Some metabolic relationships in young patients with ischemic heart disease. The interrelationships between body weight, insulin secretion and serum lipids were studied in 40 young white patients (mean age 37 years) with established ischemic heart disease (IHD), living in Johannesburg. None was severely obese, hypertensive or overtly diabetic. In general, strong positive correlations were found between body weight and insulin concentrations and between insulin levels and fasting serum triglycerides. However, insulin levels were relatively low in 4 patients with marked hypertriglyceridemia (above 350 mg/dl). These data are consistent with the postulate that insulin promotes (hepatic) triglyceride synthesis, but when there is gross hypertriglyceridemia peripheral triglyceride clearance becomes defective. Insignificant correlations were observed between body weight and serum lipids and between cholesterol and other metabolic variables. We conclude that there is a sequential link between increasing body weight, insulin secretion and triglyceride levels in young patients with IHD, but that cholesterolemia is independent of this axis.


[The effects of Triton WR 1339 on the secretion of bile lipids in the rat]. Rats bearing a biliary fistula received i.v. a solution of Triton WR 1339. Bile and plasma lipid composition was studied every 2 hrs and compared to that of control rats injected with saline. Two hours after Triton injection a sharp decrease in the bile secretion of lecithins (-- 93%), cholesterol (-- 50%) and bile salts (-- 50%) was observed together with a fall in bile flow (-- 20%). Eight hours after Triton administration the biliary output of lecithins, cholesterol and bile salts was lowered to --73%, --50% and --34% respectively compared to control animals. At that time an accumulation of L.C.A.T. substrates (lecithins and cholesterol) was observed in the plasma of Triton group. These modifications of biliary lipids after inhibition of L.C.A.T. activity by Triton W 1339 could be the result of a decreased production of plasma lysolecithins and cholesterol esters suggesting that both lipids could be important precursors for the synthesis of bile constituents. Furthermore this support the view that the production by the liver of plasma and bile lipids follows two distinct pathways.


Roentgenological and endoscopic findings of malignant lymphoma. Roentgenological and endoscopic characteristics of malignant lymphoma of the stomach were studied referring to the gross pathological appearance of the tumors. Based on the results obtained, we classified malignant lymphoma of the stomach into four types, i.e., superficial, ulcer, polypoid and giant fold types.


Duplication of aortic wall seen by echocardiography. In four patients, echocardiography showed duplication of an aortic wall echo. An aortic dissection was present in only one. Other causes for the echocardiographic pattern were abscess in the interventricular septum, mitral stenosis, and dilatation of the noncoronary sinus of Valsalva. Division of the echo from the mitral ring into two separate posterior aortic wall echoes was seen only in the patient with dissecting aneurysm; it is suggested that this appearance adds to the specificity of the finding of aortic wall duplication by echocardiography in the diagnosis of aortic dissection.


'Incessant' tachycardias in Wolff-Parkinson-White syndrome. II: Role of atypical cycle length dependency and nodal-his escape beats in initiating reciprocating tachycardias. Descriptions of patients with the Wolff-Parkinson-White (WPW) syndrome and reciprocating tachycardia in whom the initiation of the arrhythmia depended neither on the occurrence of premature beats nor on antecedent cycle-length shortening are given. In 5 the occurrence of escape beats in the bundle of His, usually in the presence of sinoatrial disease, activated the tachycardia circuit, but in the other 2 there were unusual mechanisms related to bradycardia-dependent block in the anomalous pathway, and delayed response to shortening of the atrial cycle length, respectively. Careful assessment of such mechanisms is essential for the correct choice of antiarrhythmic prophylactic therapy.


Coronary artery disease in patients dying from cardiogenic shock or congestive heart failure in the setting of acute myocardial infarction. Pathological findings in the heart and particularly in the coronary arteries are reported from 70 patients dying from pump failure after acute myocardial infarction. Fifty of the patients had died in cardiogenic shock, the remainder from refractory congestive heart failure. Three-vessel disease (greater than or equal to 75% occlusion) was present in 68 per cent of the group with cardiogenic shock but in only 35 per cent of those with fatal congestive heart failure (P less than 0-02). In both groups there was an almost equal incidence (84% for cardiogenic shock and 80% for congestive heart failure) of severe disease (greater than or equal to 75% occlusion) over a long segment of the left anterior descending artery. However, there were differences between the two groups regarding the involvement of the other coronary arteries. Whereas patients with cardiogenic shock generally showed severe disease over a long segment in all coronary arteries, in 60 per cent of those with congestive heart failure there was only local severe narrowing of the right coronary artery with little or no narrowing of the peripheral part. Similarly, 60 per cent of those with congestive heart failure had less than 75 per cent narrowing in the left circumflex artery. These anatomical findings may be of relevance with regard to desirability of acute coronary bypass surgery in patients with pump failure after acute myocardial infarction.


Diagnosis of endocardial cushion defect with cross-sectional and M-mode scanning echocardiography. Differentiation from secundum atrial septal defect. Twelve cases of endocardial cushion defect were studied before and after operation with ultrasono-cardiotomography (tomography) cross-sectional echocardiography, two-dimensional echocardiography, B-scan echocardiography) and M-mode scan along a horizontal section of the heart. For comparison, 20 healthy subjects, 18 cases of mitral valvular disease, 4 cases of congestive cardiomyopathy, 1 case of partial anomalous pulmonary venous drainage, and 25 cases of atrial septal defect of secundum type were also examined with the same technique. In cases without cardiac malformation, the echo of the anterior mitral valve was usually continuous medially with that of the interatrial septum in the horizontal plane at the level of the membranous septum. This feature was clearly recorded in all cases with right heart enlargement. In ostium secundum atrial septal defect the echo of the anterior mitral valve continued into that of the interatrial septum. An echo interruption was shown, indicating the defect itself to be in the middle part of the interatrial septum. In all the cases of endocardial cushion defect which we examined discontinuity was shown between the echo of the anterior mitral valve and that of the interatrial septum. This discontinuity was interpreted as indicating the defect itself. The mitral valve ring echo was close to the basal end of that of the interventricular septum, possibly reflecting an abnormal attachment of the mitral valve. In all cases, after operation, the echo of the artificial interatrial septum was recorded, continuous with that of the anterior mitral valve. The features of the echocardiographic sweep from the anterior mitral valve to the interatrial septum were thus different in the three groups. These echocardiographic differences are thought to correspond to the anatomical differences between the normal, atrial septal defect of secundum type, and endocardial cushion defect, and are essential features differentiating them from each other.


Familial sinoatrial node dysfunction. Increased vagal tone a possible aetiology. Clinical and electrophysiological studies of a 13-year-old boy with sinus bradycardia revealed sinus node dysfunction. Long-term follow-up data of members of his family indicated familial sinus node dysfucntion. Increased vagal tone was present in all patients. It is suggested that excessive vagal discharge for a pronlonged time may be the basic mechanism of sinus node dysfunction in these patients.


Surgical treatment of pulmonary atresia with intact ventricular septum. Fifty-nine children with pulmonary atresia and intact ventricular septum underwent various forms of surgical treatment at the Hospital for Sick Children, Toronto, during 1950 to mid 1975. Twenty-three patients had pulmonary valvotomy, 15 direct, 2 indirect, and 6 both direct valvotomy and infundibulectomy. All died, 19 early and 4 late. Of 13 patients who received a systemic-pulmonary artery shunt, 4 combined with surgical atrial septectomy, there are only 2 long-term survivors both of whom were children who had had a Waterston anastomosis. Recently we have been treating infants with small right ventricles with balloon atrial septostomy at cardiac catherterization followed by a Potts anastomosis and pulmonary valvotomy. If the Potts anastomosis appears satisfactory the persistent ductus arteriosus is ligated. This scheme was used in 23 infants, with 4 early deaths and 2 late deaths. Of 17 survivors, further shunts were required in 4 children. One child has had a formal repair, with insertion of valves in both tricuspid and pulmonary areas. We believe that this operative combination of Potts anastomosis and pulmonary valvotomy offers the infant with pulmonary atresia and a small right ventricle a relatively low initial mortality and the possibility of right ventricular enlargement and subsequent repair.


The Wenckebach phenomenon between electric pacemaker and ventricle. Two cases which exhibited a Wenckebach form of exit conduction between an electrical pacemaker and the ventricular myocardium are presented. This manifests with increasing latency (stimulus to QRS  intervals) and ultimate stimulus failure (block). The cycle then repeats itself. The QRS duration also increases progressively within each cycle. The phenomenon connotes an adverse prognosis. Several postulates regarding the mechanism are discussed.


Quantitative radionuclide angiocardiography. Determination of left ventricular ejection fraction in children. A method is described for measuring left ventricular ejection fraction which uses high frequency computer recording of gamma scintillation camera data and peripheral venous injectinon of technetium-99m as sodium pertechnetate. Data from mechanical model experiments are used to show feasibility of this method. A phantom experiment is described which was used to develop a technique for accurate delineation of the ventricular outline in the presence of background. The left ventricular ejection was measured in 12 patients by radionuclide angiocardiography and biplane cineangiography. Comparison of these two methods gave a correlation coefficient of 0-91. In addition, left ventricular ejection fraction was measured in 34 patients (aged 7 weeks to 18 years) without evidence of cardiac disease using the radionuclide method alone. Average ejection fractions of 0-66 and 0-70 were found for children over 2 years of age and children 2 years of age or younger, respectively. In addition, an interobseerver comparison study was performed with the data from 10 patients, and only small differences were noted (SD 0-025).


Prevention of balloon damage during initiation of intra-aortic balloon counterpulsation. Damage to an intra-aortic balloon occurred on two occasions in 93 insertion procedures. The complication was recognized before gas embolization occurred, and neither patient suffered ill effects. On both occasions the balloon was holed when pinched by artery forceps applied to the 'dacron' graft.


Haemolysis after mitral valve replacement with the Björk-Shiley and the Lillehei-Kaster disc valve prosthesis. The present study reports on the incidence and degree of intravascular haemolysis in 33 patients with Björk-Shiley tilting disc prosthesis and in 34 patients with Lillehei-Kaster pivoting disc valve prosthesis in the mitral position examined 12 to 24 months after the operation. Serum haptoglobin, serum lactate dehydrogenase, serum bilirubin, and haemoglobin estimations were performed. Significant haemolysis was detected in 85 per cent of the patients. Haptoglobin was absent or reduced in 72 per cent, while raised values for serum lactate dehydrogenase were found in 43 per cent. The increase in lactate dehydrogenase was moderate and showed no correlation with blood flow through the prosthesis or with the gradient across the prosthesis. Most patients had normal haemoglobin and normal serum bilirubin values. Although intravascular haemolysis was of little clinical significance, the increase in lactate dehyrdrogenase was significantly higher in patients with the Lillehei-Kaster prosthesis than in patients with the Björk Shiley prosthesis, indicating a slightly shorter red cell lifespan in the former group. The possible reasons for the difference between the two groups are discussed.


Lack of effect of bed rest and cigarette smoking on development of deep venous thrombosis after myocardial infarction. In a prospective study of patients admitted to a coronary care unit, the incidence of isotopically diagnosed deep venous thrombosis was found to be related to the severity of illness rather than to the duration of bed rest. In addition, no negative correlation was found between cigarette smoking and deep venous thrombosis.


Transiet atrial fibrillation after minor head injury. Two patients developed transient rapid atrial fibrillation after a blow on the head. There was no evidence of neurological damage or organic heart disease on subsequent investigation. Neither patient was aware of the cardiac irregularity.


Evaluation of rapid atrial pacing in diagnosis of coronary artery disease. Evaluation of atrial pacing test. Seventeen patients presenting with anginal-type pain were studied by bicycle exercise testing, rapid atrial pacing, and coronary angiography. Ten patients with angina and abnormal pacing tests at rates less than 180/minute were found to have significant coronary artery disease as demonstrated by coronary angiography. Seven patients with pacing-induced chest pain only at rates of 180 and above had normal coronary angiogram. This suggests that patients requiring rates of 180 or more to produce a positive atrial pacing test, following our protocol, do not usually have significant coronary artery disease though confirmation requires a larger study.


Giant blood cyst of tricuspid valve. Successful excision in an infant. A case of a giant blood-filled cyst of the tricuspid valve in described in a 4-month-old infant. The cyst caused obstruction of the right ventricular inflow and outflow tracts and a right-to-left shunt; it was successfully removed at open heart surgery. Pitfalls in differential diagnosis and the pathogenesis are discussed.


Cervical aortic arch and a new type of double aortic arch. Report of a case. A case of cervical aortic arch is reported. To the best of our knowledge, it is the first to be associated with a serious intracardiac anomaly. In addition, it is part of a new type of double aortic arch, caused by failure of reabsorption of both dorsal aortic roots and persistence of the fourth right and second (or third) left branchial arches.


Plasma proteins, packed-cell volume and requirements for alcuronium. The relationship between alcuronium requirements and the plasma concentration of albumin and total globulin was investigated in 50 adult patients undergoing elective upper abdominal surgery. No significant correlation was obtained with the plasma proteins (g/100 ml plasma). A significant positive correlation between alcuronium requirements and plasma albumin X (1-PCV), especially in anaemic patients, was found. This relationship tended to decrease with time.


Malignant hyperthermia in Belgian Landrace pigs rested or exercised before exposure to halothane. Thirteen of 31 Belgian Landrace pigs developed malignant hyperthermia (MH) after breathing halothane. A short period of exercise 1 h before the administration of the triggering agent increased the incidence of the syndrome to 100% in eight similar pigs. Clinical symptoms were more marked and developed more rapidly in the exercised pigs. All the reacting pigs became typically acidotic, developed rigor and died. Serum Na+, K+, Ca2+, c.p.k., l.d.h. and protein concentrations were increased to a variable extent during the reaction and there was an increase in p.c.v. also. No hyperglycaemia was detected in pigs which were rested before receiving halothane. Four of the eight exercised pigs became markedly hyperglycaemic and plasma noradrenaline increased to higher values. Phosphocreatine and ATP decreased to low values and lactate increased in the muscles of all pigs which reacted. At the time of death, muscle glycogen had decreased significantly in the rested, but not in the exercised, MH pigs.


Alveolar gas relationships during the use of semi-closed rebreathing anaesthetic systems. A black box and a geometrical approach have been used to deduce the relationships between alveolar oxygen and carbon dioxide concentrations when rebreathing occurs during the use of semi-closed anaesthetic rebreathing systems.


The hydrolysis of suxamethonium in human blood. In 30 patients, after induction of anaesthesia, the ulnar nerve on both arms was stimulated, and the resulting had contractions recorded or closely observed. Immediately after the circulation to one arm had been occluded with a tourniquet, suxamethonium 1 mg/kg was injected i.v. The occlusion was relieved after 1, 2 or 3 min. It was found that the duration of block on the occluded arm represented 68, 48 and 19% of the non-occluded arm after 1, 2 and 3 min occlusion respectively. Thus a considerable recirculation of the drug occurs, even 3 min after injection. It can be calculated that the disappearance of suxamethonium from blood is 7.7 mg/litre in the 1st min and 2.9 mg/litre in both the 2nd and the 3rd min. These disappearance rates include both hydrolysis and diffusion from plasma. Thus the rate for hydrolysis contrasts sharply with the published values for suxamethonium hydrolysis in vitro.


Plasma lignocaine concentrations following endotracheal spraying with an aerosol. Plasma concentrations of lignocaine were measured in three groups of anaesthetized patients following spraying of the trachea and larynx with a lignocaine 10% aerosol spray. Greater venous plasma concentrations occurred in patients who were paralysed with suxamethonium. A mean plasma concentration of 0.1 mug/ml of lignocaine resulted from each 10 mg of lignocaine used in spontaneously breathing patients, and 0.15 mug/ml in paralysed patients. In individual patients a concentration 50% in excess of the mean value may occur. The use of lignocaine 100 mg as a 10% aerosol spray can be considered safe.


Impairment of blood fibrinolytic activity during major surgical stress under combined extradural blockade and general anaesthesia. Blood fibrinolytic activity and plasma cortisol concentrations were investigated in 15 patients undergoing upper or lower abdominal surgery under high or low extradural blockade plus general anesthesia. It was found that the initial enhancement of fibrinolysis could be prevented but only in those patients showing impaired cortisol responses. This suggests a common nervous mechanism unrelated to the degree of sensory blockade.


The pulmonary capillary bed after lymphography. A small series of patients underwent radio-isotope lung scanning before and after the bipedal injection of oily contrast medium for abdominal lymphography to assess known malignant disease. After lymphography the lung scans showed an increase of between 12 and 100% in the radioactivity recorded over the anterior chest. No change was recorded over the posterior aspect of the chest. It is suggested that the increase is a result of blockage of posteriorly placed pulmonary capillaries and diversion of blood flow to the anterior segments.


A proinflammatory factor in lymphocytes. Its role in the development of acute, non-immunological inflammatory reactions. Drug-induced leucopenia renders rats hyporeactive to various inflammatory stimuli. Administration to leucopenic rats of suspensions of lymphocytes, sufficient to apparently correct the induced lymphocytopenia, led to a partial but marked reversal of the inhibited responses. Similar results were observed when lysates of lymphocytes or filtrates of the disintegrated cells were injected. Suspensions of polymorphonuclear granulocytes, on the contrary, were ineffective in producing a reversal of inhibited inflammatory reactions in leucopenic rats. The presence of a proinflammatory factor (LpIF) in lymphocytes, which might be involved in the modulation of acute inflammatory responses is suggested.


The effect of furosemide on the renal damage induced by toxic mushroom Cortinarius speciosissimus in the rat. The effect of furosemide on the renal damage induced by the toxic mushroom Cortinarius speciosissimus was studied in female rats. Furosemide (50 mg/kg s.c.) was given 15 min before the mushroom administration (250 mg dried mushroom/kg body weight p.o.). It was observed that furosemide clearly increased the tubular damage induced by C. speciosissimus. In contrast, furosemide had no effect on the inflammation induced by this toxic mushroom.


Effects of isoprenaline and pilocarpine on salivary glands as assessed by gravimetric and image analysis techniques. Pigs were given 6 daily injections of isoprenaline or pilocarpine. At necropsy the parotid, submaxillary and sublingual salivary glands were removed and weighed. Histological sections were prepared from several regions of each gland and the cross-sectional area of their acini was measured in treated and control animals by means of a Quantimet Image Analysis System. Both isoprenaline and pilocarpine produced a significant increase in the weight of the parotid and submaxillary glands, which was accompanied by an increase in acinar area. There was no change in the weight of the sublingual gland with either drug, but isoprenaline did induce an increase in the area of sublingual acini. A comparison is made of the changes in weight and structure of the salivary glands as shown by these methods.


Labelled polyvinylpyrrolidone as an in vivo indicator of reticuloendothelial activity. It is shown that after a single i.v. dose of [131I]-polyvinylpyrrolidone ([131I]-PVP) the total body and plasma radioactivities of rabbits decrease at distinctly different rates. The difference between these two rates is utilized to calculate the phagocytic rate of [131I]-PVP by reticuloendothelial cells.A number of experimental conditions are reported in which enhanced reticuloendothelial uptake of [131I]-PVP is readily demonstrable. They include the injection of small quantities of heterologous plasma, certain proteolytic fragments of the fibrinogen molecule, the clearance of antigen-antibody complexes, and the acute phase reaction (inflammatory response) as brought about by serum sickness, sterile abscess and vaccination. Based on these observations it is suggested that [131I]-PVP may provide a convenient technique for the long-term monitoring of the activity of reticuloendothelial cells, presumably mainly that of the histiocytes. The pronounced polydispersity of commercially available [131I]-PVP is a serious problem in this respect which can be largely overcome, but not completely abolished, by the screening techniques described herein. Post-mortem analyses of rabbit tissues showed most of the [131I]-PVP to be present in the skin (20%), followed by the liver (14%), bone marrow (10%), muscle (7%) and kidney (5%). Gel filtration studies with [131]-PVP in the presence and in the absence of plasma proteins failed to demonstrate any association between PVP and the proteins. [131I]-PVP kept at physiological pH and 37 degrees C lost less than 5% of its radioactivity over one month due to spontaneous deiodination.


Toxicity of various carrageenans in the mouse. Carrageenan toxicity was found to vary according to the biochemical nature and source of material injected. Ischaemic lesions of body extremities (acronecrosis) were produced by some but not all preparations. Histological examination of these sites and of both recent and old organ lesions (especially of liver and kidney) confirmed that the underlying pathology was disseminated intravascular coagulation. Evidence was also obtained that carrageenans are hepatotoxic.


Mouse hydrocephalus and encephalitis following intracerebral infection with Newcastle disease vaccine viruses. Hydrocephalus and encephalitis in 14-day-old mice was induced by an intracerebral inoculation of a high dose of live Newcastle disease vaccine viruses. The distended lateral ventricles occurred as a result of aqueduct stenosis superimposed on ciliary degeneration and destruction of ependymal cells. The brain virus titres were high on Days 3, 5 and 7 but never exceeded the titre of the original dose given. Positive serum haemagglutination-inhibition titres persisted throughout the experiment.


Metabolism of biliverdin. Biliary excretion of bile pigments after intravenous injection of biliverdin isomers. 14C-labelled biliverdins IX alpha, beta, gamma and delta have been prepared in vitro from haemoglobin obtained from duck erythrocytes incubated with 5-amino[4-14C]-laevulinic acid. When injected intravenously into rats with biliary fistulae, about 60% of the label was recovered in the bile in 24 h after the alpha isomer was given, while approximately 10% was recovered with injection of the beta isomer. The gamma and delta isomers gave intermediate values. In each experiment, most of the recovered isotope was found in association with conjugated bile pigment. Thus, the metabolic pathway for bile pigment excretion in the rat handles the IX alpha isomer preferentially but is not specific for it.


Structure of the three major sialyl-oligosaccharides excreted in the urine of five patients with three distinct inborn diseases: "I cell disease" and two new types of mucolipidosis. The urine of five patients with three distinct diseases ("I Cell disease" and two new types of mucolipidosis) contains sialic acid-rich oligosaccharides in a high amount: 50- to 500-fold the normal. The structure of the major components are as follows: alphaAcNeu(2 leads to 6)betaGal(1 leads to 4)betaGlcNac(1 leads to 2)alphaMan(1 leads to 3)betaMan(1 leads to 4)GlcNac,[alphaAcNeu(2 leads to 6)]betaGal(1 leads to 4)betaGlcNAc(1 leads to 2)alphaMan(1 leads to 3)[betaGal(1 leads to 4)betaGlcNac(1 leads to 2)alphaMan(1 leads to 6)]betaMan(1 leads to 4)GlcNAc and alphaAcNeu(2 leads to 6)betaGal(1 leads to 4)betaGlcNAc(1 leads to 2)alphaMan(1 leads to 3)[alphaAcNeu(2 leads to 6)betaGal(1 leads to 4)betaGlcNAc(1 leads to 2)alphaMan(1 leads to 6)]betaMan(1 leads to 4)GlcNAc. These results suggest that a deficit in alpha-neuraminidase is associated to these three different disorders and that an endo-beta-D-N-acetylglucosaminidase is able to release sialyoligosaccharides by splitting the sialylglycans of glycoproteins.


Effect of mucolytic agents on the rheological properties of tracheal mucus. Canine tracheal mucus was dissolved by a number of mucolytic agents, including disulfide bond reducing agents, hydrogen bond breaking agents, and chaotropic ions, and their effect on rheological properties was assessed. Sodium thiocyanate led to 85-100% dissolution with the maximum retention of elasticity. Thiocyanate exposure did not result in demonstrable alterations in the size or shape of the mucus glycoproteins. Sodium thiocyanate is therefore recommended as a suitable dispersing agent for physiochemical studies of glycoprotein secretions.


Copper deficiency and erythrocuprein (2Cu, 2Zn-superoxide dismutase). The activities of 2Cu,2Zn-superoxide dismutase, ferroxidase (ceruloplasmin), catalase and glutathione peroxidase were measured in the blood of rats during copper depletion. Two control groups of animals were used; one received the regular diet containing all essential components including copper and the other group was maintained on a diet, containing 1% the amount of copper in normal diet, copper being supplied as Cu(Leu)2 in the drinking water. Both groups showed no detectable differences, either in the copper content of blood or in the  measured four enzymic activities. Excessive copper (injected intraperitoneally) caused only an insignificant rise in the enzymic activities (0-10%) compared to either control. After starting copper depletion ferroxidase activity decreases to 15% on the 15th day, while the 2Cu,2Zn-superoxide dismutase activity decreases to 40% on the 45th day. Ferroxidase activity shows rapid but transient changes immediately after perturbation in plasma copper levels. By contrast, the 2Cu,2Zn-superoxide dismutase activity more closely parallels the overall copper deficiency. Dietary repletion with copper raises the 2Cu,2Zn-superoxide dismutase activity to 94% and the ferroxidase activity to 80% of the control values within 36 h. Apart from the copper-dependent anemia catalase activity was decreased. However, 15 days after the start of the copper depletion catalase activity rises again and reaches the control value on the 40th day and a 30% stimulation was even seen on the 58th day. Upon copper repletion catalase activity reaches 166% of the control within 14 days. No copper-dependent differences of glutathione peroxidase activity were seen regardless whatever copper level was present in the rats.


The stability of porphyrin and metalloporphyrin molecular complexes in solution. Stability constants for the molecular complexes formed by protoporphyrin IX dimethyl ester and its metal chelates with Co2+, Ni2+, Cu2+, Zn2+, Mg2+ and Fe3+ with nitroarene acceptors are determined in non-aqueous solvents. The stability of the complexes is found to be largely insensitive to variations in the central metal ions provided they are in the same oxidation state. Solvent-solute interactions are seen as being important in the formation of such complexes giving rise to large variations in their apparent stability. Some of the other factors that affect the stability of the molecular complexes are also evaluated. No evidence has been found for the appearance of any intermolecular charge-transfer absorption bands. Infrared studies, however, have provided some evidence for a substantial loss of electron density from the porphyrin ring upon interactions with the acceptors.


Transfer of heme from heme-albumin to hemopexin. Exchange of heme in vitro between two heme-binding serum proteins, albumin and hemopexin, was examined spectrophotometrically. Hemopexin, albumin and heme in molar ratios of 1 : 70 : 1 were incubated at 22 degrees C, pH 7.3. The heme was added as free heme, heme-hemopexin or methemalbumin. Due to the high affinity of hemopexin for heme, Kd near 10(-13) M, only negligible amounts of heme were transferred from hemopexin to albumin in 48 h. However, more than 80% of heme was transferred from methemalbumin to hemopexin within 24 h. Heme added to a 1 : 70 mixture of the apo-proteins is initially bound by albumin; but more than 90% is bound by hemopexin in 24 h. Addition of dithionite causes nearly all of the heme present, whether added as free heme or methemalbumin, to associate with hemopexin in 15 min. Albumin thus appears to have a much lower affinity for ferro- than for ferri-heme. Results obtained from similar experiments with human serum and human serum made hemopexin-free by immunoadsorption fully corroborate those obtained with mixtures of purified albumin and hemopexin. These observations suggest that the rate-limiting step in the heme transport function of hemopexin is the formation of the heme-hemopexin complex, rather than the uptake of the complex by the liver.


Influence of tryptophan on the level of hepatic microsomal cytochrome P-450 in well-fed normal, adrenalectomized, and phenobarbital-treated rats. In well-fed normal male rats, either force-feeding of tryptophan or a single injection of phenobarbital produced significant increments in hepatic microsomal cytochrome P-450 and the associated aniline hydroxylase activity. Administration of both tryptophan and phenobarbital together resulted in even greater stimulation than when the compounds were given alone. Adrenalectomy lowered instead the cytochrome P-450 concentration in comparison with that of normal rats, and administration of tryptophan and phenobarbital in this condition produced no significant increment in cytochrome P-450 concentration. In addition, phenobarbital administered either singly or in combination with tryptophan resulted in 80% mortality, which was reduced to zero by pretreatment with cortisol. While in cortisol-treated adrenalectomized rats administration of phenobarbital caused a 56% increment in cytochrome P-450 as compared to controls, tryptophan produced only a minor (9%) increase. In normal, as well as in adrenalectomized rats, tryptophan and phenobarbital administered either idividually or together increased microsomal protein concentration. In normal rats actinomycin-D treatment reduced both cytochrome P-450 and microsomal protein concentrations below that of the non-treated control levels. Further administration of either tryptophan or phenobarbital slightly increased the level of cytochrome P-450, and the two compounds together caused 40 and 21% increments of the same compared to actinomycin-treated and non-treated controls, respectively.


Multiple glucocorticoid binding components of intact AtT-20/D-1 mouse pituitary tumor cells. Glucocorticoids were found to bind to two components in the AtT-20/D-1 pituitary tumor cell. One component was characterized by slow dissociation of the bound steroid, stringent glucocorticoid specificity and high steroid binding affinity (Kd = 4.64 - 10(-9) M for triamcinolone acetonide). Thus, the characteristics of this component, termed the slowly dissociable component, resembled those of the soluble cytosol receptor. The other component exhbited lower binding affinity (Kd = 1.57 - 10(-7) M for triamcinolone acetonide), less stringent glucocorticoid specificity, and very rapid dissociation of bound, labelled glucocorticoid, Binding to this component, termed the rapidly dissociable component, represented 60% of total steroid binding to intact cells at 4 degrees C. Incubation of intact cells at 25 degrees C caused a progressive increase in steroid binding to the slowly dissociable component with no change in the absolute amount of ste roidal binding to the rapidly dissociable component. The high-binding affinity and preference for glucocorticoids shown by both components favor their function as biologically significant mediators of steroid action in this glucocorticoid responsive cell.


Heterogeneity of pituitary glucocorticoid binding evidence for a transcortin-like compound. Scatchard analysis, as well as sucrose gradient centrifugation demonstrate that at least two classes of specific glucocorticoid binding sites occur in the adenohypophysis. One, exhibited high affinity for both corticosterone and dexamethasone (receptor D), while the other bound only corticosterone. The latter binder revealed physico-chemical properties closely similar to those of plasma transcortin, but was shown not to be due to simple blood contamination of cytosol. This transcortin-like (TL) component, however, seemed to be carried over from plasma, since the concentration of pituitary binding sites strikingly paralleled physiological variations of plasma transcortin. Furthermore, experiments carried out with isolated pituitary cells suggested an intracellular location of that binder, although sticking to the cell membrane cannot be excluded. Finally, it was shown that the TL compound was unable, unlike the receptor D, to transfer glucocorticoids into the nuclei. One of its possible roles could be to modulate the interaction between steroids and compound D.


Establishment of the integrity of lysosomes in a glycoprotein-rich matrix. Distribution pattern of seven lysosomal enzymes in gastric mucosa. Gel-forming mucosal glycoproteins strongly interfere with standard methods of cell fractionation. Thus, acid hydrolase-bound particles imbedded in the gel, sediment on centrifugation, in the nuclear fraction of homogenates of canine antral mucosa. These particles can be cleared by direct solubilization of the gel; however, the viscosity of the solution obtained prevents sedimentation of some of the latent hydrolases, even at very high speeds. The use of a new step-wise scheme of centrifugation and dilution successfully isolates lysosomal particles containing acid hydrolases from mucin-rich mucosa. All of the enzymes investigated, including acid phosphatase, cathepsin D, alpha- and beta-galactosidase, beta-B-acetylhexosaminidases, but with the exception of alpha-fucosidase, were found to be particle bound, exhibiting high degrees of latency. However, active mucosal particles are polydisperase in size and density, sedimenting under different centrifugal forces.


Reduction by cycloheximide of lysosomal proteolytic enzyme activity and rate of protein degradation in organ-cultured hearts. Cycloheximide, an agent whose primary action is inhibition of protein synthesis, also causes a decrease in the rate of protein degradation in cultured fetal mouse hearts. This is associated with marked decreases in the activities of cathespin D and other lysosomal hydrolases. It is suggested that reduced lysosomal proteolytic capacity may contribute to cycloheximide-induced inhibition of protein degradation.


Isolation and characterization of 6-hydroxymethylpterin as the Crithidia growth-promoting factor from spinach chloroplasts. 2-Amino-4-hydroxypteridine compounds were isolated as the Crithidia factor from spinach chloroplasts by DEAE-Sephadex A-50, DEAE-cellulose, Sephadex G-25, and thin-layer chromatography. One of the compounds was characterized as 6-hydroxymethylpterin by gas-liquid chromatography-mass spectrometry and by comparison with authentic specimen.


Enthalphy changes associated with the denaturation of collagens of different imino acid content. The enthalpy changes associated with the denaturation of acid-soluble and insoluble collagens prepared from sheep, cod, halibut and pike skin were determined by differential scanning calorimetry. The enthalpy change associated with the soluble collagens decreased with decreasing imino acid content (from 1420 cal/mol for sheep to 736 cal/mol for cod) while the value for insoluble collagens was approximately constant at 1360 cal/mol. A possible explanation for these values in terms of the nautre of the bonds present in collagen is discussed.


The action of proteolytic enzymes on chloroplast thylakoid membranes. Envelope- and stroma-free thylakoid membranes of Vicia faba chloroplasts were incubated with trypsin or pronase for several hours. The indigestible residue was analysed by polyacrylamide gel electrophoresis. Trypsinization resulted in a complete digestion of all proteins with the exception of the pigment-protein complexes as well as a polypeptide not yet characterized. Yet, as compared with untreated material, Complex II was found to have higher electrophoretic mobility. Electron-microscopic studies illustrate that the indigestible residue still has a preserved membrane structure. Disintegration of the thylakoid membranes by sodium dodecyl sulfate followed by trypsinization also resulted in the two complexes while all the other proteins were found to be digested. However, after removal of the lipids the protein moieties of the complexes proved to be easily digestible. From these results it is concluded that pigment-protein interaction may be an important factor in maintaining a conformation rather resistant to perturbants and proteases. In contrast to trypsin, pronase completely digested the polypeptides of the thylakoid membranes including the protein moieties of the pigment-protein complexes leaving an amorphous lipid mass. The results support the assumption that the complexes are necessary to maintain the membrane structure.


Kinetics of permeation of nucleotide through oil/water interface in the interaction of nucleotide with octadecylamine and dodecyl guanidine. The transports of tritiated ATP, ADP and AMP from the aqueous to scintillator phase with and without octadecylamine (or dodecyl guanidine) have been studied by the layered scintillation method and a theory suitable for an explanation of the results has been presented. (1) Transport processes were all expressed by the first order kinetics. (2) For the simple partitioning of ATP, the reciprocal of the rate constant of the backward permeation was linear with respect to the square of the partition coefficient. (3) For the transport of nucleotide with chemical reaction, the reciprocal of the rate constant of the backward permeation was linear against the overall partition coefficient of nucleotide. (4) A theory was presented on the basis of a general diffusion equation by assuming the two-film model with potential energy near the interface. (5) The theory could explain the dependences of the permeation rates on the partition coefficients. (6) From the finding that the ratio of the apparent diffusion coefficient in aqueous to scintillator phase was much smaller than unity, the occurrence of an energy barrier at interface was suggested. For the simple partitioning of ATP, the energy barrier was not significant.


The binding of phenol red to rabbit renal cortex. The binding of phenol red to the microsomal fraction of rabbit kidney cortex was rapid, reversible and consisted of two independent populations of binding sites: a high affinity and low capacity class which had an association constant of 11.29 - 10(3) M-1 and a binding capacity of 2.41 mumol phenol red bound per g of protein, and a low affinity binding population with an association constant of 0.80 - 10(3) M-1 and a maximal binding capacity of 55.06 mumol per g of protein. Probenecid (0.32 mM) competitively inhibited phenol red binding to only the high affinity binding site, whereas 2,4-dinitrophenol (0.77 mM) competitively inhibited phenol red binding to both the high and the low affinity population of binding sites. The binding of phenol red was highly sensitive to the cationic composition of the medium. The affinity of phenol red to the high and the low affinity binding populations was lowered by decreasing the sodium and potassium concentrations to 19 and 6 mequiv./l, respectively; however, the maximal binding capacity was unchanged. Calcium appeared to have no effect on the phenol red binding to the microsomes. All of these considerations suggest that the high affinity phenol red binding to the microsomal fraction may represent the interaction of phenol red with the physiological receptor necessary for organic acid transport at the peritubular membrane. Phenol red binding to the low affinity binding population may indicate an intracellular binding population which contributes to the intracellular accumulation of weak organic acids.


Interactions between anaesthetics and lipid mixtures. Amines. The effect of a number of amine anaesthetics related to procaine on the temperature of lipid phase transitions has been studied using chlorophyll a as a fluorescence probe. The amines cause a reduction in the temperature of the phase transition of dipalmitoyl phosphatidylcholine and dipalmitoyl phosphatidylethanolamine and of mixtures of these lipids. The binding of charged amines causes a build up of positive charge on the membranes, limiting the binding. Incorporation of negative charge into the lipid bilayers causes a considerable increase in the binding of the charged amines, and the effect is reversed by addition of Ca2+. Anaesthesia is suggested to arise from an increase in the proportion of lipid in the liquid crystalline phase, resulting in a conformational change in the sodium channel. Effects of the tertiary amines on nerve conduction can be understood if the negatively charged lipid in the membrane is concentrated around the sodium channel: positively charged anaesthetics will have a greater effect when applied to the inside of a nerve because of the low Ca2+ concentration inside the nerve.


Carbohydrate composition of central nervous system synapses. Analysis of isolated synaptic junctional complexes and postsynaptic densities. The composition of specialized structures present at synapses within the central nervous system was elucidated by biochemical analysis of fractions enriched in synaptic junctional complexes and postsynaptic densities. The results indicate that the synaptic junctional complex is primarily protein together with some glycoproteins. The synaptic junctional complex proteins are similar in amino acid composition to synaptic membrane proteins; they are not expecially rich in basic residues, as previously suggested. The major carbohydrates present in the synaptic junctional complex and postsynaptic density glycoproteins are mannose, galactose, and glucosamine, with lesser amounts of fucose, N-acetylneuraminic acid, and galactosamine. Comparison with the synaptic membrane fraction indicates that galactose is more concentrated in the synaptic  junctional complex and mannose in the postsynaptic density. Glucose is dramatically enriched in both these fractions. Sucrose binding during isolation may partially account for the glucose enrichment.


X-ray diffraction study of myelin structure in immature and mutant mice. X-ray diffraction patterns were obtained from freshly dissected central and peripheral nerves of quaking, myelin synthesis deficiency (msd), and trembler mutants, as well as immature and adult normal mice. The patterns were compared with respect to strength of myelin diffraction, background scatter level, repeat period, and intensity and linewidth of Bragg reflections. The deficiency of myelin in optic nerves was found to be (in decreasing severity): quaking greater  immature greater trembler approximately normal adult; and in sciatic nerves: trembler greater immature greater quaking greater msd approximately normal adult. Repeat periods about 3 A less than that for normal adult sciatic myelin were detected in corresponding nerves from immature, quaking, and trembler mice. In some trembler sciatic nerves a second phase having a 190-200 A period and accounting for about 60% of the total ordered myelin was also evident. Comparison of electron density profiles of membrane units calculated from the repeat periods and diffracted intensities for sciatic myelins indicate structural differences at the molecular level. The main findings are: (1) quaking myelin shows a significant elevation of density in the external protein-water layer between membrane bilayers; (2) the membrane bilayer of immature myelin is approximately equal to 2 A thinner than that for normal adult; (3) the membrane bilayer of the more compact phase in trembler myelin is approximately equal to 5 A thinner than for normal; and (4) the difference in repeat periods for the two phases present in some of the trembler nerves can be accounted for predominantly by distinct membrane bilayer separations at the external boundary.


Energetics of permeation of thin lipid membranes by ions. The energy of an ion in a thin hydrocarbon membrane relative to its energy in a bulk aqueous phase is considered in terms of the electrostatic and surface components that may be expected to be involved. Except when diffusion activation energies are large compared to partition free energies, the latter will control permeation rate and the state of an ion having the lowest partition energy will be critical for its permeability. This minimum is found when an ion is surrounded with a thin layer of water. All ions of the same charge will tend to be at their lowest state in a sphere of water of the same size. It is concluded, therefore, that all ions of a given charge will have about the same permeability in lipid membranes.


Effects of unstirred layers or transport number discontinuities on the transient and steady-state current-voltage relationships of membranes. The effects of current-induced electrolyte accumulation and depletion on the electrical properties of a two-layered membrane system have been examined. The membrane consisted of a charged, ion permselective layer and an uncharged, non-selective layer. The model was designed to reveal the properties of membranes possessing long pores with ionic charges at one end or of ion-selective membranes bounded by highly unstirred aqueous layers. Electrolyte concentration profiles in the inert layer and their time-dependent changes were obtained from solutions of the diffusion equation under the condition of constant current. The profiles were then used to calculate the voltage developed across the membrane at various times after the current is switched on. The theoretical results are presented in the form of i-V curves with reduced coordinates that can be used to obtain time-current-voltage relationships for membranes of the type considered having any thickness of the non-selective layer and bathed in any concentration of any 1:1 electrolyte. Experimental results on a model composite membrane were in good agreement with calculations that assume that ion transport occurs only under the influence of electrical potential and concentration gradients, suggesting that in such systems, the combined effects of convection, osmosis, electro-osmosis, and concentration-dependence of diffusion coefficients, activity coefficients, and transference numbers are small. Voltage fluctuations in the form of periodic spikes were observed experimentally at the limiting current density (the current density at which the electrolyte concentration at one surface of the selective layer goes to 0). These phenomena were not seen when the current was in the direction leading to accumulation of electrolyte in the non-selective (unstirred) layer. Such composite membranes can exhibit S-shaped and N-shaped i-V curves under ramp-voltage and ramp-current clamps, respectively.


Phospholipid phase transitions. Effects of n-alcohols, n-monocarboxylic acids, phenylalkyl alcohols and quaternary ammonium compounds. The interactions of a series of alcohols, acids and quaternary ammonium salts with a phosphatidylcholine-water model biomembrane (dipalmitoyl phosphatidycholine) system have been studied using differential scanning calorimetry. In particular the effects of these molecules upon the lipid endothermic phase transitions were investigated over a range of concentrations. A variety of effects was observed. (a) Those molecules which shift or broaden the main lipid transition can also remove the pretransition endotherm. (b) n-Alcohols and n-monocarboxylic acids containing the same number of carbon atoms have very similar effects at molar concentrations up to 40%. Those molecules containing 12 or more carbon atoms raise the main lipid phase transition whilst those molecules containing 10 or less carbon atoms lower this transition temperature. (c) The phase diagram of stearoyl alcohol in the phosphatidylcholine-water system shows the formation of lipid-alcohol complexes. (d) Alkyl trimethyl ammonium bromides showed behaviour which differs considerably from n-alcohols and n-carboxylic acids of the same chain length. (e) Other alkyltrialkyl and tetraalkylammonium bromides show that a variety of effects on the lipid phase transition can be obtained. (f) With the homologous series of phenyl-alkyl alcohols from benzyl alcohol to 4-phenyl butanol increasing the number of methylenes between the terminal OH and the benzene ring leads to greater interaction between solute and bilayer. The range of different effects obtained with the compounds studied offers a means for introducing various degrees and types of perturbation into membrane systems.


Studies on membrane fusion. I. Interactions of pure phospholipid membranes and the effect of myristic acid, lysolecithin, proteins and dimethylsulfoxide. The interaction and mixing of membrane components in sonicated unilamellar vesicles and also non-sonicated multilamellar vesicles prepared from highly purified phospholipids suspended in NaCl solutions has been examined. Electron microscopy and differential scanning calorimetry were used to characterize the extent and kinetics of mixing of membrane components between different vesicle populations. No appreciable fusion was detected between populations of non-sonicated phospholipid vesicles incubated in aqueous salt (NaCl) solutions. Mixing of vesicle membrane components via diffusion of phospholipid molecules between vesicles was observed in populations of negatively charged phosphatidylglycerol vesicles but similar exchange diffusion was not detected in populations of neutral phosphatidylcholine vesicles. Incubation of sonicated vesicle populations at temperatures close to or above the phospholipid transition temperature resulted in an increase in vesicle size and mixing of vesicle membrane components as determined by a gradual change in the thermotropic properties of the mixed vesicle population. The interaction of purified phospholipid vesicles was also examined in the presence of myristic acid and lysolecithin. Our results indicate that while these agents enhance mixing of vesicle membrane components, in most cases mixing probably proceeds via diffusion of phospholipid molecules rather than by fusion of entire vesicles. Increased mixing of vesicle membrane components was also produced when vesicles were prepared containing a purified hydrophobic protein (myelin proteolipid apoprotein) or were incubated in the presence of dimethylsulfoxide. In these two systems, however, the evidence suggests that mixing of membrane components results from the fusion of entire vesicles.


The transport of calcium and sodium ions across bilayers induced by neutral synthetic ligands used in specific electrodes. Neutral synthetic ligands of calcium and sodium which enter into the liquid membrane composition of the selective electrodes of these ions have been incorporated within the bilayer. The membrane conductance measured shows that each of these two ligands behave as ion carriers for calcium or sodium ions. The selectivities with respect to the other alcaline or alcaline earth ions are similar to those observed by potentiometric measurements with thick liquid membrane selective electrodes.


Effects of cholera toxin on cellular and paracellular sodium fluxes in rabbit ileum. The diarrhea observed in patients with cholera is known to be related to secretion of water and electrolytes into the intestinal lumen. However, the exact mechanisms involved in these secretory processes have remained unclear. Although it is clear that purified toxin acts on epithelial cell metabolism, its activity on Na+ transport across intestinal mucosa is equivocal: reported either to prevent net Na+ absorption or to cause net secretion of Na+ from serosa to mucosa. Since total transmural Na+ fluxes across "leaky" epithelia involve very significant movement via a paracellular shunt pathway, we studied the effects of cholera toxin on the cellular and paracellular pathways of Na+ movement. Unidirectional Na+ fluxes were examined as functions of applied potential in control tissues and in tissues from the same animal treated with purified cholera toxin. Treatment of rabbit ileum in vitro with toxin simulated the cellular component of serosa-to-mucosa Na+ flux (from 2.41 +/- 0.49 muequiv./h per cm2 under control conditions to 4.71 +/- 0.43 muequiv./h per cm2 after treatment with toxin, P less than 0.01). The effect of cholera toxin on Na+ movement through the cells from mucosa to serosa appeared to be insignificant. Finally, a marked decrease in the Na+ permeability (P less than 0.01) and no detectable significant changes in transference number for Na+ of the paracellular shunt pathway were observed following treatment with cholera toxin. These results provide direct evidence for the hypothesis that purified cholera toxin stimulates active sodium secretion but has minimal effect on sodium absorption.


On the susceptibility of human platelets to aggregation by concanavalin A and the effect of this lectin on their response to ADP. Concanavalin A aggregated gel-filtered platelets in 0.9% NaCl solution signifying cross-bridging by the lectin. Aggregation of these platelets by concanavalin A was temperature dependent; it did not occur at 0-4 degrees C unless the platelets were previously trypsinized. The level of aggregation of trypsinized platelets by concanavalin A at 0-4 degrees C was similar to that of untreated platelets at 37 degrees C. It is suggested that trypsin facilitates platelet aggregation by concanavalin A at 0-4 degrees C by causing a configurational change in membrane glycoproteins which orientates concanavalin A receptor sites into positions that favour lectin cross-bridging. Concanavalin A failed to aggregate platelets in plasma. Radioisotope studies showed that the amount of [3H]concanavalin A which combined with platelets in plasma was extremely low compared with gel-filtered platelets in saline. The aggregation of Ehrlich ascites cells by concanavalin A was considerably reduced when platelet-free plasma was added to the medium suggesting that it was due to the presence of concanavalin A-reactive components in the plasma. Concanavalin A inhibited the ADP-induced aggregation of platelets suspended in plasma or in a salts solution supplemented with calcium and fibrinogen, although the inhibitory effect was more conspicuous in the latter case. The results suggests that concanavalin A produces its inhibitory effect on ADP-induced platelet aggregation by interacting with membrane glycoproteins, and this further suggests their involvement in aggregation.


The thermodynamic degree of coupling between metabolism and sodium transport in frog skin. The tightness of coupling between two processes is advantageously evaluated by the thermodynamic degree of coupling q, varying in absolute value from zero for uncoupled processes to unity for processes which are related stoichiometrically. Two methods for the determination of q in the active pathway in frog skin have been developed, employing amiloride to abolish active sodium transport. The values of q in 6 frog skins varied, but were always less than unity (mean 0.79 +/- 0.06 S.E. according to one method, 0.78 +/- 0.06 S.E. according to the other). This indicates that metabolism and sodium transport are incompletely coupled in this tissue even when passive transepithelial leakage pathways are taken into account.


Cell membrane fluidity: molecular modeling of particle aggregations seen in electron microscopy. The first simulation of the fluid mosaic model is reported. Intra-membrane particles, initially "placed" randomly in a membrane with fluid properties, are allowed to diffuse in the plane of the membrane and to interact with one another, in a model using molecular parameters. The resulting particle aggregates are very similar to those observed in freeze-fracture electron microscopy.


A.E. Bennett Award Paper. A kinetic analysis of 5-hydroxyindoleacetic acid excretion from rat brain and csf. Studies of neurotransmitter kinetics based on intraventricular injections of radio-labeled metabolites have been limited by several problems, including the inability of most investigators to recover more than 45% of the infected isotope from brain homogenates within several minutes after the injection...


Bipolar course in schizo-affective illness. The authors undertook a chart review to examine the use of the diagnosis of schizophrenia, schizo-affective type, in clinical practice. Of 27 patients given this diagnosis over a 3-year period, 13 were found to have evidence of bipolar course in their illnesses. For both the bipolar and unipolar groups, the most striking finding in first-degree relatives was the prominence of affective conditions. The bipolar group had a statistically significant earlier age for first psychiatric treatment and previous number of hospitalizations. Symptoms noted on admission were mostly affective, and the schizophrenic symptoms reported were noted by authors to be considered overinclusive or unreliable by many clinicians. Both groups received treatment with antipsychotic and antidepressant medication. Six of 13 bipolar patients and no unipolar patients were treated with lithium carbonate. Five bipolar patients met research criteria of Feighner et al. (1972) for primary affective illness and another met the criteria for schizophreniform illness. One unipolar patient met criteria for probable primary affective illness and another met the criteria for probable schizophreniform illness. The authors concluded that the diagnosis of schizo-affective illness, as used in day to day clinical practice, does not identify a group of schizophrenic patients nor a homogeneous patient group, and when both affective and schizophrenic features appear in a patient with a bipolar illness, the diagnosis of manic-depressive illness, not schizophrenia, should be given first consideration.


Visibility of the nailfold plexus and heredity. The parents of 37 normal healthy teenagers selected on the basis of nailfold capillary patterns (16 low-plexus children and 21 high-plexus children) were examined by capillary microscopy and rated "blindly" for plexus visualization scores (PVS). The hypothesis that high PVS is a hereditary trait transmitted as a monogenic autosomal dominant is supported by these data. The additional hypothesis that the high PVS may be associated with a higher social class is also confirmed in this sample. The transmission of high PVS in schizophrenics, where it is frequently observed in association with more serious features of this disease, appears to follow the same mode of inheritance as in normal subjects. The importance of detecting a biological hereditary characteristic which appears to have a modifying effect on the schizophrenic process is emphasized.


14C-homovanillic acid in the cerebrospinal fluid of parkinsonian patients after intravenous 14C-L-dopa. Six patients with Parkinson's disease and five controls were premedicated with probenecid and the peripheral decarboxylase inhibitor alpha-methyldopathydrazine (Carbidopa) before intravenous administration of 50 muc of 14C-L-dopa in tracer quantity. Seven-and-one-half hours later lumbar CSF was obtained. 14C-homovanillic acid (HVA), a major metabolite of brain dopamine, was isolated by thin-layer chromatography and measured. The statistically significant positive correlation between endogenous HVA and 14C-HVA in the entire patient group and the slightly lower values of endogenous HVA and 14C-HVA in the CSF of the parkinsonians support the assumption that the concentration of HVA in the CSF after probenecid treatment reflects brain dopamine turnover. Measurement of labeled HVA here seems to have little advantage over measurement of endogenous HVA alone.


Effects of changes in carbon dioxide tension in abnormally prolonged contingent negative variation (CNV). The effects of changes in pCO2 in the development and resolution of normal (return to base line within 2 sec after S2) and abnormal (return to base line in more than 2 sec after S2) CNVs were studied. Decreased pCO2 produced by hyperventilation induced a significant reduction in the duration of the post-imperative negative variation (PINV) without significantly affecting the amplitude of the CNV. Increased pCO2, by inhalation of 5% CO2, on the contrary, produced prolongation of the PINV in two subjects. Control subjects with normal CNVs did not show significant changes in the amplitude or in the duration of the CNV wave when pCO2 was altered. The results are discussed in terms of the possible independence and higher sensitivity to metabolic changes of the PINV brain generators compared with the generators of the CNV wave per se.


Diagnostic implications of the B-mitten EEG pattern: relationship to primary and secondary affective dysregulation. Previous studies indicate that the B-Mitten EEG pattern is specific to psychiatric dysfunction and rarely encountered among normal control subjects. Our previous investigations have demonstrated a significant relationship between mittens and reactive as opposed to process schizophrenic disease. Re-analyses of mitten incidence in schizophrenics and nonschizophrenics suggests needed modification of our earlier B-Mitten-schizophrenia formulation. Present considerations indicate that the B-Mitten-reactive schizophrenia association is not primary but rather that the differential process-reactive schizophrenia mitten incidence may be a secondary epiphenomenon reflecting a more fundamental underlying process. Analyses are presented suggesting that the mitten dysrhythmia relates positively to dysphoric affective dysregulation in a manner which cuts across broad diagnostic boundaries. That the mitten pattern possibly suggests as yet unclarified subcortical dysfunction associated with symptoms of affective disturbance is a tentative hypothesis offered for consideration.


Electrophoretic pattern of red blood cell catechol-o-methyltransferase in schizophrenia and manic-depressive illness. Human red blood cell (RBC) catechol-O-methyltransferase (COMT) was analyzed by polyacrylamide gel electrophoresis (PAGE). One major enzyme band (B) is observed after electrophoresis. In addition, a minor band (A) of COMT activity comprising no more than 25% of the total activity, is also detectable. The rate of migration during electrophoresis of both bands of RBC COMT was the same in manic depressive, schizophrenic, and normal individuals. These results did not reveal genetic variations in the COMT molecule among these three groups. Furthermore, when total RBC COMT was measured there were no statistically significant differences between schizophrenic, manic-depressive, and control individuals.


Comparison of etidocaine and lidocaine for obstetrical analgesia. Etidocaine, a relatively new local anaesthetic agent, is compared with lidocaine for use in epidural anaesthesia for vaginal delivery. The parameters compared were quality of anaesthesia, dose of drug, onset and regression of sensory blockade, complete degree of motor blockade, effect on maternal blood pressure, and foetal Apgar scores at one and five minutes. The major differences between the drugs are the lower percentage of satisfactory anaesthesia with etidocaine, the much greater variability in the time of onset of complete sensory block with etidocaine as compared to lidocaine, the more profound motor block with etidocaine, and the slightly longer duration of action of this drug. It is concluded that etidocaine has no real place in epidural anaesthesia for obstetrics.


Components of respiratory depression after narcotic premedication in adolescents. The effects of narcotics on ventilatory control were assessed in 13 adolescents and young adults. Both a narcotic and narcotic-phenothiazine significantly depressed the CO2 response curve. Using an occlusion pressure technique (Pm100) to evaluate those neuromuscular processes that generate forces acting on the ventilatory pump, it was found that narcotic agents reduced neuromuscular drive. In most subjects, narcotics had an additional action that contributed to the overall ventilatory depression. Using carbon dioxide to vary neuromuscular drive before and after drug administration at constant levels of neuromuscular drive the drugs reduced tidal-volume responsiveness of the pump. We conclude that narcotics impair ventilation through a combination of two effects; first, reduced neuromuscular drive, most probably due to central depression, and second, increased impedance of the ventilatory pump, most probably due to a decrease in chest-wall compliance.


Toxicity following methoxyflurane anaesthesia. IV. The role of obesity and the effect of low dose anaesthesia on fluoride metabolism and renal function. Seven obese and five normal weight patients were studied before, during and after one hour of methoxyflurane-nitrous oxide anaesthesia during peripheral surgical operations and compared with eight patients of normal weight anaesthetized with nitrous oxide-meperidine and d-tubocurare. Estimates were made of renal function, including serum and urinary electrolytes, osmolarity, uric acid, urea and creatinine. Renal clearances for the latter three substances were also calculated. Serum and urinary inorganic and organic fluoride concentrations were measured, as were renal clearances. This low dose methoxyflurane anaesthesia resulted only in a decrease in uric acid clearance among all the measures, when compared to the meperidine-nitrous oxide controls. The clearance of uric acid remained depressed for longer in the obese patients, but otherwise they did not differ from the normal weight patients. It is possible but not proven that depressed uric acid clearance may be related to the organic fluoride metabolite and an early indicator of methoxyflurane renal toxicity. The previously documented biotransformation of methoxyflurane was seen in this study. A double peak in serum inorganic fluoride was shown in all patients but one. Rather large differences in peak levels of serum inorganic fluoride occurred. The only significant difference between the obese and normal weight patients as far as fluoride metabolism was concerned was a greater variability in the serum inorganic fluoride levels in the obese patients. It would appear that the obese patient metabolizes methoxyflurane in a quantitatively if not qualitatively different fashion than the normal weight patient, perhaps because of fatty infiltration of the liver. Caution is advised in the use of methoxyflurane for more than 90 minutes of low concentration administration in view of the unpredictability of the biotransformation.


Effect of anaesthesia and surgery on blood sugar and carbohydrate tolerance in African children. The effect of pre-operative starvation, anaesthesia and surgery on blood sugar levels and the handling of carbohydrate load during operation were studied in 28 Nigerian children between 2 months and 15 years of age. (1) Age and body weight were important factors influencing the relationship between duration of pre-operative fast and the pre-induction blood sugar level in children. Hypoglycaemic values occurred in 7 per cent of the subjects studied although none was clinically hypoglycaemic. (2) Halothane anaesthesia alone did not affect blood sugar levels but relaxant anaesthesia in this study caused significant rise of blood sugar. (3) There was a marked hyperglycaemic response to surgery and handling of glucose load during operation was significantly poorer than before operation.


[Etomidate: cardiovascular effects of a new intravenous anesthetic agent]. The cardiovascular effects of intravenous Etomidate (0.2 mg/kg) were studied in 14 surgical patients with ages varying from 49 to 90. The anaesthetic induction occurs within 10 seconds and the mean duration of anaesthesia is from 6 to 8 minutes. I.V. Etomidate causes a slight lowering of the mean arterial pressure (8.5 per cent), a negligible increase of the heart rate (2.8 per cent) and an insignificant lowering of the mean pulmonary artery pressure (7 per cent). Cardiac output and stroke volume are respectively lowered by 7.6 per cent and 10 per cent. The peripheral vascular resistance is discreetly reduced by 3.8 per cent. From these results, we believe that Etomidate seems to induce sleep with the lightest hemodynamic disturbances in comparison with other agents commonly used. However, some undesirable side effects appeared in some of our patients which can become somewhat annoying, for example myoclonic movements and pain at the point of injection.


Ten cases of malignant hyperthermia in Norway. Data are presented on ten cases of anaesthesia-induced malignant hyperthermia in Norway. Seven of the patients died, three recovered. The fatal cases were all boys in the age group 11-20 years. This age and sex distribution suggests that puberty with the increase in androgens is a precipitating factor in malignant hyperthermia. One of the victims who survived was a 4 1/2-year-old pseudohermaphrodite girl with the adrenogenital syndrome. The coincidence of malignant hyperthermia in a patient with such a rare syndrome points to the excessive formation of androgens in patients with this syndrome as a predisposing factor. The indications for surgery were traumatic injuries in five cases, congenital abnormalities in three and appendicitis in two cases. These conditions in themselves may cause an increased sensitivity to suxamethonium. One patient received only hexobarbitone, halothane and suxamethonium. After the last drug jaw rigidity and temperature rise to 41.3 degrees C prompted the anaesthetist to end the anaesthetic. The fact that the patient survived proves that suxamethonium induced jaw rigidity is valuable as a warning. The absence of cardiovascular depression after procaine 3.5 g in one patient is ascribed to the correction of acidosis at the time of infusion of this drug. It is suggested that procaine should be withheld until other measures such as cooling, correction of acidosis and steroid therapy have been tried.


The inflation-catheter technique of ventilation during laryngoscopy. Over the past three years the inflation-catheter technique (ICT) of controlled ventilation during general anaesthesia has proved a safe and useful method for use with laryngoscopy and other trans-oral endoscopies (most notably flexible fiberoptic bronchoscopy). The ICT allows full control of the patient's airway for the anaesthetist and exceptionally good exposure for the surgeon. In this technique ventilation of the patient is accomplished by intermittent high flow (10 ml/kg/sec) inflation of the lungs through a small (2 to 6 mm diameter) plastic orotracheal catheter. The inflation-catheter is stiffened by fixation of an intralumenal wire. Exhalation is passive around the catheter through the open airway. The Inflation Catheter Technique is readily adaptable for patients ranging from large adults to small babies by variation of catheter sizes. It can be used safely with nitrous oxide-oxygen as the ventilating gas(i.e., Bird N2O/O2 Blender) since air entrainment is not a feature. It is also readily adaptable for use with mechanical ventilation (i.e., Wolf Injectomat).


A latex bag tonometer for determining the anaesthetic blood/gas partition coefficient. Many techniques for measurement of the tissue/gas partition coefficient are valid. Each has specific advantages and shortcomings. All the methods require suitable analytic equipment and means for controlling temperature. The disadvantages of a particular technique, the ability to cope with them and the availability of the necessary apparatus are the determining factors in choosing the best method. The latex bag tonometer has a precision and ease which compares favourably with previously described techniques for measuring anaesthetic blood/gas partition coefficients. The technique was successfully used to show the effect of in vivo haemodilution on the halothane blood/gas partition coefficient.


The selective extraction of histones from rye chromatin. The selective extraction of histones from rye chromatin was studied using three different methods. Extractions with NaCl-phosphate buffers at pH5.5 gave results similar to those already obtained with other types of chromatin. Histone H1 was selectively extracted with 0.6 M NaCl-0.001 M PO4, while the selectivity of dissociation of the other fractions was reduced at higher NaCl concentrations. The use of phosphate-urea buffers at pH5.5 also revealed that the histones were dissociated at the same concentrations as were calf thymus histones. Histone H1 was extracted with 0.5 M PO4-1 M urea; H1, H2A, and H2B were extracted with 0.8 M PO4-2 M urea; and all histones were removed with 0.8 M PO4-5.3 M urea. It was, however, observed that the dissociated rye histones H2A and H2B were unstable in these buffers. This instability was maximum in the presence of 3 M urea, where both histones were absent from the extracted proteins and the residual nucleoproteins. Finally, a solution of 30% ethanol-0.35 M NaCl-6 M urea produced a rye nucleoprotein fraction containing only histone H1.


GM1-ganglioside and lactosylceramide beta-galactosidase from rabbit brain: inhibitor and substrate competition studies. A rabbit brain beta-galactosidase catalyzes the hydrolysis of synthetic substrates and the natural substrates Gm1-ganglioside, lactosylceramide, and asialo-Gm1-ganglioside. gamma-D-Galactonolactone competitively inhibited hydrolysis of Gm1-ganglioside, lactosylceramide, and MU-galactoside with Ki values of 0.26 mM, 0.13 mM, and 0.77 mM, respectively. From activity plots comparing the degree of inhibition to the inhibitor concentration, a single binding site for each substrate was found. NP-Galactoside inhibited the hydrolysis of Gm1-ganglioside and lactosylceramide, where as Gm1-ganglioside inhibited lactosylceramide hydrolysis. At low substrate concentrations (less than 1 mM), Gm1-ganglioside was hydrolyzed effectively in the presence of NP-galactoside, but at higher concentrations hydrolysis of the latter was preferred. Chloromercuriphenylsulfonic acid and iodoacetate were effective inhibitors of the enzyme, but N-ethylmaleimide was not. The degree of inhibition with chloromercuriphenylsulfonic acid was different for each substrate. At 0.5 mugM chloromercuriphenylsulfonic acid, all activity towards NP-galactoside, 75% towards lactosylceramide, and 25% of the Gm1-ganglioside activity was lost. Two possible models are presented to explain these results. The data favour the presence of multiple active sites in the enzyme.


Enzymatic synthesis of C40 carotenes by cell-free preparation from Halobacterium cutirubrum. [14C]Mevalonate or (14C)isopentenyl pyrophosphate was found to be converted to transphytoene, trans-phytofluene, lycopene, and beta-carotene by a cell-free 270 000 X g supernatant fraction prepared from Halobacterium cutirubrum cells that were broken by manual grinding with glass beads. Incubations were done under N2 in the dark at 37 degrees C in 4 M NaCl in presence of FAD, NADP, and MgCl2; ATP was also added when mevalonate was the substrate. This system was also capable of converting trans-(14C)phytoene to beta-carotene via the intermediates trans-phytofluene, zeta-carotene, neurosporene, lycopene, and gamma-carotene. Each of these labelled intermediates on incubation separately with the same enzyme system was shown to be converted to the intermediates farther down the pathway. The results of this study show that the biosynthetic pathway for the formation of C40 carotenes in H. cutirubrum proceeds as follows: isopentenyl pyrophosphate leads to trans-phytoene leads to trans-phytofluene leads to zeta-carotene leads to neurosporene leads to lycopene leads to gamma-carotene leads to beta-carotene. This pathway differs from that in higher plants in that the cis isomers of phytoene and phytofluene are not on the main pathway of carotene biosynthesis, as they are in higher plants. Furthermore, trans-phytoene, which has not been reported to have any role in higher plants, appears to be the main intermediate in carotene biosynthesis in H. cutirubrum.


Effect of nicotine on biosynthesis of C50 carotenoids in Halobacterium cutirubrum. The major carotenoids in pigmented extreme halophiles are the hydroxylated C50 red pigments bacterioruberin (major, 84%) and monoanhydrobacterioruberin (minor, 14%). When cells of Halobacterium cutirubrum were grown in the presence of nicotine they accumulated lycopene and bisanhydrobacterioruberin; maximal accumulation (80% lycopene, 20% bisanhydrobacterioruberin) occurred at 3mM nicotine. Removal of nicotine resulted in the reformation of monoanhydrobacterioruberin and bacterioruberin at the expense of lycopene and bisanhydrobacterioruberin. Thus nicotine does not inhibit the addition of the two extra isoprene units plus two hydroxyl groups (at C1 and C1') to a C40 carotenoid skeleton. It does, however, inhibit the hydration steps at C3'' and C3''' that convert bisanhydrobacterioruberin to monoanhydrobacterioruberin and finally bacterioruberin. These results support the view that the C50 carotenoids can be formed from a C40 carotene, lycopene.


Reduction of pulsatile hydraulic power in the pulmonary circultation caused by moderate vasoconstriction. Vascular input impedance and associated hydraulic power was measured in rabbit isolated lungs. The study was focused on changes in impedance and in pulsatile hydraulic power during relaxation and contraction of vascular smooth muscle. Pulsatile power was found to be at a minimum when smooth muscle tone was such that the pulmonary arterial pressure was in the physiological range, and increased both when the vessels were relaxed and further constricted. Input impedance was found to be determined mainly by the large, proximal ('extra-alveolar') arteries.


Myocardial contractile reserve and indices of contractility. A close examiniation of several indices of contractility derived from the ventricular pressure was made using an isolated canine heart preparation. The repsonses to single intracoronary injections of calcium chloride as well as to increasing doses of this agent were tested. From the latter, calcium dose-transient-response curves intended to reveal the extent of myocardial/contractile reserve were constructed. Indices included various extrapolations to maximal velocity of shortening at zero load, maximal value of the quotient of the first derivative and ventricular pressure, time-tension index, energy averaged power density, and power averaged rate of generation of power density. Indices were compared over the same cardiac cycles before and after administration of calcium. Most indices showed increments from 10 to 20%, except power density functions which had increments of 40 and 70%, respectively. Calcium dose-response curves were linear for most indices, but, again, the power density functions showed the steepest slopes. After severe coronary occlusion, the curves for most indices flattened and lost linearity and, presumably, this was due to loss of myocardial contractile reserve. For milder occlusions, only the power density functions showed significant flattening. The conceptual framework of a contractile myocardial reserve appears supported by these results.


Influence of procainamide on sodium and potassium exchange and permeabilities in cultured human cells. The effect of procainamide on membrane cation exchange was investigated using monolayer cultures of Girardi heart cells. The initial effect of procainamide (10(-6) to 10(-3) mol/litre) was to produce a prompt reduction of the passive Na influx, dose-dependent along a sigmoid log dose-response curve. This effect was complete within 3 min and thereafter showed no further time-dependent increase. Mean passive Na influx (pmol-cm-2/s) decreased from 19.1 to 17.7 (P less than 0.05) and 10.4 (P less than 0.001) in 10(-5) and 10(-3) mol/litre procainamide, respectively. No effect on active Na extrusion was noted before 3 min following exposure to the drug, after which time it progressively declined reaching a minimum value for each concentration by 6 min and remaining at this level throughout a further 60 min exposure. For each concetnration this minimum value was similar to the Na influx measured under identical conditions. Na-coupled active K influx showed a parallel pattern of inhibition. K efflux was not decreased until approximately 20 min following exposure to the drug, but once present the reduction was similar in magnitude to that in the correspondingly measured K influx. Kinetic flux analysis revealed a decrease in both PNa and PK but indicated a greater effect on PNa. The results suggest that all of the above effects could be explained on the basis of one direct action of the drug, namely, the prompt initial decrease in PNa and Na influx. All other effects noted, both active and passive, could then be secondary to this phenomenon.


Progressive reduction in norepinephrine overflow during cardiac sympathetic nerve stimulation in the anaesthetized dog. The left ansa subclavia was stimulated at supramaximal intensity at a frequency of Hz in open-chest, anaesthetized dogs for periods of about 10 min. Arterial blood pressure, myocardial contractile force, and coronary sinus blood flow reached peak values after 1 or 2 min of stimulation, and then declined gradually throughout the remainder of stimulation. In hearts that averaged 159 +/- 10 (SEM) g, the norepinephrine (NE) overflow rose to a peak value of 470 +/- 128 ng/min at the end of 1 min of sympathetic stimulation in the control animals. By the 10th min of stimulation, the NE overflow had diminished to 97 +/- 24 ng/min. In animals pretreated with cocaine (5 mg/kg) or with phenoxybenzamine (5 mg/kg), the peak NE overflow rates were 288 +/- 62 and 980 +/- 148 mg/min, respectively, and the rate of NE overflow declined to levels of 104 +/- 39 and 128 +/- 30 ng/min, respectively by the 10th min of stimulation. Since there was a profound, progressive diminution in the rate of NE overflow during sustained cardiac sympathetic stimulation regardless of whether or not NE reuptake was suppressed, it is concluded that the reduction in NE overflow reflects a curtailment of neuronal release of NE rather than an acceleration of reuptake.


On-line computation of cardiac output with the thermodilution method, using a digital minicomputer. In the present study, several techniques for calculating cardiac output were evaluated in order to find an accurate technique suitable for on-line digital computation. The thermodilution method was evaluated by different calculation techniques and by comparing these techniques with electromagnetic flowmeter values. Comparison of thermal dilution curves, manually calculated by the log-normal and the exponential assumption, showed a correlation coefficient of 0.978 between the two methods, the former values being 11% higher. The exponential method correlated very well with the technique, using a correction factor for injection errors (r = 0.999). Since the exponential technique correlated well with the electromagnetic values (r = 0.84) and since it was less complicated than the technique, using correction factors, this technique was chosen for automation. Comparison of exponential calculated thermodilution values with a digital computer and manually, showed a correlation coefficient of 0.991. Therefore, it was concluded that on-line computation of thermodilution curves improves the applicability of the termodilution techniques as a means of measuring cardiac output.


Reflection in the systemic arterial system: effects of aortic and carotid occlusion. Experiments were performed in seven closed-chest anaesthetized male dogs to determine the role of pulse wave reflection in the pattern of flow and pressure in the ascending aorta. Ten days after implantation of an electromagnetic flow transducer around the ascending aorta a balloon catheter was placed in the descending aorta via the femoral arteries. At the same time a tip manometer was introduced into the ascending aorta. Aortic occlusions at three different sites caused pressure pulses with secondary systolic rises and flow pulses with biphasic deceleration. Secondary rises occurred 45 +/- 9.0 ms after the initial pressure rise for high aortic occlusion; this time was 75 +/- 8.5 ms for occlusion at the level of the diaphragm and 114 +/- 16.5 ms for occlusion near the level of the renal arteries. These times approximate the times in which the pulse travels from the tip manometer to the inflated balloons and back. Forward and reflected pressure and flow waves were calculated from reflection coefficients. Aortic occlusion caused larger reflected waves and the recorded wave forms were caused by the summation of forward and backward waves, the latter contributing the secondary pressure rise and the increased flow deceleration. Occlusion of both carotid arteries showed no specific reflection site but reflected waves were larger. This increased reflection can probably be explained as the result of greater total reflection from distributed sites under increased peripheral resistance.


Effect of ischaemia on overdrive suppression in isolated, blood-perfused atrial preparations of dogs. The magnitude of overdrive suppression of the SA nodal pacemaker activity was markedly influenced by its ischaemia which was produced by occlusion of the perfusion tubing close to the sinus node artery in the in-situ and isolated canine sino-atrial node preparations. In all isolated preparations, postdrive suppression was significantly augmented after occlusion of the perfusion system for at least 120 seconds. It is concluded that total ischaemia depresses sino-atrial node function in the atropinized canine heart and that postdrive suppression is a good index of such impaired function.


Controls of ventricular contractility assessed by pressure-volume ration, Emax. The present study on canine left ventricles showed that Emax which we previously proposed as a good index of both ventricular contractility and its pumping capability, decreased from 2.06 to 1.38 kPa/ml (15.5 to 10.4 mmHg/ml) via the sino-aortic baroreceptor reflex. Cerebral ischaemic response increased Emax to 3.83 kPa/ml (28.8 mmHg/ml). Emax decreased to 1.17 kPa/ml (8.8 mmHg/ml) after cardiac denervation.


Protective effect of hyperbaric oxygen for the temporary ischaemic myocardium. Macroscopic and histological data. A protective effect of hyperbaric oxygen immediately after reopening of occluded coronary blood flow for the temporary ischaemic myocardial muscle was studied. Thirty dogs were used in this study, and 20 dogs were sacrificed after 4 h and 10 dogs were sacrificed after 5 d. Temporary occlusion of coronary artery (from 30 min to 2 h) was produced by ligation. One group were controls and the other group were a hyperbaric group in which dogs breathed 100% oxygen at an absolute pressure of 2 atmospheres before and after release of coronary ligation. The macroscopic extent of ischaemic area was studied by using nitroblue tetrazolium and microscopic and ECG findings were examined. By breathing oxygen at high pressure immediately after reopening of occluded coronary blood flow, the ischaemic area was markedly reduced. In such cases, some myocardial muscles around the arterioles and sinusoids, even when these vessels existed in the ischaemic area, were kept in a viable state. The repair of necrotic myocardial muscles was promoted histologically. Serious arrhythmia, especially ventricular fibrillation, was also well suppressed, and the stable haemodynamic conditions were obtained during operative procedures. No harmful side effects of hyperbaric oxygen were observed. One of the most effective treatments of acute myocardial infarction involves reconstruction of the occluded coronary artery as soon as possible after the onset of myocardial infarction by using these advantages of hyperbaric oxygen.


Physiological disposition of verapamil in man. 14C-D,L-verapamil was administered intravenously (10 mg) and orally (80 mg) to five volunteer patients. Plasma concentrations of verapamil, which were determined by mass fragmentography, declined bi-exponentially with half-lives of the chi-phase ranging from 18 to 35 min and of the beta-phase from 170 to 440 min. The apparent volume of distribution ranged from 270 to 460 litre and plasma clearance from 730 to 1980 ml/min. Following oral administration absorption was almost complete as judged from the area under the curve (AUC) of 14C-activity and cumulative urinary excretion of 14C. After intravenous infusion of verapamil about 80% of the radioactivity administered could be recovered in urine and faeces within 5 d. Despite its almost complete absorption after oral administration verapamil was shown to undergo extensive first pass metabolism as the bioavailability was only 10 to 22%. Rapid biotransformation had occurred as only a small percentage of AUC of 14C was seen to correspond to unchanged verapamil after both intravenous and oral administration.


Nuclear transcripts larger than the cytoplasmic mRNAs are specified by segments of the adenovirus genome coding for early functions. Nuclear viral RNAs synthesized early in productive infection with adenovirus 2 were analyzed by hybridization to specific viral DNA fragments. Radioactive RNAs were subjected to electrophoresis in polyacrylamide gels containing 98% formamide, and the fractionated RNA was hybridized to specific DNA fragments generated by cleavage with endo R-Eco Ri or endo R-Sma l. The viral genes expressed early in infection are located in four different segments of the genome. When nuclear RNA was hybridized to DNA fragments representing each of these segments, discrete RNA size classes were detected. For each of these four regions of the genome, some of the discrete nuclear RNAs were larger than the cytoplasmic mRNAs. As calculated from electrophoretic mobility, the molecular weight of these nuclear RNA size classes ranged from 15% to several fold greater than that of the corresponding cytoplasmic mRNAs. Hybridization-inhibition experiments were performed to analyze the sequences present in one of these larger nuclear transcripts, a 22S RNA transcribed from the left-hand end of the genome. This 22S nuclear RNA was transcribed from the same strand as the viral mRNA, for it contained cytoplasmic sequences; it also contained sequences restricted to the nucleus. The larger nuclear RNAs may serve as precursors to the cytoplasmic viral mRNAs synthesized early in infection.


Induction of erythroid differentiation in vitro by purines and purine analogues. The effectiveness of purines and purine analogues as inducers  of erythroid differentiation in cultured murine erythroleukemia cells has been investigated. These cell lines have previously been shown to differentiate in vitro in response to dimethylsulfoxide (DMSO) and a number of other polar solvents. Two purine analogues, 6-thioguanine and 6-mercaptopurine, as well as the naturally occuring purine, purine, hypoxanthine, are shown to be extremely potent inducers. 6-Thioguanine is effective at a concentration of 0.06 mM, 750 fold lower than the DMSO concentration required for equivalent induction. 6-Mercaptopurine and hypoxanthine are effective inducers at a concentration of approximately 2 mM. Accumulation of globin mRNA was monitored during induction with purine inducers and shown to be similar in amount to globin mRNA levels reached in DMSO-induced cultures. Induction of differentiation by all three compounds follows a similar time course to induction with DMSO. All three compounds are potent inducers of HGPRT (hypoxanthine-guanine phosphoribosyltransferase)-negative cell lines; hence incorporation of purines into DNA is not required for induction of differentiation. Comparison of these compounds with other purines and purine analogues suggests a high degree of specificity in their interaction with a cellular target.


RNA synthesis in puff 2-48BC after experimental induction in Drosophila hydei. Puffs induced by treatment with vitamin B6 at locus 2-48BC in salivary gland cells of Drosophila hydei were isolated by micromanipulation, and the puff RNA was analyzed electrophoretically. The main RNA species migrated with a mobility of 40S. A similar RNA species was found in this puff after its induction by temperature treatment, except that in this case material with a higher mobility was also found, presumably due to the presence of growing chains. No evidence was found for polyadenylation of the RNA contained within the puff. The RNA profile of nucleoplasm of cells from glands either treated with vitamin B6 or subjected to a temperature shock also displayed a prominent peak with a mobility of 40S which was absent in extracts of nucleoplasm from control glands.


The expression of differentiated functions in somatic cell hybrids: retention and activation of C3 production. Rat and mouse fibroblasts which do not secrete the third component of complement (or secrete it at a very low rate), were respectively hybridized with mouse and rat hepatoma cells which actively produce C3. These interspecific hybrids actively secrete both mouse and rat C3. The production of C3 coded by the hepatoma cell genome is thus retained, but in addition, the secretion of C3 coded by the fibroblast genome is activated. On the other hand, intraspecific hybrids have been isolated from normal diploid C3 producing cells and non-producing cells. These cell hybrids also retain the capacity to secret C3. Retention of C3 production thus does not appear to be peculiar to hybrides derived from hepatoma cells.


Cultures of normal human mammary cells.

Cultures of normal human mammary cells. This paper describes a method of obtaining epithelial cells from large quantities of normal human breast tissue and the response of these cells in culture to lactogenic hormones. Suspensions of single cells and clusters of cells resembling normal ductal and alveolar structures were obtained by mechanical disaggregation and subsequent (3h) incubation of tissue fragments in 0.5 mg/ml collagenase. Cells rapidly attached to glass or plastic surfaces within 48 h and grew to form large colonies which maintained their epithelial appearance throughout 2 months of observation. Cell cycling as monitored by DNA synthesis was enhanced by insulin, hydrocortisone, or ovine prolactin (in concentrations of 5.0mug/ml each) at respectively 2,3 and 5 days of incubation. These results were observed in cultures derived from 3 premenopause samples of mammary tissue maintained in medium with 1% fetal calf serum. Prolactin at a concentration of 5 mug/ml induced phosphoprotein synthesis 8-fold over control values. In addition, prolactin induced morphological changes in cells including the development of distended endoplasmic reticulum, large microvilli, and the deposition of glycogen granules. These initial results led to the tentative conclusion that prolactin was sufficient to initiate some of the characteristics in cultured cells normally associated with lactating tissues.