative movements between the patch pipette and the patched cell. The deflection of the SFM cantilever is detected with the so-called optical deflection method through the objective of the optical microscope. In opposite to common optical setups the laser beam was not focused on the force sensor. The presented optic creates a parallel laser beam between the objective and the SFM cantilever, which allows a vertical displacement of the sensor without any changes of the detector signal. For the three-dimensional positioning of the specimen chamber a two-axis translation stage including a vertical piezoelectric translation device was developed. The SFM tip is fixed on a combined lateral and vertical translation stage including a piezoelectric tube scanner for three-dimensional fine positioning. Thus the instrument enables an easy approach of the SFM tip to any optically identified cell structure. The head stage of the patch-clamp electronics and the patch pipette are directly fixed on the specimen stage. This prevents relative movements between patched cells and patch pipette during the approach to the SFM tip. The three-axis positioning of the patch pipette is done by a compact hydraulic

  15. An Experimental Method for Measuring the Clamping Force in Double Lap Simple Bolted and Hybrid (Bolted-Bonded) Joints

    OpenAIRE

    Goldarag, Firooz Esmaeili; Barzegar, Sajjad; Babaei, Alireza

    2015-01-01

    In this research, an experimental method for measuring the clamping force as a result of tightening torque in double lap simple bolted and hybrid (bolted-bonded) joints is proposed. Two types of joints, i.e. double lap simple and hybrid (bolted-bonded) joints were prepared for testing. In order to measure the clamping force or pretension resulting from the tightening torque at different applied torques, for both types of joints, a special experimental method was designed using a steel bush th...

  16. Transistor Clamped Five-Level Inverter using Non-Inverting Double Reference Single Carrier PWM Technique for photovoltaic applications

    DEFF Research Database (Denmark)

    Bhaskar, Mahajan Sagar; Padmanaban, Sanjeevikumar; Fedák, Viliam

    2017-01-01

    This treatise deals with Transistor Clamped Five Level Inverter using Non-Inverting Double Reference Single Carrier PWM (NIDRSC PWM) Technique. Conventional or two level inverter have drawbacks like: i) Requirement of fast switching devices, ii) Very high dv/dt, iii) High Electromagnetic Interfer......This treatise deals with Transistor Clamped Five Level Inverter using Non-Inverting Double Reference Single Carrier PWM (NIDRSC PWM) Technique. Conventional or two level inverter have drawbacks like: i) Requirement of fast switching devices, ii) Very high dv/dt, iii) High Electromagnetic...

  17. Impact of Individual Proliferating Cell Nuclear Antigen-DNA Contacts on Clamp Loading and Function on DNA*

    Science.gov (United States)

    Zhou, Yayan; Hingorani, Manju M.

    2012-01-01

    Ring-shaped clamp proteins encircle DNA and affect the work of many proteins, notably processive replication by DNA polymerases. Crystal structures of clamps show several cationic residues inside the ring, and in a co-crystal of Escherichia coli β clamp-DNA, they directly contact the tilted duplex passing through (Georgescu, R. E., Kim, S. S., Yurieva, O., Kuriyan, J., Kong, X. P., and O'Donnell, M. (2008) Structure of a sliding clamp on DNA. Cell 132, 43–54). To investigate the role of these contacts in reactions involving circular clamps, we examined single arginine/lysine mutants of Saccharomyces cerevisiae proliferating cell nuclear antigen (PCNA) in replication factor C (RFC)-catalyzed loading of the clamp onto primer template DNA (ptDNA). Previous kinetic analysis has shown that ptDNA entry inside an ATP-activated RFC-PCNA complex accelerates clamp opening and ATP hydrolysis, which is followed by slow PCNA closure around DNA and product dissociation. Here we directly measured multiple steps in the reaction (PCNA opening, ptDNA binding, PCNA closure, phosphate release, and complex dissociation) to determine whether mutation of PCNA residues Arg-14, Lys-20, Arg-80, Lys-146, Arg-149, or Lys-217 to alanine affects the reaction mechanism. Contrary to earlier steady state analysis of these mutants (McNally, R., Bowman, G. D., Goedken, E. R., O'Donnell, M., and Kuriyan, J. (2010) Analysis of the role of PCNA-DNA contacts during clamp loading. BMC Struct. Biol. 10, 3), our pre-steady state data show that loss of single cationic residues can alter the rates of all DNA-linked steps in the reaction, as well as movement of PCNA on DNA. These results explain an earlier finding that individual arginines and lysines inside human PCNA are essential for polymerase δ processivity (Fukuda, K., Morioka, H., Imajou, S., Ikeda, S., Ohtsuka, E., and Tsurimoto, T. (1995) Structure-function relationship of the eukaryotic DNA replication factor, proliferating cell nuclear antigen

  18. Sex-dependent reductions in high-molecular weight adiponectin during acute hyperinsulinemia are prevented with endurance training in older females.

    Science.gov (United States)

    Consitt, Leslie A; Saxena, Gunjan; Schaefer, Megan

    2018-02-07

    The high molecular weight (HMW) adiponectin isoform is considered the active form of adiponectin and is linked to insulin sensitivity and the reduced risk of developing cardiovascular disease. The purpose of the first study was to determine the effects of age and sex on the plasma HMW adiponectin response to acute hyperinsulinemia, and secondly determine if either endurance or resistance exercise training could affect this response. Twenty-six, healthy males (19-84 years) and twenty-six, healthy females (18-76 years) were recruited and matched for BMI to examine the effects of sex and age on the plasma adiponectin response to a 2 hour hyperinsulinemic-euglycemic clamp. To examine the effects of exercise training, a subgroup of young (55 years) individuals were randomized into a 12-week endurance or resistance training program and had their adiponectin response to hyperinsulinemia measured before and after training. HMW and total adiponectin were measured by ELISA. In response to hyperinsulinemia, plasma HMW adiponectin decreased in females (-9%, Pendurance training, the response of plasma HMW adiponectin to hyperinsulinemia increased in older females (36%, Ptraining had no effect on the plasma adiponectin response to hyperinsulinemia. Despite no age or sex differences at baseline, skeletal muscle AdipoR1 increased in response to endurance training (~120%, Ptraining (~38%, Pendurance training protects older females against the hyperinsulinemic inhibition of plasma HMW adiponectin, which could promote healthy aging. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  19. Hyperinsulinism and polycystic ovary syndrome (PCOS): role of insulin clearance.

    Science.gov (United States)

    Amato, M C; Vesco, R; Vigneri, E; Ciresi, A; Giordano, C

    2015-12-01

    Insulin resistance and compensatory hyperinsulinism are the predominant metabolic defects in polycystic ovary syndrome (PCOS). However, hyperinsulinism, as well as being compensatory, can also express a condition of reduced insulin clearance. Our aim was to evaluate the differences in insulin action and metabolism between women with PCOS (with normal glucose tolerance) and age- and BMI-matched women with prediabetes (without hyperandrogenism and ovulatory disorders). 22 women with PCOS and 21 age/BMI-matched women with prediabetes were subjected to a Hyperinsulinemic-euglycemic clamp and an Oral Glucose tolerance Test (OGTT). Insulin sensitivity was assessed by the glucose infusion rate during clamp (M value); insulin secretion by Insulinogenic index, Oral Disposition Index (DIo) and AUC(2h-insulin) during OGTT; and insulin clearance by the metabolic clearance rate of insulin (MCRI) during clamp. Women with PCOS showed significantly higher levels of AUC(2h-insulin) (p PCOS [420 (IQR 227-588) vs. 743 (IQR 597-888) ml m(-2) min(-1): p PCOS group, a strong independent inverse correlation was only observed between MCRI and AUC(2h-insulin) (PCOS: β:-0.878; p PCOS there is peripheral insulin sensitivity similar to that of women with prediabetes. What sets PCOS apart is the hyperinsulinism, today still simplistically defined "compensatory"; actually this is mainly related to decreased insulin clearance whose specific causes and dynamics have yet to be clarified.

  20. Insulin resistance of protein anabolism accompanies that of glucose metabolism in lean, glucose-tolerant offspring of persons with type 2 diabetes.

    Science.gov (United States)

    Burgos, Sergio A; Chandurkar, Vikram; Tsoukas, Michael A; Chevalier, Stéphanie; Morais, José A; Lamarche, Marie; Marliss, Errol B

    2016-01-01

    To test whether protein anabolic resistance is an early defect in type 2 diabetes (T2D). Seven lean, normoglycemic T2D offspring (T2D-O) and eight matched participants without family history (controls; C) underwent a 3-hour hyperinsulinemic (40 mU/m2/min), euglycemic (5.5 mmol/L) and isoaminoacidemic clamp. Whole-body glucose and protein kinetics were measured with d-[3-3H]glucose and l-[l-13C]leucine, respectively. Plasma amino acids were measured by liquid chromatography-tandem mass spectrometry. Fasting glycemia and glucose kinetic variables did not differ between groups. Clamp decreases in glucose rate of appearance were not different, but rate of disappearance increased 29% less in T2D-O, to a significantly lower rate. Fasting leucine was higher in T2D-O, but kinetics did not differ. Clamp increases in leucine oxidation and decreases in endogenous rate of appearance (protein breakdown) were equal, but in T2D-O, non-oxidative rate of disappearance (protein synthesis) did not increase and net balance (synthesis-breakdown) did not become positive as in C. Resistance of whole-body protein anabolism (synthesis and net balance) accompanies resistance of glucose uptake in T2D-O. Mechanisms responsible, possible roles in the increased risk of developing diabetes, and its potential impact on long-term protein balance require definition.