Venous Effluent Blood Research Articles

PD-0020 Pharmacokinetic Analysis of Percutaneous Hepatic Perfusion (PHP) of Melphalan in Patients with Hepatic Metastases from Melanoma

ABSTRACT Introduction PHP (Delcath Systems Inc., New York, NY) is a non-invasive regional therapy that isolates the liver using a series of catheters, allowing infusion of high doses of chemotherapy directly into the hepatic artery. Systemic toxicity is minimized by extracorporeal hemofiltration of effluent hepatic venous blood, which is then returned to the systemic circulation. A randomized phase III study was conducted to compare PHP of high-dose melphalan with best alternative care in patients with ocular or cutaneous melanoma metastatic to the liver. A pharmacokinetic analysis, including an evaluation of filter extraction efficiency, was performed in a subset of patients from this study. Methods Melphalan was perfused into the liver over 30 minutes via PHP at a maximum dose of 3.0 mg/kg of ideal body weight (maximum 220 mg/treatment). Extracorporeal hemofiltration was performed during and for 30 minutes after melphalan infusion. PHP was performed with patients under general anesthesia. Blood samples (7 mL) were collected during the first cycle of PHP melphalan simultaneously from the periphery (arterial line) and extracorporeal circuit (both pre- and post-filter). Sample collections were made at: baseline (peripheral only); 15 minutes after the start of the infusion; immediately post-infusion; 5, 10, 15, and 30 minutes post-infusion. Plasma concentrations of melphalan were determined by high-pressure liquid chromatography with ultraviolet detection. Data were analyzed using a non-compartmental approach with WinNonlin v5.2 (Pharsight Corporation, Mountain View, CA). Concentration-time profiles were constructed for each sampling location (i.e. three profiles/patient). Pharmacokinetic parameters were: maximum plasma concentration (Cmax); area under the concentration-time curve from time zero to final sample (AUClast) calculated using the linear trapezoidal method; and filter efficiency [defined as: (pre-filter AUClast) minus (post-filter AUClast) divided by (pre-filter AUClast)]. Download : Download full-size image Figure . Individual AUC by site, alter 3 0 mg/Kg (ideal body weight) melphalan Horizontal bars represent mean and 95% Cls Results Plasma samples were available from 48 patients, of which 40 were evaluable. Mean absolute melphalan dose was 191 mg (range, 137–220 mg) and duration of perfusion was 30 minutes (range, 16–52 minutes). Mean prefilter, postfilter and systemic Cmax values were 8726 ng/mL, 2330 ng/mL and 1429 ng/mL, respectively. Mean prefilter AUClast was 264,652 min*ng/mL, mean postfilter AUClast was 74,146 min*ng/mL and mean systemic AUClast was 50,777 min*ng/mL; individual patient data for AUClast are shown in the Figure. Mean filter extraction efficiency was 71.2% ± 10.4%. Filter efficiency did not change significantly with absolute melphalan dose (p = 0.86, Spearman) or theoretical rate of perfusion (p = 0.064, Spearman). Conclusion The mean filter extraction efficiency of the PHP filtration system is 71%, and results appear to be consistent across patients (narrow 95% CI intervals). These findings indicate that the filter removes most of the melphalan administered via PHP.

Enterocyte Shedding and Epithelial Lining Repair Following Ischemia of the Human Small Intestine Attenuate Inflammation

BackgroundRecently, we observed that small-intestinal ischemia and reperfusion was found to entail a rapid loss of apoptotic and necrotic cells. This study was conducted to investigate whether the observed shedding of ischemically damaged epithelial cells affects IR induced inflammation in the human small gut.Methods and FindingsUsing a newly developed IR model of the human small intestine, the inflammatory response was studied on cellular, protein and mRNA level. Thirty patients were consecutively included. Part of the jejunum was subjected to 30 minutes of ischemia and variable reperfusion periods (mean reperfusion time 120 (±11) minutes). Ethical approval and informed consent were obtained. Increased plasma intestinal fatty acid binding protein (I-FABP) levels indicated loss in epithelial cell integrity in response to ischemia and reperfusion (p<0.001 vs healthy). HIF-1α gene expression doubled (p = 0.02) and C3 gene expression increased 4-fold (p = 0.01) over the course of IR. Gut barrier failure, assessed as LPS concentration in small bowel venous effluent blood, was not observed (p = 0.18). Additionally, mRNA expression of HO-1, IL-6, IL-8 did not alter. No increased expression of endothelial adhesion molecules, TNFα release, increased numbers of inflammatory cells (p = 0.71) or complement activation, assessed as activated C3 (p = 0.14), were detected in the reperfused tissue.ConclusionsIn the human small intestine, thirty minutes of ischemia followed by up to 4 hours of reperfusion, does not seem to lead to an explicit inflammatory response. This may be explained by a unique mechanism of shedding of damaged enterocytes, reported for the first time by our group.

Determination of carcinoembryonic antigen levels in peripheral and draining venous blood in patients with colorectal carcinoma

The problem of the relationship between blood carcinoembryonic antigen (CEA) levels and tissue CEA content in colorectal carcinoma, and the mechanisms for CEA release from tumor cells in tissue adjacent to the neoplasm is important to understanding the biology of colorectal carcinoma. It has not been adequately explained whether CEA in the peripheral blood is drained mainly by portal system blood or by the lymphatic system, or indeed by both systems. To study the behavior of CEA levels in peripheral blood (CEA-p) and venous effluent blood (CEA-d) among patients with colorectal tumors, who underwent curative operation. A total of 28 patients were studied (12 male [42.9%] and 16 female [57.1%], mean age 66.1 years [range: 43 - 84]). Immediately after laparotomy, peripheral venous blood was extracted by antecubital venous puncture and venous effluent blood was collected from the main drainage vein of the lesions. Values of CEA-p, CEA-d and the gradient between CEA-d and CEA-p that were less than 5.0 ng/mL were considered normal. Eight (28.6%) patients were stage A in Duke's classification, nine (32.1%) stage B and 11 (39.3%) stage C. The neoplasm was located in the rectum of 14 patients (50.0%), in the transverse colon in five (17.9%), in the sigmoid in four (14.3%), in the cecum and/or ascending colon in three (10.7%), and in the descending colon in two (7.1%). The histopathological examination revealed well-differentiated adenocarcinoma in all the patients. Only one patient (3.6%), Duke's classification stage C, presented neoplasm with venous invasion. The gradient between the CEA-p and CEA-d levels were normal in 25 patients (88.3%) and high in three (10.7%). The mean value for CEA-p was 3.8 +/- 4.1 ng/mL (0.1-21.1 ng/mL) and for the drained CEA (CEA-d) it was 4.5 +/- 4.3 ng/mL (0.3-20.2 ng/mL), without significant difference between these values. There was a significant difference between the mean value for CEA-p and CEA-d levels greater than 5 ng/mL. The CEA-p and CEA-d levels in the colorectal carcinoma patients were not shown to be different. The results from this study suggest that, in colorectal neoplasm without venous invasion, there may not be notable CEA drainage from the tumor by the portal vein effluent blood.

Time course of superoxide generation in reperfusion after arterial ischaemia and venous congestion

In order to identify differences in superoxide (O2-) production after reperfusion following arterial ischaemia and venous congestion, we directly quantified O2- in the venous effluent from reperfused hindlimbs in the rabbit. Using a total of 20 rabbit hindlimbs, we made hindlimb-reperfusion models by section of both soft tissue and bony structures exclusive of femoral vessels around the thighs and clamping of the artery alone (group A: n = 10) or vein alone (group V: n = 10) by microvascular clamp. A pilot study revealed that the critical time for venous congestion was 2 h. Accordingly, clamping was performed at 25 degrees C (normothermia) for 2 h. Sham operations were performed in the control (n = 7). Venous effluent blood samples from the femoral vein proximal to the clamping point were collected before reperfusion, and from 5 to 60 min after reperfusion at 5 min intervals. Effluent blood samples were used to quantify O2- using a chemiluminescence method using a derivative of luciferin. The time course of O2- production after reperfusion in group A and group V showed significant differences (P < 0.05), compared with the controls. There was no significant difference in the time course of O2- production in group A and that in group V. In the early reperfusion phase, no difference in O2- production was observed after arterial ischaemia or venous congestion. In future studies, we will test the contributions of free radicals by setting longer ischaemic times, longer reperfusion times, and studying secondary ischaemia.

Stimulatory effect of insulin on calcitonin secretion.

Infusion of insulin directly into thyroid arterial blood perfusing the surgically isolated in situ pig thyroid gland produced an increase in the secretion rate of calcitonin (CT) measured by immunoassay in thyroid venous effluent blood. Insulin in concentrations ranging from approximately 1 to 400 ng/ml produced a maximal stimulation of 4-5 fold. The stimulatory effect of insulin on CT could not be duplicated by infusion of either IGF-I or amylin. Specific binding of radiolabeled insulin was demonstrated using isolated pig thyroid plasma membranes and both rat (6-23) and human (TT) medullary thyroid carcinoma C-cells. Increased CT release was observed from C-cells exposed to a high concentration of insulin. The administration of glucose iv to pigs in order to stimulate secretion of endogenous insulin produced an increase in circulating insulin, which was accompanied by an increase in the secretion of CT. The results show that insulin, delivered directly to the pig thyroid gland, can stimulate CT release. The in vitro binding and secretion studies indicate that C-cells can bind insulin and respond with an increase in CT secretion, and the iv glucose experiments suggest that endogenous insulin is capable of stimulating CT secretion. The findings imply that insulin is capable of acting as a CT secretagogue and suggest that changes in CT secretion may accompany altered states of insulin production such as diabetes or insulin-secreting tumors.

Relationship between flushing pressure and nitric oxide production in preserved lungs.

Nitric oxide (NO) is considered to be one of the endogenous inhibitory factors of ischemic reperfusion injury. In this study, the NO-producing ability of the preserved lung, flushed at various pulmonary artery pressures (flushing pressure), was studied during reperfusion using an ex vivo rabbit lung perfusion model. The lungs were flushed with 200 ml of preservation solution with flushing pressures adjusted to 15, 15, 20, and 25 mmHg for groups 1, 2, 3, and 4, respectively (n=5 in each group). In the control group (group 1), the heart-lung block was harvested after flushing and the lungs were assessed without preservation. In the other groups, the harvested blocks were preserved at 8 degrees C for 24 hr and reperfused with homologous blood for pulmonary functional assessment. Pulmonary function was assessed by measuring mean airway pressure, mean pulmonary arterial pressure, partial oxygen tension of pulmonary venous effluent blood, and pulmonary wet-dry weight ratio. The sequential changes in the concentration of NO-related substances (NO-RS) in the serum of reperfused blood were also measured by chemiluminescence. During reperfusion, biphasic increases in NO-RS were observed in all groups. In groups 3 and 4, the increases in NO-RS were significantly lower than those of groups 1 and 2, and pulmonary function deteriorated. These data suggest that in order to maintain the endogenous NO-producing ability of preserved lung, the flushing pressure must be less than 20 mmHg.

Roles of Glucose Transport and Glucose Phosphorylation in Muscle Insulin Resistance of NIDDM

Insulin resistance for glucose metabolism in skeletal muscle is a key feature in NIDDM. The quantitative role of the cellular effectors of glucose metabolism in determining this insulin resistance is still imperfectly known. We assessed transmembrane glucose transport and intracellular glucose phosphorylation in vivo in skeletal muscle in nonobese NIDDM patients. We performed euglycemic insulin clamp studies in combination with the forearm balance technique (brachial artery and deep forearm vein catheterization) in five nonobese NIDDM patients and seven age- and weight-matched control subjects (study 1). D-Mannitol (a nontransportable molecule), 3-O-[14C]methyl-D-glucose (transportable, but not metabolizable) and D[3-3H]glucose (transportable and metabolizable) were simultaneously injected into the brachial artery, and the washout curves were measured in the deep venous effluent blood. In vivo rates of transmembrane transport and intracellular phosphorylation of D-glucose in forearm muscle were determined by analyzing the washout curves with the aid of a multicompartmental model of glucose kinetics in forearm tissues. At similar steady-state concentrations of plasma insulin (approximately 500 pmol/l) and glucose (approximately 5.0 mmol/l), the rates of transmembrane influx (34.3 +/- 9.1 vs. 58.5 +/- 6.5 micromol x min(-1) x kg(-1), P < 0.05) and intracellular phosphorylation (5.4 +/- 1.6 vs. 38.8 +/- 5.1 micromol x min(-1) x kg(-1), P < 0.01) in skeletal muscle were markedly lower in the NIDDM patients than in the control subjects. In the NIDDM patients (study 2), the insulin clamp was repeated at hyperglycemia, (approximately 13 mmol/l) trying to match the rates of transmembrane glucose influx measured during the clamp in the controls. The rate of transmembrane glucose influx (62 +/- 15 micromol x min(-1) x kg(-1)) in the NIDDM patients was similar to the control subjects, but the rate of intracellular glucose phosphorylation (16.6 +/- 7.5 micromol x min(-1) x kg(-1)), although threefold higher than in the patients during study 1 (P < 0.05), was still approximately 60% lower than in the control subjects (P < 0.05). These data suggest that when assessed in vivo, both transmembrane transport and intracellular phosphorylation of glucose are refractory to insulin action and add to each other in determining insulin resistance in skeletal muscle of NIDDM patients. It will be of interest to compare the present results with the in vivo quantitation of the initial rate of muscle glucose transport when methodology to perform this measurement becomes available.

Evidence for free radical generation after primary percutaneous transluminal coronary angioplasty recanalization in acute myocardial infarction

In animal models, oxygen-derived free radicals have been found to be important mediators of reperfusion injury to ischemic but viable myocardium. However, in humans, there is no direct evidence of free radical production after the restoration of coronary artery patency in acute myocardial infarction. The purpose of this study was to quantitate and assess the time course of free radical production in coronary venous outflow in patients with acute myocardial infarction undergoing successful recanalization of the infarct-related artery by primary percutaneous transluminal coronary angioplasty (PTCA). Primary PTCA was performed in 17 patients with acute myocardial infarction of <6 hours duration. Direct free radical production was assessed by coronary venous effluent blood sampling before PTCA and at timed intervals up to 24 hours (or 48 hours in 6 patients) after recanalization. All samples were added to the spin trapping agent α-phenyl N-tert butyl nitrone and analyzed by electron paramagnetic resonance spectroscopy. Vessel patency resulted in a sharp increase in free radical signal. Relative to the level before PTCA, the changes reached statistical significance after only 15 minutes (p <0.05). Peak signals were observed between 1 1 2 and 3 1 2 hours (p < 0.001), then declined up to 5 hours. A second increase in signal level was detected between 18 and 24 hours despite no angiographic evidence of reocclusion. A gradual decline was observed after 24 hours. These findings provide the first direct and quantitative evidence of free radical production in the immediate postrecanalization phase after thrombotic occlusion of a major coronary artery in humans.

Induction of tumour cell shedding into effluent venous blood breast cancer surgery

Surgeons have long been concerned that cancer may be disseminated by shedding of tumour cells into the bloodstream during surgery. Early claims that cancer operations induced an increase in the number of tumour cells shed into the circulation were subsequently discredited, and the issue has remained unresolved. We used immunocytochemistry for cytokeratins to detect tumour cells in effluent blood from breast carcinomas in 18 patients undergoing surgery. Tumour cells were detectable in 6/18 patients during surgery, in only one patient before operation and in none post-operatively (P = 0.025). Circulating cells were associated with vascular invasion within the primary tumour (P = 0.032). No cytokeratin-positive cells were found in blood from ten normal volunteers or four patients undergoing surgery for other breast conditions. These results confirm that cancer surgery in humans results in an increase in the shedding of tumour cells into the circulation. The implications for prognosis and practice should be determined by larger prospective studies.

Purine metabolite washout and platelet aggregation at reflow after tourniquet ischemia: effect of intravenous regional lidocaine.

Tourniquet ischemia will influence the biochemical milieu of tissue cells and affect the metabolism of purines in skeletal muscle distal to the occlusion. At reperfusion, generation of oxygen radicals by the hypoxanthine-xanthine oxidase system may ensue, influencing white blood cell and thrombocyte aggregation, causing damage to the endothelial cell barrier and inducing non-reflow type phenomena. Amide-type local anaesthetics are known to affect local vasotone, leukocyte adherence and platelet function but the influence of lidocaine on purine metabolite washout and platelet aggregation following tourniquet ischemia for lower limb surgery is not known in detail. Therefore, the effects of regional intravenous lidocaine during tourniquet ischemia for knee surgery on purine catabolite washout and platelet function following reflow were assessed. Eight patients served as control (C-group) and 8 (L-group) received 100 ml of lidocaine (2.5 mg/ml) in the emptied venous bed of the leg to be operated. All patients had spinal anaesthesia (0.5% bupivacaine). Effluent venous blood from the leg and radial arterial blood was collected and analysed for xanthine (X), hypoxanthine (HX), base excess (BE), pH and potassium before and after reperfusion. Platelet ADP-induced aggregation (ADP-agg.) and secretion of beta-thromboglobulin (beta-TG) were measured in the effluent blood as well as systemically. After tourniquet release (TR), X and HX were significantly increased in effluent venous blood but the washout was enhanced in the L-group during the initial reperfusion phase. BE was significantly higher in the L-group both before and after TR whereas pH and potassium washout was comparable between the groups. No systemic effects on platelets were detected after tourniquet release but ADP-agg. in effluent venous blood was attenuated in 6 out of 8 patients in the L-group (NS). It is concluded that HX and X are generated during leg ischemia. Regional intravenous lidocaine, most probably through a vasodilatory mechanism and inhibition of white blood cell activation, may attenuate non-reflow phenomena and thereby exert beneficial effects on post-ischemic recovery by enhancing post-ischemic tissue reperfusion.

Association between tumour angiogenesis and tumour cell shedding into effluent venous blood during breast cancer surgery.

Tumour angiogenesis is a powerful prognostic indicator in breast cancer. Shedding of tumour cells into the bloodstream is essential for haematogenous metastasis. The relation between cell shedding and angiogenesis in human cancer is not known. We studied vascular density and cell shedding in 16 women undergoing breast cancer surgery. Circulating cells were found in one patient before, in six during, and none after operation. Vascular density was related to detection of circulating tumour cells during surgery (correlation coefficient for first intraoperative sampling = 0.56, p = 0.024, 95% CI 0.08 to 0.83). Tumour cell shedding during surgery is related to vascular density; the relevance of this relation to the prognostic significance of vascular density in breast cancer merits further study.

Transmembrane glucose transport in skeletal muscle of patients with non-insulin-dependent diabetes.

Insulin resistance for glucose metabolism in skeletal muscle is a key feature in non-insulin-dependent diabetes mellitus (NIDDM). Which cellular effectors of glucose metabolism are involved is still unknown. We investigated whether transmembrane glucose transport in vivo is impaired in skeletal muscle in nonobese NIDDM patients. We performed euglycemic insulin clamp studies in combination with the forearm balance technique (brachial artery and deep forearm vein catheterization) in six nonobese NIDDM patients and five age- and weight-matched controls. Unlabeled D-mannitol (a nontransportable molecule) and radioactive 3-O-methyl-D-glucose (the reference molecular probe to assess glucose transport activity) were simultaneously injected into the brachial artery, and the washout curves were measured in the deep venous effluent blood. In vivo transmembrane transport of 3-O-methyl-D-glucose in forearm muscle was determined by computerized analysis of the washout curves. At similar steady-state plasma concentrations of insulin (approximately 500 pmol/liter) and glucose (approximately 5.15 mmol/liter), transmembrane inward transport of 3-O-methyl-D-glucose in skeletal muscle was markedly reduced in the NIDDM patients (6.5 x 10(-2) +/- 0.56 x 10(-2).min-1) compared with controls (12.5 x 10(-2) +/- 1.5 x 10(-2).min-1, P < 0.005). Mean glucose uptake was also reduced in the diabetics both at the whole body level (9.25 +/- 1.84 vs. 28.3 +/- 2.44 mumol/min per kg, P < 0.02) and in the forearm tissues (5.84 +/- 1.51 vs. 37.5 +/- 7.95 mumol/min per kg, P < 0.02). When the latter rates were extrapolated to the whole body level, skeletal muscle accounted for approximately 80% of the defect in insulin action seen in NIDDM patients. We conclude that transmembrane glucose transport, when assessed in vivo in skeletal muscle, is insensitive to insulin in nonobese NIDDM patients, and plays a major role in determining whole body insulin resistance.

Role of potassium in the reflex regulation of blood pressure during static exercise in man.

1. The relationship between [K+] in venous effluent blood and alterations in mean arterial blood pressure was studied during static handgrip contractions at 15 and 30% of maximal voluntary contraction (MVC). 2. To further elucidate the importance of K+ in the reflex regulation of blood pressure a situation with normal recovery was compared with a situation in which 3 min of post-exercise occlusion was applied by arresting the circulation to the forearm just prior to the cessation of the contraction. 3. There was a temporal as well as quantitative correlation between venous [K+] and the blood pressure response during and after static exercise. During 30% MVC mean arterial blood pressure (MAP) attained 161.7 mmHg and venous [K+] 5.8 mM, while the corresponding values during 15% MVC were 121.5 mmHg and 5.0 mM. 4. In the occlusion period mean arterial blood pressure remained elevated above resting level and provided a measure of the magnitude of muscle chemoreflexes. In the same period venous [K+] was maintained at 5.3 mM and 4.6 mM following 30% MVC and 15% MVC respectively. This is indicative of interstitial concentrations of above 8-10 mM. This level is sufficiently high to stimulate type III and IV muscle afferents involved in the reflex regulation of blood pressure, and strengthens the notion that K+ may play an important role in eliciting the pressor reflex. 5. In contrast to [K+] the time course of venous blood concentrations of lactate and ammonia (NH3) exhibited a clear dissociation from the blood pressure recordings.

Atropine‐resistant submandibular responses to stimulation of the parasympathetic innervation in the anaesthetized ferret.

1. Submandibular salivary and vascular responses to stimulation of the peripheral end of the chorda-lingual nerve at 20 Hz continuously for 60 min were investigated in anaesthetized ferrets, in which the sympathetic innervation to the gland was cut, in the presence and absence of atropine (2.0 mg kg-1). 2. Both the increase in submandibular salivary flow and protein output, which occurred in response to nerve stimulation, were substantially reduced following the administration of atropine, the latency was greatly increased thereby, and both responses were more transient but neither was abolished by atropine. The fall in submandibular vascular resistance was not significantly affected by atropine, either in respect of extent or duration. 3. Chorda-lingual stimulation produced an increase in the output of vasoactive intestinal peptide (VIP), substance P (SP) and calcitonin gene-related peptide (CGRP) in the submandibular venous effluent blood. Each of these responses was maximal within the first 10 min after the onset of stimulation and declined thereafter. The time-scales of both the CGRP and SP responses were similar to those of the atropine-resistant secretory responses, both being quite short-lived, whereas the output of VIP (like the atropine-resistant vascular response) was significantly greater than the basal value throughout the whole of the 60 min period of stimulation. 4. The CGRP response was completely abolished by pre-treatment with atropine, whereas the outputs of both VIP and SP were significantly enhanced thereby. Both the submandibular vascular and secretory responses to chorda-lingual stimulation were almost completely suppressed following the administration of hexamethonium, and there was then no detectable release of peptidergic agonists from the gland. 5. The atropine-resistant submandibular salivary secretory responses were completely abolished by pre-treatment with a tachykinin inhibitor [( D-Arg1, D-Cl2 Phe5, Asn6, D-Trp7,9, Nle11]-SP; 0.75 mg kg-1) without affecting the fall in submandibular vascular resistance. 6. Following pre-treatment with hexamethonium, I.V. bolus injections of methacholine, SP and CGRP elicited increases in submandibular blood flow and secretion of saliva. VIP caused an increase in blood flow without overt secretion, although it is known to increase secretion of protein and to potentiate the secretory response to SP. Taken together, all these results are consistent with the contention that VIP contributes to the vasodilator response to stimulation of the para-sympathetic innervation in this gland and that both SP and CGRP are likely to contribute to the secretory response.

5‐Hydroxytryptamine and arachidonic acid metabolites modulate extensive platelet activation induced by collagen in cats in vivo

1. The pathways contributing to the platelet adhesion/aggregation reaction elicited by collagen microfibrils, administered to cats in vivo, were analysed. 2. The intra-aortic infusion of collagen (100 micrograms kg-1 in 1 min) caused an extensive activation of platelets, as evidenced by the time-dependent drop of free platelet numbers in whole blood, and the increases of 5-hydroxyindoles (5-HI), 5-hydroxytryptamine (5-HT) and thromboxane B2 (TXB2) levels in plasma, prepared from effluent venous blood sampled from the inferior caval vein. 3. 5-HT2 receptor blockade with ketanserin (0.63 mg kg-1 i.v., 10 min) and cyclo-oxygenase inhibition with aspirin (10 mg kg-1 i.v., 10 min) slightly attenuated the peak reduction of free platelets in whole blood in response to collagen without affecting changes in plasma 5-HI. Aspirin, but not ketanserin, reduced the collagen-induced changes in plasma TXB2, prostaglandin E2 (PGE2) and 6K-PGF1 alpha. 4. Dual TXA2 synthetase inhibition/TXA2-prostaglandin endoperoxide receptor antagonism with ridogrel (5 mg kg-1 i.v., 10 min) halved the drop in free platelets, reduced the release of platelet 5-HI, inhibited the increase in plasma TXB2 and elevated that of 6K-PGF1 alpha and PGE2 in response to collagen. 5. Combined treatment with ketanserin and aspirin reduced the collagen-induced drop of free platelets and the release of platelet 5-HI to a similar extent as ridogrel alone; plasma prostanoids were affected as with aspirin alone. 6. Combined administration of ketanserin and ridogrel virtually eliminated the collagen-induced platelet adhesion/aggregation response and release of 5-HI; prostanoids were affected as with ridogrel alone. 6. Combined administration of ketanserin and ridogrel virtually eliminated the collagen-induced platelet adhesion/aggregation response and release of 5-HI; prostanoids were affected as with ridogrel alone. 7. The results indicate that the interplay between 5-HT and arachidonic acid metabolites is causally involved in the platelet reaction to activation induced by collagen in cats in vivo.

Continuous conjunctival oxygen tension (PcjO2) monitoring for assessment of cerebral oxygenation and metabolism during carotid artery surgery.

The clinical value of noninvasive continuous monitoring of conjunctival oxygen tension for assessment of cerebral perfusion during carotid endarterectomy performed under general anaesthesia has been evaluated. The patients (n = 17; mean age 62.5 +/- 1.7 years) were monitored as follows: conjunctival oxygen tension (PcjO2); internal jugular venous oxygen tension at the skull base level (PcijvO2); arterial blood pressure; arterial and internal jugular venous blood gases; acid-base data and lactate, pyruvate levels; end-tidal CO2 concentration. The mean preanaesthetic PcjO2 level of 4.86 +/- 0.40 kPa was significantly lower than PaO2(PcjO2)/PaO2 ratio of 0.48). Following anaesthesia, a larger PcjO2-PaO2 gradient (ratio 0.32) was seen in spite of the hyperoxic situation (FiO2 = 0.40) due to vasoconstriction induced by slight hypocapnia (reduction of PaCO2 from 5.13 +/- 0.08 to 4.64 +/- 0.10 kPa). The carotid artery crossclamping resulted in a rapid and pronounced decrease of PcjO2, while PcijvO2 remained unchanged. No relationship between PcjO2 and stump pressure was found, while a significant correlation (P less than 0.02) between PcjO2 and lactate in effluent venous blood from the brain was demonstrable. It is concluded that PcjO2 monitoring seems a clinically useful trend indicator of cerebral perfusion in the individual patient. Due to large interindividual variations in basal PcjO2 readings and in PcjO2 changes during carotid artery clamping, however, transconjunctival oxygen tension monitoring does not seem to allow early and accurate recognition of impending cerebral ischaemia during carotid endarterectomy, and its routine use therefore seems of limited value.

Effects of the vagus nerves on gastric motility and release of vasoactive intestinal polypeptide in the anaesthetized lamb.

1. Effects of the vagus nerves on the activity of the reticulo-omasal orifice (ROO) and the abomasum and the concentration of vasoactive intestinal polypeptide (VIP) in gastric and intestinal venous effluent were studied in anaesthetized lambs. 2. Both excitatory and inhibitory effects of the vagus on the ROO and abomasum were demonstrated. Excitation of activity of the ROO was dominant with stimulation at lower frequencies (less than or equal to 20 Hz) whereas higher frequencies (50-70 Hz) caused inhibition. 3. Inhibition but not stimulation of the ROO and abomasum were obtained with vagal stimulation after administration of atropine (200-500 micrograms kg-1 I.V.). 4. The concentration of VIP in gastric venous effluent blood rose by 41 +/- 10 pmol l-1 (n = 13) within 90 s of the start of stimulation (10 Hz, 5 ms pulse duration, 10 V for 180 s) of a vagus nerve and fell within 60 s of its cessation to or near basal levels. The effects of vagal stimulation in changing the concentration of VIP were not altered by administration of atropine. 5. The concentration of VIP in the intestinal venous effluent increased during stimulation (10 Hz, 5 ms pulse duration, 10 V, for 180 s) of a vagus nerve by 15 +/- 5 pmol l-1 (n = 11) and remained elevated for more than 10 min after the stimulus had ceased. 6. The changes in gastric motility and in concentration of VIP in gastric and intestinal venous blood with stimulation of the vagus nerves have been discussed in relation to similar alterations which occur in lambs during sucking of milk.

Evidence from O2 uptake measurements for Na+ -K+ -2 Cl- co-transport in the rabbit submandibular gland.

There is evidence that the production of primary saliva by acinar cells is a consequence of Na+ -Cl- co-transport but more recently it has been proposed that in fact Na+ -K+ -2 Cl- co-transport is responsible. The latter would be energetically more efficient and the present experiments were designed to measure the stoichiometry of acinar secretion in order to distinguish between these two mechanisms. Submandibular salivary glands from anaesthetised rabbits were isolated vascularly and oxygen consumption measured from the oxygen content of arterial inflow and venous effluent blood and the total flow through the gland. Measurements were made in the steady-state at rest and during different secretion rates induced by parasympathetic nerve stimulation. The rate of sodium transport across the acinar and ductal epithelium was determined from plasma and salivary sodium concentration and salivary flow rate. Multiple regression analysis of this data showed that 22.1 mol Na+ was secreted per mol O2 consumed while 11.9 mol Na+ was reabsorbed per mol O2 consumed. Since acinar secretion is energetically about twice as efficient as ductal absorption, a mechanism for Na+ transport other than that for tight epithelia must be involved. Na+ -K+ -2 Cl- co-transport is thus more likely than Na+ -Cl- and it is suggested that Na+ -K+ -2 Cl- co-transport is the main mechanism involved in salivary acinar secretion.

Inert gas wash-out from tissue: Model analysis

Model simulations on variously arranged two-compartment models are performed to provide a basis for interpreting the observed non-monoexponential (or non-linear logarithmic) wash-out time courses of inert gases from tissue. The variables considered are: blood flow, tissue volume, solubility of gas in tissue and blood, and diffusive conductances (diffusing capacities) for tissue/blood gas transfer and for gas transfer between tissue compartments. The wash-out is studied in terms of both mean tissue partial pressure and effluent venous blood partial pressure. Diffusion limitation within a tissue-blood capillary unit is shown to produce logarithmic wash-out rates which increase or decrease during wash-out, depending on the functional structure of the unit. On the other hand, in a system consisting of dissimilar tissue-blood capillary units arranged in parallel, the logarithmic wash-out rate decreases with wash-out time. It is shown that the conventional analysis of nonlinear logarithmic wash-out may overestimate tissue perfusion or the extent of its inhomogeneity.

Changes in Vascular Resistance in the Feline Urinary Bladder in Response to Bladder Filling

The effect of bladder filling on vesical blood flow was studied in anesthetized cats. The pelvic nerves were sectioned and the bladder venous outflow was recorded by a drop counter. Filling of the bladder induced a sustained increase in vesical blood flow, related to the bladder volume. Thus, injection of 10, 20 and 30ml. of saline decreased bladder vascular resistance by 27, 47 and 52 per cent, respectively. This vasodilatation occurred despite a steady state increase in bladder pressure by 25 to 50cm. H2O. Upon emptying of the bladder, blood flow decreased towards control values. The response was unaffected by α-adrenoceptor blockade (phentolamine), β-adrenoceptor blockade (propranolol) and muscarinic cholinoceptor blockade (atropine). The vasodilatation was eliminated by papaverine, indicating that the response was not due to a rearrangement of the vessels at large bladder volumes. The vasodilatation was not associated with any increase in the concentrations of vasoactive intestinal polypeptide (VIP) or substance P in the venous effluent blood. Indomethacin significantly reduced the vasodilatation induced by filling. We conclude that local mechanisms, possibly involving release of prostaglandins, are involved in the vasodilatation following bladder distension.