Cultivation of erythrocytic stages.

Cultivation of erythrocytic stages. Two methods are described for the continuous cultivation of Plasmodium falciparum. One method provides for a continuous slow flow of medium over a settled layer of blood cells and does not require daily attention. With the other method, carried out in Petri dishes, the medium has to be changed manually every day. The problems and possibilities of continuous cultivation are discussed.


Population studies of Plasmodium vivax. 1. The theory of polymorphism of sporozoites and epidemiological phenomena of tertian malaria. The understanding of clinico-epidemiological phenomena of tertian malaria has been the subject of controversy. The authors suggest a system of postulates which give a non-contradictory explanation of the phenomena of relapses and long incubation. The main idea is that the duration of exoerythrocytic development of Plasmodium vivax is a polymorphic characteristic controlled by a set of genes. According to these postulates sporozoites may be subdivided into two groups designated as tachysporozoites and bradysporozoites, responsible for early and late manifestations, respectively. The logical analysis of the system suggests that it does not contradict the experimental facts. Moreover, the theory of polymorphic sporozoites permits an explanation and quantification of interrelations between different phenomena. The authors stress that the variation of genes is much greater in natural populations of parasites than in individual isolates and strains and therefore that the features of strains do not fully reflect the features of populations. Classical laboratory experiments should be combined with epidemiological experiments which allow a study of the population as a whole. The methodology of experiments to be undertaken in further investigations of the long latency period is discussed.


Population studies of Plasmodium vivax. 2. Distribution of manifestations in foci of tertian malaria. The authors investigate a mathematical model based on the theory they proposed in a previous publication. The model fits field data collected in re-established foci of tertian malaria. The patterns of distribution of manifestations of tertian malaria among the population may readily be explained on the basis of the theory of polymorphism of sporozoites.


Experimental animals and in vitro systems in the study of lymphocytic choriomeningitis virus. The history of lymphocytic choriomeningitis (LCM) research is reviewed from the point of view of whether the main discoveries concerning LCM pathogenesis have stemmed from animal or in vitro research methods. Most of the results initially stemmed from animal experiments, but in recent years recourse has increasingly been made to in vitro techniques to confirm and amplify the animal-based conclusions.Different research approaches are discussed and an attempt is made to assess the role of in vitro versus animal methods in obtaining knowledge on this virus. The same analysis is applied to the future needs and trends in arenavirus research.


Controlled field trial of a high-dose oral killed typhoid vaccine in India. A controlled field trial of a modified vaccine containing Salmonella typhi, TY 2 strain, in a dose of 400 x 10(9) killed organisms per capsule, was carried out in schoolchildren aged 6-17 years. The children were followed up for 6-8 months. There were 66 cases of typhoid fever in the vaccine group (7312 children) and 54 in the control group (7292 children who had received a placebo). This difference is not statistically significant. The results confirm that, as found in earlier trials, oral typhoid vaccine, in the dosages used so far, gives no protection.


Pathophysiological differences between paired and communal breeding of male and female Sprague-Dawley rats. Sexually mature, male and female Sprague-Dawley rats were housed in large communal breeding cages or in smaller paired breeding cages. Virgin control rats of the same age were housed similarly but segregated by sex. Breeders became obese, developed a fatty liver, and showed elevated levels of triglycerides, free fatty acids, and cholesterol. Breeders had high blood pressure, enlarged hearts, hyperglycemia, and islet beta cell degranulation. Serum enzymes, creatine phosphokinase, serum glutamic oxalo-pyruvic transaminase, serum glutamic pyruvic transaminase, lactate dehydrogenase, and blood urea nitrogen levels were elevated in breeder rats. The adrenal glands of male breeders appeared hyperactive; the adrenal glands of female breeders were thrombosed and appeared to be hypoactive. Male breeder rats developed microscopic aortic lesions only; female breeders developed advanced calcific aortic sclerosis. Male breeders kept in active stud service manifested the most abnormal metabolic and pathophysiological changes. Female breeders developed similar pathophysiological changes after four pregnancies, irrespective of their paired or communal breeding environment. Virgin rats were normal regardless of housing conditions. Our findings suggest that repeated breeding in male and female rats causes resetting of the hypothalamic-pituitary-adrenal-gonadal axis. This may lead to disturbed hormonal and metabolic changes which culminate with the development of accelerated cardiovascular degenerative changes.


The role of canine superficial ventricular muscle fibers in endocardial impulse distribution. Thin sections of canine right and left ventricular endocardium and myocardium were studied in a tissue bath to compare conduction properties of intraventricular specialized conducting tissue [Purkinje fibers (PF)], the superficial layers of subendocardial ventricular muscle (SVM), and the deeper ventricular muscle (DVM) below this level. The study was carried out because of observations that some areas of the endocardium, which are devoid of either specialized conducting tissue or of PF-VM junctions between specialized conducting tissue and ventricular muscle, conduct relatively rapidly, favoring specific orientations of propagation. Preparations containing PF, SVM, and DVM were studied electrophysiologically and histologically. A technique of stripping limited areas of endocardium was used to expose DVM in order to determine its intrinsic calculated conduction velocity. In 12 preparations, the average calculated conduction velocity in PF was 1.62 m/sec, and the average in DVM was 0.26 m/sec. The SVM conduction velocity was intermediate between the two, averaging 0.98 m/sec when propagation was parallel to SVM fiber orientation. Conduction velocity transverse to SVM fiber orientation was not significantly different from DVM conduction velocity. Histologically, the most superficial layers of VM were oriented uniformly in the direction of rapid subendocardial conduction, in contrast to DVM fibers in which orientation varied. It is concluded that the geometric arrangement of SVM fibers may provide a means for rapid subendocardial conduction and impulse distribution at a conduction velocity intermediate between PF and DVM in areas devoid of specialized conducting tissue.


Regional distribution of blood flow during arterial hypertension produced by lesions of the nucleus tractus solitarii in rats. Changes in the fractional distribution of cardiac output (FF), organ blood flow, and regional vascular resistance were measured by the isotope dilution technique of Sapirstein using 86Rb as indicator in unanesthetized rats during acute arterial hypertension produced by bilateral lesions of the nucleus tractus solitarii (NTS). After NTS lesions, the FF was significantly reduced in skin, muscle, and colon, increased in ventricular myocardium, spleen, and adrenal glands, and was unchanged elsewhere. Because of a marked reduction in cardiac output (CO) during hypertension, the absolute organ blood flow (FF X CO) was reduced in lesioned rats to 30-40% of control in skin, muscle, and colon and between 60% and 75% of control in most of the remainder of the gastrointestinal tract and renal cortex; it was unchanged in myocardium and endocrine glands. Resistance was substantially increased (4- to 6-fold) in skin, muscle and colon but was only moderately increased (1.5- to 2.5-fold) in the remaining organs. The results indicate that, while NTS lesions will increase resistance in most vascular beds, the response is unequally distributed, influencing skin, muscle, and colon disproportionately to other tissues. Because of an interaction between a reduction in CO and little autoregulation, blood flow is reduced primarily in skin, muscle, and colon. The pattern of redistribution of CO was consistent with the interpretation that NTS hypertension results from interrupting baroreceptor reflexes centrally. The pattern of redistribution of blood flow in rats with NTS lesions differs from that produced by deoxycorticosterone acetate-salt and renal ischemia.


A trial of ICI 74,917 in seasonal allergic rhinitis. Twenty-four patients with seasonal allergic rhinitis were treated in a randomized double-blind trial of 6 weeks' duration, using either ICI 74,917 (0.5 mg in each nostril four times a day) or an identical placebo aerosol. Four patients receiving placebo were withdrawn from the trial because of the deteriorationin their rhinitis. The twelve patients using ICI 74,917 were protected as assessed by a combined total symptom score, which was statistically significant (P less than 0.05), from weeks 1 to 4 when the pollen counts were highest.


A study of serum antibodies to isolated milk proteins and ovalbumin in infants and children. Serum antibodies to cow milk proteins and ovalbumin were measured quantitatively. Food hypersensitivity of the immediate type was determined to be present or absent by skin tests and double-blind food challenges. Elevated levels of antibodies to milk proteins in sera characteristic of infants fed cow milk were found to decline with age, so that sera from children who were 6 to 15 years of age (inclusive), not hypersensitive to food, had significantly lower levels than the infants. In contrast, sera from age-matched children, who were shown to have hypersensitivity to some food, were found to have levels of antibodies to milk proteins as elevated as in infancy. Hypersensitivity was not necessarily to milk but often to some other food. This persistence of greater antibody production to milk throughout childhood in those hypersensitive to some food indicates a fundamental difference from those without hypersensitivity to food, either in permeability, in immunological reactivity of the gut or in development of immunological unresponsiveness. Implications for pathogenesis of clinical disorders are discussed.


A simple method for the measurement of low levels of serum IgE. A simple method for the estimation of low serum IgE levels is presented. Radioimmunoassay often gives falsely high values because of the presence of non-specific factors in the serum when an excessive amount of serum is used in order to measure low serum IgE levels. Therefore the serum sample has to be diluted to minimize the influence of the non-specific factors and the lowest IgE level measurable in the serum is about 5 u/ml. In this study, pretreatment of the serum with cellulose powder was found to remove the non-specific factors that yield falsely high values. These factors were adsorbed almost completely onto cellulose powder when the serum IgE level was less than 100 u/ml. Thus, low serum IgE levels could be determined by radioimmunoassay using a sufficient amount of serum pretreated by cellulose powder. The lowest serum IgE level measurable by this method was as low as 0.5 u/ml and the low IgE level in neonatal serum could be determined with sufficient accuracy. By this method the geometric level of IgE in cord serum was 2.45 u/ml with a +/- 2 standard deviations range of 1.04 to 5.80 u/ml.


Evaluation of homogeneous enzyme immunoassay of serum thyroxine with use of bichromatic analysis. We evaluated the homogeneous enzyme immunoassay ("EMIT") of serum thyroxine with use of an ABA- 100 Bichromatic Analyzer, modified with an auxiliary dispenser syringe and a dual probe assembly. The modification allows two reagents to be dispensed at one time. The sera must be pre-treated with 0.1 mol/liter NaOH. The procedure thereafter is entirely automated. Twenty-four patients' samples may be analyzed in 15 min of instrument time. Precision and accuracy are excellent, and results correlate well with those by radioimmunoassay.


Changes in serum and urine lysozyme activity after kidney transplantation: influence of graft function and therapy with azathioprine. We investigated changes in lysozyme activity in serum and urine of kidney-transplant patients, and found that the production and catabolism of lysozyme in such patients differs markedly from that in normal subjects. Resumption of graft function decreases the high serum lysozyme activity by increasing the rate of catabolism in the transplant; at the same time, however, the production is inhibited by therapy with azathioprine. Changes in serum lysozyme activity correlate well with leukocyte count; thus its determination might be useful in monitoring immunosuppression. The urinary excretion of the enzyme, although not specific to rejection, is a good index of the degree of tubular damage.


Homogeneous enzyme immunoassay for cannabinoids in urine. We describe a homogeneous enzyme immunoassay for measurement of cannabinoid metabolites, as well as delta9-tetrahydrocannabinol (I) in urine. Malate dehydrogenase from pig heart mitochondria was labeled with a derivative of I. The compound used to calibrate the assay was the I metabolite, 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid (II). With 15 microgram of II per liter of urine as the cutoff concentration, the assay can detect 25 microgram of II per liter with greater than 95% confidence. A positive response was obtained for urine specimens assayed within 30 min after exposure to cannabinoids. However, the persistence of metabolites of I in urine indicates that assay of this fluid is useful as an indicator of cannabinoid use but not as an indicator of intoxication.


A rapid and sensitive gas-liquid chromatographic procedure for the micro determination of sodium valproate (sodium di-N-propylacetate) in plasma or serum. A gas-liquid chromatographic procedure for the micro determination of sodium valproate is described. Valproic acid is extracted from acidified plasma or serum into n-heptane containing an internal standard (octanoic acid) and after phase separation an aliquot is analysed by gas-liquid chromatography. The procedure is rapid, reliable, sensitive and specific. It requires 25--50 microliter sample for a single estimation, has a detection threshold of less than 10 mumol/l, and is suitable for routine clinical use.


Serum zinc levels and urinary zinc excretion in patients with renal transplants. An investigation into serum and plasma zinc levels and the urinary excretion of zinc in renal allotransplant patients was undertaken. A remarkable number (53%) of low serum and plasma levels of zinc was obtained as well as a distinct increase (37%) in urinary excretion in the 33 patients investigated. Because of the general tendency in these cases to depletion of body stores of zinc, oral supplementation may prove of value.


Study of the effects of bromocriptine on sexual impotence. Serum testosterone, prolactin, LH, FSH and plasma 17 beta-oestradiol levels were determined in forty-seven male patients with sexual impotence. Low testosterone values and slightly elevated prolactin levels were observed in 19% and in 17% of cases, respectively. Since sexual function in the male seems to be controlled by both dopaminergic stimulatory and serotoninergic inhibitory mechanisms, bromocriptine was given orally to seventeen patients. As the preliminary results appeared encouraging, a double blind study was undertaken in the other thirty patients. No appreciable difference in hormonal pattern was noticed between bromocriptine and placebo treatment. As far as sexual function was concerned, good results were obtained in 52% of cases treated with bromocriptine and in 44% of patients given placebo.


Positive feedback of oestrogen on LH secretion in women in neuroleptic drugs. The effect of a-adrenergic and dopaminergic blockade on the ability of pituitary-hypothalamic axis to produce an LH surge in response to oestrogen, was studied in eleven women treated with the neuroleptic agents haloperidol, fluphenazine or chlorpromazine. Seven women on no medication were controls. 6.6 mg of oestradiol benzoate (OB) was injected i.m. after control serum was obtained and daily serum samples were obtained for four additional days. Sera were assayed for total oestrogens, LH, FSH and prolactin. Six of seven women in the control group and eight of eleven women treated with neuroleptic agents had LH surges to OB challenge. The mean prolactin of the treated women was 33.1 +/- 3.6(SEM) ng/ml and the mean prolactin of the controls was 14.9 +/- 1.5(SEM) ng/ml, P less than 0.005). Elevated levels of circulating prolactin did not prevent an LH surge to OB challenge. These results indicate that even in subjects with catecholamine blockade, an LH surge can be induced. This contrasts to results obtained in rats and suggests that the control mechanism of LH surge production differs in rodents and humans.


Catecholamine release from the adrenal medulla. Chromaffin cells in the adrenal medulla are specialized for the synthesis, storage, and secretion of catecholamines. These cells are innervated by preganglionic sympathetic neurons in the splanchnic nerves, and, because of their unique blood supply, are exposed to unusually high concentrations of glucocorticoids in the venous drainage from the adrenal cortex. Splanchnic nerve stimulation appears to be the most important determinant of adrenomedullary function. Chromaffin cells synthesize catecholamines from tyrosine. Splanchnic nerve stimulation leads to an increase in the activity of several of the catecholamine biosynthetic enzymes, and to an increase in the rate of catecholamine biosynthesis. Glucocorticoids cause the induction of the enzyme noradrenaline N-methyltransferase, and so are particularly important for the synthesis of epinephrine. Catecholamines are stored, together with ATP, Ca2+, and protein, in secretory vesicles known as chromaffin granules. Splanchnic nerve stimulation is the physiological stimulus for catecholamine secretion. Stimulation of the splanchnic nerves results in the release of ACh from nerve endings in the adrenal medulla. ACh causes an increase in the permeability of the chromaffin cells to Ca2+, and thereby leads to the entry of Ca2+ into the cells. Ca2+ then causes the secretion of catecholamines and of other chromaffin granule constituents from  the chromaffin cells by exocytosis. The biochemical mechanisms of exocytosis, and the mechanism by which Ca2+ stimulates this process, are still unknown.


Circulating T-cell numbers and their mitogenic potential in leprosy--correlation with mycobacterial load. The effect of treatment and mycobacterial load on circulating T-cell numbers and their functional ability was investigated in forty-one patients with leprosy. Both early binding T cells and their responses to phytohaemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM) were profoundly and uniformly depressed in untreated, and partially treated, bacilleferous lepromatous leprosy (LL) patients as compared with normal subjects and tuberculoid patients. On elimination of mycobacteria, subsequent to chemotherapy, LL patients regain normality in T-cell numbers and their functions. On the other hand, the specific response of lymphocytes to M. leprae did not alter with decrease in mycobacterial load. It appears that the decrease in T-cell numbers and the deficit in their mitogenic potential is a secondary consequence of disease and is related to the antigenic load in patients with lepromatous leprosy.


The detection of membrane and cytoplasmic immunoglobulins in human leucocytes by immunoperoxidase staining. A sensitive immunoperoxidase technique for the detection of immunoglobulin (the peroxidase--anti-peroxidase or PAP procedure) has been applied to fixed smears of normal human white cells. IgM was detected in approximately 5% of lymphocytes from normal donors. Most positive cells showed a characteristic 'hairy' peripheral staining pattern; a similar morphological appearance was seen in samples stained for IgD. The membrane (rather than cytoplasmic) localization of this IgM was inferred from the redistribution of staining induced by preliminary incubation of cell suspensions with anti-mu antisera before smearing and staining. B cell-depleted and B cell-enriched suspensions showed, respectively, reduced and increased percentages of IgM-positive cells. IgG was detectable in approximately 25% of normal lymphoid cells. In contrast to the IgM and IgD reaction patterns, these cells commonly showed a discontinuous distribution of reactivity, often localized to the cell uropod or to small cytoplasmic vesicles. However, when cells were prepared at 0 degree C, staining tended to be diffuse. These findings suggested that the PAP procedure was detecting Fc receptor-bearing lymphoid cells which had bound serum IgG. IgG was also demonstrated in normal polymorphs and monocytes. The specificity of this reaction was confirmed by the use of immunoabsorbant-purified antibodies. The possible practical advantages of this immunoperoxidase procedure for the detection of leucocyte immunoglobulin are considered, and the relevance of the demonstration of IgG in non-lymphoid cells to recent reports of this immunoglobulin in Hodgkin's disease and malignant 'reticulum' cells is briefly discussed.


Immunofluorescence studies for immunoglobulins and complement C3 in synovial joint membranes in psoriatic arthritis. Synovial tissues from fifteen patients with psoriatic arthritis were investigated with direct immunofluorescence staining for immunoglobulins (IgG, IgA, and IgM) and from eigth patients for complement component C3. As control groups, there were synovial tissues from seven patients with seropositive rheumatoid arthritis and five patients with meniscal tears. In psoriatic arthritis, immunoglobulins were found in plasma cells in 93% of the cases, always with the presence of IgG (93%) but also with IgA (47%) and IgM (7%). C3 could not be demonstrated. In seropositive rheumatoid arthritis IgG was demonstrated in all patients (100%), often together with IgA (43%) and IgM (57%). C3 was found in all of these patients. In patients with meniscal tears neither immunoglobulins nor C3 could be found. The present findings indicate immunological activity in synovial membranes in psoriatic arthritis. The low amount of IgM and the lack of C3 suggest a difference compared to seropositive rheumatoid arthritis.


In vitro cell response of Treponema pallidum-infected rabbits. I. Lymphocyte transformation. Lymphocyte response to phytohaemagglutinin (PHA), Treponema pallidum, cultivable Reiter strain and cardiolipin reagent were examined using peripheral lymphocytes of T. pallidum-infected rabbits and control animals injected intratesticularly with saline extract of normal rabbit testes mixed with T. pallidum-free saline extract of infected testes. The response to PHA was significantly suppressed only in the infected animals, reaching the lowest level at 30 days after infection. A substantial increase of lymphocyte stimulation in the presence of cardiolipin reagent at 20 days after infection correlated well with the appearance of the maximum titre of Wassermann antibodies. Significant (P less than 0.02) increase in spontaneous response of cells incubated for 5 days made the response to T. pallidum and the Reiter strain difficult to interpret. An explanation for the spontaneous response is offered.


Hypoglycaemic syndromes and antibodies to pancreatic islet cells. The sera of ten patients with unexplained hypoglycaemia were examined for antibodies to pancreatic islets. Antibodies to pancreatic A, B and D cells (ICAb) were detected in one patient with an associated gastrointestinal tumour.


Glomerulonephritis due to transplant rejection affecting a patient's own kidneys. The kidneys of a patient were removed 3 months after a cadaveric allograft; histological examination revealed tuberculosis and malignant nephroangiosclerosis. When examined one year later, the graft exhibited lesions of glomerulonephritis and of vascular rejection. Immunoglobulin deposits were shown by immunofluorescence in the patient's own kidneys as well as in the graft; they were isolated by acid elution and in both cases had lymphocytotoxic activity in vitro. It is suggested that the target organ released histocompatibility antigen with formation of antigen antibody complexes which deposited in the patient's own kidneys.


Acute renal failure as a complication of hypertonic saline abortion in a kidney allograft recipient. Acute renal failure following abortion by intra-amniotic hypertonic saline administration has been described only occasionally. This report concerns a patient with end-stage renal failure who was successfully treated with a kidney allograft and developed reversible acute renal failure following the termination of her pregnancy by intra-amniotic infusion of saline. We suspect that the combination of hemoglobinuria and low grade intravascular coagulation might have been a contributing factor in the development of renal insufficiency in this patient. To our knowledge this modality of abortion has not been used previously in pregnant women with transplanted kidneys. From our experience with one patient it seems wise to express a word of caution on the use of this technique in patients with a functioning kidney allograft.


The carriers of native vitamin B12 in normal human serum. 1. After fractionation of the vitamin B12-binding proteins of ten normal sera the components containing transcobalamin II and R-type binders of vitamin B12 respectively were studied for endogenous vitamin B12 content by two distinct systems of vitamin B12 assay. 2. The measurements of total serum vitamin B12 by either bioassay with Euglena gracilis or a radioisotope dilution assay agreed closely. 3. The native vitamin B12 carried by transcobalamin II was higher as measured by bioassay than by isotope dilution assay. 4. The presence of the transcobalamin II fraction of human serum altered the key reaction between the binding reagent of the isotope dilution assay, so that this assay failed to measure vitamin B12 quantitatively. 5. Probably, the mean fraction of plasma vitamin B12 carried by transcobalamin II is in the range 20--30%.


Oral absorption of pivampicillin and ampicillin in young children: cross-over study using equimolar doses of a suspension. In a cross-over study, the oral absorption following equimolar doses of ampicillin corresponding to 16.7 mg/kg administrered as pivampicillin ('Pondocillin') suspension and ampicillin ('Doktacillin') suspension respectively, was investigated in 11 children aged from 8 months to 4.5 years. Pivampicillin produced a mean peak serum concentration of 10.7 microgram ampicillin/ml compared with 5.8 microgram/ml after ampicillin. The mean area under the serum concentration-time curves was 29.2 microgram/ml-h after pivampicillin and 16.7 microgram/ml-h after ampicillin, reflecting the superior bioavailability of the ester form. In 4 of the children aged between 8 and 12 months, the serum levels after pivampicillin were consistenly lower than those recorded in the remaining 7 children beyound the age of 1 year. The mechanism and therapeutic implications of this finding require further study.


The nutritional and physiological impact of cereal products in human nutrition. There is a need for reeducation of the population, especially in developed countries, as to the value of cereals in the diet. Cereals provide calories and important nutrients to the diet. Refined cereal products and unrefined cereals have certain advantages and disadvantages. With refinement, some nutrients and fiber are removed, but the body is better able to make use of certain nutrients. Essential nutrients are being replaced through fortification to compensate for losses in processing. The high fiber content of unrefined cereal products is believed to aid in the prevention of certain diseases. Special dietary bakery products have been introduced for the treatment of conditions generally exacerbated by standard food items. The increased consumption of cereal products appears warranted as a means of decreasing the saturated fat and cholesterol consumption. Cereals and cereal products have been mentioned in connection with allergies, celiac disease, schizophrenia, obesity, dental caries, cancer, atherosclerosis, goiter, and diverticulosis. This review discusses the possible role of cereals in the prevention or cause of these health problems.


Local treatment of psoriasis with desoxymethasone and betamethasone 17,21-dipropionate: a double-blind comparison. A double-blind comparison of the effectiveness of 0.25% desoxymethasone and 0.05% betamethasone 17,21-dipropionate creams was carried out in 40 patients with symmetrical, chronic psoriatic lesions. The lesions were pre-treated for 1 week with an inactive cream base and then the trial preparations were applied, without occlusion, to one or other side at random twice daily for 21 days. Overall response to treatment and the effect of the two topical steroids on scaling, induration, erythema, and pruritus were assessed at the start of and 4, 7, 14, and 21 days after the start of active treatment. The results indicated a better but not statistically significant response to desoxy-methasone. By the end of the trial period, the desoxymethasone-treated side was better in 22.5% of cases compared with 10% of cases in the betamethasone dipropionate-treated side. No side-effects of treatment were observed.


Controlled telethermographic study of the peripheral vasoactivity of oral piribedil. A single-blind crossover study was carried out in 10 subjects with a healthy arterial system to compare the effects of oral piribedil and placebo on the peripheral circulation. Using a telethermographic technique, skin temperature variations from baseline values were measured, at 15 minute intervals over a 2-hour period, after treatment with 3 x 20 mg piribedil tablets, after 1 x 50 mg piribedil in a sustained-release tablet formulation, and after placebo. In contrast to placebo, a vaso-active effect was observed afte piribedil administration on all but 3 occasions. Peak temperature changes appeared later after the sustained-release tablet. Side-effects of treatment were minimal and no significant changes were recorded in blood pressure.


A double-blind trial of feprazone in osteoarthritis of the hip. A double-blind crossover study was carried out in 21 patients with osteoarthrosis of the hip to compare the efficacy and tolerance of feprazone (600 mg/day) and ibuprofen (1200 mg/day), each drug being given for 4 weeks. No statistically significant changes were noted in any of the objective parameters measured, but patients' subjective assessments of pain showed a significant improvement in pain levels (p less than or equal to 0.05, day and night) after feprazone. One patient, who had reported a rash at initial assessment, was withdrawn at the end of the first treatment period (on feprazone) when he developed a severe rash, 1 patient was withdrawn because of exacerbation of symptoms (on ibuprofen) and  a further patient was lost to follow-up because of intercurrent illness. Both drugs were well tolerated by the other patients and the few side-effects reported were minor in nature.


Treatment of fluid retention in cirrhosis: a comparison of bumetanide and frusemide. Bumetanide and frusemide were compared in a crossover study of 10 patients with cirrhosis and fluid overload. Patients received each drug for 3 months. Doses of bumetide varied from 1 mg on alternate days to 3 mg daily (mean 1.3 mg/day), and for frusemide from 40 mg on alternate days to 160 mg daily (mean 72 mg/day). Both drugs proved effective in controlling ascites and oedema, 9 out of the 10 patients showing a satisfactory response. Minor side-effects, hypokalaemia and hyperuricaemia were common with both agents, and hypomagnesaemia and metabolic alkalosis developed in some patients.


Synergism of polyene antibiotics with 5-fluorocytosine. The synergism of 5-FC with 4 polyene antibiotics, amphotericin B (amph. B), candicidin, trichomycin and nystatin were investigated in 3 in vitro models measuring the fungistatic and fungicidal activity as well as the development of resistant mutants. In these 3 models candicidin and trichomycin exerted a higher synergistic effect than amph. B or nystatin. In vivo (systemic treatment of septicemic candidiasis of the mouse) the combination of 5-FC with amph. B was, however, the most effective. Only with this combination a complete cure (culturally negative) was observed. For topical treatment of Candida vaginitis in rats, the combination of 5-FC with candicidin proved the most active. As far as the biochemical basis of synergism is concerned in Candida albicans in the presence of polyene antibiotics, the incorporation of fluorinated pyrimidines was increased and the reduction of uptake of histidine by 5-FC alone, significantly enhanced. These effects could not be observed in Cryptococcus neoformans. The release of amino acids, phosphate and potassium caused by polyenes was significantly more pronounced in cells pretreated with 5-FC. Thus, the interaction between 5-FC and the polyenes may, in fact be, mutual.


Binding of chloramphenicol and its acetylated derivatives to Escherichia coli ribosomal subunits. Chloramphenicol-acetylated derivatives (1-monoacetoxy-; 3-monoacetoxy-; 1,3-diacetoxy-chloramphenicol), produced by a wild type Escherichia coli strain carrying an R factor, have been extracted and purified. None of the acetylated derivatives has been found to bind to E. coli ribosomal subunits.


Bladder instillations with trimethoprim-sulfamethoxazole in the treatment of urinary infection. 205 paraplegic inpatients with urinary infections were treated with trimethoprim-sulfamethoxazole (TMP-SMZ) bladder instillations (160 mg TMP 800 mg SMZ daily for 2 consecutive weeks). 89.3% out of 84 patients with recent vertebral trauma and no history of previous urinary infection (group A), and 68.6% out of 121 patients with paraplegia since more than 2 years, severe chronic infection and various pathological conditions of the urinary tract (group B) were found to be sterile at the end of the trial. The intravesical concentrations of 3.2 mg/ml TMP and 16 mg/ml SMZ which are reached immediately after instillation are 50--100 times superior to the minimal growth-inhibitory concentrations for most sensitive germs as determined with agar dilution tests. Several strains of Klebsiella and Proteus which had previously proved resistant to oral treatment were eliminated with intravesical TMP-SMZ. The incidence of reinfection, evaluated 1 month after interruption of treatment, was particularly high (62.7%) in patients with known involvement of the upper tract of intravesical obstruction with postmicturition residuals above 75 ml. On the other hand, 28% only among the patients with recent spinal injury and integrity of the upper urinary tract became reinfected after the same interval. The absence of local intolerance could be confirmed in all 21 cystoscopically examined patients between the 6th and 11th day of treatment. Hematuria was never encountered. Systemic side effects were noted in 7.8% Serum concentrations of both drug components are being investigated; results will be published elsewhere.


Group B streptococcal meningitis: delayed response to treatment. Four infants with group B streptococcal meningitis had a delayed response to antibiotic therapy. Resolution of cerebrospinal fluid infection and/or pleocytosis occurred only after prolonged and intensive antimicrobial management. In this respect, these infants were similar to infants with gram-negative enteric meningitis. It is suggested that infants with group B streptococcal meningitis be closely monitored to insure adequate response to therapy.


Relative efficacy of parenteral haloperidol and thiothixene for the emergency treatment of acutely excited and agitated patients. In this double-blind study, haloperidol (n = 15) and thiothixene (n = 15), administered parenterally in emergency rooms and outpatient facilities to 30 acutely excited, agitated psychotic patients in hourly doses of 4 mg. or 8 mg., as needed over a four-hour period (total dosage ranging from 4 to 32 mg.), achieved rapid tranquilization in 30 patients. Significant improvement was shown over a six-hour period on BPRS Total Score, the four factors--Thinking Disorder, Anergic state, Excitement and Disorientation, and Depression and also on hourly ratings of 17 symptoms of a Psychiatric Target Symptom Profile. No significant differences were found between the haloperidol-treated and thiothixene-treated groups. Few adverse reactions were noted, all of them mild, the most frequent being drowsiness in six patients.


[Renal transplantation in malignant nephrosclerosis (author's transl)]. Renal transplantation was performed 3 to 64 months after the onset of dialysis in 6 patients with histologically ascertained malignant nephrosclersosis and malignant hypertension, as well as terminal renal failure. Three patients were nephrectomized on both sides before and one after the transplantation. 24 to 105 months after the transplantation all patients were normotensive. Only one patient required low-dosage antihypertensive medication. There was evidence for marked improvement of cardiovascular complications, and no evidence for recurrence of malignant nephrosclerosis in the transplanted kidney.


Benzbromarone: a review of its pharmacological properties and therapeutic use in gout and hyperuricaemia. Benzbromarone1 is a benzofuran derivative which lowers serum urate and increases urinary urate excretion in normal, hyperuricaemic and gouty subjects. In open short- and long-term studies benzbromarone reduced serum uric acid levels by one-third to one-half and maintained its effectiveness for periods of up to 8 years. Single-dose experimental studies have shown benzbromarone to have a urate-lowering effect similar to that of a therapeutic dose of probenecid or sulphinpyrazone, but unlike these drugs benzbromarone can be administered in a once daily regimen. In 2 short-term comparative therapeutic trials in a small number of patients with hyperuricaemia, 80mg of micronised benzbromarone daily was at least as effective as 1000mg of probenecid or 300mg of allopurinol daily in lowering serum uric acid levels. Side-effects during benzbromarone administration are usually mild and primarily gastrointestinal in nature.


[Experimental control of the dye dilution method of measuring telediastolic ventricular volume by the steps in the curve of indicator "washing"]. After a brief discussion on the basis of the dilution method for determination of telediastolic volume (TDV) of the ventricles, the authors described the results from their own experimental studies on 8 nonbred dogs, on whom 186 curves of indicator "washing" (of the dye "Cardiogreen" were recorded, presented on 2 tables. The results showed that the measurements should be made by means of the later steps of the curves. TDV and accordingly the relative stroke volume of both ventricles differed slightly. The smaller values, according to the authors, of TDV of the left ventricle in comparison with the right ventricle revealed that the size of the relative stroke voluem depended both on the mechanism of Starling and on the contractility of the ventricles.


Characterization of the uptake of the methylxanthines theophylline and caffeine in isolated pancreatic islets and their effect on D-glucose transport. The uptake of theophylline and caffeine was determined in isolated pancreatic islets employing a dual isotope procedure with sucrose as an extracellular marker. Islets rapidly accumulated caffeine and theophylline with apparent dissociation constants of approximately 23 and 6 mM, respectively. Theophylline inhibited the uptake of caffeine and caused displacement of caffein from islets. These results indicated a competition by theophylline and caffeine for a common site (binding and/or transport carrier). In addition, theophylline and caffeine inhibited D-glucose transport in a dose-dependent manner and within the limits of the experimental system, this inhibition appeared to be non-competitive. (Bu)2cAMP under similar experimental conditions exerted no effect on D-glucose transport. These results present evidence for a rapid uptake of theophylline and caffeine in pancreatic islets, which is compatible with their immediate cellular effects. In addition, these results demonstrate a direct effect by theophylline and caffeine on D-glucose transport which appears independent of their ability to alter intracellular cAMP levels.


Circadian periodicities of serum androgens, progesterone, gonadotropins and luteinizing hormone-releasing hormone in male rats: the effects of hypothalamic deafferentation, castration and adrenalectomy. Serum testosterone (T), dihydrotestosterone (DHT), progesterone (P), LH, FSH and luteinizing hormone-releasing hormone (LHRH) levels and the LHRH content in the medial basal hypothalamus (MBH) and preoptic area (POA) were estimated during a 24-h period. Circadian rhythms, temporally unrelated to each other, were evident in serum FSH and LHRH and the MBH LHRH content; serum LH fluctuated randomly. In addition, serum T and DHT demonstrated a parallel circadian pattern while serum T and P appeared inversely related. Serum P rhythm persisted in long-term castrate males. Without adversely affecting serum LH, anterior hypothalamic deafferentation abolished both serum T and P rhythms, whereas adrenalectomy obliterated only the serum T periodicity. These studies show that the POA-MBH complex and the adrenals play important roles in the circadian regulation of testicular secretions.


Sensitized effect of prestimulation on insulin release from the perifused rat islets with a slow-rise glucose stimulation. The sensitized effect of prestimulation with 16.7 mM glucose on insulin release with a slow-rise glucose stimulation from the perifused rat islets of Langerhans was studied, together with the kinetic analysis of insulin release, and the interrelationship between the prestimulation time and the maximal rate of insulin release. All dose-response relationships which were derived from the dynamics of insulin release from islets prestimulated over various time periods within 60 min, showed sigmoidal profiles. Kinetic analyses were performed with Lineweaver-Burk's and Hill's equations. The 30-min prestimulation significantly reduced Hill's constant (n) from 6.2 +/- 0.7 of the control to 3.7 +/- 0.6 (p less than 0.05) and also enhanced the logarithmic equilibrium constant (log K) from -5.4 +/- 0.6 mM-n to -3.7 +/- 0.6 mM-n (p less than 0.05). However, the Km value was almost the same as that of the control (7.3 +/- 0.5 mM). On the other hand, the 60-min prestimulation remarkably diminished the Km value and the maximal rate of insulin release to 5.3 +/- 0.4 mM (p less than 0.05) and 0.6 +/- 0.08 muU/ml/islet/min (p less than 0.005), respectively. The maximal rate of insulin release linearly increased in proportion to the prestimulation time within 30 min. In conclusion, these results suggested that there would be some regularity depending on the prestimulation time in the process of transmission of the insulin-releasing signal in the pancreatic B cell and the accumulation of insulin into the provisional pool such as the labile insulin.


Direct projection from the medial preoptic area to the median eminence of the cat. The projection from the medial preoptic area to the median eminence of the cat was clarified by electron microscopy. After placing the electrolytic lesion in the preoptic area several kinds of degenerating neuronal processes and terminals were observed in the external layer of the median eminence. The one was dark shrunk terminals containing dense cored vesicles, the other was the dark ones containing myeline figure-like structure. The relationship between catecholamine-containing nerve endings and RH/IH-containing endings in the external layer of the median eminence was discussed.


Correlation of hypothalamic LHRH, pituitary LH and plasma LH in developing rats. Hypothalamic LHRH, pituitary LH and plasma LH levels were measured in rats of both sexes from day 5-60 after birth. The content of hypothalamic LHRH was very high in one-week-old male and female rats. It declined gradually till day 17 in the female rat and sharply on day 10 in the male rat. Subsequently the content of hypothalamic LHRH increased and showed peak values on day 25 in the female rat and on day 45 in the male rat. It decreased markedly at respective times of puberty in both sexes (day 37 in the female rat and day 52-60 in the male rat). Results of the study suggest that maturation of hypothalamo-hypophyseal-axis proceeds in three distinct stages. Observations on days 17, 25 and 37 in the female rat and on days 5, 7, 10 and 22 in the male rat clearly show an inverse relationship between hypothalamic LHRH and plasma LH and a parallel relationship between pituitary and plasma LH. Marked decline in the content of hypothalamic LHRH at respective times of puberty in both sexes indicates that the release of threshold levels of LHRH from the hypothalamus may apparently be the event initiating the pubertal changes in rat.


The role of the pineal gland in the regulation of LH-release in rats with different types of the anovulatory syndrome. The effect of different doses of testosterone propionate was investigated in provoking the development of the constant estrous anovulatory (CEA) syndrome in the rat. A direct relationship was observed between the dose of neonatally administered androgen (NA) and the percentage occurrence of this syndrome. Pinealectomy and superior cervical sympathetic ganglionectomy elicited the development of marked thecal luteinization in the NA-CEA rat, but the formation of corpora lutea was limited after these operations. The efficacy of pinealectomy and ganglionectomy in provoking luteinization was inversely related to the dose of testosterone used for neonatal androgenization. The LH-RH sensitivity of the adenohypophysis to release LH was decreased in the NA animals, as well as in the light-induced CEA syndrome (LCE), whereas it was increased in those CEA rats in which this syndrome was provoked by frontal hypothalamic deafferentation (FHD). Pinealectomy and ganglionectomy were able to elicit ovulation and luteinization in the FHD animals, but were ineffective in the LCE and high dose NA rats. The results are discussed in relation to those reported by others, in an attempt to explain the multitude of dissociated effects.


Pattern of islet lysosomal enzyme activities and insulin secretory response. The pattern of lysosomal enzyme activities in isolated pancreatic islets was studied in 3 different strains of mice, NMRI, CBA, and C-57, and related to the in vivo insulin release following injection of the insulin scretagogues glucose and carbachol. It was observed that the relative specific activities among the islet enzymes studied did not show the same pattern in the different strains although beta-gluc-ronidase always displayed the lowest activity. Comparison between the strains revealed that acid phosphatase activity was of the same magnitude in all 3 strains. Islet activities of acid amyloglucosidase, beta-glucuronidase, and N-acetylglucosaminidase, however, were largest in NMRI, intermediate in CBA, and lowest in C-57. This activity pattern roughly correlated with the insulin secretory response to an intravenous injection of glucose, whereas insulin release induced by the cholinergic agonist carbachol was of similar magnitude in all strains.


[Lipopolysaccharides from thermosensitive mutants of Escherichia coli K12 (author's transl)]. Thermosensitive mutants of Escherichia coli K12 were grown at 30 degrees C and 40 degrees C. The serologic properties and the composition of their lipopolysaccharides were investigated. The inhibition of hemagglutination by the lipopolysaccharides of various mutant strains was tested against the anti-E. coli K12 CR34 system. An inhibition was observed with all the mutants but one, CR34 T83 which had no inhibitory effect. Chemical analysis of lipopolysaccharides and mass spectrometric analysis of their methylated derivatives indicated the presence of the same components in the various lipopolysaccharides: glucose, galactopyranose, galactofuranose, heptopyranose and heptofuranose. However the 2,3,4 tri-O-methyl glucose is missing in the lipopolysaccharide of the mutant T83. This result agrees with the absence of a substituent on the 6-position of the non-reducing core-terminal glucose. The lipopolysaccharide of the T83 mutant has the complete core-type of E. coli K12. The relations of mutations with modifications of the composition of inner and outer envelopes in various mutants are discussed.


Enzymes synthesizing and hydrolyzing murein in Escherichia coli. Topographical distribution over the cell envelope. Envelopes from regions of the cell which in vivo show very little, if any, murein synthesis were isolated using the minicell-producing strain P678-54. Envelopes from minicells, representing in fact cell ends, were able to synthesize murein and to carry out transpeptidation in vitro; also all four murein hydrolase activities tested, carboxypeptidase, endopeptidase, amidase and transglycosylase, were found to be present. The specific activities of the murein synthesizing and degrading enzymes in envelopes derived from cell poles and from actively growing cells were similar. The topological distribution of murein-synthesizing enzymes and of murein hydrolases over the cell envelope is discussed.


Reconstitution of model membranes from phospholipid and outer membrane proteins of Proteus mirabilis. Role of proteins in the formation of hydrophilic pores and protection of membranes against detergents. Outer membrane proteins extracted from isolated cell walls of Proteus mirabilis were able to combine the cell wall phospholipids in a model membrane system. The presence of outer membrane proteins in vesicular model membranes mediated the release of previously entrapped [14C]sucrose while [3H]inulin was retained. Incorporation of lipopolysaccharide from the same cell walls was not required for the formation of such selectively permeable membranes. Three major outer membrane proteins of apparent molecular weights 39000, 36000 and 17000 were isolated using acetic acid and sodium deoxycholate solution as solvents and avoiding the strongly denaturing sodium dodecyl sulfate. The isolated proteins were assayed for their ability to form hydrophilic pores in reconstituted membranes. The trypsin-sensitive 39000-Mr protein and the peptidoglycan-associated 36000-Mr protein were equally effective in this function whereas the 17000-Mr protein mediated little penetration of low molecular weight solute. The 39000-Mr and 36000-Mr proteins also protected reconstituted membrane vesicles from disruption by detergent while 17000-Mr protein was ineffective in this regard.


Physico-chemical properties of a DNA binding protein: Escherichia coli factor H1. H1 factor is a heat-stable protein found in large amounts in Escherichia coli. In vitro, this protein has been found to stimulate transcription of lambda templates by E. coli DNA-dependent RNA polymerase. The subunit molecular weight of this factor has been re-estimated and found to be 15500 +/- 1000 in the presence of 2-mercaptoethanol. Crosslinking experiments performed with dimethylsuberimidate in the presence or absence of the DNA template indicate the presence of multiples of the 15500-Mr subunit up to the tetramer, the dimeric species being predominant. One cysteinyl residue per 15000-Mr subunit is labeled by 4-chloro-7-nitrobenzofurazan. This residue is not labeled if the factor is exposed to oxidizing conditions. In this case, three lysyl residues are titrated. H1 factor behaves as a DNA-binding protein. We have detected binding to DNA by two independent methods: displacement of a fluorescently labeled factor by the native protein and retention of radioactive DNA on millipore filters in the presence of the factor. Under our experimental conditions (high ionic strength, absence of magnesium ions), the saturation function of lambda plac DNA as well as of wild-type lambda DNA has been found to be non-cooperative. Saturation is reached when 300 +/- 30 molecules of dimeric factor are bound per lambda molecule, the average dissociation constant of the complex being 10nM. The dissociation time of the H1.DNA complex is less than 5 s at 37 degrees C. The binding of this factor lowers the affinity of native DNA for ethidium bromide. In the presence of this intercalating dye, the solubility of the complex decreases drastically.


Design of a study to evaluate drug therapy of serious ventricular rhythm disturbances after an acute myocardial infarction. A study was designed to investigate whether long-term use of aprindine can prevent sudden death from primary ventricular fibrillation. Patients with a proven recent myocardial infarction and malignant ventricular arrhythmias occurring late after the acute episode were asked to participate in a 1-yr, double-blind, randomized, placebo-controlled trial to suppress the rhythm disturbances observed on an ambulatory electrocardiogram. Particular care was taken to monitor drug adherence. Arrhythmia detection by ambulatory electrocardiography was used to assess drug efficacy; side-effects establish the maximum tollerated dose for each individual patient. Aprindine was therefore used under optimal circumstances. An interactive computer system served as a data base and provided the investigators and the monitoring committee with all the information required for a proper evaluation of the progress of the study.


Electrophysiologic evaluation of intravenous verapamil in man. The acute electrophysiological effects of intravenous verapamil (0.15 mg/kg body weight) were studied in 21 subjects with estimated normal impulse formation and conduction. Significant effects were sinus cycle shortening, depression of intranodal conduction and prolongation of AV node refractory periods. Sinus node recovery time, sinoatrial conduction time, atrial refractory periods, infranodal conduction, His--Purkinje system, and bundle branch refractory periods were unchanged. The clinical implications of these properties are discussed.


Reproducibility of diagnostic results by a multivariate computer ECG analysis program (AVA 3.5). Reproducibility of the AVA program (version 3.5) was tested (1) by analyzing the same analog tracing 10 times in 150 patients, and (2) by studying the influence of 12 different sets of prior probabilities (PRP), using the digital data from a total of 2718 patients. The same QRS-T diagnosis plus or minus 8 percentage points was found in patients with high posterior probabilities. Variability for the P wave diagnosis was twice as high as for QRS-T results. Failure to select the appropriate set of PrP markedly influenced reproducibility. This was especially true for right ventricular hypertrophy and pulmonary emphysema. 'Mispriorizing' in a subgroup of 199 patients with well-documented myocardial infarction lowered the total number of correct ECG diagnoses on the average between 4.8 and 35%. These results are inherent to Bayes' theorem where PrP are used to achieve minimum assignment error in populations, but where Prp errors may result in wide classification discrepancies. Without errors in PrP reproducibility by the VA program was found to be very high.


Rabbits possess conventional T lymphocytes. Rabbit lymphocytes have been analyzed as to surface Ig markers in relation to the function of the cells. A battery of specific anti-Ig reagents as well as supposed B and T cell-specific mitogens were used, and DNA synthesis as well as high-rate Ig synthesis in vitro were recorded. Using cells from spleen lymph node or blood, surface Ig-negative lymphocytes expressed the expected behavior of T lymphocytes. No evidence was found of significant expression of allotypic markers on the surface of such nonactivated rabbit T lymphocytes. Lymphoid cells from bone marrow constituted an exception to the rule in the sense that they contained a high proportion of cells being surface Ig-negative at the time of anti-Ig column fractionation. They did, however, rapidly express surface Ig molecules as well as B cell markers as judged by B cell mitogenic stimulation shortly after in vitro explanation. In conclusion, we failed to find any constant region Ig markers on rabbit lymphocytes, which in every sense behave like conventional T lymphocytes.


The "patchy" immunodeficiency of CBA/N mice. The in vivo responses of CBA/N (which have an X-linked defect of B lymphocyte differentiation) and CBA/J normal mice to 2,4,6-trinitrophenylated (TNP) bacteriophage T4 and Diplococcus pneumoniae CS variant have been compared. Both antigens are thymus-independent (TI), and TNP-CS additionally contains the phosphorylcholine (PC) epitope, CBA/N and CBA/J mice give TNP plaque-forming cell responses similar in magnitude and avidity although only CBA/J give a PC response when challenged with TNP-CS. These results indicate that CBA/N mice have a "patchy" defect of antigen-induced humoral responses. This defect is manifested only in certain subpopulations of B cells and is not restricted to TI responses.


Transformation of arachidonic acid and prostaglandin endoperoxides by the guinea pig heart. Formation of RCS and prostacyclin. The metabolism of arachidonic acid (AA) was studied in perfused isolated hearts from guinea pigs. The coronary effluent was continuously bioassayed for prostaglandin-like substances (PLS) using the cascade technique of Vane. Injections of AA in doses between 1--50 microgram into the perfusion fluid prior to the heart produced vasodilatation of the coronary vascular bed followed by a contraction of the rat stomach strip (RSS), chick rectum (CR) and rat colon (RC) as well as relaxation of the bovine coronary artery (BCA). At the higher doses of AA there was also contraction of the rabbit aorta (RbA). The same pattern of effects on the bioassay tissues was seen when prostaglandin endoperoxide (PGH2) was perfused through the heart. The response of the bank of superfused tissues provided evidence for the formation of prostacyclin (PGX or PGI2), PGE2 and PGF2alpha. Chromatographic studies showed that 6-oxo-PGF1alpha together with other prostaglandins was present in the perfusate after acidification, which suggested that the bovine coronary relaxing substance consists mainly of PGI2. Moreover, the rabbit aorta contracting substance (RCS) released in the perfusate was due to prostaglandin endoperoxides and not to thromboxane (TXA2). The formation of PLS from AA was completely blocked after treatment of the heart with the cyclo-oxygenase inhibitors, indomethacin or meclofenamic acid. Pretreatment of the heart with 15-hydroperoxyarachidonic acid (15-HPAA), a selective inhibitor of prostacyclin synthetase, inhibited the effect of AA on the coronary vasculature and diverted the metabolic transformation of AA towards PGE2 and PGF2alpha.


The role of the corpus striatum in neuroleptic- and narcotic-induced catalepsy. Lesion experiments (in the rat) were designed to elucidate the function of the corpus striatum in neuroleptic- and narcotic-induced catalepsy, respectively. Bilateral lesions of the corpus striatum were observed to attenuate neuroleptic (CPZ)-induced catalepsy. However, analogous lesions of the corpus striatum potentiated narcotic (morphine)-induced catalepsy. These results suggests that (a) the corpus striatum may be a primary site of action of neuroleptic drugs (such as CPZ) in the production of catalepsy, and (b) narcotic (morphine)-induced catalepsy may not be exclusively mediated by the corpus striatum.


The statistical distribution of colony counts in the spleen colony assay. The statistical distribution of colony counts in the spleen colony assay was determined in C57BL/6 and C3H mice under varying conditions of the assay procedure. Assay mice were given syngeneic bone marrow cells from normal or irradiated donors. The mice were injected intravenously with marrow cells either immediately or 3 days after irradiation. Eight days after engraftment, spleen colonies were counted and the statistical distribution of colony counts was determined using a mean to variance ratio criterion. Analysis of the data indicated that the statistical distribution of colony counts was different in the two strains of mice. In addition, delaying time of injection of mice with bone marrow cells or injecting bone marrow cells form irradiated donors also changed the statistical distribution. The significance of this work lies in the fact that modification of the usual assay procedure can alter the statistical distribution of colony counts, thus changing the statistical accuracy of the assay. Modification in the assay procedure, such that an increase occurs in the variance of the distribution of colony counts, must be compensated for by increasing the number of spleens counted in order to maintain a constant level of statistical accuracy in experiments.


Adenohypophysial necrosis in rats following destruction of the pituitary stalk. A histologic, immunocytologic and fine structural study. Adenohypophyses of adult male rats have been investigated by light microscopy, immunocytology and electron microscopy 30 minutes, 1, 2, 4, 6 and 24 hours following electrolyte destruction of the pituitary stalk. Light microscopy revealed massive ischemic infarction of the adenohypophysis. Immunoreactive growth hormone, prolactin, TSH, FSH and LH were demonstrated up to 24 hours after surgery in necrotic adenohypophysial cells by the immunoperoxidase technique. Fine structural abnormalities were already noticeable 30 minutes following stalk lesion. The changes rapidly progressed and within 2 hours unmistakable signs of cellular necrosis became evident. Mitochondria, especially those of growth hormone cells, seemed to be affected at an early stage, whereas alterations in rough-surfaced endoplasmic reticulum membranes developed later. Nuclear changes, formation of cytoplasmic vacuoles and disruption of cell membranes were conspicuous findings. No evidence, indicating the participation of lysosomes in the development of cellular damage, was obtained. Secretory granules were prominent and well preserved even in cells which showed advanced necrosis. The failure of discharge and degradation of secretory granules in the necrotic cells suggest that factors accounting for their intracellular migration and extrusion are very sensitive to ischemia and are paralyzed at an early phase.


A mutant of the antibiotic resistance factor R124 with altered copy number. A mutation conferring increased antibiotic resistance on Salmonella typhimurium strain 11G carrying R124 was plasmid determined; strains harbouring the mutant plasmid contained more DNA as ccc plasmid than those harbouring R124. The increased copy number was manifested at all growth rates tested.


Regulation of ammonia uptake in aspergillus nidulans. The ammonia uptake in A. nidulans was found to be linear for about 20 min, and was proportional up to 1.5 mg/ml dry cell density. The transport of ammonia does not involve energy. Normal and biotin deficient A. nidulans showed an identical Km-values of 10.26 X 10(-5) M ammonia for uptake. The uptake of ammonium ion has been shown to be regulated by the intracellular concentration of ammonia.


The use of fluorescein isothiocyanate labelled lectins for immuno-histological demonstration of saccharides. III. Studies by use of Ricinus communis lectin and wheat germ agglutinin. Cryostat sections of heart, skeletal muscle, spleen, liver, cartilage, brain, nerve, tongue, esophagus and salivary gland from man, rabbit, rat and guinea pig were stained with fluorescein isothiocyanate labelled Ricinus communis lectin and wheat germ agglutinin. The stained sections were examined by fluorescence microscopy. The general pattern of fluorescence is very similar to that obtained with labelled Lens culinaris lectin and Concanavalin A. Particularly the lectins stain connective tissue structures of the organs. The staining intensity in these structures was weaker by wheat germ agglutinin as compared to the other lectins.


Transcription of bacterial DNA by isolated plant nuclei. Plant nuclei prepared from protoplasts can be used as a cell-free system for testing their template activity of procaryotic DNA for plant polymerases. We were able to demonstrate that plant polymerases of Petunia hybrida are capable of transcribing linear bacterial DNA, whereas supercoiled DNA could not be used as a template.


Anti-A agglutinin from Helix pomatia as a marker for L cells. By means of fluorescein-labeled anti-AHP agglutinin isolated from the albumen gland of the snail Helix pomatia L. L cells were detected in air dried and fixed monolayer preparations, smear preparations or cryostat sections of tumours induced in two different host systems. The tumours consisting of L cells could be clearly distinguished from the surrounding normal host tissues, and also single tumour cells lying separtely in the host tissued were identified in the same unequivocal manner. The cells were marked by characteristic granular fluorescence caused by specific agglutinin binding within the cell membrane. With these results the previously expressed thesis that anti-AHP  agglutinin might be used as a marker for L cells is confirmed for L cells grown in suitable host systems, too.


In vivo effects of Escherichia coli endotoxin on sulfobromophthalein clearance in the guinea-pig. In vivo studies indicated that the primary effects of E. coli endotoxin on hepatic clearance of sulfobromophthalein were at the excretory level. Newborns were more sensitive to the LPS than older animals.


The family interactional perspective: a study and examination of the work of Don D. Jackson. This article presents an overview of the primary contributions of the late Don D. Jackson. It analyzes and attempts to unify the central concepts of what the first referred to as "conjoint family therapy." Emphasis in upon the theoretical components leading to the development of a behaviorally oriented, nontransference, focused-treatment format, labeled by the author as "family interactional psychotherapy."


Structure-activity correlations for antibacterial 1,5-benzodiazepine derivatives on gram-positive cells. The antibacterial activity of a set of 1,5-benzodiazepine derivatives against Staph, aureus and Strep, pyogenes has been correlated with structural features of the compounds examined. It is shown by means of substituent constants and regression analysis that the lipophilic character of the molecule is the most important factor  in determining its activity. Electronic and steric effects appear to be quite important.


Mechanisms for the biomethylation of metals and metalloids. The case of methylmercury pollution has demonstrated the profound importance of understanding biologically mediated transformation reactions that yield organometallic compounds with a high potential for bioaccumulation and toxicity. Toxic elements that form organometallic compounds, especially the metal-alkyls (e.g., methylmercury), deserve special concern. Most metal-alkyls are poisonous to the central nervous systems of higher organisms, and these compounds do accumulate in cells. Metal-alkyls that are stable in water, and that have been reported to be synthesized biologically, can be formed from the following toxic elements: Hg, Sn, As, Se, Te, Pd, Au, Tl and Pb. In this report we present details of the mechanisms for biological methylation of certain metals and metalloids with special emphasis on those elements that are widely dispersed in the biosphere. In addition we present preliminary results on the use of flourescence quenching techniques to determine cellular diffusion rates and partition coefficients for methylmercuric chloride.


Effects of metals on heme biosynthesis and metabolism. Studying the mechanism of interaction of metals with enzymes of the heme biosynthetic pathway has revealed a great deal of understanding of the regulation of the pathway and a possible insight into the toxicology of certain metals. Both positive and negative effects of metals on heme biosynthesis and catabolism can be demonstrated. The understanding of these interactions has given us a clearer picture of certain of the descriptive events known to occur in toxic states induced by these metals. Unfortunately, the studies have also driven us to appreciate the even more complex nature of the biosynthetic and metabolic pathways for heme than was originally hypothesized. It can be predicted that the next 5 years should lead to many more revelations and potential treatment modalities for toxic states induced by metals or genetic diseases where heme biosynthesis does not occur normally.


Contributions of nutrition science to anthropological research. Certain investigations of nutrition scientists have demonstrated the effects of cultural practices on nutrient intake. Anthropologists, too, have studied diets and food-related behavior, although until recently their interests infrequently included nutritional status. In the last decade, nutrition scientists' research helped stimulate the creation of an interdisciplinary specialty, nutritional anthropology, that combines techniques from both nutritional and anthropological science in the study of food use in relation to health. Practitioners of this expanding field make useful contributions to research in human nutrition through study of social aspects of the diet of individuals and groups. Their assistance can be particularly helpful in studies of populations in developing countries and of ethnic minorities in this country. Certain unanswered nutritional questions, which might more readily be solved by the combined efforts of nutritionists and nutritional anthropologists than by either professional alone, are presented.


[Electron microscopical observations on the curative effect of ursodesoxycholic acid in alloxan-induced pancreatic islet cell injury (author's transl)]. The preventive effect of ursodesoxycholic acid on pancreatic injury by alloxan (alloxan diabetes) has been reported by Watari, et al. (1976). In the following experiment, to pursue the findings further, ursodesoxycholic acid was used curatively for alloxan diabetes. A first group of animals (5 mice) were injected with alloxan (4 mg) twice at the fifth and tenth day. The second group (5 mice) was injected with ursodesoxycholic acid (0.2 mg each) for 14 days during the experiment in addition to the same alloxan dosage/frequency as the first group. A third group of animals (5 mice) served as the control. The animals were sacrificed on the 15th day and the blood sugar levels were examined, using commercial test paper. The pancreatic tissues were fixed in a mixture of 2.5% glutaraldehyde and 2% osmid acid solution, which was adjusted at pH 7.4 with a veronal acetate buffer; the osmotic pressure was also regulated by adding sucrose of 0.045 g/ml. Following dehydration using a series of alcohol concentrations, the tissues were embedded in Epon 812. Thin sections were cut with a Porter-Blum MT-2B ultramicrotome, stained with both uranyl acetate and a lead mixture, and then observed by electron microscopy. The results were as follows: The pancreatic islet cells, especially of B-cells in the first group of animals injected with alloxan only, were seriously damaged and contained myelinated mitochondria. Golgi apparatus, and an increasing number of autophagic vacuoles. Some B-cells revealed hydropic degeneration. Some B-granules changed into vacuoles after diacrine secretion. Pancreatic A-cells were increased in number and showed no cell injuries. On the other hand, the pancreatic B-cells of mice treated with both alloxan and ursodesoxycholic acid maintained almost normal fine structures. In summary, ursodesoxycholic acid has a curative effect on alloxan-induced pancreatic B-cell injury.


[Detection of the antithyroglobulin antibody precipitated with polyethylene glycol (PEG) (author's transl)]. It is well known that the antithyroglobulin (anti-Tg) antibody plays an important role in the pathogenesis of autoimmune thyroiditis (chronic thyroiditis). The anti-Tg antibody is detected by double diffusion in agar gel, the fluorescent antibody technique and the tanned red cell haemagglutination test (TRC). The most widely used method is the TRC test, but it is negative in about 30 percent of the patients with chronic thyroiditis. In this paper, we have reported the detection of the anti-Tg antibody in serum using a modified Farr's method. In our method, PEG was used instead of ammonium sulfate to precipitate the immune complex formed in vitro between labelled Tg and the autoantibody. Percent 125I-Tg precipitated was 4.7 +/- 3.1 percent in normal controls; 20.4 +/- 11.4 percent in TRC negative sera were detected by this method. A good correlation was found between TRC titer and percent 125I-Tg precipitated by the PEG method in patients with chronic thyroiditis. By this method, the anti-Tg antibody was also detected in the sera of rabbits immunised with human Tg earlier than that detected by the TRC and double diffusion tests. The sensitivity and simplicity of this method provide a useful tool in detecting the anti-Tg antibody in clinical as well as in experimental work.


Localized chronic pemphigoid. Report of a case, review of the literature and discussion of the relationship with cicatricial pemphigoid. A case of a chronic remittant blistering eruption on the left temple in a 62-year-old woman is described. Only after repeated biopsies did direct immunofluorescence (DIF) studies reveal a linear staining of the epidermal basement membrane with anti-IgG, C1q, C4, C3 and C5. In addition, heavy deposits of fibrin were observed. The importance of repeated DIF examination in initially negative cases has been stressed. Localized chronic pemphigoid should be considered as a distinct subform of the pemphigoid syndrome.


A controlled trial of baclofen in children with cerebral palsy. A double-blind crossover trial against placebo was conducted to assess the effects of the GABA derivative, baclofen, on the disabilities due to muscle spasticity in twenty children suffering from cerebral palsy. Baclofen performed very significantly better than placebo in reducing spasticity and significantly better than placebo in allowing both active and passive limb movements to be carried out. Notable improvement was also seen in scissoring. Side-effects were minimal and responded promptly to dose reduction. The evaluation of drug effects on muscle spasticity and the pharmacodynamics of baclofen are discussed. Recommendations are made regarding dosage of baclogen in childhood.


The effect of fenoprofen calcium (Nalfon) on levels of rheumatoid factor in patients with rheumatoid arthritis. In 88.9% of a group of 18 patients with rheumatoid arthritis participating in an open-label study with fenoprofen calcium (Nalfon), there was observed a mean titre decrease in level of rheumatoid factor as compared to baseline levels. In follow-up, three months to one year later, a uniform titre increase above the mean level of the study period had occurred. In all cases the titre returned to baseline levels or higher.


Pain relief in the post-operative period: a comparative trial of morphine and a new analgesic buprenorphine. In a comparative trial, buprenorphine 0-3 mg or morphine 10 mg was administered intramuscularly to patients post-operatively. The new drug buprenorphine produced more pain relief than morphine and appeared to have a longer duration of action. The side-effects produced by the two drugs were similar, as were the effect on respiratory and cardiovascular measurements.


Double-blind placebo controlled oral analgesic comparison of butorphanol andpentazocine in patients with moderate to severe post-operative pain. A double-blind, randomized trial was conducted in 120 post-surgical patients to evaluate the oral analgesic activity of butorphanol tartrate (4 mg and 8 mg) and pentazocine HCl (50 mg) as compared to placebo. Both dose of butorphanol as well as pentazocine proved to be significantly (p less than 0-05) more effective than placebo. Butorphanol 4 mg and pentazocine 50 mg were never significantly different from each other, while butorphanol 8 mg was significantly beeter than both butorphanol 4 mg as well as pentazocine 50 mg in several instances, demonstrating a significant dose effect relationship for butorphanol. All of the active treatment provided maximum pain relief within 1 to 2 hours and were effective over 4 hours. In contrast to the other treatments, none of the 8 mg butorphanol patients required remedication during the 4-hour observation period. Generally, the incidence fo side-effects appeared low.


Gentamicin in the treatment of staphylococcal infections. The aminoglycoside antibiotic, gentamicin, was used to treat staphylococcal infections in eighty-six patients in an open multicentre trial. Most of the infections involved the skin and soft tissue and the lower respiratory tract. Staphylococcus aureus was the only organism isolated in seventy-four patients; mixed flora were found in twelve. Gentamicin was administered, intramuscularly or intravenously, for 7 to 12 days (mean, 10 days) in a mean dose of 3-27 mg/kg per day. Clinical and bacteriological assessment of results indicated a complete resolution of the infection in fifty-three patients (61-6%) and a marked, moderate, or slight improvement in an additional twenty-nine patients (33.7%). Thus, a total of eighty-two patients ((95.3%) showed cure or improvement while only four patients (4-6%) failed to do so. Staphylococci persisted in six patients. Superinfection also occurred in six patients, however, it was considered to be clinically significant in only four of them. Screening for eighth cranial nerve, renal, hepatic and haematological function, before, during, and after gentamicin treatment, revealed no adverse reactions in these patients.


A double-blind clinical study with benzydamine 3% cream on soft tissue injuries in an occupational health centre. A double-blind, controlled clinical study with benzydamine hydrochloride 3% cream was conducted in an Occupational Health Centre on fifty patients with soft tissue injuries (two mild, ten moderate and thirteen severe cases in the placebo group and one mild, thirteen moderate and twelve severe cases in the active group). The trial demonstrated the significant therapeutic superiority of 3% benzydamine cream over inactive placebo in all six qualitative parameters for traumatic inflammatory changes at the end of a six-day assessment and a marked improvement of symptoms event two days after the onset of therapy. In addition, tbe incidental use of ultrasound therapy together with the standard treatment revealed the value of the active drug as an effective couplant. Neither group suffered any side-effects. Although the size of population studied was small, it was apparent that active treatment with benzydamine was a distinct aid to healing of soft tissue injuries and this should be confirmed with further large-scale clinical trials.


The transient anti-hypertensive effect of phentolamine in patients receiving beta-blocker treatment. In seven hypertensives receiving beta-blocker drugs, and additional reduction in standing blood pressure occurred between 60 and 90 minutes after 40 mg phentolamine by mouth. The occurrence of the postural hypotensive effect was delayed in relation to the reported time of peak plasma concentration of unchanged phentolamine. Supine blood pressure and heart rate were unaffected. Phentolamine has no clinically useful anti-hypertensive effect in conjunction with beta-blockers in patients with essential hypertension.


A trial of Lotussin and linctus diphenhydramine in patients wth an irritant cough. Lotussin and linctus diphenhydramine were compared for efficacy and patient preference in fifty patients suffering from post-infective cough. The trial was a single-blind randomized crossover one which was subjected to sequential analysis based on patient preferences. Under the conditions of the trial there appeared to be a significant difference in favour of Lotussin (p less than 0-01).


A comparison of maprotiline (Ludiomil) and amitriptyline (1). The study described here was designed to see if it was possible to show an earlier onset of action for maprotiline (Ludiomil) compared with amitriptyline while at the same time demonstrating equal long-term effectiveness and certainly no greater side-effects. A between-patient double-blind trial with 40 patients supported these aims and was statistically significant (p less than 0.001) with regard to early onset of antidepressant effect.


A comparison of maprotiline (Ludiomil) and amitriptyline (2). Seventy-five patients were admitted to a double-blind multicentre trial of maprotiline (Ludiomil) and amitriptyline in the management of depression in general practice. Forty-seven patients, twenty-one on maprotiline and twenty-six on amitriptyline completed the study. Statistical analysis of admission data showed that the groups were not strictly comparable in that the maprotiline group contained many more males and had a greater average age. Nevertheless, although there was a suggestion that amitriptyline was faster-acting in the younger age group that received it, there were no statistically significant differences between the two treatment groups as regards onset of effect, clinical efficacy or tolerability.


A comparison of high and low dosage regimes of maprotiline (Ludiomil). A double-blind, randomized, multicentre trial is described which was designed to compare the clinical effect of 30 mg and 75 mg daily of maprotiline (Ludiomil) administered as either once or thrice daily therapy. Physicians' assessment of patients' progress was made following 7, 14 and 28 days of treatment. Patients also assessed themselves using a visual analogue scale at the same time intervals. Of the 231 patients admitted to the study, 40 dropped out for various reasons, leaving 191 patients completing. Of the 40 drop-outs, drug-induced side-effects were considered to be responsible in the case of 17 patients. No differences between the four treatment groups were demonstrated on the physicians' assessment; however, the patients' self-assessments using the 10 cm visual analogue scale indicated that the 25 mg three times daily regime appeared to be the most satisfactory, but not statistically significantly so, when compared with the 10 mg thrice daily and 75 mg nocte regimes. The 30 mg nocte dose proved to be distinctly inferior.


A comparative study of maprotiline (Ludiomil) and viloxazine in the management of depressed patients in general practice. A double-blind, comparative, multicentre study comparing the efficacy and tolerability of maprotiline (Ludiomil) and viloxazine as antidepressants in general practice usage is described. Progress was measured at weekly intervals over the 4-week trial duration using a modified Hamilton Rating Scale and patient self-assessment visual analogue scales. A total of 127 patients were randomized into two groups; 63 patients being allocated maprotiline and 64 patients being allocated viloxazine. Of the patients receiving maprotiline, 12 withdrew during the course of the study compared to 9 patients receiving viloxazine. Maprotiline appeared to be superior to viloxazine on the sleep and sadness scales and some evidence to show it to be better also on the tension scale. Whilst the trend favoured maprotiline, the two drugs did not appear to be significantly different on either the doctors' or the patients' assessments. There was little to choose between the drugs in terms of side-effects or ultimate patient tolerability and preference.


Subcellular localization of the alterations in phosphatidylinositol metabolism following glucose-induced insulin release from rat pancreatic islets. The subcellular localization of the incorporation of 2-(3H)-myoinositol into lipids has been studied in isolated pancreatic islets of the rat. The recovery of lipid-bound myoinositol increased with time in the nuclear, mitochondrial, microsomal, and secretory granule fractions. The utilization of a filtration technique for the more complete separation of mitochondrial and secretory granule elements permitted us to show that the recovery of lipid-bound 2-(3H)-myoinositol increased most rapidly in the secretory granule fraction. A 30-minute exposure of prelabeled islets to a stimulatory concentration of D-glucose (3.0 mg./ml.) resulted in a statistically significant decrease in the amount of lipid-bound 2-(3H)-myoinositol that was recovered from the secretory granule fraction (p less than 0.001). In contrast, exposure of islets to the elevated glucose concentration had no statistically significant effect on the recovery of lipid-bound radioactivity from other subcellular fractions. Since the majority of lipid-bound radioactivity associated with the secretory granule fraction could be recovered with the presumptive secretory granule membranes, these data suggest that the hydrolysis of phosphatidylinositol that accompanies glucose-induced insulin secretion from the rat pancreatic islet may be localized to the beta granule and, in particular, to its limiting membrane.


Morphometric quantitation of the pancreatic insulin-, glucagon-, and somatostatin-positive cell populations in normal and alloxan-diabetic rats. The pancreatic insulin-, glucagon-, and somatostatin-positive cell populations were quantitated in normal and alloxan-diabetic rats. The method of quantitation (linear scanning) allowed an estimation of absolute changes in these cell populations through 14 months of diabetes. The changes in cell masses were correlated with changes in plasma and pancreatic immunoreactive insulin and glucagon. A marked reduction in the insulin-positive beta cells was demonstrated within seven days after alloxan treatment. No significant change in the glucagon-positive alpha cell population was noted in the diabetic rats when compared with normoglycemic controls. A statistically significant increase in the pancreatic somatostatin-positive delta cell population was demonstrable only after 14 months of alloxan diabetes. The results would suggest that the hyperglucagonemia of insulin-deficient diabetes is not a consequence of an increased pancreatic alpha cell population. In addition, since the increase in the pancreatic delta cell mass was found only late in the course of alloxan diabetes in the rat, the increase in delta cells is probably not of significance in the pathophysiology of diabetes in this experimental model.


Inhibition of glucagon release by serotonin in mouse pancreatic islets. In this work we have investigated the effect of serotonin on glucagon release in mouse pancreatic islets isolated by the collagenase technique. Incubation of the islets with serotonin (4 X10(-3)mol/l) was associated with an inhibition of glucagon output both in the basal medium (3.3 mmol/l glucose) and in the presence of arginine (10 mmol/l). The inhibitory effect of serotonin on basal glucagon release was also apparent at concentrations of 2 X10(-3) mol/l, 10(-3)mol/l and 5 X 10(-4) mol/l. Addition of 5-hydroxytrypophan (4 X10(-3) mol/l) to the incubation medium was without effect on basal glucagon output while it significantly reduced arginine-induced glucagon release. In contrast, tryptophan (4 X10(-3) mol/l) provoked glucagon secretion. As inferred from our previous human studies, the present data indicate that serotonin is able to inhibit glucagon secretion. These findings provide further support for the participation of a serotoninergic mechanism in the control of A-cell function.


Ninhydrin inhibition of glucose-induced insulin release. Ninhydrin, a compound which shares chemical properties strikingly similar to alloxan was found to mimic basically the inhibitory effect of alloxan on glucose-induced insulin release. Exposure of pancreatic islets for five minutes to 85 mumol/l ninhydrin produced approximately ninety percent inhibition of subsequent glucose-induced insulin release without altering basal secretion. Both D-glucose and D-mannose provided substantial protection against the inhibitory effect of ninhydrin, and the alpha anomer of D-glucose was more effective than the beta anomer in preventing ninhydrin inhibition of insulin release. Evidence for a common site of inhibition by ninhydrin and alloxan in the insulin release process is discussed.


Prevalence of residual B-cell function in insulin-treated diabetics evaluated by the plasma C-etide response to intravenous glucagon. In 83 insulin-treated diabetics the influence of the duration of insulin treatment on the prevalence of residual insulin secretion was examined by determining the plasma C-peptide concentration before and after intravenous injection of 1 mg of glucagon. In 64 patients, plasma C-etide concentration was also determined before and after a standard meal. There was a good correlation between the C-peptide response to glucagon and to the meal (r = 0.67; p less than 0.0001) suggesting that the glucagon test will predict the B-cell response during everyday life. The predictive value of a positive glucagon test was 84% and of a negative test 100%. A preserved, but reduced, B-cell function was demonstrable in 36 of 83 patients. Residual B-cell function was most frequent in the patients with the shortest duration of diabetes. The metabolic importance of endogenous insulin was demonstrated by the significantly lower insulin requirement in the patients with residual B-cell function.


Control of tryptophan synthetase amplified by varying the numbers of composite plasmids in Escherichia coli cells. Using pSC101, RSF1010, RSF2124 and RP4 plasmids as vectors and bacteriophage lambdatrpD-A60-3 DNA as a source of the Escherichia coli whole tryptophan operon, composite plasmids of pSC101-trp, RSF1010-trp, RSF2124-trp and RP4-trp were constructed in vitro with EcoRI restriction endonuclease and DNA ligase. Each composite plasmid could be maintained stably in E. coli cells. The copy number of pSC101-trp, RSF1010-trp, RSF2124-trp and RP4-trp were 4.2, 11.2, 11.9 and 1.6 per chromosome respectively. The tryptophan synthetase activities in cells containing pSC101-trp, RSF1010-trp, RSF2124-trp aand RP4-trp plasmid were found to be 2.1, 6.0, 5.0 and 2.5 times compared with the level in chromosomal trp+ cells when they were grown in a minimal medium. By partial derepression with indolylacrylic acid, the enzyme levels were elevated to 10.1, 16.3, 15.3, 12.3 times, respectively, that of the control cells. The tryptophan synthetase activities did not increase in proportion to the copy number of the plasmids, but were strongly affected by the repression system of host cells.


Sequence organization and expression of a yeast plasmid DNA. Saccharomyces cerevisiae strain A364A D5 contains circular double-stranded DNA molecules of 6230 +/- 30 base pairs (2mu DNA) which are present in 68 copies per cell and make up 2.4% of the haploid genome. About 0.4% of non-poly A containing yeast RNA hybridizes to the yeast DNA circles. When denatured and then self-annealed, the DNA molecules assume a characteristic "dumbbell" shape in the electron microscope indicating that each circle possesses a non-tandem inverted repeat sequence of 630 +/- 10 base pairs. Eco-RI digestion of purified 2mu DNA yields 4 fragments on an agarose gel whose combined molecular mass is twice that of the monomer circle, suggesting that there are 2 populations of circles, each of the same molecular weight. Representatives of each population have been separated by cloning in Escherichia coli via the bacterial plasmid pSC101. Heteroduplex analysis of the cloned circles show that the 2 different populations arise because of intramolecular recombination between the inverted repeat sequences. Acrylamide gel patterns of polypeptides synthesized in bacterial mini-cells containing the hybrid plasmids between 2mu DNA and pSC101 are significantly different than the pattern obtained from mini-cells containing pSC101 alone.


An improved bacteriophage lambda vector: construction of model recombinants coding for kanamycin resistance. An attenuated bacteriophage lambda has been prepared for proposed use as an EK2 vector. This phage, designated lambdagt vir Jam27 Zam718-lambdaB' can accomodate up to 11.10(6) daltons of foreign DNA inserted through Eco RI ends. The virulence mutations and nin 5 reduce the frequency of lysogen and/or plasmid formation. The mutations Jam27 and Zam718 require a suppressor in the bacterial host. The phage recombination functions contained in the EcoRIlambdaC fragment have been deleted, and only the EcoRIlambdaB fragment remains (in reverse orientation) in the center portion of the vector. In addition, this phage adsorbs to sensitive bacteria at a significantly reduced rate, conferring another block to the escape of free phage. Model recombinants have been constructed by in vitro recombination with an EcoRI fragment coding for kanamycin resistance (originally derived from R-factor R6-5). This fragment of DNA is 4.6.10(6) daltons in size, contains an inverted repeat, and also appears to contain a promoter for the kanamycin resistance gene. Using this model recombinant, the rate of transfer of kanamycin resistance to permissive and nonpermissive strains of E. coli has been measured.


Cloning and amplification of alpha and beta mouse globin gene sequences synthesised in vitro. New chimeric Escherichia coli plasmids containing alpha or beta globin gene sequences of the mouse were constructed. Double-stranded DNA, synthesised in vitro in a 2-step reaction from mouse globin mRNA was inserted into E. coli plasmid pCR1, after tailing of the 2 DNAs with dG and dC respectively. Some of the mouse globin plasmids described contain at least 90% of the globin mRNA sequence and therefore contain the entire translated sequence of the globin genes. Some possible uses of these recombinant plasmids are described.


In vitro packaging of a lambda Dam vector containing EcoRI DNA fragments of Escherichia coli and phage P1. In this report we describe a coliphage lambda vector system for cloning endo R. EcoRI DNA fragments. This system differs significantly from those previously described in two ways. First, restricted and ligated DNA is encapsidated in vitro. Second, with increasing lambda DNA size in the range 78 to 100% that of wild-type, the efficiency of DNA encapsidation into infectious phage particles markedly increases. For lambda wild-type DNA the efficiency of in vitro packaging (10(6) to 10(7) plaques produced per microgram of added DNA) is equal to, or better than, the standard CaCl2 transfection method. The use of a Dam mutation to facilitate recognition of size classes of inserted fragments is described. Using this vector and in vitro packaging, several E. coli and phage P1 and R.EcoRI fragments were cloned.


Cloning of chemically synthesized lactose operators. Recombinant DNA molecules, constructed from the ColE1-Mk5 hybrid plasmid PMB9 and a chemically synthesized wild-type lactose operator segment, have been used to transform Escherichia coli. Up to 10% of the transformants (selected for the tetracycline-resistance property of PMB9) are partially constitutive for the lactose operon enzyme beta-galactosidase. In vitro studies demonstrate that these partially constitutive transformants contain plasmid DNA molecules which carry one or more lactose operators, and which will bind purified lactose repressor. Preliminary results with some modified operator sequences are also presented.


Isolation and characterization of the biotin genes of Escherichia coli K-12. DNA containing the biotin gene cluster, bioABFCD, of E. coli K-12 has been isolated from the EcoRI cleavage products of lambdabiot124-10 phage DNA and subsequently characterized by electron microscopic studies. The biotin-DNA fragment obtained after EcoRI cleavage of the lambdabiot124-10 DNA measures 18.7% lambda DNA length (approx. 9000 base pairs). In addition to the biotin genes, it contains 4.75% and 3.08% lambda phage DNA at the left and right end-points of the bioABFCD cluster, respectively. The two bio promoter sites of the divergently transcribed biotin genes have been visualized under the electron microscope by binding RNA polymerase holoenzyme to the biotin DNA fragment. The two promoters are located at 41% and 43% length of the DNA fragment from its left endpoint. In vitro transcription of RNA from the bio-tin-DNA fragment has been visualized with the electron microscope, but so far no simultaneously transcribing "RNA:DNA" loops of the divergently oriented genes have been observed.


Carcinoembryonic antigen (CEA) in the normal human small intestine: a light and electron microscopic study. An immunoperoxidase method for the demonstration of carcinoembryonic antigen (CEA) in tissues was applied to formalin-fixed paraffin embedded and glutaraldehyde-fixed resin sections of normal human small intestine. CEA could easily be demonstrated coating the surface of the small intestine, lining the crypts, and in goblet cells, indicating its presence there in considerable concentration. At the ultrastructural level CEA was localised in the glycocalyx and in mucin granules of goblet cells but not intracytoplasmically.