Gene transfer into nerve and muscle by isolated limb perfusion or during replantation

In this study, the authors tested the feasibility of adenovirus vectors transferring functional genetic material into relevent soft-tissue structures during replantation of mouse hindlimbs. An adenovirus vector was constructed encoding the marker gene LacZ and CMV promoter and titered by plaque forming assay to 5 x 10(9) particles/ml. C3H mouse hindlimbs were divided into three groups. In Group 1 (n = 9), the femoral neurovascular bundle was divided and re-anastomosed . Group 2 (n = 9) hindlimbs were transected at mid-femur, perfused with adenovirus, and replanted. Group 3 limbs (n = 4) were perfused with saline only, followed by replantation. After 48 hr, morbidity and mortality were assessed, and the replanted limbs were assayed for gene transfer by histochemistry and polymerase chain reaction. 12/18 limbs were viable after 48 hr. Histochemical staining for adenovirus-mediated LacZ expression was positive within skeletal muscle, femoral nerve, and capillaries adjacent to the anastomoses. Distal muscle was also gene transfer positive. PCR analysis confirmed adenovirus-mediated gene transfer within the femoral nerve and skeletal muscle. This study confirms that viral-mediated gene transfer can be accomplished into the soft tissues of a replanted extremity.     

The changes in the nervous structures under the chemical sympathectomy with guanethidine

The process of sympathectomy of white rats was achieved by injecting guanethidine intraperitoneally every day for 5.5 weeks at doses of 75 mg/kg of animal's weight. This process of sympathectomy was checked according to ganglion cervicothoracicum neurocytes state. The material was taken on days 1, 14, 30, 90 and 180 after cessation of guanethidine injections. It was ascertained that full sympathectomy under prolonged injections of high doses of guanethidine is reached at 5-6 months after the cessation of the injections, though pronounced total degeneration of gnl. stellatum neurocytes is marked on the 14th-30th days after cessation of the guanethidine injections. The following gradient of damaging influence of guanethidine was revealed: sympathetic gnl. cervicothoracicum neurocytes gnl. nodosum afferent neurocytes-neuromuscular synapsis skeletal muscle.     

The sequential metabolic and histological changes occurring in dog leg muscles as a consequence of ischemia (author's transl)

The present study was undertaken to investigate the sequential metabolic and histological changes occurring in dog leg muscles as a consequence of ischemia. The left hind limbs of 45 dogs were amputated at the middle of their thighs and after preserving them in ice water and room temperature for 6 and 12 hours, re-circulation was established. Observation was undertaken on 1) activity of muscle enzymes-CPK, GOT-m, and LDH in the venous blood from the re-circulated limbs, and 2) histological changes of muscles through light microscopy and electron microscopy. Leakage of muscle enzymes into the serum was little in the group preserved in ice water for 6 hours, but resulted in a marked increase in the group preserved in room temperature for 6 hours and 12 hours ischemic groups. This remarkable leakage of muscle enzymes seems to reflect the amount of muscle degeneration. Histologically prominent changes of muscles were characterized by cell swelling and separation of sarcoplasma from endomysium through light microscopic study, and by widening of myofibril space, virtual absence of glycogen granules in sarcoplasma, swelling of mitochondria and appearance of intramitochondrial dense body through electron microscopic study. These histological changes of muscles are well correlated with the leakage of muscle enzymes and demonstrated some extent of irreversible changes had developed already, even in the group preserved in ice water for 6 hours.     

Femoral vein stasis during laparoscopic cholecystectomy: effects of graded elastic compression leg bandages in preventing thrombus formation

Venous stasis of the legs during laparoscopic cholecystectomy was compared between patients without graded compression leg bandages (Group 1; n = 12) and patients with such bandages (Group 2; n = 12) by measuring mean blood flow velocity and cross-sectional area of the femoral vein using a color Doppler ultrasonography. In Group 1, when velocity and area were measured in the supine position, a significant decrease in velocity (p < .05) and a significant increase in area (p < .05) occurred after abdominal insufflation to 10 mm Hg. These changes were greater during abdominal insufflation in the reverse Trendelenburg position than during abdominal insufflation in the supine position. In Group 2, flow velocity was significantly higher (p < .05) before abdominal insufflation as compared with Group 1. After abdominal insufflation to 10 mm Hg and a postural change, velocity significantly decreased (p < .05) and area significantly increased (p < .05) in Group 2, similar to the results in Group 1. During abdominal insufflation at 5 mm Hg or lower, the use of the graded compression bandage was found to be useful for preventing femoral vein stasis. During abdominal insufflation at 10 mm Hg or in the reverse Trendelenburg position, the bandage did not prevent femoral vein stasis.     

Pulse oximetry in the diagnosis of non-critical peripheral vascular insufficiency

Pulse oximetry was used to detect return of pulsatile flow in 27 subjects during reactive hyperaemia following 3 min of total limb ischaemia induced by above knee tourniquet occlusion. Fourteen patients with exercise induced leg pain had 18 symptomatic limbs tested. Thirteen controls had 25 limbs tested. Return of pulsatile flow during reactive hyperaemia occurred within 20 s of tourniquet release in the 25 control limbs which was then regarded as normal. The mean time for return of pulsatile flow in 18 symptomatic limbs was 53 +/- 37 s (P < 0.05 versus controls). Three limbs had a normal value, two of which did not have peripheral vascular disease. Pulse oximetry correctly identified all 25 asymptomatic limbs and 15 of 16 patients with claudication secondary to peripheral vascular disease (PVD). This modification of the reactive hyperaemia test using the pulse oximeter is simple and quick to perform. It has potential as a non-invasive screening test for PVD, suitable for outpatient assessment.     

Nitric oxide synthase is co-localized with vasoactive intestinal polypeptide in postganglionic parasympathetic nerves innervating the rat vas deferens

Cross-sections of the vas deferens taken from control adult male rats showed positive histochemical reactivity to acetylcholinesterase and immunoreactivity for antibodies to protein gene product 9.5, tyrosine hydroxylase, neuropeptide Y, vasoactive intestinal polypeptide, nitric oxide synthase and calcitonin gene-related peptide. Immunoreactivity to substance P was very sparse. Histochemical reactivity to acetylcholinesterase and immunoreactivity to vasoactive intestinal polypeptide and nitric oxide synthase was concentrated in the subepithelial lamina propria and inner smooth muscle layers. Complete surgical denervation resulting from transection of the nerve arising from the pelvic ganglion which supplies the vas deferens totally abolished the immunoreactivity to all of the antibodies tested as well as the histochemical reactivity to acetylcholinesterase. In sections of the prostatic end of the vas deferens taken from rats neonatally pretreated with capsaicin, immunoreactivity to calcitonin gene-related peptide and substance P was reduced by 75 and 83%, respectively. Immunoreactivity to neuropeptide Y, vasoactive intestinal polypeptide and nitric oxide synthase was similar in tissue sections taken from capsaicin-treated rats and those taken from control tissues. Pretreatment of rats with guanethidine or 6-hydroxydopamine decreased immunoreactivity to tyrosine hydroxylase and neuropeptide Y by 60-70%, but immunoreactivity to substance P, vasoactive intestinal polypeptide and nitric oxide synthase was unchanged, while immunoreactivity to calcitonin gene-related peptide and acetylcholinesterase staining was increased by guanethidine but not by 6-hydroxydopamine treatment. Triple labelling experiments showed nitric oxide synthase, vasoactive intestinal polypeptide and acetylcholinesterase all to be co-localized in some nerve fibres. These results indicate that the nitric oxide synthase contained in the nerve fibres innervating the rat vas deferens is unaffected by pretreatment of rats with capsaicin, 6-hydroxydopamine or guanethidine but is abolished by surgical denervation, of postganglionic parasympathetic, sympathetic and sensory nerves. Therefore it appears that nitric oxide synthase is co-localized with vasoactive intestinal polypeptide in the postganglionic parasympathetic nerves which innervate the rat vas deferens.     

The role of focal nerve ischemia and Wallerian degeneration in peripheral nerve injury producing hyperesthesia

BACKGROUND: A new model of pain associated with an experimental peripheral mononeuropathy has stimulated interest in mechanisms of pain and their structural correlates in peripheral nerve, the site of the experimental lesion. METHODS: The pathology of the neuropathy was studied and the results correlated with alterations in nerve blood flow and with the behavioral response to heat applied to the foot. The focal neuropathy was created by loosely tying several ligatures around rat sciatic nerve, which produces hyperesthesia in the ligated limb in 3-5 days. The neuropathology was striking with epineurial and endoneurial vascular stasis, edema, and extensive nerve fiber injury in the ligated segment noted at 1 week after ligation. RESULTS: Nerve blood flow was reduced significantly in the ligated segment during the development of the hyperesthesia response, suggesting that changes in nerve blood flow caused by the ligature compression of the epineurial vessels contributes to the nerve fiber injury and pathophysiology of the model. To further test this hypothesis, the epineurial vasculature was removed from 1-cm lengths of rat sciatic nerve, which reduces nerve blood flow by 58%, and by ligation of the ipsilateral femoral artery, which focally reduces nerve blood flow by 70%, and the behavioral response to heating of the paw was evaluated at 1 week. Crush injury was used as a positive control creating Wallerian degeneration without a substantial reduction in nerve blood flow. CONCLUSIONS: The results suggest that ischemia is an important initial pathogenic mechanism in the hyperesthesia associated with the loose ligature pain model, in so far as it produces Wallerian degeneration and axonal injury. Modest degrees of ischemia producing only demyelination did not produce significant hyperesthesia.     

Chronic thromboembolic pulmonary hypertension--not fully recognized clinical entity

The Xe-133 clearance method is used to measure muscle blood flow in the lower limbs to establish normal values and evaluate the variance between different sex and age groups in healthy Chinese. 99 healthy volunteers were included in this study and divided into three groups according to age and two groups according to sex. Group I: < 40 y/o, 34 persons; Group II: 41-60 y/o, 30 persons; Group III: > 60 y/o, 35 persons; male group: 51 persons; and female group: 48 persons. Intramuscular injection of Xe-133 saline solution in the anterior tibial muscle of the right leg was performed in all the volunteers to calculate muscle blood flow. The muscle blood flow is 3.21 +/- 1.29 ml/100 gm/min in Group I, 3.19 +/- 1.28 in Group II, 2.71 +/- 1.17 in Group III, 3.09 +/- 1.38 in male, and 2.96 +/- 1.24 in female. There was no significant difference in Groups I and II, or in the male and female groups using the Student's t test for unpaired data. However, a mild difference was observed between Groups I and III, and between Groups II and III (P values of 0.09 and 0.11 respectively). We conclude that there is no variance of muscle blood flow in the lower limbs between different sexes and the effect of aging on peripheral muscle blood flow is not significant before 60 in healthy Chinese.     

Motor denervation induces altered muscle fibre type densities and atrophy in a rat model of neuropathic pain

Loose ligation of a sciatic nerve in rats (chronic constriction injury; CCI) provokes sensory, autonomic, and motor disturbances like those observed in humans with partial peripheral nerve injury. So far, it is unknown whether these motor disturbances result from (mechanical) allodynia or from damage to the motor neuron. These considerations prompted us to assess, in CCI rats, the density of motor axons in both the ligated sciatic nerve and the ipsilateral femoral nerve. To this end, we determined the number of cholinesterase positive fibres. It has been demonstrated previously that muscle fibre type density may be used as a measure of motor denervation and/or hypokinesia. Therefore, the myofibrillar ATPase reaction was employed to assess fibre type density in biopsies obtained from the lateral gastrocnemius muscle (innervated by sciatic nerve) and rectus femoris muscle (innervated by femoral nerve). We observed axonal degeneration of motor fibres within the loosely ligated sciatic nerve, both at an intermediate (day 21) and at a late stage (day 90) after nerve injury. The reduction in the number of motor nerve fibres was more pronounced distal to the site of the ligatures than proximal. A (less pronounced) reduction of motor fibres was observed in the ipsilateral (non-ligated) femoral nerve. In line with these findings, we observed altered fibre type densities in muscle tissue innervated by the ligated sciatic nerve as well as the non-ligated femoral nerve indicative of motor denervation rather than hypokinesia. The findings of this study suggest that the motor disorder induced by partial nerve injury involves degeneration of motor nerve fibres not only within the primarily affected nerve but also within adjacent large peripheral nerves. This spread outside the territory of the primarily affected nerve suggests degeneration of motor neurons at the level of the central nervous system.     

Cyclosporine therapy after cardiac transplantation causes hypertension and renal vasoconstriction without sympathetic activation

BACKGROUND: Hypertension frequently complicates the use of cyclosporine A (CyA) therapy, and it has been suggested that sympathoexcitation may be the underlying mechanism in this form of hypertension. METHODS AND RESULTS: To further investigate the possibility of a neurogenic mechanism for this hypertensive effect, we studied the effects of CyA on renal blood flow (n = 11), forearm blood flow (n = 8), and sympathetic nervous system activity, assessed by renal and whole-body radiolabeled norepinephrine plasma kinetics and muscle sympathetic nerve firing (using microneurography) in cardiac transplant recipients receiving CyA and a reference group of healthy age-matched control subjects (n = 17). In 11 cardiac transplant patients (2 hours after cyclosporine dose), renal blood flow was significantly lower than that in 8 control subjects (680 +/- 88 vs 1285 +/- 58 mL/min, P < .001). In 5 of these transplant patients, renal blood flow was measured before and for 2 hours after oral cyclosporine and fell progressively over this period, by 37% (P < .01). Total body and renal norepinephrine spillover rates in transplant patients were similar to those in control subjects (3070 +/- 538 vs 2618 +/- 313 pmol/min and 579 +/- 124 vs 573 +/- 95 pmol/min, respectively), and there was no progressive effect in the 2 hours after cyclosporine dosing. Forearm blood flow was increased 2 hours after CyA administration (1.74 +/- 0.31 to 3.12 +/- 0.50 mL x 100 mL-1 x min-1, P < .001), whereas mean arterial blood pressure and noninvasively determined cardiac output (indirect Fick method) were unchanged. Muscle sympathetic nerve discharge rates recorded in 6 of these transplant patients were not different from those in 9 healthy control subjects (37.9 +/- 10.1 vs 41.3 +/- 2.3 bursts per 100 beats per minute). During 90 to 120 minutes of recording after cyclosporine dosing, nerve firing rates remained unchanged. CONCLUSIONS: CyA therapy causes acute renal vasoconstriction without accompanying systemic hemodynamic effects. These renal effects are nonneural, not being attributable to sympathoexcitation.     

Acute stretching of peripheral nerves inhibits retrograde axonal transport

The conjugation of horseradish peroxidase with wheat germ agglutinin was used to identify the effect on retrograde axonal transport of stretching the rat sciatic nerve indirectly by 10% and 20% femoral lengthening with a unilateral external fixator. To investigate the relationship between retrograde axonal transport and blood flow in the stretched nerve, nerve blood flow in the sciatic nerve was measured by a hydrogen washout technique. At 11% strain (20% femoral lengthening), the numbers of horseradish peroxidase-labelled motor neuron cells and nerve blood flow had decreased by 43% and 50%, respectively. Histological examination demonstrated ischaemic changes, but not mechanical damage. However, at 6% strain (10% femoral lengthening) there were no significant abnormalities. These findings suggest that the inhibition of retrograde axonal transport can be induced by acute stretching of the peripheral nerve and that circulatory disturbance is the main cause of the inhibition of retrograde axonal transport at the low strain.     

Hemodynamic actions of insulin in rat skeletal muscle: evidence for capillary recruitment

Insulin-induced increases in blood flow are hypothesized to enhance overall glucose uptake by skeletal muscle. Whether the insulin-mediated changes in blood flow are associated with altered blood flow distribution and increased capillary recruitment in skeletal muscle is not known. In the present study, the effects of insulin on hemodynamic parameters in rat skeletal muscle in vivo were investigated. Mean arterial blood pressure, heart rate, femoral blood flow, hind leg vascular resistance, and glucose uptake were measured in control and euglycemic insulin-clamped (10 mU x min(-1) x kg[-1]) anesthetized rats. Blood flow distribution within the hind leg muscles was assessed by measuring the metabolism of 1-methylxanthine (1-MX), an exogenously added substrate for capillary xanthine oxidase. Insulin treatment had no effect on heart rate but significantly increased arterial blood pressure (12 mmHg) and femoral blood flow (80%) and decreased hind leg vascular resistance (31%). Changes were similar in magnitude and in time of onset to those reported in humans. Insulin treatment increased hind leg glucose uptake approximately fourfold and also increased hind leg 1-MX metabolism by 50%, suggesting increased exposure to endothelial xanthine oxidase. To ascertain whether the increased 1-MX metabolism was simply due to increased bulk femoral blood flow, epinephrine was infused at a dose (0.125 microg x min(-) x kg[-1]) chosen to match the insulin-induced increase in femoral blood flow. This dose of epinephrine had no significant effects on arterial blood pressure or heart rate but increased femoral blood flow and lowered hind leg vascular resistance to a similar extent as insulin. Epinephrine did not significantly alter 1-MX metabolism as compared with control animals. These results demonstrate that insulin increases total hind leg blood flow and metabolism of 1-MX, suggesting a recruitment of capillary blood flow in rat hind leg not mimicked by epinephrine.     

Implications of the failure of nerve resection and graft to cure chronic pain produced by nerve lesions

Seven patients had developed pain and abnormal sensitivity in the area supplied by a single nerve which had been injured. They were treated unsuccessfully for periods ranging from 3 to 108 months by conservative methods including neurolysis, local anaesthesia, sympathetic blocks, guanethidine, transcutaneous stimulation and analgesics. All then had the damaged nerve resected and in five cases a sural nerve graft was inserted to bridge the resected gap. The patients were then examined 20 to 72 months after the operation. In all seven cases pain and abnormal sensitivity of some intensity recurred in the same area and with the same qualitative characteristic as experienced before the operation. This operation should not be done in patients with this condition. Reasons are given to suggest that peripheral nerve damage induces changes in the central nervous system which are not reversed by treatment directed at the area of the original injury.     

Fulminant Guillain-Barré syndrome with universal inexcitability of peripheral nerves: a clinicopathological study

The pathological basis of nerve inexcitability in Guillain-Barré syndrome has not been established with certainty. We report the clinicopathological findings in a 67-year-old patient with fulminant Guillain-Barré syndrome who died 18 days after onset. Three serial electrophysiological studies revealed nerve inexcitability. Antibodies to Campylobacter jejuni were present but there was no antiganglioside reactivity. Spinal root sections revealed extensive and almost pure macrophage-associated demyelination with occasional presence of T lymphocytes and neutrophil leukocytes. Conversely, in femoral, median, and sural nerves the outstanding lesion was axonal degeneration, with some denuded axons remaining. Unmyelinated fibers, posterior root ganglia, and dorsal columns were preserved. Endoneurial postcapillary venules showed plump endothelial cells with loss of their tight junctions. We conclude that both primary demyelination and axonal degeneration secondary to inflammation account for nerve inexcitability. Our findings lend support to the hypothesis of increased endoneurial pressure as the cause of wallerian degeneration in nerve trunks.     

Mechanisms facilitating oxygen delivery during exercise in patients with chronic heart failure

The aim of the study was to estimate the relative importance of the Bohr effect and redistribution of blood from the non-exercising tissues on the arterial-venous oxygen content differences across the exercising extremities and the central circulation in patients with chronic heart failure; the relationship among femoral vein, systemic and pulmonary artery oxygen partial pressure and hemoglobin saturation was determined. It has been reported that the maximal reduction in femoral vein pO2 precedes peak oxygen consumption and lactic acidosis threshold in patients with chronic heart failure and normal subjects during exercise. The increase in oxygen consumption at work rates above lactic acidosis threshold, therefore, must be accounted for by increase in blood flow in the exercising muscles and right-ward shift on the oxyhemoglobin dissociation curve. Since the total cardiac output increase is blunted in patients with chronic heart failure, diversion of blood flow from non-exercising to exercising tissues may account for some of the increase in muscle blood flow. Ten patients with chronic heart failure performed a progressively increasing leg cycle ergometer exercise test up to maximal effort while measuring ventilation and gas concentration for computation of oxygen uptake and carbon dioxide production, breath-by-breath. Blood samples were obtained, simultaneously, from systemic and pulmonary arteries and femoral vein at rest and every minute during exercise to peak oxygen consumption. At comparable levels of exercise, femoral vein pO2, hemoglobin saturation and oxygen content were lower than in the pulmonary artery. PCO2 and lactate concentration increased steeply in femoral vein and pulmonary artery blood above lactic acidosis threshold (due to lactic acid build-up and buffering), but more steeply in femoral vein blood. These increases allowed femoral vein oxyhemoglobin to dissociate without a further decrease in femoral vein pO2 (Bohr effect). The lowest femoral vein pO2 (16.6 +/- 3.9 mmHg) was measured at 66 +/- 22% of peak VO2 and before the lowest oxyhemoglobin saturation was reached. Artero-venous oxygen content difference was higher in the femoral vein than in the pulmonary artery; this difference became progressively smaller as oxygen consumption increased. "Ideal" oxygen consumption for a given cardiac output (oxygen consumption expected if all body tissues had maximized oxygen extraction) was always higher than the measured oxygen consumption; however the difference between the two was lost at peak exercise. This difference positively correlated with peak oxygen consumption and cardiac output increments at submaximal but not at maximal exercise. In conclusion, femoral vein pO2 reached its lowest value at a level of exercise at or below the lactic acidosis threshold. Further extraction of oxygen above the lactic acidosis threshold was accounted for by a right shift of the oxyhemoglobin dissociation curve. The positive correlation between increments of cardiac output vs "ideal" and measured oxygen consumption suggests a redistribution of blood flow from non-exercising to exercising regions of the body. Furthermore the positive correlation between exercise capacity and the difference between "ideal" and measured oxygen consumption suggests that patients with the poorer function have the greater capability to optimize blood flow redistribution during exercise.     

Leg function after radiotherapy for Ewing's sarcoma

Twenty-nine patients with Ewing's sarcoma of the lower extremity who survived for two or more years following therapy (5000 rad locally and systemic chemotherapy) were studied to assess functional status of the affected leg. Twenty-two of twenty-nine were alive and were reexamined; the deceased patients were evaluated by record review. Twenty-two of the twenty-nine had serial radiographs, which were reviewed to assess growth change induced by radiation. The living patients were divided on the basis of clinical examination into four functional groups with Group I comprising patients with minor functional limitations and leg length discrepancy 1.5 cm or less. Group II patients had moderate functional limitations with 2.5-cm leg-length discrepancy or less. Group III patients had severe functional limitations with up to 4-cm leg length discrepancy. Group IV patients had severe complications, sufficient enough to warrant amputation. Thirteen of twenty-two patients were classified as functional Group I, five as Group II, three as Group III, and one as Group IV. Radiographic changes in growing bone did not correlate with functional results. Although a femoral fracture and an age less than 16 years at diagnosis were found to be less favorable prognostic factors for the functional treatment result, these results show that neither femoral location nor young age justify primary amputation for Ewing's sarcoma of the lower leg extremity.     

Release of somatostatin and its role in the mediation of the anti-inflammatory effect induced by antidromic stimulation of sensory fibres of rat sciatic nerve

1. The effect of antidromic stimulation of the sensory fibres of the sciatic nerve on inflammatory plasma extravasation in various tissues and on cutaneous vasodilatation elicited in distant parts of the body was investigated in rats pretreated with guanethidine (8 mg kg(-1), i.p.) and pipecuronium (200 microg kg(-1), i.v.). 2. Antidromic sciatic nerve stimulation with C-fibre strength (20 V, 0.5 ms) at 5 Hz for 5 min elicited neurogenic inflammation in the innervated area and inhibited by 50.3 +/- 4.67% the development of a subsequent plasma extravasation in response to similar stimulation of the contralateral sciatic nerve. Stimulation at 0.5 Hz for 1 h also evoked local plasma extravasation and inhibited the carrageenin-induced (1%, 100 microl s.c.) cutaneous inflammation by 38.5 +/- 10.0% in the contralateral paw. Excitation at 0.1 Hz for 4 h elicited no local plasma extravasation in the stimulated hindleg but still reduced the carrageenin-induced oedema by 52.1 +/- 9.7% in the paw on the contralateral side. 3. Plasma extravasation in the knee joint in response to carrageenin (2%, 200 microl intra-articular injection) was diminished by 46.1 +/- 12.69% and 40.9 +/- 4.93% when the sciatic nerve was stimulated in the contralateral leg at 0.5 Hz for 1 h or 0.1 Hz for 4 h, respectively. 4. Stimulation of the peripheral stump of the left vagal nerve (20 V, 1 ms, 8 Hz, 10 min) elicited plasma extravasation in the trachea, oesophagus and mediastinal connective tissue in rats pretreated with atropine (2 mg kg(-1), i.v.), guanethidine (8 mg kg(-1), i.p.) and pipecuronium (200 microg kg(-1), i.v.). These responses were inhibited by 37.8 +/- 5.1%, 49.7 +/- 9.9% and 37.6 +/- 4.2%, respectively by antidromic sciatic nerve excitation (5 Hz, 5 min) applied 5 min earlier. 5. Pretreatment with polyclonal somatostatin antiserum (0.5 ml/rat, i.v.) or the selective somatostatin depleting agent cysteamine (280 mg kg(-1), s.c.) prevented the anti-inflammatory effect of sciatic nerve stimulation (5 Hz, 5 min) on a subsequent neurogenic plasma extravasation of the contralateral paw skin. The inhibitory effect of antidromic sciatic nerve excitation on plasma extravasation in response to vagal nerve stimulation was also prevented by somatostatin antiserum pretreatment. 6. Cutaneous blood flow assessment by laser Doppler flowmetry indicated that antidromic vasodilatation induced by sciatic nerve stimulation was not inhibited by excitation of the sciatic nerve of the contralateral leg (1 Hz, 30 min) or by somatostatin (10 microg/rat, i.v.) injection. 7. Plasma levels of somatostatin increased more than 4 fold after stimulation of both sciatic nerves (5 Hz, 5 min) but the stimulus-evoked increase was not observed in cysteamine (280 mg kg(-1), s.c.) pretreated rats. 8. These results suggest that somatostatin released from the activated sensory nerve terminals mediates the systemic anti-inflammatory effect evoked by stimulating the peripheral stump of the sciatic nerve.     

Comparison of nerves to cerebral and extracerebral blood vessels: a differential effect of alpha methyl tyrosine on norepinephrine content

Alpha methyl tryosine (AMT), and inhibitor of norepinephrine (NOR) synthesis, was injected intraperitoneally (200 mg/kg) in Sprague Dawley rats, kept in a cold room, or at room temperature for 16 hours. Using formaldehyde induced NOR fluorescence, nerve counts were made on whole mounts of cerebral and femoral arterioles 14-300 micronm in diameter, utilizing a grid superimposed on the vessels. Cold had no effect on the number of visible (i.e. fluorescing) nerves. AMT had an appreciable effect but only on nerves to femoral arterioles, where a significant reduction in nerve count was observed in both cold stressed and non stressed rates, when compared with animals not given AMT. Since the counting technique is sensitive only to large depletions of NOR, we cannot conclude that AMT failed to affect NOR content in cerebrovascular nerves. However, if such an effect was present, it was much less than the effect of AMT on nerves to femoral vessels. We suggest that the differential effect of AMT on these 2 vascular beds may indicate a lower basal level of NOR release from cerebrovascular nerves, which would correlate with the difficulty of demonstrating basal sympathetic tone in this vascular bed.     

Local reflex in microcirculation in human cutaneous tissue

Blood flow in cutaneous tissue measured by the local 133Xenon washout technique decreased about 35 per cent during venous stasis of 40 mmHg in three normal subjects. The response was unaffected by block of the nerve three cm proximally to the labeled area. When the tissue was infiltrated with lidocaine or with phentolamine, blood flow remained constant, indicating that the decrease in blood flow is due to an arteriolar vasoconstrictor response to increase in venous transmural pressure. Local venous stasis elicited a vasoconstrictor response in an adjoining area not affected by the stasis. The response was blocked by lidocaine applied to the side of stasis. In 2 chronically sympathectomized patients, the vasoconstrictor response was abolished in the denervated limbs but present on the non-operated side (1 patient with unilateral sympathectomy). The results indicate that the vasoconstrictor response to an increase in venous transmural pressure is due to a local nervous mechanism involving sympathetic adrenergic fibres, most likely a sympathetic axon reflex.     

Adrenergic neuronal degeneration induced in portal vein and caudal artery by 6-hydroxydopamine in vitro

Fluorescence histochemistry and electron microscopy were used to study the structural consequences of in vitro exposure to the sympatholytic agent 6-hydroxydopamine on two blood vessels, the portal mesenteric vein and caudal artery of the rat. The results showed depletion of catecholamines to indetectable levels associated with clear signs of adrenergic nerve degeneration, such as cytoplasmic shrinking, virtual absence of dense core vesicles and swelling of mitochondria. All of the changes observed occurred within 2 hours in the caudal arteries and 3.5 hours in the portal veins. Comparison of electron and fluorescence micrographs of incubated control specimens with those of unincubated, fresh specimens showed that the nerve endings of the incubated controls were well preserved for at least 3.5 hours. With destruction of nerve endings in such a short period of time, the processes of specific neuronal degeneration could be clearly demonstrated in isolated blood vessels.