Contractile and biochemical responses of cardiac and skeletal muscle to isoproterenol covalently linked to glass beads. The effects of (-)-isoproterenol covalently linked to glass beads on inotropic state, cyclic AMP concentration, and phosphorylase b to a conversion was studied in cat and guinea pig papillary muscles. Biochemical responses also were measured in mouse diaphragm sections. In cat papillary muscles under normal conditions and in guinea pig muscles partly depolarized with 22 mM K+ an increased inotropic state and phosphorylase activation could be dissociated from the formation of cyclic AMP. This contrasts with close correlation reported between isometric tension and cyclic AMP concentration in guinea pig papillary muscles exposed to varying concentrations of isoproterenol dissolved in the bath medium. The cyclic nucleotide did increase in guinea pig papillary muscles exposed to a freshly prepared batch of isoproterenol beads and in the mouse diaphragms. The type and age of the bead preparation had marked effects on all responses that were measured and on the rate of leakage of biologically active products from the beads. Nevertheless, experimental conditions could be obtained under which the initiation of the contractile and biochemical responses was probably limited to intense stimulation of receptors at or near the points of contact between beads and muscle and not due to gross leakage into the bath. The results indicate that cyclic AMP is probably involved in the initiation but not the propagation of the inotropic response of papillary muscles exposed to isoproterenol glass beads.


Defective calcium transport by cardiac sarcoplasmic reticulum in spontaneously hypertensive rats. Ca2+ uptake and binding and Ca2+-ATPase activity of cardiac sarcoplasmic reticulum (SR) from spontaneously hypertensive rats (SHR) were compared to that obtained from normotensive Wistar-Kyoto (WKY) rats. Ca2+ uptake (172 +/- 3.7 nmol/mg of protein per min in WKY vs. 112 +/- 2.6 in the SHR, P less than 0.001) and binding (154 +/- 3.0 nmol/mg per min in WKY vs. 101 +/- 1.8 in the SHR, P less than 0.01) were decreased in the SHR. Ca2+-ATPase activity, however, was significantly higher in the SHR (118 +/- 3.1 nmol of P per mg of protein per min vs. 86 +/- 1.1 in the WKY, P less than 0.001), suggesting "uncoupling" of the ATPase to calcium transport. Cyclic AMP-dependent phosphorylation of SR was significantly decreased in SHR (0.71 +/- 0.05 vs 0.32 +/- 0.07 nmol of P/mg of protein per 10 min, P less than 0.001) and there was an excellent correlation between cyclic AMP-induced phosphorylation of SR and Ca2+ uptake (r = 0.81). Differences in both cyclic AMP-dependent phosphorylation and Ca2+ uptake between the two groups were evident at 10 weeks and increased progressively to 22 weeks of age. Differences in endogenous cyclic AMP-dependent protein kinase activity may partly explain the decreased Ca2+ transport in SHR.


Cholesterol determination in high-density lipoproteins separated by three different methods. We describe a simplified method for measuring high-density lipoprotein cholesterol in serum after very-low- and low-density lipoproteins have been precipitated from the specimen with sodium phosphotungstate and Mg2+. Values so obtained correlate well with values obtained with the heparin-Mn2+ precipitation technique (r = 0.95, CV less than 5% in 66% of the subjects studied and between 5 and 10% in the remaining ones) or by ultracentrifugal separation (r = 0.82, CV less than 5% in 80% of the subjects studied and between 5 and 10% in the remaining ones). Our precipitation technique is more appropriate for routine clinical laboratory use.


The beta-lipoprotein doublet in type 3 hyperlipoproteinemia. We used a previously reported agarose-gel electrophoresis system to study the lipoprotein patterns in patients with type 3 hyperlipoproteinemia. All samples from 20 subjects tested revealed the presence of two bands with beta-lipoprotein mobility, irrespective of quantitative changes in plasma lipids induced by diet or by the administration of drugs. If these observations are confirmed, the method will permit the diagnosis of type 3 hyperlipoproteinemia without resort to preparative ultracentrifugation and chemical measurement of the very-low-density lipoproteins.


Effect of glucagon infusion on plasma cyclic AMP in patients with cholestatic hepatitis and obstructive jaundice. New test of hepatic cholestasis. Plasma cyclic AMP concentration during glucagon infusion at various time intervals was determined in 8 normal subjects, 9 patients with extrahepatic obstructive jaundice and 10 patients with cholestatic hepatitis (hepatitis A and B). Plasma cyclic AMP concentrations (pmol/ml) during glucagon infusion in patients with both obstructive jaundice and cholestatic hepatitis were found to be greater than those in control subjects. In addition, a significant difference in plasma cyclic AMP concentrations was found between patients with cholestatic hepatitis and obstructive jaundice at the 10th minute of glucagon infusion. These results indicate that plasma cyclic AMP levels at the 10th minute of glucagon infusion represent a reliable diagnostic index of cholestatic jaundice.


Urinary cyclic AMP and post-traumatic acute renal failure. Consecutive daily urinary excretion of cyclic AMP has been investigated in 16 patients with severe trauma or illness, five of whom developed acute renal failure (ARF). Fluctuations in the nucleotide excretion exceeded the range found in 20 healthy volunteers (1.26-14.74, mean 7.13+/-1.18 vs. 2.04-10.10, mean 5.07+/-2.21 micronmol/24 h). This resulted in a 41% increase of cAMP excretion in the group with normal renal function (P less than 0.003) with the highest individual increase of 87%. The excretion usually reached its peak by 24 h after trauma and its lowest value by the third day, (first day vs. third day; 7.82+/-4.23 vs. 3.96+/-2.58 micronmol/24 h, P less than 0.05 for a group of 11 patients), while creatinine clearance remained normal. In four patients with severe ARF, the mean urine volume was above control value but the cAMP excretion was reduced to 3.9 to 14.4% and in one patient with a mild ARF to 60.6%. Creatinine excretion of the group was less reduced than that of cAMP (41.2% vs. 19.6%, resp.). cAMP excretion declined proportionally with diminishing creatinine clearance. In the category of 33-65 ml/min it decreased by 33.4% to 3.39 micronmol+/-1.16 micronmol/24 h. cAMP/creatinine ratio proved to be a less sensitive indicator than cAMP/24 h. Daily output of cAMP and creatinine correlated highly with diuresis in ARF patients, controls (always P less than 0.001) and less in patients with normal renal function (P less than 0.02). Urinary cAMP appears to be a very sensitive and early indicator of the onset of ARF and subsequent recovery. This warrants its further study.


Comparison of plasma hormonal levels between heroin-addicted and normal subjects. The serum concentrations of ACTH, cortisol, aldosterone, thyroxine, cyclic AMP and cholesterol were compared between normal and heroin-addicted subjects. Significantly lower ACTH, cyclic AMP and cholesterol levels were observed to be associated with the heroin addicts, but their plasma thyroxine level was significantly elevated. The possible physiopsychological effect resulting from these changes is discussed.


Electrophoretic mobility in agarose of very low density lipoprotein subfractions in type III hyperlipoproteinaemia. The electrophoretic mobilities in agarose gel of the very low density lipoprotein (VLDL) subfractions of Sf greater than 100, 60-100 and 20-60 from six subjects with type III hyperlipoproteinaemia (HLP) have been compared with those from eight normal volunteers. In type III HLP beta or near-beta (slower VLDL) electrophoretic mobility was not necessarily confined to the VLDL fraction of Sf 20-60 in which it may normally be detected.


Collagenase activity of rat kidney with glomerulonephritis during the heterologous phase. By means of a biological assay the collagenolytic activity of kidneys from rats with Masugi type glomerulonephritis in the heterologous phase and organs from control animals that were given normal heterologous serum was tested. Collagenolysis was found in both examples, but there wre quantitative and qualitative differences in the collagenase activity. In kidneys from animals with Masugi nephritis we found higher activity than in the controls. Normal rat serum inhibits the enzymatic process at a mean percentage of 9.1% whereas EDTA, a selective inhibitor of granulocytic collagenase inhibited the reaction up to 34%. It is concluded that this collagenolytic system is involved in the pathogenesis of the disease by degrading the structural associated collagen of the glomerular basement membrane.


[Activators for sphingohydrolases and the nature of the sphingomyelinase deficiency in Niemann-Pick disease types A, B and C (author's transl)]. Sphingomyelinase activities have been assayed either in the presence of detergents or by an activator method, in control subjects and patients with Niemann-Pick disease types A, B and C. The activity of this enzyme assayed with detergents compared with that for control subjects is markedly reduced in Niemann-Pick disease types A or B but not in type C. With the activator method, the enzymic activity is strongly reduced in patients with type C (30%) of the control data). This paper describes a reasonably satisfactory method for enzymic diagnosis of Niemann-Pick disease type C. Hypotheses for enzymic mutations in these three types of Niemann-Pick disease are put forward.


Low density lipoproteins and immunoglobulins in human pleural effusions. Pleural effusions in patients with bacterial pleurisy contain low density lipoproteins (33 per cent of the blood serum LDL-concentration on average) but almost no very-low density lipoproteins (about 1 percent of the blood serum VLDL-concentration on average). Immunoglobulins are present in large amounts forming a series declining from the IgG (70 per cent of the blood serum concentration, found in effusions) to the IgM (50 per cent of the blood serum concentration). The observations are discussed in relation to morphological aspects of molecular transport through the capillary wall, and the consequences of the LDL- and IgL-accumulation in pleural fluids are briefly discussed.


Thyroxine binding globulin deficiency in a family with type I hyperlipoproteinaemia. A familial type I hyperlipoproteinaemia is described in three members of a family of eleven; on the basis of LPL activity and HDL content of plasma three other members of the family have been diagnosed to be heterozygotes without other disturbances in their lipid spectrum. The distribution of this lipid disorder is in accordance with an autosomal recessive inheritance pattern. In this family a second hereditary condition, thyroxine binding globulin deficiency, was found in addition to the hyperlipoproteinaemia. The inheritance of this condition appears to be as an autosomal dominant. An interrelated inheritance pattern of both conditions could not be proved, but both traits may be located on the same chromosome at some distance from another to allow recombination.


Variable X-linked recessive hypopituitarism with evidence of gonadotropin deficiency in two pre-pubertal males. Two half-brothers with short stature secondary to growth hormone deficiency and a family history implicating X-linked transmission were studied extensively for other endocrine abnormalities. The proband had a normal physical examination, except for small stature and small external genitalia. ACTH and TSH release were normal. LH and FSH responses during an i.v. GnRH test were severely blunted. His half-brother also had a normal physical examination, except for severe short stature and very small external genitalia. Deficiencies of ACTH, and TSH as well as GH were documented. An i.v. GnRH test showed no LH or FSH response. These studies support the existence of an X-linked recessive form of hypopituitarism and portend the clinical usefulness of the i.v. GnRH test in evaluating gonadotropin reserve.


Heterogeneity in maple syrup urine disease: aspects of cofactor requirement and complementation in cultured fibroblasts. Fibroblast strains derived from six patients with maple syrup urine disease have been investigated for their requirements of the cofactors NAD, CoASH, Mg++ and TPP in comparison with 10 normal control strains. The reconstitution of the decarboxylase function of branched chain alpha-keto acid (BCKA) dehydrogenase complex in lysed cells was studied with respect to the substrates alpha-keto-isocaproic acid, alpha-keto-isovaleric acid, and alpha-keto-beta-methylvaleric acid (KIC, KIVA, MEVA). The enzyme activity of all normal control strains for the substrates KIC and KIVA was not reconstituted by TPP + Mg++ alone, but CoASH + NAD could reconstitute the enzyme activity with KIC and KIVA in different degrees. Only two control strains were tested with MEVA as substrate, and these showed in contrast that TPP + Mg++ could partly reconstitute the enzyme activity. In contrast to the relative homogeneity in the reconstitution profiles of normal strains, the five classical and one intermittent MSUD strains showed heterogeneity in cofactor requirements. Complementation analysis using heterokaryons prepared from fibroblasts of four patients with classical MSUD and one patient with intermittent MSUD showed, in contrast to experiments with normal controls, a partial amelioration of the defect in two combinations; it is suggested that the defect in these strains is located at different functional subunits of the multienzyme complex.


Herpesnephropathy.

Herpesnephropathy. Two cases of acute renal insufficiency occurred in association with episodes of severe encephalitis due to herpes simplex type I. The possibility was considered that the renal failure was due to viral infection of the kidneys, and animal experiments were carried out in an attempt to confirm this. Young New Zealand albino rabbits were infected i. v. with HSV type I; the virus antigen was detected in the kidney of 8 of 10 animals, and IgG was found on the GBM in 9 of 19 animals. Viruria was observed in 12 of the 29 infected animals, and electronmicroscopic examination confirmed the presence of immune complexes in the glomeruli.


Oestrogens and endometrial cancer: a point of view. The medical profession has been disturbed and bewildered by a number of recent communications which discussed a possible relationship between contraceptive sequential therapy and the incidence of endometrial cancer and between the postmenopausal application of conjugated oestrogens and endometrial carcinoma. An analysis of these reports is made critically and in detail and supplemented with observations on the risk of carcinoma of the breast. The practical aspects are delineated and a precise guide to postmenopausal oestrogen therapy presented.


Electron spin resonance as a useful technique in the management of Hodgkin's disease. Electron spin resonance spectroscopy has proved a useful and simple technique for the measurement of levels of caeruloplasmin and iron transferrin in whole blood from 50 patients with Hodgkin's disease. Those patients with clinically active disease show higher caeruloplasmin levels and lower iron transferrin levels than those with inactive disease. The results indicate that these tests are good indicators of the state of the disease and that serial measurement of these parameters may help in early prediction of clinical reactivity and in monitoring response to treatment. The combined information from iron transferrin and caeruloplasmin levels appears to be more predictive than that from the erythrocyte sedimentation rate and neutrophil alkaline phosphatase score.


Electron spin resonance measurements of blood caeruloplasmin and iron transferrin levels in patients with non-Hodgkin's lymphoma. Caeruloplasmin and iron transferrin level were measured in blood of patients with non-Hodgkin's lymphoma in different stages of disease activity and compared with erythrocyte sedimentation rate and NAP level in the same samples. It was found that both caeruloplasmin level and sedimentation rate showed a slight increase in mean level in patients with active disease as compared with those in remission, particularly in the group of patients with poorly or undifferentiated diffuse disease. No difference was observed in levels of iron transferrin or NAP. Both caeruloplasmin and sedimentation rate showed occasional abnormal values in patients in remission but in most cases where both were elevated the patients subsequently entered a more active phase of the disease.


The nature of the beta-adrenoreceptor controlling plasma renin activity in man. 1. Seven healthy sodium-replete male volunteer subjects remained supine during and for at least 1 h before the study. Heart rate and blood pressure were recorded continuously, and peripheral venous blood samples were taken every 15 min for determinations of plasma renin activity. 2. All subjects were studied twice: after 3 days of oral practolol (100 mg, three times daily) and after a similar period on placebo. Each study consisted of an intravenous infusion of isoprenaline in graded doses (0-20 microng/min in the placebo phase; 0-16 microng/min in the practolol phase), followed after rest for 2 h by an intravenous infusion of salbutamol (0-20 microng/min after placebo; 0-80 microng/min after practolol). 3. Both salbutamol and isoprenaline produced dose-related increases in systolic blood pressure, heart rate and plasma renin activity and decreases in diastolic pressure. 4. The increases in heart rate and plasma renin activity induced by either agonist were competitively blocked by practolol, as was the fall in diastolic blood pressure induced by isoprenaline; the salbutamol-induced fall of diastolic blood pressure was unaffected by practolol. 5. Comparison of dose ratio--1 estimates confirmed that practolol selectively blocked increases in heart rate and plasma renin activity due to salbutamol; no selective blockade against isoprenaline-induced changes was shown. 6. Selective blockade of salbutamol-induced changes indicate that a beta1-adrenoreceptor mediates changes in plasma renin activity.


Noradrenaline sensitivity of an isolated perfused artery: effects of normal and hypertensive plasma. 1. The noradrenaline sensitivity of the isolated perfused rabbit artery was studied, during perfusion with Krebs solution, normal plasma, hypertensive plasma and a low-density lipoprotein (LDL) extract of normal plasma. 2. There was no difference in noradrenaline sensitivity between arteries perfused with Krebs and normal plasma. 3. Both hypertensive plasma and LDL potentiated the noradrenaline-induced vasoconstriction. 4. This potentiation was associated with increases in both the intrinsic activity of the drug-receptor complex and the drug-receptor affinity. 5. There is a circulating factor in the plasma of hypertensive patients which sensitizes arteries to noradrenaline, and this factor may be LDL.


Isolation of renal membranes that contain kallikrein, angiotensin I-converting enzyme (kininase II) and angiotensinase in the rat. 1. Fractions highly enriched in plasma membrane, endoplasmic reticulum or brush border were prepared from rat kidney cortex. Kallikrein was concentrated in the plasma membrane fraction, but not in the brush border fraction. Angiotensin I-converting enzyme (kininase II) and angiotensinase were localized in the brush border membrane. 2. It is suggested that kallikrein in the urine may originate from plasma membrane distal to the brush border of proximal tubules and the conversion of angiotensin I and the inactivation of bradykinin and angiotensin II may occur on the lumen membrane of the proximal tubular cells.


Functional-morphological correlates of renomedullary interstitial cells. 1. Indomethacin treatment of renomedullary interstitial cells (RIC) causes a marked increase in number and size of their lipid droplets. The anti-hypertensive function is retained. 2. After multiple passages in tissue culture, RIC lose their ability to produce an anti-hypertensive effect in hypertensive animals. Along with this functional loss, lipid droplets and a prominent cisternal system become attenuated or disappear. 3. We conclude that preservation of the lipid granule-cisternal organelle relationship is important in preservation of the anti-hypertensive endocrine function of RIC.


Further unravelling of the causes of ACTH-induced hypertension in the sheep. 1. Acute severe sodium subtraction (20-25% of total exchangeable sodium) before or during treatment with adrenocorticotrophic hormone (ACTH) does little to modify the increase in blood pressure induced by ACTH. 2. Chronic low salt diet, less than 5 mmol/day, abolishes the blood pressure increase, but the response can be restored by increasing the sodium intake to as little as 10 mmol/day. 3. 17alpha,20alpha-Dihydroxyprogesterone infused concurrently with other adrenal steroids will mimic ACTH hypertension and perhaps represents a new class of steroid capable of influencing blood pressure.


Aldosterone regulation in primary aldosteronism: differences between adenoma and bilateral hyperplasia. 1. The diurnal patterns of plasma aldosterone, plasma renin activity (PRA), cortisol and adrenocorticotrophic hormone (ACTH) in the supine and in the upright position have been studied in fourteen patients with primary aldosteronism, five with adenoma and nine with bilateral hyperplasia. Blood samples were drawn at intervals from 6 h to 30 min. 2. Supine patients with an adenoma showed marked diurnal variations of aldosterone, with maximal values at 08.00 hours and minimal values of 18.00 hours and secretory spurts beginning after 02.00 hours. Plasma cortisol paralleled aldosterone, and ACTH seemed to anticipate aldosterone and cortisol variations; PRA remained unchanged. In patients with hyperplasia, aldosterone was significantly lower than in the adenoma group at 08.00 hours, and its decline during the day was less marked; fluctuations rather than secretory episodes were seen. 3. After patients assumed the upright posture, aldosterone remained unchanged or decreased in patients with adenoma, whereas it significantly increased in hyperplasia; PRA remained low, although a slight increment was seen in the latter group. The different response of aldosterone in the two groups was not modified by the administration of propranolol, apparently excluding a renin-dependent mechanism. On the other hand, dexamethasone seemed to affect the response of aldosterone to the upright posture in both groups; in adenoma there was a slight but significant increase, and in hyperplasia the usual rise was partially suppressed. 4. It is concluded that ACTH has a predominant role in regulating aldosterone secretion in primary aldosteronism due to adenoma, whereas its action in bilateral hyperplasia is only permissive.


Selective hypoaldosteronism: a study of steroid biosynthetic pathways under adrenocorticotrophin and angiotensin II infusion. 1. The functional integrity of the adrenal cortex has been tested in a case of selective hypoaldosteronism by adrenocorticotrophin (ACTH) and angiotensin II (AII) infusion. 2. During ACTH infusion a normal functioning zona fasciculata was indicated by the impressive increase of the ACTH-dependent plasma steroids; the aldosterone response was moderate. 3. During AII infusion the plasma aldosterone response was blunted with an unexpected dose-dependent increase in pregnenolone, resulting in abnormal decreasing progesterone/pregnenolone ratios during the infusion, suggesting a slow-down in the conversion of pregnenolone into progesterone. 4. This defect, a probable consequence of chronic renin deficiency on the zona glomerulosa, could be a contributing factor to the hypoaldosteronism.


Cadmium-induced proximal tubular dysfunction in a cadmium-polluted area. Health examinations were performed in 147 people living in a cadmium-polluted area, Kosaka Town, Japan. 33 of 147 residents had some indications of proximal tubular dysfunction, such as renal glucosuria, tubular proteinuria, and aminoaciduria, and 10 of them were diagnosed as having multiple proximal tubular dysfunctions. Detailed examinations revealed that none of the cases had any causal diseases other than chronic cadmium poisoning. Residents' mean cadmium intake and mean urinary cadmium concentration were over 3 times as high as those in control areas. From these findings, renal lesions identified in these residents were concluded as chronic cadmium poisoning induced by environmental cadmium pollution.


Orderly bioelectric phenomena and myelin integrity: an hypothesis. The classical approach to myelin function, in light of current data, is deemed inadequate. Electron microscopic data is used to support the novel hypothesis of interneuronal communication at nodes of Ranvier in the central nervous system of higher organisms. It is hypothesized that myelin plays a role in imposing order hypothesis, use is made of both developmental and degenerative aspects of myelin.


1-alpha-acetylmethadol (LAAM), methadone and morphine abstinence in dependent rats: EEG and behavioral correlates. Adult female Sprague-Dawley rats were prepared with chronic intravenous cannulas and cortical and muscle electrodes for recording electroencephalograms and electromyograms, respectively. They were made physically dependent on morphine by automatic intravenous injections and then trained to lever press in order to self-administer morphine on a FR-20 schedule of reinforcement. Upon stabilization of morphine self-administration, one group continued to self-administer morphine, while two other groups were switched to methadone or 1-alpha-acetylmethadol (LAAM) self-administration for an additional five to ten days. Continuous EEG and EMG recordings were collected. Initially, automatic injections of morphine suppressed rapid eye movement (REM) sleep time, then tolerance developed to this effect. REM sleep time in rats self-administering LAAM, methadone or morphine was within the lower limit of the normal range. Following withdrawal, REM sleep was severely suppressed during the first 24 h with morphine and methadone, but only moderately suppressed with LAAM. Increases in lever pressing during withdrawal from morphine and methadone occurred earlier and were more intense and prolonged than for LAAM. The incidence of head shakes peaked earlier and was higher for morphine and methadone during withdrawal than for LAAM. Irritability scores increased for morphine and methadone during the first day of withdrawal, but did not show any increase until the third day for LAAM. These findings suggest that in dependent rats withdrawal from LAAM is less severe than withdrawal from morphine or methadone.


[Developmental anomalies of the liver and carcinoma of the liver in infants and children (author's transl)]. Several forms of intrahepatic developmental anomalies were found on histological examination in 40 children, among them several siblings and polyovular twins. These anomalies affect in principle all structural elements of the liver. In several cases--some of them complicated by cholangitis-there was cholangiofibrosis and cholangiodysplastic pseudo-cirrhosis. In addition, in six children there developed hepatocellular carcinoma between the ages of six months and twelve years. In the course of the development of hepatic carcinoma obligatory nodular for occur. They are made up of light, glycogen-loaded epithelial cells, which later are transformed into small atypical, basophilic cells. These dark cells have a high mitosis rate, have already escaped from the physiological regulation of proliferation. Presumably the various embryonic disorders of liver structure and carcinoma of the liver are based on the action of the same causative factor, which may be of chemical-toxic origin and transferred across the placenta.


A-cell behavior in organ culture of isolated rat islets: secretion and tissue content of glucagon and insulin. The present study was done to characterize the hormone secretion and content of isolated rat islets in organ culture dependent on the glucose concentration and the number of cultivated islets per culture bottle (10 or 30). For the calculation of the recovery rate the glucagon and insulin breakdown in the culture medium containing 10% calf serum was assayed. The glucagon recovery after incubation for 24 h at 37 degrees C varied from 2 to 92% and that of insulin from 75--105% depending on the serum batches. The cultivated islet responded to 16 mmol glucose by a significant enhancement of insulin release, whereas the glucagon release was not influenced. The glucagon release fell progressively with the duration of cultivation. 2 mmol dibutyryl cyclic AMP augmented insulin and glucagon release in the presence of 5 and 16 mmol glucose and significantly enhanced the glucagon content in the cultivated islets.


Effect of thyroxine and of thyrotropin releasing hormone on cyclic AMP concentration in the anterior pituitary gland in vitro. The effect TRH and thyroxine (T4) in vitro on the content of cAMP in the anterior pituitary gland in rat have been investigated. Thyroxine did not significantly affect this concentration, but TRH produced approximately a two-fold increase in the pituitary cAMP content. Preincubation of the anterior pituitary together with T4--under the conditions when the latter inhibited both proliferative and secretory response of the gland to TRH--did not prevent the elevation of cAMP due to TRH. It is assumed that T4 must have exerted its inhibitory effect outside of the cyclic AMP system.


Modulation of pituitary thyrotropin releasing hormone receptor levels by estrogens and thyroid hormones. The effect of estradiol and thyroid hormone treatment on pituitary TRH binding and TSH and PRL responses to the neurohormone was studied. A significant increase in the number of pituitary TRH binding sites was observed between 2 and 4 days after daily administration of estradiol benzoate with a plateau at 300% of control being reached at 7 days. Plasma PRL levels showed a similar early pattern of response. In animals rendered hypothyroid by a 2-month treatment with propylthiouracil or 1 month after surgical thyroidectomy, the level of pituitary TRH receptors was increased approximately 2-fold, this elevation being completely reversed by treatment with thyroid hormone. Estradiol-17beta administered with L-thyroxine partially reversed the inhibitory effect of thyroid hormone on TRH receptor levels in hypothyroid animals. The antagonism between estrogens and thyroid hormone is also apparent on the TSH response to TRH since estrogen administration can reverse the marked inhibition by thyroxine of the TSH response to TRH either partially or completely in intact and hypothyroid animals, respectively. The PRL response to TRH is 55 and 40% inhibited in hypothyroid and intact rats, respectively, by thyroid hormone when combined with estrogen treatment. The present data clearly show that estrogens and thyroid hormones can affect TSH and PRL secretion, the effect of estrogens being predominantly on PRL secretion while thyroid hormone affects mainly TSH. The close correlation observed between the level of TRH receptors and PRL and TSH responses to TRH suggests that estrogens and, to a lesser extent, thyroid hormones, exert their action by modulation of the level of receptors for the neurohormone in both thyrotrophs and mammotrophs.


Changes of pituitary thyrotropin releasing hormone (TRH) receptor level and prolactin response to TRH during the rat estrous cycle. The plasma PRL and TSH responses to TRH injected iv at different stages of the estrous cycle in normal rats under Surital anesthesia were maximal during the afternoon of proestrus and morning of estrus and lowest on diestrus I. As calculated from the areas under the plasma response curves, a 10-fold difference was found between the maximal and minimal PRL responses while a 2-fold difference was measured for TSH. The plasma PRL and TSH responses to TRH showed a correlation with the binding of [3H]TRH to anterior pituitary gland, a 3-fold difference being observed between the minimal binding measured on the morning of diestrus II and the maximal value found on the evening of proestrus. Contrary to findings with LHRH and LH, repeated injections of a small dose  (10 ng) of TRH in the afternoon of proestrus abolished PRL and TSH responses to subsequent injection of the neurohormone.


Triiodothyronine receptors during maturation. Capacity of rat liver nuclear triiodothyronine (T3) binding protein (NTBP) for T3 more than quadruples from birth through 50-120 days, whether related to DNA or weight of tissue. Ka for T3 doubles during this interval. T3 content of nuclei nearly parallels capacity. The proportion of receptors occupied by T3 is 0.3 at birth, falling to .16 in old rats. In contrast to these changes in liver content of T3-receptor complex, mitochondrial alpha-glycerophosphate dehydrogenase activity changes relatively less during maturation. Administration of estrogen, testosterone, and dexamethasone to immature female rats did not alter NTBP capacity of affinity.


Reevaluation of the role of cyclic adenosine 3',5'-monophosphate and protein kinase in the stimulation of steroidogenesis by luteinizing hormone in bovine corpus luteum slices. Incubation of bovine corpus luteum slices at 37 C with luteinizing hormone showed that 10.0 microng LH/ml caused a maximum rise in tissue cyclic AMP content within 15 min. Slices incubated with 1.0 or 0.1 microng LH/ml showed a much more gradual accumulation of this nucleotide. In the absence of added LH, a marked decline in the amount of cyclic AMP was observed during the first 60 min. The possible role of cyclic AMP in the action of LH was reexamined by studying the dose-response effect of LH on the stimulation of progesterone synthesis, cyclic AMP-dependent protein kinase activation, and cyclic AMP accumulation. After a 2-h incubatin, the results showed that the dose required to elicit a minimal significant stimulation of steroidogenesis was 0.01 microng/ml LH. At this and higher concentrations of LH, a concomitant stimulation of protein kinase activity and progesterone synthesis was also consistently observed. However, significant accumulation of cyclic AMP became consistently detectable only at 0.1 microng/ml LH. This report is the first to show a positive correlation between the activation of cyclic AMP-dependent protein kinase and the stimulation of steroidogenesis in the corpus luteum at the same minimal effective level of LH. These results indicate that cyclic AMP and the cyclic AMP-dependent protein kinase probably play important intermediary roles in the stimulation of steroidogenesis by LH in the bovine corpus luteum.


Perchlorate ion enhances mouse thyroid responsiveness to thyrotropin, human chorionic gonadotropin and long acting thyroid stimulator. Perchlorate treatment of mice increased by 1.5-2-fold the thyroid secretory response to TSH, hCG and LATS, in the McKenzie bioassay. Perchlorate alone did not increase basal plasma radioactivity. Perchlorate augmentation of the secretory response index was roughly proportional to the level of stimulation; it was similar for all three stimulators despite their different time courses of action which were unaltered by perchlorate; it was the same whether perchlorate administration preceded, coincided with or shortly followed injection of the stimulator, a finding in keeping with the slow clearance of this ion. The perchlorate effect was dose-related, although within a narrow range (6.25-12.5 microng/mouse). Near-maximal per chlorate effect was obtained with a dose (12.5 microng) which, when tested in different experimental conditions (MMI-blocked thyroid), discharged 80% of intrathyroidal radioiodide. Perchlorate exerted its augmenting effect by enhancing thyroid secretion: it increased plasma radioiodothyronines and radioiodide concentrations without decreaseing the blood disappearance rates of iodide and iodothyronines. The potentiating effect of perchlorate probably takes place at a step prior to cyclic AMP action since it did not affect dbcAMP-stimulated secretion. The perchlorate effect may be indirect, through mobilization of minute amounts of intrathyroidal iodide.


Female steroid hormones and lipoprotein synthesis in the cockerel: effects of progesterone and nafoxidine on the estrogenic stimulation of very low density lipoproteins (VLDL) synthesis. Since progesterone is known to modulate the effects of estradiol in a number of organ systems, we have studied the effects of the two hormones on very low density lipoproteins (VLDL) synthesis in the cockerel. We have also examined the effect of an estrogen antagonist, nafoxidine-HCl, on the process. When three week-old cockerels were treated with a single injection of estradiol (1 mg) or estradiol (1 mg) + progesterone (2 mg), plasma VLDL increased within 5 h, reached a peak at 48 h and returned to baseline values at 68 h. There was no significant difference between estradiol or estradiol + progesterone treated animals. Liver slices were prepared from similarly treated animals and incubated in vitro with L-[3H]lysine for 2 h. Following homogenization and centrifugation at 105,000 X g, newly synthesized VLDL were quantitiated in the supernatant fluid by immunoprecipitation with a monospecific antibody. Radioactivity incorporated into VLDL was found to increase from low baseline levels to a peak at about 17 h after treatment. There was, again, no significant difference between estradiol and estradiol + progesterone-treated animals. Estrogen priming or progesterone pretreatment also did not significantly alter the VLDL biosynthetic response in liver slices to estradiol alone or in combination with progesterone. When the estrogen-antagonist nafoxidine-HCl (5 mg) was administered simultaneously with estradiol (1 mg), it totally inhibited the VLDL biosynthetic response in liver slices in vitro. The interaction of estradio, progesterone and nafoxidine-HCl on the hepatic synthesis of VLDL should serve as a valuable model for the study of VLDL synthesis and its regulation as well as the mode of action of the sex steroid hormones in the liver.


Relationship of biochemical and morphological changes in rat thyroid and proton spin-relaxation of the tissue water. The effect was studied of biochemical and morphological changes induced by antithyroid substances (PTU, C10(-4)) on proton spin-relaxation properties of rat thyroid gland. It was found that thyroid stimulated by PTU (0.05%) or C10(-4) (1.0%) exhibit marked morphological changes (hyperplasia and epithelial hypertrophy) with alteration of the soluble iodoprotein pattern (content and composition.). Both relaxation times spin-lattice (T1) and spin-spin (T2) were increasing with the lenght of treatment with antithyroid drugs. Reversibility of the process was noted in accordance with biochemical and morphological data. The relaxation rate (formula: see text) for thyroid tissue water was in positive correlation with the suluble protein concentration and particularly with the TG content in the gland. There was no difference in relaxation times between normal thyroid and gland of rats treated chronically with excess iodide. The observed difference in T1 between normal glands and glands of PTU,-C10(-4)--treated rats was comparable with that found in cases of human thyroid cancer. This finding is of importance when the diagnostic potential of NMR in the detection of malignancy is considered. In conclusion, a strong correlation was found between microstructural and biochemical changes of the thyroid gland and proton magnetic relaxation of tissue water. The striking difference between the proton spin-relaxation times in normal and in goiter thyroid glands of rats suggests that pulsed NMR spectroscopy could be a method for evaluation of some disturbances in thyroid gland.


Fine structure of adrenal cortex in ectopic ACTH syndrome. A 49-year-old man with pancreatic adenocarcinoma, suggestive of islet cell origin, showed clinical and biochemical features of ectopic ACTH syndrome and underwent bilateral adrenalectomy. Light microscopy revealed adrenocortical compact cell hyperplasia and lipid depletion. The zona glomerulosa was detected in small foci and fasciculata cells extended up to the capsule. Electron microscopy disclosed enlargement of adrenocortical cells, massive SER accumulation, RER increase, lipid depletion, prominence of the Golgi apparatus and development of complex interdigitations between closely apposed cell membranes. These changes were attributed to the stimulative effect of ACTH discharged from the non-pituitary tumor. Mitochondria exhibited enlargement, pleomorphism and cavitation. Most of the adrenocortical cells contained mitochondria with vesicular cristae--a characteristic feature of fasciculata cells. In a few cells, under the capsule, some mitochondria possessed lamellar cristae. Gradual transformation to the vesicular type was, however, apparent indicating that mitochondria are not rigidly constant structures.


Post-EEG seizure depression of human limbic neurons is not determined by their response to probable hypoxia. A subclinical EEG seizure occurred in each of 2 patients during a test that repeatedly evoked probable mild cerebral hypoxia. The firing rate of the neurons was unaffected until the EEG seizure had been ongoing for over a minute. After the EEG seizures, all neurons were profoundly depressed regardless of whether their responses to hypoxia had been an increase, decrease, or no change in firing rate. These observations indicate that the mechanism responsible for post-EEG seizure depression of neuronal firing is probably not cerebral hypoxia.


Prostaglandin E2 production in 3T3 cells transformed by polyoma virus raises the intracellular adenosine 3':5'-monophosphate levels. High cellular adenosine 3':5'-monophosphate (cyclic AMP) were found in a polyoma-virus-transformed 3T3 fibroblast line, which produces comparatively high prostaglandin E2 concentrations. Prostaglandin E2 and cyclic AMP increased with time during 6 days of growth. Both effects were prevented by three different cyclo-oxygenase inhibitors. Addition of prostaglandin E2, at a concentration which would been synthesized in the absence of inhibitor, reversed the effect of indomethacin, one of the cyclo-oxygenase inhibitors, on cyclic AMP. It is concluded that endogenous prostaglandin E2 production in these transformed cells influences their cellular cyclic AMP levels.


Prostaglandin production by normal and transformed 3T3 fibroblasts in cell culture. Prostaglandin E2 and prostaglandin F2alpha were quantitatively determined in culture media from Balb/c 3T3 fibroblasts and two virus-transformed derivatives of these cells. Regular and (simian virus 40)-transformed 3T3 cells produced low and almost identical prostaglandin E2 concentrations. The levels were maximal (10 ng/ml) 24 h after planting and decreased during the next 96 h to 2.5 ng/ml. In contrast, polyoma-virus-transformed 3T3 cells produced prostaglandin E2 continuously for 120 h, giving a final concentration of 270 ng/ml. Fresh medium supplemented with calf serum reinitiated prostaglandin production in confluent 3T3 cells. Maximal prostaglandin E2 concentrations were obtained between 6 and 18 h after medium change and were proportional to the serum concentration of the medium. Indomethacin (10(-6) M) completely inhibited prostaglandin E2 production but not prostaglandin F2alpha production by these cells.


Phospholipid binding and self-association of the major apoprotein of human and baboon high-density lipoproteins. The purpose of this study was to establish a relationship between self-association and phospholipid binding of the human and the baboon apoA-I protein. The enthalpy changes on binding dimyristoyl lecithin and lysolecithin to either the human or the baboon native apoA-I protein were measured in a microcalorimeter. An endothermal process, most pronounced for the human apoprotein, was observed at low phospholipid levels. At higher phospholipid to protein ratios the binding was exothermal. Gel filtration experiments on Sephadex G-200 showed that the native apoprotein of both species consists of dimers and tetramers. The baboon native apoA-I protein contained a higher amount of dimers. After preincubation of the apoA-I protein with lysolecithin, the enthalpy changes measured on subsequent binding of dimyristoyl lecithin were shifted towards more exothermal values compared to the curve for the native apoprotein. The amplitude of this shift corresponds to that of the endothermal process observed on binding dimyristoyl lecithin to the native apoprotein. This process was attributed to a phospholipid-induced disaggregation of the apoA-I protein. Gel filtration data showed a decreased extent of aggregation in the apoA-I protein preincubated with lysolecithin. This sample consisted exclusively of dimers. Ultracentrifugal flotation of the complexes formed between the apoA-I protein, and respectively dimyristoyl lecithin and sphingomyelin indicated that preincubation with lysolecithin increased the extent of complex formation. These results suggest that the dimeric form of the apoA-I protein possesses the highest affinity for phospholipids. Any dissociation of higher polymers enhances the phospholipid-binding capacity of the human and the baboon apoA-I protein.


Thyrotropin binding to and adenylate cyclase activity of porcine thyroid plasma membranes. Plasma membranes have been purified from porcine thyroid gland homogenate by discontinuous sucrose gradient centrifugation. The preparations contained specific binding sites for thyrotropin but not for luteinizing hormone or the beta subunits of thyrotropin and luteinizing hormone. Optimum conditions of 125I-labeled thyrotropin binding were pH 6.0-6.5 and 37 degrees C. Thyrotropin binding was reduced by divalent (Ca2+, Mg2+) and monovalent cations (Na+, K+, Li+), 50% inhibition being obtained at 10 mM and 50 mM respectively. Displacement curves of 125I-labeled bovine or porcine thyrotropin by the unlabeled hormone from three species was in the order of increasing concentrations (bovine greater than porcine greater than human) which is the order of decreasing biological activity of these hormone preparations in the assay in vivo in the mouse. The validity of the results was established by controlling that porcine membranes bound the native and the 125I-labeled hormones with equal affinity. A single type of high-affinity (Kd = 0.28 nM) binding sites was detected for bovine and porcine thyrotropins. In contrast, porcine plasma membranes bound human thyrotropin with a lower affinity (Kd = 70 nM). A good correlation was found at equilibrium and in the conditions of the cyclase assay, between receptor occupancy and adenylate cyclase activation for the three hormones.


UDP-apiose/UDP-xylose synthase. Subunit composition and binding studies. The UDP-apiose/UDP-xylose synthase from cell suspension cultures of parsley has been purified 1400-fold by an improved method. The ratio of apiose to xylose formed from UDP-D-glucuronic acid (UDP-GlcUA) remained constant throughout the purification procedure. Dodecylsulfate-gel electrophoresis and sedimentation equilibrium measurements showed that this enzyme preparation is  composed of two proteins with molecular weights of 65000 and 86000. The two proteins which are present in a molar ratio of about 1:0.7 to 1:0.9 could not be separated by ammonium sulfate fractionation, chromatography on DEAE-cellulose at different pH-values, and on omega-aminoalkyl-Sepharose, and by gel filtration on Acrylex P-100. Each protein is composed of two apparently identical subunits. The presence of only two different subunits was confirmed by end group analysis in which glycine was found as N-terminal amino acid for the larger and lysine for the smaller protein. Crosslinking with dimethylsuberimidate gave dimers of the identical subunits but no hybrids. Separation of the two proteins was achieved on DEAE-cellulose in the presence of urea. After dialysis only the 86000-Mr protein showed enzyme activity with no significant change in the apiose/xylose ratio. However, in the absence of the 65000-Mr protein enzyme stability was decreased drastically. By equilibrium dialysis it was found that 0.5 mol UDP-GlcUA are bound per mole of 86000-Mr protein. NAD+ alone was not bound, but in the presence of UDP it was also bound in a ratio of 0.5 mol/mol catalytic protein. Experiments in which sodium borohydride was added to the enzyme incubation gave no indication that the 4-keto intermediate is bound as a Schiff base to the enzyme. Also no evidence for epimerization at C-3 of the 4-ulose intermediate prior to ring contraction to apiose was found.


Determination of the solution conformation of adenosein 2':3'-monophosphate by nuclear magnetic resonance with lanthanide probes. The aqueous solution conformation of adenosine 2':3'-monophosphate at pH 2.5 has been determined by a nuclear magnetic resonance method utilizing lanthanide ions as shift and relaxation probes. The ribose conformation is best described as a rapid equilibrium of 2'-endo(3'-exo) and 3'-endo(2'-exo) conformations in a ratio of approximately 2 to 1. The orientation of the base relative to ribose is restricted to a narrow range about chiCN=-70 degrees.


Role of divalent ions in folding of tRNA. The native structure of tRNA is not achieved in low salt (4.5 mM Na+, 25 degrees C), but can be restored by addition of divalent ions. We have explored the structure of the central region in Escherichia coli tRNAfMet by absorption and emission spectroscopy of 4-thiouracil, and the structure of the anticodon loop in yeast tRNAPhe by fluorescence of the 'Y' base, versus the number of manganese ions bound to tRNA, which was derived from electron spin resonance. The fluorescence of the reduced 8-13 photoproduct (in which 4-thiouracil at position 8 is crosslinked to cytosine at position 13) was also analysed. In low salt (e.g. 4.5 mM Na+), the region of 4-thiouracil is affected strongly as the first eight Mn2+ bind to tRNA, whereas the fluorescence of the 'Y' base is affected only after four Mn2+ are bound. Considering the structural similarities of the two tRNAs, this suggests that the reorganisation brought about by divalent ions starts in the central region, the anticodon loop being affected later. The binding of divalent ions to each region starts together with its restructuration. Monovalent ions can substitute for divalent ions in this process, a 15 mM sodium concentration being equivalent to the binding of the first five Mn2+. If divalent ions are then added, even the first ones distribute themselves between both the central and the anticodon region. Alternatively, the renaturation may be achieved by monovalent ions only, implying that no sites exist whose occupancy by divalent ions is crucial for the native structure. These observations suggest that the  role and means of divalent ion binding to tRNA are largely explainable in terms of a simple maganese-phosphate binding supplemented by electrostatic interaction with distant phosphates.


Stimulatory and inhibitory effects of dimethyl sulfoxide and ethylene glycol on ATPase activity and calcium transport of sarcoplasmic membranes. 1. The effect of dimethyl sulfoxide (Me2SO) and ethylene glycol on two different preparations of the sarcoplasmic reticulum, i.e. native membranes and membranes whose phospholipids were hydrolyzed by phospholipase A, were investigated using ATP and p-nitrophenylphosphate as substrates. 2. Me2SO and ethylene glycol inhibit both calcium-dependent ATP hydrolysis and ATP-supported calcium transport by native vesicles. 3. In contrast, calcium-dependent p-nitrophenylphosphatase activity as well as p-nitrophenyl-phosphate-supported calcium transport are activated by both agents at concentrations lower than 30% (v/v). 4. Me2SO strongly stimulates p-nitrophenylphosphate activity of vesicles treated with phospholipase A, but has relatively little effect on p-nitrophenylphosphatase activity of native vesicles. 5. Up to a concentration of approximately 40% Me2SO (v/v) the inhibiting effect on the calcium-dependent ATPase is fully reversible, but only partially reversible on calcium transport. 6. In the concentration range where Me2SO inhibits ATP hydrolysis and calcium transport, it does not affect ATP binding to the membranes nor calcium-dependent formation of phospho-protein. 7. The rate of dephosphorylation as well as the rate of Pi exchange between ATP and ADP are markedly reduced by the presence of 30% Me2SO (v/v). 8. While Me2SO inhibits passive calcium efflux, ethylene glycol produces a considerable activation. 9. ADP-dependent calcium efflux and ATP synthesis are activated by 15% Me2SO (v/v). Ethylene glycol reduces both activities. 10. The results suggest that the respective substrate-enzyme complexes are differently affected by the agents, resulting either in inhibition or stimulation


Juvenile familial hypertriglyceridemia and growth retardation. Clinical and biochemical observations in three siblings. Familial hypertriglyceridemia or hyperlipoproteinemia type I was detected in three siblings aged 6, 11, and 14 of an otherwise normal Turkish family of 10 members. Initial values ranged from 1780 to 3750 mg/100 ml triglycerides in the milky white serum; cholesterol was normal. Lipoprotein pattern on agarose and acrylamide gel revealed a heavy band of chylomicrons and missing HDL; post-heparin lipolytic activity was decreased to about 30% of normal. Chylomicronemia could be induced by a fat-rich (50% of total calories) diet, but not by carbohydrates. On a low fat diet (5%) during hospitalization chylomicrons disappeared, and triglycerides decreased to about 450 mg/100 ml. Phenocopies of hypertriglyceridemia could be excluded. All three patients were the only members of the family who were small, below the third percentile. Their bone age was retarded from 18 to 30 months. There was no indication for an endocrine cause of the growth retardation: four different stimulation tests revealed normal growth hormone response, thyroid and adrenal functions were not impaired; sexual development was normal. Increased glucose assimilation was observed during intravenous and oral glucose load. Peak serum insulin response was above normal during stimulation tests. The possible etiologic role of hypertriglyceridemia in this concomitant growth retardation is discussed.


Gamma-aminobutyric acid (GABA) and sleep. The influence of di-n-propylacetic acid on sleep in man. The effects of di-n-propylacetic acid (DPA) on sleep, for some years used as an anticonvulsive drug, has been investigated in 11 healthy volunteers using all-night sleep EEG recordings. DPA acts by an enhancement of the gamma-aminobutyric acid (GABA) level of the brain. Its influence on sleep seemed to be of interest on account of the metabolic relationship of GABA to other short chain fatty acids. After short-term application only a shortening of the time to fall asleep and of the waking time could be found, whereas under long-term administration a decrease in deep synchronous sleep could be observed. In contrast to the results known from animal studies no marked influence on REM sleep was observed. The action of DPA on sleep is similar to that of diphenylhydantoin. No so-called matitudinal 'hangover' could be revelaed in either drug;


Controlled clinical investigation of trimeprazine as a sleep-inducer in normal subjects. The effects of acute introduction and withdrawal of trimeprazine (Vallergan), an antihistaminic phenothiazine derivative with known sedative effects, were investigated by a single blind polygraphic study in 8 healthy volunteers. A baseline placebo was given on Nights 1-3, followed either by 10 mg or 20 mg of the drug on Nights 4-6, and then withdrawal effects were recorded on Nights 7-9. The subjective effects of the drug and EEG sleep variables were determined on these nights. The results showed that the drug might have had dose related effects. REM-sleep was increased by 10 mg doses, which also caused an increase in SW sleep. There was no significant change after the 20 mg dose. Significant REM rebound was not observed after withdrawal of either dose. These characteristics may be of value in the treatment of certain aspects of sleep disturbances.


Effects of several catecholamines on sympathetic transmission to the myocardium: role of presynaptic alpha-adrenoceptors. Effects of several alpha-adrenoceptor agonists on presynaptic alpha-adrenoceptors were evaluated by studying chronotropic responses to cardioaccelerator nerve stimulation in anesthetized dogs. I.v. infusions of norepinephrine (NE), methylnorepinephrine (MNE), epinephrine (E) or phenylephrine (PHE) in desipramine-treated dogs caused significant attenuation of nerve stimulation responses. The inhibitory influence of all these agents could be prevented by prior treatment with phentolamine, but not with haloperidol. Comparisons of relative pressor and presynaptic inhibitory actions of these compounds revealed that in equipressor doses, MNE caused a significantly greater attenuation of nerve stimmulation responses than NE, while PHE had similar pressor and presynaptic inhibitory activity to that of NE. The inhibition of chronotropic responses to nerve stimulation observed following E was significantly greater compared to its relative pressor effect. These results demonstrate the existence of a presynaptic alpha-adrenoceptor mechanism modulating cardiac rate in intact dogs and indicate that the false transmitter MNE may be more potent than NE in imparing neuronal transmission by an action on presynaptic alpha-adrenoceptors.


Dopamine receptors in the femoral vascular bed of the dog as mediators of a vasodilator and sympathoinhibitory effect. ET 495 and apomorphine, injected in small doses (0.1--1 microng/kg) into the femoral artery, induced a dose-dependent increase in femoral blood flow. This dilator effect was abolished by section of the ipsilateral femoral nerve and sciatic nerve, transection of the spinal cord, alpha-adrenoceptor blockade, ganglionic blockade and guanethidine. In addition, the increase in blood flow was inhibited by intravenous administration of haloperiol (2 mg/kg i.v.) or pimozide (2 mg/kg i.v.) and by injection of small doses (10--50 microng/kg) of these drugs into the femoral artery. It was concluded that a dopaminergic component located in the femoral vascular bed of the dog may be involved in the local vasodilator and sympathoinhibitory effect of apomorphine and ET 495.


The mechanism of action of nicotine on vascular adrenergic neuroeffector transmission. The aim of this study was to determine the site and mechanism of action of nicotine on sympathetic neuroeffector transmission in the isolated pulmonary artery of the rabbit. Nicotine and cocaine potentiated the constrictor response elicited by electrical-field stimulation of postganglionic adrenergic neurones. The potentiation was reversible and in the case of nicotine, no tachyphylaxis developed. The nicotine-induced potentiation was characterized by a rapid onset and an initial, transitory peak, while the enhancement caused by cocaine progressed more slowly and was monophasic. Hexamethonium and (+)-tubocurarine prevented the potentiation caused by nicotine. Nicotine did not prevent the adrenergic neurone blocking effect of bretylium on the response to field stimulation. Nicotine increased the stimulation-induced outflow of tritium from pulmonary artery preloaded with 3H-(--)-noradrenaline. Contractions of the artery elicited by tyramine were enhanced by pargyline, unaltered by nicotine and blocked by cocaine. Nicotine did not alter the concentration--response curve of exogenous (--)-noradrenaline while cocaine moved it to the left. The accumulation of 3H-(--)-noradrenaline by rabbit isolated aorta was not altered by nicotine, hexamethonium and (+)-tubocurarine. The accumulation of 3H-nicotine by the aorta was much lower than that seen with 3H-(--)-noradrenaline. The disposition of the 3H-nicotine accumulation into adventitia and media was concentration-independent. These results suggest (1) that nicotine potentiates the neurogenic vasoconstriction response in part by increasing the stimulation-induced release of transmitter from adrenergic neurone terminals; (2) that the site of the nicotinic receptors mediating this action is located on the outer surface of the neurones; and (3) that the potentiation is not due to blockade of noradrenaline re-uptake.


The inhibitory effect of cadmium on neuromuscular transmission in the rat. Cadmium (0.125-1 mM) was found to inhibit the isometric response of the isolated rat hemidiaphragm during indirect stimulation, but not during direct stimulation. This effect of cadmium (1 mM) was completely reversed by ethyleneglycol bis-(aminoethyl)-N,N,N',N'-tetra-acetic acid (2 mM) or by L-cysteine (2 mM) but only partially by increased calcium. Cadmium (10 micronM) significantly reduced the quantal release of transmitter in the isolated phrenic diaphragm and a concentration of 0.1 mM frequently caused a complete failure of the endplate response after 30 min. The effect of cadmium on neuromuscular transmission could not be readily reversed by washing with cadmium-free solution. Miniature endplate potential frequency and amplitude were not significantly affected by cadmium (0.1 or 0.5 mM). The results suggest that the effect of cadmium on the isolated phrenic nerve-diaphragm is due largely to inhibition of calcium function at presynaptic nerve terminals.


Potentiation by crude kallikrein of the myotropic effect of angiotensin I in the isolated rabbit aortic strip. Crude kallikrein (Padutin), but not pure kallikrein, when preincubated with angiotensin I caused a potentiation of the myotropic effect of decapeptide on the isolated continuously superfused rabbit aortic strip. Addition of converting enzyme inhibitor, SQ 20881, to the medium inhibited this potentiation. The potentiation by crude kallikrein of the myotropic effect of angiotensin I is probably due to the conversion of decapeptide to octapeptide angiotensin II. This study indicates that Padutin is not a pure kallikrein preparation and probably contains a kininase fraction which causes the conversion of angiotensin I.


The effect of levamisole on phosphodiesterase activity. Phosphodiesterase activity of mouse liver homogenates was estimated in presence and absence of levamisole. The enzyme activity was 1394 and 1399 nmoles/mg protein/30 min respectively. Our data show that levamisole does not affect the phosphodiesterase activity.


Dexamethasone suppression of ovulation in PMS-treated immature rats. A single injection of 2.0 mg/kg dexamethasone (DXM) administered at 51 h after pregnant mare serum gonadatropin (PMS) treatment inhibited both ovulation and luteinization. S.c. injection of human chorionic gonadotropin (HGG) caused ovulation and luteinization in DXM-PMS-treated rats, whereas treatment with ACTH failed to overcome the DXM inhibitory effect. These findings are interpreted to indicate that DXM inhibits ovulation through a mechanism which might involve the central nervous system.


[Antiviral agents. XIX. Amantadine derivatives and several others]. The synthesis and pharmacological activity of several adamantane derivatives and analogs of tilorone are described as part of a systematic research on potential antiviral compounds and/or interferon inducers.


Evolution of neurohypophyseal hormones and their receptors. Nine active neurohypophyseal principles have been isolated and identified among the vertebrates. Arginine-vasotocin is the most ubiquitous, occurring in pituitary glands from representatives of all the major vertebrate groups. There is much more variation in structure among the principles that resemble oxytocin. The manner in which these evolved remains unclear. Arginine-vasotocin stimulates smooth muscles from a wide variety of vertebrate species. It can stimulate contraction of oviducts from many jawed fishes and tetrapods. The oxytocin-like peptides are usually less active in this respect. Among adult mammals arginine-vasotocin is replaced by arginine-vasopressin which has much less oxytocin activity. Thus, although arginine-vasotocin may both stimulate oviducts and cause water retention in nonmammalian tetrapods, oxytocic and antidiuretic functions can be regulated independently by oxytocin and vasopressin in mammals. Arginine-vasotocin elicits vasoconstrictor responses in even the most primitive vertebrates. These may be systemic or regional. Their distribution may determine whether arginine-vasotocin acts as a diuretic or an antidiuretic agent. It is possible that the most primitive neurohypophyseal functions were related to cardiovascular regulation and that the neurohypophysis acquired its osmoregulatory functions later in vertebrate evolution.


Characterization of oxytocin receptors in the uterus and mammary gland. High affinity binding sites for 3[H] oxytocin have been demonstrated in particulate fractions from rat uterus and oviduct, myometrium from the sow, ewe and human, ewe endometrium, and mammary gland from the lactating rat. The binding activity has been localized to enriched plasma membrane fractions from the rat uterus and mammary gland; cells isolated from the mammary gland also bind oxytocin. The apparent dissociation constant (Kd) for the interaction of oxytocin with its binding sites in a variety of tissue preparations is in the nanomolar range. The concentration of oxytocin eliciting half-maximal contraction of the rat isolated uterus corresponds to the apparent Kd of oxytocin interaction with uterine particulate fractions. Binding is specific with respect to the target tissue or cell, as well as to the ligand. The affinity of binding sites for oxytocin analogues corresponds generally to their potencies as agonists or antagonists. Factors that affect the binding of oxytocin affect the biological response in the same way. For example, certain divalent metal ions, which increase oxytocin binding activity, enhance the sensitivity of the contractile response of the uterus and mammary gland to oxytocin. Estrogen administration, which increases the uterine binding of oxytocin, increases the sensitivity of the myometrium to oxytocin. The myometrium binds the most oxytocin at estrus and is most sensitive to oxtocin at that time. The dgree of stimulation by oxytocin of prostaglandin F2alpha synthesis by ewe endometrium is paralleled by an increased concentration of oxytocin binding sites. The marked increase in sensitivity to oxytocin of the rat uterus occurring on the day of parturition also is reflected by the amount of oxytocin bound by the uterus. Because of the many correlations between oxytocin binding and bioactivity, it appears that oxytocin binding sites on the plasma membrane of target cells constitute the recognition part of oxytocin receptors.


Cyclic nucleotides in the cellular action of neurohypophyseal hormones. Cyclic 3',5'-nucleotides play an important role in the action of neurohypophyseal hormones on peripheral tissues. All available evidence indicates that cyclic AMP serves as an intracellular mediator in the regulatory action of neurohypophyseal hormones on transport of fluids and solutes across both mammalian and nonmammalian epithelial membranes. There is a close association among binding of neurohypophyseal hormones on membrane, stimulation of cyclic AMP generation, and the functional response. On the other hand, neurohypophyseal hormones have no similar effect on cyclic AMP metabolism in contractile tissues such as smooth muscle. It appears likely that neurohypophyseal hormones stimulate primarily generation of cyclic GMP in contractile tissues, and the increase in cyclic GMP levels may be associated with the contractile response. While the role of cyclic AMP in neurohypophyseal hormone effects in epithelia is firmly established, the possible role of cyclic GMP in contractile responses is largely hypothetical at the present time.


Indentification of sites in oxytocin involved in uterine receptor recognition and activation. Following a brief review of the preferred three-dimensional structured proposed for oxytocin in dimethylsulfoxide and in aqueous medium, the "cooperative model" for the biologically active conformation of oxytocin is discussed. An approach to conformation-activity analysis is described. The importance of determining the peptide backbone conformation and the functional contribution of peptide backbone and amino acid side chains to hormone receptor interactions are analyzed. Sites are proposed that are thought to be involved in the binding process of oxytocin to the smooth muscle receptor in rat uterus, as well as sites that are thought to be responsible for receptor activation.


The biological effects of polybrominated biphenyls in avian species. Polybrominated biphenyl (PBB), structurally similar to polychlorinated biphenyl (PCB), affects feed intake, egg production, and hatchability of fertile eggs in chickens and quail. Subcutaneous edema of the neck and shoulder are common abnormalities of hatched chicks from PBB-fed dams. No characteristic teratogenicity is evident. Offspring hatched are less viable during their first 3 weeks of life. PBB does not produce thinner egg shells or lower egg weights. The ratio of PBB egg content to dietary level is 1.3-1.5:1.0. The biological half-life of PBB in eggs is calculated to be 17 days. Tissue residues are higher in males than females because egg production is a major excretory route for PBB. Polybrominated biphenyl feeding causes tissue changes in size, structure and/or function in the liver, thyroid, testes, comb, heart, spleen, bursa of Fabricus, and blood. Liver enlargement, porphyria and microsomal enzyme induction results from PBB administration. The spleen, bursa and comb are reduced in size, whereas the thyroid increases after PBB feeding. A characteristic of PBB and PCB toxicity is hydropericardium and general edema.


[Mechanisms of participation of the sensomotor cortex in controlling movements]. Neuromorphological, neurophysiological and biomechanical studies in cats revealed cortical structural neuronal modules and functional units of different grades of complexity. Single systemic principle was determined for the sensorimotor cortex activity: structurally organized neuronal modules, having distributed outputs, can be involved in different functional units for purposeful movements. Specific lemniscal signals concerning the movements are the factor which filters inflow of teleceptive information to the neuronal modules and determines exact torographical relations of cortical motor outputs.


[Sensitivity to adrenocorticotropic hormone during adaptation to increased muscle activity]. Systematical muscle exercises increased sensitivity of adrenals to ACTH in adult albino rats, while fatigue decreased it. The sensitivity of adenylatecyclase of fat cells to ACTH does not change during the adaptation of organism to increased muscular activity. Activation of adenylate cyclase of fat cells with epinephrine is abolished by sympatholytin which, however, has no effect on the ACTH activation. These findings confirm that the adenylate cyclase receptors for epinephrine and for ACTH are different.


The effect of insulin treatment on the metabolic response to glucagon in diabetes. The metabolic response to intravenous glucagon was studied in insulin requiring diabetes before, 1 week and 1 month after treatment with insulin. The rise in plasma 3-hydroxybutyrate following glucagon in the untreated state was converted to a fall after 1 month on insulin. It is suggested that this response could provide a measure of the biological effectiveness of circulating insulin. There was no change in plasma cyclic AMP response to glucagon after treatment with insulin.


Androgen stimulation of prolactin receptors in rat prostate. Specific receptors for iodine-labelled human prolactin ([125I]hPrl) are present in membrane preparations of the rat ventral prostate. The binding is saturable with an apparent association constant (Ka) of 2.2 X 10(9) M-1 and a binding capacity of about 1 pmol/100mg prostatic tissue. The binding of [125I]hPrl is inhibited by hPrl, ovine Prl (otprl) and human growth hormone, but not by ovine FSH or LH. Serum from rats having Prl-producing pituitary tumors caused a displacement of the [125I]hPrl from the receptors, and the displacement curve was parallel with that of the hPrl standard. Treatment of immature rats with varying doses of dihydrotestosterone propionate (10-5000 microng) causes a dose-dependent stimulation of Prl receptors calculated both as binding sites per mg of membrane protein and as binding sites per prostate. Androgen stimulation of prostatic Prl receptors increases the tissue sensitivity for circulating Prl and may be one reason for the known increases in endogenous cAMP levels in prostatic tissue after androgen treatment in vivo.


HCG-dependent regulation of gonadotropin receptor sites: negative control in testicular Leydig cells. A single injection of human chorionic gonadotropin (hCG 500 IU) to prepubertal male rats increases plasma testosterone level and decreases hCG receptors in the testicular Leydig cells for more than 120 h. Injected hCG, measured in plasma using a specific radioreceptor assay for gonadotropins, is maximal at 2 h and decreases thereafter with an apparent half-life of 16 h. Plasma testosterone exhibits a rapid increase (30-40 ng/ml) within 1h after hCG injection. A delayed paradoxical increase (20-30 ng/ml) is observed between 48 and 120 h after the injection. The number of hCG binding sites in the isolated Leydig cells membranes decreases to less than 10% of the control value within 10 h and remains almost undetectable until 96 h after hCG injection. Reappearance of the binding sites is observed around 120 h. Similar, but less pronounced effects are found after the injection of 10 IU hCG. Since receptor occupancy cannot explain such a phenomenon, it is concluded that hCG is exerting a negative control on its own receptors in the Leydig cells.


Active transport of iodide in isolated porcine thyroid cells. Application to an in vitro bioassay of thyrotropin. Porcine thyroid cells were cultured for 2 days with or without dibutyryl cyclic AMP or thyrotropin (TSH). Then they were isolated post-culture by a gentle treatment with a calcium chelating agent. Some characteristics of the iodide transport system were studied in these thyroid cell suspensions. Iodide influx is a saturable, temperature- and energy-dependent phenomenon. It is blocked by ouabaïn, N-ethylmaleimide, dinitrophenol, cardiotoxin, low Na+ concentration and harmaline. Only 3% of the intracellular iodide is trichloracetic-acid-insoluble at equilibrium. The apparent Michaelis constant (Km) of the transport system is 30 micronM. For monolayer cells, the decrease of C/M ratio, increase of apparent Km, and decrease of Vmax between day 0 (freshly isolated cells) and day 6, indicate a loss of iodide-trapping ability up to passive diffusion. To the contrary, high values of C/M and normal Km (30 micronM) are observed in TSH follicles from reassociated cells. At iodide equilibrium, thryotropin, prostaglandins E1 and E2 and long-acting thyroid stimulator (LATS), induce a fast release of iodide. This release is dose-dependent in the first 5 min. It has been used to develop a bioassay of TSH and a fast detection of LATS.


Thyrotropin-specific binding to human peripheral blood monocytes and polymorphonuclear leukocytes. [125I]Labeled thyrotropin binding to leukocytes has been studied using lymphocyte-, monocyte- and polymorphonuclear leukocytes-enriched preparations obtained by centrifugation of Ficoll-Angiocontrix gradients and Sephadex G-10 adherence. From the relation between thyrotropin binding and phagocytosis as shown by latex beads ingestion, it is concluded that the hormone binds essentially to monocytes and polymorphonuclear leukocytes. Equilibrium association constants of the high-affinity, low-capacity sites (1 nM-1) are similar to those found in isolated thyroid cells or in thyroid plasma membranes. The role of thyrotropin in the regulation of phagocytosis by leukocytes is discussed.


Binding characteristics of L-triiodothyronine to isolated rat liver chromatin. The binding characteristics of [125I]-labeled L-triiodothyronine (T3) to chromatin isolated from rat liver nuclei were investigated. Binding of T3 to chromatin showed temperature-, incubation time-, and DNA concentration-dependence. According to Scatchard analysis, the apparent equilibrium dissociation constant was 225 pM, with a maximum binding capacity of about 0.2 pmoles per mg DNA. Displacement studies with unlabeled thyroxine (T4) and T3 showed that the binding sites for T4 might be the same as T3 but the binding affinity of the former was less than that of the latter. The binding was completely inhibited by the eukaryotic RNA polymerase inhibitor, rifampicin AF/021, but not by the prokaryotic inhibitor, rifampicin and alpha-amanitin. These observations indicate that the receptors for T3 have certain properties in common with RNA polymerase or other enzyme proteins which are sensitive to the rifampicin derivative. The hormone--chromatin fragments complex was solubilized from residual chromatin by digestion with DNase, but not with RNase, suggesting that the T3 receptors localize in the DNase-sensitive regions of DNA in the chromatin. This provides a useful method to use to investigate the localization of the receptor proteins in the chromatin and the interaction of the hormone-receptor complex with DNA.


Monozygotic triplets with discordance for diabetes mellitus and diabetic microangiopathy. A set of monozygotic triplets (PE.K., P.K., S.K.) has been studied. There is no diabetes in first-degree relatives. PE.K. developed insulin-requiring (60 U. NPH) diabetes at the age of 13 years. Over a period of 11 years since that time, numerous studies of insulin and growth-hormone secretion were performed on P.K. and S.K., including multiple oral glucose tolerance tests (OGTTs), cortisone-primed oral glucose tolerance tests (C-OGTTs), intravenous glucose tolerance tests (IVGTTs), and intravenous tolbutamide tests (IVTTs). The results of each test were compared with age- and sex-matched control subjects. P. K. developed insulin-requiring (56 U. NPH) diabetes after remaining discordant for eight years. Glucose, insilin, and growth-hormone responses during all tests were normal except during the IVGTT performed four months prior to the onset of diabetes. This last IVGTT revealed a glucose disappearance rate of 0.98 per cent per minute, and the slope of the regression line of serum-insulin response (IRI) on blood glucose (BG) was markedly decreased to 0.005 micronU./ml. IRI/mg./dl. BG (controls 0.340 +/- 0.04; mean +/- S.E.M.). The insulin responses in P.K. and S.K. were similar during all OGTTs, C-OGTTs, and IVTTs. S.K. has continued to maintain normal glucose tolerance and normal insulin and growth-hormone responses during all tests. The histocompability antigen studies have revealed HLA-A2, AW24, BW15, and BW40 phenotype in these monozygotic triplets. Muscle capillary basement membranes of the nondiabetic triplet were normal, whereas both diabetic triplets manifested evidence of capillary basement membrane thickening. The clinical and biochemical profiles in these triplets and the capillary basement membrane data lend strong credence to the role of "nongenetic" determinants in the development of "genetic" diabetes as well as diabetic microangiopathy in juvenile-onset-type diabetes.


Potentiation of the hepatic action of insulin by chlorpropamide. In perfused livers of fed rats, chlorpropamide inhibits glucagon-stimulated glucose production by augmenting the action of insulin. This effect is associated with a decrease in cyclic AMP accumulation in liver and perfusate. Alterations in glucose production appear to correlate more closely with changes in the amount of cyclic AMP in the perfusate than with changes in intrahepatic concentration of nucleotide. Potenitation by chlorpropamide of the hepatic action of insulin does not require administration of the drug prior to perfusion. Further, it is demonstrable at concentrations of insulin and glucagon (10(-11M) that approximate the normal plasma levels of these hormones.


Metabolic approaches to studies on diabetic microangiopathy. The chief purposes of this report are (a) to focus attention on various metabolic and pathophysiologic parameters relating prostaglandins (PGs) and thromboxanes to the slow but inexorable progression of vascular and blood cell dysfunction in diabetes mellitus and (b) to suggest areas of investigation that may be of fundamental importance for expanded areas of diabetes research. The prime thrust of these investigations would be to correlate these metabolic and pathophysiologic parameters with the vasculopathy of diabetes mellitus.


Sleep state, apnea and bradycardia in pre-term infants. Respiratory pattern and heart rate were studied in 28 pre-term infants and four full-term infants during the first weeks of life. Sleep state was identified by standard neurological and EEG criteria. Apneic spells lasting 10 seconds or more were most frequent in the least mature infants, and their frequency tended to diminish with increasing maturity. When these apneic attacks were related to sleep state they were found to occur more frequently during non-REM sleep in infants of 30 to 33 weeks gestation. This was the only statistically significant difference found. No over-all pattern relating sleep state to frequency of apnea could be detected for individual infants. It was also difficult to relate the occurrence of bradycardia to apnea in terms of sleep state or gestational age. It is concluded that no simple relationship between sleep state, apnea and bradycardia exists and that a complex interrelation between central and peripheral factors is involved in these events.


Effect of experimental diabetes and glucagon on cAMP-dependent protein kinase in rat liver. Liver protein kinase was determined in the absence and presence of cAMP4. Experimental alloxan diabetes resulted in a decrease in total protein kinase (+cAMP) and an increase in the activity ratio (-cAMP) divided by (+cAMP) in liver. Insulin treatment of diabetic rats reversed the observed changes in protein kinase in liver. Glucagon administered in vivo to normal rats caused an increase in the activity ratio and a decrease in total protein kinase activity in liver. The changes are similar to those in diabetes. A decrease in the ratio of insulin to glucagon in diabetes may account for the changes in protein kinase observed.


Permissive effect of glucose on the glucagon-induced accumulation of cAMP in isolated rat pancreatic islets. Glucose stimulation increased the cAMP content of collagenase-isolated rat pancreatic islets fourfold above baseline values. The elevation was transient, lasting about 5 min, and was dose-dependent. Insulin release continued at a constant rate throughout the incubation. Glucagon, in the absence of glucose, increased cAMP for about 1 min but only slightly, and had no effect on insulin release. In the presence of glucose, however, glucagon enhanced islet cAMP content 15-fold and increased the release of insulin. Glucagon was most effective at high glucose concentrations (16.6 and 25 mM). This indicates that glucagon is critically dependent on the presence of glucose in order to increase the islet cAMP content and to stimulate insulin release. The inability of glucagon to generate sufficient cAMP in the absence of glucose might be one of the reasons why the hormone is a potentiator rather than an initiator of insulin release.


A spontaneous transplantable osteogenic sarcoma in AKR/Ms mice. A transplantable osteogenic sarcoma originally arising in the right femur of an AKR/Ms male mouse is described. The original tumor showed a conspicuous bone and cartilage formation, but the capability of forming the bone was lost in the 4th transplant generation and that of cartilage formation was lost in the 7th generation. Alkaline phosphatase activity was histochemically demonstrated in only a few tumor cells, and the activity did not rise in the host serum. Intracisternal type-A particles of an average diameter of 70 nm were abundant in the rough endoplasmic reticulum, while type-C particles were rarely found to be budding from the cell surface or free in the extracellular spaces by electron microscopy. Three of the 27 thymectomized AKR mice that had been neonatally injected with cell-free material of the transplanted tumor developed osteomas, but no osteogenic sarcoma was found.


[Aspecific inhibitors of rubella virus hemagglutination in normal and thyroidectomized rats]. Thyroidectomized rats show alterations in serum lipoprotein distribution with an high increase of low density lipoproteins (LDL): however inhibitors titre was the same both in control and in treated animals. Also removal of nonspecific inhibitors by precipitation with heparin-MnCl2 or D.S. CaCl2 methods did not show differences between control and treated animals.


[Hemagglutinating activity of various viruses on trypsinized erythrocytes]. Mengo, Vesicular stomatitis and Semliki forest viruses titered with trypsin modified mouse, goose, pigeon and human erythrocytes show HA titers 2-32 fold higher than with the same untreated cells.


Antibody to hepatitis B core antigen in patients with hepatocellular carcinoma. Hepatitis B surface antigen (HBsAg), anti-HBs, and anti-HB core (HBc) were measured in 124 patients with hepatocellular carcinoma (HCC) in comparison with 299 control subjects of comparable ages, and in 48 cases of chronic hepatitis and 52 cases of hepatic cirrhosis. It was found that 72.6% of the HCC patients were positive for anti-HBc, and 80.6% were positive for at least one test, whereas in the control, anti-HBc was positive in 30.1% and 34.1% were positive for at least one test, the differences between the two groups being significant (P less than 0.01). The frequencies of positive tests for HBsAg and anti-HBc were the highest in HCC followed in decreasing order by cirrhosis, chronic hepatitis and the control group. A possible role of HB virus infection in hepatocellular carcinoma is discussed in relation to other factors.


Cytomegalovirus inclusions in patients with ulcerative colitis and toxic dilation requiring colonic resection. Microscopic examination of 50 colon resection specimens from 46 patients with ulcerative colitis revealed 6 patients whose colons contained intranuclear inclusions characteristic for cytomegalovirus (CMV). All patients were male, 47 years of age or older, and all had marked severe and fulminating courses. Five of the 6 patients had toxic dilation of the colon. Three of our patients had received no steroid therapy before their toxic dilation, however, all received steroids during attempts at medical management. Two other patients were found to have toxic dilation of the colon among our 46 patients, however, no evidence of CMV could be found in these 2 cases. There were 6 patients without toxic dilation whose colon specimens showed destructive inflammation and granulation tissue extending into the muscularis propria or transmurally. None of the colons from these 6 patients contained CMV inclusions. In our patients, the finding of CMV may be coincidental, but it is suggested that in our group of patients with ulcerative colitis, the CMV may play a role in altering the clinical course of these patients.


Transpyloric prolapse of polypoid gastric carcinoma. Four patients with discrete intraluminal filling defects in the duodenal bulb secondary to transpyloric prolapse of polypoid gastric carcinoma are reported. The lesions were pedunculated in 2 cases and sessile in the remaining 2. The clinical, radiological,and pathological findings are discussed. Prolapsed gastric carcinoma should be included in the differential diagnosis of localized intraluminal filling defects in the duodenal bulb and endoscopy and biopsy performed in patients with appropriate clinical and radiological findings.


The influence of bacillary dysentery on the efficiency of oral poliovaccine in Egypt. The rate of poliovirus excretion was found to be 64.6% in a group of 108 normal children and 11.95% in 184 diarrheal children. Diarrhea was due to Shigella in 70% of cases. This drew our attention to the presence of an etiologic relationship between the two findings, which may have a direct effect on the low efficiency of oral poliovaccine in our Country. Effect of Shigella infection on the take of oral poliovaccine was investigated in 14 normal and 10 children having acute diarrhea due to Shigella. Vaccine virus excretion was detected in 64.2% of normal children and only in 10% of diarrheal children. In vitro studies were done to determine the effect of killed Shigella suspensions or their endotoxin on the replication of poliovirus in tissue culture. Reduction of virus titre from 1-3 logs in the presence of killed Shigella or its endotoxin was observed. The sites of action and interfering factor (s) are discussed.


[The excretion of chorionic gonadetropin in 55 cases of hydatidiform mole (author's transl)]. In 55 cases of hydatidiform mole, 323 quantitative assays of chorionic gonadotropin were evaluated. In cases of molar degeneration the mean HCG excretion was 20% below normal. In cases of hydatidiform mole and invasive mole the mean HCG excretion was significantly above normal. Many cases of hydatidiform mole and all cases of invasive mole showed a repeat rise of the HCG titres after curettage. The pattern of HCG excretion in hydatidiform mole is shown in diagrams.


Age-related alterations in membrane lipid and protein interactions: Arrhenius studies of microsomal glucose-6-phosphatase. Arrhenius plots of glucose-6-phosphatase (EC 3.1.3.9) activity in liver microsomes from 6-month-old rats (young) showed discontinuities at 39, 30, 20 and 12 degrees C and change in activation energy at 20 degrees C. The enzyme activity in kidney microsomes of young rats showed essentially the same thermal discontinuities. In liver microsomes of 24-month-old rats (old) the enzyme showed discontinuities at 38, 30, 24 and 16 degrees C but not at 12 degrees C and the activation energy changed at 24 degrees C. In kidney microsomes of old rats discontinuities were seen at 38, 31, 22, 16 and 12 degrees C and the activation energy was constant from 0 to 41 degrees C. These results indicate that the interactions of membrane components are altered with age.


Transmission of rotavirus gastroenteritis from children to a monkey. A pooled suspension of rotavirus was prepared from the stools of eight children with acute non-bacterial gastroenteritis. The suspension was infused into the duodenum and stomach of an infant monkey (Nemestrina macaque). Biopsy samples of duodenal mucosa were taken at several intervals after inoculation, examined by light and electron microscopy, and assayed for lysosomal activity. Virus-like particles were seen within and around microvilli and intracellularly within vesicles as early as 20 minutes after the infusion. On the fourth and fifth days, large lysosomal bodies containing numerous virus-like particles were found within epithelial cells of the duodenal villi. No such particles were seen in the pre-inoculation sample or at days 16 or 25 after infection. The present study would appear to be the first demonstration of the transmission of this human virus to another species.


Certain aspects of the use of new method of microscopy in oncology. Electron microscopic studies on human neoplasms demonstrated that the typical cells fo individual neoplasms retain the ability of showing their specific ultrastructural differentiation. This fact provides theoretical basis for the use of electron microscopy in oncological practice for purposes of differential diagnosis. It also allows rejection of the concept of "dedifferentiation" and ultrastructural "simplification" as main ways of formation of neoplastic cells.


Ultrastructural localization of enzymes of the membranes and RNP-structures of the nuclei of normal and neoplastic cells. Ultrastructural localization of some oxidative and dephosphorylating enzymes was investigated in isolated rat and mouse liver nuclei and in nuclei isolated from various rat hepatomas. In the discussion the significance of changes in the activity of these enzymes in neoplastic cells is considered.


Hydrolysis of nucleoside triphosphate in plasma membranes of the hepatocytes of normal, regenerating and foetal livers and in cancer cells of hepatomas. Electron histochemical studies show that changes in the nucleoside triphosphatase activity in plasma membranes of cancer cells can proceed in different directions. Some cells show a high activity of magnesium-dependent NTPase over the whole membrane surface (perimeter), while others have a low enzymic activity which is present only in certain  regions of the membranes, the remaining cells possessing no enzyme activity at all. These changes are not strictly characteristic of cancer cells alone.


Histochemical methods in the diagnosis of precancerous states and early forms of cancer of the uterine cervix. Histochemical characteristics of displasia, various forms of the cancer in situ and initial invasive cancer together with histochemical criteria for differential diagnoses of these processes are given.


Ultrastructural localization of cytoplasmic phosphatases in preimplantation mouse embryos. The appearance and localization of the cytoplasmic phosphatases [acid phosphatase (AcPase) as a marker of lysosomes, TPPase as a marker of the Golgi apparatus, and NDPase (IDPase) as enzymatic marker of the endoplasmic reticulum (ER)] were cytochemically studied on the ultrastructural level in secondary oocytes and in preimplantation mouse embryos. The detectable AcPase activity, located on the inner surface of the membrane delimiting some cytoplasmic vacuoles (lysosomes and autophagic vacuoles), appears at the eight-cell stage and grows pregressively stronger up to the blastocyst stage. Golgi-associated reaction for TPPase was detectable in oocytes, dropped in one-cell embryos and became negative in the two-cell embryos. The reaction for TPPase and IDPase was present in plasma membranes of oocytes and early embryos and appeared in the delimiting membrane of some cytoplasmic vesicles in eight-cell embryos. Some activity of IDPase was found in small segments of the ER at the morula and blastocyst stage. The observed results suggest that the lysosomes are the first organelles in early embryos showing activity of the marker enzymes of the phosphatase type, while the activity of other marker enzymes is mainly concentrated in the plasma membrane of blastomeres. It cannot be excluded, however, that positive reaction for TPPase and IDPase in the plasma membrane results from nonspecific action of other phosphatases.


Activities of oxidative enzymes in thyroid follicles of Xiphophorin fishes. The activity of some intracellular oxidative enzymes was studied histochemically in the cells of the thyroid follicles of teleost fishes of the genus Xiphophorus. The experimental material consisted of animals of the red swordtail and Mexican swordtail breeds of Xiphophorus helleri and of melanotic Xiphophorus maculatus fishes. Observations were carried out on adult specimens of both sexes, including pregnant femals of Mexican swordtail. Moreover, immature Mexican swordtails of both sexes were examined. Thyroid follicles were found to be present in the subpharyngeal region of all fishes studied. The distribution of these follicles as well as their number and form depended on sex, age and on the analysed stage of prenancy. A smaller number and size of thyroid follicles were characteristic of immature specimens, whereas they were most numerous in the thyroids of pregnant fishes. The follicles were arranged in characteristic dense aggregations, especially in the melanotic platyfish. The follicular eipthelium in the fishes under study was usually cubical, but pregnant and non-pregnant adult females also contained a considerable number of larger follicles with flattened epithelium. Besides, thyroid follicles of multilayer epithelium were rather frequently encountered, especially in male fishes, irrespective of their age. The thyroid follicle cells of these fishes demonstrated invariably high activities of reduced NAD and NADP dehydrogenases and of beta-hydroxybutyrate dehydrogenase, and a low activity of succinat dehydrogenase. The intensities of alpha-glycerophosphate and lactate dehydrogenases and of cytochrome oxidase varied with sex, age and breed of the studied fishes. The immature and pregnant fishes showed the most clearly pronounced differences in the intensity of enzymic activity, the thyroid follicles of immature specimens revealing a high activity of lactate dehydrogenase and low activity of cytochrome oxidase, an inverse picture being seen in pregnant fishes. The adult forms of both sexes exhibited an enhanced activity of cytochrome oxidase and a decline in that of lactate dehydrogenase. The observed differences in the intensities of enzymic acitivities in the thyroids of the studied fishes are related with functions of this gland which in the period of growth are different from those in the period of sexual maturity, and certainly also with individual metabolic characteristics of the studied fishes.


The ammocoetes endostyle: its oxidative enzymes as an evidence of its homology with the thyroid of higher chordates. Histochemical methods were used for the demonstration of activity of the following intracellular oxidative enzymes, unstudied hitherto, in the epithelial cells of the endostyle of the river lampre (Lampertr aluviatilis L.) ammocoetes: reduced NAD dehydrogenase (NADD), lactate dehydrogenase (LD), cytochrome oxidase (CO), succinate dehydrogenase (SD), alpha-glycerophosphate dehydrogenase (alphaGPD) and glucose-6-phosphate dehydrogenase (G6PD). The activities of NADD and LD in the iodophil and throidogenic cells of type 3, then of subtype 2c and partly types 4 and 5 of the endostylar epithelium and the hypobranchial duct-lining epithelium were particularly ithe larva proves the possibility of their participation in the formation of the thyroid gland in the period of metamorphosis. In type 1 cells of the ammocoetes, despite their fairly strong enzymatic reactivity, the oxidative activity does not change significantly during the ontogenetic stages examined. The data obtained make it possible to modify the present views on the genesis of the thyroid gland of the adult  lamprey, namely, they indicate the participation of the type 6 cells of the hypobranchial duct-lining epithelium in the process of thyroidogenesis.