Simplified method for determination of protocollagen proline hydroxylase. Preparation of (14C) Pro labeled protocollagen and requirements for its hydroxylation by PPH was investigated. Protocollagen can be prepared by centrifugation of the biological material at 15.000 X g. Substances in current use, viz catalase, bovine serum albumin and dithiothreitol were found unnecessary under the hydroxylation conditions used. A decrease in PPH in the testis and skin of rats with increasing age was demonstrated.


Evaluation of a simple cortisol assay by competitive protein binding technique. The evaluation of a recently introduced kit (Cortipac (R)) for measuring plasma cortisol by competitive protein binding technique without prior extraction of steroids from plasma, is presented. The blank value was less than 2.5 mug/100 ml, and the lowest measurable value was 2.5 mug/100 ml. The coefficients of variation within- and between-batch determinations were 8.64% and 9.21% respectively. By adding known amounts of cortisol to pre-extracted plasma a linear correlation between calculated and measured cortisol was obtained. In comparison with the double isotope derivate technique, a linear correlation was found. A high degree of cross-reaction was found with nearly all of 11 tested steroids. Due to the considerably higher plasma cortisol concentration in most clinical cond-tions, the lack of specificity may be of minor importance, but must be considered in evaluating increased plasma cortisol values. The mean plasma cortisol concentration in normal subjects (20.3 mug/100 ml) was equal to or slightly higher than found by other methods. The kit was found appropriate for routine clinical application, due to the easy, rapid and inespensive performance with an acceptable recovery and reliability, but it cannot entirely replace more specific methods.


The use of NADH as a standard in a modified PMS-INT colorimetric assay of lactate dehydrogenase. A new and more reliable way of quantitating lactate dehydrogenase activity, using NADH as a standard in a colorimetric assay, is proposed. Adopting the scheme, we obtained a CV of 2.6% and precision (95% limits) of +/-5.4% for enzyme activities ranging from 110 to 600 U/l. Comparative study against the modified kinetic method of Amador et al. showed excellent correlation, r = 0.99, p  less than 0.001. A normal range of 75 to 203 U/l was determined for the proposed scheme.


Sclerosteosis - an autosomal recessive disorder. Sclerosteosis is a rare, potentially lethal skeletal disorder in which massive bony over-growth leads to facial distortion, cranial nerve compression and progressive rise in intracranial pressure. Gigantism and syndactyly of the 2nd and 3rd fingers are associated features. In a nationwide investigation in South Africa, 25 affected individuals in 15 Afrikaner kindreds have been studied. The minimum prevalence of the contition in this community is 1 in 75,000. Analysis of pedigree data confirms that sclerosteosis is an autosomal recessive condition. The gene frequency in the Afrikaner people is estimated at 0.0035, with 10,000 clinically normal heterozygotes in this population. Heterozygote detection may be possible on a basis of recognition of minor changes which are apparent on skull radiographs.


Transmission of auto-immune haemolytic anaemia and murine leukaemia virus in F1 (BALB/c X NZB) hybrid mice derived by ovum transplantation. F1(BALB/c X NZB)hybrid progeny derived by ovum transplantation were used to study the transmission of auto-immune haemolytic anaemia and murine leukaemia virus (MuLV) by male New Zealand black (NZB) mice. Fertilized ova, collected from the normal BALB/c partners 3 1/2 days after mating, were transferred to other, surrogate, BALB/c mothers, which then carried, delivered, and reared the hybrid young. This technical manoeuvre effectively closed the congenital transplacental route theoretically available to any infectious MuLV originating from the NZB father. Nevertheless, such progeny developed exactly the same profile of auto-immune haemolytic disease and the same range of diverse malignancies as their normally-derived F1(BALB/c X NZB) counterparts, and they carried type C MuLV particles readily detectable by electronmicroscopy. We concluded, therefore, that both the auto-immunity and virus were transmitted before placentation, presumably by the NZB male at fertilization, and probably as genetic information.


The xeroradiographic appearances of some uncommon malignant mammary neoplasms. The clinical and xeroradiographic appearances are described of four patients with malignant lymphomatous involvement of the breast, two with breast involvement by acute leukaemia, and four with primary fibrosarcoma of the breast. In one patient with malignant lymhoma and multiple palpable masses, the radiological appearance resembled benign mammary dysplasia, but in the three other patients, one with a discrete clinical mass, and two with diffuse clinical involvement, radiological examination of the breast showed skin thickening with diffuse abnormalities of the trabecular pattern, most marked in the subdermal area. One of the patients with leukaemia had a discrete palpable mass and the other had diffuse clinical involvement, but the radiological appearances in both patients were similar and resembled those of a carcinoma. The clinical and radiological appearance of the patients with fibrosarcoma were those of a benign cyst or a fibroadenoma.


The scintigraphic demonstration of acute myocardial infarcts. The feasibility of acute infarct scintigraphy for the clinical evaluation of patients with known or suspected acute myocardial infarction is established. Further development of this methodologic approach may result in even better agents for the visualization of infarcts. Radiotracers with high affinity for the infarct, rapid blood clearance, and low concentrations in surrounding organs, such as liver and bone, would be more suitable than available radiopharmaceuticals for acute myocardial infarct scintigraphy. Ultimately, labeling these tracers and ultra-short-lived radionuclides will enable rapid sequential imaging to assess changes in the extent of infarction and to determine the efficacy of therapies aimed at limiting infarct size.


On a shoestring: a consumer-based source of personpower for mental health education. This paper outlines a program for promoting mental health in the population by "giving away our skills" as mental health professionals. This kind of program can be started on a shoestring by a community mental health center, family service agency, or adult education program. It consists of teaching parent-child communication courses, with selected parents then becoming instructors to build a network of nonprofessional personpower. The paper explores strategies of setting up programs. It explores the shifting role of the professional toward a colleague relationship with nonprofessionals. Finally, it presents research on the program's effectiveness with parents and the impact on the lives and self-images of the nonprofessional instructors themselves.


Is NIMH's dream coming true? Wyoming centers reduce state hospital admissions. Do comprehensive community mental health services reduce admissions to state hospital? A recent Nader report says no. Surprisingly few studies have documented the success of comprehensive community mental health programs. Wyoming had four unique characteristics that aided in evaluating the impact of centers on hospital admissions. The results of this study indicated that community-based comprehensive mental health  services do, in fact, reduce psychiatric hospitalization.


The action of arterial hypertension on lipid and lipoprotein metabolism. II. Qualitative and quantitative alterations of blood serum, liver and aortic lipids and lipoproteins in Okamoto-Aoki rats with spontaneous hypertension. The purpose of the study was to investigate alterations in the content of the basic lipid fractions, and of the low density lipoproteins (LDL) in the liver, the blood serum and aorta, as well as to determine by acrylamide disc electrophoresis the hyperlipoproteinaemic type of spontaneously hypertensive rats (SHR) which are considered as the most suitable model of essential hypertension. The experiments were carried out on 25 normotensive control Wistar rats and 30 SHR (Okamoto-Aoki strain). An augmentation of lipid metabolism in the liver and a moderate hyperlipidaemia mainly due to an increase in triglycerides was found. The quantitative alterations of the lipid fractions corresponded with the qualitative alterations of the lipoproteins, an intensive and permanent pre-beta-LP fraction being established. In all the SHR a peculiar pattern of hyperlipoproteinaemia differing from the basic Fredickson-Lees patterns by a LP-fraction located between tha alpha- and beta-LP fractions was also established. The alterations in the lipid and lipoprotein metabolism in SHR are considered as connected with the hypertensive state itself since no accompanying atherosclerosis was observed.


Some aspects of rat kidney plasma membrane cAMP-receptor and its connection with protein kinase activity. Rat kidney plasma membranes prepared by the method of FITZPATRICK et al. (J Biol. Chem. 244, 3561, 1969) show protein kinase activity as well as specific cAMP binding activity (diss. const. 1.3 x 10-9 M). However, no stimulation of kinase activity by cAMP is observed in presence of exogenous substrates (e.g. histone) and only poor stimulation with endogenous substrates in the membrane could be shown. At high ionic strength (1 M NaCl) cAMP independent protein kinase activity can be solubilized. Low ionic strength buffer (1 mM Tris-HCl pH 7.4 1 mM EDTA) and non-ionic detergents (Lubrol PX, Lubrol WX and Triton X 100) are able to solubilize both protein kinase activity and cAMP binding activity. Protein kinase activity seemed to be only loosely associated with the membrane, whereas cAMP binding protein appears to be more firmly fixed into the membrane structure. In addition we have found that membranes serve as a good substrate for cytosol protein kinase (s) and Ca-ion concentration influences the effect of cAMP on protein kinase activity. Dependent on the increase of Ca-ion concentration the effect of cAMP on protein kinase changes from activation to inhibition.


Effect of cyclic nucleotides on the isotonic fluid reabsorption in the proximal convoluted tubule of rat kidney. In order to reveal the sidedness of the cyclic AMP action on renal tubules the effect of cyclic nucleotides on isotonic fluid reabsorption was examined in proximal convoluted rat kidney tubules by using advanced micropuncture techniques. Cyclic AMP inhibited isotonic fluid reabsorption preferentially when applied on the luminal cell side. This finding coincides with a predominant distribution of a membrane bound cyclic AMP dependent protein kinase in a luminal cell membrane fraction. On the luminal cell side cyclic AMP and dibutyryl cyclic AMP inhibited isotonic fluid reabsorption in a similar dose dependent behaviour. Concentrations of both nucleotides as low as 10-10M had a definite inhibitory effect. Tested at a high concentration N6-butyryl cyclic AMP was almost as effective as cyclic AMP. Deoxy cyclic AMP, 5' AMP, cyclic GMP and dibutyryl cyclic GMP had no effect. ATP inhibited isotonic fluid reabsorption to a very small extent. The data indicate that the observed inhibition of isotonic fluid reabsorption is specific for cyclic AMP and that the interaction of cyclic AMP and the luminal cell membrane is initiated at the luminal cell surface.


Role of mitochondrial Ca2+ in antidiuretic hormone action. The calcium ion concentration measured in rat kidney mitochondria, isolated from vasopressin treated tissue, has a dose response characteristic in which the calcium concentration reached a minimum at low doses of vasopressin (2 mU/ml), at higher doses of hormone the mitochondrial calcium ion concentration increases reaching a value close to that of the controls with vasopressin (100 mU/ml). This efflux and subsequent uptake of mitochondrial calcium has been shown to be a direct effect of the varying cyclic AMP concentrations. Sodium and water permeability effects of vasopressin have been shown in toad bladder to have different dose response characteristics. Maximum sodium transport occurs at a lower dose of vasopressin (2 mU/ml) and is believed to be associated with direct permeability effects of the hormone. Maximum water transport occurs at a higher dose of vasopressin (100 mU/ml) over a concentration range associated with hormone-stimulated adenylate cyclase activity. The water transport response to low doses of vasopressin may be potentiated by aldosterone treatment, an effect that can be related to the inhibition of tissue phosphodiesterase activity and subsequent increased cyclic AMP concentrations. In steroid depleted conditions the cyclic AMP medicate efflux of mitochondrial calcium ions, that occurs at low doses of vasopressin, may prevent the release of membrane bound calcium ions and thus inhibit the water permeability effect of the hormone. Higher levels of cyclic AMP reverse this inhibitory effect and give rise to an increased water flow. It is concluded that cyclic AMP and intracellular concentrations of calcium ion act as inter-related mediators of antidiuretic hormone action.


Effect of an ammonia load on the kidney near-equilibrium systems in the rat in vivo. 1)The time course of changes in concentration of renal metabolites in response to a non-toxic load of NH4 as NH4 Cl or NH4HCO3 were measured in fasted rats. 2) Following a NH4Cl load, decrease of renal concentration of 2-oxoglutarate occurs but this change is delayed in relation to the peak of the blood ammonia concentration and persists after disappearance of the hyperammoniemia. 3) Following a NH4HCO3 load, the oxoglutarate concentration changes are less marked and more transient. 4) No close relationship between the mitochondrial free NAD/NADH ratio calculated from the glutamate dehydrogenase and the 3-hydroxybutyrate dehydrogenase systems were seen during alteration of the ammonia concentration. 5) Contrary to the observations in the liver under similar circumstances (BROSNAN, J.T. et al.: Biochem.J. 138, 453, 1974), no increase in kidney tissue or renal venous blood alanine or aspartate concentration are seen. 6) A constant infusion of NH4HCO3 resulted only in an increase in tissue and renal venous blood glutamine concentration. 7) The infusion of NH4 together with a carbon source (malate) resulted in a similar increase in tissue glutamine concentration and more striking increase in renal venous glutamine concentration. No accumulation of aspartate nor alanine were seen. 8) In vitro studies indicate that the net flux through both the aspartate aminotransferase and the glutamate dehydrogenase reactions is dependent on the concentration of the reactants as expected for a near-equilibrium system. 9) It is concluded that the kidney response to an ammonia load differs from that of the liver despite the existence of a similar network of near-equilibrium reactions of (1) a lack of local availability of oxaloacetate, (2) a lower activity of alanine aminotransferase, (3) a greater in vivo activity of glutamine synthetase.


Regulation of phosphoenolpyruvate carboxykinase by glutamine and ATP as possible control mechanisms of renal gluconeogenesis. In different metabolic states renal phosphoenolpyruvate carboxykinase (PEP-CK) activities are closely correlated with in vitro glucogenic rates, suggesting a limitation of the glucogenic capacity of kidney by this enzyme. Stimulation of renal gluconeogenesis from pyruvate, lactate, and succinate by lysine and glutamine was therefore associated with a regulatory attack of these amino acids at the level of PEP-carboxykinase. This postulate was confirmed by the failure of lysine to stimulate glucose synthesis from fructose. Experimental support for an interference of glutamine and PEP-carboxykinase was obtained by a study on the inactivation of this enzyme in kidney cortex homogenates: A rapid inactivation of enzyme activity within 40-50 min could be slowed down by glutamine. In addition the inactivation was counteracted by ATP. At suboptimal concentrations of the trinucleotide its effect was potentiated by c-AMP and c-GMP. Studies on the effect of ATP on PEP-carboxykinase in kidney cortex homogenates from rats in different metabolic states revealed: In homogenates from carbohydrate fed animals extreme low activities of PEP-CK were not altered by ATP, whereas elevated enzyme activities after a protein rich diet could be further raised by a factor of 2 or 3 by ATP. GTP and ITP could substitute for ATP. An extension of these studies on hepatic enzymes showed a similar inactivation of tyrosine aminotransferase (TAT) and a protective effect of ATP. The data obtained from these experiments favour an interconversion of PEP-carboxykinase and tyrosine aminotransferase into different forms as possible mechanism for their regulation.


The mode of activation and regulation of glutaminase in intact kidney mitochondria. Experiments were carried out with isolated pig renal-cortex and rat kidney mitochondria. Respiration, glutaminase activity and swelling of the mitochondria were followed by an oxygen electrode, an NH+4-sensitive electrode and spectrophotometrically. The inhibitors of the transport of glutamate (avenaciolide) and phosphate (mersalyl) across the inner mitochondrial membrane were employed. On the basis of the experimental findings we made the following conclusions: 1. There are two mechanisms by which glutamate can leave the inner space of kidney mitochondria: (a) an electrogenic efflus coupled to the respiration driven proton translocation and the presence of a membrane potential (positive outside), and (b) an electroneutral glutamate-hydroxyl exchange which is inhibited by avenaciolide and which operates in both directions. 2. The activation of glutaminase in intact mitochondria by phosphate occurs only if the activator moves across the inner mitochondrial membrane. The enzyme can be activated even when the movement of phosphate is inhibited provided that the mitochondria are energized. 3. Glutaminase is intimely associated with the inner mitochondrial membrane so that the part of the enzyme with the binding site for phosphate is embedded in the membrane, whereas the part with the binding sites for the substrate and glutamate is exposed toward the matrix space. The mechanism of allosteric activation of glutaminase and of the efflux of glutamate from mitochondria are probably the crucial factors which regulate the enzyme activity, and, consequently, renal ammoniagenesis in vivo.


Mitochondrial abnormalities in progressive ophthalmoplegia. A clinically diagnosed case of progressive external ophthalmoplegia was biopsied and ultrastructurally evaluated. The disease was found to be characterized by numerous giant mitochondria. The intramitochondrial changes consisted of either swirls of cristae or of paracrystalline structures which were in apposition with the membranes of the cristae. The internal structure of the paracrystalline units consisted of parallel bands of material which were aligned in the same plane as the membranes of the related cristae.


Electron microscopy of the cotton fibre: new observations on cell wall formation. The ultrastructure of the cotton fibres was examined after developing successful fixation methods. Fibre cells were fixed at different stages of development. In cells which were elongating and producing primary cell walls, the Golgi apparatus appeared to be directly involved in secretion and synthesis of primary wall components. In cells which were synthesizing thick secondary cell walls, evidence suggested a major role for the endoplasmic reticulum and plasma memebrane in the synthesis and secretion of secondary wall materials. The possibility of a shift from a Golgi apparatus pathway for primary wall synthesis to an endoplasmic reticulum pathway for secondary wall synthesis is discussed. Plasma membrane micro-invaginations are present only during secondary wall synthesis and may represent sites of cellulose assembly. A model for primary wall biogenesis via the Golgi apparatus is presented, and the potential of the cotton fibre as a model system for studying cellulose biogenesis in higher plants is discussed.


Plastoglobules of leaf chloroplasts of two cultivars of Capsicum annuum. The chloroplasts of the leaves of Capsicum annuum cv. Purple Leaf Cherry are characterized by the presence of large electron translucent plastoglobules which comprise a considerable proportion of the plastid volume. The leaf chloroplasts of Purple Leaf Yellow cultivar contain, in addition, large starch granules which are frequently in intimate association with these plastoglobules. There appears to be a controlled expulsion of plastoglobules into the vacuole by the non-degenerating chloroplasts of both cultivars, although the function of this secretion is not known. The chloroplasts of older leaves of Purple Leaf Yellow also contain plastoglobules with a nonhomogenous fine structure. Some have granular segments, membrane-like bands, or myelin-like bands, the latter suggesting the presence of glyco- and/or phospholipids in such plastoglobules.


Bronchilar carcinoma in a 20-year-old man. A 20-year-old man with a nine-month history of pulmonary abscess and hemoptysis was discovered to have a small nidus of bronchiolar carcinoma in the fibrotic reaction surrounding the pulmonary abscess. The cell type, origin, and occurrence of this tumor in fibrotic pulmonary disease are discussed. We believe that this case represents an unusual presentation of bronchiolar carcinoma with respect to clinical manifestation, tumor size, and patient's age.


Chronic eosinophilic pneumonia. Ultrastructural evidence of marked immunoglobulin production plus macrophagic ingestion of eosinophils and eosinophilic lysosomes leading to intracytoplasmic Charcot-Leyden crystals. A 60-year-old man was diagnosed as having chronic eosinophilic pneumonia by clinical presentation and open lung biopsy. Electron microscopic study of the tissue from biopsy revealed evidence of marked production of immunoglobulin, which may be part of the chemotactic process which attracted large numbers of eosinophils into the area. Also, there were macrophages which were phagocytosing degenerating eosinophils and eosinophilic lysosomes. Charcot-Leyden crystals were noted in the cytoplasm of many macrophages. The relationship of the production of immunoglobulin to the eosinophilia and the mechanism of intramacrophagic formation of Charcot-Leyden crystals is discussed.


Discharge planning in foster care cases where the father is the significant parent. In planning for the discharge from foster care of children in one-parent families, the potential of fathers is often neglected. An agency study indicates that intensive casework and adequate services can make the return to fathers appropriate.


[Nasal application of a synthetic alpha1-18 corticotropin: an effective form of ACTH treatment (author's transl)]. The absorption of a synthetic ACTH-peptide (alpha1-18 corticotropin, Ba 41.795) via the nasal mucosa was investigated in twelve probands. After application of 1 mg into each nostril a rapid increase of the plasma cortisol was observed which returned to the initial value only after 7 hours. In two patients with rheumatoid arthritis and two patients with bronchial asthma who had been treated with regular steroid injections an attempt was made to replace the injections by nasal application. The same therapeutic effects could be obtained, but larger doses were required than when using injection therapy.


Regulation of lactogenic hormone binding in rat liver by steroid hormones. We have measured the effects of testosterone propionate, medroxy-progesterone acetate and cortisol on the binding of ovine [125I]iodoprolactin to 100,000 X g particulate fractions from liver of normal and estrogen-treated female rats. In untreated animals 6.7 +/- 1.1% (SD) of the radioactivity added to 0.5 mg of membrane protein was specifically bound to the hormone receptor. Specific binding was significantly (P less than .05) decreased after 7 daily doses of testosterone (1.0 mg) to 2.8 +/- 1.4%, medroxyprogesterone (0.25 mg) to 2.7 +/- 0.2% and cortisol (5.0 mg) to 3.1 +/- 1.3%. The serum prolactin concentration, 4.2 +/- 3.4 ng/ml in normal animals, was not affected by the hormone treatment. Ethinyl estradiol, 10 mug/day for 7 days, increasing the binding of [125I]iodo-prolactin to 16.6 +/- 6.0% and increased serum prolactin to 50.6 +/- 11.5 ng/ml. Simultaneous administration of testosterone, medroxyprogesterone or cortisol with estradiol did not diminish the estradiol-induced increase in serum prolactin, but completely prevented the increase in prolactin binding. Testosterone or cortisol given to animals pretreated with estradiol suppressed prolactin binding from 16.4 +/- 4.2% to less than 2.5% after 48-72 h. Parellel results were obtained with 125I labeled human growth hormone whereas 125I labeled-insulin binding was not affected by these treatments. Scatchard analysis showed that the decrease in lactogenic hormone binding was due to a reduced concentration of receptors with no significant change in affinity. Since serum levels of prolactin were not changed, we conclude that treatment with testosterone, medroxyprogesterone, and cortisol decreased lactogenic hormone binding by a direct action on the liver.


Circadian patterns of stress-induced ACTH secretion are modified by corticosterone responses. To test whether there is a circadian rhythm in the ACTH response to stress, young female rats were exposed to a variety of ACTH-releasing stimuli at 0600 and 1800 h and changes in circulating ACTH and corticosterone were measured. The results of these experiments suggested that after the high intensity stimuli of laparotomy with intestinal traction or 250 mug histamine ip/100 g BW, the morning ACTH response was greater than the evening response. However, the ACTH response to ip saline was greater in the evening in one experiment and greater in the morning in a second experiment. Plasma corticosterone responses were faster and greater in the morning in the first experiment and in the evening in the second experiment. The ACTH response to 125 mug histamine ip/100 g BW was greater in the evening and the change in corticosterone was greater in the morning. Thus, after low intensity stimuli, the ACTH responses appeared to depend on both the lag time prior to the corticosterone response, and its magnitude. To test this possibility, rats were adrenalectomized and the ACTH response was measured 7.5 and 15 min after the start of surgery and after injection with either 2% EtOH-saline, or 50 mug corticosterone at operation, or with 30 mug corticosterone at 5 min. Compared with ACTH levels in rats treated with vehicle, ACTH levels were decreased 7.5 min after 50 mug corticosterone at operation (P less than 0.01), but not after 30 mug corticosterone at 5 min. ACTH levels were slightly reduced 10 min after 30 mug corticosterone at 5 min compared with those of rats injected with vehicle at operation (P less than 0.05). These results are consistent with the interpretation that corticosterone secretion modifies stress-induced ACTH secretion via the fast-feedback effect. Comparison of the ACTH responses to acute adrenalectomy plus injection with EtOH-saline at 0600 and 1800 h demonstrated that, in the absence of a corticosterone response to the stress, the ACTH response is greater in the morning that in the evening (P less than 0.01). Finally, this group of experiments suggests strongly that the responsivenss of rat adrenal glands to ACTH increases markedly between 0600 and 1800 h.


Interaction of human growth hormone with isolated rat liver cells. The interaction of 125I-labeled human growth hormone (hGH) with isolated rat liver cells is a specific, time dependent and saturable process. In male rats, one cell binds a maximum of 2000 hormone molecules; the dissociation constant of the cell-hGH interaction is about 3 X 10(-10)M. Liver cells of female rats bind 5 to 10 times more hGH than do those of male rats at equivalent hormone concentrations. Binding of 125I-labeled hGH to liver cells is readily inhibited by native hGH; 50% inhibition occurs at about 2 X 10(-9)M hGH irrespective of sex. In male rats, bovine growth hormone (bGH) is almost as potent as hGH in inhibiting 125I-labeled hGH binding; no displacement occurs with ovine prolactin (oPRL) except at very high (greater than 10(-6)M) concentrations. In female rats, bGH competes less effectively, and oPRL, more effectively, than they do in males; in addition, oPRL demonstrates a higher apparent affinity for the hGH binding sites (4 X 10(-9)M) than does bGH (1 X 10(-8M). These findings suggest that in female rats hGH, unlike bGH, interacts with additional, "lactogenic" binding sites that are distinct from the "growth hormone" binding sites. The 125I-labeled hGH eluted from liver cells as well as that which remains in the incubation medium retains full biological activity, as judged on its ability to bind specifically to liver membranes. Treatment of liver cells by phospholipase A causes a 5-fold increase in cell binding capacity. Liver cells bind about the same amount of hGH as do crude particulate fractions from these cells; this suggests that in the intact cell, binding occurs at relatively accessible sites, presumably localized in the plasma membrane.


Stimulation in vivo of the secretion of prolactin and growth hormone by beta-endorphin. Morphine sulfate (MS) and the opioid peptide beta-endorphin beta-LPH-(61-91) stimulate prolactin and growth hormone release in steroid-primed and non-treated male rats when injected intravenously or intracisternally. On a molar basis beta-endorphin is at least 20 times more potent than MS, whereas Met5-enkephalin (beta-LPH-(61-65)) and alpha-endorphin (beta-LPH-(61-76)) are devoid of activity at the dose injected (300 mug). The in vivo stimulatory effects of beta-endorphin on prolactin secretion are reversed by the opiate antagonist naloxone. The absence of in vitro effect of MS and beta-endorphin on prolactin and growth hormone secretion by cultured rat pituitary cells suggest that they have a central nervous system site of action.


(Des-Asp1) angiotensin I: a study of its pressor and steroidogenic activities in conscious rats. The steroidogenic and pressor activities of the nonapeptide (des-Asp1) angiotensin I [(des-Asp)-AI] were tested in conscious rats. (des-Asp)-AI caused dose related increases in mean arterial pressure (MAP), serum aldosterone, and serum corticosterone in doses between 3 and 3,000 ng/kg/min. (des-Asp)-AI was 14% as potent as angiotensin I and angiotensin II and 60% as potent as (des-Asp1) angiotensin II [des-Asp)-AII] in raising MAP. (des-Asp)-AI was less active than AI, AII, or (des-Asp)-AII in causing increased release of aldosterone, possessing only 8%, 11%, and 17% of the potency of AII, (des-Asp)-AII, and AI, respectively. Each of these angiotensin peptides was equally potent in elevating serum corticosterone levels. Infusions of a nonapeptide inhibitor of converting enzyme (CEI, 0.5 mg/kg/min iv) did not alter control MAP or blood pressure responses to AII or (des-Asp-)-AII but inhibited equally the blood pressure effects of AI and (des-Asp)-AI. CEI also inhibited the ability of (des-Asp)-AI (67% inhibition) and AI (34% inhibition) to increase the serum aldosterone concentration, but had no effect on basal aldosterone levels. These data indicate that (des-Asp)-AI has pressor and steroidogenic effects, but requires conversion to (des-Asp)-AII for a major portion of its activity. These results further substantiate the hypothesis that (des-Asp)-AII, recently recognized as a hormone of the renin-angiotensin system, may be produced without the formation of AII as an intermediate and provide in vivo evidence for the conversion of (des-Asp)-AI to (des-Asp)-AII.


Evidence for the existence of a histamine H2-receptor in the mouse thyroid. The existence of a histamine H2-receptor in the thyroid was investigated. Histamine in vitro stimulated the formation of cyclic AMP and colloid droplet formation in mouse thyroid lobes. Stimulation by histamine of cyclic AMP formation in mouse thyroid lobes was significantly inhibited by metiamide, a histamine H2-receptor antagonist. 4-Methylhistamine, a histamine H2-receptor agonist, markedly stimulated cyclic AMP formation, whereas 2-methylhistamine, a histamine H1-receptor agonist, was ineffective. The stimulation by 4-methylhistamine of cyclic AMP formation was markedly inhibited by metiamide, but not by chlorpheniramine, a histamine H1-receptor antagonist. In contrast, metiamide did not affect cyclic AMP formation induced either by TSH or by the long-acting thyroid stimulator. Therefore, it is suggested that there exists a histamine H2-receptor in the membranes of the thyroid follicular cells which facilitate thyroid hormone secretion via the adenylate cyclase-cyclic AMP system.


Further studies on the mechanisms controlling prostaglandin biosynthesis in the cat adrenal cortex: the role of calcium and cyclic AMP. In light of previous studies which have implicated prostaglandin (PG) formation as a link in ACTH-induced steroid production by isolated cat adrenocortical cells, experiments were carried out to provide additional information regarding the role of PGs in adrenal steroidogenesis and their interactions with calcium and cyclic AMP. Perfusion of cat adrenal glands with Locke's solution plus beta(1-24)-ACTH resulted in an immediate increase in PGF2alpha release, which rapidly declined to basal levels after the stimulus was withdrawn. In contrast, maximal rates of steroid release were manifest some 30 min after removal of ACTH. ACTH and its onitrophenyl sulfenyl derivative (NPS-ACTH) increased PG (PGF2alpha and PGE2) and steroid release by trypsin-dispersed cat cortical cells, but NPS-ACTH, unlike ACTH, did not augment cortical cyclic AMP levels. In this same preparation, indomethacin completely blocked ACTH and NPS-ACTH facilitated PGF2alpha and PGE2 release but failed to suppress steroid release markedly. Calcium-deprivation blocked PG and steroid release evoked by these two polypeptides, and depressed PG release elicited by monobutyryl cyclic AMP (bcAMP) without affecting steroid release. These experiments offer additional evidence to support the concept that PGs play a role in the mode of action of ACTH; however, they do not appear to be obligatory intermediates in the steroidogenic process. The importance of calcium in regulating PG formation is discussed with special regard for the idea that this cation has a direct action on the enzyme systems which control PG synthesis.


The inhibitory effect of acute administration of excess iodide on the formation of adenosine 3', 5'-monophosphate induced by thyrotropin in mouse thyroid lobes. In a previous paper, we demonstrated that an inhibitory action of excess iodide on thyrotropin-induced thyroid hormone secretion occurs at a site subsequent to the generation of cyclic AMP. In the present study, however, we have found that thyroidal cyclic AMP formation induced by thyrotropin in vitro was markedly inhibited by the acute administration of excess iodide to mice fed a low iodine diet. In contrast, excess iodide failed to produce inhibition in animals fed a regular diet. In vitro stimulation by long-acting thyroid stimulator (LATS), prostaglandin E2, and 4-methylhistamine of cyclic AMP formation in mouse thyroid lobes was also significantly inhibited by the acute in vivo administration of excess iodide. The inhibition was completely relieved by the administration of methimazole prior to excess iodide. Furthermore, it has been shown that thyroid adenylate cyclase activity induced by thyrotropin was markedly depressed by excess iodide under similar experimental conditions. Therefore, it is suggested that one of the inhibitory actions of excess iodide is on the adenylate cyclase-cyclic AMP system and further, that iodide can elicit its inhibitory action after its conversion to some form of organic iodine.


Effects of follicle stimulating hormone and luteinizing hormone upon cyclic AMP and cyclic GMP levels in rat ovaries in vitro. In vitro effects of FSH and LH on adenosine 3',5'-cyclic monophosphate (cyclic AMP) and guanosine 3',5'-cyclic monophosphate (cyclic GMP) levels in rat ovarian tissue were studied. Ovaries from immature rats treated with either pregnant mare's serum (PMS) or PMS and human chorionic gonadotropin (hCG) were incubated at 37 C for 30 min in defined medium 199 containing aminophylline (0.1 mg/ml). Increasing concentrations of LH (0.4-10.0 mug) when added to the medium containing ovaries with predominantly follicular tissue (PMS-treated) produced progressive increases in cyclic AMP (29-300%) and concomitant decreases in cyclic GMP (41-70%). FSH (4-100 mug) added to the medium produced comparable changes in nucleotide levels: cyclic AMP increased 52-390% and cyclic GMP decreased 45-80%. Ovaries with predominantly luteal tissue (PMS- and hCG-treated) responded differently to the same experimental conditions. LH produced a much greater increase in tissue cyclic AMP levels (209-1280%), whereas FSH produced comparable increases in cyclic AMP as seen with follicular tissue (103-570%). Neither gonadotropin caused any changes in cyclic GMP levels in luteinized ovaries. These results indicate that rat ovarian cyclic GMP is regulated by gonadotropins and the differences in the direction of nucleotide response to gonadotropins suggest divergent roles for cyclic AMP and cyclic GMP in ovarian function.


Ovarian follicular development in the rat: hormone receptor regulation by estradiol, follicle stimulating hormone and luteinizing hormone. The effects of estradiol, FSH and LH on ovarian follicular development and granulosa cell differentiation were examined in the immature rat hypophysectomized on day 24 of age. Administration of estradiol to hypophysectomized rats for 4 days stimulated the growth of large preantral follicles with a concomitant 1.5-fold increase in FSH receptor content and a 4-fold decrease in LH receptor content in the granulosa cells. When highly purified hFSH was administered alone, receptor content for FSH increased progressively for 4 days while receptor for LH remained essentially unchanged. However, when rats were pretreated with estradiol, the response of follicles to FSH was markedly enhanced as indicated by the appearance of large, antral follicles and elevated receptor content for both FSH and LH. Receptor content for FSH increased markedly in response to hFSH following only one day of estradiol pretreatment, while receptor content for LH increased most rapidly in response to hFSH after 3 days of estradiol pretreatment. LH administered to rats possessing large preovulatory follicles caused luteinization of granulosa cells and a marked decline in receptor content for both gonadotropins within 24 h. Receptor content remained low even 48 h after LH administration when granulosa cells were fully luteinized. These results indicated that follicular development and granulosa cell differentiation are dependent on steroid-protein hormone regulation of hormone specific receptors.


Luteal cell receptor content for prolactin (PRL) and luteinizing hormone (LH): regulation by LH and PRL. The effect of LH and PRL during the differentiation of granulosa cells to luteal cells was examined by determining the ability of LH and PRL to regulate luteal cell receptor content for these hormones and to increase production of progesterone. Preovulatory follicles and corpora lutea were hormonally induced in immature hypophysectomized female rats by sequential treatment with estradiol, hFSH and oLH. The content of receptor for LH was high in granulosa cells of large antral follicles. Administration of LH caused receptor for LH to decrease markedly within 24 h and to remain low for 96 h. In contrast, granulosa cell content of receptor for PRL increased progressively for 48 h following LH stimulation and remained elevated in fully luteinized cells at 96 h. This increase in PRL receptor appears to be functionally related to the ability of luteal cells to respond to PRL. When PRL was given for 4 days after LH, both luteal cell progesterone production and LH receptor content increased progressively after, but not before, 48 h. Since these changes occurred in the absence of LH, the increase in LH receptor appears to be a consequence of, but not a requirement for, the PRL-induced increase in progesterone production. If daily injections of PRL were delayed for 72 or 96 h following LH induction of lutenization, luteolytic rather than luteotropic effects of PRL were observed. Since receptor for PRL remained elevated at 72 and 96 h, intracellular mechanisms and not receptor content, appear to be effecting the response of luteal cell to PRL.


Amino acid transport in follicles isolated from the rabbit ovary. The model amino acid alpha-aminoisobutyric acid (AIB) has been used to investigate amino acid transport in follicles isolated from rabbit ovaries. LH markedly increased AIB transport, while incubation with FSH, cAMP or dbcAMP failed to increase the process. Several kinds of inhibitors have been utilized to characterize follicular AIB transport as an energy dependent process and apparently independent of follicular steroidogenesis. The rates of AIB efflux from preloaded follicles were measured in the presence and absence of LH; the rate measurements suggest that only the rate of entry of AIB into follicular cells is stimulated by the gonadotropin. Further experiments used inhibitors of RNA and protein synthesis to examine the relationship between AIB transport and the synthesis of macromolecules. Whereas blocking RNA synthesis suppressed the LH effect on AIB transport, inhibitors of protein synthesis increased the rate of AIB movement into follicles. The in vitro transport of AIB was also studied in follicles isolated 2 hr after mating and in ovulated follicles which were obtained 12 h after coitus. AIB transport rate showed a rise following mating but fell to less than precoital rates in ovulated (12 h) follicles. Transport in both 2 and 12 h follicles proved unresponsive to LH stimulation. Taken together, these studies suggest that AIB transport in isolated ovarian follicles is a LH-sensitive, energy-dependent process. Although AIB transport changes in a pattern qualitatively similar to that previously reported for postcoital protein synthesis, LH stimulation of the two processes may be via different mechanisms.


Dibutyryl cyclic AMP, adenosine and guanosine blockade of the dopamine, ergocryptine and apomorphine inhibition of prolactin release in vitro. Rat anterior hemipituitaries were incubated in Krebs-Ringer bicarbonate containing [3H]leucine. Newly synthesized [3H]prolactin and [3H]GH in the pituitary and incubation medium were assayed, as was the radioimmunoassayable prolactin released into the medium during a 5-h incubation. Dopamine (7.5 X 10(-8)M), ergocryptine (4 X 10(-10) M) and apomorphine (6 X 10(-8)M) all significantly inhibited both radioimmunoassayable prolactin release and newly-labeled [3H]prolactin release without affecting [3H]GH release. Conversely, dibutyryl cyclic AMP (2.5 mM) stimulated radioimmunoassayable prolactin release as well as [3H]prolactin and [3H]GH release. The addition of 2.5 mM dibutyryl cyclic AMP to media containing dopamine, ergocryptine or apomorphine completely restored both radioimmunoassayable prolactin release and [3H]prolactin release to at least control levels. Dopamine, ergocryptine and apomorphine all inhibited incorporation of [3H]leucine into prolactin but not into GH, whereas 2.5 mM dibltyryl cyclic AMP with any one of the inhibitors restored total incorporation into [3H]prolactin to levels insignificantly lower than the nucleotide-stimulated incorporation. Adenosine and guanosine at 2.5 mM also stimulated incorporation into [3H]prolactin and blocked the inhibitory effects of apomorphine upon [3H]prolactin synthesis and release. These nucleosides also stimulated [3H]GH release; and guanosine, but not adenosine, stimulated incorporation into [3H]GH. The ability of dibutryl  cyclic AMP to block the effects of dopamine, ergocryptine and apomorphine upon prolactin release is consistent with these three inhibitors acting by a common mechanism. Cyclic AMP could be hypothesized as a second messenger for prolactin release, but the ability of adenosine and guanosine to mimic almost perfectly the effects of this cyclic nucleotide does not allow any conclusive interpretation.


Strain-dependent gonadal effects upon the response of adrenal cholesterol esters to ACTH in C57BL/10J and DBA/2J mice. The response of adrenal cholesterol esters to ACTH administered in vivo was studied in males, females and male castrates of the C57BL)10J and DBA/21 strains of mice. The cholesterol ester fatty acids were transmethylated and analyzed by gas-liquid chromatography. There are sex and strain differences in the response of the adrenal cholesterol esters to ACTH. Neither the male nor the female C57 mouse shows a significant depletion of total cholesterol esters upon ACTH treatment. This result argues against the utilization of adrenal cholesterol esters for corticosteroid synthesis in this strain. In the female DBA mouse cholesterol ester depletion in response to ACTH is maximal. Neither the total cholesterol ester concentration nor the concentration of any individual ester is correlated with the pattern of response of the cholesterol esters to ACTH. Among the individual cholesterol esters, cholesteryl arachidonate (C20:4) is preferentially utilized after ACTH treatment in all groups except the C57 males. In contrast, cholesteryl adrenate (C22:4), which is generally the most abundant cholesterol ester, is conserved during ACTH stimulation in all groups except the DBA females. A cyclic interconversion between adrenate and arachidonate may be physiologically important in the control of adrenal function.


Somatostatin inhibits the release of acetylcholine induced electrically in the myenteric plexus. Somatostatin inhibits the release of acetylcholine electrically induced in the myenteric plexus-longitudinal muscle of the guinea pig ileum. This effect is not mediated by opiate receptors. Tachyphylaxis to this effect of somatostatin has been observed.


Effect of heparin and adrenocorticotropin on the hepatic and serum cholesterol in catfish, Heteropneustes fossils. The administration of heparin with or without ACTH significantly decreased hepatic cholesterol content in catfish. In serum, heparin alone produced first hypercholesterolemia which was followed by hypocholesterolemia whereas it potentiated hypercholesterolemic action of ACTH three hours after administration. It is concluded that heparin normally caused hypercholesterolemia by releasing cholesterol from liver as lipoprotein complex. The hypocholesterolemic action of heparin might be an indirect one.


Effect of ACTH and its fragment on brain catecholamines in intact and adrenalectomized rats. Brain catecholamine metabolism was monitored by distribution of labelled noradrenaline (3H-NA) after intraventricular injection to intact and adrenalectomized rats. The adrenalectomy produced an increased disappearance rate of the labelled pool in the hypothalamus, hippocampus and neocortex. These changes could be prevented by hydrocortisone pretreatment. Painful stimuli resulted in an increased disappearance of the labelled pool in both intact and adrenalectomized rats. The implantation of hydrocortisone into the tuberoinfundibular region prevented the stress-induced changes of the catecholamine metabolism. Intraventricular administration of ACTH1-24 and ACTH4-10 produced a significant increase of the disappearance rate in different brain regions of adrenalectomized rats. The blocking of catecholamine synthesis by intraventricular injection of alpha-methyl-m-tyrosine resulted in a marked decrease of the labelled pool but did not prevent the ACTH-induced decrease of the tracer pool. On the other hand, the blocking of monoamine-oxydase activity by Pargyline led to a marked increase of the labelled pool but intraventricular administration of ACTH led to an increase of the disappearance rate. The mechanism of ACTH action on brain catecholamine metabolism is still obscure, however, an increased release of the NA to ACTH peptides is very likely in the light of the present observations.


The effect of maternal and fetal corticosteroids on the development of function of the pituitary-adrenocortical system. The pituitary-adrenocortical system of rat fetuses was stimulated (larger adrenals at birth) by maternal adrenalectomy, or suppressed (smaller adrenals at birth) by implantation of an ACTH secreting pituitary tumor (MtTF4). Offspring were delivered by caesarean section and fostered to untreated females. Offspring of intact females delivered by caesarean section and normally delivered offspring of intact mothers both fostered to untreated lactating females served as controls. Body growth in the first three weeks of life was delayed in offspring of tumor bearing mothers in Control-fostered subjects as compared to the 2 other groups. At 70 days of age female offspring of the tumor implanted and adrenalectomized mothers, as well as the Control-caesarean females, had smaller adrenals than Control-fostered animals of the same sex. The adrenal size of males was not significantly affected. No significant differences were found in resting concentrations of corticosterone in plasma, although offspring of adrenalectomized mothers had high values. Suppressed adrenal response to ether stress was found in offspring of tumor bearing mothers. The supposition is that interference with maternal pituitary-adrenocortical activity during pregnancy has a long lasting effect on fetal hypothalamo-pituitary-adrenocortical system.


4-14C-progesterone conversion by the fetal rat adrenal gland in vitro. The adrenal glands of rat fetuses with activated or inhibited pituitary adrenocorticotropic activity between the 15th and 22nd day of intrauterine development were incubated with 4-14C-progesterone for 3hr. Fetuses of intact mothers were used as controls. Conversion of progesterone into adrenal steroids was found increased on the 18th day of intrauterine development, i.e., at the time when fetal adrenocorticotropic activity begins. In comparison to controls, conversion of progesterone into fetal adrenal corticosteroids was the smallest in the fetuses of mothers with inhibited pituitary ACTH and the greatest in the adrenals of fetuses of mothers with activated pituitary adrenocorticotropic activity.


Specific and nonspecific multiple unit activities during the onset of pentylenetetrazol seizures. II. Acute lesions interrupting nonspecific system connections. Nonspecific cortical, thalamic, mesencephalic, and pontine multiple unit activities (MUA) and changes in EEG and MUS of the sciatic nerve after threshold pentylenetetrazol activation were studied in three groups of animals in which neuronal connections were interrupted at three different levels of the central nervous system: spinal, mesencephalic, and prethalamic. Maximal increments of nonspecific MUA and maximal increments and maximal decrements of sciatic MUA after pentylenetetrazol from each group of lesioned animals were statistically compared with tose observed in intact animals. 1. Pentylenetetrazol threshold for producing cortical tonic-clonic EEG discharges was increased in animals with nesencephaic and prethalamic lesions but was not modified in animals with spinal transection. 2. Cortical MUA maximal increment was significantly decreased in mesencephalic and prethalamic lesioned animals, whereas thalamic MUA maximal increment was significantly decreased in mesencephalic and significantly increased in prethalamic lesioned animals. Pontine MUA maximal increment was significantly increased in spinal, mesencephalic, and prethalamic lesioned animals, and mesencephalic M-A maximal increment was not significantly modified in either prethalamic lesioned or in spinal transected animals. 3. Sciatic MUA maximal increment and maximal decrement were significatly decreased in spinal transected animals, whereas only maximal increment was significantly decreased in mesencephalic and only maximal decrement was significantly decreased in prethalamic lesioned animals. These results based on lesion experiments permit us to infer than under normal cinditions the development of generalized seizures induced by threshold pentylenetetrazol injection is highly dependent upon the neuronal interactions between nonspecific structures at different levels of the central nervous system. The possible nature of these neuronal interactions in the intact animals is discussed.


Alpha and beta adrenergic sensitivity in trained and untrained albino rats. The effects on the blood pressure and heart rate responses of different adrenergic stimulants (norepinephrine, sympathomim, epinephrine and isoproterenol) and blocking agents (phenoxybenzamine and propranolol) were studied in control (N=55) and exercising (N=52) albino rats under anaesthesia. The test rats exercised by regular swimming for 10-14 weeks. Alpha stimulation and beta blocking produced smaller responses while alpha blocking and beta stimulation were followed by greater changes after training as compared with the control animals. The assumption of a modified adrenergic receptor sensitivity could not be substantiated by the results; the observations indicate rather a complex change in the autonomous regulation following regular physical exercise.


Myocardial scintigraphy with 99mTc-pyrophosphate in 150 coronary care unit patients. 150 patients, admitted to the coronary care unit with suspicion of acute myocardial infarction, received pyrophosphate labelled with 99mTc, 6-120 h after onset of symptoms, mean 24 h, and were examined in the anterior posterior position and in the left anterior oblique position with a mobile gamma camera. Scintigrams were obtained initially at the injection, and then every 15th min during 1h. The scintigrams were evaluated with regard to presence and localization of radionuclide uptake in myocardial area. In 98 patients with a clinical diagnosis of acute myocardial infarction, uptake was found in 95, with good correlation between ECG and scintigraphic localization. 2. patients with myocardial infarction, verified at autopsy, did not show any uptake and 1 patient, surviving the myocardial infarction, also showed negative result. 19 of 26 patients with unstable angina pectoris also exhibited an uptake in the myocardium. 25 of 26 patients with other diagnoses showed no uptake, while in 1 patient an uptake was recorded. It is concluded that with 99mTc-pyrophosphate scintigraphy it is possible to separate ischemic heart disease from other diseases in patients with chest pain.


Hyperlipoproteinemia in angiographically documented coronary artery disease. Plasma lipids and lipoprotein levels were measured in 100 patients who underwent coronary angiography for chest pains. The coronary arteries were normal in 33 patients (22 males and 11 females), moderately pathological (30--60% narrowing) in 4, and markedly pathological (greater than 60% narrowing) in 63 (53 males and 10 females). Plasma cholesterol and triglyceride levels and very low and low density lipoprotein cholesterol levels were higher by 20--40% in both males and females with pathological coronary arteries as compared to normals. The high density plasma lipoprotein cholesterol level was lower in females with pathological arteries than in normals, but did not differ between the male groups. 67% of the males with pathological arteries had hyperlipoproteinemia as compared to 26% of the normals, and hyperlipoproteinemia was as frequent among males emigrating to Isreal from Europe or America as among those born in Asia or Africa. Hyperlipoproteinemia types IIA, IIB and IV were encountered among the patients at similar rates.


Antigenic specificity of the cytolytic T lymphocyte (CTL) response to murine sarcoma virus-induced tumors. I. Preferential reactivity of in vitro generated secondary CTL with syngeneic tumor cells. Incubation of spleen cells from mice having rejected a Moloney sarcoma virus (MSV)-induced tumor with syngeneic irradiated lymphoma or sarcoma cells bearing MSV-associated antigens in secondary mixed leukocyte-tumor cell cultures (MLTC) resulted in the generation of highly active cytolytic T lymphocytes (CTL) specifically directed against syngeneic target cells bearing MSV-associated antigens. When MSV-immune spleen cells from C57BL/6 (H-2b) and BALB/c(H-2d) mice were compared with respect to their ability to generate CTL in syngeneic secondary MLTC, it was found that both lymphoid cell populations were equally able to mount an anamnestic CTL response to MSV-associated antigens as assessed by a short-term 21Cr release assay. However, quantitative analysis of the activity of both CTL populations on either H-2b or H-2d tumor cells indicated that target cells sharing the same major histocompatibility complex (MHC) as the effector cells were lysed 10- to 100-fold more efficiently than allogeneic target cells. As suggested by the results of inhibition experiments using mixtures of 51Cr-labeled and unlabeled target cells, preferential lysis of syngeneic versus allogeneic tumor cells might be related to the establishment of effective adhesions between the former and CTL. Direct evidence for the role of MHC in determining the antigenic specificity of CTL directed against MSV-associated antigens was provided by results obtained using MSV-immune spleen cells from congenic resistant mice. Furthermore, studies of the response of F1 (H-2b/d) hybrid mice showed that stimulation of immune spleen cells with tumor cells from one parental strain or the other in secondary MLTC resulted in the generation of CTL capable of lysing tumor target cells of the same perental strain as the stimulating cells, but not of the other. The results thus suggested the presence of two sets of CTL precursor cells in F1 MSV-immune spleens, each set responding exclusively to tumor antigens associated with only one of the two parental phenotypes.


Differences in effect of isoproterenol stimulation on levels of cyclic AMP in human B and T lymphocytes. The level of cyclic AMP (cAMP) in human lymphocytes in the presence or absence of isoproterenol stimulation was studied in various lymphocyte subpopulations containing different proportions of B and T lymphocytes. Lymphocytes from thymus and peripheral blood (which contain a majority of T cells) were more sensitive to isoproterenol action than lymphocytes from tonsils or adenoids (where B cells are predominant). Using purified B or T lymphocytes from peripheral blood, tonsils or adenoids, we observed an absence of B lymphocyte response to isoproterenol, whereas T lymphocytes, which had a lower basal c-AMP level than B cells, exhibited in all experiments a significant increase in cAMP content after isoproterenol stimulation. The intensity of the response varied in the different T lymphocyte subpopulations.


Identification of murine leukemia viral antigens detected on mouse cells by H-2 alloantisera. H-2 alloantisera have been previously reported to contain antibodies against murine leukemia viral antigens, but the nature of the viral antigens on mouse cells which interact with these antibodies has not been established. We have found that H-2 alloantisera recognize components of molecular weight 70 000-80 000 mouse lymphocytes and leukemia cells. These components were also detected by a goat antiserum against the murine leukemia virus (MuLV) glycoprotein (gp 70) and are therefore closely related to or identical with that viral protein. Although most H-2 alloantisera detected gp 70-like molecules  on lymphocytes and leukemia cells from a great variety of mouse strains, only one H-2 alloantiserum was found to interact with a gp 70 component on cells from C57BL/10 and C57BL/6 mice. Animals such as C57BL/10 mice that lacked the component reacting with most H-2 alloantisera showed increased serum levels of anti-MuLV antibodies after injection of B10.A spleen cells having a gp 70 component detectable by other H-2 alloantisera. In contrast, strains with cells reactive to antiviral antibodies in the H-2 alloantisera had low responses to MuLV antigens after a similar immunization procedure. Serum levels of anti-MuLV antibodies in both groups of mice, however, were increased after injection of Freund's adjuvant. These observations suggest that anti-MuLV antibodies in mouse alloantisera may arise from a response to viral antigens on the immunizing cells and general stimulation of the immune system.


Interference of inhibitors of dopamine-beta-hydroxylase with uptake of monoamines by chromaffin granular membranes. Disulfiram and bis-(4-methyl-1-homopiperazinylthiocarbonyl)-disulphide (FLA 63) markedly inhibited the Mg2+/ATP-dependent uptake of various monoamines, e.g. dopamine (DA), by isolated membranes of bovine adrenal chromaffin granules. Both compounds affected DA-beta-hydroxylase (DBH) activity more markedly than the uptake of DA. Other inhibitors of DBH, e.g. fusaric acid and diethyldithiocarbaminate (DDC), did not interfere with DA uptake. Disulfiram and FLA 63, in contrast to fusaric acid and DDC, also caused a partial inhibition of Mg2+-dependent ATPase. It is concluded that the inhibition of monoamine uptake by disulfiram and FLA 63 is not related to their effect on DBH.


The effect of adrenergic agonists and cyclic AMP on the Na+/K+ ATPase activity of brown adipose tissue. The Na+/K+ ATPase activity of membrane fractions prepared from brown adipose tissue of cold-acclimated rats could be stimulated by addition of any one of the following: norepinephrine, isoproterenol, cyclic AMP or phenylephrine. These results are consistent with the proposal that in the intact brown adipocyte, the norepinephrine-induced stimulation of the Na+/K+ membrane pump is associated with alpha- as well as beta-adrenergic pathways and is, in part, mediated via cyclic AMP.


The role of prostaglandins in cholinergic neurotransmission in the guinea pig. Prostaglandins have contrasting effects on neurotransmission at different cholinergic nerve endings. This is a report on the role of prostaglandins in a number of cholinergic preparations from the guinea pig. In the isolated ileum PGE1 (2 X 10(-10) to 5 X 10(-8) M) potentiated the response to electrical stimulation of the cholinergic nerves. PGE1 (10(-7 M) caused an increase in tone followed by a period of transient inhibition of twitch height. Responses to simulation of the ileum with drugs were not potentiated by PGE1. Responses of atropinized or plexus-free muscle to electrical stimulation were also not potentiated by PGE1. Acetylsalicylic acid (2.5 X 10(-4) M) diminished the twitch response and the output of acetylcholine from the ileum. Both effects were reversed by PGE1. Qualitatively similar observations were made on the trachea. It is concluded that prostaglandins facilitate acetylcholine release in the ileum and trachea. PGE1 diminished the effect of vagal stimulation on the heart rate. The response to stimulation of the phrenic nerve was not affected.


Differences between the effects of dopamine and apomorphine on rat aortic strips. Dopamine and apomorphine have been compared with regard to contraction and relaxation of aortic strips prepared from rats of different ages. Both dopamine and apomorphine contracted aortic strips from older (9-12 weeks) rats to greater maximal tension than preparations from younger (3--5 weeks) animals. Contraction in response to both agonists could be blocked by the alpha-blocker, Dibenamine. In contrast to dopamine contraction by apomorphine was associated with the development of tachyphylaxis. Both apomorphine and dopamine relaxed the aorta after alpha blockade, however, the relaxation produced by these drugs differed in two major aspects. First, dopamine-induced relaxation was greatest in aortic strips from young rats compared to older rats whereas relaxation with apomorphine was not age dependent. Second, dopamine-induced relaxation was abolished by propranolol but not haloperidol whereas apomorphine relaxation was only blocked by haloperidol. These data establish that the aortic relaxation caused by dopamine is most likely a beta-adrenergically mediated response whereas that produced by apomorphine is not. Moreover, only dopamine was able to increase the concentration of rat aortic cyclic AMP. Should these data be applicable to other vascular beds or species, it is possible that the vascular effects of dopamine will be influenced by age and drugs which impinge on cyclic nucleotide disposition.


Further study of the adrenoceptors of the saphenous vein of the dog: influence of factors which interfere with the concentrations of agonists at the receptor level. In the present study the affinities of some sympathomimetic amines for alpha- and beta-adrenoceptors of the dog saphenous vein tissue were determined after all known factors interfering with the concentration of these agonists at the receptor level had been assessed and excluded. It was observed that in control experiments the relative potencies of sympathomimetic agonists for inducing contractions were: adrenaline (1.6) greater than noradrenaline (1.0) greater than phenylephrine (0.38) greater than isoprenaline (0.009). The elimination of neuronal uptake by cocaine, 4 X 10(-6) M, enhanced predominantly the effects of noradrenaline (by a factor of 7.5), whereas block of catechol-O-methyl transferase (COMT) by U-0521, 10(-4) M, only enhanced those of adrenaline (by a factor of 2.6) and block of beta-adrenoceptors by propranolol, 5 X 10(-7) M, enhanced those of isoprenaline (by a factor of 3) and adrenaline (by a factor of 1.8). Block of COMT enhanced the effects of adrenaline approximately as much as did the blockade of neuronal uptake; this seems to indicate that the affinity of adrenaline for extraneuronal and neuronal uptake processes is approximately the same. Regarding the relaxation-inducing capacity of sympathomimetic agents it was observed that isoprenaline, adrenaline and noradrenaline are full agonists, whereas phenylephrine was not able to produce relaxation amounting to more than 5% of the maximum. Denervation did not modify the relaxant effects of isoprenaline. After elimination of all known factors interfering with the concentration of the sympathomimetic agonists in the biophase, the ratios between the ED50's of each amine for alpha- and beta-adrenoceptors were: adrenaline = 34, noradrenaline = 109 and isoprenaline = 0.0041.


Potentiation of excitatory synaptic transmission in the normal and in the reinnervated dentate gyrus of the rat. Following destruction of the ipsilateral temporo-ammonic tract, which originates in the entorhinal cortex, and terminates on the granule cells of the dentate gyrus, fibers from the surviving contralateral entorhinal area proliferate forming extensive new connections with the denervated dentate granule cells. Utlizing extracellular recording techniques, we have compared the characteristics of synaptic transmission in the lesion induced afferents with the characteristics of the normal ipsilateral afferents by analyzing the responses of dentate granule cells to paired pulse activation of temporo-dentate circuitry. In the dentate gyrus of the normal rat, and extracellularly recorded EPSP evoked by stimulation of the ipsilateral entorhinal cortex is enhanced by as much as 100% by a "conditioning" pulse to the same afferent system. This is called paired pulse potentiation. In the reinnervated dentate gyrus, the extracellular EPSP evoked by a test stimulus delivered to the contralateral entorhinal cortex is also potentiated by a conditioning pulse. Thepaired pulse potentiation in the reinnervated dentate gyrus has a time course which is comparable to that of the normal ipsilateral afferent system, but the magnitude of the potentiation is somewhat less, averaging approximately 140% of control...


Ultrastructure of cultured fibroblasts in mucolipidosis type IV. Five patients have been classified as suffering from mucolipidosis type IV on the basis of the findings in various tissues and in cultured fibroblasts. The light and electron microscopic features of cultured fibroblasts in this disease are described in detail. The differentiation of this condition from other lysosomal storage diseases and other types of mucolipidoses is discussed. The importance of identifying this disease for purposes of intrauterine diagnosis and eventual understanding of the underlying enzyme defect is stressed.


Cyclic AMP binding to intracellular receptor proteins in rat myometrium. Effect of epinephrine and prostaglandin E1. In estrogen-pretreated rat myometrium, the relaxing effect exerted by theophylline or epinephrine has been correlated with their ability to raise cyclic AMP levels (Vesin and Harbon, 1974). The present study demonstrates that such a correlation can be quantitatively extended to the degree of saturation of intracellular cyclic AMP receptors. The rise in cyclic AMP induced by theophylline and/or epinephrine in intact myometrial strips was accompanied by a decrease in the ability of the corresponding extracts to bind exogenous 3H-labeled cyclic AMP. Total intracellular cyclic AMP binding sites were not modified and averaged a value of 0.22 muM. Accurate estimation of intracellular receptor-cyclic AMP complex has been correlated with the corresponding level of cyclic AMP in the tissue, the apparent intracellular Kd for cyclic AMP has been evaluated at 450 nm. Stimulation of myometrial strips with prostaglandin E1 (PGE1) which has been shown previously to induce contractions, although elevating cyclic AMP levels, was accompanied by a parallel increase in the saturation of the endogenous receptor, in an identical manner to that found with epinephrine or theophylline. The postulated hypothesis for a compartmentalization of cyclic AMP, or an interference of PGE1 with the intracellular cyclic AMP binding equilibrium has not been verified. The cyclic AMP system cannot be considered as the exclusive mechanism regulating uterine relaxation.


Ontogenesis, distribution and relative sensitivity of hCG receptors to hCG and LH in goat ovaries. The binding of [125I]hCG to immature and mature follicles and corpora lutea of goat ovaries has been studied. The hormone is bound maximally by corpora lutea although mature follicles also exhibit some binding. Immature follicles are practically devoid of receptors for this hormone. In the corpus luteum, the receptors for the hormone are present in thecal and luteal cells. Autoradiographic studies show the location of the bound radioactivity grains primarily along the plasma membranes of these cells, although some radioactivity grains were also seen in the cytoplasm of luteal but not thecal cells. On a mole to mole basis, hCG was found to displace [125I]hCG from binding to receptors on corpus luteum better than hLH and oLH.


The use of a multicomponent standard for clostridial vaccines. A multicomponent clostridial vaccine in being developed with the objective of acting as a standard preparation for the control of vaccines containing one to eight components. It is intended initially for use in monitoring those components which can be assessed by antitoxin response assays in rabbits. The components involved, therefore, are the principle toxoids of Clostridium (perfringens) welchii types B, C and D, Cl. oedematiens type B, Cl. septicum and Cl. tetani. The proposed standard also contains the toxoid of Cl. oedematiens type D and an anaculture of Cl. chauvoei. It is hoped later to develop the use of the two latter components as standards in other test systems. The aim of establishing the standard was to provide a means of controlling inter-laboratory variation, especially that due to animals.


Effects of trypsin and phospholipases A2 and C on enzyme organization in testis microsomes. Pregnenolone and progesterone concentrated in the microsomal fraction of cryptorchid mouse testis compared with mitochondria and cytosol. While the concentrating mechanisms had high capacity and low association constants the effect did not seem to be due to nonspecific solubility in the lipid components since 17-hydroxyprogesterone, dehydroepiandrosterone, androstenedione and testosterone did not show differential concentration. Also digestion with phospholipases A2 and C to the point where most of the phospholipids were specifically split, only lowered the differential binding of pregnenolone and progesterone by less than half. Trypsin had a greater effect, short digestion at 0 degrees C lowering the specific binding to 35-40% and decreasing the steroid dehydrogenases to a similar extent. The members of the mixed function oxidase system in the testis microsomes were particularly sensitive to trypsin, cytochrome P-450 and, as a consequence, 17alpha-hydroxylase and 17, 20-lyase activity being eliminated under tha same conditions while liver microsomal cytochrome P-450 was hardly affected. Bonds split by trypsin seem to play a more important role in the hydroxylase activity of testis microsomes than in the hepatic system.


An international survey of clostridial sera and vaccines. The papers present the results of a survey of the usage, assay and specification of veterinary clostridial sera and vaccines in 23 countries. Thirteen of the countries use up to 8 different antisera. All the countries use vaccines, which are prepared from 13 species and types of clostridia. Vaccines containing up to 8 such components are commonly employed. Criteria for the design and interpretation of assays are discussed and evidence for efficacy summarized.


Production and standardization of polyvalent Clostridium perfringens vaccine in Iran. In this presentation an account of the large-scale production and the standardization of polyvalent Clostridium perfringens vaccine in Iran is described. Over 20 million doses of this vaccine are produced and utilized in Iran every year. Certain modifications have been made to the culture media used in our laboratory for the production of this vaccine in order to bring down its cost. The prepared vaccine is highly immunogenic as determined by the laboratory examination on the quality of the vaccine according to British Veterinary Codex and the field reports.


The standardization of Cl. perfringens antigens and antisera. The preparation of laboratory standard antitoxins against Cl. perfringens beta and epsilon toxins is described. These antitoxins are suitable for the quantitative determination of the corresponding antigens by means of the flocculation test. The flocculation test was, however, shown to be more suitable for determining the antigenic value of fresh toxoid rather than toxoid stored without neutralization of excess formalin. A maximal immunity response to alum-precipitated epsilon toxoid was obtained in sheep with two injections containing 90 Lf per dose. The interval between these injections may vary from 2 to 6 weeks. The serum-antibody titres after the primary and secondary injections or after a booster dose given before 12 months after the primary injection did not remain above the protective level in most of the sheep injected for longer than about 5 months. When a sound basic immunity is established the degree of protection following on a booster dose given 12 months later is complete for at least 12 months. An alum-precipitated vaccine containing 25 Lf epsilon toxoid per dose is adequate. The decline in the serum-antibody titre during the first year of vaccination could be eliminated by the use of the antigen in water-in-oil emulsion. Lambs from immune dams were protected for at least up to 13 weeks of age. A satisfactory level of circulating antitoxin against Cl. perfringens beta toxin could be produced in ewes by vaccinating them with APT containing 6.25 Lf beta toxoid per dose. The primary and secondary injections could be separated by 2, 3, 4 or 5 weeks without changing the end result. A booster dose given 2 months before parturition was satisfactory.


[Standardization of the control technics of Clostridium perfringens vaccine. Use of a reference vaccine]. Control of Cl perfringens toxins. It is possible to employ a standardized technique for the control of the toxins of Cl. perfringens. Titres of LD50 per ml for the mouse and L+/50 per ml may be expressed with a confidence limit of 1.5. The assay of toxins by L+/50 (relating to their antigenic value) permits the establishment of standards by which it is possible to obtain a satisfactory vaccine with a high degree of certainty. 10 L+/50 per dose for Cl. perfringens A 20 L+/50 per dose for Cl. perfringens B 180 L+/50 per dose for Cl. perfringens C 50 L+/50 per dose for Cl. perfringens D Control of vaccine values. Technique adopted : Determination of antibodies in the rabbit following vaccination. The confidence limit of the antitoxin titre is similarly equal to 1.5 when P = 0.05. It would not appear to be necessary to establish minimum levels of antibodies to confer protection; it seems nevertheless that the results are influenced by considerable variations attributable to the animals used for the most part and that a reference vaccine is required. Freeze-dried vaccine. There is a natural antigenic activity in the rabbit which retains a considerable part of its antigenic properties after maintenance for one week at +37 degrees C. In our view this would be suitable for a reference vaccine.


[Comparison between the official indirect method and the direct method of control of vaccines against sheep enterotoxemia]. The official methods of control of vaccines against sheep enterotoxemia are indirect in two ways : on the one hand they are carried out on animals for which the vaccine is not intended, and on the other hand they test only an immunological serum reaction. Another method consisting of direct testing by intravenous injection of toxin has been carried out on mice previously vaccinated with different doses. It has been shown that it is possible to obtain a certain level of protection giving a good dose-effect relation. However, immunity provided by this direct method is weak although the vaccine utilized is considered very efficacious with regard to the official norms of the direct test. These results, which are both encouraging and disappointing, will be the subject of a more intensive study of the different parameters in question.


Experimental studies on serum treatment and vaccination against Cl. perfringens type C infection in piglets. Subcutaneous administration of serum to piglets just after birth resulted in serum titres of 9-18 I.U. beta-antitoxin per ml in the first three days of life. At the ages of 7, 14, and 28 days the titres had dropped to about 5-9, 2 and 1 I.U. per ml, respectively. Oral administration of the same dose of serum resulted in serum titres of about half of those found after s.c. administration. In infected herds a significant protective effect after both s.c. and oral administration of serum was found to be dependent on the time of treatment but independent of the route of administration. After vaccination a correlation was noted between the levels of beta-antitoxin in colostral whey and specific mortality in the litters. An initial beta-antitoxin concentration of about 10 I.U. per ml whey seems to be sufficient to secure effective prevention. By vaccination once during gestation the beta-antitoxin levels in colostral whey were all less than 10 I.U. per ml. Two vaccinations during gestation resulted in whey titres greater than 10 I.U. per ml in 12 of 20 dams. By revaccinating once only during the following gestation effective beta-antitoxin levels in colostral whey were secured regardless of whether the vaccination had been performed once or twice during the previous gestation : the mean was 87.4 I.U. beta-antitoxin per ml; three of 20 dams had titres less than 10 I.U. per ml whey. From mortality studies including 63 liters in three infected herds specific mortalities of 17.3% and 4.6% were found after one and two vaccinations respectively, as compared with 36.6% in the control group. After revaccination during the ensuing gestation the figures were 1.4%, 0.0% and 32.2% named in the same order. 2 ml serum given as soon as possible after birth or 5 ml vaccine injected twice during gestation followed by one revaccination during subsequent gestations effectively protect piglets against infection with Cl. perfringens type C.


Solid-phase radioimmunoassays for quantitative antibody determination of bacterial exotoxins. Measurement of Clostridium perfringens type D epsilon antitoxin. Two reversed solid-phase radioimmunoassays have been developed for quantitative determination of antibodies against Clostridium perfringens type D epsilon toxin. 125I labelled prototoxin was used in the bromoacetyl cellulose-bound antibody method and in the antibody coated tube method. The procedures are based on the competition for 125I labelled prototoxin between the insoluble antibodies and the antibodies present in the unknown sample. The radioactivity bound to solid-phase is in inverse ratio to quantity of measured antibodies. The antibody values which can be detected are in the range of 0.004 IU/ml of investigated serum. The methods allow the rapid and inexpensive screening of large groups of vaccinated sheep, and are very suitable for measuring small amounts of Cl. perfringens D epsilon antibodies with a small experimental error.


An international serotyping system for Clostridium perfringens (welchii) type A in the near future. The importance of Clostridium perfringens (welchii) type A as an agent of clinical infection in man is well known and, more recently, its role as a cause of human food poisoning has become well recognized. There has been a growing awareness of the need for a system of finer subdivision within the type A group for better understanding of the epidemiology of Cl. perfringens in infections and food poisoning. A set of antisera used for typing and developed in the Food Hygiene Laboratory over a period of some years has proved to be of considerable value in this respect; it is now being integrated with similar sets of antisera prepared independently in the USA and Japan to form a comprehensive and internationally workable serotyping scheme. Negotiations regarding commercial production of the main type antisera are now in progress. It is hoped that serotyping facilities will be available in the near future to workers and institutions concerned with Cl. perfringens type A.


Postheparin plasma lipoprotein lipase and hepatic lipase in diabetes mellitus. Relationship to plasma triglyceride metabolism. The activity of two triglyceride lipases was determined by an immunochemical method in the postheparin plasma of 60 diabetic patients and of 47 age- and sex-matched nondiabetic control subjects. The results were related to the type of diabetes, to plasma triglyceride and insulin concentrations, to removal of exogenous fat from the blood, and to turnover of VLDL-triglycerides . The mean postheparin plasma lipoprotein lipase (LPL) activity was decreased by 44 per cent (p less than 0.001) in patients with untreated ketotic diabetes and by 20 per cent (p less than 0.01) in patients with untreated mild to moderate nonketotic early-onset diabetes. Insulin treatment of ketotic diabetes resulted in a rapid increase in the activity of LPL and decrease in serum triglycerdie level, whereas sulfonylurea treatment of non-insulin-requiring diabetics did not significantly influence the enzyme activity. In insulin-treated chronic diabetics the average postheparin plasma LPL activity was not different from that of nondiabetic controls, but some of these patients had high LPL values. In normolipidemic maturity-onset-type diabetics the LPL activity was within normal range, but in those having hypertriglyceridemia the average LPL value was decreased by an average of 26 per cent (p less than 0.01). The LPL activity showed a significant negative correlation with the logarithm of serum triglyceride concentration (r = -0.62) and a positive correlation with fractional removal of Intralipid (r = +0.64) and fractional turnover of V triglyceride (r = +0.40). The activity of LPL was correlated to basal plasma insulin concen tration in the insulin-deficient diabetes r = +0.34) but not in patients with maturity-onset-type diabetes. The hepatic lipase (HL) activity of postheparin plasma was similar in diabetes and controls, with the exception of hypertriglyceridemic maturity-onset diabetics, who had higher mean HL activity than the corresponding control group (p greater than 0.01). The activity of HL was not related to triglyceride removal but showed a significant correlation to VLDL-triglyceride production rate. On the basis of these results it seems that a deficiency of LPL accounts for a great deal of the elevation of serum triglyceride in insulin-deficient human diabetes but has a smaller role in the pathogenesis of the hypertriglyceridemia that is associated with maturity-onset diabetes. The latter abnormality is caused mainly by an increased secretion of triglycerides into the blood even though a decreased LPL may contribute to development of hyperlipemia in cases with gross elevation of serum triglycerides.


Role of insulin resistance in adipose tissue and liver in the pathogenesis of endogenous hypertriglyceridaemia in man. Studies were performed to evaluate the relative importance of enhanced adipose tissue lipolysis and increased insulin levels in modulating hepatic VLDL production in patients with endogenous hypertriglyceridaemia. Eight control subjects and nine patients with hypertriglyceridaemia were investigated. The latter group comprised four patients with idiopathic hypertriglyceridaemia, three maturity onset diabetics, and two siblings with diabetic lipodystrophy. Each individual's plasma VLDL was selectively labelled with I131 in the apoprotein moiety and then reinjected to assess the turnover of these molecules. This was correlated with the insulin response to an oral glucose load and with the plasma FFA flux measured by a continuous infusion of 14C palmitate. In the patients with idiopathic hypertriglyceridaemia and in the adult onset diabetics, plasma VLDL-appoprotein turnover was increased suggesting enhanced hepatic production of these molecules. Although the insulin levels in these patients were higher than normal, no significant correlation was demonstrable between the plasma insulin and the turnover of VLDL-B-apoprotein. Furthermore, in the two patients with lipo-dystrophy the turnover of plasma VLDL was within the normal range, whereas the plasma insulin responses were the highest among all the patients. These results suggest that hyperinsulinaemia alone is not sufficient to account for the increased VLDL production seen in some of our patients. The plasma FFA flux was raised in the patients with idiopathic hypertriglyceridaemia and in the maturity onset diabetics, and was within the normal range in the two patients with lipodystrophy. Indded, in all the subjects studied a significant correlation was observed between the turnover of plasma VLDL-B-apoprotein and the plasma FFA flux. The results thus indicate that the rate of FFA release to plasma constitutes the predominant factor in determining hepatic output of VLDL and that in the majority of patients with endogenous hypertriglyceridaemia the increased FFA flux resulting from insulin resistance in adipose tissue could effectively increase VLDL production. This process appears to be independent of the prevailing insulin levels, and could occur in the presence of insulin resistance in the liver. The latter, however, could be responsible for the impaired glucose tolerance observed in some patients.


Decreased cyclic AMP and insulin responses to glucose in pancreatic islets of diabetic Chinese hamsters. The dose as well as the time kinetics of insulin and adenosine-3',5'-monophosphate (cyclic AMP) responses to glucose were compared in pancreatic islets isolated from normal and diabetic Chinese hamsters. The insulin content in diabetic islets was about one-half that in normal islets. Insulin release in diabetic islets incubated for 10 min with glucose 60-1000 mg/100 ml was from one-third to one-half that in normal islets. Glucose 1000 mg/100 ml stimulated three-fold increases in insulin release without increasing the accumulation of [3H] cyclic AMP in either normal or diabetic islets prelabelled with [3H] adenine. However, in the presence of 1.0 mM of the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IBMX), glucose 150 mg/100 ml elicited significant increases of insulin release (+ 134%) and [3H] cyclic AMP accumulation in islets (+ 44%) and incubation medium (+ 48%) of islets of normal but not diabetic hamsters. Also, in perifusion experiments with 0.1 mM IBMX, glucose 500 mg/100 ml produced threefold greater increases in insulin release and two-fold greater increases in efflux of cyclic AMP in normal than diabetic islets. By contrast with the lesser effects of glucose in diabetic islets, 1.0 mM IBMX increased islet and medium cyclic AMP, as well as insulin release, similarly in normal and diabetic islets. It is suggested that the impairment of glucose-induced insulin release in islets of the diabetic Chinese hamster may be due to a defective interaction of glucose with the adenylate cyclase-cyclic AMP system in the pancreatic B cell.


Abnormalities in the adrenergic control and the rate of lipolysis in isolated human subcutaneous adipose tissue in diabetes mellitus. Subcutaneous adipose tissue was obtained from 9 patients with untreated diabetes mellitus and from 13 obese nondiabetics. After incubation with isoprenaline or noradrenaline, glycerol release and tissue cyclic AMP (cAMP) were determined. Basal glycerol release was twice as rapid from the diabetic adipose tissue. With isoprenaline, the cAMP concentration and the glycerol production was significantly higher in the diabetic adipose tissue. Noradrenaline did not increase glycerol production or cAMP concentration in the diabetic adipose tissue. Subcutaneous adipose tissue was also removed from the diabetics after antidiabetic treatment. Basal lipolysis was significantly reduced and noradrenaline significantly increased both glycerol release and cAMP production. With isoprenaline, cAMP production and glycerol release were significantly less after antidiabetic treatment than in the untreated state. The data provide evidence for increased alpha- as well as beta-adrenergic receptor sensitivity in human subcutaneous adipose tissue of untreated diabetic patients.


Some properties and electrophoretic patterns of rat liver esterases in relation to hepatocarcinogenesis by 3'-methyl-4-(dimethylamino) azobenzene. Five distinctly different types of naphthyl acetate esterase in rat liver were examined for study of liver enzymes during hepatocarcinogenesis. Three types of esterase in normal adult liver were separated by column chromatography. Main esterase in adult hepatocytes, which was demonstrated near the origin in cellulose acetate electrophoresis, was very sensitive to diisopropyl phosphorofluoridate (DFP). The other two esterases showed different electrophoretic mobility, while their Km values did not differ and both were considerably resistant to DFP. An anodic minor component in normal adult liver, which had a characteristic esterase pattern of infant liver, increased in the liver of rats fed 3'-methyl-4-(dimethylamino) azobenzene. This esterase was obtained by electrophoresis on Cellogel block. It was partially inhibited by DFP and p-chloromercuribenzoate, activated by cysteine, and showed a different Km value from the above esterases. Another minor component situated at the most cathodic side, which had characteristic esterase patterns of fetal liver and hepatoma, was very sensitive to DFP and eserine, and showed a characteristic of nonspecific cholinesterase as proved by staining.


Type-C RNA viruses and leukemogenesis: relation of type-C virus infectivity and leukemogenesis induced by nitrosourea compounds in mice. Correlation between infectivity of type-C RNA virus) murine leukemia virus, MLV) and development of leukemia was tested  in female ICR/JCL mice treated with either 1-ethyl-1-nitrosourea (ENU) or 1-butyl-1-nitrosourea (BNU). Continuous administration of either chemicals resulted in the occurrence of thymic lymphoma in every mouse with a short latent period. The time of appearance and distribution pattern of MLV infectivity in various tissues were examined by the XC plaque assay technique at fixed intervals during the leukemogenic treatment. In ENU- or BNU-treated mice, only a few samples of the thymus showed MLV infectivity with rather low titers during incubation period and the presence of MLV was not consistent even in leukemic cases though the thymus was almost invariably the target of leukemogenesis. On the other hand, many samples of the uterus, spleen, and mesenteric node from non-leukemic and leukemic mice harbored a good quantity of MLV. In tissues such as the liver, kidney, bone marrow, and muscle, positive cases occurred only sporadically. Observations on the MLV infectivity in untreated controls were almost comparable with those in leukemogen-treated mice. These results indicate that the infectivity of MLV, detected by the XC plaque assay technique, is not necessarily related to the induction of leukemia in mice by exogenous agents.


An experimental model for lymphatic metastasis in rats. An experimental model was described for inducing spontaneous lymph node metastasis, in which tumor cells of the rat ascites hepatoma AH-109A were transplanted into the subcutaneous tissue of the penis, which is rich of lymph plexus. Growth of tumor at the site of transplantation was measured serially and metastasis was recognized by palpation in the inguinal lymph nodes initially, and then in the axillary and lumbar nodes. When animals were sacrificed 12 days after transplantation, these lymph nodes were weighed and high percentage of metastasis was confirmed by histological examination. As an experimental model of lymph node metastasis, the usefulness of this method was discussed.


Viral DNA in inflammatory bowel disease. CMV-bearing cells as a target for immune-mediated enterocytolysis. Althouth serological studies have revealed a high incidence and mean titer of antibody to cytomegalovirus (CMV) in some patients with inflammatory bowel disease, electron microscopy has not confirmed the presence of mature virus in diseased tissue. Sensitive biochemical techniques for detection of viral nucleic acid were applied therefore to test the proposed disease-virus association. Membrane cRNA-DNA hybridization showed that CMV DNA was not detectable in 3 cases of Crohn's disease, nor was it present in 3 or 4 cases of ulcerative colitis examined. Five tissue specimens taken 5 cm away from a colonic tumor were also negative. An in situ hybridization study of diseased tissue, including specimens from 6 inflammatory bowel disease patients in addition to those above, showed a few grains distributed over cells superficial to the muscularis mucosa, but was not regarded as significantly different from controls. The association of CMV with Crohn's disease (0 positives in 4 patients by either test) is not supported by this study; the association with ulcerative colitis (1 positive in 9 patients by either test) requires further study before a definitive conclusion can be made.


Rat gastric mucosal adenylyl cyclase.

Rat gastric mucosal adenylyl cyclase. Prostaglandin E1, epinephrine, secretin, and glucagon are known inhibitors of gastric acid secretion, and each agent stimulated mucosal membrane (600 X g pellet) adenylyl cyclase activity from the corpus of the rat stomach. This adenylyl cyclase activity was also stimulated by 5'-guanylyl-imidodiphosphate and sodium fluoride but not by guanosine-5'-triphosphate. By contrast, the gastric acid secretagogues, pentagastrin, histamine, and carbachol, had no effect on basal or prostaglandin E1-stimulated mucosal adenylyl cyclase activity. Most of the sodium fluoride- and hormone-stimulated adenylyl cyclase of the corpus mucosa was contained in the 600 X g membrane fraction. The enzyme exhibited Michaelis-Menten kinetics with respect to the concentration of ATP, with an apparent Km of 0.25 mM. Histamine did not stimulate rat mucosal adenylyl cyclase activity under a variety of conditions, but did stimulate the same enzyme in guinea pig gastric fundic mucosa, an enzyme also activated by prostaglandin E1. These studies do not support the hypothesis that cyclic AMP mediates the actions of gastric acid secretagogues on the parietal cell in the rat.


Activation of rat gastric mucosal adenylyl cyclase by secretory inhibitors. Prostaglandin (PG) E1, E2, A1, and A2 stimulated rat gastric corpus mucosal membrane adenylyl cyclase activity. PGE1 (Kalpha congruent to 8 muM) affected the maximum velocity but not the affinity of the enzyme for ATP and maximum PGE1 activation was not affected by histamine H1 or H2 receptor antagonists. 5'-Guanylyl-diphosphoimide (Gpp(NH)p), but not GTP, stimulated both the basal and PGE1-stimulated adenylyl cyclase activities, although the percentage stimulation by maximal PGE was the same with or without Gpp(NH)p. NaF stimulation was also additive to that of PGE1. Secretin also stimulated gastric mucosal adenylyl cyclase activity (Kalpha congruent to 30 nM). Maximal secretin activation was not additive to that of PGE1, suggesting a coupling to the same adenylyl cyclase catalytic site. These studies suggest that mucosal membranes may contain beta-adrenergic receptors. The adenylyl cyclase activating agents used in this study, PGE1, secretin, and the catecholamines, are all known inhibitors of gastric acid secretion, suggesting a possible involvement of cyclic AMP in the inhibition of acid secretion in the rat stomach.


Canine Zollinger-Ellison syndrome.

Canine Zollinger-Ellison syndrome. The unusual finding of peptic esophagitis and duodenal ulceration in a dog was associated with a malignant pancreatic islet cell tumor producing gastrin and ACTH. The finding of a gastrinoma in a non-human species introduces the potential for developing an animal model for the study of the protean genetic biochemical, physiologic and metabolic aspects of the Zollinger-Ellison syndrome.