Natural history of valvular pulmonary stenosis in children below the age of two years. Long-term follow-up with serial heart catheterizations. 18 patients with mild and moderate valvular pulmonary stenosis diagnosed by heart catheterization before the age of 2 yr (mean 11.6 mth) were followed for 5 to 21 yr without operation. 15 cases had a repeat catheterization study after a mean interval of 10.7 yr. Of 7 cases with an initial right ventricular peak systolic pressure (RVSP) of 50 mm Hg or below, 6 had a decrease and 1 a moderate increase in RVSP. Of 8 cases with an initial RVSP of greater than 50 mm Hg 2 had a decrease, 2 had no change and 4 had a significant increase in RVSP; the latter subsequently underwent operation. Electrocardiography yielded some information, but neither roentgenography nor history was of any help in evaluating the severity of the stenosis at follow-up. As mild and moderate valvular pulmonary stenosis in infants and children below the age of 2 may progress and require operation a repeat heart catheterization is usually to be recommended later in childhood.


Expression of Thy-1 glycoprotein on lectin-resistant lymphoma cell lines. Lectin-resistant mutants with specific defects in glycosylation have been selected from the mouse lymphoma cell line, BW5147 (Thy-1+). The quantitative expression of cell surface glycoproteins on the mutant cells has been studied. The results show that some glycosylation defects that confer resistance to the cytotoxic effects of concanavalin A block the expression of Thy-1 glycoprotein on the cell surface. However, some changes in the oligosaccharides of Thy-1 glycoprotein generated by glycosylation defects found in PHAR mutant cells and restricted to the termini of complex-type oligosaccharides have no effect on the ability of Thy-1 to reach the cell surface. No glycosylation defects were found that interfered with the expression of either gp 69, 71 or H-2 on the surface of the mutant cells. It is concluded that aberrant biosynthesis of Thy-1 oligosaccharides can interfere with its expression on the cell surface, but that specific changes in oligosaccharide structure are necessary to block transport to the cell surface and integration into the plasma membrane.


Specific antigens of chicken thymus.

Specific antigens of chicken thymus. Purified plasma membranes from chicken thymus and bursa cells were prepared and solubilized with sarkosyl (sodium salt of N-methyl-N-(1-oxodecyl)-glycine). Antisera to solubilized thymus plasma membrane (TPM) were produced in rabbits and the globulin fraction obtained by ammonium sulfate precipitation. Four precipitating antigens were detected in solubilized TPM by immunoelectrophoresis. Following absorption with chicken serum and bursa plasma membrane (BPM) immunosorbents, three antigens, designated T1, T2, T3, were specific for the TPM fraction, and one antigen, T1, was found in soluble extracts of thymus tissue. Absorption with isolated plasma membrane and whole cells indicated that the T1 and T2 antigens in solubilized TPM are associated with the plasma membrane but not expressed on the surface of the cell. A common antigen, designated BT, was detected in BPM and TPM fractions and in membrane preparations of spleen. The antigens were not detected in any other tissues or cells including brain, circulating lymphocytes and erythrocytes.


The antiarrhythmic action of amitriptyline on arrhythmias associated with myocardial infarction in dogs. The antiarrhythmic activity of amitriptyline, a tricyclic antidepressant, was evaluated in anesthetized dogs 24 h after coronary occlusion, during the period of spontaneous ventricular arrhythmias. In all experiments amitriptyline was administered i.v. in incremental doses of 0.3 mg/kg at 1 min intervals until a conversion to normal sinus rhythm was evident. Amitriptyline administration resulted in conversion of the ventricular arrhythmia to a normal sinus rhythm in 100% of the animals tested at a mean dose of 1.3 +/- 0.1 mg/kg. Smaller doses also resulted in a dose-related decrease in non-sinus nodal pacemaker activity. Lidocaine, when administered to the same group of animals, produced a reduction of ectopic pacemaker activity, but did not eliminate it at a cumulative dose of 2 mg/kg. Antiarrhythmic doses of amitriptyline did not produce significant changes in arterial blood pressure, cardiac output or electrocardiographic parameters associated with atrioventricular or intraventricular conduction. The results of this study suggest that at very low doses amitriptyline may be an effective antiarrhythmic agent in ventricular arrhythmias associated with myocardial ischemia.


The effect of substantia negra stimulation and morphine on alpha-motoneurones and the tail-flick response. Rats were used to study the effect of unilateral stimulation of the substantia nigra on the reflex discharge of alpha-motoneurones and on  the reaction time of the tail-flick response. In preparations with prenigral decerebration, nigral stimulation facilitated monosynaptic alpha-reflex activity, whilst gamma-reflex activity remained unchanged. The facilitation of monosynaptic alpha-reflex activity was reduced by naloxone (1 mg/kg); morphine (2 mg/kg) did not change the number of alpha-reflex discharges, but it reduced the alpha-reflex latency, enhanced the effect of nigral stimulation on the latency and abolished the effect of naloxone on nigral facilitation. Nigral stimulation prolonged the reaction time of the tail-flick response in rats with an intact brain and after prenigral decerbration. Naloxone did not influence the anti-nociceptive effect of nigral stimulation, whilst morphine enhanced it in rats with an intact brain. The anti-nociceptive effect exerted by morphine in animals with an intact brain was abolished by prenigral decerbration, and an additional spinalization restored it. Inactivating the nigral neurones by unilateral microinjections of procaine or GABA into the substantia nigra depressed the nociceptive reflex. It is concluded that (1) activation of nigral neurones influenced mono- and polysynaptic reflexes in a reciprocal fashion by a pathway descending via brain stem relays to the spinal cord, (2) inactivation of nigral neurones produced similar changes in reflex activity by altering the function of the nigro-striatal feedback system, the outlet from the system to the spinal cord not being the substantia nigra, (3) morphine influenced the nociceptive reflex by an action at different levels of the central nervous system.


Aspirin- and indomethacin-induced ulcers and their antagonism by antihistamines. Gastric ulceration produced by aspirin and indomethacin was compared in acutely stressed and non-stressed rats. We found a synergism between these anti-inflammatory agents and acute stress in the production of gastric ulcers. Even at relatively high doses, neither agent caused appreciable gastric damage in non-stressed rats, whereas moderate doses of both agents produced massive ulceration in stressed rats. The synergism appears unrelated to the effect of these agents on the pituitary-adrenal response. The size and regional distribution of ulcers produced by aspirin and indomethacin in stressed rats were comparable. However, the dose--response curves of the two drugs were markedly dissimilar. Furthermore, the ulceration produced by indomethacin was attenuated by both H1 and H2 histamine receptor antagonists, whereas ulceration produced by aspirin was attenuated only by an H2 antagonist. The results suggest that the ulcerogenic mechanism of indomethacin may differ from that of aspirin and add to the growing evidence on the importance of endogenous histamine in various forms of gastric ulceration.


Application of high-powered microwave irradiation for acetylcholine analysis in mouse brain. Acetylcholine (ACh) levels in whole brains or discrete areas of mice brains were measured after sacrificing by decapitation, conventional or high-powered microwave irradiation. ACh levels in mice brains varied according to the time required to inactivate the enzymes. ACh levels after high-powered microwave irradiation for a shorter time were significantly higher than those after conventional irradiation.


Effect of labetalol on the accumulation and release of noradrenaline in rat ventricle. In rat ventricular tissue, labetalol inhibited the accumulation of (-)-[3H]noradrenaline and released [3H] following preloading with (-)[3H]noradrenaline. Cocaine (30 micrometer) inhibited the release observed with tyramine (5 micrometer) and beta-phenethylamine (5 micrometer) but not that observed with labetalol (5 micrometer). Reserpine pretreatment of the animals abolished the release observed with labetalol (5 and 50 micrometer). Labetalol primarily increased the loss of deaminated metabolites of noradrenaline. It is suggested that labetalol may release (-)-noradrenaline from the vesicles.


Chronotropic effect of tyramine on rat heart cells cultured with sympathetic neurons. Dissociated newborn rat superior cervical ganglion neurons in culture without exogenous nerve growth factor survive and extend processes on a monolayer of rat heart ventricular cells. An increase in the contraction rate of the heart cells was observed in 83% of the co-cultures treated with 5 X 10(-6) M tyramine. No increase was seen in heart cell cultures without neurons. These results are consistent with the assumed mode of action of tyramine--the release of catecholamines from nerve terminals--and suggest that functional interaction can occur in culture between sympathetic neurons and heart ventricular cells.


A new in vivo method for quantitatively evaluating local anesthetics. A method utilizing the reflex contraction of the retractor muscle of the hamster cheek pouch was developed to evaluate onset and duration of blockade produced by a local anesthetic agent infiltrated into or absorbed through the pouch epithelium. It provides a quantitative measure of reflex tension developed by the muscle as a function of afferent nerve conductivity in the pouch. The method is based upon the fact that when the pouch is everted activation of the retractor muscle results in the pouch being pulled toward the oral cavity. The method was validated by comparing the results with those obtained with other animal models and clinical trials. Ketocaine, an experimental drug, was evaluated for its efficacy as a local anesthetic when administered topically. In its present formulation the long latency of onset of blockade renders it impractical for clinical use.


Morphine: effects on serotonergic neurons and neurons in areas with a serotonergic input. The hypothesis that morphine acts on the serotonergic system to produce analgesia is based on the previous observations that (1) lesions and stimulation of midbrain raphe nuclei after the threshold to nociceptive stimuli; (2) morphine alters the turnover of serotonin (5-hydroxytryptamine; 5-HT). Microiontophoretic experiments were carried out to determine if morphine affected the firing rate of cells in five areas of the serotonergic system consisting of 5-HT containing neurons in the midbrain raphe nuclei (dorsal raphe and median raphe) or neurons in three areas (amygdala, optic tectum and subiculum) which are thought to receive a 5-HT input from the raphe nuclei. Morphine administered microiontophoretically slowed or inhibited the spontaneous neuronal firing in 34% of the cells studied in both the pre- and post-synaptic areas; systemically administered morphine gave similar results. However, the inhibition of neuronal firing by morphine in the five areas of the serotonergic system studied was not a specific narcotic effect because (1) it was not stereospecific; and (2) it was rarely blocked by naloxone. The inhibitory effect of morphine does not appear to be related to the analgesic effects of morphine because neuronal firing in the midbrain raphe nuclei and three possible postsynaptic areas was rarely altered by a nociceptive stimulus. It is concluded that (1) the analgesic effect of morphine is not related to an effect on neurons in the midbrain raphe nuclei and three areas which receive an identified 5-HT input; and (2) the effect of morphine on neurons in these five areas of the serotonergic system is not a specific narcotic effect.


Consummatory behavior and urine production after cerebroventricular injection of vasopressin and vasopressin antiserum. Vasopressin, or vasopressin antiserum, was injected into a lateral cerebral ventricle of conscious rats. These rats were normally hydrated, cellular dehydrated (NaCl loading) or hypovolemic (polyethylene glycol model). Elevation or reduction of vasopressin in cerebrospinal fluid produced no consistent change in consummatory behavior, urine volume or sodium and potassium excretion. These results show vasopressin in cerebrospinal fluid not to be an absolute requirement for maintenance of hydration or for response to acute volume and osmotic stimuli.


Effect of hypothalamic stimulation in spontaneously hypertensive and Wistar-Kyoto rats. This study was undertaken to determine if central nervous system differences in blood pressure regulation exist between spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) controls. Central control mechanisms were examined by observing the effects of posterior hypothalamic (PH) stimulation upon preganglionic sympathetic activity in 14--18 week old SHR and WKY rats. A bipolar, concentric electrode was stereotaxically placed in the PH. Stimulation was delivered at 20, 60 and 100 Hz (3-sec duration, 0.1 msec pulse width) at a voltage twice that producing an increase in blood pressure (less than 5 mm Hg) at 60 Hz. Sympathetic activity was recorded from a portion of the splanchnic nerve just distal to the diaphragm. Blood pressure was measured from a femoral artery catheter. SHR responded with greater increases in sympathetic activity than WKY; the differences were statistically significant at 60 and 100 Hz. SHR also responded with significantly greater increases in blood pressure at all frequencies of stimulation. To determine if the enhanced sympathetic response to PH stimulation seen in adult SHR is an intrinsic difference rather than secondary to sustained hypertension, we maintained SHR normotensive from four weeks of age with antihypertensive drug therapy (clonidine or hydralazine). Chronically treated animals were then tested at 14--18 weeks of age while on antihypertensives or four days after drug discontinuance. Sympathetic and blood pressure responses to PH stimulation were significantly greater in SHR maintained normotensive than untreated or chronically treated WKY. These data support the concept that a central factor is involved in the etiology of hypertension in the SHR.


Nucleus accumbens lesions reduce amphetamine hyperthermia but not hyperactivity. Electrolytic lesions of the nucleus accumbens were found to produce an increase in spontaneous locomotion; additionally, these lesions attenuated the hyperthermic, but not the hyperkinetic, actions of d-amphetamine. These results suggest that while dopamine release from the mesolimbic system may underlie amphetamine-induced hyperthermia, other structures must also be involved in amphetamine-induced hypermotility.


[Establishment of a new breeding colony of germfree CF no. 1 mice (author's transl)]. Baby CF no. 1 mice were obtained by hysterectomy and reared by hand feeding under aseptic condition. Temperature in the cage was kept at 33 degrees C for first 14 days of age. Artificial milk, diet RM was composed of rat milk (44%) collected from the stomach of killed baby rats, evaporated cow milk (16%) and vitamin mixture (2%). The diet was homogenized after sterilization and given mice using a latex gum stomach tube at intervals of four hours for 20 to 24 days of age. Feeding size a day (Y ml) was given by the following regression curve, Y = 0.412 X - 0.299, (X : body weights in gram). According to this procedure, 40 out of 74 germfree mice (54%) were weaned with good growth. From a male and a female mice, the germfree CF no. 1 mouse colony of Takeda was established in 1967 and the lines were distributed to several laboratories of this country.


[Measurement of rat serum corticosterone using radiostereoassay kit for cortisol (author's transl)]. A very simple technique for measurement of rat serum corticosterone with radiostereoassy kit using 75Se-cortisol was devised. Only 25--50 microliter serum was required for this assay. The serum sample was first diluted to 300 microliter with water and heated in a glass tube to denature the endogeneous cortisol binding protein. Then an aliquot of the sample was transferred into an assay vial containing 75Se-cortisol, cortisol binding protein and adsorbent granules. After rotated, an aliquot of the supernatant was taken for counting. High level of rat serum corticosterone was measured with satisfying accuracy by dilution with water. Serum corticosterone levels in 7 normal female Sprague-Dawley rats (7 weeks old, Ca 200g body weight) at 2.00 pm were 29.5 +/- 12.2 microgram/dl (mean +/- SD). The levels were lower and higher in rats treated with dexamethasone and with 1-18 ACTH, respectively.


[Survey of marker genes in the 9 rat strains (author's transl)]. In total of 17 genetic traits of the 9 strains of rats, Rattus norvegicus, examined, 9 loci showed polymorphic, being a, c, d, h, Es-1, Es-2, Es-3, Amy-1 and sex-influenced esterase (d-allele is not identified [12]. These genetic markers are not only useful as linkage markers but also available for inspection of proper maintenance of inbred rat strains.


[A simplified method for determination of biochemical marker genes in inbred strains of mice (author's transl)]. A rapid method is described for the determination of biochemical variants of mice by Titan III cellulose acetate electrophoresis. Variants examined were hemoglobin beta-chain, serum esterase-1, malic enzyme (supernatant form), and isocitrate dehydrogenase (supernatant form). The present procedure can be recommended as a simplified method to analyze biochemical variants for checking of marker genes in inbred strains of mice.


[Small scale efficient reproduction of El mice (author's transl)]. Epilepic anormaly strain of mice "El" derived from National Institute of Health, Japan were bred by sister-brother mating during the first 3 months and maintained thereafter by random mating at this laboratory. About 20% (21/101) of virgin females and 4% (2/49) of males were infertile and rate of delivering was relatively poor; mean delivery rate was 70.6%. Delivery rate were drawn after fourth litter. The mean litter size and weaning rate were 7.54 and 97.9%, respectively. Male El mice died following convulsion. Autopsy revealed the obstruction of urether, bleeding in the bladder and seminal vesicules. The time of 50% death was 166 days of age.


[A micro-method developed for prothrombin time assay (author's transl)]. Prothrombin time was measured by a newly developed micro-method using a plastic film available in the market (PARAFILM, American Can. Co.). The comparative study of this micro-method with that of Quick in rats disclosed a good correlation, with correlation coefficient of 0.951, supporting the usefulness of the method for examination of blood coagulability. The new method gave the physiological values of 9.8 sec in rabbits, 12.5 sec in dogs, 13.3 sec in mice, 14.8 sec in cats and 16.0 sec in rats, respectively. Among them, guinea pigs took the longest time of 25.3 sec for the coagulation.


Effect of a single oral dose of oxymetholone on the metabolism of human erythrocytes. Androgenic steroids have been shown to enhance erythrocyte 2,3-DPG production in vivo and in vitro, and to stimulate the pentose shunt oxidative reactions in vitro. Furthermore, a 3 beta- and a 17 beta-hydroxysteroid dehydrogenase have been identified in red cells. The present study was carried out to explore a cumulative effect of androgens on glycolysis and androgen reduction in human erythrocytes in vivo following a single 50 mg oral dose of 17 beta-hydroxy-2 (hydroxymethylene)-17 methyl-5 alpha-androstan-3-one (oxymetholone). The rate of erythrocyte glycolysis was measured by quantitative determination of: fructose-1,6-diphosphate (FDP); dehydroxyacetone phosphate (DAP); 2,3-diphosphoglycerate (2,3-DPG); adenosine triphosphate (ATP); and lactate. Serum and erythrocyte steroids were separated by thin layer chromatography. The reduction of 5 alpha-androstan-17 beta-ol-3-one by red cell hemolysate was measured in the presence of NADPH as an index of 3(17)beta-hydroxysteroid dehydrogenase activity. Our results  show that oxymetholone administration is followed by the appearance of an unidentified steroid fraction in chromatograms of serum and erythrocytes, simultaneously with the enhancement of glycolysis and of hydroxysteroid dehydrogenase activity in erythrocytes. A direct effect of androgen on erythrocyte metabolism, which is independent of the hormone erythropoietic effect, is discussed.


Population sizes of granulocyte-macrophage and monocyte-macrophage colony-forming cells in Sl/Sld. The sizes of granulocyte-macrophage (CFU-c) and monocyte-macrophage (CFC) progenitor cell populations were measured and compared in Sl/Sld and +/+ mice. In the marrow, the CFU-c and CFC population sizes were 40% and 67%, respectively, of the +/+ marrow CFU-c and CFC population sizes. There was no difference in the size of these two-cell populations in the spleens or thymi of Sl/Sld mice as compared with +/+ mice. Furthermore, the marrow CFC population was found to be heterogeneous by velocity sedimentation. One of the marrow CFC subpopulations is characterized by a velocity sedimentation value of 5.05 mm/h while the other has a value of 6.30 mm/h.


Effect of bacterial lipopolysaccharide on proliferation of CFU-S. Multiple injections of S. typhosa LPS increased the number of CFU-S in the spleen 20-50 fold and decreased the number in the femur to one half or less. LPS injections did not affect the growth rates of CFU-S in the spleen or femur of lethally irradiated mice. The plateau levels which were attained in these mice after proliferation, corresponded with the levels in LPS-treated non-irradiated mice. Local irradiation of the spleen with 3000 rad hardly affected the capacity of the spleen to accommodate the increased CFU-S numbers after LPS injection. These results suggest that irradiation resistant microenvironmental factors in the spleen determine the CFU-S accumulation in this organ after injection of Salmonella typhosa LPS. The increased number of CFU-S in the blood after LPS injection was maintained in splenectomized mice as well as in mice which received a local splenic x-irradiation with 3000 rad, indicating that this rise of blood CFU-S numbers is independent of the spleen.


Hemopoiesis on macrophage-coated cellulose acetate membranes (CAMS) in mice: an immunological study. Macrophage-coated cellulose acetate membranes (CAMS), implanted into the peritoneal cavities of sublethally irradiated mice, support the growth of hemopoietic colonies. To investigate the nature of the precursor cells (CFU-ML) which form colonies on CAMS, we pre-treated marrow cells with rabbit anti-mouse brain serum (RAMBS), a known anti-pluripotent stem cell (CFU-S) serum, plus complement (C) and studied the number of colonies formed and the distribution of their sizes among the various histological types. Marrow cells pre-treated with RAMBS + C, even with the opportunity for interaction with macrophages in vivo, did not form fewer or smaller colonies than those formed from CRS + C treated cells, suggesting that most of the CFU-ML are antigenically distinct from CFU-S.


Erythroid colony formation in vitro from the marrow of dogs with cyclic hematopoiesis: interrelationship of progenitor cells. Committed erythroid progenitor cells (Colony Forming Units-Erythroid, CFU-E) have been studied in canine cyclic hematopoiesis (CH) utilizing a semi-solid methyl cellulose culture system. Erythroid colonies were stimulated by the addition of a standard volume of serum from normal dogs that had been phlebotomized and subjected to hypoxia. CFU-E fluctuated over the cycle in dogs with CH from concentrations 4--5 times normal during the periods of peripheral blood neutropenia to less than one tenth of normal during the phases of elevated peripheral blood neutrophil counts. In spite of these marked fluctuations there was no change in the proliferation rate of the CFU-E as estimated by the tritiated thymidine (3H-TdR) suicide technique. Failure to demonstrate a change in the CFU-E proliferation rate may be related to the relative maturity of these cells with the fluctuations in number resulting from a 'feed-in' from more immature cells. The results show that CFU-E fluctuate in the same phase as committed granulocytic progenitor cells (CFU-C). Our current knowledge of the interrelationships of marrow progenitor cells and events in the peripheral blood of dogs with CH is briefly reviewed and some additional questions, raised by recent studies regarding the pathogenesis of this disease, are discussed.


Determination of heme and non-heme iron content of mouse erythropoietic organs. A procedure is described for determining the content of heme and non-heme iron in organs and tissues of the mouse. Heme iron after homogenization and hemolysis of the samples is extracted as hemin by means of a mixture of ethyl acetate and glacial acetic acid, the extraction being followed by washing with hydrochloric acid. Total iron content is determined with commercial analytical sets exploiting color reaction with bathophenanthroline after mineralization of the samples. The presence of bone or other tissues does not distort the results. The procedure is relatively simple and suitable for serial analyses. The results obtained for the mouse by this method are in good agreement with recognized principles of ferrokinetics in mammals.


Beta-adrenergic thirst and its relation to the renin-angiotensin system. Isoproterenol is a potent dipsogen and antidiuretic agent. It also stimulates the release of renin from the kidney. Evidence is presented to substantiate the view that the drinking and increased vasopressin release that follow the systemic injection of a small dose of isoproterenol are mediated via increased activity of the renin-angiotensin system. Larger doses of isoproterenol, which have profound effects on the cardiovascular system, cause drinking and vasopressin release by mechanisms that do not depend solely on the renin-angiotensin system. Other experiments discussed do not support the hypothesis that hypothalamic beta-adrenergic neurons are important in facilitating thirst. Low doses of isoproterenol are more effective in causing drinking and vasopressin release when given peripherally rather than centrally. Evidence is discussed that supports the view that isoproterenol given centrally leaks into the periphery and causes release of renin and subsequent stimulation of drinking and vasopressin release.


The renin-angiotensin system and thirst: some unanswered questions. Recent experiments dispute the significance of four findings that usually are cited to support the hypothesis that angiotensin is a natural dipsogen. First, although administration of exogenous renin consistently increases water intake, the plasma renin activities that are produced seem to be outside of the normal physiological range when the elicited drinking is substantial. Second, although plasma renin activities are elevated following caval ligation, colloid, or isoproterenol treatment, this activity of the renin-angiotensin system appears to account for only a small portion of the observed water intake. Third, although bilateral nephrectomy abolishes the water intake that otherwise occurs after caval ligation or isoproterenol treatment, the observed blood pressure appears to be so low as to preclude drinking behavior. Finally, although drinking can be stimulated in these hypotensive rats by various treatments, in each case a pressor response also is observed that might have restored the ability to drink. These new observations have provoked a reevaluation of the circumstances in which endogenous angiotensin stimulates thirst and the magnitude of its contribution then. At present, there is no persuasive evidence that the renin-angiotensin system normally plays a direct and substantial role in mediating the ingestion of water or saline.


Intestinal ischemia and shock factors.

Intestinal ischemia and shock factors. Perfusion pressure reduction to the small intestinal vascular bed for 2 hours to followed by characteristic villous damage and a general cardiovascular derangement. In an extensive series of experiments on cats it was demonstrated that the cardiovascular derangement could not be ascribed to any pooling of blood and/or fluid in the small intestine. Cardiotoxic material has been demonstrated in the cat intestinal venous blood in in vivo and in vitro experiments using working rat hearts and isolated rabbit papillary muscles. A relationship between the extent of the villous damage and the appearance of cardiotoxic material in the intestinal venous blood has been found. The cardiotoxic material seems to consist of at least two heat stable fractions. One is water soluble with a molecular mass between 500 and 1,000 daltons and the other is lipid soluble with an unknown molecular mass.


Human leukocyte inhibitory factor (LIF): a lymphocyte mediator with esteratic properties. Sensitized human or guinea pig lymphocytes stimulated by specific antigen or nonspecifically by mitogens elaborate a soluble leukocyte inhibitory factor (LIF) that selectively inhibits the random migration of polymorphonuclear (PMN) leukocytes. LIF has the properties of a protein and is unique among lymphocyte mediators because of its esterase nature. While its natural substrate on PMN leukocytes is unknown, a synthetic amino acid derivative ([3H]benzoyl arginine ethyl ester) has been shown to be capable of serving as an artificial substrate. Through the use of selective esterase inhibitors it may eventually be possible to develop a biochemical assay for its detection in lieu of the biologic assay. LIF-containing supernatants have been shown to enhance a number of PMN leukocyte functions, including adherence, phagocytosis, metabolism, and electrophoretic mobility. In a chemotactic assay, LIF was found to induce the release of a low molecular weight peptide (neutrophil immobilization factor) from PMN leukocytes which decreased their spontaneous random movement and directed movement in the presence of a chemoattractant. This latter pathway may provide an amplification of LIF activity on the migration of PMN leukocytes.


Failures following fimbriectomy: a further report. In 1977 we reported on seven patients who presented with intrauterine pregnancy in spite of previous fimbriectomy; in a detailed morphologic study one tuboperitoneal fistula was documented in each of four patients. In the present study three more patients are reported with pregnancy following fimbriectomy, and another four tuboperitoneal fistulas are morphologically investigated and their possible etiology is discussed. In spite of a failure rate of almost 2.6%, the concept about the reliability of fimbriectomy need not necessarily be condemned. This concept can be judged only if information is available both about the total number of fistulas subsequent to a certain technique and about the number of those fistulas which contributed to pregnancy.


Continued clinical experience with an increasing dosage regimen of clomiphene citrate administration. Between August 1, 1975, and July 31, 1977, 60 patients with infertility and ovulatory dysfunction were evaluated at the Naval Regional Medical Center, Oakland. Of these, 15 had incomplete work-ups or were lost to follow-up. Of the remaining 45 patients, 2 (4.5%) failed to ovulate at a maximal dose of 250 mg of clomiphene for 5 days with 10,000 units of human chorionic gonadotropin. Of the 43 (95.5%) who did ovulate, 33 conceived. Of these 33 patients, 9 (27%) required greater than 100 mg of clomiphene and/or longer than three ovulatory cycles to conceive. Of the 10 patients who did not become pregnant, additional infertility factors were present in 5. When these 5 were eliminated, the corrected pregnancy rate was 87% (33 of 38). The spontaneous abortion rate was 12.2% and there were no multiple gestations. These results would further emphasize that clomiphene can be used safely and effectively at doses greater than 100 mg or for longer than three ovulatory cycles.


Diagnostic evaluation of hirsutism in women by selective bilateral adrenal and ovarian venous catheterization. To determine the most significant secretory source of their androgens, 13 hirsute nonvirilized women underwent selective bilateral adrenal and ovarian venous catheterization to obtain effluent blood for the assay of testosterone and delta4-androstenedione. In three patients the testosterone and delta4-androstenedione gradients were significantly greater in the adrenal venous effluents. Testosterone and delta4-androstenedione gradients were significantly greater in the ovarian venous effluents in four patients. In six patients there were no significant differences in the testosterone gradients between the adrenal and ovarian venous effluents. The delta4-androstenedione gradients were greater in the adrenal venous effluents in three of these patients, greater in the ovarian venous effluents in one, and not significantly different in two of these patients. The fact that the measurement of urinary 17-ketosteroid excretion, the suppressibility of peripheral plasma androgens with dexamethasone, and the stimulation of peripheral plasma androgens with human chorionic gonadotropin correlated poorly with the selective catheterization data suggests that the former modalities are imprecise in the diagnostic evaluation of hirsutism in women.


The effects of a once-a-week steroid contraceptive (R2323) on lipid and carbohydrate metabolism in women during three months of use. Forty-four women were prospectively evaluated with a 3-hour oral glucose tolerance test while they used a new weekly oral contraceptive drug, R2323. The blood tests were carried out before the drug was administered and again 3 months after its use. Both blood glucose and plasma insulin levels were measured. In the 40 women with a "normal" control glucose tolerance test, it was found that only the 1-hour blood glucose value was significantly elevated after 3 months of drug use and the plasma insulin values were unchanged. The tests of three of the four women with borderline abnormal to abnormal control test curves converted to normal with 3 months of treatment and that of one remained unchanged. The fasting plasma cholesterol and triglyceride levels were unchanged over the 3 months' time. These results suggest that this steroid contraceptive has no effect on lipid metabolism. It also has minimal adverse effects on normal carbohydrate metabolism and may possibly improve abnormal metabolism.


Quantitation of Leydig cells in testicular biopsies of oligospermic men with varicocele. Leydig cell density was evaluated quantitatively in bilateral testicular biopsies from 16 oligospermic men with varicocele. The method utilized for this quantitation is based on the determination of the total number of Leydig cells, Leydig cell clusters, and seminiferous tubules in the entire histologic section of each biopsy, and the calculation of the following indices: mean Leydig cells per seminiferous tubule, mean Leydig cell clusters per seminiferous tubule, and mean Leydig cells per cluster. A significant correlation between Leydig cells per tubule and Leydig cell clusters per tubule was demonstrated, suggesting that determination of Leydig cell clusters per seminiferous tubule in testicular biopsies is an objective and clinically applicable method for quantitative evaluation of Leydig cell density. An association of varicocele with Leydig cell hyperplasia was not noted for the 16 patients studied. In addition, no significant differences in Leydig cell density between right and left testes were found, and no correlation between Leydig cell density and site or degree of varicocele, or age of the patients could be demonstrated. To our knowledge this is the first report of quantitative analysis of Leydig cell density in men with varicocele.


A new chamber for rapid sperm count and motility estimation. A new chamber for sperm count and motility estimation is described. This chamber, which is only 10 micron deep, enables free horizontal movement of spermatozoa in one focal plane and provides conditions for the examination of undiluted samples. Therefore, with the aid of this instrument it is possible to compare sperm motility in various samples from the same person or in different samples at different times. This can be done either by simple estimation or with any other method of motility evaluation chosen by the examiner. The sperm count can be made rapidly and directly from an undiluted, preheated sample by counting spermatozoa in the area of a grid located within the eyepiece; the count is expressed in millions per milliliter. Thirty-seven specimens were analyzed with this chamber. Statistical evaluation of the results revealed high precision, accuracy, and reliability of sperm counts when compared with the hemocytometric method. Better results were obtained when motility estimation was compared with the ordinary slide technique. Easy performance, rapid sperm counts, and improvement of motility estimation make this chamber a useful tool where sperm analysis is carried out.


Evaluation of a new silastic seminal fluid collection device. A new Silastic seminal fluid collection device has been tested and compared in vitro with glass jars, latex condoms, and the Milex polyethylene sheath. In a series of tests utilizing 50 different semen samples and observing the effects on motility over a period of 4 hours, the Silastic seminal fluid collection device demonstrated sperm survival slightly better than that with glass jars, superior to that with the polyethylene Milex sheath, and far greater survival (by several hours) than with the latex condom. The Silastic collection device was found to be more comfortable and more pleasurable to the patients than was the Milex sheath. The accuracy of seminal volume measurements was also found to be better in the Silastic collection device.


Binding of steroids to human spermatozoa and its possible role in contraception. The binding of steroids to human ejaculated spermatozoa and the effect of steroids bound to spermatozoa on sperm migration and motility in vitro was examined. A correlation between progestogens that bind to steroid-binding sites on human spermatozoa and progestogens that inhibit sperm migration was established. The results indicated that there is a direct and specific steroid effect on human spermatozoa, as some steroids such as progesterone, lynestrenol, and norethynodrel markedly inhibited sperm migration and motility, whereas other steroids such as estrone had no detectable effect on sperm migration and motility. The significance of these findings was discussed in relation to the contraceptive action of steroids applied directly to the lumen of the female genital tract.


The role of phosphate esters in male fertility. Spermatozoa do not achieve full maturation and fertilizing capacity until passage through the epididymis. During this time they also gain motility, although spermatozoa do not move until after ejaculation. The organic fraction of human seminal plasma contains phosphate esters, particularly glycerylphosphorylcholine (GPC), phosphorylcholine (PCh), and inorganic phosphate (Pi). GPC is found in relatively high concentrations in the semen of many male animals, including man. GPC is synthesized by the epithelial cells of the epididymis, apparently under androgenic control. Consequently, it has been suggested that GPC might be a useful indicator of epididymal function. We have measured GPC, Pi, and PCh in fresh and frozen semen samples, using phosphorus nuclear magnetic resonance (31P NMR). All samples were assayed for phosphate esters. It was found that PCh was totally hydrolized to Pi. The average ratio of GPC to total phosphate (TP = GPC + Pi) remained constant at a value of about 0.1 for sperm counts over 20 million/ml. The ratio for azoospermic specimens was 0.02 or less; the same results were obtained from vasectomy specimens. This finding indicates that most of the GPC comes from the epididymis. There was a significant correlation between motility, progression, and the GPC ratio. Poor motility and progression in the specimens were accompanied by low GPC ratios regardless of the sperm counts.


The effect of prostaglandins F2alpha and E2 on pregnancy in mice during implantation. The uterine content of prostaglandins (PGs) increases following insertion of an intrauterine device (IUD). Such IUD-bearing uteri also exhibit impaired decidualization. Although prostaglandins have been shown to be luteolytic in several species, it is not clear that inhibition of decidualization is mediated solely through alterations in ovarian steroidogenesis. In these experiments, ovariectomized, hormone-replaced pregnant mice, as well as intact mice, were treated systemically with either PGE2 or PGF2alpha so that the extraovarian actions of prostaglandins upon decidualization could be segregated from effects mediated by alterations in ovarian steroidogenesis. The results suggest that the effect of PGF2alpha upon decidualization was mediated via an alteration in ovarian steroidogenesis, but that PGE2 acted independently of the ovary. The hypothesis is presented that alterations in the ratio of E to F prostaglandins elicited within the endometrium by an IUD inhibit decidualization at the uterine level, and that this effect might be mediated by impairment of histamine release from endometrial mast cells.


The effect of intra-amniotic prostaglandin F2alpha on anterior pituitary hormone release during midtrimester abortion. The effect of intra-amniotic administration of prostaglandin F2alpha (PGF2alpha) on pituitary hormone release was studied in women undergoing midtrimester abortion. Serum prolactin (PRL), growth hormone (GH), thyrotropin (TSH), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were measured prior to and at 15-minute intervals for 2 hours following intra-amniotic administration of 40 gm of urea alone, 20 mg of PGF2alpha and 40 gm of urea, or an equal volume of normal saline. Serum PRL levels were significantly higher at sampling times from 15 through 120 minutes when both PGF2alpha and urea were administered than after saline or urea alone. The elevation in serum GH following PGF2alpha and urea was not significantly greater than for urea alone. The GH response in the women receiving normal saline was significantly less than for the two groups of women receiving the abortifacients. These results indicate that the GH response was related to the stress effects of the abortifacients. There was no difference in the TSH, LH, and FSH responses for the three groups. These results suggest that PGF2alpha selectively causes pituitary release of PRL in women during midtrimester pregnancy.


Effect of 15(S)-15-methyl prostaglandin F2alpha on human oviductal motility and ovum transport. The effects of an intravenous infusion of 15(S)-15-methyl prostaglandin F2alpha (PGF2alpha) on oviductal motility and ovum transport were studied in women who were scheduled for elective tubal sterilization. Infusion rates of 0.38 microgram/kg/hour or higher caused an increase in oviductal motility in all patients. Lower infusion rates did not always cause a stimulation of motility. Low infusion rates generally caused an increase in the amplitude of contractions without any effect on basal oviductal tone. The higher infusion rates usually caused a large increase in basal tone as well as an increase in the amplitude of contractions. Ova were recovered from the oviducts of five patients who had received an intravenous infusion of 15(S)-15methyl PGF2alpha. The ova were recovered from the ampulla in three patients, from the ampullary-isthmic junction in one patient, and from the isthmus in one patient. Since one would expect to recover ova from the oviducts at similar times under normal circumstances, there was no evidence that this prostaglandin treatment caused an acceleration of ovum transport. These data support the conclusion that a PGF analog which stimulates oviductal motility does not necessarily also accelerate ovum transport in women.


The duration of infertility: an important datum for the fertility prognosis of men with semen abnormalities. In a group of 584 men with semen abnormalities the relationship between conception and the duration of infertility was studied. "Spontaneous" pregnancies were reported significantly more frequently for men with an infertility duration of less than 2 years than for men with an infertility duration of more than 2 years. Seventy-three conceptions occurred "spontaneously" and sixty-six occurred while the men were undergoing different forms of therapy. When all pregnancies were considered together it appeared that the "chance" of fathering a child after 2 years of infertility was still about 20% during the ensuing 3 years.


Immunologic studies of male infertility. Blood and semen samples were obtained from 13 men who had self-agglutination of spermatozoa and spermatozoal antibodies in their sera. The seminal plasma of these men produced spermatozoal immobilization and agglutination when added to normal semen from healthy men. Analysis of the seminal plasma samples showed that immunoglobulin G (IgG) levels were higher in the study group than in the control subjects. They contained IgA and complements C3 and C4, whereas only 2 of 15 samples from normal men contained IgA, and only one case contained C3 and C4. IgM was detected in 10 samples of the study group and in one sample of the control group. IgG levels in the blood of the study group were significantly higher than those in the blood of control subjects. Therefore, the immunoglobulins which probably play a role in the mechanism of autoagglutination and immobilization of spermatozoa are IgG, IgA, and IgM, and complements C3 and C4.


Statisical comparison of the parameters of semen analysis of whole semen versus the fractions of the split ejaculate. The split ejaculate has proven to be a reliable method for concentrating sperm in a small volume. However, there is some controversy whether the spermatozoa in the most sperm-rich fraction of the split ejaculate has better motility than the spermatozoa in whole semen. In this study, at least two specimens of whole semen and two split ejaculates were obtained from 45 infertile males. The mean values for the parameters of the semen analysis were compared statistically. Our data indicate that increased sperm density is a consistent finding with the split ejaculate, even for severely oligospermic men. However, improved sperm motility with the split ejaculate is more variable. On the other hand, patients with mild oligospermia (10 to 40 million sperm/ml of whole semen) and high semen volumes (greater than 5.0 ml) demonstrated improvement in all parameters in the split ejaculate.


Suppressive effect of 1,2-dibromo-3-chloropropane on human spermatogenesis. Azoospermia was diagnosed in six factory workers who had been chronically exposed to 1,2-dibromo-3-chloropropane. Infertility was the presenting symptom in two patients and a decrease in libido or impotence characterized the others. Hormone studies revealed elevated plasma follicle-stimulating hormone levels and normal plasma luteinizing hormone and testosterone concentrations. Testicular biopsy showed selective atrophy of the germinal epithelium, intact Sertoli cells, and a normal appearance of a relatively increased number of Leydig cells.


Quantitative analysis of the seminiferous epithelium in human testicular biopsies, and the relation of spermatogenesis to sperm density. Quantitative analysis of the seminiferous epithelium was performed in bilateral testicular biopsy specimens from 14 patients with sperm counts ranging from 0 to 89 million/ml. All Sertoli cells and germ cells within each seminiferous tubule cross-section were counted in all biopsies. Results were expressed either as number of cells per unit length of seminiferous tubule circumference or as number of cells per Sertoli cell. Results were correlated with sperm count (millions per milliliter), total sperm count (millions per ejaculate), and age. A significant correlation between sperm density and germ cell counts was demonstrated. Coefficients of correlation were higher when results were expressed per unit of tubular wall length than when expressed per Sertoli cell. In men with sperm counts below 5 million/ml the number of germ cells in the biopsy was lower than in men with higher sperm counts. Spermiogenesis appeared to be most affected. In this group of patients an adverse effect of age on spermatogenesis was noted.


Short-loop feedback of luteinizing hormone: dose-response relationships and specificity. The short-loop feedback control of rabbit luteinizing hormone (rLH) was studied by using a highly specific radioimmunoassay (RIA) system for rLH which does not react with human LH. Permanent intravenous catheters were placed in adult female New Zealand White rabbits at the time of castration. Highly purified human luteinizing hormone (hLH) was injected intravenously at doses of 0.1, 0.3, 0.5, 1, 2, 3, 4, 10, 50, and 100 IU into unanesthetized animals 1 to 16 days following castration. Blood samples were obtained at -20, 0, 5, 10, 20, 30, 60, 120, and 180 minutes via catheter, and rLH and rabbit follicle-stimulating hormone (rFSH) levels were determined by RIA. Doses between 1 and 100 IU of hLH produced a prompt decrease in rLH (within 5 minutes); the maximal response occurred within 20 to 30 minutes. Calculated as integrated area of response between 0 and 180 minutes, a dose-response relationship existed between 0.5 and 4.0 IU of hLH. Human LH, even at high doses of 50 IU, produced no changes in endogeneous rFSH. This is the first description of an entirely specific control system for LH, separate from FSH. The short-loop feedback control system for LH is sensitive to levels of LH estimated to be present in eugonadal animals.


Role of blood and fibrinogen in development of intraperitoneal adhesions in rats. One hundred and twenty rats were divided into six groups and laparotomized. In three of the groups the cecum was left intact. In one of these groups the cecum was only lifted up through the abdominal incision and then replaced in the abdominal cavity. In another group the intact cecum was bathed with a certain amount of blood from the animal's tail. In the third group the uninjured cecum was covered with fibrinogen in an amount equivalent to that in the blood used in the other group. In the remaining three groups the cecum was traumatized in a standardized way. In one of these groups the injured cecum was left untreated. In another group the traumatized cecum was bathed in homologous blood and in the third group the injured cecum was treated with fibrinogen as described above. Two weeks later the rats were killed and the number and quality of the intra-abdominal adhesions were studied. The findings in the present investigation indicate that neither blood nor fibrinogen per se induced adhesions to the serosa. On the contrary, a defect in the serosa initiated the formation of adhesions.


[Secretory cycle of pancreatic acinar cells in different functional states]. Time parameter and secretory cycle in acinar cells of the pancreas were studied after atropine, trazilol, pilocarpine administration, and subtotal resection of the small intestine. The duration of secretory cycle was found to be about 45 min. in intact rats. Atropine and trazilol suppressed the secretion process, first inhibiting the release of the formed product and then reducing the synthesis. Pilocarpine first accelerated but very soon decreased the synthetic process. A marked acceleration of synthesis and intensification of secretory cycle in glandular cells occurred in postresectional period. Interrelationships of intracellular mechanisms for the product release and synthesis of new protein molecules, are discussed.


[Cytophysiology of the absorption cycle of enterocytes]. The protein absorption (albumin and gamma-globulin) by epithelial cells of small intestine of neonatal rats was studied in order to define the duration of its stages, the role of intracellular structures in trensport of the absorbed substances, and the cyclic activity of enterocytes. The volume of the absorbed substances depends on the number and size of pynocytotic formations at the cell apex. The protein transport via cells into interstitia is an active process involving the Golgi complex, mitochondria, and other cellular structures. Due to be intracellular regeneration, the restoration of enterocytes begins from gradual protein unloading and is over when the product has been transported to the lymphatic capillary lumen from interstitia. As a result, the continuous absorption is possible in the mucous layer of small intestine although the villi enterocytes activity is of a cyclic character.


[Effect of histamine and pentagastrin on the functional state of the hypothalamo-neurohypophyseal secretory system]. Histochemistry, cario- and zytometry methods showed that the activators of gastric secretion (histamine and pentagastrine) exerted opposite effects upon functional state of hypothalamo-neurohypophyseal secretory system in the guinea-pigs. Histamine (4 mg/kg) activated the system and induced the development of gastric mucosa defects. Pentagastrine (5, 12, 15, 25, and 60 mcg/kg) decreased the activity of the system. No gastric mucosa or duodenum defects developed after pentagastrine administration.


[Anabolic effects of parenterally administered gastric gland hydrolases]. In rats, i.v. administration of amylase, pepsinogen, and tripsinogen in microdoses increased the incretion of radiomethyonin from the blood into the tissue proteins of some organs. The incretion of pepsinogen stimulated the protein-production function of pancreas, tripsinogen stimulated the function of stomach glands, amylase--the function of the liver and the small intestine mucosa. The labeled pepsinogen 125I concentrated mainly in secretory organs.


[Ultrastructure of pancreatic acinar cells under the influence of pancreozymin, serotonin and histamine]. The pancreas was removed from male white mice given intraperitoneally pancreozymin (2 i. u./kg and 4 i. u./kg), serotonin (0.2 and 0.4 mg/kg) or histamine (0.1 mg/kg) within 5--10 min. Pancreozymin caused hypertrophy of the Golgi zone and the appearance of multivesicular bodies. Serotonin induced an intensive autophagia in the apical parts of acinar cells. The effect of histamine was revealed in the rapidity of the extrusion of zymogen granules and the dilatation of cisterns of granular cytoplasmatic reticulum. Possible physiological significance of the ultrastructural alterations of acinar cells is discussed.


[Studies on the purification and the immunological properties of LATS-immunoglobulin (author's transl]. It has been shown that LATS activity is mainly distributed in the fraction of immunoglobulin G (IgG) in the serum from hyperthyroid patients. The present paper examined the immunological character of LATS and the method for separation of LATS activity from LATS positive serum using DEAE-Cellulose and affinity chromatography methods. LATS activity was distributed in the IgG fraction that could be separated by the DEAE-Cellulose column equilibrated with a 0.0175 M prosphate buffer, pH 6.3 from LATS positive serum. When LATS positive serum was fractionated by affinity chromatography on a Sepharose-bound antibody against human IgG, Fab of IgG and Fc of IgG, LATS activity was always retained in IgG fraction. When LATS positive serum was fractionated by affinity chromatography on a Sepharose-bound anti-K chain, LATS activity was found in the fraction that reacted with the anti-K chain. Because of the low antibody titer of the anti-lambda chain, LATS activity did not react with this antibody. By affinity chromatography on Sepharose-bound Concanavalin A, serum LATS activity was also retained in IgG fraction. LATS activity could be separated from LATS positive serum without significant loss of biological activity by affinity chromatography. When IgG (1) was purified from the fraction by affinity chromatography on anti-IgG (1)-bound Sepharose, about 80% of the original LATS activity was found in IgG (1) fraction. When the Fab fragment of IgG (1) was separated from papain hydrolysed IgG (1), using a Protein A-bound Sepharose column, a short-acting type of thyroid stimulating activity was found in only this fraction. These data suggest that the biological activity of the thyroid stimulation is distributed mainly in the Fab fragment of IgG (1).


[A rapid and direct 125 I-based radioimmunoassay for unconjugated estriol in pregnancy serum and its clinical application (author's transl)]. A rapid and simple R.I.A. method for serum unconjugated estriol in pregnancy had been studied for its usefulness as a routine obstetrical test to assess fetal well-being. Diluted serum samples or known amounts of standard estriol were applied with 125 I-estriol on pre-packed minicolumns of Sephadex G-25, followed by appropriately diluted antiserum. During incubation, the columns were counted in an Auto Well gamma-System to read the total counts (T). After a 90 minute incubation period, the columns were eluted with a buffer solution to remove the bound fraction, and the free estriol remaining in the columns was counted (F). Two quality control samples and sera from 74 pregnant women were analysed by this new method, and the results were compared to those obtained by the conventional radioimmunoassay method. The new column method is simple, specific and accurate. Results can be calculated within 4 hours after 30 sample sera are brought into the laboratory. Within and between assay coefficients, variations are 12.6 and 13.5%, respectively. This method correlates well with the results of the conventional method (r=0.7088, p less than 0.001), which requires almost 2 full days to process the same numbers of samples. Results of clinical cases as well as advantages of measuring free-circulating estriol in pregnancy are discussed.


[Temporal relationship of LH and estradiol at mid cycle (author's transl)]. In order to quantitate the feedback effect of estradiol (E2) on the release of LH, serum LH and E2 were measured in 27 ovulatory women daily during the menstrual cycle. The initial rise of LH levels concomitantly occurred when serum E2 concentrations over 100 pg/ml and over 200 pg/ml were maintained for 2 approximately 4 days and for 1 approximately 2 days respectively. The initial rises of LH levels were observed on the day of preovulatory E2 peak. A more detail analysis of the relationship between the two hormones were obtained in five subjects whose blood samples were taken at every eight hours around the time of ovulation. In one among five cases, samples were not obtained early enough to ascertain the relation of the E2 peak to the LH peak. In 3 out of the 4 cases, the intial rises of LH levels occured prior to or concomitant with the time of the E2 peak. By average of all of the five cases, a significant initial rise of LH levels was observed simultaneously with the time of the E2 peak. These results suggest that the increase of the circulatory E2 levels is essential for the initiation of LH surge and their decrease levels are an unrequired mechanism for the midcycle LH release.


[Changes in serum thyrotropin, thyroxine and triiodothyronine after complete thyroidectomy (author's transl)]. Changes in serum TSH, T4 and T3 concentrations after complete thyroidectomy were observed for 4 weeks in 9 euthyroid patients with thyroid carcinoma. Completeness of the thyroidectomy was confirmed by a 131 I scintigram performed one or two months after the operation. Serum TSH levels rose progressively during the 4 weeks of observation, and a significant increase was noticed as early as 3 days after the thyroidectomy. Serum T4 and T3 concentrations decreased significantly 3 days and 18 hrs, respectively, after the thyroidectomy. A sharp decrease in serum T3 concentrations within 18 hrs without a significant change in serum T4 levels and possibly without a significant change in the amount of T3 derived from T4 suggests that the amount of T3 secreted from the thyroid is large enough to affect serum T3 concentrations. The rate of decrease of serum T4 (t 1/2: 16 days) or serum T3 (t 1/2: 23 days) after the 3rd day of the thyroidectomy was much slower than the rate of disappearance of labeled T4 or T3 reported previously. The slow decrease rate of serum T3 is probably due to the conversion of T4 to T3 in peripheral tissues, and that of serum T4 may be due to either the decrease in T4 disposal rate in hypothyroidism or due to the release of T4 from peripheral tissues to serum.


[Microdetermination of corticosteroids in adrenocortical zones in various adrenal diseases (author's transl)]. Since the relationship between steroidogenic activity and morphological differentiation is not necessarily clear in the human adrenal cortex, aldosterone (Al), cortisol (F) and corticosterone (b) from the adrenal tissues cut into four small pieces (capsule and glomerulosa, outer fascicular, inner fascicular, and reticular zones) on a cryostat immediately after their removal at surgery were estimated by radioimmunoassay in 15 patients with adrenal diseases (8 primary aldosteronism, 1 idiopathic aldosteronism, 4 Cushing's syndrome and 2 pheochromocytoma) and 8 control subjects with other diseases. A larger amount of Al was contained in the outer fascicular zone than in other zones in control subjects. Al showed a high value of 0.16 approximately 7.40 ng/mg tissue in adenomas of primary aldosteronism and a low value of 0 approximately 0.25 ng/mg tissue in adenomas of Cushing's syndrome. A high value of Al was detected in idiopathic aldosteronism but not in the remaining adrenal of primary aldosteronism. In Cushing's syndrome, F showed a high value of 2.58 approximately 12.3 ng/mg tissue in adenoma and a relatively low level of 0.77 ng/mg tissue in carcinoma. A larger amount of F and B was found in the inner fascicular zone than in other zones in the control subjects. These results indicate that the corticosteroid content in each adrenal zone correlates with the morphological findings.


Fine structure of the secretion granules in the mandibular gland of the echidna, Tachyglossus aculeatus (Monotremata). The cells of the secretory tubules in the mandibular gland of the echidna are packed with fairly large birefringent granules, which show a lamellated structure consisting of alternating thin and thick layers or shells of protein. This presumably rigid substructure collapses during exocytosis and the shells unravel as sheets that form a tangled mass in the lumen of the secretory tubule. Relatively pure fractions were obtained of the relevant granules and protein sheets, which should allow a further study to be made on the secretory proteins in this gland.


Estrogen and tracheal secretion: the effect of estrogen on the epithelial secretory cells of the rat trachea. Epithelial secretory cells of the tracheal mucosa obtained from ovariectomized control and ovariectomized estrogen-treated rats were studied morphologically by scanning and transmission electron microscopy and by the combined periodic acid Schiff (PAS) and alcian blue (AB) staining method. A significant decrease was observed in the number of large PAS-reactive secretory cells following estrogen treatment. Concomitantly the number of the Mixed+ cells containing both weakly acid and neutral glycoproteins was found to increase significantly. A majority of the observed secretory cells exhibited either an apical protrusion loaded with secretory granules, extending to a maximum distance of 7.9 micrometer into the tracheal lumen or an apical indentation devoid of mature secretory granules, descending to 5.6 micrometer below the luminal surface. The frequency of cells exhibiting an apical indentation was significantly greater in estrogen-treated animals than in controls. The present findings indicate that estrogen treatment of ovariectomized rats results in: 1) an increase in the synthesis of weakly acid glycoprotein within tracheal secretory cells; 2) a concomitant decrease in the frequency of secretory cells containing neutral glycoprotein; and 3) a general increase in the secretion by the tracheal cells.


The influence of microinjected phalloidin on locomotion, protoplasmic streaming and cytoplasmic organization in Amoeba proteus and Physarum polycephalum. Microinjected phalloidin induces both time and concentration-dependent changes in morphology and motility of amoebae and acellular slime moulds. In A. proteus injection of a 10(-3)M solution of the drug causes a separation of cortical hyaline plasma from central granular plasma. Simultaneously protoplasmic streaming and cellular locomotion are lost irreversibly. Lowering the concentration of phalloidin to 2 x 10(-4)M results in a reversible disturbance; amoebae recover after 30 to 60 minutes and show normal movement. In Ph. polycephalum the injection of a 10(-3)M solution of phalloidin into single veins induces a local gelation of the protoplasm followed by the separation of hyalo- and granuloplasm. In semi-thin and ultrathin sections the hyaline plasma regions contain a fine granular groundplasm rich in ribosomes but free of cellular organelles. The central granular plasma consists mainly of membrane-surrounded cellular compartments. The two morphologically distinct plasma regions are separated by a 0.5 to 1.0 micrometer layer of filamentous material. In A. proteus the filamentous layer is found shortly after phalloidin injection in close proximity to the plasma membrane, and consists of thin 5 to 6 nm filaments. With increasing time this layer contracts, separates from the inner plasma membrane and moves to the interior of the cell. During contraction thicker filaments with diameters of 10 to 30 nm and lengths of 300 to 500 nm are formed. The results indicate that the display and contraction of the phalloidin-induced filament layer can account for the changes observed in cellular movement and cytoplasmic organization. The resulting phenomena i.e. separation of hyaline plasma from granular plasma and changes in both the protoplasmic streaming pattern and locomotory activity of the cells, are discussed in terms of a general understanding of amoeboid movement.


Chromatin activation in peritoneal exudate leukocytes after fusion with L cell cytoplasts. Cells reconstituted from the L cell cytoplasts and nuclei of mouse peritoneal leukocytes (PL) have been obtained by fusion procedures. The dormant nuclei of leukocytes undergo reactivation after fusion with active L cell cytoplasms. The reactivation is manifested by the swelling of nuclei, the increase of 3H-actinomycin D binding to chromatin in situ and the 3H-uridine incorporation in nuclei in vivo and by the appearance of nucleoli. The data obtained indicate that the process of dormant nuclei reactivation does not require the presence of the nucleus of the active cell partner. Moreover, the process of chromatin reactivation is more rapid in reconstituted cells than in PL + L cell heterokaryons. Some other details of PL chromatin activation in reconstituted cells and heterokaryons are discussed.


Reaction of the contractile apparatus in Physarum to injected Ca++, ATP, ADP and 5'AMP. After replacement of endoplasm by artificial media, the ectoplasmic tube of the plasmodial veins survives and contraction automaticity continues. Motive force generation for plasmodial locomotion is widely independent of the presence of endoplasm, i.e. force generation is located within the ectoplasmic tube. The replacement of endoplasm offers the possibility to study the effects of physiologically active substances upon the contraction activity of cytoplasmic actomyosin under in vivo conditions. Using different methods for measuring the force output as indicator of the effects of injected substances, the influences of free Ca++-ions and different nucleotides were studied. The injection of Ca++ buffers revealed that force output of the veins increased with ascending Ca++ concentrations between 2 x 10(-7) and 10(-6) M. Half maximal response was found to lie in the range of 4 x 10(-7) M, the saturation value at 6 x 10(-7) M Ca++. The injection of adenosine nucleotides induced optimal contraction responses at 0.2 mM ATP, 0.5 mM ADP and 2.5 mM 5'AMP, respectively. The contraction response induced by 0.2 mM ATP depends strongly on the concentration of the available Ca++. From a threshold concentration of 2 x 10(-7) M Ca++, force output increased up to 10(-5) M Ca++. In presence of 0.2 mM ATP, the value for half maximal contraction response to Ca++ was 10(-6) M Ca++. It is concluded that 0.2 mM ATP and 2 to 4 x 10(-7) M Ca++, are the conditions, under which the cytoplasmic actomyosin system of Physarum performs its function in vivo. The implication of the results and the possibilites of the new method for further investigations of the contraction physiology of Physarum as a model system for cytoplasmic actomyosin are discussed.


Glucose tolerance in jejunoileal bypass for morbid obesity: a fifteen month follow-up. Twelve patients with body weight varying from 106 to 163 Kg, underwent jejunoileal bypass operations; their oral glucose tolerance tests (oGTT) were evaluated before the operation and one, five, and fifteen months afterwards. The lowering of the glycemic curve and the decrease of the insulinemia values during oGTT have been studied in relation to intestinal absorption deficit and weight loss. It is concluded that weight loss was the main factor determining the improvement of glucose metabolism.


Humoral antibacterial immunity in first degree relatives of insulin-dependent diabetics. Humoral immunity to bacterial antigens was investigated in 68 tissue typed and glucose tolerance tested first degree blood relatives of insulin dependent diabetics (IDD). The data were compared with those obtained in 60 IDDs and in 55 healthy controls. The prevalence of bacterial antibodies to E. coli, staphylococci, pertussis and diphtheria toxins were just slightly, but not significantly reduced in the blood relations compared with controls. Incidence of antibacterial antibodies was almost identical in blood relations with impaired and in those with normal glucose tolerance. By contrast, antibody formation to E. coli and staphylococci (p less than 0,0005, p less than 0,0005) respectively was significantly impaired in IDD. No correlation between genes of the major histocompatibility complex and humoral antibacterial immunity could be observed in IDD and blood relations. In conclusion, antibacterial antibody formation was found to be severely impaired in IDD patients but to be almost normal in blood relations of insulin dependent diabetics. These findings suggest that the humoral antibacterial immunodeficiency observed in IDD is a disease associated process probably independent of major histocompatibility complex linked genes.


[Adipocyte size in primary hypertriglyceridemia with and without obesity (author's transl)]. Adipose tissue cellularity of patients with endogenous hypertriglyceridemia whether associated with obesity or not has been studied by means of a semiautomatic method of counting and sizing osmium tetroxide fixed adipocytes. Cell population distributions were analyzed by mathematical computation. The Coulter Counter system was able to determine mean diameter (or volume) and cell size dispersion which defined adipocyte population. An overall shift of cell population size has been found to characterize adipose tissue cellularity of the different groups examined. Adipocyte population in the obese was defined by increased fat cell size with overlapping distributions between both groups of same weight. Nevertheless normolipemic obese patients were characterized by larger cell size than hypertriglyceridemic subjects matched for adiposity index. The results are discussed in relation to the removal defect found in endogenous hypertriglyceridemia.


Effects of prostaglandin E1 and prostaglandin F2alpha on insulin and glucagon plasma levels during the intravenous glucose tolerance test in man. The influence of prostaglandin E1 and prostaglandin F2alpha on circulating concentrations of insulin and glucagon during the intravenous glucose tolerance test has been studied in normal man. Insulin responses to glucose during PGE1 infusion (0.2 microgram/Kg/min) were significantly lower than in control infusions (p less than 0.001 at 2, 5, 10 and 15 min). Moreover, PGE1 caused a clear elevation of basal glucagon (p less than 0.02). PGF2alpha, at the two doses used (0.2 and 0.5 microgram/Kg/min), had no effect on basal glucose, insulin and glucagon levels, nor upon glucose-induced insulin secretion. Neither prostaglandin affected the glucose-induced inhibition of pancreatic alpha-cells. There is thus some evidence that PGs of E series may play some role in modulating the secretion of human pancreatic beta-cells.


Ascorbic acid status in idiopathic hemochromatosis. Vitamin C status was studied, by means of leucocyte ascorbic acid concentrations, in 67 cases of idiopathic hemochromatosis subdivided into 44 untreated and 25 treated cases (2 patients belonging to both subgroups) and compared to 31 normal subjects and 37 alcoholic cirrhosis patients. The control groups exhibited the following mean levels (+/- SEM): 34.4 +/- 1.9 microgram/10(8) WBC in normals and 22.0 +/- 1.8 microgram/10(8) WBC in alcoholic cirrhosis. In idiopathic hemochromatosis the mean levels were: for the untreated group 19.5 +/- 1.7 microgram/10(8) WBC and for the treated group 34.3 +/- 2.3 microgram/10(8) WBC. These results (1) affirm an important vitamin C deficiency in the untreated disease; (2) suggest that iron overload is the main causal factor in view of the striking difference--to date unreported--between untreated and treated cases of idiopathic hemochromatosis. Besides its possible theoretical interests, this vitamin C deficiency is responsible in idiopathic hemochromatosis for a significant underestimation of the desferrioxamine-induced urinary iron excretion.


Fine structural changes in the liver of methotrexate-treated psoriatics. Ultrastructure of 55 liver biopsies obtained from 52 patients, all but 1 with psoriasis and all but 5 treated with methotrexate (MTX), have been studied. In MTX-treated psoriatics, liver cell nuclei showed reduced electron opacity with frequent cytoplasmic invaginations and glycogen inclusions. In the mitochondria nonspecific changes, such as pleomorphism, gigantism, paracrystalline inclusions, compartmentalization, signs of division and sequestration via autophagosomy were noted. The pericanalicular ectoplasm exhibited widening with accumulation of microfilaments. Detachment of desmosomal plaques between hepatocytes was common. A striking finding was the hyperplasia of fat-storing perisinusoidal (Ito) cells. No definite correlation was found between morphology, duration of disease, treatment as well as cumulative dose of MTX.


Physiologic release of secretin measured in peripheral and portal venous blood of dogs. The effect of food on circulating levels of secretin, measured by radioimmunoassay, was studied in 7 dogs. Significant postprandial increases in secretin were found in portal and peripheral plasma of dogs fasted for 18 h. Basal levels of secretin were significantly lower in dogs fasted for 72 h when compared with dogs after an 18-hour fast. After the prolonged fast, food caused a significant release of secretin, measured peripherally. We conclude that secretin is released by a physiologic stimulus (food) and that circulating levels of secretin are depressed after prolonged starvation.


Lack of parallelism between microsomal enzyme induction and phenobarbital-induced hypercholeresis in the rat. The relationship between microsomal enzyme induction and the increase in bile flow associated with phenobarbital administration was studied in rats in three experimental situations: examination of the time-course effect of a single dose of phenobarbital (8 mg/100 g body weight) on bile flow and hepatic cytochrome P-450 concentration; study of the influence of SKF 525-A (8 mg/100 g body weight) and cobaltous chloride (6 mg/100 g body weight/day for 3 days) on the phenobarbital-induced hypercholeresis. It was observed that: (a) the maximal increase in bile flow occurred 18 h after the single injection of phenobarbital, while the maximal increase in cytochrome P-450 occurred at 48 h; (b) in rats pretreated with phenobarbital for 3 days, SKF 525-A did not suppress the hypercholeresis due to phenobarbital, and (c) in rats treated with phenobarbital and cobaltous chloride, cytochrome P-450 concentration in the liver was not increased, while bile flow was increased to approximately the same extent as in animals treated with phenobarbital alone. These results further support the hypothesis that microsomal cytochrome P-450-dependent enzyme induction and increase in bile flow are two separate effects of phenobarbital.


Antral vagotomy in parietal cell vagotomized dogs with a Heineke-Mikulicz pyloroplasty. Effect upon gastric emptying and motility, serum gastrin concentration, and Heidenhain pouch acid secretion. In 6 Heidenhain pouch (HP) dogs who previously had undergone parietal cell vagotomy (PCV) with a Heineke-Mikulicz pyloroplasty (Ppl), the antral nerves were cut. No significant changes in HP acid secretion or serum gastrin concentration occurred. In a similar previous study using HP dogs with a PCV and a gastroduodenostomy (GD), a significant rise in HP acid secretion without concomitant changes in serum gastrin concentration were observed after antral denervation. These findings indicate that the antral vagal nerves have no direct influence on HP acid secretion, but the rised HP acid secretion in the GD dogs after antral denervation may be related to the changed gastroduodenal emptying pattern which partly excludes the food acid stream from the acid inhibitory mechanism in the duodenal bulb.


Effects of prostaglandins and their methylated analogues upon human adenylate cyclase in the upper gastrointestinal tract. The effects of prostaglandin E2, prostaglandin A2 and their methylated analogues upon the adenylate cyclase in human gastric mucosa were studied. PG E2 and 16,16-dimethyl-PG E2 stimulated the enzyme activity to a greater degree than PG A2 and 16,16-dimethyl-PG A2. 16,16-dimethyl-PG E2 was less potent in activating the enzyme system than its parent compound. The results suggest that the more pronounced antisecretory activity of 16,16-dimethyl-PG E2 is due to its greater resistance against enzymatic degradation.


Effect of food intake on circulating antigen-antibody complexes in patients with alcoholic liver cirrhosis. Serum antibodies directed against 5 alimentary antigens and antigen-antibody complexes were determined in 15 control subjects and in 15 patients with alcoholic liver cirrhosis. Cirrhotic patients presented an increased immune response against these antigens. Antigen-antibody complexes were detected in the fasting plasma of 7 cirrhotic patients. The level of immune complexes increased in 6 out of these 7 patients 1 h after lunch.


Continuous blood glucose analysis in vitro and in vivo. A practical method for continuous blood glucose analysis in vivo is described. Using an interference-free enzyme reagent in a modified Auto-Analyzer whole blood glucose concentration can be monitored continuously and interpreted in terms of the actual plasma glucose concentration. The method uses a novel technique for preheating the sample diluent without introducing additional time delays, consumes whole blood at a rate of 0.05 ml/min, and demonstrates a transport delay of 148 s. Other improvements include an average baseline drift of -0.11 mg/dl/h and a mean change in sensitivity of -0.4% after 8.5 h. In vitro glucose recovery studies comparing whole blood to the corresponding plasma samples show the method is precise (101.3 +/- 0.7%), linearly proportional (slope of 1.007 +/- 0.012) and highly correlated (greater than 0.998) over the range of 0 to 500 mg/dl, with reference to a Beckman glucose analyzer. In vivo applications are presented to show that this method is suitable for us in systems such as the 'artificial endocrine pancreas'.


Thickening of glomerular basement membrane in spontaneously diabetic rats. The glomerular basement membrane of spontaneously diabetic rats was investigated by quantitative analysis using electron microscopy, with special reference to the effect of ageing. Constant age-related increase in the width of basement membrane was ascertained both in diabetic and control rats, and the mean values of basement membrane thickness were always higher in the spontaneously diabetic rats than in normal control rats. Significant thickening of glomerular basement membrane was found in the diabetic rats at 12 weeks of age, while younger diabetic rats had no definite increase. The difference in basement membrane thickness between diabetic and normal control rats became larger with increasing age.


Effect of free fatty acids and amino acids on glucagon and insulin secretions in normal and diabetic ducks. The relationship between two metabolites free fatty acids (FFA) and amino acids (AA), and the two main pancreatic hormones, insulin and glucagon, was studied by infusing small amounts of these metabolites into normal and diabetic Peking ducks, i.e. two days after subtotal pancreatectomy. Infusion of oleic acid (0.365 g/kg/30 min as an emulsion in plasma) indicated a suppressive effect of free fatty acids on glucagon secretion, but was without effect on insulin secretion, in normal as well as in diabetic ducks, indicating that insulin might not be directly involved in the FFA-glucagon feedback in the duck. Infusions of arginine for one hour (1 g/kg/h) into normal ducks, hyperglycaemic normal birds (as a result of glucose infusion: 1 g/kg/h) and diabetic ducks, suggested the persistence of amino acid effect on glucagon secretion, and a slight reduction of the effect on insulin secretion in diabetes. This suggests that insulin may not be involved in amino acid-induced glucagon secretion in the duck.


Sulphydryl requirement for insulin release from the perfused pancreas. Studies with ethacrynic acid and dithiothreitol. Using the isolated, perfused rat pancreas the importance of sulphydryl groups for the secretory process of insulin was investigated. It was found that ethacrynic acid (EA, 0.075-0.6 mmol/1) caused a dose-dependent, monophasic insulin release. Addition of EA to a glucose-stimulated (20 mmol/1) pancreas led to a sudden increase in hormone release, followed by a dose-dependent inhibition of release, which was not reversible after removal of EA. The same phenomenon was seen in the presence of 20 mmol/1 leucine. Dithiothreitol (DTT, 0.1 and 1 mmol/1) had no effect on basal insulin secretion. Added to a glucose-stimulated pancreas DTT (1 mmol/1) caused a reversible inhibition of insulin release. The persistent inhibitory action of EA on glucose-induced insulin release could be reversed by simultaneous perfusion of EA and DTT. Sequential exposure of a glucose-stimulated pancreas to EA and DTT led to a rapid release of insulin, due to DTT; however, the EA-induced inhibition of insulin secretion could not be prevented. Two kinds of thiol groups in the plasma membrane and in the beta cell might be responsible for the various kinetics of insulin release induced by EA and DTT.


Haloperidol, a dopaminergic antagonist: somatostatin-like inhibition of glucagon and insulin release from the isolated, perfused canine pancreas. The effect of haloperidol, a dopaminergic antagonist, on insulin and glucagon secretion was investigated using the isolated, perfused canine pancreas. Haloperidol at 4 X 10(-7) to 10(-5) mol/l caused a dose-dependent inhibition of glucagon release both at low (25 mg/100 ml) and high glucose concentrations (150 mg/100 ml). At the low glucose concentration insulin release was already maximally suppressed. At the high glucose concentration haloperidol (4 X 10(-7) to 10(-5) mol/1) also caused a dose-dependent inhibition of insulin release. Haloperidol (10(-5) mol/1) inhibited dramatically pancreatic A and B cell responses to isoproterenol (2 ng/ml), acetylcholine (1 mumol/1) and arginine (5 mmol/1). The inhibitory effect of haloperidol on both glucagon and insulin release could be eliminated by increasing perfusate calcium concentration from 1.3 to 8.8 mmol/1. These findings suggested that haloperidol blocks glucagon and insulin release in a somatostatin-like manner by affecting a fundamental step of the stimulus-secretion coupling, probably by interfering with calcium handling of the pancreatic A and B cells.


[Clinical use of a minicomputer in heart surgery intensive care units]. The Authors present three respiratory parameters useful to estimate the respiratory and hemodynamic conditions of a cardiac patients before these turn into obvious pathological clinical pictures. The parameters are: alveolar-arterial oxygen difference, respiratory index and arterial-venous pulmonary shunts. The sophisticated mathematic calculations necessary to calculate and to elaborate such data are made, in few seconds, by a minicomputer which is very easy to work with. A clinical example in which the above-mentioned data have been employed helps to demonstrate their extreme usefulness.


[The administration of an association of sodium nitroprusside and adrenaline in cardiac surgery (author's transl)]. In 223 operations of cardiovascular surgery, myocardial performance was supported, after the interruption of cardiopulmonary bypass, by means of a combined and strictly controlled administration of a pure vasodilator drug (sodium nitroprusside) and of an inotropic agent (epinephrine). The reduction of afterload achieved by the former, the increase of contractility and heart rate induced by the latter, the coronary dilator effect of both, associated with an adequate maintaining of preload proved to be rapidly effective. This appeared to be particularly true in patients with overt left ventricular failure after valve replacement and/or myocardial revascularization procedures. The proposed association of drugs and treatment program seem to be an appropriate way of managing the myocardial and hemodynamic response to the metabolic reactivation that follows the surgical ischemic cardiac arrest.


["T'orsade de pointe" and amiodarone (author's transl)]. A long Q-T interval syndrome is described, followed by "torsade de pointe" and by irriducible ventricular fibrillation that is ascribed to a badly conducted therapy with amiodarone, in a patient affected by mitral valve disease, microcitaemia and hemolitic intercurrent moderate jaundice.


[Natural history of angina pectoris: follow-up on 519 unoperated patients (author's transl)]. 519 patients with angina pectoris studied by selective coronary arteriography and left ventriculogram, were followed for a period ranging from 18 months to 7 years. The mean follow-up was 42.2 months. The patients showed a survival probability of 81% at the 7th year. After 5 years the survival probability was 83.2% for patients with typical stable angina, 70.3% for patients with unstable angina, 96.7% for patients with atypical angina. The survival probability was 78.8% for the male sex and 94.6% for the female (at the 5th year). Age, a long-lasting angina, the presence of: previous infarction, myocardial failure, cigarette smoking, hyperlipidemia, cardiomegaly and an ischemic resting EKG were factors with poor prognostic value. The prognostic value of significant coronary stenosis was confirmed. The survival probability at the 5th year of the patients without critical stenosis was 96.6%, of patients with stenosis of 1, 2 and 3 main coronary arteries was respectively: 87.6%, 79% 54.7%. Significative prognostic differences were observed in patients with normal left ventricle kinesia (survival probability at the 5th year: 90%), compared with patients with severe VS ipokinesia (62.7%) and with VS diskinesia (69%). In the follow-up period an incidence of 9% of myocardial infarctions was observed. The degree of each stenosis and the number of vessels involved, the type of angina, the presence of risk factors or previous myocardial infarction did not affect the clinical evolution of angina.


[Myocardial infarction in young people. A coronary angiographic study in 21 patients under the age of 30 (author's transl)]. The Authors have taken into account 21 patients who complained of myocardial infarction before they were 30 years old. We performed coronary angiography, left ventriculography and left cardiac catheterization in all the patients. In 20 cases we found atherosclerotic alterations (50 or greater obstruction) at least in one of the three main braches of  coronary tree. The coronary angiography in one patient showed a picture of dissection of right coronary. The alterations are localized more frequently on the anterior descending branch than on the right coronary. On the contrary we found very seldom atherosclerotic lesions on the circumflex artery. Monovascular alterations were more frequent than plurivascular alterations (more than 40% of the total). Left ventricular function was out of order in almost all cases. In summary the important differences between young patients with myocardial infarction and older patients are the following: 1) from the clinical point of view the sudden onset of myocardial infarction without previous symptoms of angina pectoris; 2) from the angiography point of view high incidence of monovascular alterations and low incidence of collateral circulations.


[An echocardiographic study of the isovolumetric relaxation period in the myocardial infarction (author's transl)]. Variations have been examined in the left ventricle's transverse dimension (TD) during the isovolumic relaxation period (IRP) in 12 healthy subjects and in 38 patients with anterior or antero-inferior infarct. Healthy subjects have shown no increase in TD during the IRP whereas this fact has been noted in 24 out of 38 (63%) patients with different distribution according to the site of the infarct (29%) for inferior infarcts and 86% for anterior or antero-inferior infarcts). These results suggest that inferior infarct compared with anterior infarct, implies a less incoordinate ventricular contraction. It is thus stressed the opportunity of echocardiography approach for the identification of the incoordinate ventricular contraction and for the determination of IRP duration which may be exposed to serious mistakes using the traditional phono-apexcardiographic method.


[Usefulness of x-ray parameters in the functional evaluation of patients with nonvalvular cardiopathies]. In 49 patients suffering of heart diseases we have studied the changes of radiologic cardiac measurements and systolic time intervals (STI) in the four functional classes of the New York Heart Association (NYHA) classification, investigating also the existence of any relationship between these different parameters. Only the patients in functional classes 3rd and 4th showed significant changes in STI and radiologic measurements as compared to the control group. Moreover, a significant negative correlation has been observed between relative heart volume and left ventricular ejection time (LVET) (r = 0.69, P less than 0.001) and LVETc (r = 0.82, P less than 0.001) and a positive correlation between relative heart volume and pre-ejection period (PEP) (r = 0.59, P less than 0.01) and PEP/LVET ratio (r = 0.75, P less than 0.001). These results seem to demonstrate that relative heart volumetry is a fairly accurate index of the cardiac conditions in non valvular heart diseases.


[A specific anomalies of ventricular repolarization. Dysgenetic syndromes caused by functional asymmetry of the cardiac sympathetic nerves]. The following is a personal study of the case histories of 3 patients, spontaneous carriers of "labile pseudo-ischemic T wave" and of their direct consanguineous, a total of 25 subjects. In these three families the family incidence on electrocardiographic anomalies is confirmed. In these same subjects as well as in others belonging to families examined in a previous study, a total of 90 subjects, the duration of the QT interval was measured, showing values which tended to be above the average and, in some cases, above the accepted maximum. The analogy among the anomalies of the ventricular repolarization to be found during the course of neurological diseases and under experimental conditions of unilateral lesions of the sympathetic cardiac innervation and the "labile T wave" syndrome, sometimes coexisting in the same family with a report of elongated QT interval, suggests the hypothesis that this may be placed within the ambit of disgenetic syndromes from alterations of the functional balance between right and left component of the sympathetic heart innervation.


[Reflex pressor response in young subjects with labile hypertension (author's transl)]. A study on the pressor response to various tests of sympathetic stimulation has been performed in 36 young subjects, 9 with labile hypertension and 27 normotensive. All have been studied with "tilt test", "cold pressor test" and "mental arithmetic stress". In the labile hypertensive subjects the "tilt test" induced increases of the systolic and diastolic pressure significantly higher than in the normotensives. The "cold pressor test" produced an increase of the arterial pressure and of the heart rate significantly lower in the labile hypertensive than in the normotensive. The pressure responses and the changes of the heart rate produced by the "mental arithmetic stress" did not show significant differences in the two groups of patients studied. The results are discussed.


Thromboplastic and fibrinolytic activities of ascites tumor cells of rats, with reference to their role in metastasis formation. Thromboplastic and fibrinolytic activities of rat ascites tumor cells avoiding any stromal elements were examined and the role of these activities in the blood-borne metastasis was discussed. Ten lines of tumor cells showed varied thromboplastic and fibrinolytic activities. Tumor cell lines examined were classified into four groups; (1) lines AH-130, AH-62, and AH-7974 with high thromboplastic and high fibrinolytic activities, (2) lines AH-130F(N), AH-66F, and AH-7974F with low thromboplastic and low fibrinolytic activities, (3) line SLC with high thromboplastic and low fibrinolytic activities, and (4) lines AH-109A, AH-41A, and AH-41C with moderate thromboplastic and low fibrinolytic activities. The cell lines AH-130 and AH-130F(N), as well as AH-7974 and AH-7974F, have the same origin and showed different enzymic activities. AH-130 caused more prominent thrombus formation in the pulmonary vessels of rats in the early stage of intravenous inoculation and induced more prominent decrease in the number of platelets and fibrinogen levels in peripheral blood than AH-130F(N). Also, AH-130 developed more abundant metastatic foci in the lung 72 hr and 7 days after intravenous inoculation than AH-130F(N).


Autoradiography and organ distribution of N-methyl-N-nitro-sobenzylamine in rats. Donryu strain male rats were administered a large amount of N-methyl-N-nitrosobenzylamine directly into the stomach through a tube. Ulcers were found to develop in the portion of the esophagus where esophageal carcinoma is known to develop. Autoradiography of the entire body of rats after an intravenous injection of N-methyl[U-3H]-N-nitrosobenzylamine showed specific incorporation of radioactivity in the esophagus. When N-methyl[U-3H]-N-nitrosobenzyl-amine was injected intravenously, incorporation of radioactivity in the esophagus was higher than in other parts of the alimentary canal, and remained at the same value even after 24 hr. Microautoradiography of rats injected with N-methyl[U-3H]-N-nitrosobenzylamine through the tail vein showed that the silver grains were dominant in the mucous membrane than in the muscle layer in the esophagus. These results suggest that N-methyl-N-nitrosobenzylamine accumulated specifically in the esophagus, resulting in the production of carcinoma.


Comparative immunological studies on cryosurgery and surgical operation using Moloney murine sarcoma virus-induced primary tumors in BALB/c mice. The effect of cryosurgery and surgical operation on Moloney murine sarcoma virus-induced primary tumors in the treated and untreated groups of BALB/c mice was compared in terms of tumor growth, cumulative mortality, lymphocyte-mediated cytotoxicity, proliferative response of lymphocytes, and humoral antibody formation. The results indicate that cryosurgical treatment showed both in vivo and in vitro effects; (i) tumor growth and cumulative mortality in mice treated by cryosurgery were significantly lower than those in untreated groups, and (ii) their cellular immune response was enhanced, as manifested by increase in a proliferative response and in lymphocyte-mediated cytotoxic activity against Moloney murine leukemia virus-induced lymphoma. The proliferative response of spleen cells and the cytotoxic activity of lymphocytes were not parallel; the proliferative response detected by 3H-thymidine incoroporation manifested peak activity 3 days after cryosurgery, but the cytotoxic activity detected by 125I-iododeoxyuridine-releasing tests was considerably decreased at this stage. Two weeks after cryosurgery and thereafter, however, the cytotoxic activity of this group increased to a level higher than that of untreated or surgically treated groups. On the contrary, surgical operation abrogated the proliferative response of spleen cells. Immunofluorescence tests revealed, however, that humoral antibody formation was higher in surgically operated groups than in other groups.