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At least 379 records · Page 21Linked to original sources
PubMed1997

Urinary excretion of adrenaline and noradrenaline during hypoglycaemic clamp in diabetic and non-diabetic adolescents.

The urinary excretion of non-conjugated adrenaline and noradrenaline during hypoglycaemic clamp was investigated in 16 diabetic and 15 non-diabetic adolescents. In the diabetic adolescents, the mean excretion of adrenaline was approximately nine times higher in clamp urine than in morning urine (p < 0.0001). In the non-diabetic adolescents, the mean excretion of adrenaline was 20-30 times higher in clamp urine than in morning urine (p < 0.0001). Plasma catecholamines were measured, and the area under the plasma concentration-time curve (AUC) and renal clearance of catecholamines were calculated. There was a linear correlation between the AUC and the urinary excretion of adrenaline (r = 0.57, p = 0.001) and noradrenaline (r = 0.49, p = 0.006) during the hypoglycaemic clamp. The renal clearance of adrenaline and noradrenaline did not differ between the diabetic and non-diabetic subjects. In the diabetic group, there was a positive correlation between HbA1c and the adrenaline excretion in clamp urine (r = 0.60, p = 0.015). The eight diabetic subjects with HbA1c < 10% had lower adrenaline excretion in clamp urine than the subjects with HbA1c > 10% and the non-diabetic subjects (all p values < 0.05). We conclude that there is a marked increase in urinary adrenaline excretion during hypoglycaemia in diabetic and non-diabetic adolescents. The increase in adrenaline excretion during hypoglycaemia was attenuated in the better-controlled diabetic adolescents in our study.

Adolescent↗
PubMedDate not supplied

Comparison of several formulas to assess insulin action in the fasting state with the hyperglycemic-hyperinsulinemic clamp technique in healthy individuals.

BACKGROUND: The aim of this study was to correlate several formulas to assess insulin action in the fasting state with the hyperglycemic-hyperinsulinemic clamp method in healthy individuals. METHODS: A cross-sectional study was carried out in 33 healthy, non-obese volunteers. A hyperglycemic-hyperinsulinemic clamp technique was performed and the following formulas to assess insulin action in the fasting state were calculated: reciprocal insulin, glucose to insulin ratio, homeostasis model analysis, Raynaud index, Belfiore index, reciprocal of fasting insulin resistance index and quantitative insulin sensitivity check index. Pearson correlation was used to correlate the above mentioned formulas with the clamp method. RESULTS: There were significant correlations between results of the several formulas and those obtained with the clamp technique. The reciprocal of insulin index had better correlation with the clamp method. CONCLUSIONS: All formulas evaluated to assess insulin action in the fasting state had a significant correlation with the hyperglycemic clamp technique.

Adolescent↗
PubMed1991

A new glucose clamp algorithm: clinical validation.

A new glucose clamp technique for in vivo studies of insulin sensitivity was validated clinically. Eighteen patients (10 males, 8 females, age 35-80 years, body mass index 34.6-17.04) were connected to a computer-assisted artificial pancreas "Betalike R", using a new algorithm based on a "minimal model", to carry out the glucose clamp technique automatically and especially to overcome the well-known problems of its priming phase. We performed the euglycemic hyperinsulinemic clamp in four patients and the hyperglycemic hyperinsulinemic clamp in 14. In one patient both clamps were done. The mean priming time to reach steady-state glycemia was 20 min. Plasma insulin concentrations were measured every 20 min. This new automatic glucose clamp technique enables the priming phase to be run without any significant overshoot, and accidental variations of glycemia in steady state were reduced to a minimum. The system showed satisfactory safety and stability in controlling the patient's glycemia and assured high speed of the priming phase.

Adult↗
PubMed1989

The derivation of a minimal model insulin sensitivity index from euglycaemic clamps in man.

We describe a computer program based on the minimal model of Bergman et al., which has been used to control euglycaemic hyperinsulinaemic clamps and to derive the insulin sensitivity index (Si) from clamp data. Clamps were undertaken on 33 subjects with a range of insulin resistance. The steady state glucose had an acceptable coefficient of variation (c.v.) and was close to the desired clamp level. The c.v. of the steady state glucose infusion rate was smaller than for other clamp programmes. The Si was correlated with metabolic clearance rate for glucose (MCR) divided by the change in insulin concentration. Our results suggest that the assumptions of the minimal model hold true in the euglycaemic hyper-insulinaemic situation and demonstrate that a minimal-model-derived program can achieve acceptable clamping.

Algorithms↗
PubMed1984

Renal and splanchnic circulation during infrarenal aortic cross-clamping.

The effect of infrarenal cross-clamping of the aorta on regional splanchnic and renal circulations was studied in seven dogs. Regional blood flow was determined with differentially labeled microspheres (9 and 15 micron in diameter) that were injected simultaneously into the left atrium. Blood flow was measured 30 minutes after surgical preparation was completed (stage I), 20 minutes after infrarenal aortic cross-clamping (stage II), and 20 minutes after supplemented sodium nitroprusside infusion (stage III). Infrarenal aortic cross-clamping was accompanied by a slight increase in the cardiac output (CO) without significant changes in mean arterial pressure (MAP). Blood flow through the gut, hepatic artery, and cortical layer of the kidneys, as determined with 15-micron spheres, was not changed. Nonentrapment of 9-micron spheres in the gut and renal cortex was increased substantially. Blood flow through the juxtamedullary layer of the kidneys was increased. Sodium nitroprusside supplementation decreased MAP by 30%; CO values returned to baseline level. Hepatic artery blood flow, compared with both baseline values and values during aortic cross-clamping, increased significantly. Blood flow, determined with 15-micron spheres, through the gut and renal cortex did not change, and nonentrapment of 9-micron spheres decreased to baseline values. The data suggest a certain shift of blood flow to the juxtamedullary layer of the kidneys during aortic cross-clamping and normalization of intrarenal blood flow distribution during supplemented sodium nitroprusside infusion. Controlled vasoplegia with sodium nitroprusside may help modify peripheral circulatory disturbances in the kidneys and splanchnic system during infrarenal aortic cross-clamping.

Animals↗
PubMed1996

Comparison of skeletal muscle microcirculation between clamp ischemia and microsurgical ischemia.

We compared ischemia reperfusion injury-associated vasospasm and perfused capillary density (PCD) at the microcirculatory level between clamp ischemia and microsurgical ischemia in rat skeletal muscle. Rat cremaster muscle was prepared as an island flap, attached only with pudic-epigastric vessels branching from external iliac vessels. Two types of ischemia, with clamping only or with microvascular anastomosis, were applied at the external iliac vessels for 2 hours followed by 1-hour reperfusion before in vivo microscopic examination for hemodynamic changes. At the end of observation, small segments of the vessels at the clamping site and microsurgical anastomoses site were also harvested for histological examination. It was found that the first- and second-order arterioles had about 12-15% diameter reductions in both groups, whereas diameter reductions of the third-order arterioles were up to 37.8% in the microsurgical ischemia group, much greater than that in the clamp ischemia group (2.3%). There was also no significant difference in PCD reduction between the two groups, although the red blood cell velocity was much slower in the microsurgical ischemia group. Histological examination of the anastomosis site showed massive accumulation of polymorphonuclear neutrophils on the venous endothelium. These results suggested a different degree of endothelial damage and local leukocyte activation between microsurgical ischemia and clamp ischemia. Therefore, we conclude that clamp ischemia cannot replace microsurgical ischemia for studying microcirculatory changes in free tissue transfer.

Anastomosis, Surgical↗
PubMed1991

Prolonged intermittent clamping of the portal triad during hepatectomy.

From 112 consecutive hepatectomies for malignant tumours performed with intermittent portal triad clamping, we have retrospectively selected the 20 cases in which clamping exceeded 90 min. Intermittent portal clamping of prolonged duration was used because of abnormal liver texture in 13 cases (mainly patients who had received intra-arterial chemotherapy) and/or because of technically difficult hepatectomy. The mean(s.d.) duration of intermittent portal clamping was 109(18) min and in two cases it exceeded 140 min (148 and 150 min). There was no postoperative mortality and the rate of postoperative morbidity was 35 per cent. Postoperative changes in biochemical liver tests were not major and transient hepatic failure occurred in only one patient following subtotal resection of the liver. We conclude that intermittent portal clamping is a useful manoeuvre in partial hepatectomy when resection is difficult or prolonged, or when the liver parenchyma is abnormal. Such clamping may be used for longer than 120 min without major complications.

Adult↗
PubMed1984

A new microvascular clamp for the production of experimental ischemia.

A new microvascular clamp was developed from inexpensive and readily available materials. This clamp, when properly applied, can provide many hours of vessel occlusion with minimal damage to the vessel intima. This clamp has important applications in producing prolonged experimental ischemia without significant vessel damage and thrombosis. This clamp produced less vessel damage when compared with a standard microvascular clamp for prolonged periods of clamping.

Animals↗
PubMed1996

Fluorescence monitoring of T4 polymerase holoenzyme accessory protein interactions during loading of the sliding clamp onto the template-primer junction.

Assembly of the T4 polymerase holoenzyme requires coordinated interactions among the core polymerase and the clamp loader (gp44/62) and sliding clamp (gp45) accessory proteins. Here we describe the creation of a mutant of gp45 that can be uniquely modified by fluorescent probes within each protein monomer at a site-specific cysteine residue. The fluorescently labeled gp45 was shown to have the same biological activity as the wild-type protein. These strike "labeled" gp45 adducts were then used in steady-state and fluorescence polarization studies to monitor the interaction of gp45 with the gp44/62 clamp-loading complex and template-primer DNA in the presence and absence of ATP, and of the non-hydrolyzable analog, adenosine 5'-O-(3-thiotriphosphate). We find that a complex of ATP-activated gp44/62 with appropriately labeled gp45 shows significant fluorescent enhancement (and an increase in fluorescent anisotropy), that can be partially reversed by interaction with template-primer DNA. Fluorescence-monitored binding curves between gp45 and ATP-activated gp44/62 reveal that the two protein complexes bind with a 1:1 stoichiometry. Analysis shows that these methods can be used to follow the ATP-driven loading of gp45 onto the template-primer by the gp44/62 clamp-loading complex, and in combination with the kinetic data presented in the companion article, provide insight into the rate-limiting steps during clamp assembly on template-primer DNA. A reaction pathway for this processivity clamp-loading process is proposed.

Adenosine Triphosphatases↗
PubMed2001

Locking the ATP-operated clamp of DNA gyrase: probing the mechanism of strand passage.

DNA gyrase catalyses DNA supercoiling by passing one segment of DNA (the T segment) through another (the G segment) in a reaction coupled to the binding and hydrolysis of ATP. The N-terminal domains of the gyrase B dimer constitute an ATP-operated clamp that is proposed to capture the T segment during the DNA supercoiling reaction. We have locked this clamp in the closed conformation using the non-hydrolysable ATP analogue ADPNP (5'-adenylyl beta,gamma-imidodiphosphate). The clamp-locked enzyme is able to bind and cleave DNA, albeit at a reduced level. Although the locked enzyme is not capable of carrying out DNA supercoiling, it can catalyse limited DNA relaxation, consistent with the ability to complete one strand passage event per enzyme molecule via entry of the T segment through the exit gate of the enzyme. The DNA-protein complex of the clamp-locked enzyme has a conformation that differs from the normal positively wrapped conformation of the gyrase-DNA complex. These experiments confirm the role of the ATP-operated clamp in the strand-passage reactions of gyrase and suggest a model for the interaction of DNA with gyrase in which a conformation with the T segment in equilibrium across the DNA gate can be achieved via T-segment entry through the ATP-operated clamp or through the exit gate.

Adenosine Triphosphate↗
PubMed1996

Infrarenal aortic clamp hypertension is exacerbated by baroreceptor blockade.

Aortic clamp-induced hypertension has long been implicated in the cardiovascular mortality and morbidity following infrarenal aortic operations. We studied the physiologic mechanisms leading to clamp-induced hypertension. Mean arterial pressure (MAP), cardiac output, heart rate, and left ventricular pressure were measured in alpha-chloralose-anesthetized dogs. Animals received alpha, beta, both alpha and beta, or no adrenergic blockade (n = 3, 4, 12 and 7, respectively). The infrarenal aorta was clamped following ligation of the infrarenal collateral vessels (lumbar, circumflex iliac, and tail arteries). Statistical analysis used paired t tests within groups, and ANOVA and unpaired t tests between groups, with Bonferroni's correction as indicated. Following placement of the clamp, MAP increased immediately in all groups, with magnitude of the increase related to the extent of adrenergic blockade. MAP increased 5.6 +/- 0.8 mm Hg with no blockade (P = 0.0005), 6.7 +/- 0.8 mm Hg with alpha blockade (P = 0.0153), 15 +/- 3.1 mm Hg with beta blockade (P = 0.0163), and 16.7 +/- 1.3 mm Hg with combined alpha and beta blockade (P < 0.0001). The increase in MAP immediately following infrarenal aortic clamping was most pronounced with combined alpha and beta blockade. We suggest that acute intraoperative hypertension associated with infrarenal aortic clamping is caused by the attenuation of compensatory baroreceptor reflex mechanisms.

Adrenergic beta-Antagonists↗
PubMed1991

An inexpensive inverted microscope for patch-clamp and other electrophysiological studies at the cellular level.

The popularization of the patch-clamp technique has increased the demand for inverted light microscopes that allow the optimal or almost free movement of patch-clamp pipettes and their support drives. However, commercially available models of inverted microscopes have not been specifically designed for this line of research and, as a consequence, patch-clamp pipette movements are restricted by the small space available between the sample, and the light source and its modulating attachments. This paper provides the details for the construction of a relatively inexpensive inverted microscope that meets the specifications required for patch-clamp and other electrophysiological investigations at the cellular level. The microscope allows the free positioning of the conventional probes for patch-clamp, microelectrode amplifiers, and other micromanipulator probes and attachments. The construction of the microscope is simple and, therefore, since it is relatively inexpensive, the microscope may be easily upgraded in many ways for special purposes, including special optical effects. Finally, although the instrument was developed for patch-clamp and classical electrophysiological studies, it may be used in other types of investigations where freedom of microtool movement is imperative, such as in microsurgery applications.

Electrophysiology↗
PubMed1992

Pressure-clamp: a method for rapid step perturbation of mechanosensitive channels.

Here we describe a pressure-clamp method for applying suction or pressure steps to membrane patches in order to study the activation, adaptation and relaxation characteristics of mechanosensitive (MS) channels. A description is given of the mechanical arrangement of the pressure clamp which involves a balance between negative (suction) and positive pressures. The electronic circuitry of the feedback control is described. We also describe the optimal time response (approximately 10 ms) of the pressure-clamp, the amplitude of pressure resolution (0.2-0.5 mmHg; 27-67 Pa) and the factors influencing these parameters. We illustrate the applications of the clamp on the Xenopus oocyte and cultured skeletal myotubes from dystrophic mouse (mdx) muscle, both of which express MS channels. Studies with pressure/suction pulses indicate that in both muscles and oocytes MS channel activity displays adaptation. The ability to study current relaxations following step changes in pressure/suction using the pressure-clamp in combination with patch-clamp techniques provides the opportunity for analysis of the time, voltage and pressure dependence of the opening and closing of MS channels.

Animals↗
PubMed1989

Versatile supplement device with remote control for the control of patch clamp experiments.

A versatile device for a patch-clamp amplifier is described. This device contains: (i) an acoustic indicator to monitor the input resistance of the patch pipette, which is used in search-mode to indicate the formation of seals; (ii) two pulse generators; and (iii) a staircase generator to produce various pulse and voltage step programs; (iv) a low-pass filter which is used to filter the output of the patch clamp amplifier; and (v) a remote control which is used to control the entire patch clamp experiment. This remote control is used to switch between search-, current clamp-, and voltage clamp-mode, to activate the respective stimulus potential programs, and to control the tape recorder. This electronic device can be easily connected to patch clamp amplifiers.

Animals↗
PubMed1983

Modification by preoperative beta-blockade of the renin response to infrarenal aortic cross-clamping.

The activity of the renin-angiotensin system was measured before, during, and after infrarenal aortic cross-clamping in 13 patients. Five of the patients studied were taking propranolol preoperatively and formed a subgroup. Intraoperative blood loss, volume of crystalloid and colloid infused, haemodynamic parameters and urine output were similar for the two groups. In eight patients who were not taking propranolol mean plasma renin activity was 2.24 ng . ml-1 . hr-1 prior to induction, 3.78 ng . ml-1 . hr-1 during surgery prior to cross-clamping and 4.42 ng X ml-1 X hr-1 15 minutes after the aorta was cross-clamped (increases not statistically significant). Mean plasma renin activity measured ten minutes prior to release of the cross-clamp (5.02 ng X ml-1 . hr-1), 15 minutes after clamp release (5.47 ng X ml-1 X hr-1), and 30 minutes after reaching the recovery room (5.84 ng X ml-1 X hr-1) were significantly greater than preinduction levels. Four patients developed postoperative hypertension (mean blood pressure greater than 120 mmHg); there was not a correlation between the elevated plasma renin activity observed postoperatively and the occurrence of postoperative hypertension. The five patients taking propranolol had a markedly attenuated renin activity response during and after surgery; the mean plasma renin activity was less than 1.5 ng X ml-1 X hr-1 at all sampling times. Two of these five patients did develop postoperative hypertension. It is concluded that surgery involving infrarenal aortic cross-clamping is associated with increased plasma renin activity with peak levels occurring postoperatively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed2005

Reduction of abdominal wall blood flow by clamping or carbon dioxide insufflation increases tumor growth in the abdominal wall: an experimental study in rats.

BACKGROUND: We have previously demonstrated that there is a reduction of blood flow in the abdominal wall in rats insufflated with air concomitant with an increase in tumor growth. The present study was designed to examine whether a reduction of blood flow achieved by clamping or insufflation with carbon dioxide (CO(2)) would increase tumor growth in the abdominal wall. METHODS: In the first part of the experiments, laser Doppler blood flow of both rectus muscles was measured in 16 Wistar Fu rats. The left rectus muscle was clamped to reduce blood flow, and 5 x 10(4) adenocarcinoma cells were injected into both rectus muscles. Clamping was maintained for 45 min. In the second part, 22 rats had 5 x 10(4) adenocarcinoma cells injected into the rectus muscle and blood flow was measured. The experimental group (n = 11) was insufflated with CO(2) at 10 mmHg for 45 min; the control group (n = 11) was not insufflated. After 9 days, tumor weight and volume were analyzed. RESULTS: Clamping caused a 69% reduction of blood flow (p < 0.001), whereas no reduction was registered on the nonclamped side. Tumor weight (p = 0.028) and volume (p = 0.030) were increased on the clamped side. The insufflation of CO(2) caused a 71% reduction of blood flow, whereas no reduction was registered in the control group. Tumor weight (p = 0.006) and volume (p = 0.006) were increased in the insufflated group. CONCLUSION: Clamping, as well as CO(2) insufflation, causes a significant reduction of blood flow in the abdominal wall, which seems to increase tumor growth at the same site.

Abdominal Neoplasms↗
PubMed2006

Efficacy and quality of vessel sealing: comparison of a reusable with a disposable device and effects of clamp surface geometry and structure.

BACKGROUND: During the past few years, a variety of energy-based techniques for vessel ligation have been introduced. With the use of a porcine model and different devices for bipolar vessel sealing (BiClamp and LigaSure), we studied the impact of different clamp surface structures on the efficacy and quality of vessel sealing. METHODS: Eight Swabian Hall pigs underwent splenectomy, nephrectomy, salpingo-oophorectomy, and small bowel resection with the use of bipolar vessel sealing devices designed for open and laparoscopic surgery. Vessel sealing with clamps with a smooth (nonstructured) surface (BiClamp for open surgery and LigaSure for laparoscopic surgery) was compared to that of clamps with a structured (grooved, wafer-like) surface (BiClamp for laparoscopic surgery and LigaSure for open surgery). Measurements of sealed vessels (2- to 7-mm diameter) included the seal failure rate, instrument sticking, and heat-associated morphological vascular wall alterations. RESULTS: Analysis of seal failures did not reveal significant differences between the different devices for both open [BiClamp, 17.9% (17/95); LigaSure, 15.5% (11/71)] and laparoscopic surgery [BiClamp, 2.8% (1/36); LigaSure, 8.6% (3/35)]. Comparing all data of structured versus smooth clamp surfaces, the seal failure rate was lower using clamps with a structured (11.2%) compared to a smooth surface (15.4%). Instrument sticking and thermal spread were found to be significantly increased after sealing with structured surfaces, regardless of whether devices designed for open (p < 0.05 and p < 0.001, respectively) or laparoscopic surgery (p < 0.001 and p < 0.01, respectively) were used. CONCLUSION: Clamps with a structured surface seem to be superior to those with a smooth surface for successful bipolar vessel sealing, as indicated by an increase of thermal spread. However, the more pronounced instrument sticking represents an undesired side effect and should encourage the search for more inert materials to further improve the sealing procedure.

Animals↗
PubMed2004

Pressure-controlled vascular clamp: a novel device for atraumatic vessel occlusion.

Morphological vessel injury after clamping has been an important problem in vascular surgery. Although various techniques have been employed to occlude a vessel during a surgical procedure, a complete, atraumatic occlusion method has not yet been developed. In the present study, we describe use of a novel pressure-controlled and balloon-jawed vessel clamp to provide atraumatic vessel occlusion. We compared its traumatic effect on femoral arteries with that resulting from use of a conventional De Bakey vascular clamp in a canine model. During surgical application, our clamp provided complete vessel occlusion with no oozing in distal arteriotomy. On histological examination of the clamped vessel segments, we observed no destruction of the vessel wall, compared to widespread injury of the De Bakey vascular clamp. Despite some difficulties in application, we think this technique could be a promising method for vascular procedures.

Angioplasty, Balloon, Coronary↗