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PubMed1988

Is the deterioration of liver viability due to hepatic warm ischemia or reinflow of pooled-portal blood in intermittent portal triad cross-clamping?

The effects of hepatic warm ischemia and portal pooling on the viability of the liver were investigated with respect to hepatic energy metabolism by performing intermittent portal triad cross-clamping (Pringle's maneuver) on dogs with or without portosystemic shunt. The dogs were divided into two groups of five: Group 1, non-shunt group, underwent Pringle's maneuver performed for 30 min and declamping for 30 min, a process that was repeated five times; and Group 2, shunt group, underwent the same procedure as Group 1, except for portosystemic shunt using a heparinized hydrophilic catheter between the splenic and jugular veins. The shunt was opened during Pringle's maneuver and was closed immediately at declamping. In the non-shunt group, portal pooling increased and systemic blood pressure decreased when Pringle's maneuver was performed, but in the shunt group portal and systemic blood pressures remained within the normal range. In the non-shunt group, the initial velocity of arterial blood ketone body ratio (KBR) recovery after each declamping significantly (P less than 0.01) decreased from 0.122 +/- 0.016 (per min) after the first declamping to 0.028 +/- 0.017 (per min) after the fifth declamping. Hepatic energy charge [= (ATP + 1/2 ADP)/(ATP + ADP + AMP)] decreased from 0.840 +/- 0.003 before ischemia to 0.749 +/- 0.003 30 min after the fifth declamping (P less than 0.001). The concentrations of lactate and total amino acids in arterial blood increased. On the other hand, in the shunt group, the initial velocity of KBR recovery and hepatic energy charge showed little change even after the fifth declamping (0.081 +/- 0.016 per min and 0.851 +/- 0.009, respectively). The concentrations of lactate and total amino acids showed almost no increase. The impairment of hepatic energy metabolism by intermittent portal triad cross-clamping is mainly due to reinflow of pooled-portal blood to the previously ischemic liver, rather than hepatic warm ischemia. The KBR may be useful for determining the degree of impairment of hepatic energy metabolism.

Amino Acids↗
PubMed1985

The effect of infrarenal aortic clamping and declamping on central haemodynamics and tissue blood flow in pigs--a comparison of fluid replacement between a colloid and a cristalloid.

A study has been undertaken to compare the effect of fluid administration of a cristalloid (Ringer glucose) with that of a colloid (dextran 40) on central haemodynamics and organ blood flow distribution during clamping and after declamping of infrarenal aorta in pigs. The declamping hypotension was significantly greater in the cristalloid group where a decrease in cardiac output was also seen. Tissue blood flow to the kidneys decreased in the cristalloid group during clamping but was maintained in the dextran group. At declamping there was a reactive kidney hyperaemia in the dextran group, while the medullar flow was still significantly decreased in the cristalloid group. The reactive hyperaemia seen in pancreas and small intestines was also greater in the dextran group.

Animals↗
PubMed1980

Membrane current and noise measurements in voltage-clamped node of Ranvier.

In voltage-clamp configurations for nodes of Ranvier the axoplasm resistance functions as a voltage-current converter. In existing configurations this resistance cannot be measured directly. In the present arrangement the electrical resistances of the preparation (axoplasm, membrane and seals) can be measured only from two measurements. This allows us to: 1. calibrate the ionic current under voltage-clamp conditions, and 2. calculate the intensity of the current fluctuations, not arising from the membrane (background noise). The measured axoplasm resistances are considerably higher than the values calculated on the basis of fiber geometry and axoplasm resistivity. The difference is due to the presence of constrictions in the nerve fiber. Membrane current estimation based on geometrical parameters in the presence of wide seals may contain large errors. Variations in the axoplasm resistance for voltage-membrane current conversion were observed within 1.5 hr. In 68% of the fibers this resistance decreased with 30% of the original value. With our current calibration the values for the maximum sodium conductance gNa (at 0mV membrane potential), maximum potassium conductance gK and leakage conductance gL are 49.5 X 10(-8), 6.66 X 10(-8) and 1.71 X 10(-8) S. respectively. The contribution of the different noise sources to the total background noise was calculated at the holding potential. For frequencies below 10(3) Hz there is an excellent agreement between measured and calculated noise levels.

Animals↗
PubMed1983

Changes of ultraviolet sensitivity of voltage-clamped sodium channels during their potential-induced conductance cycle.

The effects of ultraviolet (UV) radiation at various intermediate states of excitation of the frog nerve membrane were measured under voltage-clamp conditions. By synchronizing a UV flash source (flash duration: 10 microseconds) with the stimulating voltage-clamp pulses it was possible to irradiate the membrane at any state of excitation. The resulting UV sensitivities gamma Na and gamma h of gNa and h infinity (V = 0) were compared with those found at the end of a hyperpolarizing prepulse (m = 0, h = 1). The irradiation always induced a decrease of both parameters, gNa and h infinity (V = 0), and the more "h-gates" there were in the closed position at the moment of irradiation, the more pronounced were the effects. The normalized sensitivity of both parameters can be described fairly well by linear relations of the form: gamma x(m,h)/ gamma x(0, 1) = 1 + Px(1 - h) + Qxm2h (x = Na or h). The coefficients P and Q depend on wavelength. The results suggest a conformational change of the nodal membrane during excitation and an interdependence of m and h to some extent.

Animals↗
PubMed1981

Structural responses to voltage-clamping in the toad urinary bladder. II. Granular cells and the natriferic action of vasopressin.

The natriferic action of vasopressin has been investigated with morphological studies of voltage-clamped toad urinary bladders. Granular cell swelling can be induced in the presence of isoosmotic solutions when the orientation of the transmural potential is reversed by voltage clamping (V.A. Bobrycki, J. W. Mills, A.D.C. Macknight & D. R. DiBona, J. Membrane Biol., 60:21, 1981) and results from an increased rate of sodium entry across the mucosal membrane; under these conditions the active transport mechanism at the basal-lateral membrane becomes rate-limiting. Vasopressin exacerbated the voltage-reversal-induced swelling of granular cells while other cell types were unaffected. Granular cell swelling appeared to be dependent upon sodium entry from the mucosal medium since it was completely prevented by amiloride. There was no evidence for an effect of vasopressin on tight junction permeability; voltage-reversal induced the same amount of junction blistering whether or not vasopressin was present. It is concluded that the predominant effect of vasopressin on transepithelial sodium transport is to increase the sodium conductance of the mucosal plasma membrane. As is the case with the hydroosmotic effect of the hormone, the natriferic action of vasopressin seems to be exerted primarily, if not entirely, on the granular cells.

Amiloride↗
PubMed1984

Control of K+ conductance by cholecystokinin and Ca2+ in single pancreatic acinar cells studied by the patch-clamp technique.

The effect of cholecystokinin (CCK) and internal Ca2+ on outward K+ current in isolated pig pancreatic acinar cells has been investigated using the patch-clamp method for whole-cell current recording under voltage-clamp conditions. CCK (2 X 10(-10) M) applied to the bath evoked a marked increase in the outward K+ current associated with depolarizing voltage steps, and this effect was fully reversible and acutely dependent on the presence of external Ca2+. When strongly buffered Ca2+-EGTA solutions were used inside the cells CCK failed to evoke an effect. Increasing the internal Ca2+ concentration [( Ca2+]i) from 5 X 10(-8) M to 10(-7) and 5 X 10(-7) M mimicked the effect of CCK. It would appear therefore that CCK controls K+ conductance in the acinar cells via changes in the internal free ionized Ca2+ concentration.

Animals↗
PubMed1983

Demonstration of vascular redistribution after carotid clamping in rats.

In Long Evans rats, the vertebral arteries supply only the brain stem. Simultaneous application of clamps to both carotid arteries results in a considerable fall of cerebral blood flow and eventually death. If an interval of about 4 days is allowed between the clamping of the 2 carotids, redistribution of blood flow takes place. Owing to this mechanism, 50% of the rats survive with a satisfactory restoration of cerebral blood flow.

Animals↗
PubMed1991

Patch clamping VDAC in liposomes containing whole mitochondrial membranes.

Whole mitochondrial membranes isolated from Neurospora crassa were reconstituted into liposomes and patch clamped. Clear activity characteristic of the mitochondrial channel VDAC was found, namely: open state conductance of 650 pS (in 150 mM KCl, 1 mM CaCl2, 20 mM HEPES, pH 7.2), voltage-dependent closure at both positive and negative potentials, change in conductance upon channel closure of about 450 pS in response to negative and positive potentials, and increased voltage dependence in the presence of König's polyanion. This is the first clear demonstration of VDAC single channels using the patch-clamp technique, even though others used this method before to study whole mitochondrial membranes and liposomes containing mitochondrial proteins. We also found one other channel with a conductance change of about 120 pS.

Electric Conductivity↗
PubMed1990

Supraceliac clamping: an underused alternative in aortic surgery.

In the majority of patients undergoing abdominal aortic surgery, proximal clamping can be readily achieved below the renal vessels. In some situations however, this may be difficult, impossible or ill advised, and an alternative method of control must be sought. We describe the technique and report our personal experience with supraceliac clamping, a maneuver which merits more widespread use in emergency and difficult infrarenal aortic surgery.

Aorta, Abdominal↗
PubMed1991

Pre- and intraoperative transcranial Doppler: prediction and surveillance of tolerance to carotid clamping.

We report 91 patients (mean age 70 years) operated upon, prospectively for a total of 100 carotid revascularizations (nine bilateral). Eighty-five of these patients had pre-, intra-, and postoperative transcranial Doppler investigations. Preoperatively, these 85 patients (92 procedures) were classified into two groups based on the results of their Doppler examinations: Group A (65 patients, 72 procedures), those who did not require an intraoperative indwelling shunt and Group B (20 patients, 20 procedures), those who did. The shunt was inserted only when the mean stump (back) pressure was less than 50 mmHg after cross-clamping. Group A all had satisfactory collaterality with a functional anterior and one or two posterior communicating arteries. Group B had no communicating arteries (anterior or posterior) identified by transcranial Doppler. In 17 of 20 patients in this group, the stump pressure was less than 50 mmHg and a shunt was placed. The overall prediction based on Doppler examination of whether or not patients would need a shunt during operation for the two groups A and B (i.e., 92 procedures) was correct in 95.6% (88/92) of cases. Moreover, six hemodynamically significant stenoses (four in the cavernous portion, two in the middle cerebral artery) were disclosed. Sensitivity and specificity of transcranial Doppler as correlated with arteriographic findings were 70 and 90%. Preoperative transcranial Doppler can measure the velocities of the principal cerebral arteries and the collateral capacity of the circle of Willis, and can forecast tolerance to carotid cross-clamping. Intraoperatively, the velocity of flow in the middle carotid artery was correlated with stump pressure, which allowed for surveillance of the shunt.

Aged↗
PubMed1978

Power density spectra of frog skin potential, current and admittance functions during patch clamp.

Clamp current fluctuations in frog skin of areas down to 0.07 cm2 are dominated by low frequency components (less than 100 Hz). Patch clamp of 0.001 cm2 under high density fluorosilicone oil exhibits components up to 5000 Hz, often including a peak in the current power density spectrum. The admittance spectrum also exhibits a peak at the same frequency. In some skins no peak was observed, but the break in the curve was too sharp to be Lorentzian. In all instances the final limiting slope approached 1/f2. The resonance peak was observed in either Cl- or SO = 4 Ringer's but disappeared when Na+ was replaced with K+. Resonance frequency varied from 100 to 700 Hz.

Animals↗
PubMed1986

In vivo insulin resistance in individual peripheral tissues of the high fat fed rat: assessment by euglycaemic clamp plus deoxyglucose administration.

We have examined peripheral insulin action in conscious rats chronically fed high fat (60% calories as fat) or high carbohydrate (lab chow) diets using the euglycaemic clamp plus 3H-2-deoxyglucose technique. A response parameter of individual tissue glucose metabolic rate (the glucose metabolic index, based on tissue deoxyglucose phosphorylation) was used to assess diet effects in eight skeletal muscle types, heart, lung and white and brown adipose tissue. Comparing high fat with high carbohydrate fed rats, basal glucose metabolism was only mildly reduced in skeletal muscle (only diaphragm was significant, p less than 0.05), but was more substantially reduced in other tissues (e.g. white adipose tissue 61% and heart 33%). No evidence of basal hyperinsulinaemia was found. In contrast, widespread insulin resistance was found during the hyperinsulinaemic clamp (150 mU/l) in high fat fed animals; mean whole body net glucose utilization was 34% lower (p less than 0.01), and the glucose metabolic index was lower in skeletal muscle (14 to 56%, p less than 0.05 in 6 out of 8 muscles), white adipose (27%, p less than 0.05) and brown adipose tissue (76%, p less than 0.01). The glucose metabolic index was also lower at maximal insulin levels in muscle and fat, suggesting the major effect of a high fat diet was a loss of insulin responsiveness. White adipose tissue differed from muscle in that incremental responses (maximal insulin minus basal) were not reduced by high fat feeding. The heart showed an effect opposite to other tissues, with an increase in insulin-stimulated glucose metabolism in high fat versus chow fed rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗
PubMed1984

Prevention of spinal cord injury after cross-clamping of the thoracic aorta.

Paraplegia has been a devastating and unpredictable complication following surgical procedures involving temporary occlusion of the thoracic aorta. This study was undertaken to determine the effect of the pressure gradient between the aortic pressure distal to the occluding aortic clamp and cerebrospinal fluid pressure, defined as "Relative spinal cord perfusion pressure" (RSPP) on the development of the ischemic spinal cord injury. In twelve mongrel dogs, the thoracic aorta just distal to the left subclavian artery was cross-clamped. Somatosensory evoked potentials (SEP) were generated by peripheral stimulation of the bilateral peroneal nerves. After complete loss of SEP was evident, six dogs, Group 1, were subjected to occlusion of the descending thoracic aorta for a period of 20 minutes with maintenance of 0 mmHg of RSPP, by an injection of normal saline into the subarachnoid space. Six other dogs, Group 2, likewise underwent 40 minutes of aortic occlusion, keeping the RSPP at 15 mmHg by withdrawal of cerebrospinal fluid. All the dogs in Group 1 developed paraplegia, whereas all the dogs in Group 2 demonstrated complete postoperative recovery without any neurological sequelae. Thus, RSPP is a most important factor in the development of the ischemic spinal cord injury during the temporary thoracic aortic occlusion.

Animals↗
PubMed1998

Recovery after prolonged cross-clamping tepid blood cardioplegia: report of a case.

Despite a prolonged repetitive aortic cross-clamp time of 411 min, a patient who suffered a left ventricular rupture after undergoing mitral valve replacement following mitral valvuloplasty and Maze procedure recovered without any permanent residual left ventricular dysfunction. During the aortic cross-clamping we used tepid blood cardioplegia for myocardial protection. This case report serves to demonstrate the potential of tepid blood cardioplegia as an effective method of myocardial protection.

Aged↗
PubMed1984

Intravenous nitroglycerin administration during infrarenal aortic clamping.

Twenty-five patients requiring infrarenal abdominal aortic clamping were studied during halothane, nitrous oxide anaesthesia. Aortic clamping caused reductions in cardiac index (CI), stroke volume index (SVI) and left ventricular stroke work index (LVSWI). Systemic vascular resistance (SVR) increased. In seven patients CI was less than 1.81 X min-1 X m-2. Intravenous administration of nitroglycerin, 1 microgram X kg-1 X min-1, for 20 minutes, accompanied by volume loading to maintain pulmonary capillary wedge pressure, resulted in a significant improvement in haemodynamic parameters. CI increased 24 per cent as a result of a 14 per cent increase in SVI and an eight per cent increase in heart rate. LVSWI increased 13 per cent and SVR decreased 21 per cent. The plasma nitroglycerin concentration at the time of these measurements was 2.9 +/- 1.0 ng X ml-1. Aortic unclamping resulted in a mean maximum decrease of 14 +/- 2 torr mean arterial pressure. Epinephrine, norepinephrine and plasma renin activity gradually increased during the period of the anaesthetic.

Aged↗
PubMed1994

[Interactions of intravenous anesthetics with human CNS ion channels. Electrophysiologic studies with a new type of voltage clamp technique].

UNLABELLED: Despite a widespread approach to explain the molecular mechanisms of anaesthetic agents, the complex state of "general anaesthesia" is still not completely understood. Voltage-activated sodium channels from human brain cortex served as a model membrane protein to investigate anaesthetic drug-protein interactions using a novel electrophysiological voltage-clamp technique. Sodium channels are already well-characterized important integral membrane proteins responsible for the generation of the fast-propagated action potential and thus are vital components for neuronal signal integration and cell communication. In order to elucidate the molecular interactions of intravenous anaesthetics with single human brain sodium channels, representative compounds of four different clinical intravenous anaesthetic groups were used to correlate different types of clinical anaesthesia with differential anaesthetic effects on the molecular level. METHODS. Single sodium channels from human brain cortex were incorporated into artificial phospholipid bilayers and studied under our standard experimental conditions (Electrolyte solution: 500 mM NaCl, 10 mM HEPES, pH 7.4, Temp. 22-25 degrees C) with an electrophysiological voltage-clamp technique. In the presence of a channel activator (1 microM batrachotoxin) single-channel characteristics (fractional open time, single-channel conductance and amplitude, steady-state activation behaviour) were characterized for control conditions and in the presence of various doses of four different anaesthetic agents (pentobarbital, propofol, ketamine, midazolam). RESULTS. During control measurements the investigated human brain sodium channels showed stable and reproducible characteristics on the range expected for batrachotoxin-modified sodium channels in bilayers. After completion of the control measurement the effects of the four different general anaesthetics pentobarbital, propofol, ketamine and midazolam were investigated on the same control sodium channels. All four substances demonstrated a blocking effect of sodium channel conductance (pentobarbital: K50: concentration for 50% block of the maximal conductance block: 0.69 mM; blockmax: maximal conductance block (%): 100%; propofol: K50: 0.02 mM, blockmax: 28%; ketamine: K50: 1.1 mM, blockmax: 71%; midazolam: K50: 0.52 mM, blockmax: 100%). Furthermore, a destabilization of the steady-state activation process could be demonstrated. These effects were dose dependent, but only pentobarbital and propofol demonstrated these effects at or near clinically relevant serum concentrations. DISCUSSION: At the clinical level, "general anaesthesia" is a highly complex phenomenon. Similarly, anaesthetics may demonstrate a multimechanistic mode of action also at the molecular level. In this study all four investigated anaesthetic compounds interacted with at least two primary sodium channel functions, leading to a voltage-independent reduction of the fractional channel open time and an interaction with the steady-state activation behaviour, respectively. The effects of pentobarbital and propofol were detectable at concentrations within the range of serum concentrations achieved during clinical anaesthesia, whereas ketamine and midazolam demonstrated qualitatively similar effects exceeding this range 10- to 50-fold. Thus, the human brain sodium channel might serve as a molecular target only for pentobarbital and propofol. This suggests that different types of clinical anaesthesia may correlate with differential actions of anaesthetics on the molecular level.

Brain↗
PubMed2003

An innovative compression-clamp for open contaminated fractures of patella: a short report of five cases.

Five patients with open contaminated fractures of the patella were managed using an indigenously designed external clamp cum compressor. All had an infected lacerated wound at the fracture site, and one had open exposed knee joint (Gustilo grade IIIB open fracture). These patients reported after 3-7 days of injury. The wounds were thoroughly washed and débrided and the clamp cum compressor was applied. They were encouraged to do active knee movements immediately post-operatively. The wounds healed within 3-4 weeks, and the fractures united within 10-16 weeks. Good functional results were achieved in four of the five.

Adult↗