Occult glove perforation during ophthalmic surgery. We examined the latex surgical gloves used by 56 primary surgeons in 454 ophthalmic surgical procedures performed over a 7-month period. Of five techniques used to detect pinholes, air inflation with water submersion and compression was found to be the most sensitive, yielding a 6.80% prevalence in control glove pairs and a 21.8% prevalence in postoperative study glove pairs, for a 15.0% incidence of surgically induced perforations (P = 0.000459). The lowest postoperative perforation rate was 11.4% for cataract and intraocular lens surgery, and the highest was 41.7% for oculoplastic procedures. Factors that correlated significantly with the presence of glove perforations as determined by multiple logistic regression analysis were oculoplastic and pediatric ophthalmology and strabismus surgical procedures, surgeon's status as a fellow in training, operating time, and glove size. The thumb and index finger of the nondominant hand contained the largest numbers of pinholes. These data suggest strategies for reducing the risk of cross-infection during ophthalmic surgery.


Rehabilitation and insurance relationships. The ultimate goal of the health care process is to restore an individual to a maximal level of function within the constraints of the existing pathology. It is unfortunate, however, that this potential is often underestimated so that the individual is deprived of reintegration in society. Rehabilitation is the unfinished business of medicine. Reasonable goals need to be identified and clear cost benefit projections need to be established regarding any intervention. To decrease an individual's dependency means that the lifetime cost of care of the individual decreases dramatically. Physical independence and cognitive competency reestablish for the patient optimal levels of health care that need to be facilitated, and involve integrated, innovative insurance liaisons to assure achieving this goal.


Reference interval and critical difference for canine serum fructosamine concentration. The purposes of the study were to obtain a reference interval and to calculate the critical difference between two analytical results for canine serum fructosamine concentration. To obtain a reference interval, the serum fructosamine concentration was measured in blood samples from 29 adult dogs after a 15-h fasting period. To calculate the critical difference, blood samples from 20 apparently clinically healthy dogs were collected once weekly for five consecutive weeks, and the total variance of the analytical results was divided into the component of variance between dogs (S2inter), the component of variance for weeks within dogs (S2intra) and the component of variance for measurements (S2anal), using nested analysis of variance. The critical difference was then calculated from S2intra and S2anal. The main conclusions are in summary: The reference interval for canine serum fructosamine concentration is 258.6-343.8 mumol/L, and the critical difference between two consecutive measurements on a week-to-week basis is 32.4 mumol/L. The critical difference may be used as a guideline to indicate potentially important changes in the serum fructosamine concentration, though the analytical results should not be assessed by the critical differences alone, but should also be compared to the corresponding reference intervals.


Gastrointestinal helminths and lungworms of French dairy goats: prevalence and geographical distribution in Poitou-Charentes. A study was conducted on 81 dairy-goat farms in western France. Faecal samples were collected once and 31 culled goats were necropsied in order to assess their gastrointestinal and respiratory helminth species, their prevalences and the intensities of infection as well as their geographical distribution in six subregions of the surveyed area. Fifteen species of helminth were recovered. The most frequent were Muellerius capillaris, Trichostrongylus colubriformis and Teladorsagia circumcincta with a prevalence more than 90%. The two latter species represented respectively 50.9% and 30.2% of the total worm burden of the digestive tract. Other prevalent species were Haemonchus contortus, Strongyloides papillosus, Trichuris spp., Moniezia spp. and Dicrocoelium lanceolatum. The occurrence of Dicrocoelium lanceolatum was related to the presence of calcareous or alkaline soils and restricted to the south of the surveyed area. No area at particular risk was defined for the other helminth infections, probably because of the relative homogeneity in climatic and topographic conditions of the six agricultural regions.


Parenchymal injury and biliary obstruction in relation to photosensitization in sporidesmin-intoxicated lambs. The hepatic changes were compared in lambs photosensitized or not photosensitized after exposure to sporidesmin. Injury to both the parenchyma and the biliary system was more severe in the photosensitized than in the non-photosensitized lambs. The activities of gamma-glutamyltransferase and glutamate dehydrogenase, and the total, conjugated and unconjugated bilirubin concentrations were significantly higher in sera from the photosensitized than from the non-photosensitized lambs. Hepatic glycogen levels were decreased in both the photosensitized and the non-photosensitized lambs, but were significantly lower in the former. Hence it is possible that lesions in hepatocytes contribute to retention of phylloerythrin and so to photosensitization.


The vascular responses of the isolated perfused bovine external ear to exogenous histamine and 5-HT (serotonin). The responses of the vasculature of isolated, non-sensitized, bovine external ears to histamine and serotonin (5-HT) were evaluated while they were being perfused with Krebs-Henseleit solution. Histamine (10(-5) mol/L to 5 x 10(-3) mol/L) and 5-hydroxytryptamine (5-HT) (10(-9) mol/L to 10(-2) mol/L) caused increased vascular resistance. Mepyramine (10(-7) mol/L), cimetidine (10(-5) mol/L) and atropine (10(-6) mol/L) inhibited the responses to histamine. The responses to 5-HT were inhibited by methysergide (10(-9) mol/L) and potentiated by morphine (10(-5) mol/L). These results suggest the presence of H1 and H2 histamine, and 5-HT receptors in bovine auricular vessels, all of which cause net vasoconstriction.


The influence of age on the pharmacokinetics of aditoprim in pigs after intravenous and oral administration. Some pharmacokinetic parameters of aditoprim were determined in 3- and 6-month-old pigs. After intravenous administration of 5 mg/kg body weight, the mean total body clearance of the older pigs was smaller than that of the younger pigs. This difference was not reflected in the elimination half-life. After oral administration of 5 mg/kg body weight, the mean absorption rate constant was smaller and the mean absorption half-life was longer in the older pigs. The age-related changes in the pharmacokinetics of aditoprim were not sufficiently pronounced to suggest the necessity of modifying the oral dosage regimen in pigs of this age range. The favourable pharmacokinetics of aditoprim in pigs (large apparent volume of distribution, long elimination half-life and high bioavailability) may permit introduction of this drug into swine practice, after safety and residue depletion studies.


The effect of protein binding on ivermectin uptake by bovine brain microvessel endothelial cells. The effect of albumin binding on ivermectin uptake and transfer across the endothelial component of the blood-brain barrier (BBB) was determined with an in vitro model comprised of bovine brain microvessel endothelial cell (BMEC) monolayers. Cellular uptake of ivermectin was limited in the absence of albumin and 90% inhibited in the presence of 10% albumin. Cell membrane association of ivermectin, as followed by fluorescent probe labelling, was observed only at high (micromolar) concentrations of the drug. Membrane association was about 75% inhibited in the presence of albumin. Similarly, transfer across BMEC monolayers was restricted, equivalent to that of BBB impermeant markers. Unlike the uptake studies, however, albumin had little effect on the transfer of ivermectin across BMEC monolayers. These results support recent in vivo findings on the distribution of ivermectin into the brain and suggest that ivermectin has only a limited affinity for the endothelial component of the normal BBB.


Food intake and rumen motility in dwarf goats. Effects of some serotonin receptor agonists and antagonists. The serotonergic regulation of feeding behaviour has not so far been studied in ruminants. Therefore, the effects of some serotonin (5-HT) receptor agonists and antagonists on food intake and forestomach motility were studied in dwarf goats. Goats ate less food when treated intravenously (IV) with the 5-HT precursor 5-HTP (25 micrograms, 50 micrograms or 100 micrograms kg-1 min-1 over 15 min) than when they were treated with 5-HT (which does not pass the blood-brain barrier) or with saline. Accordingly, IV dexfenfluramine infusions (50 micrograms or 100 micrograms kg-1 min-1 over 15 min), which induces release of brain 5-HT, also led to dose-related reductions in food intake. In contrast, no anorectic effects were observed after IV infusions with the selective 5-HT reuptake inhibitor fluoxetine (100 micrograms kg-1 min-1 over 15 min), the selective 5-HT1A agonist 8-OH-DPAT (0.5 micrograms kg-1 min-1 over 15 min), or eltoprazine (4 or 8 micrograms kg-1 min-1 over 15 min), a mixed 5-HT1A/5HT1B receptor agonist. None of the 5-HT antagonists tested gave any increase in food consumption in this model. Interestingly, the non-selective 5-HT receptor antagonist methysergide (360 micrograms/kg IV) reduced food intake. This effect was most noticeable at 3 h after injection. The 5-HT3 receptor antagonist ondansetron (IV 10 micrograms kg-1 min-1 over 15 min) and the peripheral 5-HT2 receptor antagonist xylamidine (IV 100 micrograms kg-1 min-1 over 10 min) failed to modify food intake. These results provide evidence for central serotonergic involvement in the control of feeding. However, this control system differs markedly in goats and rodents. Dexfenfluramine, 5-HTP and eltoprazine administered at similar dose rates to those used in the food intake experiments induced some clinical signs including inhibition of forestomach contractions. These results, together with our earlier in vivo and in vitro observations, suggest that the inhibitory effects of serotonin receptor agonists on forestomach contractions are due to interactions with both peripheral and central serotonergic receptors. The change in smooth muscle tension, which leads to a change in the signals transmitted via vagal afferents to the central nervous system, appears not to modify feeding behaviour in dwarf goats.


Technical and methodological requirements for reliable haemodynamic measurements in the unsedated calf. The purpose of this study was (1) to evaluate the technical and methodological problems associated with invasive haemodynamic measurements in unsedated cattle; (2) to assess the reproducibility of such measurements both within and between days; and (3) to compare the values with those previously reported. Twenty-one healthy calves, aged from 5.5 to 12 months, were studied. The central venous, the right ventricular, the pulmonary arterial, the pulmonary capillary wedge and the systemic arterial pressures were obtained by means of fluid-filled catheters, and the cardiac output was measured by the thermodilution technique. The heart rate, the stroke volume, the pulmonary and systemic vascular resistances and the pulmonary and systemic ventricular workloads were calculated. An adverse reaction, consisting of severe pulmonary hypertension, tachycardia, tachypnoea and transient weakness, occurred in 7 calves during the catheterization procedures. Such a reaction might be due to a local reflex induced by stimulation of mechano-receptors by the catheter tip. It should be avoided by reducing the manipulation of the catheter as much as possible and by inflating the tip of the balloon when moving it forwards. A comparison of the vascular pressures with those previously reported was difficult because of methodological or technical limitations, such as, for instance, a lack of standardization of the baseline. The reproducibility of the haemodynamic measurements obtained was satisfactory, in contrast to previous studies performed in conscious animals. This was attributed to our animals being better trained to the experimental conditions and emphasizes the importance of reducing mental stress in obtaining reliable haemodynamic measurements in unsedated and potentially uncooperative animals.


[The dilemma of syndrome X].

[The dilemma of syndrome X]. The term X syndrome involves several dilemmas. The terminological dilemma is that this term is used to describe microvascular angina pectoris, as well as Reaven's metabolic-hormonal syndrome and our 5H syndrome [association of hyperinsulinism with arterial hypertension, hyperlipoproteinaemia, hyperglycaemia (NIDDM) and hirsutism]. It is probable that the coronary X syndrome is frequently conditioned by the hormonal-metabolic X syndrome. The pathogenetic dilemma is that it is not clear why in microvascular angina pectoris the coronary circulation does not possess an adequate reserve for vasodilatation during exercise or in response to some pharmacological stimuli. This could condition hyperinsulinism in hypertonic subjects with NIDDM with an early disorder of paracrine endothelial relaxation mechanisms (EDRF-NO), with a predominance of vasoconstrictor mechanisms (endothelin-1). In Reaven's syndrome X it is not clear whether insulin resistance of the postreceptor type is a primary inborn phenomenon which is compensated by insulin hypersecretion or whether it is a secondary phenomenon, which develops ex post and by which the target tissues defend themselves against an excessive effect of insulin (e.g. down regulation receptors) in primary disorders of its secretion.


[Long-term effect of drug therapy in supraventricular dysrhythmia evaluated by diagnostic esophageal cardiac pacing]. The authors investigated the long-term therapeutic effect in 33 patients after a medicamentous reversal of recent atrial fibrillation and 18 patients after reversal of an attack of supraventricular tachycardia (SVT). None of the patients had signs of cardiac failure. A change to sinus rhythm was achieved in the group with atrial fibrillation by digoxin with verapamil, in the group with SVT by verapamil. Treatment was then adjusted, based on testing by means of diagnostic oesophageal pacing. The basic drug for long-term use in atrial fibrillation was digoxin either as monotherapy or in combination; the majority of patients with SVT had verapamil treatment. At the end of the period 24.5 +/- 8.4 (12-36) months after the reversal in the group with atrial fibrillation 8 (25.8%) had permanent atrial fibrillation or different therapy on account of atrial fibrillation; short-term attacks of palpitation were recorded during the investigation period in 16 (51.6%), and the remaining 7 patients (22.6%) had no complaints. In the group with SVT none of the patients was hospitalized on account of persisting dysrhythmia and it was not necessary to alter treatment. Testing treatment of supraventricular dysrhythmias by means of diagnostic oesophageal pacing is of prognostic importance and the authors consider it the method of choice.


[Directed sympathetic denervation of the heart--an actual therapeutic method or myth?]. Aimed sympathetic denervation of the heart, i. e. bilateral removal of the stellate ganglion Th 1-Th 3 was used in Czechoslovakia since 1981 in eight patients who were followed up for prolonged periods. The patients were operated on account of angina at rest which caused deterioration of the quality of their lives. In case of extensive affection of the coronary arteries surgical treatment was not feasible and pharmacological treatment did not bring relief. At the time of a five-year check up 7 patients survived. All patients reported relief and their efficiency during ergometry improved. Four of the patients worked for several years. The authors discuss whether the above treatment is still justified.


[Hyperinsulinism and the coronary syndrome]. In one third of patients who suffered an infarction NIDDM and arterial hypertension are present. In the absolute majority of patients with IHD, as apparent from the IRI and C-peptide response after a glucose load, hyperinsulinism is present. The blood sugar response can have the character of diabetes or of impaired glucose tolerance, the curve may be very flat or normal while the IRI and C-peptide response are excessive. Hyperinsulinism has a hypersecretory origin as suggested by the concurrently elevated C-peptide level but also reduced insulin utilization in the liver and peripheral target organs. Hyperinsulinism is thus a regular associated phenomenon of IHD and is a special risk factor independent on hyperglycaemia and associates with the other main risk factors of IHD such as arterial hypertension, HPLP (android obesity), hyperglycaemia (NIDDM) and hirsutism as a manifestation of a hyperandrogenic state in the female organism with the syndrome of polycystic ovaries. Hyperinsulinism plays an indirect role in the pathogenesis of coronary syndrome via the main risk factors (5H syndrome--hyperinsulinism, hypertension, HPLP, hyperglycaemia, hirsutism) and also directly by its action on endothelial paracrine mechanism of the coronary circulation where in the early stage vasoconstrictor factors predominate (endothelin-1, PGF2-alpha) over physiological vasodilatating factors (EDRF-NO, PGE2, PGI2) and this leads then to functional spasms. It seems that also the coronary X syndrome develops very frequently on the background of the hormonal metabolic X syndrome or the 5H syndrome.


[Gestational diabetes mellitus and disorders of glucose tolerance in pregnant women with essential hypertension]. The authors revealed during dispensarization of pregnant women suffering from essential hypertension that the disease is relatively frequently associated with some metabolic disorders, i. e. obesity, gestational diabetes or impaired glucose tolerance. They draw attention to a similarity with Reaven's syndrome in non-pregnant women. The authors recommend to screen for diabetes all obese pregnant women and those with hypertension to detect an impaired glucose metabolism and prevent foetopathies in neonates of thus affected mothers. The authors consider obesity one of the subsidiary criteria in the differential diagnosis of essential hypertension and preeclampsia.


[24-hour effect and tolerance for perindopril (Prestarium) in mild and moderate essential hypertension]. A multicentre investigation of antihypertensive treatment by perindopril was conducted in three centres of Czechoslovakia. The investigation comprised 51 patients with mild to moderate essential hypertension (diastolic pressure 95-115 mmHg) who were taking perindopril for a period of three months, the initial dose being 4 mg (1 tablet per day). If the blood pressure did not drop to normal levels (dBP < 90 mmHg), the dose was increased to 8 mg after 1 month and if after 2 months of treatment the blood pressure did not reach normal levels, a thiazide diuretic was added. The antihypertensive effectiveness was assessed 24 hours after ingestion of the drug. The blood pressure declined significantly already after the first month of monotherapy with perindopril. It declined then significantly up to the end of the 3rd month of treatment. The systolic blood pressure declined by 9.4 mmHg after the first month of monotherapy with perindopril and by 15.9 mmHg after 3 months of treatment. The diastolic blood pressure declined by 8.1 mmHg after one month of monotherapy and by 12.4 mmHg at the end of three-month treatment. The clinical and biological tolerance was excellent and only one patient discontinued treatment for reasons not related to the administered drug. Twenty-four hour monitoring of the blood pressure also proved the favourable effect of perindopril.


[The effect of self-monitoring on perinatal outcome in insulin therapy of diabetic women during pregnancy]. The authors compare perinatal results in insulin treated diabetic women who were followed up in the out-patient department of the Second Gynaecological and Obstetric Clinic of the Masaryk University in Brno. In 1980-1984 148 pregnant diabetic women were followed up and in 1985-1991 when self-monitoring has become part of comprehensive treatment of pregnant diabetics in our department, 279 women were followed up. The patients were monitored during pregnancy by a diabetologist and gynaecologist. Other members of the team are a paediatrician, geneticist, psychologist, dietitian, and educational nurse. As to laboratory parameters, short-term and long-term compensation of diabetes were monitored. Systematic preconception preparation of diabetic patients, with an attempt to achieve normal blood sugar levels and genetic examination of both partners from part of comprehensive care. The results achieved in our department revealed a statistically significant decline of the incidence of foetopathies in neonates at the 5% level of significance and a decline of hypoglycaemia in foetuses at the 0.1% level.


[Prenatal preparation--a part of the care of young female diabetic patients]. In the Second Gynaecological and Obstetric Clinic in Brno there has been for more than 15 years a specialized department for treatment of diabetes during pregnancy. The department monitors and treats pregnant diabetics from approximately two thirds of the former South Moravian region, i.e. patients with diabetes assessed before pregnancy as well as those where an impaired carbohydrate metabolism was detected during pregnancy. The authors submit an account of the organization of team work in their out-patient department.


[Ablation of the accessory bundle of Kent using radiofrequency current in a 17-year-old boy with type A Wolff-Parkinson-White syndrome]. The authors describe the in Czechoslovakia so far not used therapeutic catheterization ablation method of the accessory atrioventricular pathway in patients with tachycardia with the Wolff-Parkinson-White syndrome. They performed successful ablation of the accessory bundle of Kent located by electrophysiological examination behind the left atrium and left ventricle in a 17-year-old boy with WPW Syndrome type A. They severed the accessory pathway by radiofrequency current using a catheter inserted from a retrograde approach into the left ventricle beneath the mitral valve. After ablation the boy suffers no longer from tachycardia, the ECG tracing in devoid of preventricular preexcitation, although he has no medicamentous treatment.


[Subphrenic abscess--a complication of intensive cytostatic treatment in acute leukemia]. The authors describe the slow development of an abscess in the left subphrenic space in a patient with acute myeloid leukaemia. The patient suffered several months before the diagnosis was established from pain in the left subcostal region and was on account of this pain examined repeatedly by clinical methods and sonography. During the last sonographic examination in this area a hypoechogenic formation was detected. The diagnosis was than established more accurately by computer tomography by visualization of the abscess cavity. The case-history and relatively thick wall of the cavity suggested a long-term process. The abscess cavity was evacuated surgically, however, the patient suffered a relapse later and died from septicaemia. In the discussion the authors analyze the problem of development of metastatic abscesses in leukopenic patients, early diagnosis and treatment.


[A new concept of atherogenesis: the role of oxygen radicals]. The author discusses a new concept of atherogenesis which is based on older hypotheses (lipid infiltration hypothesis), theory of impaired endothelial barrier, role of immunological and thrombocytic factors) and supplements it by new findings on the role of oxygen radicals in the development of atherosclerosis. The atherogenity of native low density lipoproteins (LDL) is enhanced substantially by their post-secretory changes caused by the action of free oxygen radicals on unsaturated fatty acids in the outer layer of LDL particles. The development of lipid peroxides on the surface of LDL leads to a changed orientation of the metabolism of LDL particles: oxidized LDL are not bound to classical LDL receptors in the liver but to so-called scavenger receptors in monocytes and macrophages. Lipid accumulation in monocytes, not controlled by feedback mechanisms, leads to formation of foam cells which are the basis of atheromatous plaques. This process can be influenced by antioxidants which block the formation of lipid peroxides. The new concept is particularly important for the population of Czechoslovakia and other formerly totalitarian countries where deterioration of the environment occurred and concurrently also adverse dietary changes (high fat, sucrose and spirit intake, low vegetable and fruit consumption). It is probable that an increased intake of antioxidants (ascorbic acid, carotenes, tocopherols, bioflavonoids, selenium etc.) would, along with other corrective provisions, suppress the steep increase of cardiovascular diseases.


[Insulin resistance as a risk factor in atherosclerosis]. Clinical complications of atherosclerosis are the main cause of mortality in the most advanced countries. Epidemiological data supported by extensive perspective investigations suggest the role of hyperinsulinaemia as an independent risk factor in the development of atherosclerosis. The authors summarize the trends in research of insulin resistance, its possible diagnostic and treatment.


[Therapy of developed atherosclerosis].

[Therapy of developed atherosclerosis]. Atherosclerosis is a reversible process and is an excessive response to damage of the vascular wall. It comprises a number of cellular processes and humoral modulations/regulations the pathogenesis of which is intensively studied. The reversibility of atherosclerosis changes fundamentally the approach to its prevention and therapy, and the effectiveness of anti-aggregation and antihyperlipaemic therapy has been proved. Other preventive and therapeutic procedures are intensively studied.


[New pathogenic findings on the mechanism of heart failure and their importance in therapy]. Activation of the neuroendocrine system participates in a decisive way in the pathogenesis of cardiac insufficiency. The authors draw attention to the basic therapeutic impact of the phenomenon of so-called "down-regulation" of myocardial beta-adrenoreceptors which takes place during chronic cardiac insufficiency, contrary to acute cardiac failure. The authors deal from the pathogenetic aspect with different groups of positive inotropic substances used in the treatment of cardiac insufficiency with special attention to digoxin and cardiac glycosides. They evaluate the effect of digoxin as a neurohumoral modulator in acute and chronic cardiac insufficiency. Its potential asset is the positive modulation of excessive neuroendocrine activation and the favourable effect on cardiopulmonary baroreflexes. The authors explain also why at present among positive inotropic substances in the treatment of cardiac insufficiency only cardiac glycosides are indicated. They mention the theoretical possibility to extend the indication of digoxin to retard the progressive deterioration of cardiac function in asymptomatic dysfunction of the left ventricle caused by the neurohumoral modulating effect of digoxin.


[Pathogenesis and fundamentals of treatment in acute myocardial infarct]. Recent findings which extend and render more accurate factors which influence the genesis and course of experimental and clinical acute myocardial infarction change in a significant way the treatment and management of patients with acute infarction. The authors discuss the basic factors and complex of interactions which play the most important part in the pathophysiology of acute infarction. They pay attention also to other factors which can influence in a significant way the course of acute infarction and the patient's prognosis (residual thrombosis, rethrombosis, extension, expansion of the infarction and remodelling of the left ventricle). In recent years it has become obvious that the most important determinants of the final extent of acute infarction is the duration and site of occlusion of the coronary artery, its recurrence and degree (magnitude), the condition of the collateral circulation and the consumption and requirements of the myocardium with respect to oxygen during the time of occlusion. The contemporary reperfusion-thrombotic era of treatment of acute myocardial infarction and management of the patient is based on logical multifactorial and to a certain extent preventive, active or aggressive approach and omission of all even potentially harmful approaches. All therapeutic operations (incl. invasive and radical ones) must be consistent with contemporary pathogenetic findings. They should be implemented before possible reinfarction, sudden cardiac death or treatment which has only little effect on heart failure which may be associated also with undesirable expansion of the infarction and remodelling of the left ventricle after acute myocardial infarction. Recent pathophysiological findings and the development of treatment of myocardial infarction are a challenge for cardiologists of the nineties to make use of these important findings in the treatment of patients with myocardial infarction and to ensure for them optimal treatment and management.


Ester hydrolysis and conjugation reactions in intact skin and skin homogenate, and by liver esterase of rabbits. 1. Procaine, 2-chloroprocaine, ethyl aminobenzoate and methyl salicylate were added at various concentrations to liver esterase, supernatant of skin homogenate, or single-pass perfused ears of rabbits. 2. Vmax of product formation by purified liver esterase correlated with the rank order of the distribution coefficients (n-octanol/buffer) of the substrates and ranged between 11 and 1100 pmol/min per micrograms protein. Km values were between 20 and 50 microM. 3. No correlation was observed when the apparent enzyme kinetics, calculated by non-linear adaptation, were compared with each other after substrate administration to skin, arterial influx, or incubation with skin homogenate. 4. An acid labile conjugate of ethyl 4-aminobenzoate was found, mainly during arterial perfusion and in supernatant of skin homogenate, after administration of ethyl 4-aminobenzoate. 5. Acetamidobenzoic acid was observed in quantities of about 10% of the free 4-aminobenzoic acid during dermal or arterial application of procaine. This metabolite was not found with ethyl 4-aminobenzoate. 6. The results from isolated rabbit ear perfusion differ quantitatively and qualitatively with those obtained from supernatant of skin homogenate or purified liver esterase.


Metabolism of methapyrilene by Fischer-344 rat and B6C3F1 mouse hepatocytes. 1. Suspension cultures of freshly isolated F344 rat and B6C3F1 mouse hepatocytes were compared for their ability to transform various concentrations of methapyrilene (MP). 2. MP metabolites were isolated and purified by h.p.l.c., and were identified by comparing their chromatographic and mass spectral properties with those of authentic standards. 3. Both rat and mouse hepatocytes transformed MP to tentatively identified 2-thiophenecarboxylic acid (I), and definitively identified mono-N-desmethyl methapyrilene glucuronide (II), methapyrilene glucuronide (III), methapyrilene N-oxide (V), and mono-N-desmethyl methapyrilene (VII).


Biotransformation of tri-substituted methoxyamphetamines by Cunninghamella echinulata. 1. Four trimethoxyamphetamine analogues were incubated with the filamentous fungus Cunninghamella echinulata. 2. 2,4,5-Trimethoxyamphetamine and 2,5-dimethoxy-4-ethoxyamphetamine were poorly metabolized by C. echinulata ATCC 9244 and C. echinulata var. elegans ATCC 9245. 2,5-Dimethoxy-4-(n)-propoxyamphetamine was mainly metabolized through N-acetylation and O-dealkylation with minor amounts of several aliphatic hydroxylation metabolites formed. 2,5-Dimethoxy-4-methylthioamphetamine was extensively metabolized to the corresponding sulphoxide. 3. 2,5-Dimethoxy-4-methylthioamphetamine metabolism was inhibited by ethanol and quinidine. Sparteine did not inhibit the formation of the sulphoxide and may have shunted the substrate through alternate metabolic pathways. 4. Incubation conditions can affect the rate and extent of fungal biotransformation of 2,5-dimethoxy-4-methylthioamphetamine, and influence dextrose utilization, ammonia formation and pH.


N-benzylimidazole-mediated changes in hepatic drug-metabolizing enzyme activities in Ah-responsive and Ah-non-responsive mice. 1. The inductive effect of N-benzylimidazole (NBI) on hepatic microsomal and cytosolic drug-metabolizing enzyme activities in aryl hydrocarbon (Ah)-responsive C57BL/6N (B6) and Ah-non-responsive DBA/2N (D2) mouse strains was determined and compared with that caused by beta-naphthoflavone (BNF). 2. Relative Ah-responsiveness of the two strains was confirmed by measurement of BNF-induced ethoxyresorufin deethylase (EROD) activity and ELISA immunoquantification. BNF markedly induced EROD activity only in the Ah-responsive B6 mouse strain (65-fold increase). 3. NBI (150 mg/kg per day for 3 days) increased cytochrome P450 concentration similarly in both strains (40 and 60% in B6 and D2 strains, respectively). Compared with BNF treatment of the B6 strain, increases in EROD activity following NBI treatment were only minor. In addition, EROD activity increases were greater in the Ah-nonresponsive D2 strain (300%) than in the Ah-responsive B6 strain (100%) suggesting the possibility of an induction mechanism different from that of recognized Ah receptor agonists. 4. Induction of UDP-glucuronosyltransferase activity (p-nitrophenol acceptor) by BNF was greater in the Ah-responsive B6 strain than in the Ah-non-responsive D2 strain. NBI failed to induce this activity in either strain. 5. Induction of glutathione S-transferase activity towards 1-chloro-2,4-dinitrobenzene following NBI treatment occurred to the same extent (25% increase) as that seen following BNF treatment, in the Ah-responsive B6 strain. Neither xenobiotic affected this activity in the Ah-non-responsive D2 strain. 6. Although NBI is a major inducer, possessing Ah-like inducing properties in rat, it caused only minor changes in murine drug metabolizing enzymes.(ABSTRACT TRUNCATED AT 250 WORDS)


Rat liver microsomal metabolism of 2-halogenated 4-methylanilines. 1. Rat liver microsomal metabolism of 2-fluoro-, 2-chloro- and 2-bromo-4-methylaniline was investigated using h.p.l.c. Metabolites identified include products from side-chain C-hydroxylation (benzyl alcohols and benzaldehydes) and N-hydroxylation (hydroxylamines and nitroso derivatives). Aromatic ring hydroxylation was not a major reaction pathway. 2. A new type of microsomal metabolite was detected which was identified as a secondary amine, i.e. a halogenated N-(4'-aminobenzyl)-4-methylaniline. 3. In addition to these products azoxy, azo and hydrazo derivatives were formed. 4. Benzyl alcohols and halogenated N-(4'-aminobenzyl)-4-methylanilines were the major microsomal metabolites for all three 2-halogenated 4-methylanilines. 5. Quantification of the metabolite patterns demonstrated an influence of the type of halogen substituent on the rate of microsomal metabolism. The rate of side-chain C-hydroxylation increases in the order 2-fluoro-4-methylaniline < 2-chloro-4-methylaniline < 2-bromo-4-methylaniline. 6. The rate of N-hydroxylation increases from 2-bromo-4-methylaniline < 2-fluoro-4-methylaniline < 2-chloro-4-methylaniline. That 2-chloro-4-methylaniline is N-hydroxylated to a larger extent is in accordance with its greater mutagenicity, twice that of 2-bromo-4-methylaniline.


Effect of peroxisome proliferators on glutathione-dependent sulphobromophthalein excretion. 1. The effect of pretreatment of rats with the peroxisome proliferator 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) on sulphobromophthalein excretion from the isolated perfused rat liver has been investigated and compared with the effect of clofibrate which is also a peroxisome proliferator. 2. Rats fed 2,4,5-T at a dose of 0.25% in the diet showed a decrease in food intake, compared with controls and clofibrate-fed rats. 3. Treatment with either 2,4,5-T or clofibrate was associated with significant inhibition of the biliary excretion of unchanged, conjugated, and total sulphobromophthalein from perfused rat liver, compared with diet-matched controls. 4. There was a decrease in bile flow in the clofibrate-treated group, but not in the 2,4,5-T-treated group. 5. The results of the present study confirm previous studies that have shown an association between peroxisome proliferation treatment and inhibition of glutathione S-transferase-mediated sulphobromophthalein excretion.


Modulation of sulphobromophthalein excretion by ethacrynic acid. 1. Ethacrynic acid (EA), a phenoxyacetic acid diuretic, has similar effects to tienilic acid (TA) on rat liver glutathione S-transferase (GST) activity in vitro, using either 1-chloro-2,4-dinitrobenzene or sulphobromophthalein (BSP) as a substrate. EA inhibits the basic rat liver GST, with inhibition being greater with GST containing subunits 3 and 4 than with those containing subunits 1 and 2. 2. In vitro inhibitors of GST can inhibit biliary excretion of BSP in a perfused liver. 3. A single bolus dose of EA had no effect on BSP excretion from the isolated perfused rat liver, and this is most likely due to the rapid disappearance of EA from the perfusion media. Experiments using perfused rat liver indicated that a sustained high concentration of EA in the perfusion media has an inhibitory effect on the excretion of both unchanged and conjugated BSP. 4. A decrease in BSP excretion may not be an indicator of liver damage, but a consequence of GST inhibition.


Induction of rat hepatic mixed-function oxidases by acetone and other physiological ketones: their role in diabetes-induced changes in cytochrome P450 proteins. 1. To evaluate the role of ketone bodies in diabetes-induced changes in hepatic cytochrome P450 composition, rats were treated with acetone, 3-hydroxybutyrate or 1,3-butanediol. 2. Treatment with acetone enhanced the rat hepatic O-dealkylations of ethoxyresorufin and methoxyresorufin, and the hydroxylation of p-nitrophenol, but had no effect on lauric acid hydroxylation and ethylmorphine N-demethylation. Neither 3-hydroxybutyrate nor 1,3-butanediol modulated the metabolism of the above substrates. 3. Immunoblot analysis of hepatic microsomal proteins revealed that treatment with acetone increased the apoprotein levels of P4501A2, P4502B1/2 and P4502E1. 4. It is concluded that acetone is responsible, at least partly, for the diabetes-induced increase in hepatic microsomal P4501A2, P4502B1/2 and P4502E1 proteins but does not mediate the increases in the P4503A1 and P4504A1 proteins. On the basis of work from our own and other laboratories a mechanism for the diabetes-induced changes in hepatic cytochrome P450 proteins is proposed.


Xenobiotic-metabolizing enzyme activities in hybrid cell lines established by fusion of primary rat liver parenchymal cells with hepatoma cells. 1. The activities of xenobiotic-metabolizing enzymes were determined in hybrid cell lines (hepatocytoma, HPCT) which have been established by fusion of liver parenchymal cells from adult rat (PC) with cells from a Reuber hepatoma cell line (FAO). 2. Cytochrome P450 was not measurable spectrophotometrically in FAO and HPCT. P450-dependent conversion of testosterone was below the detection limit in FAO and only marginally present in HPCT. 3. Microsomal and cytosolic epoxide hydrolase, glutathione S-transferase and phenol sulphotranserase were low or even below detection limit in FAO. These enzyme activities were significantly higher in HPCT and correspond to about 1-10% the activities measured in PC. 4. 1-Naphthol UPD-glucuronosyl transferase activity was about 20% in FAO and about 100% in HPCT compared to PC. 5. Metabolic conversion of benzo[a]pyrene was low in FAO, high in PC, and intermediate in HPCT. The presented data, however, do not allow the conclusion whether this intermediate rate is catalyzed by similar P450 isoenzymes as in PC. 6. Due to the easily measurable phase II-metabolizing enzyme activities HPCT may, however, be useful for in vitro enzyme induction or repression studies.


Identification of urinary metabolites of human subjects acutely poisoned by p-chloronitrobenzene. 1. Urinary metabolites from human subjects acutely poisoned with p-chloro-nitrobenzene (p-CNB) were identified by g.l.c.-mass spectrometry. 2. Eight substances, namely, a very large amount of N-acetyl-S-(4-nitrophenyl)-L-cysteine, relatively large quantities of p-chloroaniline, 2-chloro-5-nitrophenol and p-chloroformanilide produced by pyrolysis of a substance originating from p-CNB, small amounts of 2-amino-5-chlorophenol and 2,4-dichloroaniline, and traces of p-chloroacetanilide and 4-chloro-2-hydroxyacetanilide, were detected in urine samples. 3. All of the absorbed p-CNB was metabolized prior to excretion, as the parent compound was not found in urine. 4. N-Acetylated metabolites of p-chloroaniline and 2-amino-5-chlorophenol, resulting from p-CNB by metabolism, were found in only one of eight individuals indicating that this pathway is weak or may be absent in some humans. 5. A scheme for the pattern of metabolic pathways of p-CNB is proposed, and chlorination was considered to be a possible novel metabolic pathway.


Extracellular intermediates of glucose metabolism: fluxes of endogenous lactate and alanine through extracellular pools in embryonic sympathetic ganglia. The flux rates of lactate and alanine in and out of the cells of an intact tissue, which cannot be measured directly because some of the released materials are reabsorbed, were determined by computer analysis of uptakes and outputs by the whole tissue in the presence of various concentrations of these substances. The outputs of labeled lactate and alanine from [U-14C]glucose and the uptakes of [U-14C]lactate and [U-14C]alanine were measured on intact sympathetic ganglia excised from 15-day-old chicken embryos. The volume and time constant of the extracellular space were measured using labeled lactate, alanine, and sucrose. Models, which mathematically described the cellular uptakes and outputs as functions of the extracellular concentrations, were used to predict the exchanges that would be observed on the whole tissue, and their parameters were adjusted for best fit to the actual observations. The fitted models were then used to calculate the fluxes in and out of the cells and the concentrations in the extracellular space. The following results were obtained: (1) Cellular uptakes of lactate and alanine were both well described by familiar Michaelis-Menten kinetics. (2) The cellular output of [14C]-lactate from [14C]glucose declined with increase in the extracellular lactate concentration, whereas the cellular output of [14C]alanine from [14C]glucose rose with the extracellular alanine concentration. (3) Half-saturation values for cellular uptake, determined from the fitted equations, were 0.45 mM for lactate and 1.17 mM for alanine, both several-fold lower than less relevant estimates for the whole tissue made directly from the uptake observations. (4) As much as 45% of the carbon in the glucose consumed was released into the extracellular space as lactate and alanine, but much of this was reabsorbed. Implications for brain metabolism are discussed.


Patienthood.

Patienthood. The experience of being a patient is valuable in learning how to become a humane physician. The behavior of students and house officers is modeled on their attending and senior residents. Interactions with patients will be more likely to resemble the example of senior physicians than any theoretical or ideal example, no matter how well taught.


Cerebral cortex ammonia and glutamine metabolism during liver insufficiency-induced hyperammonemia in the rat. Hyperammonemia has been suggested to induce enhanced cerebral cortex ammonia uptake, subsequent glutamine synthesis and accumulation, and finally net glutamine release into the blood stream, but this has never been confirmed in liver insufficiency models. Therefore, cerebral cortex ammonia- and glutamine-related metabolism was studied during liver insufficiency-induced hyperammonemia by measuring plasma flow and venous-arterial concentration differences of ammonia and amino acids across the cerebral cortex (enabling estimation of net metabolite exchange), 1 day after portacaval shunting and 2, 4, and 6 h after hepatic artery ligation (or in controls). The intra-organ effects were investigated by measuring cerebral cortex tissue ammonia and amino acids 6 h after liver ischemia induction or in controls. Arterial ammonia and glutamine increased in portacaval-shunted rats versus controls, and further increased during liver ischemia. Cerebral cortex net ammonia uptake, observed in portacaval-shunted rats, increased progressively during liver ischemia, but net glutamine release was only observed after 6 h of liver ischemia. Cerebral cortex tissue glutamine, gamma-aminobutyric acid, most other amino acids, and ammonia levels were increased during liver ischemia. Glutamate was equally decreased in portacaval-shunted and liver-ischemia rats. The observed net cerebral cortex ammonia uptake, cerebral cortex tissue ammonia and glutamine accumulation, and finally glutamine release into the blood suggest that the rat cerebral cortex initially contributes to net ammonia removal from the blood during liver insufficiency-induced hyperammonemia by augmenting tissue glutamine and ammonia pools, and later by net glutamine release into the blood. The changes in cerebral cortex glutamate and gamma-aminobutyric acid could be related to altered ammonia metabolism.


The iliac passion.

The iliac passion. "The Iliac Passion" traces a return from the new but busy and rapidly growing discipline of "bioethics" to its source in "fundamental philosophical inquiry." The dilemma between bioethics and medicine is examined in two ways. First, the philosophical concept of the "big question" is presented. If we ask of life or of human experience "What does it all mean?", the "it" needs to be defined, and what I propose to do is to "take on" the "it." In Part Two, the task of combining the medical-technical objectifying mode of thinking about patients, necessary to treat them effectively, with the ability to understand and sympathize with their pain and distress, is illustrated by means of a personal story or parable.


Restoring the patient's voice: the case of Gilda Radner. In the past few years, the medical case report has been studied as a document that evidences the way the patient and, by extension, the experiential and subjective aspects of an illness tend to be marginalized in contemporary medical theory and practice. First-person narratives about illness, our popular "pathographies," may in part represent our attempt as a culture to respond to this problem of "the vanishing patient." A rich source of information about patient experience, pathographies can be useful to us in locating specific issues in the medical enterprise that need understanding and perhaps require correction. Gilda Radner's It's Always Something demonstrates how two important issues--both neglected in the conventional medical history--powerfully affect the medical enterprise: the hopes, expectations, and wishes of the experiencing patient, and the perceived attitudes and demeanor of the patient's physicians. The restoration of patient and physician to the "history" is important not only because it reminds us of the personal dimension of the medical enterprise, but also because it alerts us to problems of attitude and action that bear directly on diagnosis, course of treatment, and the therapeutic transaction.


Humanities and medicine (a slightly dissident view). Programs for humanities and medicine are growing in a number of medical schools in the U.S.A. Proponents of the programs, which are intended to bring together humanists, scientists, physicians, and others, believe that broadening the background of physicians will put a more human face on the practice of medicine, despite its increasingly technological nature. There is little to support this premise, and its successes and failures are not measurable. There are reasons to support the programs, however, but they have more to do with what physicians like and want to do than with what is therapeutic for them.


Is the vulnerability of neurons in the substantia nigra of patients with Parkinson's disease related to their neuromelanin content? The contribution of neuromelanin (NM) to the pathogenesis of Parkinson's disease (PD) has long been suspected. In particular, a correlation has been reported between the estimated cell loss in the mesencephalic dopaminergic cell groups and the percentage of NM-pigmented neurons in these cell groups. To test whether the amount of pigment per cell is a critical factor or whether the presence of NM within a neuron is sufficient to account for the degeneration of dopaminergic neurons, the NM content was measured in each neuron from representative sections throughout the ventral mesencephalon of four controls subjects and four patients with PD. Intraneuronal NM was quantified by a densitometric method, using known amounts of synthetic melanin as standards. In control brains, the distribution of melanized neurons in the nigral complex showed a high proportion of lightly melanized neurons in the ventral tegmental area and the pars alpha and gamma of the substantia nigra (SN), whereas heavily melanized neurons were mostly located in the pars beta and lateralis of the SN. An inverse relationship was observed between the percentage of surviving neurons in PD compared with controls and the amount of NM they contain, suggesting that the vulnerability of the dopaminergic neurons is related to their NM content. Factors other than NM may be involved in the differential vulnerability of catecholaminergic neurons in PD. In particular, the constant topography of the cell loss suggests that cell position within the nigral complex is a key factor.


Unsaturated fatty acids esterified in 2-acyl-l-lysophosphatidylcholine bound to albumin are more efficiently taken up by the young rat brain than the unesterified form. The aim of the present study was to investigate whether unsaturated 2-acyl-lysophosphatidylcholine bound to plasma albumin is a relevant delivery form of unsaturated fatty acids to the developing brain. Twenty-day-old rats were perfused for 30 s with labeled palmitic, oleic, linoleic, and arachidonic acids in either their unesterified form or esterified in 2-acyl-lysophosphatidylcholine labeled on the choline and fatty acid moieties. Both forms were bound to albumin. Incorporation in brain lipid classes was followed within 1 h. The brain uptake of the unesterified fatty acids reached a plateau at 5-15 min and was maximal for arachidonic acid (0.45% of the perfused dose). The brain uptake of palmitoyl-lysophosphatidylcholine was similar to that of palmitic acid, whereas that of other lysophosphatidylcholines increased with the degree of unsaturation (rate and maximal uptake) and was six- to 10-fold higher than that of the corresponding unesterified fatty acid. 2-Acyl-lysophosphatidylcholines were taken up without prior hydrolysis and reacylated into doubly labeled phosphatidylcholine, which was the most labeled lipid class, whereas lipid distribution of the unesterified fatty acid was more diversified. Partial hydrolysis of 2-acyl-lysophosphatidylcholine occurred in the brain tissue, and redistribution of the fatty acyl moiety into other phospholipid classes was also observed and was the highest for arachidonic acid. In this case, the percentage of esterification of this fatty acid in phosphatidylinositol (expressed as a percentage of the total lipid fraction) was relatively lower than that observed when the unesterified form was used.(ABSTRACT TRUNCATED AT 250 WORDS)


A novel correlation between the levels of beta-amyloid protein precursor and tau transcripts in the aged human brain. beta-Amyloid protein precursor (APP) and tau are implicated in the pathogenesis of Alzheimer's disease. We quantified the levels of APP and tau transcripts in the three cortical regions of 38 aged human brains obtained from consecutive autopsied patients. The level of APP mRNA was directly proportional to that of tau mRNA to a remarkable extent, suggesting coordinate expression of the APP and tau genes, whereas much weaker correlations were noted among mRNAs encoding other neuronal proteins. From the previous data on the differential expression of APP and tau mRNAs, the levels of APP-751 and -695 mRNAs were calculated and found to be proportional to those of four-repeat and three-repeat tau mRNAs, respectively, whereas that of APP-770 mRNA was rather constant. These results suggest that the mRNA concentrations of APP isoforms are linked to those of tau isoforms in the aged human brain.


Chronic administration of lithium or other antidepressants increases levels of DARPP-32 in rat frontal cortex. We studied the chronic actions of lithium on rat brain by investigating its effects on cyclic AMP-dependent protein phosphorylation by use of a back-phosphorylation procedure. We identified one heavily regulated phosphoprotein in frontal cortex as the 32-kDa dopamine- and cyclic AMP-regulated phosphoprotein (DARPP-32). Immunoblot experiments demonstrated that chronic lithium regulation of DARPP-32 back-phosphorylation is associated with equivalent increases in levels of DARPP-32 immunoreactivity. Lithium regulation of DARPP-32 immunoreactivity required chronic drug administration and was not observed in several other brain regions examined. Moreover, chronic administration of the antidepressant imipramine or tranylcypromine produced a similar increase in levels of DARPP-32 in frontal cortex, whereas other types of psychotropic drugs, including haloperidol, morphine, and cocaine, did not influence DARPP-32 levels. Increased levels of DARPP-32 could reflect a common functional effect on frontal cortex of long-term exposure to lithium and some other antidepressant medications, an effect possibly related to the clinical actions of these drugs.


Iron-melanin complex in substantia nigra of parkinsonian brains: an x-ray microanalysis. Using energy-dispersive x-ray analysis on an electron microscope working in the scanning transmission electron microscopy mode equipped with a microanalysis system, we studied the subcellular distribution of trace elements in neuromelanin-containing neurons of the substantia nigra zona compacta (SNZC) of three cases of idiopathic Parkinson's disease (PD) [one with Alzheimer's disease (AD)] and of three controls, in Lewy bodies of SNZC, and in synthetic dopamine-melanin chemically charged or uncharged with Fe. Weak but significant Fe peaks similar to those of a synthetic melanin-Fe3+ complex were seen only in intraneuronal highly electron-dense neuromelanin granules of SNZC cells of PD brains, with the highest levels in a case of PD plus AD, whereas a synthetic melanin-Fe2+ complex showed much lower iron peaks, indicating that neuromelanin has higher affinity for Fe3+ than for Fe2+. No detectable Fe was seen in nonmelanized cytoplasm of SNZC neurons and in the adjacent neuropil in both PD and controls, in Lewy bodies in SNZC neurons in PD, and in synthetic dopamine-melanin uncharged with iron. These findings, demonstrating for the first time a neuromelanin-iron complex in dopaminergic SNZC neurons in PD, support the assumption that an iron-melanin interaction contributes significantly to dopaminergic neurodegeneration in PD and PD plus AD.


Gene expression of aromatic L-amino acid decarboxylase in cultured rat glial cells. Northern blot hybridization was performed to detect aromatic L-amino acid decarboxylase (AADC) mRNA in primary cultures of astrocytes and C6 glioma cells. The cDNA probe for rat AADC was generated by reverse transcription from rat adrenal gland total RNA and was amplified by the polymerase chain reaction method. AADC mRNA from cultured astrocytes and C6 glioma cells was present as a single band, 2.2 kbp in size, that comigrated with the RNA from rat kidney. Western immunoblot showed a single protein band at 52 kDa for AADC enzyme protein. These findings demonstrate that AADC is expressed in rat glial cells.


Quantitation of rat dopamine transporter mRNA: effects of cocaine treatment and withdrawal. Dopamine transporter mRNA levels in the rat substantia nigra were quantified using a sensitive nuclease protection assay with a highly homologous human dopamine transporter cDNA clone. The same probe was also used to visualize dopamine transporter mRNA in the substantia nigra by in situ hybridization. Repeated cocaine administration (15 mg/kg, twice a day for 6.5 days) resulted in a greater than 40% decrease in nigral dopamine transporter mRNA levels. In contrast, dopamine transporter mRNA levels were unchanged after either acute treatment (4 h before death) or repeated cocaine treatment followed by a 72-h withdrawal period. Thus, blockade of the dopamine transporter by repeated cocaine administration may result in the down-regulation of dopamine transporter gene expression in dopamine neurons.


Interleukin 1 and tumor necrosis factor-alpha stimulate the production of colony-stimulating factor 1 by murine astrocytes. Astrocytes have the ability to secrete colony-stimulating factor 1 (CSF-1), a growth factor known to stimulate the proliferation of brain macrophages. We have studied the effect of cytokines such as interleukin 1 (IL-1), tumor necrosis factor-alpha (TNF alpha), and interleukin 6 (IL-6) on the production of CSF-1 by cultured primary astrocytes and an astrocytic cell line derived from embryonic mouse brain. We observed that both TNF alpha and IL-1 increased CSF-1 mRNA and protein levels in the astrocytic cultures. In contrast, IL-6 was ineffective. The CSF-1 mRNA levels were strongly reduced by incubating immortalized astrocytic cells with staurosporine, a protein kinase C inhibitor, both in the absence and in the presence of cytokines. Conversely, 12-O-tetradecanoylphorbol 13-acetate, a protein kinase C activator, increased CSF-1 mRNA levels. These results suggest a mechanism whereby mononuclear phagocytes could favor their own recruitment in the CNS by producing cytokines.


Characterization of soluble neural cell adhesion molecule in rat brain, CSF, and plasma. The polypeptide composition and glycosylation of soluble isoforms of neural cell adhesion molecule (NCAM) in developing rat brain, CSF, and plasma were characterized. Soluble NCAM in rat brain consisted of several glycosylated isoforms. The degree of glycosylation was developmentally regulated. After desialylation, four polypeptides of M(r) values of approximately 190,000 (s1), 135,000 (s2), 115,000 (s3), and 110,000 (s4) were observed. Polypeptides s1, s2, and s3 were also present in CSF, whereas only s3 and s4 were observed in plasma. Treatment of soluble brain NCAM with N-glycosidase F, which removes N-linked carbohydrates, produced polypeptides of M(r) values of approximately 190,000, 125,000, and 108,000-97,000. The monoclonal antibody OB11, which recognizes an epitope on the cytoplasmic part of transmembrane forms of NCAM, did not react with any of the soluble isoforms. Purified soluble NCAM, consisting mainly of s3, contained an N-terminal sequence identical to that of membrane-associated NCAM. Gel filtration of s3 indicated that it was present as a dimer under the chosen conditions. NCAM-expressing glioma cells adhered specifically to immobilized soluble NCAM. This implies that functionally significant soluble forms of NCAM are present in the extracellular fluid.


Characterization of nicotinic receptor-mediated [3H]dopamine release from synaptosomes prepared from mouse striatum. This study establishes that presynaptic nicotinic receptors modulate dopamine release in the mouse striatum. Nicotinic agonists elicit a dose-dependent increase in the release of [3H]dopamine from synaptosomes prepared from mouse striatum. At low concentrations, this release is Ca2+ dependent, whereas at higher concentrations Ca(2+)-independent, mecamylamine-insensitive release was also observed. The Ca(2+)-dependent nicotine-evoked release was not blocked by alpha-bungarotoxin but was effectively blocked by neuronal bungarotoxin as well as several other nicotinic receptor antagonists. The relationship between potency for stimulation of release for agonists and potency for inhibition of release for antagonists was compared to the affinity of these compounds for the [3H]nicotine binding site. The overall correlation between release and binding potency was not high, but the drugs may be classified into separate groups, each of which has a high correlation with binding. This finding suggests either that more than one nicotinic receptor regulates dopamine release or that not all agonists interact with the same receptor in an identical fashion.


Determination of regional rates of cerebral protein synthesis adjusted for regional differences in recycling of leucine derived from protein degradation into the precursor pool in conscious adult rats. The quantitative autoradiographic L-[1-14C]leucine method for the determination of regional rates of cerebral protein synthesis in vivo takes into account recycling of unlabeled leucine derived from protein degradation into the precursor pool for protein synthesis. We have evaluated the degree of recycling by measuring the ratio of the apparent steady-state leucine specific activity in the precursor amino acid pool (tRNA-bound leucine) to that in the arterial plasma. In the whole brain of the conscious rat this ratio (lambda WB) equals 0.58. The equivalent ratio for leucine in the acid-soluble pool in whole brain (psi WB) is 0.49. A first-degree polynomial equation for lambda WB as a function of psi WB was fitted from paired determinations. To determine the degree of recycling in local regions of the brain, we have measured in individual brain regions (i) psi i and calculated lambda i assuming that the fitted equation also applies to these localized regions. Our results indicate that the degree of recycling into the precursor pool does vary regionally; lambda i in the individual regions varies from 0.62 in the hypoglossal nucleus to 0.50 in the globus pallidus. Local rates of protein synthesis were then determined by the autoradiographic technique with regional corrections for recycling of unlabeled leucine. Rates of leucine incorporation into protein averaged 6.1 nmol/g of tissue/min in the brain as a whole, with the rates in gray matter about twice those in white matter.


Cleavage of bovine brain microtubule-associated protein-2 by human immunodeficiency virus proteinase. The high-molecular-weight dendritic cytoskeletal protein known as microtubule-associated protein (MAP)-2 displays the capacity to stimulate tubulin polymerization and to associate with microtubules. Serine proteases cleave MAP-2 into a C-terminal M(r) 28,000-35,000 microtubule-binding fragment and a larger N-terminal M(r) 240,000 projection-arm region. We now show that human immunodeficiency virus (HIV) proteinase also progressively degrades purified MAP-2 in vitro. This proteolysis reaction is characterized by transient accumulation of at least six intermediates, and most abundant of these is an M(r) 72,000 species that retains the ability to associate with taxol-stabilized microtubules. Treatment of this M(r) 72,000 species with thrombin releases the same M(r) 28,000 component as that derived from thrombin action on intact high-molecular-weight MAP-2, indicating that the viral aspartoproteinase action preferentially occurs further toward the N-terminus. The association of the M(r) 72,000 component with microtubules can be disrupted by the presence of a 21-amino acid peptide analogue of the second repeated sequence in the MAP-2 microtubule-binding region. We also studied HIV proteinase action on MAP-2 in the presence of tubulin and other MAPs that recycle with tubulin, and contrary to other published studies we found no effect of such treatment on microtubule self-assembly behavior. Cleavage of isolated MAP-2 by the HIV enzyme at high salt concentrations, followed by desalting and addition of tubulin, also resulted in microtubule assembly, albeit with slightly reduced efficiency.


Endogenous noradrenaline and dopamine in nerve terminals of the hippocampus: differences in levels and release kinetics. The presence and release of endogenous catecholamines in rat and guinea pig hippocampal nerve terminals was studied by fluorimetric HPLC analysis. In isolated nerve terminals (synaptosomes) the levels and breakdown of endogenous catecholamines were determined and the release process was characterized with respect to its kinetics and Ca2+ and ATP dependence. Endogenous noradrenaline and dopamine, but not adrenaline, were detected in isolated hippocampal nerve terminals. For dopamine both the levels and the amounts released were more than 100-fold lower than those for noradrenaline. In suspension, released endogenous catecholamines were rapidly broken down. This could effectively be blocked by monoamine oxidase inhibitors, Ca(2+)-free conditions, and glutathione. The release of both noradrenaline and dopamine was highly Ca2+ and ATP dependent. Marked differences were observed in the kinetics of release between the two catecholamines. Noradrenaline showed an initial burst of release within 10 s after K+ depolarization. The release of noradrenaline was terminated after approximately 3 min of K+ depolarization. In contrast, dopamine release was more gradual, without an initial burst and without clear termination of release within 5 min. It is concluded that both catecholamines are present in nerve terminals in the rat hippocampus and that their release from (isolated) nerve terminals is exocytotic. The characteristics of noradrenaline release show several similarities with those of other classical transmitters, whereas dopamine release characteristics resemble those of neuropeptide release in the hippocampus but not those of dopamine release in other brain areas. It is hypothesized that in the hippocampus dopamine is released from large, dense-cored vesicles, probably colocalized with neuropeptides.


Effects of hypoxia on choline exchange among organs. Cerebral blood flow (CBF) and the arteriovenous (A-V) difference for choline (Ch) across brain, lung, splanchnic territory, liver, kidney, and lower limb were studied in anesthetized, mechanically ventilated rats subjected to 10-20-min periods of hypoxia induced by lowering the inspired O2 concentration to 13%. A large, time-dependent increase in arterial blood Ch concentration occurred during hypoxia. This phenomenon coincided with a net rate of uptake of Ch by the brain during hypoxia (0.81 +/- 0.24 nmol/min, n = 10; p less than 0.05), which contrasted with a net rate of loss of Ch by this organ during the control period that preceded hypoxia (-0.20 +/- 0.08 nmol/min, n = 10; p less than 0.05). During hypoxia, lungs and splanchnic territory showed negative A-V differences for Ch levels (net Ch loss), whereas brain, liver, kidney, and lower limb showed positive A-V differences for Ch levels (net Ch uptake). Ch output from lungs was already detected at 5 min within the period of hypoxia and reversed rapidly after restoration of normal oxygenation. On the other hand, Ch output from the splanchnic territory became evident only 10 min after commencement of hypoxia and outlasted this experimental condition. It is concluded that extracerebral production of Ch during hypocapnic hypoxia raises the arterial concentration of this molecule and, by reversing the gradient across cerebral capillaries, prevents the cerebral loss of Ch in this condition.


Effect of nitroprusside (nitric oxide) on endogenous dopamine release from rat striatal slices. It is becoming apparent that the synthesis of nitric oxide (NO) from L-arginine not only explains endothelium-dependent vascular relaxation, but is a widespread mechanism for the regulation of cell function and communication. We examined the role of NO on the endogenous dopamine (DA) release from rat striatum. Nitroprusside, in the concentration range of 3-100 microM, induced a dose-dependent increase in the endogenous DA release from rat striatal slices. The maximal response was 330% over the baseline release. A higher concentration of nitroprusside (300 microM) produced an inhibitory effect on the spontaneous release of DA. L-Arginine (10 and 100 microM), a substrate in the NO-forming enzyme system, also produced an elevation of DA release. L-Arginine-induced DA release was attenuated by NG-monomethyl-L-arginine, an inhibitor of NO synthase. NADPH (1 microM), a cofactor of NO synthase, enhanced L-arginine-induced DA release. These results suggest a possible involvement of NO in the DA release process in rat striatum.


Cytokine regulation of nerve growth factor-mediated cholinergic neurotrophic activity synthesized by astrocytes and fibroblasts. The neurotrophic activity of astrocytes and fibroblasts and its regulation by various cytokines were investigated. Astrocyte conditioned medium (ACM) enhanced the survival of neurons and the proliferation of astrocytes in embryonic cortical cultures grown in serum-free defined medium. However, these results were not affected by acidic fibroblast growth factor, interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF alpha), and transforming growth factor-beta 1. In contrast, ACM induced choline acetyltransferase expression in septal cholinergic neurons via nerve growth factor (NGF)-dependent and -independent mechanisms. However, neither acidic nor basic fibroblast growth factor is involved in this biological activity in ACM. The cytokines listed above mainly stimulate NGF-mediated cholinergic neurotrophic activity in ACM. A combination of IL-1 beta and TNF alpha significantly enhanced choline acetyltransferase activity in septal neurons co-cultured with astrocytes, and this effect was found to be mediated by NGF produced by activated astrocytes. Effects of astrocytes on GABAergic neurons were also examined. ACM was found to increase glutamate decarboxylase activity in neuronal cultures from septum in the presence of Ara-C. However, the cytokines did not enhance this activity in ACM. Moreover, a combination of IL-1 beta and TNF alpha had no effect on glutamate decarboxylase activity in septal neurons co-cultured with astrocytes. In a final set of experiments, cholinergic neurotrophic activity in skin-derived fibroblast conditioned medium (FCM) was examined. FCM was found to possess biological activity similar to that of ACM on septal neurons grown in serum-free defined medium with Ara-C. The cytokines also enhanced NGF-mediated cholinergic neurotrophic activity in FCM. Astrocytes and fibroblasts were found to possess NGF-type and non-NGF-type cholinergic neurotrophic activity, and various cytokines were found to regulate the NGF-type cholinergic neurotrophic activity in both types of cells. NGF produced by astrocytes and fibroblasts that are activated by cytokines is likely to be important for development and regeneration of NGF-sensitive neurons in the central and peripheral nervous systems.


Local blood-brain barrier in the newborn rabbit: postnatal changes in alpha-aminoisobutyric acid transfer within medulla, cortex, and selected brain areas. Postnatal changes in local permeability of the blood-brain barrier to an inert neutral amino acid (alpha-[14C]-aminoisobutyric acid) were investigated in 25 rabbits. The local transfer constant (K) for this tracer was measured with quantitative autoradiographic techniques at postnatal ages of 1, 3, 8, and 17 days, and adult. In adults, the amino acid penetrated the blood-brain barrier poorly in most regions examined (K less than 1 microliter.g-1.min-1) except within and in proximity to structures with a relatively leaky blood-brain barrier such as area postrema and choroid plexus. The rate of tracer entry into "impermeable" regions was seven- to 10-fold greater in 1-day-old rabbits than adults and not dependent on active transport. In young animals, there was a pronounced regional variation in K with the lowest values occurring in white matter and the highest in gray matter such as cerebral cortex, posterior thalamus, and hippocampus. During postnatal development, K decreased (p less than 0.01) with most regions having values near those of adults by 17 days of age. The results indicate that the blood-brain barrier of the newborn rabbit is relatively leaky to a small hydrophilic nonelectrolyte with a distribution that is heterogeneous regionally. Irrespective of age, such blood-borne substances can accumulate in certain brain areas considered to have impermeable vessels (e.g., nucleus tractus solitarii).


Can people hear the pitch change on a variable-pitch pulse oximeter? The introduction of the variable-pitch feature on pulse oximeters in 1983 by the Nellcor Corporation (Hayward, CA) allowed users to rapidly detect changes in oxygen saturation by listening for changes in the pitch of the tones emitted by the pulse oximeter. A few individuals have reported that they have been unable to detect a change in pitch when oxygen saturation changes. To these individuals, the variable-pitch feature of these pulse oximeters has not been beneficial. Using the pitches from one manufacturer of oximeters, we created a computer program to simulate the pitches that accompanied various oxygen saturations. The pitches were recorded onto a tape player and played for 75 volunteer subjects unfamiliar with the pitches of a variable-pitch pulse oximeter. Of our sample, 67% were able to detect a single change in pitch corresponding to a 1% fall in oxygen saturation, and 11% of the population could not detect a change in pitch until there was a change in pitch with every beat. We suggested four alternative designs that may prove beneficial to this group of individuals.


Microdetermination of 2-deoxyglucose and 2-deoxyglucose 6-phosphate to determine glucose utilization rates in single neurons and small CNS regions after injecting nontracer amounts of 2-deoxyglucose. A nontracer amount (0.25 mmol/kg of body weight) of 2-deoxyglucose (DG) was intravenously injected into rats, which were frozen 2 and 4 min later in liquid nitrogen. Freeze-dried samples of CNS regions and cell bodies of spinal motor neurons were prepared, and the concentrations of glucose, glucose 6-phosphate, DG, and DG 6-phosphate (DG6P) in them were microassayed after 3,000-1,500,000-fold amplification using an enzymatic amplification reaction, NADP cycling. Based on the time course of glucose, DG, and DG6P concentrations in arterial plasma and the anterior horn of the spinal cord, the Sokoloff-type rate equations for DG and DG6P concentrations were mathematically solved, and the resultant DG and DG6P concentration functions were fitted to the data points using the nonlinear least-squares fitting SALS package program. This fitting provided four rate constants for the functions and supported the theoretical basis for our calculations of glucose utilization rate (GUR) when DG was administered in nontracer amounts. The GUR was highest in the spinal motor neurons and lowest in the white matter of the cerebellum. Neuron-rich structures, such as the cerebellar molecular and granular layers and the anterior horn of the spinal cord, had higher GUR values than the white matter of the cerebellum and spinal cord.


Ouabain-sensitive choline transport system in capillaries isolated from bovine brain. In physiological conditions, there is a net transport of choline from brain to blood, despite the fact that the choline concentration is higher in plasma than in CSF. Because of the blood-brain barrier characteristics, such passage against the concentration gradient takes place necessarily through endothelial cells. To get a better understanding of this phenomenon, [3H]choline uptake properties have been analyzed in capillaries isolated from bovine brain. [3H]Choline uptake was linear with time for up to 1 h. Nonlinear regression analysis of the uptake rates at different substrate concentrations gave the best fit to a system of two components, one of which was saturable (Km = 17.8 +/- 4.8 microM; Vmax = 11.3 +/- 3.4 pmol/min/mg of protein) and the other of which was nonsaturable at concentrations up to 200 microM. The [3H]choline transport was significantly reduced in the absence of sodium and after incubation with 10(-4) M ouabain for 30 min. Ouabain also inhibited choline uptake in purified cerebral endothelial cells, but not in the endothelium isolated from bovine aorta. Accordingly, cerebral endothelial cells were able to concentrate [3H]choline, with this effect being abolished by ouabain, whereas in aortic endothelial cells the [3H]choline intracellular concentration was never higher than that of the incubation medium. These results suggest that the blood-brain barrier endothelium is specifically provided with an energy-dependent choline transport system, which may explain the choline efflux from the brain and the maintenance of a low choline concentration in the cerebral extracellular space.


Effects of [Sar1]angiotensin II on proenkephalin gene expression and secretion of [Met5]enkephalin in bovine adrenal medullary chromaffin cells. We have studied the effect of [Sar1]angiotensin II [S1-AII; a degradation-resistant analogue of angiotensin II (AII) on the release of [Met5]enkephalin (ME) and proenkephalin A (proENK) gene expression. Short-term (15-min to 1-h) stimulation of bovine adrenal medullary chromaffin (BAMC) cells with S1-AII at concentrations from 0.1 to 100 nM had no significant effect on secretion of ME, whereas high concentrations of S1-AII (3 to 100 microM) produced a concentration-dependent increase in the concentration of ME in the incubation media. In contrast, long-term (3- to 24-h) stimulation with low concentrations (0.1 nM-1 microM) of S1-AII increased the secretion of ME in a concentration-dependent manner (EC50 = 1 nM). The intracellular level of ME was not changed by long-term treatment with S1-AII (100 nM). In addition to increased ME secretion, long-term (24-h) stimulation with S1-AII increased the expression of proENK mRNA in a concentration-dependent manner (EC50 = 4 nM). Losartan (2-n-butyl-4 chloro-5-hydroxymethyl-1-[(2'-(1 H-tetrazol-5-yl)biphenyl-4-yl)- methyl]imidazole potassium salt, a type 1 AII receptor antagonist) inhibited these effects, whereas PD123319 (50 microM, a type 2 AII receptor antagonist) was inactive. Our results suggest that AII in BAMC cells exerts a major effect on the long-term regulation of expression of proENK mRNA and secretion of ME. These effects appear to be mediated by type 1-like AII receptors.


Dihydroxyphenylalanine and dopamine are released from portal vein together with noradrenaline and dihydroxyphenylglycol during nerve stimulation. The overflows of 3,4-dihydroxyphenylalanine, dopamine, noradrenaline, and 3,4-dihydroxyphenylglycol in canine portal vein superfused in vitro were studied before, during, and after depolarization of sympathetic nerve endings. The four compounds were separated from superfusate and from tissue on Sep-Pak C-18 cartridges and quantified by HPLC with electrochemical detection. Physiological and biochemical methods were used to show that the compound released was most probably 3,4-dihydroxyphenylalanine; the identity of the other endogenous compounds has been established previously. Release of 3,4-dihydroxyphenylalanine was calcium and frequency dependent, inhibited by a-m-L-p-tyrosine (an inhibitor of tyrosine hydroxylase) and augmented by 3-hydroxybenzylhydrazine (an inhibitor of aromatic amino acid decarboxylase). The overflows of dopamine, noradrenaline, and 3,4-dihydroxyphenylglycol from the vein were calcium and frequency dependent. It was estimated that under control conditions, approximately 80% of the total 3,4-dihydroxyphenylalanine that was synthesized was directed to catecholamine biosynthesis, approximately 8% overflowed from the vein, and approximately 14% remained unchanged within the tissue. It is concluded that 3,4-dihydroxyphenylalanine and dopamine are released together with noradrenaline and 3,4-dihydroxyphenylglycol from portal vein upon nerve depolarization.


Radial artery cannulation: a comparison of 15.2- and 4.45-cm catheters. Eighty-nine patients were studied prospectively to compare the incidence of postdecannulation arterial thrombosis and ischemic complications associated with percutaneous insertion of two different radial artery catheters. Patients scheduled for peripheral vascular surgery were randomized to receive a 15.2-cm (6 in, Argon Medical Corp.) or 4.45-cm (1.75 in, Arrow International, Inc.) 20-gauge, Teflon catheter. Extremity blood flow was evaluated prior to cannulation and again after decannulation with the modified Allen's test, pulse-volume plethysmography, and Doppler ultrasound. The incidence of postdecannulation radial artery occlusion for 15.2-cm catheters was significantly less than for 4.45-cm catheters (4 of 45 cases versus 11 of 44 cases, p = 0.05). No case of temporary or permanent ischemic injury occurred. Radial artery transfixion (16 of 45 cases versus 5 of 44 cases, p = 0.01) and hematoma formation (5 of 45 cases versus 0 of 44 cases, p = 0.02) occurred more frequently during insertion of 15.2-cm catheters than 4.45-cm catheters. The number of arterial punctures during catheter insertion and the duration of cannulation were similar for both groups. Of the 8 patients with positive modified Allen's test who underwent radial artery cannulation, one suffered arterial occlusion. Radial artery cannulation with a 15.2-cm catheter was associated with a lower incidence of postdecannulation radial artery thrombosis than cannulation with the 4.45-cm catheter. Radial artery cannulation with longer catheters (greater than 5.0 cm) appears to be a safe practice.


Educational computer simulation of malignant hyperthermia. An educational graphic simulator was developed to provide an interactive learning environment to practice the diagnosis and treatment of malignant hyperthermia. The program incorporates a set of dynamically interacting models to present the physiologic changes associated with malignant hyperthermia and the simulated patient's response to management. Cardiovascular, respiratory, and temperature changes are presented through a graphic display of the operating room monitors. Mouse-driven input is used to manage the airway, control ventilation, manage cardiovascular and rhythm disturbances, and control fluids, electrolytes, and temperature. Medications, including dantrolene, antidysrhythmics, diuretics, and sodium bicarbonate, can be administered. Four simulated patients with different presentations of malignant hyperthermia are included to illustrate variations in the syndrome. Two of these patients are described in detail.


Pulmonary artery catheter monitoring during single-lung ventilation in dogs. The effect of pulmonary artery (PA) catheter insertion on cardiac output measurements and the effect of single-lung ventilation on data derived from catheters placed in both the right and left pulmonary arteries were examined in 17 anesthetized mongrel dogs. Serial tricarboncyanine dye measurements of cardiac output were taken at baseline and after insertion of each of two PA catheters, one catheter in each lung, to detect possible changes in cardiac output due to instrumentation. In the second part of the study, single-lung ventilation was achieved through a Kottmeier double-lumen tube. Right and left paired values of thermodilution cardiac output, PA systolic pressure, and PA PO2 were compared during ventilation of both lungs, during right lung ventilation, during return to ventilation of both lungs, and during left lung ventilation. Mean dye-dilution cardiac outputs were not significantly different after one or two catheters were inserted. Paired right and left values were similar for thermodilution cardiac output, PA PO2, and pulmonary systolic pressure, regardless of ventilation conditions. Even during single-lung ventilation, data obtained from PA catheters in the ipsilateral and contralateral lung were equivalent in supine dogs.


Correlated, simultaneous, multiple-wavelength optical monitoring in vivo of localized cerebrocortical NADH and brain microvessel hemoglobin oxygen saturation. Current forms of brain monitoring, such as electroencephalography (EEG), have had limited clinical utility. The EEG records spontaneous cerebrocortical activity and thus is an indirect indicator of metabolic demand and, to a lesser extent, an indicator of mismatch of supply versus demand. Ischemia modulates EEG activity in ways that can usually be detected, but EEG patterns can be similarly modulated by many other factors, including temperature and pharmacologic manipulation. This in vivo study in physiologically monitored animals evaluated the use of correlated optical spectroscopy, performed with an instrument having a fiberoptic light-guide bundle in contact with the cerebral cortex, for the simultaneous monitoring of cerebrovascular oxygen availability and intracellular oxygen delivery. A highly specific monitor of cerebral intracellular oxygen supply, the cerebrocortical intramitochondrial NADH redox state, was monitored in vivo with a fluorescence technique. Absorption spectroscopy was used concurrently to monitor hemoglobin content (blood volume) and oxygen saturation in the microcirculation. Correlated changes in optical signals from cerebrocortical NADH and hemoglobin were studied in a swine model (n = 7) of nitrogen hypoxia. Measurements were made at four wavelengths with a time-division, multiplexed fluorometer/reflectometer. Because the NADH fluorescence signal at 450 nm is affected by local changes in blood volume, a "corrected" fluorescence signal is usually calculated. In previous studies, where only two wave lengths have been measured, attempts at correction were based on reflectance at the excitation wavelength (366 nm). We compared estimators of changes in microcirculatory blood volume using reflection at two wavelengths: 366 nm and 585 nm, the wavelengths for maximum and isobestic absorption. The results of the studies were as follows: (1) during transient hypoxia, NADH and local hemoglobin saturation signals changed in concert with arterial pulse oximetry, with changes in NADH lagging behind changes in saturation by an average of 5.3 seconds; (2) after hypocapnic ventilation to a mean PaCO2 of 20.2 +/- 0.8 mm Hg, NADH increased by 11.5 +/- 8.7% (as compared with maximal change during anoxia), local hemoglobin saturation decreased by 7.7 +/- 6.4%, and local blood volume decreased by 12.5 +/- 13%, while arterial SpO2 was unchanged; (3) our two measures of local blood volume were closely correlated during carbon dioxide perturbations, but poorly correlated during hypoxic perturbation; and (4) NADH fluorescence provided a more rapid, sensitive indicator of oxygen deprivation than did the EEG. During transient hypoxia, EEG changes occurred 57.4 +/- 10.4 seconds after the onset of decline in local hemoglobin saturation, after NADH had completed 50% of its maximal increase.


Use of end-tidal PCO2 and transcutaneous PCO2 as noninvasive measurement of arterial PCO2 in extubated patients recovering from general anesthesia. This study was designed to assess the accuracy of end-tidal PCO2 and transcutaneous PCO2 as measurements of arterial PCO2 in extubated, spontaneously breathing patients recovering from general anesthesia. In 30 patients, measurement of arterial transcutaneous, and end-tidal PCO2 were taken simultaneously with body temperature approximately every 15 minutes over a 2-hour period. Arterial PCO2 values were corrected for body temperature. Values for PaCO2 were compared with those for PETCO2 and PsCO2 by linear regression analysis and by calculation of bias +/- precision. Thirty-six percent of the capnogram tracings obtained did not develop a plateau phase. We found poor correlation between end-tidal and arterial PCO2 regardless of the shape of the capnogram tracing, as well as poor correlation between transcutaneous and arterial PCO2. Although the measurements of bias and precision of noninvasive PCO2 monitors in this population are comparable to studies in other populations, we advise caution in relying on the routine use of PETCO2 or PsCO2 for the noninvasive assessment of respiratory depression in extubated, spontaneously breathing patients recovering from general anesthesia.


Inconsistencies in the correlation between loss of brain stem auditory evoked response waves and postoperative deafness. This case underscores the difficulty of predicting postoperative hearing status from brain stem auditory evoked response (BAER) monitoring when wave I is preserved and all later waves are lost. During an operation involving the base of the skull, sudden and irreversible loss of all BAER waves beyond wave I occurred unilaterally. Wave I was preserved, with reduced amplitude and minimal latency shift. There was no permanent postoperative hearing sensitivity loss or speech discrimination loss.


Capnography does not reliably detect double-lumen endotracheal tube malplacement. Two patients are described in whom double-lumen endotracheal tube malplacement and its ventilatory consequences were not detected by infrared capnography. Problems were suspected on auscultation, and the malplacement was diagnosed by means of bronchospirometry. We conclude that bronchospirometry helps detect problems with endotracheal intubation.


Decline of vasopressin immunoreactivity and mRNA levels in the bed nucleus of the stria terminalis following castration. Vasopressinergic (VP) neurons in the bed nucleus of the stria terminalis (BNST) of the rat are regulated by gonadal steroids. Gonadectomy causes the projections of the BNST to lose their VP immunoreactivity gradually over a period lasting more than 2 months. Here we have compared the rate of decline of VP mRNA and VP immunoreactivity in the BNST of adult male rats following castration. In experiment 1, the peak number of VP-immunoreactive cells and the level of VP gene expression were compared in sham-operated controls and at 1, 3, or 8 weeks postcastration. The number of VP-immunoreactive cells was not decreased at 1 week postcastration but was significantly reduced (p less than 0.0001) at 3 and 8 weeks postcastration. VP gene expression declined more rapidly, and both the total number of labeled cells (p less than 0.0001) and the average number of grains per cell (p less than 0.01) were significantly reduced by 1 week postcastration. No VP-expressing cells were detectable at 3 or 8 weeks. The difference in the rate of decline in the number of cells labeled by the two techniques following castration did not appear to be due to colchicine pretreatment. In experiment 2, VP mRNA in the BNST was compared in sham-operated controls or at 1, 3, or 7 d postcastration. A significant decrease (p less than 0.01) in the average number of grains per cell was detectable by just 1 d following castration, and the number of labeled cells was significantly reduced (p less than 0.001) by 3 d postcastration.(ABSTRACT TRUNCATED AT 250 WORDS)


Synaptic interactions between GABA-immunoreactive profiles and the terminals of functionally defined myelinated nociceptors in the monkey and cat spinal cord. This study analyzes the synaptic interactions between the central terminals of A delta high threshold mechanoreceptors (A delta HTMs) and GABA-immunoreactive profiles. A delta HTM primary afferents from three monkeys and one cat were electrophysiologically identified and intracellularly labeled with HRP, and their terminal arborizations in laminae I and II of the sacrocaudal spinal cord were studied at the ultrastructural level. GABA-immunoreactive profiles in relation to A delta HTM terminals were demonstrated using postembedding colloidal gold techniques. Monkey A delta HTM terminals (n = 131) usually constituted the central element of synaptic glomeruli; they established large asymmetric synaptic contacts with 1-13 dendrites (modal value 2-4) and were surrounded by 0-6 peripheral axon terminals (modal value 2-3). The large majority (around 85%) of the peripheral axon terminals were GABA immunoreactive. They were found presynaptic to the A delta HTM terminal and/or to dendrites postsynaptic to the primary afferent terminal. Furthermore, all peripheral axon terminals found presynaptic to the A delta HTM terminals showed GABA immunoreactivity. Within a single A delta HTM fiber, this synaptic arrangement was found in 20-60% of its boutons. In addition, 28% of the postsynaptic dendritic profiles displayed weak GABA immunoreactivity. Some of them contained vesicles; however, only in a few cases did we observe synapses between a GABA-immunoreactive vesicle-containing dendrite and a dendritic profile postsynaptic to an A delta HTM terminal. Similar synaptology and interactions with GABA-immunoreactive profiles were displayed by the terminals of the single cat A delta HTM fiber studied. Our data support the hypothesis that GABA-containing neurons use both presynaptic and/or postsynaptic mechanisms to exert a powerful control, presumably inhibitory, over the transmission of nociceptive information between A delta HTM afferents and second-order neurons in monkey and cat spinal cord. Our results also imply that GABA may be released within the synaptic glomeruli formed by A delta HTM terminals either by local dendrites or by axon terminals. We discuss the possibility that these GABAergic synapses can be driven by inputs from both primary afferents and/or descending systems to modulate the transmission of nociceptive sensory information.


Neurophysiological correlates of hand preference in primary motor cortex of adult squirrel monkeys. Variability in the functional topography of area 4 was examined in adult squirrel monkeys. Conventional intracortical microstimulation techniques were used to derive detailed maps (250 microns interpenetration distances) of distal forelimb movement representations in both hemispheres of six monkeys. Spatial features of these representational maps were then compared to the hand preferred by the individual animals during a motor task requiring skilled digit use. Beyond a few broad generalizations common to all area 4 motor maps, the local mosaic-like topography of individual distal forelimb representations was highly idiosyncratic. Using statistical procedures to determine the independent contributions of individual, side, and movement category to the total variation in motor maps, the results demonstrate statistically significant variation in representational topography among individuals as well as between hemispheres of the same individuals. In the dominant hemisphere (i.e., the hemisphere opposite the preferred hand), the distal forelimb representations generally were greater in number and larger in total area, and displayed a longer total boundary length and a greater index of spatial complexity. Because of the direct relationship between interhemispheric asymmetry and behavioral asymmetry, these studies suggest that a large source of variability found in the topography of motor maps in this and other studies derives from differences in the way particular movements and/or movement combinations are performed by individual animals.


Segregation of presynaptic inputs on an identified target neuron in vitro: structural remodeling visualized over time. Sensory cells of Aplysia form chemical synapses with the motor cell L7 in culture. Under certain conditions, sensory cells will also form electrical connections with each other. Sites of chemical synaptic interaction between the sensory cells and L7 are located at varicosities along sensory cell processes that overlie the main axons of L7, since these structures have been shown ultrastructurally to contain active zones. Previous studies have suggested that the distribution of sensory cell varicosities can be restricted to exclusive regions of the motor cell by the presence of other sensory cells. We wished to investigate (1) how this segregated pattern is generated over time and (2) whether electrical coupling between sensory cells has an effect on this segregated pattern. Using fluorescent dye injection and low-light video microscopy, we visualized the distribution of varicosities for each of two sensory cells growing on L7. In cases in which sensory cells are not electrically coupled, the varicosities from these two cells are spatially segregated on the target after 4 d in culture but not after 2 d in culture. Examination of the varicosity distribution of the same sensory cells on the second and third day of growth indicated both an increased rate in the elimination of varicosities from previously occupied areas and a restriction of varicosity formation in new areas of the target when a second sensory cell is present. For sensory cells that are electrically coupled, varicosities from these cells were not spatially segregated on the target even after 4 d in culture. These observations in vitro suggest that segregation of synaptic inputs by Aplysia sensory cells, which show little spontaneous activity of action potentials, can emerge over time via a process that includes both the elimination of existing sensory varicosities and the restriction of new varicosity formation. Our results also suggest that electrical connections between presynaptic cells can disrupt the segregation of their varicosities on a target, resulting in significant changes in the developing connectivity.