Variation of forearm, hand, and finger blood flow indices with ambient temperature. Impedance plethysmography was used to measure bilateral blood flow in the forearms, hands, and digits of seven healthy male subjects. The objectives were to determine the degree of symmetry between right and left body segments and to quantify the extent of vasomotor change at ambient temperatures ranging from 10 degrees to 46 degrees C. The mean forearm blood flow index ranged from 3.9 ml/(100 ml . min) at 10 degrees C to 7.5 ml/(100 ml . min) at 46 degrees C. Mean hand blood flow indices increased from 1.6 ml/(100 ml . min) at 10 degrees C to 10.7 ml (100 ml . min) at 46 degrees C. Finger blood flow indices in the proximal and medial phalanges of the five digits ranged from less than 1.0 ml/(100 ml . min) at 10 degrees C to over 27.0 ml/(100 ml . min) at 46 degrees C. Although each subject demonstrated variations in right and left side blood flow during any selected experiment the grouped mean flow indices exhibited no statistically significant side dominance. These experiments quantify the vasomotor alterations in the upper extremities and indicate that the more distal body segments exhibit relatively larger vasomotor responses to changes in ambient temperature than do the more proximal body segments.


Optokinetic motion sickness: continuous head movements attenuate the visual induction of apparent self-rotation and symptoms of motion sickness. Symptoms of motion sickness are sometimes experienced during exposure to optokinetic stimulation. Two experimetns were performed to compare the symptoms of motion sickness  elicited when subjects were exposed to incremental changes in optokinetic stimulation while sitting passively and while continuously executing shoulder-to-shoulder head movements. In the first experiment, a fixed head-movement frequency (20 cpm) was used, wheras in the second the subjects varied the frequency of their head movements in order to maintain suppression of illusory self-rotation. In both experiments, subjects in the head-moving condition had fewer and less severe symptoms of motion sickness and experienced illusory self-rotation after longer exposure times and at higher optokinetic velocities than in the head-stationary condition. Subjects in th- head-movement condition of the second experiment increased the frequency of their head movements as the velocity of optokinetic stimulation increased. The symptoms of motion sickness elicited during optokinetic stimulation tended to be dizziness, headache, eye-strain, and stomach awareness appearing in no fixed order. The pattern and constellation of symptoms are unlike those elicited by vestibular stimulation.


Responses to temperate, cold, and hot environments and the effect of physical training. Ten young men underwent several tests before and after a training program: a bicycle ergometer test and 60 min of moderate exercise performed at a temperate 24 degrees C; the same work load performed in heat (40.0 degrees C DB, 30.4 degrees C WB) for 3 h; and cold (10 degrees C) exposure for 60 min. Training consisted of 13 1-h sessions of hard, strenuous, and exhaustive work performed in temperate conditions four times a week. Training resulted in substantial decreases in heart rate and rectal temperature responses to exercise in temperate, minor increases in hot, and no significant changes in cold conditions. Subjects who showed good responses to heat, also showed good responses at 24 degrees C, and poor compensatory responses to cold, which were indicated by relatively low heat production and rectal temperature values, and relatively high body heat loss and extremities temperature values. Subjects who showed poor heat tolerance also showed poor responses in temperate and good compensatory responses in cold conditions. Positive correlation coefficients were found between rectal temperatures in the three environments, and between heart rate and sweat rate responses in temperate and hot conditions. The results indicated that moderately severe training causes minor tolerance improvements in heat and no changes in cold, and that responses in temperate, cold, and hot environments are interdependent.


Direct and indirect ophthalmoscopy for a more accurate baseline evaluation in aircrew members. The currently required Federal Aviation Agency visual evaluation for commercial and airline pilots often does not detect quiescent retinal disease, unless there is a specific history or a current change in visual acuity which dictates the need for a dilated ophthalmoscopic evaluation. Statistics indicate that there may be a significant number of undetected retinal changes which can cause sudden and irreversible alterations in visual acuity during an airman's career. The requirements for an ophthalmoscopic examination should include, at the time of entry as an aircrew member into the aviation industry, a dilated fundus examination by the binocular indirect and direct ophthalmoscopic methods. In addition, documentary photography, visual fields, and other specific studies as indicated for these patients would be accomplished. These studies should be required by both the Federal Aviation Agency and the military services just as baseline ECG's chest films, SMA 12, and other laboratory studies are utilized.


Multicenter evaluation of synovectomy in the treatment of rheumatoid arthritis. Report of results at the end of three years. A controlled multicenter evaluation of synovectomy in the treatment of rheumatoid arthritis followed 59 patients for 3 years. Finger joints and knees were studied. After 1 year synovectomized joints were better than controls in several features. After 3 years these differences had disappeared in the fingers, but swelling remained less in synovectomized knees. Recurrences were as frequent in synovectomized as in control finger joints, but were slightly less frequent in synovectomized than in control knees. Radiologic bony erosions in fingers and knees of both groups progressed equally during the 3 years.


Chalky joint effusion. The result of massive synovial deposition of calcium apatite in progressive systemic sclerosis. A patient with long-standing progressive systemic sclerosis developed calcification of the suprapatellar pouch. Chalky white synovial fluid from the knee joint contained many crystals that were characterized as apatite by X-ray diffraction. In this case massive amounts of cystals were presumably shed into the fluid from the heavily calcified synovium.


Antigen-induced arthritis in mice. I. Induction of arthritis in various strains of mice. Antigen-induced arthritis was established in the mouse by immunizaiton with methylated bovine serum albumin (mBSA) in complete Freund's adjuvant with B  pertusis vaccine. The knee joint was injected after 21 days with mBSA in saline. The arthritis was chronic, antigen-specific, and T-cell dependent in hypothmic nu/nu mice. C57BL and balb/c mice were susceptible, whereas CBA mice were relatively resistant. Susceptibility was dominant; one gene was loosely linked to the "b" allele of the H-2 complex of C57BL mice.


Permeability of inner and outer layers of rat and rabbit aortic wall. Two new microscopic test with trypan blue. Two new permeability tests are described for use with intravenously injected trypan blue. One depends on the demonstration of trypan blue by its specific red fluorescence in green light at 570 nm, while the other is a surface microscopy technique at low magnification, using illumination from a fibre-optics light source. The routes of entry of the trypan blue-albumin complex into the rat and rabbit aorta appear to be from both the inner and outer surfaces. Over the period 1/4-24 h after injection of the dye, more entered from the outer surface than the inner surface in the rat aorta and rather more in the rabbit thoracic aorta. The arch of the rabbit aorta showed in general rather greater entry from the inner surface. Trypan blue that has entered the aortic wall is partly taken up by the elastic lamellae. Elastic competes successfully for the dye and captures it from the trypan blue-albumin complex; this uptake is blocked by deamination with nitrous acid.


Synthesis and removal of different cholesterol esters by aortic smooth muscle cells in culture. The incorporation of 3H-labelled oleic acid and of 14C-labelled linoleic acid into phospholipid, triglyceride and cholesterol ester in smooth muscle cells grown in incubation medium supplemented with either 5% normal or 5% hyperlipemic serum has been studied. Both fatty acids were incorporated into cholesterol esters to a greater extent when cells grown in incubation medium containing hyperlipemic serum. Oleic acid was incorporated into cholesterol esters in preference to linoleic acid. The addition of hyperlipemic serum to the incubation medium did not increase the incorporation fo either 3H-labelled oleic acid or of 14C-labelled linoleic acid into phospholipid or triglyceride. The removal of labelled lipid fractions has also been followed for four days in cells pulse labelled for 24 hours with 3H-labelled oleic acid and 14C-labelled linoleic acid. Both 3H- and 14C-labelled cholesterol esters were removed more rapidly when the smooth muscle cells were grown in medium containing normal serum than in medium containing hyperlipemic serum. The removal of both phospholipid and triglyceride was similar in normal and hyperlipemic serum. Comparison of the 3H/14C ratio indicated that the cholesterol oleate and cholesterol linoleate were removed at similar rates.


Uptake and metabolism of 3H-fatty acid labelled lecithin by normal and atherosclerotic intima in vivo and in vitro. Following the intravenous injection of tracer doses of 3H-fatty acid labelled lecithin into normally fed rabbits, minimal incorporation into serum cholesterol ester over a 6-hour period was observed. The labelled phospholipid was rapidly removed from the serum primarily by R.E. tissues with little uptake by the aorta. The uptake and metabolism of 3H-fatty acid labelled lecithin by explants of normal and cholesterol-fed rabbit aortas was also investigated in vitro for periods up to 8 days. Low uptake and incorporation into cholesterol ester was observed for the normal aortic explants. The labelled phospholid was taken up to a greater extent by explants from cholesterol-fed rabbit aortas and the fatty acid incorporated into both triglyceride and cholesterol ester in the explants. The incorporation of fatty acid into cholesterol ester from phospholipid was associated with hydrolysis of the phospholipid and re-incorporation of the fatty acid into cholesterol ester, rather than by the action of lecithin cholesterol acyl transferase.


Preventive trials and the "diet-heart" question: wait for results or act now. Is the relationship between coronary heart disease risk factors and the risk of disease causal? Direct evidence would have to come, of necessity, from preventive trials. However, the difficult nature of such trials carries the inherent danger of false negative results; even positive results may be open to challenge. With regard to demonstrating specifically a preventive effect of lowering serum lipid levels through nutritional changes alone, this evidence may never be forthcoming since ongoing trials intervene simultaneously on multiple factors; against this disadvantage, the strengths of such trials are reviewed. The view is stated and discussed that preventive action in the community is justified now, without waiting for the results of preventive trials, however important they will be eventually to help the development of optimally effective programs.


Pre- and post-natal development of lipoprotein lipase and hepatic triglyceride hydrolase activity in rat tissues. The ontogenic development of lipoprotein lipase and liver triglyceride hydrolase was studied in the rat. The enzyme activity measured in extrahepatic tissues fulfilled the criteria of lipoprotein lipase from the onset of measurable activity, i.e. it was inhibited by protamine and 1 M NaCl and showed requirement for serum and heparin for optimal activity. In the liver, measurable amounts of triglyceride hydrolase, active at pH 8.6 were detected 6 days prior to birth. However, till the fourth postnatal day about 50% of this activity was inhibited by NaCl and its sensitivity towards protamine was also higher than that of the enzyme in adult liver. Three patterns of development of enzymic activity were observed in extrahepatic tissues. In the lung, the lipoprotein activity reached the adult values one day prior to birth, while in the kidney only 30% of adult activity were found at birth. A linear increase of enzyme activity was observed in the heart; only 25% of adult activity were detected at birth and 100% were reached only 20 days after birth. The increase in lipoprotein lipase activity in the heart was accompanied by morphological differentiation of cardiocytes and by a progressive development of the capillary bed, which might be related to the pattern of development of enzyme activity in this organ. Adipose tissue lipoprotein lipase activity in inguinal fat fell from values 15 times than adult values between the 4th and 40th postnatal days. The enzyme activity in epididymal fat increased steeply between day 10 and 40, at which time it exceeded the adult values very considerably. These findings indicate that the regulation of the development of lipoprotein lipase activity in extrahepatic tissues is governed by local factors, which can differ even in the same type of tissue, as exemplified by the difference between inguinal and epididymal fat.


The special hemodynamics and metabolism in severe disturbances of peripheral blood flow and their treatment. Hemodynamic investigations were carried out in the various stages of a peripheral arterial occlusive disease, primarily using venous occlusion plethysmography, and simultaneous metabolic determinations were performed in groups of 60 patients. As a rule, there were poorly collateralized occlusions at several levels in the severe stages of the peripheral blood-flow disorders. Venous occlusion plethysmography shows a pathological profile of reactive hyperemia. The parameters of resting blood flow in the severe stages were not found to be lowered in any patient; their average value was higher than that of a healthy comparison group (60 limbs). The oxygen saturation in the femoral vein in almost equal to that in the femoral artery in the exercise period in these severe stages, i.e. no oxygen utilization takes place in the periphery. While a rise in various enzyme activities could be observed after ergometric exercise in patients in stage II according to Fontaine, this is not possible in stages III and IV. There were no significant differences in measurements of the arteriovenous lactate-pyruvate difference, the pO2, the pH, the standard bicarbonate the hematocrit and the viscometry. In the severe stages, there is thus a situation of anaerobic metabolism and an accumulation of acid metabolites. An attempt is made to explain why a systematically perfomed vasodilatation has no positive effect but can lead to undesired side-effects such as a borrowing-lending phenomenon. Intra-arterial vasodilatation with short-acting vasodilators can only lead to a necessary blood-flow when the vascular occlusion is compensated by a well collateralized cirulation. In the severe stages, a fall in the perfusion pressure and blocking of the terminal blood vessel is possible. In advanced stages of a blood flow disorder it is therefore necessar during the first infusion to demonstrate by venous plethymography a quantitative increase in blood flow and to exclude a local peripheral steal effect. Various other possibilities of therapy are indicated.


The relationship of abnormal circulating insulin levels to atherosclerosis. The evidence linking insulin with atherosclerosis can be divided into two parts. First, there is evidence that a proportion of subjects who have atherosclerosis or who are at risk of developing atherosclerosis hav elevated circulating insulin levels. The high insulin levels may be associated with another metabolic abnormality such as obesity, hypertriglyceridaemia, uraemia or consumption of oral contraceptives, may be inappropriate to the blood sugar levels as in mild diabetes, or may be of exogenous origin as in insulin-treated diabetics. The tissues of these subjects are exposed to high concentrations of insulin, and it seems reasonable to suggest that elevated insulin levels may have pathological effects on these tissues. Secondly, there is increasing evidence that the arterial wall is an insulin sensitive tissue. Exposure of arterial tissue to insulin results in proliferation of smooth muscle cells, inhibition of lipolysis, and synthesis of cholesterol, phospholipid and triglyceride. Chronic exposure to high concentrations of insulin results in the development of lipid filled lesions similar to those of early atherosclerosis. Thus, insulin has the ability to promote changes in the artery which, in the long term, may progress to atherosclerosis. The two lines of evidence together suggest that high levels of circulating insulin may have a role in the development of atherosclerosis.


Comparison of hypocholesterolemic activity for cyclic analogs of clofibrate in normolipemic rats. Chronic administration of ethyl 2-methyl-2(4-chlorophenoxy)-propionate [clofibrate,  CPIB], ethyl 6-cyclohexylchroman-2carboxylate, and ethyl 6-phenylchroman-2-carboxylate to normolipemic rats, in vivo, reduced serum cholesterol levels and inhibitid the activiry of hepatic 3-hydroxy-3methyl-glutaryl Coenzyme A. Only clofibrate was found to lower liver cholesterol content after pretreatment for 4 or 18 days. The cyclic analogs, ethyl 6-cholorochromone-2-carboxylate and 9-chloro-2,3-dihydro-5H-1,4-dioxepino [6,5-b] benzofuran were inaffective as cholesterol lowering agents in normolipemic rats. These findings indicate that appropriate modification of clofibrate can lead to the development of compounds which are selective and equally effective to clofibrate as potential hypocholesterolemic agents. Results obtained in these studies are also discussed in terms of the known structural requirements of biological activity for this series of cyclic analogs in the Triton WR-1339 hyperlipemic rat model and modes of action of the parent compound.


Plasma renin activity in men with relation to the presence of ischaemic heart disease. Plasma renin activity (PRA) stimulated by upright posture was measured, in 300 men aged 45-64 years, by a radio-immunoassay of angiotensin I. The subjucts examined were divided into six groups, comparable in mean age, each containing 50 subjects: group 1, normotensives without manifest atherosclerosis; group 2, normotensives with angina pectoris definite; group 3, normotensives with a history of a transmural myocardial infarction; groups 4 to 6, patients with benign essential hypertension, without manifest atherosclerosis in group 4, with angina pectoris in group 5 and with a history of myocardial infarction in group 6. Significant differences in mean PRA were found between corresponding groups of hypertensives and normotensives, the values in hypertensives being lower. The precentage of low renin values  was higher in hypertensives with ischaemic heart disease than in other groups.  An analysis of 3-year cardiovascular mortality revealed no significant difference in mortality due to ischaemic heart disease between high-renin and low renin sub-groups.


The effects of supplemental dietary cholesterol and exercise on blood cholesterol and atherosclerosis in the goat. The objective of this experiment was to determine the effects of supplemental dietary cholesterol and treadmill exercise on blood plasma cholesterol and development of atherosclerosis in young goats. Eighteen two-week-old goats, assigned to four groups for 22 weeks, were fed 100 g whole milk and, after 14 weeks, 50 g corn and cob meal daily/kg body weight. The four groups received, respectively, 250, 175, 100 and 25 mg cholesterol/kg body weight daily in the milk. From week 10 to week 22 of the experiment half of the goats in each group were exercised on a motorized treadmill at a rate of 6.4 km/h for 15 min daily, five days per week. Addition of cholesterol to the diet increased the mean plasma cholesterol in each of the four groups, but between-group comparisons were statistically significant for only the 250-mg group versus the 25-mg group. Exercise had no effect on plasma  cholesterol. Analysis of serial blood samples of 16 of the goats during one day of the experiment indicated no significant diurnal variations. Six males, three exercised and three nonexercised, were sacrificed; all had extensive aortic sudanophilia. Histological preparations from sudanophilic areas of all aortas showed areas of intimal thickening composed of foam cells. These same areas stained strongly for lipid with Oil Red 0. No histological evidence of calcium deposition or fibrous plaques was found.


Incidence of congenital heart disease in Blackpool 1957-1971. In the 15 years from 1957 to 1971, there were 57 979 births in a circumscribed population in North-West England. The requirements for a valid survey of the incidence of congenital heart disease were largely met. The necropsy rate for all stillbirths and neonatal deaths in the population was 93 per cent. Diagnosis of the type of heart lesion was by necropsy in 50 per cent, cardiac catheterisation in 19 per cent, operation in 5 per cent, and clinical observations only in 26 per cent. The incidence of congenital heart disease was 6-8 per 1000 total births and 5-9 per 1000 live births. The overall incidence has remained unchanged over the 15 years, but there were trends suggesting an increase in the incidence of uncomplicated ventricular septal defect and of endocardial cushion defect and a decrease in the incidence of ventricular septal defect with right ventricular outflow obstruction and of hypoplastic left heart. We have observed seasonal variations in total incidence and in incidence of some common individual lesions.


Ventricular haemangioendothelioma diagnosed in life. A case of right ventricular outflow obstruction is described in which the electrocardiogram and chest x-ray film suggested a cardiac tumour. Cardiac catheterisation confirmed outflow obstruction and angiography showed a tumour. At operation a haemangioendothelioma was found.


Syphilitic ostial occlusion.

Syphilitic ostial occlusion. A patient presenting the picture of congestive cardiomyopathy was found to have syphilitic obliteration of the left coronary artery. Aortic regurgitation, angina, and myocardial infarction were notably absent.


A new cell line from a human chondrosarcoma. Morphological and growth characteristics are described of a rapidly growing cell line with epithelioid and giant-cell characteristics derived from a chondrosarcoma in a male patient 65 years of age. This cell line is of considerable interest because in these cells cross-reacting antigens with known animal oncorna-viruses are present. Biochemically, the cells contain particles with a density of 1-16 with "cores" of density 1'23 associated with a reverse-transcriptase-like enzyme and with 70S RNA. Occasionally, virus-like particles were demonstrated by electron microscope in material derived from the culture medium.


Effect of inhaled plutonium dioxide on development of urethan-induced  pulmonary adenomas. Mice were exposed to plutonium dioxide (PuO2) aerosols 2 weeks before or after urethan injection. Both exposures reduced the number and size of adenomas. The incidence of arrested metaphases showed no consistently significant differences between plutonium-exposed and mock-exposed animals. The results are discussed in relation to recent electron microscopic evidence of degenerative changes in the type II epithelial cells of the mouse lung following PuO2 inhalation. It is concluded that damage at the cellular level may account for the observed reduction in growth of pulmonary adenomas in mice whose lungs contained plutonium particles.


Effects of C. parvum on growth and induction of intracerebral tumours in mice. An investigation was made into the effect of Corynebacterium parvum therapy on cerebral tumours in mice. I.v. C. parvum caused a slight but significant increase in the survival of BALB/c mice injected intracerebrally (i.c.) with not more than 50 Meth A cells. C. parvum was most effective if given on the same day or 5 days after tumour. If this interval was increased there was no effect. Multiple i.v. injections were no more effective than a single dose. I.v. C. parvum had no influence on the survival of C57BL mice injected i.c. with Lewis tumour cells, and had little effect on the induction of i.c. or s.c. tumours by methylcholanthrene. It was concluded that C. parvum therapy was of little use in the treatment of cerebral tumour in mice. The clinical implications of these findings are discussed.


Localization of malanin pigmentation in the skin with Wood's lamp. Examination of the skin or hair with Wood's lamp has long been used to aid in the clinical diagnosis of some cutaneous disorders. A new observation reported here illustrates that Wood's light can be used to determine the depth of melanin pigmentation in the skin: contrast in epidermal pigmentation is increased while contrast in dermal pigmentation is decreased under Wood's lamp illumination compared to ambient visible light. The principles underlying this phenomenon are discussed. Use of the Wood's lamp to localize abnormal melanin pigmentation in the skin can be a guide to clinical diagnosis of hypermelanosis.


A histological study of normal vulval skin from infancy to old age. A number of histological parameters in clinically normal skin from the vulva throughout the female life span were studied. These are compared and evaluated to establish a base line to assist with the interpretation of diseased vulval skin.


Multiple lesions of the lip exhibiting the "Jadassohn phenomenon". Review of the literature and report of a case. The clinical and microscopic features of multiple, superficial lesions of the lip and adjacent mucosa in an elderly woman are reported. The lesions had been present for many years apparently without symptoms. A histologically confusing picture gave rise to diagnostic problems. Characteristic nests of basaloid cells were present at all levels of the acanthotic epithelium together with microcysts, horn pearls and squamous cell eddies. Review of the literature revealed no previous report on similar lesions of the oral mucosa and it is suggested that the lesions closely coincide with the Jadassohn type of intra-epidermal epithelioma.


Toxic epidermal necrolysis in two patients with pustular psoriasis. Two cases of toxic epidermal necrolysis are described in patients suffering from pustular psoriasis. In one of these cases there were no recognized predisposing causes. In the other, although there were alternative possibilities, we consider that the most likely cause of the toxic epidermal necrolysis was pustular psoriasis.


Monocyte recruitment in tuberculosis and sarcoidosis. Monocytopoiesis and blood monocytes were investigated in nine patients with active tuberculosis and in six patients with active sarcoidosis in order to obtain information on monocyte consumption in these two types of granuloma. All patients with tuberculosis demonstrated a marked increase in proliferation activity of monocytopoiesis and premature monocyte marrow release. These changes indicate a high monocyte consumption which probably is caused by a high macrophage death rate due to the high macrophage-toxicity of tubercle bacilli. Thus, tuberculous lesions are an example of a "high turnover granuloma". In sarcoidosis monocytopoiesis showed no significant deviations from the normal. This indicates a low macrophage turnover or "low turnover granuloma". Thus, any hypothetical agent assumed to be involved in the pathogenesis of sarcoidosis would have to possess low macrophage-toxicity.


Haematologic profile of natural populations: red cell parameters. Age-sex specific reference values and frequency distribution of erythrocyte parameters were derived from a representative sample of a natural population consisting of 1094 men and 1284 women between 10 and 96 years of age. Some differences have been found between previously reported reference values and those presented here, the importance of which lies not so much in their magnitude as in the fact that the latter are based upon a probability sample of a natural population unselected as to disease and are presented as percentile distributions by age and sex rather than as the more frequently used normal distribution. These results are intended for use as reference data both for clinical and for population estimates of haematological parameters.


Neutrophil accumulation in vivo following the administration of chemotactic factors. Intravenously administered 51Cr-labelled homologous neutrophils accumulated at guinea-pig skin sites prepared by intradermal injection of factors derived from complement-activated serum, possessing in vitro chemotactic activity. There was a strong correlation between the in vitro potency of Sephadex G-100 fractions of the activated serum and their ability to evoke neutrophil accumulation in vivo. These experiments suggest that agents which are chemotactic for neutrophils in vitro also induce the localization of this cell in vivo.


Benzene inhibition of in vitro rabbit reticulocyte haem synthesis at delta aminolaevulinic acid synthetase: reversal of benzene toxicity by pyridoxine. Benzene (0.113 M) inhibited haem and protein synthesis in rabbit reticulocytes. This inhibition of haem synthesis was found when L-2-[14C]-glycine was used as the radioactive precursor. However, when 4-[14C]delta-aminolaevulinic acid (ALA) was used, there was no significant inhibition. Since ALA measures the haem synthetic pathway beyond the enzyme delta-aminolaevulinic acid synthetase (ALA synthetase), these results suggest that benzene inhibits haem synthesis at or before ALA synthetase. This was confirmed by demonstrating that 1 mM ALA both protected against and reversed the benzene inhibition of reticulocyte protein synthesis. In addition, 1 mM pyridoxine both protected against and reversed the benzene inhibition of reticulocyte protein synthesis. In addition, ImM pyridoxine both protected against and reversed the benzene inhibition of reticulocyte haem and protein synthesis. These results indicate that benzene (or a metabolite) either competes with pyridoxal phosphate at ALA synthetase or competes with pyridoxine for pyridoxal phosphokinase. These results are discussed in terms of their implications for the possible roles of ALA synthetase and the haemin-controlled repressor in benzene-induced aplastic anaemia.


Haemoglobin Radcliffe (alpha2beta299(Gi)Ala): a high oxygen-affinity variant causing familial polycythaemia. Three members of an Oxfordshire family have polycythaemia. In each case their whole-blood oxygen affinity is increased. This is due to a previously undescribed haemoglobin variant which has been named haemoglobin Radcliffe (alpha2beta299(Gl)Ala). In addition to having a high oxygen affinity haemoglobin Radcliffe shows virtually no haem-haem interaction and a diminished Bohr effect. It is synthesized at the same rate and is as stable as haemoglobin A. X-ray analysis indicates that crystals of deoxyhaemoglobin Radcliffe are isomorphous with those of deoxyhaemoglobin A. Solutions of haemoglobin Radcliffe were also studied by high-resolution proton nuclear magnetic resonance spectroscopy. The structure/function relationships of haemoglobin Radcliffe are discussed in the light of these studies.


Reversal of ineffective erythropoiesis in pernicious anaemia following vitamin B12 therapy. Ineffective erythropoiesis was quantitated in a series of patients with pernicious anaemia at different times in relation to vitamin B12 therapy by measuring the incorporation of [15N]delta aminolaevulinic acid and [15N]glycine into early labelled bilirubin. Prior to therapy ineffective erythropoiesis was grossly increased but this was reversed within 24 h of giving vitamin B12, suggesting that most of the existing megaloblasts are enabled to mature into circulating red cells.


Defective platelet lipid peroxidation in myeloproliferative disorders: a possible defect of prostaglandin synthesis. Platelet lipid peroxidation induced by the sulphydryl blocking reagent n-ethylmaleimide [NEM] was measured by production of malonyldialdehyde [MDA]. Lipid peroxidation was decreased in five of 10 patients with proven myeloproliferative disease. The low levels of lipid peroxidation correlated well with in vitro and in vivo tests of platelet function and the bleeding and bruising problems. The reduction in lipid peroxidation with NEM may be due to one or more of the following: [i] an aged platelet population; [ii] an abnormal cell line; [iii] a reduction of the cyclo-oxygenase enzyme activity in the platelet prostaglandin synthetic pathway; [iv] a reduction of the substrate for platelet prostaglandin synthesis, Arachidonic acid; and [v] an abnormality of phospholipase A activation.


Inherited sialoglycoprotein deficiencies in human erythrocytes of type En[a-]. We have investigated the membranes of erythrocytes from a family in which there is a genetic defect [previously described as the En[a-] condition[ resulting in the loss of the major erythrocyte sialoglycoprotein [PAS-i]. The results show that two different types of sialoglycoprotein deficiency can be distinguished within this family. We suggest that the En[a-] group of variants is more appropriately described as a class of sialoglycoprotein deficient erythrocytes. Using a new technique it is shown that the blood group M antigen of normal erythrocytes is found only on the erythrocyte sialoglycoprotein while in this family the M antigen is found on membrane components other than the sialoglycoprotein. Our results suggest that the amino acid sequence of the sialoglycoprotein is important in defining the difference between the blood group M and N antigens in normal erythrocytes.


Immunoglobulin synthesis in hairy cell leukaemia. In vitro studies were performed with leukaemic cells from two patients with hairy cell leukaemia in order to define the nature and kinetics of immunoglobulin synthesis by the neoplastic cells. Both patients had clinically and morphologically well-defined disease and their cells contained abundant tartrate-resistant acid phosphatase. One patient had associated macroglobulinaemia. The hairy cells had B-lymphocyte characteristics as determined by fluorescent immunoglobulin staining and surface receptor properties. They synthesized monoclonal IgM and IgG respectively in vitro. The kinetics of immunoglobulin synthesis were different in cells from the two patients as measured by equilibration time, intracellular degradation, and secretion. Permanent cell lines were established with cells from these patients. The lines grow as typical B-lymphoblastoid cultures and continue to produce tartrate-resistant acid phosphatase and immunoglobulin. These studies unequivocally demonstrate the B-lymphocyte nature of the hairy cells in these patients and provide evidence for their clonal origin both in terms of immunoglobulin and enzyme synthesis.


Altered haem--haem interaction and tissue-oxygen supply: a theoretical analysis. On the basis of its assumed invariance, the haem--haem interaction or shape of the oxyhaemoglobin dissociation curve is frequently ignored when considering the potential effect of haemoglobin-oxygen affinity (as characterized by the P50) on oxygen transport. This report reviews some experimental evidence with respect to this assumption and demonstrates the potential effect of altered haem--haem interaction on tissue-oxygen supply as distinguished from the effect of P50 or position change. Theoretically, haem--haem interaction values in the range that has been reported for apparently healthy human subjects can substantially modify oxygen transport to tissues, in some cases equalling or exceeding the effect from a similar range of P50 values. It is not presently clear to what extent this range may represent true variation as opposed to random technical error, but the possible implications for oxygen transport indicate that a more systematic evaluation of the problem is desirable.


Pathophysiological changes in serum erythropoiesis stimulating activity. The mouse fetal liver cell bioassay has been used to study serum levels of erythropoiesis stimulating activity (ESA) in patients with a variety of haematological disturbances. The results indicate that this assay system provides a quantitative measurement of ESA, the concentration of which varies predictably in a manner which is consistent with it having a function as an in vivo regulator of erythropoiesis. Dose-response curves for serum ESA show highly significant parallelism with those obtained from sheep plasma erythropoietin and the International Reference Preparation of human urinary erythropoietin.


Factor XI and platelets: Evidence that platelets contain only minimal factor XI activity and antigen. Factor-XI activity of platelets has been studied in platelet-rich plasmas and isolated platelet suspensions. Fresh platelets in both environments had little or no measurable factor-XI activity. Frozen and thawed platelet-rich normal plasma had markedly elevated apparent factor-XI activity and factor-XI activity as compared to platelet-poor plasma. Frozen and thawed platelet-rich and platelet-poor normal plasmas had equivalent factor-XI antigen. Platelets isolated from normal blood and from factor-XI deficient blood had the same small amounts of apparent factor-XI activity, which increased slightly on freezing and thawing. The data indicates that minimal factor XI is associated with the platelet. The markedly elevated apparent factor-XI activity of frozen and thawed platelet-rich plasma is shown to reflect the interaction of a platelet activator with plasma clotting factors to produce a later activated-clotting-intermediate.


Isolation and characterization of an actin-like protein from membranes of human red cells. The existence of an actin-like protein in human red-cell membranes has been confirmed. The protein was extracted from acetone-treated ghosts and purified by (NH4)2SO4 fractionation. The protein undergoes G-F transformation and forms filaments in the presence of 0.1 M KCl. The filaments can be "decorated" by muscle heavy meromyosin. The protein has the same molecular weight as muscle actin and interacts with muscle myosin. All these properties show that the protein closely resembles muscle actin.


Depression of the platelet count after inoculation of mice with L1210 or L5178Y cells. The inoculation of L1210 or L5178Y leukaemia cells decreased the platelet count of the recipient mouse before extensive infiltration of marrow was expected by either tumour line. The decrease was more pronounced after intravenous inoculation of L1210 leukaemia than after intraperitoneal inoculation. Inoculation of L5178Y cells by both routes caused an initial decrease but the count recovered to 80% of normal before becoming markedly depressed prior to death of the host. Implantation of diffusion chambers containing L1210 cells also decreased the platelet count, which later returned to normal.


The effect of altering thyroid hormone concentrations on plasma gonadotrophins in postmenopausal women. Plasma gonadotrophin levels were measured at frequent intervals in six postmenopausal thyrotoxic women treated with carbimazole. All patients showed an increasing follicle stimulating hormone/luteinizing hormone (FSH/LH) ratio as the thyroid hormone concentrations fell to normal and then into the hypothyroid range. Although the increased FSH/LH ratio was primarily due to a rise of FSH, two patients had a rise of both gonadotrophins. It is suggested that menstrual irregularities occurring in premenopausal women with disorders of thyroid metabolism may be related to similar changes.


Ripening of the cervix with extra-amniotic prostaglandin E2 in viscous gel before induction of labour. A metohod of extra-amniotic prostaglandin administration to achieve ripening of the cervix as a preliminary to induction of labour is described. Compared with a control group the method achieved a 25 per cent reduction in the length of the induced labour, a fall in the incidence of maternal pyrexia and fetal distress, an improvement in neonatal condition of the infants and a reduction in the need to resort to Caesarean section.


Maternal plasma glucose and free fatty acid concentrations related to infant birth weight. A new and specific method is used for the measurement of plasma free fatty acids (FFA) in pregnancy. Fasting plasma FFA and glucose concentrations were measured serially in twenty normal and twelve overweight women in pregnancy and related to infant birth weight. Large variation between individuals was noted for FFA and no change was found with advancing gestation. Studies of the day-to-day variation in the same women at 20 and 36 weeks gestation showed wide variations in concentrations. No significant change in glucose concentration was noted with advancing gestation, but postnatal values were significantly higher. Small variability of glucose concentrations was noted in both the serial and day-to-day studies. In normal-weight women a positive correlation between the observed birth weight of their infants and the fasting plasma glucose levels was found at 20 and 40 weeks and also with the mean of 5 levels measured in pregnancy. No similar correlation was found for the group of overweight women.


Arterial blood gases in pre-eclampsia.

Arterial blood gases in pre-eclampsia. Arterial blood gases, physiological dead space and percentage pulmonary venous admixture (physiological shunt) were measured in 31 patients with proteinuric pre-eclampsia. There was no difference in pulmonary function in 22 patients with moderate pre-eclampsia when compared with normal pregnancy, but in 9 patients with severe pre-eclampsia, there was a significant increase in alveolar-to-arterial PO2 difference and physiological shunt, indicating a degree of pulmonary ventilation/perfusion imbalance. There was no accompanying hypoxaemia, suggesting that the impairment was of no clinical importance.


Amniotic fluid lecithin/sphingomyelin ratio in pregnancy complicated by diabetes. The lecithin/sphingomyelin (L/S) ratio in the amniotic fluid of 57 patients attending an antenatal diabetic clinic was determined. The L/S ratio in Class A, B and C patients (White, 1965) showed a similar rise with gestation to nondiabetic patients; 89 per cent in this group had an L/S ratio above 2 : 1, but of these 27 per cent developed mild or moderate respiratory distress syndrome (RDS). In Class D, F and R diabetic patients there was a little correlation of the L/S ratio with gestational age and in this group 55 per cent of the infants developed some form of RDS. The value of measuring amniotic fluid L/S ratios in the diabetic pregnancy is questioned.


Excretion urography before and after surgical treatment of procidentia. Excretion urography in 18 patients with procidentia confirmed the presence of a significant incidence of urinary tract obstruction and its relief following corrective surgery.


Treatment of recurrent pregnancy hypertension by prophylactic anticoagulation. A patient is described who had two pregnancies complicated by severe pre-eclampsia in the second trimester which resulted in intrauterine death. During the third pregnancy anticoagulant therapy with warfarin was instituted at 14 weeks gestation and a healthy female infant was delivered at 38 weeks gestation.


Microfilaria malayi in uveitis: case report. A case of uveitis with microfilariae in the anterior chamber is reported. The organism was recovered from the aqueous and identified as Brugia malayi.


Behaviour of the iris vasculature in central retinal vein occlusion: a fluorescein angiographic study of the vascular response of the retina and the iris. The findings in iris fluorescein angiograms of 48 eyes with central retinal vein occlusion (CRVO) were correlated with the predominant retinal vascular response. In 24 eyes with the non-ischaemic type of CRVO there were no or only minimal iris vessel changes, whereas in all 24 eyes with ischaemic type of CRVO there was iris vessel dilatation and leakage with or without neovascularisation. These findings support the hypothesis that hypoxic retina may be a cause of rubeosis iridis in CRVO.


Trabeculotomy in pseudoexfoliation of the lens capsule. Trabeculotomy was performed on 12 eyes of 11 patients with glaucoma associated with pseudoexfoliation of the lens capsule. Pressure was lowered in all eyes, in 10 eyes to 15 mmHg or less, 7 with no medication but 3 with medication. A subconjunctival bleb was present in 5 patients but was not necessary for a good result. These results suggest that the raised intraocular pressure which often occurs in pseudoexfoliation of the lens capsule is due to interference with flow of aqueous across the trabecular meshwork. Trabeculotomy seems a safe and effective operation in glaucoma associated with pseudoexfoliation of the lens capsule.


Episcleral traumatic conjunctival inclusion cyst. A hugh episcleral traumatic conjunctival inclusion cyst enveloping almost the whole eye, which appeared in the left phthisical eye of a young woman who sustained a perforating ocular injury a few years before enucleation, is described.


Nitrogen retention in rats fed on diets enriched with arginine and glycine. 2. Effect of diethyl ether anaesthesia on N retention. 1. Adult rats were subjected to a brief period of diethyl ether anaesthesia and were given diets with 200 or 100 g casein/kg with or without arginine plus glycine supplementation in the post-anaesthesia period. Nitrogen retention was measured as well as liver protein content and liver and muscle transaminase activities (L-aspartate aminotransferase (GOT), (EC 2.6.1.1), and L-alanine aminotransferase (GPT)(EC 2.6.1.2). 2. Results demonstrated that anaesthesia-stressed rats consuming the high-protein diet with supplemental arginine and glycine retained twice as much N as did rats given the diet with 200 g casein/kg alone, for the first 5 d post-anaesthesia. 3. Anaesthesia-stressed animals consuming the diets with 100 g casein/kg with or without arginine plus glycine supplementation did not differ from each other in N retention. 4. Liver protein content increased after anaesthesia in rats given the high-protein diets; liver transaminase activity increased, whereas muscle transaminase activity decreased, in animals consuming the high protein diets. 5. Possible mechanisms to account for these results are discussed.


The effect of wheat fibre on plasma lipids and faecal steroid excretion in man. 1. The effect of isoenergetic substitution of bran and wholemeal bread for white flour and white bread was investigated in six healthy volunteers in a metabolic unit. 2. A 21 d high-fibre experimental period was preceded and followed by a 14 d control period, so that each experiment lasted 7 weeks. Blood samples were taken on each of the last 3 d and faeces collected (using carmine markers) for the last 6 d of each diet period. 3. Plasma cholesterol, triglycerides and serum calcium concentrations were not altered by the addition of wheat fibre. 4. Wheat fibre caused a significant increase in faecal bulk and frequency. Faecal steroid excretion was the same in the last week of the first control and the high-fibre periods but increased in the subsequent control period.


Endogenous loss of leucine and methionine in adult male rats. 1. The fractional rate of loss of 14C and body-weight was measured in adult male rats after giving 14C-labelled methionine or leucine and maintaining rats for 30 d on a low-protein or a specific methionine+cystine-free diet: carcasses were then analysed for protein and fat 14C radioactivity. 2. The fractional loss of 14CO2 from [14C]methionine or [14C]leucine between day 20 and day 30 was always greater than the fractional loss of body-weight. 3. Carcass protein 14C radioactivity after giving [14C]leucine was higher than after giving [14C]methionine, but fat 14C radioactivity after either 14C-labelled amino acid was only a small proportion of the total body 14C radioactivity. 4. After correction of the fractional loss of 14CO2 for urinary 14C loss, but not body-weight loss, absolute amino acid loss was calculated using published values for methionine and leucine content of rats. 5. The best estimates of endogenous amino acid loss obtained using I-14C-labelled amino acids, expressed as mg/kg body-weight 0.75 per day were leucine 79, methionine 38.


The true absorption of cyst(e)ine from the ovine small intestine. Na234SO4 was infused into the rumen of a sheep fed a dried grass diet. 2. The digesta at the pylorus which contained 35S-labelled cyst(e)ine synthesized by the rumen microflora was collected and mixed with chromium EDTA. This material was infused into the duodenum of 35S free sheep. 3. Ileal digesta was subsequently collected and analysed for cyst(e)ine 35S and Cr. 4. The proportion of the labelled cyst(e)ine truly absorbed from the small intestine was 0-72 +/- 0-004.


Studies on gene control regions. 2. Enzymatic joining of chemically synthesized lactose operator deoxyribonucleic acid segments. The T4 polynucleotide ligase catalyzed joining of six chemically synthesized deoxypolynucleotides corresponding to lactose operator DNA has been investigated. Joining was studied using various combinations of segments. Joining reactions involving multiple sites and the formation of duplex operator DNA were complete in a few hours. Joining reactions involving a single site and the formation of only one strand of operator DNA required several days and repeated annealing in order to go to completion. These studies have permitted the synthesis on a preparative scale (several nanomoles) of operator duplexes and operator single strands.


Cross-complexing pattern of plant histones. Pea histones H2a, H2b, H3, and H4 have been isolated and their interactions studied by fluorescence anisotropy, light scatter, and circular dichroism. Histones H3 and H4 are almost identical in plants and animals, but plant histones H2a and H2b differ markedly from their mammalian counterparts. Pea H2b has a molecular weight approximately 20% greater than that of calf thymus H2b; the amino acid compositions of the two proteins are different. Calf thymus H2a exists as a single molecular weight species, while pea H2a exists as two species which differ by about 1500 daltons. The larger plant H2a is about 19% greater in molecular weight than calf thymus H2a. The smaller is about 8% greater. Despite these differences between calf and pea histones, the strong interactions between histone pairs H3 and H4, H2b and H4, and H2a and H2b, previously demonstrated for calf histones, also exist for pea histones. There are also weak interactions between pea H2a and H4 and between pea H2b and H3, and an interaction of intermediate strength between H2a and H3. The cross-complexing pattern of the plant histones is therefore the same as that reported for calf thymus histones [D'Anna, J. A., Jr., and Isenberg, I. (1974), Biochemistry 13, 4992], despite the dissimilarities of H2a and H2b.


A spectroscopic and kinetic investigation of anion binding to ascorbate oxidase. The binding of azide, fluoride, and cyanide to ascorbate oxidase has been investigated in detail. Both azide and fluorid inhibit the enzyme competitively with respect to ascorbic acid and noncompetitively with respect to oxygen. Cyanide inhibition is much more complex and also results in inactivation of the enzyme. The binding of azide and fluoride to the resting enzyme is partially competitive. Fluoride binds more strongly to the resting enzyme, while azide binds more strongly to the functioning enzyme. It is proposed that both azide and fluoride bind to type 2 copper and that this copper is also part of an ascorbate binding site. It seems likely that type 2 copper is a reductant binding site in all of the "blue" oxidases. This proposal is used to explain the effect of fluoride on the enzymes and also to suggest a mechanism for the internal electron transfer which is necessary for the reduction of oxygen to water.


Interaction of analogues of coenzyme A with choline acetyltransferase. The finding that methyl methanethiolsulfonate appears to inhibit choline acetyltransferase from squid ganglia not by reacting with a thiol group of the enzyme but by reacting with the thiol group of coenzyme A to form a competitive inhibitor of acetyl coenzyme A led to the synthesis of the ethyl, propyl, and 3-carboxy-4-nitrophenyl disulfides of CoA. The methyl disulfide of 1,N6-etheno-C0A, a fluorescent ligand, was also prepared. All the disulfides are powerful inhibitors of ChA, their Ki values being very similar. The Km values for acetylpropionyl-, and butyryl-CoA were also found to be similar; however, modification of the acyl group alter the Km values for choline. CoA, and dethia-CoA, showed similar abilities to be bound to ChA; however, the 3'-phospho groups of acetyl CoA and CoA appear to be of importance in interacting with the enzyme. 8-Anilino-1-naphthalenesulfonate is a competitive inhibitor of acetyl-CoA binding.


Studies of the subunit structure of wheat germ ribonucleic acid polymerase II. We have previously presented a rapid, high yield method for the large scale purification of homogeneous RNA polymerase II from wheat germ (Jendrisak, J.J., and Burgess, R.R.(1975), Biochemistry 14, 4639), and we now report a detailed study of its subunit structure. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate indicates that polypeptides with molecular weights of 220 000, 140 000, 40 000, 27 000, 25 000, 21 000, 20 000, 17 800, 17 000, 16 500, 16 000, and approximately 14 000 are associated with the enzyme. Two-dimensional polyacrylamide gel electrophoresis, by which the subunits were separated in the first dimension in the presence of 8 M urea at pH 8.7 and in the second dimension in the presence of 0.1% sodium dodecyl sulfate, indicates that the 40 000 molecular weight component is composed of two nearly identical polypeptides and that the low molecular weight components (smaller than or equal to 40 000) are acidic proteins except for the 25 000 molecular weight polypeptide.


Chemical synthesis of an octanucleotide complementary to a portion of the cohesive end of P2 DNA and studies on the stability of duplex formation with P2 DNA. A pyrimidine octanucleotide complementary to one of the cohesive ends of P2 DNA was chemically synthesized. Its sequence, d(C-T-T-T-C-C-C-C-OH), was verified by labeling it at the 5' end, followed by partial enzyme digestion and separation by a two-dimensional fingerprinting system. A single ribo-G residue was added to its 3' end using calf thymus deoxynucleotidyl terminal transferase. The resulting nonanucleotide primer was used in a detailed study on the stability of the duplexes formed in the partial as well as complete repair synthesis catalyzed by DNA polymerase I, at 5 degrees C in the presence of 70 mM potassium phosphate and 70 mM NaCl. The nonanucleotide primer was able to form a stable duplex with P2 DNA template only in the presence of DNA polymerase I. When the chain lengths of pyrimidine oligonucleotides were varied from 4 to 8 to test their abilities to serve as primers for the enzymatic repair synthesis, it was revealed that the minimum length required for the primer function is 8. Using the nonanucleotide as the primer and the right-hand cohesive end of the DNA as the template, repair synthesis was initiated simultaneously at the 3' end of the primer as well as at the right-hand 3' end of the DNA. This resulted in a decrease in the efficiency of repair synthesis at the 3' end of the primer, possibly due to the displacement of the primer by the enzyme. The enzyme was unable to displace the primer, when the primer was extended to a 13-mer prior to the initiation of repair synthesis at the 3'-OH end of the DNA. These data suggest that the strand displacement by DNA polymerase I at 5 degrees C in the presence of 70 mM potassium phosphate and 70 mM NaCl is not significant when the duplex is at least 13 nucleotides long. The efficiency of the repair synthesis at the 3'-OH end of the DNA-primer duplex could be increased by blocking the repair synthesis at the 3'-OH end of the DNA by converting it to 3'- phosphate. This method could be useful in DNA sequence analysis, where such specific repair synthesis is desired.


Energy transfer and the distribution of excitation energy in the photosynthetic apparatus of spinach chloroplasts. Equations are derived from our model of the photochemical apparatus of photosynthesis to show that the yield of energy transfer from Photosystem II to Photosystem I, phi T(II leads to I), can be obtained from measurements on an individual sample of chloroplasts frozen to -196 degrees C by comparing the sum of two specifically defined fluorescence excitation spectra with the absorption spectrum of the sample. Then, given that value of phiT(II leads to I), the fraction of the quanta absorbed by the photochemical apparatus which is distributed initially to Photosystem I, alpha, can be determined as a function of the wavelength of excitation from the same fluorescence excitation spectra. The results obtained in this study of individual samples of chloroplasts frozen to -196 degrees C in the absence of divalent cations, namely, that phi T(II leads to I)varies from a minimum value of 0.10 when the Photosystem II reaction centers are all open to a maximum value of 0.25 when the centers are all closed and that alpha has a value of about 0.30 which is almost independent of wavelength for wavelength shorter than 675 nm (alpha increases rapidly toward unity at wavelength longer than 675 nm), agrees quite well with results obtained previously from comparative measurements of chloroplasts frozen to -196 degrees C in the presence and absence of divalent cations.


The preparation of lipid-depleted bacteriorhodopsin. Bacteriorhodopsin, the protein of the purple membrane of Halobacterium halobium, was freed to the extent of 90-95% from the natural membrane lipids without loss of function. The residual lipid corresponded to less than 1 mol/mol of bacteriorhodopsin. Delipidation was achieved by treatment of the purple membrane with a mixture of the detergent dimethyldodecylamine oxide and sodium chloride. The detergent was removed by dialysis or by sucrose density gradient centrifugation. Analysis of the lipids removed and those still bound to bacteriorhodopsin was facilitated by the use of purple membrane preparations labelled with 35S, 32P, or 14C. The composition of the residual lipids associated with bacteriorhodopsin was similar to that of the total lipid in the purple membrane.


Positron lifetimes in phospholipid dispersions. Positron lifetimes have been determined in phospholipid dispersions. In fluid phosphatidylcholines, a lifetime of 3.3 ns is found, and a lifetime of 2.8 ns is found for frozen phosphatidylcholines. In dispersions where fluid and frozen phases coexist due to lateral phase separation, an intermediate lifetime is found.


A calorimetric examination of stable and fusing lipid bilayer vesicles. Mixed lipid samples containing dimyristoylglycerophosphocholine and small amounts of myristic acid were examined calorimetrically. Examination of multilamellar and small vesicle samples indicated that upon heating small vesicles combine to form more extended structures. An exothermic peak (at 19 . 5 degrees C) can be associated with the structural transformation. The enthalpy for this process, which may be interpreted as vesicle-vesicle fusion, is found to be approx.--2 kcal/mol.


The reactivity of human erythrocyte membrane cholesterol with a cholesterol oxidase. Cholesterol oxidase (EC 1.1.3.6, Brevibacterium sp.), which catalyzes the reaction: cholesterol + O2 leads to  delta4-cholestenone + H2O2, has no effect on the cholesterol of intact (human) erythrocytes and of "resealed" ghosts, when it is present only outside these ghosts. The cholesterol of "leaky" ghosts, of "resealed" ghosts with enzyme trapped within, and of "inside-out" vesicles, was completely oxidized. This pattern indicates that the inner ((cytoplasmic) membrane surface must be exposed to the enzyme for the reaction to occur, and that outer surface cholesterol only becomes reactive after the membrane  has been degraded by the oxidation of inner surface cholesterol. The enzymatic oxidations followed monotonic first-order kinetics, and hence gave no evidence to support the two states of cholesterol in the membrane that had been postulated earlier from studies on the plasma lipoprotein extraction of cholesterol from the membrane.


Topographical dissection of sheep erythrocyte membrane phospholipids by taurocholate and glycocholate. 1. Glycocholate and taurocholate removed significant amounts of membrane phospholipid from intact sheep erythrocytes before lysis of the cells occurred. The pre-lytic extract was enriched in sphingomyelin and correspondingly depleted in phosphatidylserine, phosphatidylinositol and phosphatidylethanolamine when compared to the original membrane. 2. In contrast, the phospholipid profiles of glycocholate and taurocholate extracts of unsealed ghosts, made at the same bile salt concentrations, were similar to that of the whole membrane. 3. These observations are related to the topography of the phospholipids in the membrane and to some aspects of bile formation.


On the specific association of porcine erythrocyte catalase caused by formation of disulfide cross-links. Porcine erythrocyte catalase (hydrogen-peroxide:hydrogen-peroxide oxidoreductase, EC 1.11.1.6) has been purified from porcine blood by DEAE-cellulose column chromatography, ammonium sulfate fractionation and CM-cellulose column chromatography. The purified enzyme was found to associate into larger molecules than the native one when it was stored at 4degrees C for more than one week. The associated molecules can be detected by gel filtration on a Bio-gel A-1.5 m column and disc gel electrophoresis as well as ultracentrifugal analysis. Molecular weights of the associated catalase molecules were about 500 000, 750 000, 1 000 000 and so forth estimated by gel filtration and disc gel electrophoresis, corresponding to dimer, trimer and tetramer respectively, of a native molecule (monomer) with a molecular weight of about 250 000. The association of catalase molecules is found to be time-dependent and to proceed seemingly from monomer through dimer as an intermediate. From the effects of several thiol reagents or reducing reagents on the association process and spectrophotometric titration of SH groups, it is inferred that this specific association of procine erythrocyte catalase is caused by formation of intermolecular disulfide cross-links due to air oxidation of SH groups in the protein moiety.


Purification and molecular properties of bovine heart pyruvate kinase. A rapid method is presented for the purification of pyruvate kinase (ATP : pyruvate 2-O-phosphotransferase, EC 2.7.1.40) from bovine heart. The enzyme obtained is homogeneous by criteria of sodium dodecyl sulphate polyacrylamide electrophoresis and ultracentrifugation and has a specific activity of 260 units/mg. It is a tetramer of 220 000 daltons and S020,w = 10.0 S and possesses no free amino-terminal residue. The amino acid composition is similar to that of the M1 isozyme of rabbit and bovine skeletal muscle. The enzyme is subject to polymerisation to a hexamer of the basic tetramer. The polymeric species has a molecular weight of 1320 000, is promoted at low ionic strength and is undetectable at ionic strength greater than 0.2 by either sedimentation equilibrium or sedimentation velocity measurements. The polymerisation is independent of temperature in the range 5--20degrees C implying that charge interactions rather than apolar interactions are responsible for the process.


Carboxylesterases of human brain extract. Purification and properties of a butyrylesterase. 1. A carboxylesterase (carboxylic-ester hydrolase, EC 3.1.1.1) from human brain extract was prepared to purity using DEAE-cellulose, Sephadex G-200, and fractionation with (NH4)2SO4. The yield was about 20%. 2. Esters of butyric acid were split faster than esters of acetic, propionic and valeric acid, and the enzyme is tentatively designated as a butyrylesterase. Thiocholine esters were split at low rates. 3. The molecular weight was estimated as 340 000 using gel chromatography on Sephadex G-200. In isoelectric focussing the enzyme was resolved into several peaks (pI 4.0--4.7). The low isoelectric point does not seem to be due to terminal sialic acid residues. 4. The enzyme was irreversibly inhibited by diethyl-p-nitrophenyl phosphate (ki = 206 mol-1 - 1 - s-1) and by diisopropylfluorophosphate. The carboxylesterase inhibitor bis-p-nitrophenyl phosphate and eserine did not inhibit the enzyme. 5. The enzyme was progressively inhibited by p-hydroxy-mercuribenzoate, and reactivated by dithiothreitol and 2-mercaptoethanol. N-Ethylmaleimide inactivated the enzyme very slowly, whereas iodoacetate and iodoacetamide were without effect. 6. Ca2+, Mg2+, and Zn2+ or EDTA did not influence the enzyme activity.


Purification and properties of human liver peptidase. The peptidase from human liver was purified using L-Leu-L-Leu as a substrate, in adapted purification techniques including treatment with n-butanol, acetone precipitation, ammonium sulfate fractionation, DEAE-cellulose chromatography, Sephadex G-150 gel filtration and CM-cellulose chromatography. The purified enzyme exhibited homogeneity in disc electrophoresis. The molecular weight of the enzyme was estimated to be 130 000 by Sephadex G-200 gel filtration. The isoelectric point of the enzyme was found to be pH 5.6. The enzyme was activated by Mn2+ and inhibited by o-phenanthroline. L-Leu-L-Leu and L-Phe were hydrolyzed effectively by the peptidase. By electrophoresis on Cellogel, the electrophoretic mobility of purified enzyme was same as that of the peptidase in serum of patients with hepatic disease.


Studies on the enzymatic hydrolysis of polyglutamyl folates by chicken liver folyl poly-gamma-glutamyl carboxypeptidase. II. Structural studies. Further studies on the purified chicken hepatic folyl poly-gamma-glutamyl carboxypeptidase (peptidyl-L-glutamate hydrolase, EC 3.4.12.10) have elucidated some of the structural characteristics of the enzyme. Various analytical studies described reveal 424 amino acid residues in the isolated native enzyme with molecular weight of around 57 900. beta-Mercaptoethanol (14.3 mM) activated the enzyme 2.2-fold and induced reductive cleavage of an interchain disulfide linkage resulting in the splitting of the native enzyme into two active polypeptides (molecular weights 43 000 and 18 000). The constituent polypeptides have identical NH2-terminal residues (valine) and exhibit a high degree of sequence homology as revealed by finger print analyses of their tryptic digests. The 10-fold greater sensitivity of the reductively cleaved enzyme to p-chloromercuribenzoate would imply that active site related sulfhydryl groups are not readily accessible in the native enzyme. Ionic strength effects in the presence of Mn2+ and Na+ and the presence of low urea concentration (0.55 M) result in a further up to 5-fold stimulation of reductively cleaved native enzyme. Citrate inhibited and phosphate induced autolytic degradation of the enzyme. The physiological role of gamma-glutamyl carboxypeptidase has been discussed.


[Study fo sterols from brown seaweeds of the genus Cytoseira. Identification by gas-liquid chromatography coupled with mass spectrometry (author's transl)]. All the samples of brown seaweeds (Cystoseria) that we have studied present the same deltas sterols fucosterol, 22 trans-dehydrocholesterol, brassicasterol, 24-methylene cholesterol as well as cystosterol, a new C27 sterol. This sterol has been submitted to gas-liquid chromatographic-mass spectrometric analysis.


Stereoconfiguration of bisphosphatidic and semilysobisphosphatidic acids from cultured hamster fibroblasts (BHK cells). Monolayers of hamster fibroblasts (BHK cells) were incubated in Eagle's minimal essential medium under conditions where an increase in the levels of all cellular bisphosphatidic acids takes place. Bisphosphatidic acid and semilysobisphosphatidic were isolated from these cells and subjected to strong alkaline hydrolysis. Stereochemical analysis of the hydrolysis products revealed that the majority of the molecules of both lipids are derivatives of sn-1-glycerophospho-sn-1'-glycerol, the structure previously found to be the "backbone" of lysobisphosphatidic acid, (bis(monoacylglycerol)phosphate) from BHK cells and other sources. This finding suggests a close metabolic relationship between the three bisphosphatidic acid derivatives of BHK cells.


Squalene epoxidase and oxidosqualene lanosterol-cyclase activities in cholesterogenic and non-cholesterogenic tissues. Squalene epoxidase and oxidosqualene lanosterol-cyclase activities have been studied in normal mammalian cholesterogenic and non-cholesterogenic tissues. This paper describes the kinetic conditions of measurement of these two enzymatic acitivities and their results. Oxidosqualene lanosterol-cyclase is a widespread enzyme, present in all cholesterogenic and non-cholesterogenic tissues. However, the level of squalene epoxidase is very low in non-cholesterogenic tissues. However, the level of squalene epoxidase is very low in non-cholesterogenic tissues. The effects of subcellular fractionizing and of the physico-chemical state of squalene incubated in vitro on squalene epoxidase activity are discussed.


Long-chain diglycerol tetraethers from Thermoplasma acidophilum. The C40 isopranol-containing glycerol ether residues which characterize the complex lipids of the extreme thermoacidophile Thermoplasma acidophilum were isolated and purified from the glycolipid and phospholipid fractions. The glycerol ether, as well as the acetate and methoxy derivatives were characterized by thin-layer, gel-permeation and gas-liquid chromatography, infrared, nuclear magnetic resonance and mass spectrometry and by vapor phase osmometry. The glycerol ethers are proposed to be unique fully saturated diglycerol tetraethers, primarily C86H172O6, Mr 1300, which contain two sn-2,3-glycerol residues bridged through ether linkages by two C40 isopranoid branched diols.


The polar lipids of group B Streptococci. I. Glucosylated diphosphatidylglycerol, a novel glycopholipid. 1. From group B Streptococci a novel glycophospholipid was isolated, which contained D-glucose, glycerol, acyl groups and phosphorus in a molar ratio of approx. 1:3:4:2. It was established to be 2'-O-alpha-D-glucopyranosyl-, 1',3'-bis-(1,2-diacyl-sn-glycero-3-phospho)-glycerol. 2. The structure of the deacylated core was accomplished by analyses of the breakdown products obtained on (i) strong alkaline hydrolysis, (ii) Smith-degradation, and (iii) periodate oxidation with subsequent hydrazinolysis. The four acyl groups were located by sequential degradation of the native lipid with phospholipase A2 and 98% acetic acid. 2. In group B Streptococci approximately 25% of diphosphatidylglycerol occurs in the form of its glucosylated derivative which accounts for 18% of the lipid phosphorus. The glucosylated phosphatidylglycerol analogue could not be detected. A phosphoglucolipid, however, was present, which was tentatively identified as glycerophosphodiglucosyldiacylglycerol.


Snake venom toxins. The amino acid sequence of toxin Vi2, a homologue of pancreatic trypsin inhibitor, from Dendroaspis polylepis polylepis (black mamba) venom. The amino acid sequence of venom component Vi2, a protein of low toxicity from Dendroaspis polylepis polylepis venom was determined by automatic sequence analysis in combination with sequence studies on tryptic peptides. This protein, the most retarded fraction of this venom on a cation-exchange resin, is a homologue of bovine pancreatic trypsin inhibitor consisting of a single chain of 57 amino acid residues containing six half-cystine residues. The active site lysyl residue of bovine trypsin inhibitor is conserved in Vi2 although large differences are found in the rest of the molecule.


Isolation of phosphorylated acid chloroform/methanol-soluble proteins from live frog muscle. About 6-7% of the total proteins from trichloroacetic acid-washed and freeze-dried frog muscle could be extracted with acid chloroform/methanol. Three of these proteins were found to be phosphorylated in  the live frog. They were purified to apparent homogeneity by gel chromatography and preparative gel electrophoresis. The apparent molecular weights, determined by sodium dodecyl sulfate gel electrophoresis, were 34 000, 19 000 and 10 000. Each phosphorylated protein contained 3 mol of a covalently bound neutral sugar but they did not contain any tightly bound lipids. All three proteins incorporated 32P into serine phosphate. The 10 000 dalton protein, which had the highest specific radioactivity contained an unusually high proportion of serine, 14% of the total amino acids. It also did not stain with Coomassie Blue.


ATP binding to human hemoglobin in the presence and absence of magnesium ions investigated with 31P NMR spectroscopy and ultrafiltration. The addition of 3 mM hemoglobin to 1-10 mM ATP solutions at pH 6.75 resulted in a linear relationship between the change in chemical shift (deltadelta) of the gamma-phosphate of ATP and the percent ATP bound to hemoglobin. The data points obtained with oxyhemoglobin and deoxyhemoglobin fell on the same straight line. In the presence of 3 mM Mg2+, the delta delta decreased curvilinearly as the percent ATP bound was raised. In this case, the percent ATP bound to deoxyhemoglobin was greater than to oxyhemoglobin at the same delta delta value, indicating that the extra ATP binding occurs through groups other than phosphate.


A hydrophobic quencher of protein fluorescence: 2,2,2-trichloroethanol. Previously the neutral fluorescence quenching probe, acrylamide, was employed to determine the degree of exposure of tryptophan residues in  proteins. A less polar neutral quencher 2,2,2-trichloroethanol (trichloroethanol) was used in the present work to investigate whether it would preferentially interact with apolar regions of proteins. For most proteins studied, the degree of quenching by trichloroethanol is found to be about the same as with acrylamide. However, for human and bovine serum albumin hydrophobic interactions between trichloroethanol and these proteins occur, leading to an exalted quenching. The fluorescence quencher thus senses the presence of a hydrophobic domain in the vicinity of the tryptophan residues in these proteins. Trichloroethanol is shown to induce conformational changes in certain proteins and to be a potentially useful quencher for proteins having predominantly tyrosine emission.


The properties of Corbicula sandai apoferritin. Corbicula sandai apoferritin possesses physical properties different from apoferritins of other species. The native molecular weight was estimated from its s020,w of 18.7 S to be about 503 000. Empirical molecular weight estimation methods in denaturing solvents yielded a molecular weight estimate for the constituent polypeptide chain of 23 000. The circular dichroic spectrum of C. sandai apoferritin was significantly different from other apoferritins and it was immunologically unreactive with rabbit anti-human ferritin antisera.


Preparation of monoiodotyrosine-13-glucagon. Glucagon was iodinated with the lactoperoxidase method at pH 10.0 in the presence of propylene glycol using a substitution of 0.3 g-atom I/mol glucagon. Under these conditions the reactivity of the iodine to tyrosine at position 13 is found to be 4-fold that of the tyrosine at position 10. The amount of diiodotyrosine was less than one-twentieth that of the monoiodotyrosine at either tyrosine residue. Relatively pure monoiodo[125I]tyrosine-13-glucagon can be separated from other iodoglucagons by means of DEAE-chromatography. Such a homogeneous preparation with a known position of the iodine makes it possible to study a specific interaction between the monoiodoglucagon and the glucagon antisera or the glucagon receptor.


Application of affinity electrophoresis to the study of antigen-immunoadsorbent association equilibrium. Affinity electrophoresis is used to study the interaction of complementary biological molecules. Using Scatchard plots, this method gives a partition ratio between immunoadsorbent and soluble antigen or antibody. In the present work, affinity electrophoresis, used to study the interaction between human serum albumin and its insolubilized antibodies, allows a measure of the system's affinity under specific conditions, in particular, the electric field.


A carbon monoxide derivative of ruthenium (II) myoglobin probe of heme protein conformation. A synthetic carbon monoxide complex of ruthenium (II) myoglobin has been reconstituted from apomyoglobin and the carbon monoxide of ruthenium (II) mesoporphyrin IX. This synthetic myoglobin complex shows an absorption spectrum with normal Soret and beta bands, and a split alpha band. The alpha-band splitting is not observed in the spectrum of the carbon monoxide derivative of ruthenium (II) mesoporphyrin IX in pyridine, even though the width of the alpha band in pyridine is narrower than in theprotein. The separation between two alpha bands in the spectrum of the protein is reduced from 7.5 to 6 nm in the presence of 2 M NaCl. This observation is interpreted in terms of perturbation of concentrated NaCl on the protein. The separation between the maxima of two alpha bands in the spectrum of the ruthenium (II) myoglobin also becomes smaller with decrease in pH from 7 to 4.5, and this process involves the distal histidine. The alpha band splitting in this protein complex is interpreted in terms of rhombic distortion of the square planar symmetry of the metal porphyrin in the protein.


Protein sequencing by computer graphics. A computer graphics system has been used to fit a putative sequence to an electron density map of a sea snake neurotoxic protein at 2.2 A resolution. The complete sequence of this small protein could be determined from the map with very little ambiguity. In two places probable errors in the published chemical sequence were detected. This is the first instance in which a complete three-dimensional structure was solved with the use of computer graphics alone, without the construction of a physical model.


Specific refractoriness of adenylate cyclase in skin to epinephrine, prostaglandin E, histamine and AMP. The cyclic AMP level  in pig skin (epidermis) increases markedly after incubation with epinephrine, prostaglandin E, histamine or adenosine 5'-monophosphate. This increase is transient and "spiking" is the consistent response to these four stimulators. The "spiking" is due to a non-responsiveness or refractoriness which develops within minutes and is specific to any one stimulating hormone but not to the others. The addition of inhibitors of protein syntheses did not prevent the development of the refractoriness. Adenylate cyclase and phosphodiesterase activities measured in skin homogenates prepared from skin samples taken before, during and after the "spiking" did not change significantly. The hormone-induced refractoriness in this skin system appears to be due to a specific, localized loss of function of the adenylate cyclase system.


The effect of serum protein fractions on liposome-cell interactions in cultured cells and the perfused rat liver. We have studied the interactions of liposomes with human skin fibroblasts and mouse P815Y mastocytoma cells in culture, and the perfused rat liver, with the following findings. 1. In all the systems studied serum was found to cause an increase in the uptake of a [14C]cholesterol label into cells from anionic and neutral liposomes and a decrease in the uptake of the label from cationic liposomes. 2. Evidence suggests that albumin enhances the exchange/transfer of [14C]-cholesterol between liposomes and cultured cells. 3. With [14C]cholesterol in the liposome bilayer and [3H]methotrexate entrapped in the aqueous spaces of the liposome, the alpha- and beta-globulin fractions of serum decreased the transfer of both labels from cationic liposomes into cultured cells and the perfused rat liver. The beta-globulin fraction caused increased leakage of methotrexate from fluid liposomes of all charges. 4. The alpha- and beta-globulin fractions of serum appear to enhance the uptake of anionic liposomes into the perfused rat liver.


On arabinose as a component of brain hyaluronate. Confirmation by chromatographic, enzymatic and chemical ionization-mass spectrometric analyses. The controversy about the presence of the pentose arabinose in brain hyaluronate was reinvestigated using modern analytical tehcnics. The purified bovine brain hyaluronate contained the neutral sugars: arabinose, 0.18%; glucose, 0.05%; and fucose, 0.22%. The confirmation of the presence of arabinose was obtained by paper and thin layer chromatography of the neutral sugars in deionized hyaluronate hydrolysate. Gas-liquid chromatography of the aldononitrile peracetate of the pentose isolated by preparative paper chromatography gave a single distinct peak, corresponding to standard arabinose on three columns packed with three different phases. Chemical ionization data and mass spectrum of the aldononitrile peracetate derivative agreed with those of the authentic arabinonitrile tetracetate. Analysis of the isolated pentose with the help of the enzymes L-arabinose isomerase and L-ribulose kinase, which are specific for their substrates, further established its identity as L-arabinose.


Immobilized ACP and electrofocusing used to purify the palmityl-CoA-ACP-transacylase from Mycobacterium smegmatis. This paper describes the first purification to electrophoretic and chromatographic homogeneity of a palmityl-CoA-ACP-transacylase. Palmityl-CoA-ACP-transacylase of Mycobacterium smegmatis shows two interconvertible forms in monomer-oligomer correlation. Taking advantage of this interconvertibility properties a 800-fold purification of the enzyme was recently achieved (Kervabon et al (1976) Biochimie, 58, 647-656). The present report describes the use of immobilized ACP and electrofocusing to obtain a 880-fold and a 5 500-fold purification respectively. The enzyme thus purified, pI 4.7, showed a single band upon polyacrylamide gel electrophoresis. Chromatography on Sephadex G-150 revealed only the peak of the monomer. When the enzyme solution containing 200 microng/ml of protein was concentrated by vacuum dialysis to approximatively 1 mg/ml, gel electrophoresis showed two bands one corresponding to the monomer and the other to the oligomer formed.


Evidence that proteins bound to the polysomal messenger RNA exist also free in the cytoplasm of HeLa cells. The cytoplasm of HeLa cells contains a series of proteins with affinity for RNA. These proteins were detected by affinity chromatography on RNA-Sepharose. Several proteins are tightly bound to RNA; there is no preference for mRNA as the same proteins bind to any free RNA, even poly(A). At least two of these proteins coincide with proteins of genuine ps-mRNP-proteins (Mr. 50 000 and 75 000). The column yields a protein (Mr. 37 000) which has an affinity for the matrix rather than for the coupled RNA. Complexes formed from mRNA and cytosol proteins do not bind to ribosomes, whereas ps-mRNPs does. We suggest that some of the polysomal mRNP proteins pre-exist unbound in the cytosol.


[Effects of glucocorticosteroids on enzymatic activity in the urea cycle in fetal rat liver]. The five urea cycle enzyme activities of rat liver are followed during late fetal period and the first day of life. All five enzymes exhibit relatively low activities in foetal liver and a rapid postnatal increase. Lack of glucocorticosteroid (after hypophysectomy in utero) induces an important decrease of activity of three enzymes: carbamyl phosphate synthetase, ornithine transcarbamylase and argininosuccinate synthetase. Treatment with hydrocortisone acetate on decapitated fetuses results in a marked stimulation of the activity of four of the enzymes: carbamyl phosphate synthetase, ornithine transcarbamylase, argininosuccinate synthetase and arginase. Premature induction of carbamyl phosphate synthetase activity is obtained after intraperitoneal injection with hydrocortisone acetate at 16,5 days of gestation.


An iron-containing superoxide dismutase from the strict anaerobe Desulfovibrio desulfuricans (Norway 4). Superoxide dismutase, the enzyme which catalyzes the dismutation of superoxide free radicals (formula: see text) has been purified to homogeneity from the strict anaerobe sulfate-reducing bacterium Desulfovibrio desulfuricans (Norway 4). Its molecular weight is 43,000 and it is composed of two subunits of equal size which are not covalently bound. The enzyme was found to contain iron by atomic absorption and the absence of acid-labile sulfur indicates that it is not an iron-sulfur protein. Electron paramagnetic resonance spectrum revealed that iron occurs in a high spin ferric form. The ultraviolet and visible absorption spectra of the enzyme are presented, as are the results of amino-acid analysis. The data reported allow to conclude that this superoxide dismutase isolated from a strict anaerobe exhibits similar physico-chemical properties as compared to the iron-containing dismutases found in aerobic microorganisms. The significance of the presence of a superoxide dismutase in this strict anaerobe sulfate reducer is discussed.


[Mechanism of histone action on mitochondrial energetics. Proton and cation transport across membrane]. The effect of lysine-rich histone on mitochondria was studied with respect to the cation transport and proton uptake across the membrane. Histone does not affect Ca2+ uptake or level in mitochondria. It extrudes Mg2+ by 30% with the utilization of substrates; in the presence of rotenone extrusion drops to about 18%. Dinitrophenol and ADP prevent the effect of histone. Mg2+ uptake is inhibited by 12% by histone. K+ extrusion by histone (by 50%) which depends on the oxidation of substrates, is enhanced by phosphate, and Mg2+ decreases (by 50%) the K+ extrusion. Histone prevents to some extent extrusion of K+ by Ca2+. Histone does not affect Na+ level in mitochondria, it blocks (by 50%) Na+ uptake in the presence of gramicidin, probably by forming a gramicidin-histone complex. Histone blocks proton uptake by mitochondria incubated in the presence of valinomycin or DNP. In the presence of DNP valinomycin-induced H+ uptake is not affected by histone. H+ uptake induced by nigericin is not affected by histone as well.


[The state of phospholipase D in solution and its catalytic activity]. Functioning of water-soluble phospholipase D from cotton seeds is studied on two phases contact area (liquid-liquid, liquid-solid substance) and on the surface of mixed lecitine and sodium dodecylsulphate micelles. It is found that water-soluble phospholipase D, which normally has no catalytic activity, is capable to hydrolyse its substrates in the presence of organic solvents, solid adsorbents and sodium dodecylsulphate. The data obtained show that in all the cases studied the activation observed is due to adsorption immobilization of the enzyme. K lambda and K alpha constants are introduced, which are characteristics of immobilyzing ability of agents-matrices for immobilization. Phase transitions, which take place in heterogenous system (enzyme-activator-substrate-water solution), are found to be a necessary condition for the enzyme activation. A hypothesis, that catalytical activity of water-soluble phospholipase D is inherent of the adsorbed enzyme, is discussed on the basis of the data on comparative study of adsorbed and water-soluble enzymes.


[Inhibition of nonenzymatic ATP hydrolysis by its binding with the cationic detergent micelles]. The rate of ATP hydrolysis is shown to decrease sharply by its binding with mycelles formed from cetyltrimethyl ammonium bromide (CTAB) (pH 9.0, 20 degrees C). Under these conditions only trace amounts of hydrolytic products have been found in the reaction mixture after a two-months standing. In the absence of mycelles the ATP hydrolysis is practically completed in 7 days. The energies of ATP, ADP and Pi binding with mycelles and at the interface of the "octane - water" system with previously adsorbed CTAB have been measured. The estimates demonstrate that the binding of one negative charge of phosphate anion or that of nucleotides with CTAB at the interface of the "octane - water" system corresponds to the change in free energy, i.e. -3.5 ccal/mol. The sharp deceleration of ATP hydrolysis cannot be due to the shift in the equilibrium of ATP hydrolysis reaction towards the formation of the pyrophosphate bond under conditions when all reaction components are bound to the surface of the mycelles.


[Study of nystatin effect on nuclear proteases of dog kidney causing degradation of acid-soluble proteins of chromatin]. A relationship between the previously observed degradation of dog kidney chromatin histones induced by polyene antibiotics, and a change in enzymatic activity of neutral histone protease or proteases localized on the outer nuclear membrane was studied. It was shown that in the course of proteolysis of chromatin isolated from the nuclei after treatment by Triton X-100 nistatin inhibits neutral histone proteases as can be evidenced from the amount of the amino groups formed and the data of electrophoretic analysis of chromatin histones. The inhibitory effect of nistatin is sharply enhanced with an increase in its concentration. It is found that in the course of proteolysis of chromatin isolated from the nuclei untreated with Triton X-100, nistatin increases the degree of acid-soluble protein degradation. The latter effect is probably due to the stimulation of the enzymatic activity of proteases localized on the outer nuclear membrane by nistatin.


[Interaction of aspartate aminotransferase with aminooxyalkylcelluloses]. Interaction of aspartate aminotransferase with various aminooxycelluloses capable of reacting with carbonyl compounds to form oximes has been studied with respect to the distance of the H2NO-groups from the polymer matrix. Aminotransferase does not react with aminooxycelluloses, when the amono-oxygroups are located at small distances from the matrix. When these celluloses interact with the enzyme in the presence of the substrate amino acid, we manage to obtain the amino-form of amino-transferase containing no pyridoxylidine form quantitatively. The cellulose with the H2NO-groups located at considerable distances from the polysaccharide matrix, form the "aminotransferase oxime--aminooxycellulose" complex, which is subsequently split either to a choloenzyme or apoenzyme of PLP and aminooxycellulose, depending on the experimental conditions used. The adsorbents under study may present some interest both in terms of isolation and purfication of pyridoxal enzymes and their effects on enzymatic systems, which contain carbonylic compounds.


[Activation of latent RNAse activity of RNP particles from rat skeletal muscles]. The dependence of activation of latent RNAse of RNP particles isolated from rat skeletal muscles, on the concentration of K+ and Mg2+ in the incubation medium and on temperature was studied. During a short-term exposure (20 min 18 degrees) of RNP particles in the buffers containing K+ at concentrations varying from 0.05 M to 0.25 M and Mg2+ at concentrations from 0.001 M to 0.01 M no effect of endogenous ribonuclease was observed. It was shown that a significant activation of latent RNAse occurs during the incubation at room temperature in 24 hours provided that the incubation medium contains Mg2+ at concentrations not higher that 0.004 M. In the presence of 0.004 M Mg2+ degradation of polysomal mRNA and partial degradation of 18 S--rRNA takes place. At Mg2+ concentration as low as 0.001 M not only mRNA but also both rRNAs are accessible to the action of activated ribonuclease. 20 min heating of RNP particles up to 55 degrees C causes insignificant degradation of the polysomes and 18 S--rRNA. The increase in temperature by 5 degrees c results in the activation of latent RNAse followed by an almost complete degradation of mRNA and rRNA. The relationship between the integrity of the ribosomal structure and activation of latent ribonuclease is discussed.


Erythrocyte catechol-o-methyltransferase activity in primary affective disorder. Previous studies have suggested that the activity of erythrocyte catechol-O-methyltransferase (COMT) may be reduced in women with bipolar and particularly unipolar affective illness. More recently, increased COMT activity in both men and women with affective disorder was reported. The activity of COMT in erythrocytes was determined in 184 outpatients with primary affective disorder at the Lithium Clinic of the New York State Psychiatric Institute. COMT activity was determined by a modification of the Axelrod and Cohn method with dopamine as substrate. This change resulted in an apparent threefold increase in the values for COMT activity; however, when both methods were compared, the results correlated (r = 0.97, p less than 0.001, N = 37). We found that the values for women were not significantly lower than those for men. In addition, there was no difference between patients with affective disorder and controls. Further, no difference was demonstrated between patients diagnosed as bipolar or uni-polar. Parameters such as mood, medication, and inpatient or outpatient status had no effect on COMT activity. The results of previous studies are discussed in an attempt to reconcile the different results.


Coexisting tardive dyskinesia and parkinsonism: a case report. A case report is presented of a patient with remitting tardive dyskinesia coexistent with neuroleptic-induced parkinsonism. The patient received benztropine and physostigmine challenges on successive days and the effects on both syndromes are described. The clinical course of both diseases is discussed in terms of a balance hypothesis between dopaminergic and cholinergic neurotransmission.


Reduction of progesterone release rate through silicone membranes by plasma polymerization. An investigation was performed to determine the effectiveness of thin plasma-polymerized films and CASING for controlling the flux of progesterone through poly-(dimethyl siloxane) membranes. It was concluded that up to 40-fold reductions in flux could be achieved with plasma-polymerized films. The principal factors affecting the extent of flux reduction were the nature of the monomer used (ethylene, ethane and tetrafluoroethylene), the film thickness, and the deposition conditions. CASING (Crosslinking by Activated Species of Inert Gases) of the poly-(dimethyl siloxane) membrane surface also proved effective in reducing the flux of progesterone through it.


Shear-induced hemolysis with commercial and glow discharge silicones. The hemolytic characteristics of 14 different polydimethyl-siloxane materials were studied, using a rotating disk device to shear whole human blood for 6000 sec. Disk surfaces included commercial Silastic products with and without silica filler, coatings formed from hexa-methyl and tetra-methyl disiloxane monomer vapors by glow discharge, and heparinized silicones. Results are presented in the form of kinetic hemolysis curves (KHC)--plots of plasma hemoglobin vs. time--and analyzed in terms of the slopes of these curves in the 1500-6000 sec region where the KHC is nearly linear. It was found that most silicone products are low-hemolysis materials, comparable to polyethylene (PE) but often about 20% less ehmolytic; one product was superior by about 40%. The only consistent exceptions to this were the glow discharge surfaces from HEXA-methyl disiloxane (15-80% poorer than PE) and heparinized surfaces (70-280% more hemolytic than PE).


Evidence for conformeric states of rhodopsin. Spectrophotometric measurements of metarhodopsin II appearance are made on five different kinds of rhodopsin preparations. Although the preparations differ greatly in their rhodopsin: phospholipid ratio, the meta II kinetics in all of them are strikingly similar in certain respects. Meta II appearance kinetics in all of the preparations are best described by two and only two exponentials. The ratio of these two rates is always about 5. The fast fraction: slow fraction ratio depends upon temperature. These fractions are reversibly convertible in the dark, and are interconverted on a time-scale which is long compared to the meta II appearance rate. It is shown that the kinetics of the earlier step in the bleaching sequence, viz., lumi-leads to meta I, is also described by double exponentials. Again the ratio of rates is ca. 5 and the fast-slow fractions correspond to those found in the meta I leads to meta II step. It is proposed that these facts support an hypothesis for the existence of two conformeric states of rhodopsin which are in thermal equilibrium. Thermodynamic parameters associated with this proposed equilibrium are presented.


A proposed model for rhodopsin in photoreceptor membranes. Transient electric birefringence studies have been made on bovine rhodopsin solubilized in the detergent lauryldimethylamine oxide from glutaraldehyde fixed rod outer segment (ROS) membranes. It was found that fixation caused no appreciable differences in the measured relaxation times when compared with unfixed ROS. On the basis of these findings a model for the orientation of rhodopsin in photoreceptor membranes is proposed which accounts for translational diffusion and two modes of rotational diffusion. The proposed model is related to a number of experimentally determined biophysical properties reported in the literature.


Photoreactions of bacteriorhodopsin.

Photoreactions of bacteriorhodopsin. Bacteriorhodopsin is a membrane-bound light energy transducer which generates an electrochemical proton gradient. It undergoes a cyclic photoreaction in which five intermediates have been identified. During the cycle it releases a proton from one surface of the membrane and takes up a proton on the opposite surface. The active chromophore consists of retinal bound through a Schiff base to the protein. The Schiff base is deprotonized during the photoreaction cycle and appears to be involved in the transport of protons through the membrane. The retinal may also undergo an isomerization.