[Actions of sodium dichloroacetate in combination with insulin on hyperlactatemia and hyperpyruvicemia induced in dogs by phenformin]. In the normal anesthetized dog the combination of insulin, whether of exogenous or endogenous origin, with sodium dichloroacetate provoke a rapid and important reduction of the hyperlactatemia and hyperpyruvicemia induced by the intraduodenal injection of high doses of phenformin. Furthermore this combination prevents the progressive and important lowering of the arterial pH provoked by phenformin.


[Effects of a beta-adrenolytic and a diuretic vis-à-vis hyper-reninemia induced by isoprenaline in anesthetized dogs. Value of beta blocking-diuretic interaction]. Intravenous injection of isoproterenol increases plasma renin activity (PRA) in anesthetized dogs. S. 464, a new beta adrenergic blocking agent, injected five minutes before isoproterenol, inhibits plasma renin hyperactivity. On the other hand, teclothiazide, a thiazide diuretic, induces no significant modification of the isoproterenol-induced increase of PRA. The combination of both compounds (five parts of S. 464, one part of diuretic), assumes the same inhibitory effects as S. 464 alone. These results and other experimental data (antihypertensive and diuretic activities) are discussed and explain the interest of such an association as a rational therapy of arterial hypertensive disorders.


[Action of 3-beta adrenolytics on plasma renin activity measured by radioimmunology in genetically hypertensive rats]. Spontaneously hypertensive rats (SHR) have basal levels of plasma renin activity (PRA) lower than the ones observed in normal Sprague Dawley rats. Three beta blocking agents are orally administered to unanesthetized spontaneously hypertensive and normotensive rats. Propranolol and S 464 reduce PRA in spontaneously hypertensive and normotensive control rats. Pindolol do not lower PRA in normotensive rats but increases levels of PRA in spontaneously hypertensive rats. These results are discussed.


[Partial disorganization of the anaphasic segregation of chromosomes in plant cells: combined actions of griseofulvin, producer of pluripolar anaphases and 2 ipecac alkaloids, producers of floating pole anaphases]. Anaphasis may be slightly checked by various treatments which however result in a normal chromosomic separation. Griseofulvin exerts a direct though partial influence on the mitotic apparatus, which entails "pluripolar anaphasis"; on the other hand Ipecac alkaloïds act indirectly and produce "floating poles anaphases". Treatments combining griseofulvin with cepheline or tubulosine show that there is never any synergy between the two processes. These results support our hypothesis that floating poles anaphases are not a sign of slight C-mitotic action but only come from a lag between the appearance/disappearance of microtubules and that of chromosomes during anaphasis.


Peripheral vascular and cardiac effects of nitrous oxide in the bovine. Peripheral vascular and myocardial effects of increasing concentrations of nitrous oxide (0 to 70 per cent) in oxygen were determined in 15 unanaesthetized calves before and after replacement in their natural heart (NH) with a pneumatically driven artificial heart (AH). Nitrous oxide produced concentration-related decreases in arterial and mixed venous pH and increases in minute ventilation and arterial and mixed venous carbon dioxide tensions in both NH and AH calves. Nitrous oxide resulted in significant increases in cardiac output, stroke volume and mean aortic, pulmonary artery and right atrial pressures in NH and AH calves, but did not significantly change systemic vascular resistance in either group of animals. Heart rate was increased in NH calves but was fixed in AH calves. Elevations in heart rate and cardiac output at nitrous oxide concentrations greater than 30 per cent and aortic pressure at 70 per cent nitrous oxide were significantly greater in NH than AH animals (P less than 0.05). These data demonstrate that nitrous oxide stimulates the cardiovascular system in spontaneously breathing mammals and that the changes result from improved venous return and an increase in myocardial chronotropy. Our findings also suggest that cardiovascular stimulation during nitrous oxide breathing may be related to increased concentrations of arterial and/or venous carbon dioxide.


Purification and partial characterization of an exo-beta-glucanase from the yeast Kluyveromyces aestaurii. The intracellular-periplasmic exo-1,3-beta-glucanase (EC 3.2.1.58) has been extracted from the yeast Kluyveromyces aestuarii and purified to immunoelectrophoretic homogeneity by ion-exchange and gel-exclusion chromatography. The kinetic constants and activation energies for laminarin, p-nitrophenyl-beta-D-glucoside, and pustulan have been determined, along with the effect of pH. Evidence is presented indicating that the enzyme is composed of a single polypeptide chain, about 24% carbohydrates, and its molecular weight was estimated to be 43 000.


The role of divalent cations in the activation of the NADP+-specific isocitrate dehydrogenase from Pisum sativum L. A divalent cation electrode was used to measure the stability constants (association constants) for the magnesium and manganese complexes of the substrates for the NADP+-specific isocitrate dehydrogenase (EC 1.1.1.42) from pea stems. At an ionic strength of 26.5 mM and at pH 7.4 the stability constants for the Mg2+-isocitrate and Mg2+-NADP+ complexes were 0.85 +/- 0.2 and 0.43 +/- 0.04 mM-1 respectively and for the Mn2+-isocitrate and Mn2+-NADP+ complexes they were 1.25 +/- 0.07 and 0.75 +/- 0.09 mM-1 respectively. At the same ionic strength but at pH 6.0 the Mg2+-NADPH and Mn2+-NADPH complexes had stability constants of 0.95 +/- 0.23 and 1.79 +/- 0.34 mM-1 respectively. Oxalosuccinate and alpha-ketoglutarate do not form measureable complexes under these conditions. Saturation kinetics of the enzyme with respect to isocitrate and metal ions are consistent with the metal-isocitrate complex being the substrate for the enzyme. NADP+ binds to the enzyme in the free form. Saturation kinetics of NADPH and Mn2+ indicate that the metal-NADPH complex is the substrate in the reverse reaction. In contrast the pig heart enzyme appears to bind free NADPH and Mn2+. A scheme for the reaction mechanism is presented and the difference between the reversibility of the NAD+ and NADP+ enzyme is discussed in relation to the stability of the NADH and NADPH metal complexes.


The interaction of human glycophorin with 8-anilino-1-naphthalene sulfonate. Both the sialoglycoprotein of human erythrocyte membranes, glycophorin, and the sialic acid free protein, obtained by treatment of glycophorin with neuraminidase (EC 3.2.1.18), increase the fluorescence of 8-anilino-1-naphthalene sulfonate (ANS). Binding of ANS to glycophorin is weak compared with the binding to bovine serum albumin (BSA). equilibrium dialysis gives an apparent binding constant of about 4 X 10(3) M(-1) at neutral pH, but Ka increases 1.75 times when NaCl or CaCl2 are added and 10-fold when the pH is lowered to 3.0. Sialic acid groups do not significantly affect ANS binding, although they have some effect at low ionic strength and neutral pH. Fluorescence studies indicate only one to two binding sites for ANS, with apparent pK = 3.8 +/- 0.2, and located close to aromatic residues in glycophorin. Polarization and quantum efficiency of the fluorescence of ANS associated with glycophorin fail to indicate changes in the vicinity of the binding site when the pH is lowered.


Purification and partial characterization of cysteine-glutamate transaminase from rat liver. Cysteine-glutamate transaminase (cysteine aminotransferase; EC 2.6.1.3) has been purified 149-fold to an apparent homogeneity giving a specific activity of 2.09 IU per milligram of protein with an overall yield of 15%. The isolation procedures involve the preliminary separation of a crude rat liver homogenate which was submitted sequentially to ammonium sulfate fractionation, TEAE-cellulose column chromatography, ultrafiltration, and isoelectrofocusing. The final product was homogenous when examined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS). A minimal molecular weight of 83 500 was determined by Sephadex gel chromatography. The molecular weight as estimated by polyacrylamide gel electrophoresis in the presence of SDS was 84 000. The purified enzyme exhibited a pH optimum at 8.2 with cysteine and alpha-ketoglutarate as substrates. The enzyme is inactivated slowly when kept frozen and is completely inactivated if left at room temperature for 1 h. The enzyme does not catalyze the transamination of alpha-methyl-DL-cysteine, which, when present to a final concentration of 10 mM, exhibits a 23.2% inhibition of transamination of 30 mM of cysteine. The mechanism apparently resembles that of aspartate-glutamate transaminase (EC 2.6.1.1) in which the presence of a labile hydrogen on the alpha-carbon in the substrate is one of the strict requirements.


Electrolyte levels and net fluid and electrolyte movements in the gastrointestinal tract of weanling swine. Electrolyte concentrations, osmolality and pH were determined in conventionally raised weanling swine fed a liquid diet. Incorporation of a dilution marker into the diet in combination with frequent feeding enabled estimations as to the sites of relative fluid and electrolyte absorption and secretion along the gastrointestinal tract. Unlike many other species the weanling pig depends largely on its large intestine for absorption of fluid and electrolytes with small changes in net fluid movement occurring along the jejunal and ileal segments. Additional observations included the absorption of water by the porcine stomach which increased dilution marker concentration by approximately twofold and the high osmolality values recorded in the small and large intestine. The implications of these observations are discussed with regard to pathogenesis of colibacillary diarrhea in the weanling pig.


Studies on the antigenicity of an inactivated, aluminum hydroxide adjuvant equine influenza vaccine. An inactivated, aluminum hydroxide adjuvant equine influenza vaccine was tested in horses and guinea pigs to determine the levels of antigen that would elicit maximum serological responses. Vaccine containing serial twofold increments of A/Equi-1/Prague and A/Equi-2/Miami strains of equine influenza virus was administered to random groupings of both types of test animals. The hemagglutination inhibition antibody response for each group was then measured. Results in horses and guinea pigs were compared to determine if the equine serological values could be related to a potency test in laboratory animals. The highest mean hemagglutination inhibition antibody response in horses occurred in groups vaccinated, respectively, with 128 or 256 hemagglutination units of A/Equi-1 and 512 or 1024 hemagglutination units of A/Equi-2 antigen. Groups vaccinated with further two- or fourfold increases in these antigens had mean hemagglutination inhibition titers that were somewhat lower than the maximum levels. When graded doses of vaccine were given to guinea pigs, their hemagglutination inhibition antibody titers reached a plateau of maximum values, similar to the serological response in vaccinated horses. Test horses remained clinically free from signs of equine influenza during the year following vaccination and no untoward post-vaccination reactions were observed.


Proximal-distal absorptive gradients in the in vivo intestine of normal and infected (Hymenolepis diminuta: Cestoda) rats. Using a single-pass perfusion technique, H2O, Na+, Cl-, HCO3-, and glucose absorption were studied in the jejunum and proximal and distal ileum of rats either uninfected or infected with a tapeworm parasite (Hymenolepis diminuta). The effect of parasitization, region of the intestine, type of buffer, and concentration of glucose in the perfusion fluids on the transport data were analyzed by univariate and multivariate techniques. Proximal-distal flux gradients were observed for water and all the solute species studied, as well as for glucose- and bicarbonate-stimulated salt and water transport; there was a decreasing sensitivity to low pH proceeding distally. The major regional differences occurred between the proximal and distal ileum, with the fluxes in the jejunum being similar to those in the proximal ileum. Na+, H2O, and glucose transport decreased, while Cl- absorption increased, proceeding distally. The parasites diminished the rates of absorption of glucose, salt, and water, and altered the flux gradients, particularly the Na+ and HCO3- transport gradients. The differences in the gradients between control and infected animals were related to differential sensitivity of the different transport systems in the various regions of the gut to parasitism.


Influence of cold exposure and thyroid hormones on regulation of adrenal catecholamines. Since thyroid hormones influence urinary excretion of catecholamines after exposure to cold, the effects of hyper- and hypo-thyroidism on adrenal tyrosine hydroxylase (TH) (EC 1.14.16.2), phenylethanolamine-N-methyl transferase (PNMT) (EC 2.1.1.28), and serum dopamine-beta-hydroxylase (DbetaH) (EC 1.14.17.1) of rats of 23 and 4 degrees C were studied. TH changes resembled the urinary excretion pattern at 4 degrees C in being higher after 8 days than after 1 day of exposure, and in declining as acclimation occurred. At 23 degrees C, TH activity of hypothyroid rats was significantly higher than in euthyroid or hyperthyroid animals, and after 1 day at 4 degrees C the value increased even more. While in the hypothyroid animals at 4 degrees C the concentration of adrenal catecholamines was less, the epinephrine to norepinephrine ratio was higher than at 23 degrees C. Very high TH activity with a decline in catecholamine concentration suggests that the capacity of TH had been exceeded. PNMT activity was significantly elevated in this group. TH activity was not decreased in the hyperthyroid group at 23 degrees C, and was increased after 8 days at 4 degrees C, suggesting that circulating thyroid hormones have no direct inhibitory effect on TH. Serum DbetaH was elevated after exposure to 4 degrees C, regardless of thyroid hormonal status. The activation of adrenal TH in hypothyroid rats at 23 degrees C and of TH, PNMT, and serum DbetaH at 4 degrees C is probably the result of increased activity of the sympathetic nervous system.


The catabolism of L-tyrosine by an Arthrobacter sp. An Arthrobacter sp. metabolizes L-tyrosine by a pathway involving 3,4-dihydroxyphenylacetate as a key intermediate. p-Hydroxyphenylpyruvate is formed from tyrosine by an amino-transferase specifically requiring alpha-ketoglutarate for activity, and is then converted to p-hydroxyphenylacetate by an oxidative decarboxylation. p-Hydroxyphenylacetaldehyde is not an intermediate in the formation of p-hydroxyphenylacetate. Extracts of the bacterium oxidize 3,4-dihydroxyphenylacetate to delta-carboxymethyl-alpha-hydroxymuconic acid which, when supplemented with 2 mol of diphosphopyridine dinucleotide, results in the production of stoichiometric amounts of succinate and pyruvate.


The cellulase system of a Cytophaga species. Cellulases (EC 3.2.1.4) of a Cytophaga species WTHC 2421 (ATCC 29474) were found in the soluble portion of the cell (the periplasm and the cytoplasm) and on the membrane. Cell-free cellulases were not found. Most of the carboxymethylcellulase activity associated with reduction of viscosity was membrane bound, whereas most of the carboxymethylcellulose (CMC) saccharifying activity was soluble. The CMC-saccharifying activity was increased 534 X by purification procedures which included ammonium sulfate precipitation and molecular exclusion chromatography with Sephadex G-75 and Biogel p-100. Periplasmic carboxymethycellulase had a molecular weight of 6250 and cytoplasmic carboxymethylcellulase had a molecular weight of 8650. Analytical ultracentrifugation of the periplasmic carboxymethylcellulase (CMCase) indicated that it had a low molecular density. The chromatographic fraction containing periplasmic CMCase also contained enzyme activity against crystalline cellulose. The activity against crystalline cellulose was 238 X higher than the activity shown by the whole cell. The reaction of the enzyme with either CMC or dewaxed cotton produced only glucose. The enzyme was slightly inhibited by the presence of 0.01% (w/v) glucose, lactose, or cellobiose, but it was not affected by sucrose, and exhibited increased activity in the presence of xylose and fructose.


The role of 6-phosphogluconate dehydrogenase in Rhizobium. A nicotinamide adenine dinucleotide (NAD) linked 6-phosphogluconate (6-PG)dehydrogenase has been detected in Rhizobium. The enzyme activity is similar in both slow- and fast-growing rhizobia. The nicotinamide adenine dinucleotide phosphate (NADP) dependent 6-PG dehydrogenase was detected only in the fast growers and was more than twice as active as the NAD-linked enzyme. Partial characterization of the products of 6-PG oxidation in Rhizobium suggests that the NADP-linked enzyme is the decarboxylating enzyme of the pentose phosphate (PP) pathway (EC 1.1.1.44) whereas a phosphorylated six-carbon compound, containing ketonic group(s), is the product of the oxidation catalyzed by the NAD-linked enzyme.


Glycogen synthesis by amylosucrase from Neisseria perflava. Amylosucrase (sucrose:1,4-alpha-D-glucan 4-alpha-glucosyltransferase; EC 2.4.1.4) which mediates the transfer of the glucosyl moiety of sucrose to a growing alpha-1,4-glucan chain is a constitutive enzyme of Neisseria perflava. The products of enzymic action are insoluble glycogenlike polysaccharides and fructose, the latter being a competitive inhibitor of the enzyme (Ki=20 mM). The enzyme is extremely stable and appears to bind very tightly to its polymerized product. Properties of product-bound enzyme reflect those of the native complex.


Further development of a successful protocol of graft versus leukemia without fatal graft-versus-host disease in AKR mice. We previously reported a successful model for treatment of BW 5147 leukemia in AKR mice by adoptive immunotherapy using allogeneic spleen cells from C57BL/6 mice. The leukemia cells were given 3 days before initiation of therapy. Graft-versus-host reaction was prevented by treatment with spleen cells from a second allogeneic strain (CBA), followed by cyclophosphamide and syngeneic spleen cells. We now show that it is not necessary to use syngeneic spleen cells in the final transplant since H-2-compatible, allogeneic CBA cells are as effective. In addition, it is possible to initiate successful therapy 5 days after leukemia implantation providing that the initial cyclophosphamide, given in two doses of 100 mg/kg each and spaced 7 days apart, is administered prior to establishment of graft-versus-host reaction. Higher single doses of drugs were followed by fatal graft-versus-host disease.


Glucocorticoid receptors in Morris hepatomas and host liver and the correlation of biological activity with receptor levels. Glucocorticoid-binding macromolecules were examined in Morris hepatomas 7787, 5123tc, 3683F, 7800, and 3683 and the Reuber hepatoma H-35 with the use of the synthetic glucocorticoid, triamcinolone acetonide. The physical properties of the triamcinolone acetonide-binding macromolecules of the hepatomas indicate that they are specific glucocorticoid receptors. The equilibrium association constants (Ka), sedimentation coefficients, and sensitivity to sulfhydryl-blocking reagents were found to be similar when hepatoma receptors were compared with the known properties of the liver receptor. Probably the most convincing criterion that the triamcinolone acetonide-binding macromolecules from the hepatomas are specific receptors is that 50 to 90% of the receptor can be depleted from hepatoma cytosol by treating rats with cortisol. In adrenalectomized tumor-bearing rats, the receptor levels in hepatomas 7787, 7800, 5123tc, and H-35 are comparable to or greater than receptor levels of host liver. However, tryptophan oxygenase was not responsive to glucocorticoids in hepatoma 7800 although receptor levels were quite high, and there were no indications that the receptor molecules were altered. Hepatomas 3683 and 3686F have low levels of receptor which may be related to resistance of these tumors to glucocorticoid treatment.


Production of calcitonin, adrenocorticotropic hormone, and beta-melanocyte-stimulating hormone in tumors derived from amine precursor uptake and decarboxylation cells. The tumor production of human calcitonin (CT) was examined by radioimmunoassay, and it was found that 50 of 85 (59%) tumor tissues collected at random contained immunoreactive CT. These tumors were grouped as to whether they were derived from the amine precursor uptake and decarboxylation (APUD) series. The group that was derived from APUD cells showed appreciable amounts of CT in 30 of 31 (97%) of these tumors or in 20 of 21 (95%) when the medullary carcinomas of the thyroid were excluded. However, of the non-APUD group of tumors only 20 of 54 (37%) were found to contain CT, so that the difference between these two groups was highly significant (p less than 0.001). Of the tumors with ectopic adrenocorticotropic hormone-melanocyte-stimulating hormone production, 12 of 14 were shown to contain CT. These data indicate that CT is a common product of the APUD tumors and that tumor production of CT is often associated with that of adrenocorticotropic hormone and beta-melanocyte-stimulating hormone.


The use of titanium (IV) oxide for the immobilisation of carbohydrate-directed enzymes. The use of particles of porous titanium (IV) oxide as a suitable matrix for enzyme immobilisation has been investigated with dextranase. Treatment of the particles with enzyme in the presence and absence of ammonium ions showed that the presence of ammonia induced a greater coupling of protein, whereas a greater retention of enzyme specific activity was achieved in the absence of ammonia. Properties of the immobilised enzyme include a pH-dependence and reversibility of the coupling between enzyme and matrix. The immobilised dextranase was most stable at pH 5.0. Automated analytical techniques for measuring the activity of dextranase and other polysaccharidases in soluble and insoluble forms are also reported.


[Regulation and electric excitation of enzyme membranes in vitro]. Enzyme membranes can be activated or inhibited by applying continuous or alternating electrical fields. The field can modify the transport or reaction term of the transport-reaction by action on the displacement of charged species including those giving pH effects or inducing volume flows. A first experimental example is given: the progressive supression of the inhibition of hexokinase by the product when increasing alternating fields are applied. In the same way the apparent optimal pH approaches that of the soluble enzyme. In addition to its theoretical and practical implications electrical regulation can lead to the monitoring of enzyme reaction-driven mechanochemical fibers.


A kinetic spectrophotometric assay for rapid determination of acetoacetate in blood. We describe an automated kinetic assay for acetoacetate in blood. Acetoacetate is enzymatically reduced to D-beta-hydroxybutyrate and the reaction is monitored for 60 s with a reaction-rate analyzer. This technique allows low concentrations of acetoacetate to be measured with good precision and overcomes many of the problems associated with other automated techniques. Our studies on the stability of acetoacetate emphasize the need for care in handling specimens. The use of a reaction-rate analyzer, an item of equipment common to most laboratories, allows for rapid handling of samples in small or large batches, depending on the needs of the laboratory.


Automated kinetic determination of lactate dehydrogenase isoenzymes in serum. A steady-state kinetic method has been revised for measuring lactate dehydrogenase isoenzyme activities, which relates the inhibition of heart-type isoenzyme activity to the overall isoenzyme composition of the enzyme subunits. The method depends on the pH-dependent formation of an inhibitory ternary complex by the heart-type isoenzyme with NAD+ and pyruvate (if the reaction is measured by NADH oxidation). A preincubation step in the previous method is eliminated. The isoenzymes are measured by measuring the reduction of pyruvate in two different concentrations, which favor either the total or fractional activity, depending on the concentrations of pyruvate and the percentage of heart-type subunits. The method has been adapted to a centrifugal analyzer, which has speeded automated isoenzyme determinations, with an accuracy comparable to that for electrophoretic methods.


Determination of free and esterified cholesterol by a kinetic method. I. The introduction of the enzymatic method with 2,2'-azino-di-3[ethyl-benzthiazolin sulfonic acid (6)] (ABTS). A new, simple kinetic method is described for the determination of serum cholesterol based on the oxidation of 2,2'-azino-di-3[ethyl-benzthiazolin sulfonic acid (6)] (ABTS) by use of a single aqueous reagent. This assay procedure is rapid, specific, reproducible and applicable to the measurement of free and esterified cholesterol in a continuous procedure. The method requires no prior treatment of sample and linear kinetics are obtained up to 13 mmol/l cholesterol. The use of ABTS permits the direct calculation of cholesterol concentrations from absorbance changes at 410 nm (A = epsilon - c - d).


[Concanavalin A- Sepharose affinity chromatography for routine microanalysis of gamma-glutamyltransferase (author's transl)]. The separation of two molecular forms of liver gamma-glutamyltransferase is achieved by Con A-Sepharose chromatography, an adult type with high affinity to Con A and a fetal type without binding capacity to this lectin. Now we present a new method using micro-columns for affinity chromatography (gel volume 2 ml). By this rapid and inexpensive procedure it is possible to study this enzyme separation for its use in the diagnosis of liver diseases.


Determination of serum acid phosphatase in Gaucher's disease using 4-methylumbelliferyl phosphate. We describe a new assay that is useful for identifying individuals who may be affected with Gaucher's disease. The assay involves the determination of serum acid phosphatase activity using the fluorogenic substrate 4-methylumbelliferyl phosphate. The assay measures acid phosphatase activity at pH 6.0 in the presence of 3.0 M 2-mercaptoethanol and requires a 5 microliter serum sample and a 15-min incubation period. Under these conditions, 2-mercaptoethanol preferentially inhibited the acid phosphatase activity in control serum but did not inhibit the elevated acid phosphatase present in the serum of patients with Gaucher's disease. Using this assay, we observed a 5-50-fold elevation in serum acid phosphatase activity in 8 patients with the adult, non-neuropathic form of Gaucher's disease when compared to control serum assayed under the same conditions. Serum from several heterozygotes free from pathology exhibited normal acid phosphatase activity when assayed at pH 6.0 in the presence of 2-mercaptoethanol. Acid phosphatase activity in serum from patients with prostatic cancer can be distinguished from that in Gaucher serum on the basis of the well-documented sensitivity of the former to inhibition by sodium tartrate. A serum sample from a patient with Niemann-Pick disease exhibited a mild elevation in tartrate-resistant acid phosphatase activity so that conclusive diagnosis of Gaucher's disease requires assaying leukocytes or fibroblasts from suspected patients for glucocerebroside:beta-glucosidase activity.


New enzymatic assay of cholinesterase activity. A new enzymatic method for the determination of serum pseudo-cholinesterase activity is described. Choline, which is liberated from benzoylcholine as substrate by cholinesterase, is oxidized by choline oxidase to betaine with the simultaneous production of hydrogen peroxide, which oxidatively couples with 4-aminoantipyrine and phenol in the presence of peroxidase to yield a chromogen with maximal absorbance at 500 nm. The calibration curve is linear up to 1500 units per liter of serum. The method is reproducible, and the results correlate well with those obtained by the method using butyrylthiocholine as substrate and 5,5'-dithiobis-(2-nitrobenzoic acid) as color reagent.


Quantitative elution studies in experimental immune complex and nephrotoxic nephritis. Optimal conditions have been determined for elution of antiglomerular basement-membrane antibodies and antigen-antibody complexes from nephritic kidneys, obtained from rabbits with nephrotoxic nephritis and various forms of BSA-induced immune complex disease. Elution from kidney homogenates was performed with acid and alkaline buffers, solutions of chaotropes, urea, sodium and magnesium chloride and other agents. Functional activity of antibodies exposed to elution procedures was tested by immunofluorescence, or by a modified Farr's technique. Antibodies of progressively greater binding capacity were eluted by a stepwise acid gradient (pH 3-2--2-8) using low or high ionic strength glycine--HCl buffer. Antigen-antibody complexes were best eluted using 3 M KBr (pH 9) using several extractive steps. A stepwise alkaline gradient (pH 10-5--11-1 using 0-1 M glycine-NaOH buffer) or 8 M urea were found to be satisfactory alternative methods of eluting immune complexes. Elution procedures were best carried out in the cold in all circumstances. Other eluting agents were found to be less successful or less practical.


Cyclic 3'5'-adenosine monophosphate and central circulatory control in cats and dogs. 1. Dibutyryl cyclic AMP (Db cAMP, 75-500 microgram/kg), injected into the lateral ventricle of the brain of the cat increased blood pressure, heart rate and splanchnic discharge rate. 2. ATP, but not AMP, induced similar changes; GMP in small doses increased blood pressure. 3. A number of drugs are known to activate adenylate cyclase-induced hypertension, tachycardia and increase splanchnic discharge rate. This was shown for TRH, tetracosactide and a new beta2-adrenoceptor stimulant, NAB 365. 4. Injection into the lateral ventricle of theophylline or Ro 7/2956, both inhibitors of phosphodiesterase, similarly increased blood pressure. 5. Histamine administered by the same route induced similar reactions; it is not known if this action was exerted by activation of H1- or H2-receptors. 6. Somatostatin, known to reduce cAMP levels, induced a small but significant decrease in blood pressure. Melanocyte stimulating hormone release inhibiting factor (MIF) and TSH were ineffective. 7. These results provide evidence for the possibility of a role for cAMP in the central regulation of blood pressure at suprabulbar levels.


Effects of urine acidification on plasma and urine phencyclidine levels in overdosage. A gas chromatography--mass fragmentography--electron impact (GC-MS-EI) assay of phencyclidine (PCP) was adapted for human plasma and urine. This assay is specific for PCP and very sensitive (approximately 1 ng/ml). Patients with the putative diagnosis of PCP overdosage were studied to correlate plasma and urinary levels with clinical state. Urinary PCP levels were enhanced in an acid urine, which suggests that acidification of the urine is an adjunct in the therapy of PCP overdosage.


[Modified agar diffusion test for direct testing of the effectiveness of released antibiotics from bone cements]. A modified agar-diffusion method is used to test the bactericidal efficacy of antibacterial substances (model substance: Gentamicin) that have been mixed with Palacos osseous cement. Holes of a diameter slightly larger than the diameter of the cylindric Palacos test block are cut from the agar plate. The remaining space (inner wall of the hole--edge of the test body) is filled with various eluate media. The release of active antibacterial substances from polymethylmetacrylates can thereby be determined under nearly physiologic conditions at varying pHs by direct antibacterial testing.


Clorazepate: double blind crossover comparison of a single nightly dose with diazepam thrice daily in anxiety. In a double blind cross-over study in 40 patients with mild to moderate anxiety, clorazepate 15 mg at bedtime was as effective as diazepam on global rating and slightly superior on target symptom assessment. There was a significantly higher incidence and frequency of side effects during diazepam treatment. Occurrence of side effect related to plasma diazepam, and anxiolytic effect related to plasma nordiazepam. These results are discussed in relation to convenience of the single dose regimen and psychomotor performance.


Effect of dose on the pharmacokinetics of intravenous nicotine in the rat. The pharmacokinetics of [methyl-14C]nicotine were evaluated in male Fischer-344 rats following the iv injection of 0.08-, 0.4-, or 0.8-mg/kg doses. Independent of dose, the concentration of nicotine declined biexponentially with a mean half-life ranging from 0.92 to 1.10 hr. However, the total plasma clearance of nicotine increased (p less than 0.05) from a mean value of 2.53 liter/hr-kg at the 0.08 mg/kg dose to mean values of 3.88 and 4.04 at the 0.4- and 0.8-mg/kg doses, respectively. There were no dose-related differences in the half-life of formation of the major metabolite, cotinine (range 0.33-0.46 hr) or the half-life of its elimination (range 5.39-6.63 hr). However, the areas under the cotinine plasma concentration-time curves did not increase proportionately with the dose of nicotine, indicating that the dose-related increase in the total plasma clearance of nicotine is not due to an increase in its biotransformation to cotinine. Similar conclusions were obtained with regard to the disposition kinetics of other polar metabolites of nicotine. The urinary excretions of nicotine and its metabolites were dose-independent.


Pharmacokinetics of 3,3',5,5'-tetrachlorobiphenyl in the male rat. The pharmacokinetics of 3,3',5,5'-tetrachlorobiphenyl (4-CB) was studied to determine the importance of chlorine position to the kinetics of polychlorinated biphenyl (PCB) distribution, metabolism, and excretion by the rat. Tissue samples and excreta were collected at times ranging from 2 hr to 42 days after intravenous injection of a single 0.6-mg/kg dose of 14C-labeled 4-CB. The relative fractions of 4-CB and its metabolites were determined in selected samples, and 4-hr bile cannulations were performed to establish the rate of biliary excretion. A mathematical model description of PCB kinetics in the rat was defined here for 4-CB and used to generate tissue and excretion concentration-time profiles for both the parent compound and its metabolites. Model predictions were in good agreement with the experimental data, and showed how the individual pharmacokinetics of 4-CB compared to several other PBC's. The rate of 4-CB metabolism and excretion was slower than would be predicted according to its degree of chlorination. The relative rates of metabolism for 4-CB and five other differently chlorinated biphenyls could be ordered according to the number of adjacent unsubstituted carbon-atom pairs they contained. It appears that chlorine position is more important than degree of chlorination because the former can alter the availability of vicinal hydrogens thought to facilitate rapid PCB metabolism by mammals.


Alteration of in vivo zoxazolamine metabolism by carbon monoxide in normal and polycyclic hydrocarbon-treated immature male rats. The effect of carbon monoxide exposure on the in vivo metabolism  of zoxazolamine in normal (corn oil-treated) and 3,4-benzpyrene- and 3-methylcholanthrene-treated, immature, male rats was examined. Pre-exposure of the animals for 90 min followed by determination of the duration of drug action while the animals were maintained in the experimental atmosphere resulted in a qualitative difference in response between normal and polycyclic hydrocarbon-treated animals over the concentration range of 150-450 ppm CO. Corn oil-treated animals demonstrated a decreased duration of drug action when exposed to CO, indicating an increase in the bioavailability of drug for metabolism which may be a result of an increase in liver perfusion rates. In contrast, polycyclic hydrocarbon-treated animals demonstrated an increased duration of drug action on exposure to CO. The qualitative difference in response to CO exposure in the two groups of animals may be due to differences in the sensitivity of cytochrome P-450 and cytochrome P-448 with respect to complexing with CO or to lowered intracellular PO2; or to differences in the dependency on blood flow of the rate of in vivo metabolism of zoxazolamine.


Developmental aspects of xenobiotic transformation. In most laboratory animals monooxygenases are apparently absent or barely detectable in fetal organs until just before birth. In this contribution hepatic cytochrome P-450-dependent reactions in the rat are considered only. The results are interpreted on basis of the reaction scheme of Estabrook. To avoid methodological pitfalls the basic kinetics for all reactions investigated have been investigated with liver preparations from newborn and adult rats. The low monooxygenase activity of rat liver during the perinatal period can be observed even under optimal conditions for the in vitro enzyme assay. There are different developmental patterns for various reactions O-demethylation of codeine, phenazone-hydroxylation, first and second steps on N-demethylation of amidopyrine, N-demethylation of ethylmorphine. There are marked differences not only in Vmax but also in the postnatal development of Km and the inductibility by phenobarbital. Thus the existence of a different cytochrome P-450 is evident also by this approach. The low monooxygenase activity of rat liver during the perinatal period is not due to a lack of NADPH or NADH, to an age-dependent NADPH cytochrome P-450 reductase activity or to an age-dependent NADH-cytochrome P-450 reduction. Moreover this low activity is not due to an insufficient mitochondria-endoplasmic reticulum interaction. It is accompanied by low delta Amax after addition of a typical type I substrate (hexobarbital) and by a small amount of metyrapone-binding centers: it can be explained by a smaller percentage of active cytochrome P-450 in comparison to adult rat liver.


A kinetic study of the pH-dependent properties of the ferric undecapeptide of cytochrome c (microperoxidase). The ferric form of the haem undecapeptide, derived from horse cytochrome c by peptic digestion, undergoes at least three pH-induced transitions with pK values of 3.4, 5.8 and 7.6. Temperature-jump experiments suggest that the first of these is due to the binding of a deprotonated imidazole group to the feric iron while the second and third arise from the binding of the two available amino groups present (the alpha-NH2 of valine and the epsilon-NH2 of lysine). Molecular models indicate that steric retraints on the peptide dictate that these amino groups may only coordinate to iron atoms via intermolecular bonds, thus leading to the polymerization of the peptide. Cyanide binding studies are in agreement with these conclusions and also yield a value of 3.6 X 10(6) M-1 s-1 for the intrinsic combination constant of CN- anion with the haem. A model is proposed which describes the pH-dependent properties of the ferric undecapeptide.


The role of cytochrome P-450 in the hydroperoxide-catalyzed oxidation of alcohols by rat-liver microsomes. The organic hydroperoxide cumene hydroperoxide is capable of oxidizing ethanol to acetaldehyde in the presence of either catalase, purified cytochrome P-450 or rat liver microsomes. Other hemoproteins like horseradish peroxidase, cytochrome c or hemoglobin were ineffective. In addition to ethanol, higher alcohols like 1-propanol, 1-butanol and 1-pentanol are also oxidized to their corresponding aldehydes to a lesser extent. Other organic hydroxyperoxides will replace cumene hydroperoxide in oxidizing ethanol but less effectively. The cumene-hydroperoxide-dependent ethanol oxidation in microsomes was inhibited partially by cytochrome P-450 inhibitors but was unaffected by catalase inhibitors. Phenobarbital pretreatment of rats increased the specific activity of the cumene-hydroperoxide-dependent ethanol oxidation per mg of microsomes about seven-fold. The evidence suggests that cytochrome P-450 rather than catalase is the enzyme responsible for hydroperoxide-dependent ethanol oxidation. However, when H2O2 is used in place of cumene hydroperoxide, the microsomal ethanol oxidation closely resembles the catalase system.


Kinetics of sheep-liver cytoplasmic aldehyde dehydrogenase. 1. Sheep liver cytoplasmic aldehyde dehydrogenase showed little pH dependence of V or kcat. Some buffer anion effects were noted. 2. The oxidation of aldehydes at pH 7.6 was quantitative but irreversible. The initial velocity data indicated a sequential mechanism for the addition of substrates. Inhibition by NADH and the product analogue 2-bromo-2-phenylacetic acid, together with the known tight binding of NADH to the free enzyme, indicated an ordered mechanism with NAD+ as leading substrate. 3. Values for the rate of binding and dissociation of NAD+ were obtained from the steady-state data. The values obtained were virtually identical with those which could be calculated from the data for the horse liver cytoplasmic enzyme. Close similarities are in general apparent for the horse and sheep liver cytoplasmic enzymes and with other tissue aldehyde dehydrogenases. 4. Apparent substrate activation was observed with high concentrations of both acetaldehyde and propionaldehyde, a limiting value of 0.25s-1 being obtained for kcat. No isotope effect was observed on V using [1-2H]propionaldehyde as substrate suggesting that NADH release might be rate-limiting in the steady-state. 5. The implications of the non-linear steady-state behaviour are discussed.


Maldescent of the testes--an epidemiological study. The case histories of 2,362 newborn boys were evaluated for congenital testicular maldescent. These data, obtained from the prospective follow-up study "Pregnancy and Child Development" sponsored by the Deutsche Forschungsgemeinschaft, are discussed in the light of new findings on morphological changes in this disorder. The aetiology of maldescent of testes is considered to be heterogenous and could not be clarified by investigating 26 factors which might be regarded as possible causes of the abnormality. The difficulties in achieving a correct classification of the various forms of maldescended testes are emphasized and are demonstrated by data from this collaborative study. Considering the frequency of non-descent at the ages of 9 months (3.3%), 18 months (2.9%), and 36 months (2.5%), we believe we have found some evidence for spontaneous descent in a proportion of cases during the first 3 years of life. If this is confirmed by proposed new studies, the findings will have to be weighed against the recommendations of the 1974 International Health Foundation that therapy should be performed during the 2nd year of life or earlier.


Adult height of patients treated in childhood for undescended testes. 101 patients successfully treated for undescended testes with human chorionic gonadotrophin (HCG) or with HCG followed by surgery reached normal adult heights. Deviations from normal growth in individual patients did not appear to be related either to their earlier undescended testes or to the successful therapy, but rather to their genetic background. Therefore, apart from patients with chromosomal abnormalities and other primary causes for maldescent of the testes, boys with undescended testes show a normal distribution for height and do not represent a different population with regard to growth.


Adult metachromatic leukodystrophy. I. Clinical manifestation in a female aged 44 years, previously diagnosed in the preclinical state. In a 5-year follow-up of a case of adult metachromatic leukodystrophy, already diagnosed in the preclinical stage, the development of the symptoms of this disease could be studied in detail: initially, lack of drive, emotional lability and depressive mood. At the same time, pain in the arms and beginning gait disturbance. Later, impairment of memory and concentration, disorientation, inadequate behavior and progression of gait disturbance. Finally spastic atactic gait with small steps and dyspractic components, coordination disturbances with writing dysfunction, fast dysarthric speech, hyperkinetic activity, compulsory emotional outbursts and progressive dementia. Only minor neurological signs such as reflex abnormalities. In the EEG, slight slowing of frequencies compared to earlier tracings. Increasing diminution of nerve conduction velocity in the lower limbs. Only minor increase of CSF protein (51 mg%). In spite of normal vision, evoked visual potentials abnormal, response of optical and electrical blink reflexes delayed. Imperfect filling of gallbladder. No significant quantitative changes of the biochemical parameters compared with the findings made 5 years earlier (excretion of urinary sulfatides, diminished activity of arylfulfatase A in urine and leukocytes).


Adult metachromatic leukodystrophy. II. Ultrastructural findings in peripheral nerve and skeletal muscle. Sural nerve biopsy in a 44-year-old woman with adult metachromatic leukodystrophy (MLD) confirmed by deficient arylsulfatase-A activity, showed a reduction in the number of large and small myelinated axons, and sparse metachromatic material. Ultrastructurally, the latter consisted of various types of residual bodies including the tufaceous and prismatic forms typical of MLD. In the striated muscle, large amounts of regular lipofuscin but no MLD-characteristic inclusions were encountered. Inclusion-bearing mitochondria in the muscle appeared to be an incidental finding.


Carbohydrase activity in the digestive system of some teleost fishes. The activity of carbohydrases in Puntius sophore (Ham.), Channa gachua (Ham.) and Cirrhinus mrigala (Ham.) has been studied. The carbohydrases have been found in the stomach, intestinal bulb, intestine, pyloric caeca and the hepato-pancreas. The hepatopancreas is the main site of production of these enzymes and it is in this organ and the intestine that their activity is highest. Their pH optimum lies between 5.4 and 6.4. The enzyme equipment in the teleost is adapted to their respective food and feeding habits both qualitatively and quantitatively. In Puntius (omnivorous) and Cirrhinus (herbivorous) all three carbohydrases, namely amylase (EC 3.2.1.1.), sucrase (EC 3.2.1.26.) and raffinase, while in Channa (carnivorous) only amylase and sucrase have been found to be active. In Cirrhinus mrigala, which is predominantly a herbivorous species, the concentration of carbohydrases is higher than those in the other two fishes.


Aminotripeptidase (EC E.4.1.3) activity in the digestive system of some teleost fishes. Aminotripeptidase (EC 3.4.1.3) has been found to be present in the intestinal mucosa, hepatopancreas and the pyloric caeca of the three fished examined. The pH optimum of the enzyme ranges from 6.6 to 7.6. The pH optimum varies from species to species and also from section to section of the gut in the same species. The enzyme is most active in the carnivorous Channa gachua in comparison to the other fishes studied.


The interaction of 1-(2(diarylmethoxy)ethyl)aziridines with histamine receptors in the longitudinal muscle strip of the guinea pig ileum. The time course of the onset and decline of histamine antagonism by 1-(2-(diarylmethoxy)ethyl)aziridines, compounds which could be expected to have H1-receptor alkylating properties, was investigated on the longitudinal muscle strip of the guinea pig ileum. Experiments were performed with normal preparations and with muscle strips pretreated with a prostaglandin synthesis inhibitor in order to prevent spontaneous rise of muscle tone. In contrast to the previously reported observation that antihistaminic potency decreased with prolongation of the preincubation time, histamine antagonism by the aziridine compounds remained at a constant level for more than 60 min in the presence of indomethacin. This indicated that the aziridines are not hydrolyzed either directly in solution or after interaction with the tissue since the supposed hydrolysis product had a significantly lower antihistaminic activity. A comparison between 1-(2-diphenylmethoxy)ethyl)aziridine and diphenhydramine showed the former compound to have a slightly more rapid onset and a considerably more rapid decline of histamine receptor blockade. It was concluded that 1-(2-(diarylmethoxy)ethyl)aziridines did not alkylate the histamine H1-receptor in the longitudinal muscle layer of the guinea pig ileum.


[Comparative biochemical studies of placental alkaline phosphatase in animals (author's transl)]. Biochemical properties of alkaline phosphatase (ALP) from placenta were compared between man, dog, cat, rabbit and cattle. 1) Optimum pH of the enzyme was essentially identical through the species of the animals but the inhibition of L-phenylalanine was clearly demonstrable with human ALP but little with that of other animals. 2) ALP of human placenta was not inactivated by heating at 65 degrees C for 15 min. but one of the other animals was thermolabile. Such thermostability of human placental ALP almost disappeared after treatment with EDTA, suggesting that the divalent metal ions were required for the thermostability. 3) Activities of placental ALP were inhibited by cationsurfactants in human and rat but not in the other animals, while the inhibition by DOC-Na, an anion-surfactant, was seen only in human. 4) The affinity to beta-glycerophosphae of placental ALP was seen only in human.


Experimental renal papillary necrosis. Effects of beta adrenergic receptor blockade, heparin and hydrocortisone. Renal papillary necrosis was induced in rats by daily subcutaneous injection of 15 mg 2-bromoethylamine hydrobromide (BEA) per 100 g of body weight for 2 successive days. This dose was 50% higher than that reported previously. Beta adrenergic receptor blockade with oxyprenolol did not influence the kidney damage. The administration of heparin did not show any effect. The doses applied did not induce the incoagulability for a sufficient period of time. On the contrary, treatment with hydrocortisone decreased papillary necrosis without inducing increased diuresis.


Hypoxia in fibroblast cultures. 3. 35S-sulfate incorporation into acid mucopolysaccharides as influenced by 5% O2 hypoxia with simultaneous changes in the lactate-and H-ion concentrations. The influence of 5% O2 hypoxia on the 35S-sulfate incorporation into different mucopolysaccharide fractions was studied in monolayer cultures of embryonic rat fibroblasts before reaching the stationary phase. Besides the O2 concentration also the pH value and lactate concentration were varied in the experiments. The results are valid for proliferating cell cultures.


Hypoxia in fibroblast cultures. 4. Cell density, glucose utilization and acid mucopolysaccharides in 1% O2 concentration. Secondary cultures of embryonic rat fibroblasts incubated in Eagle medium with Hepes buffer at pH 6.6 and 7.4 were maintained in hypoxia of 1% O2 for 48 hours. The acid mucopolysaccharides were isolated from medium plus cells and fractionated on cellulose microcolumns according to SVEJCAR and ROBERTSON (1967). Glucose utilization, lactate production, quantity of total MPS and changes in the MPS distribution pattern were correlated with cell density.


The problem of sea urchin egg fertilization and its implications for biological studies. The analysis of sea urchin egg fertilization shows that several phenomena common to other biological systems are involved: cell recognition, cell fusion, exocytosis and initiation of mitotic activity. Both the role of calcium ions in cell fusion and exocytosis and the function of the cell surface in the initiation of mitotic activity appear to have general applicability. The study of fertilization can contribute to the elucidation of these processes and, reciprocally, progress in this field can help to advance our understanding of the mechanisms of fertilization in sea urchins and other organisms.


Hydrogen peroxide generation in Trypanosoma cruzi. Homogenates from T. cruzi epimastigotes produced 3.4 pmoles H2O2/min 10(6) cells, as detected by the cytochrome c peroxidase assay. Addition of NADH or NADPH increased H2O2 production by a factor of 3 and 5, respectively. When supplemented with NADH and NADPH, the mitochondrial, microsomal and supernatant fractions produced H2O2, the soluble fraction and the mitochondrial membranes being apparently the main generators of H2O2. The epimastigote homogenates showed cyanide-sensitive superoxide dismutase activity, equivalent to 0.28 microgram bovine superoxide dismutase per mg homogenate protein.


[Effect of several neuroleptic, adreno-, sympatho- and cholinolytic substances on the development of experimental cerebral edema induced by nicotine]. The influence of some neuroleptic, adreno, sympatho- and cholinolytic substances on the development of experimental brain edema induced with nicotine was studied in tests conducted on rats. It was ascertained that marked antiedemic properties display drugs blocking the alpha-adrenoreceptors (phentolamine, dopegit), the neuroleptic chlorpromazine and central M-cholinolytics (benactizine). Weak action exert central H-cholinolytics (difacil trasentin) and the neuroleptic galoperidol. Sympatho- and beta-adrenolytics (guanethidine, alpha-methyltyrosine, obsidan), and also the neuroleptic triphthazine fail to prevent the development of an experimental brain edema. It is presumed that a brain edema induced with nicotine comes as a result of the catecholamines liberation and is also due to stimulation of alpha-adrenoreactive systems.


[Pharmacologic study of clofelin].

[Pharmacologic study of clofelin]. Clofelin--2-(2,6-dichlorphenylamino)-2-imidasoline, an analoque of foreign-made clonidine (St155, catapresan, hemiton, etc) was subjected to a pharmacological study. In animals the drug produces first a rise and then protracted fall of the arterial pressure, reduces the frequency of cardiac contractions, lessens reflex hypertension that develops on compression of carotid arteries. When used in low doses it depresses the motor activity in unanesthetized animals and produces symptoms that bear evidence to its adrenomimetic action, e.g. exophthalmos, piloerection. Hypertension provoked by clofelin is caused by its peripheral and hypotension--by the central alpha-adrenomimetic action. The results of the present investigation prove clofelin to be identical with corresponding foreign drugs.


[Mechanism of the antianginal action of oxyfedrine]. Oxyphedrine was found to depress the tonicity of coronary vessels, increase the coronary circulation volume, the oxygen uptake by the heart and also to exert a positive inoand chronotropic action in test with urethan and cloralose anesthetized cats. These effects were caused by stimulation of the beta-adrenoreceptors of the heart and vessels. In non-anesthetized cats oxyphedrine did not produce tachycardia. Furthermore, under certain conditions (when superimposed on a preliminary administration of practolol- a beta-blocking agent) oxyphedrine can exert a direct inhibitory action on the myocardium.


Peinamine, a new bisbenzylisoquinoline alkaloid from arrow tips (pei-namô) of the Upper Orinoco. A new bisbenzylisoquinoline alkaloid has been isolated from a curare of the Upper Orinoco region. The curare is made by Indians from an unknown plant (probably Menispermacea) and is kept on arrow tips. The structure of the alkaloid, named peinamine, has been elucidated on the basis of its chemical reactions and spectroscopic data (N.M.R., M.S. and O.R.D.).


[Study of phenothiazine binding to membranes by the fluorescent probe method]. Binding of chlorpromazine, etaperazine and triftazine with model phospholipid membranes was investigated by using a 3-methoxybenzanthrone (MBA) fluorescent probe. All these preparations interacted with the membranes, this being registered by the MBA fluorescence extinction. The phenothiazine molecules bound with the membrane are localized in its superficial layer and extinguish the MBA fluorescence in different ways. The chlorpromazine binding was heterogenous and there are, apparently, 2 different ways of its molecules combination with the membranes. The effective extinction of MBA by phenothiazine increased in the following order: chlorpromazine, etaperazine, triftazine.


Effect of dibenzepin hydrochloride on the oxygen, glucose uptake and oxidative phosphorylation of rat brain in vitro. Dibenzepin hydrochloride (10(-3) M) decreases the oxygen uptake of whole brain homogenates and uncouples brain mitochondrial oxidative phosphorylation. At the same concentration this drug inhibits oxygen uptake of rat brain slices when calcium is absent in the incubation or then contains an excess of potassium.


[Long-term experiments on toleration of the ganglionic blockader hygronium]. In tests set up on albino rats, guinea pigs and rabbits the chronic toxicity of hygronium--a new original ganglion blocking agent of a short-term action was studied. With its daily intraperitoneal administration for 14 days in doses of 10 and 40 mg/kg (1/10 and 1/3 DL50) the drug produces in some of the animals, chiefly in rabbits and guinea pigs receiving 40 mg/kg, readily reversible histological changes in some of their internal organs. Hygronium does not cause any local irritating effect


[Effect of contrast media on the rate of oxidation of NADP.H and NAD.H by rat liver microsomes]. The effect of roentgen-contrast media on the activity of the NADP'N and NAD'N-dependent electron-transport chains of the rats liver microsomes was studied. Bilignost, cardiotrast, triiotrast and triombrin are shown to lower the rate of the NADP'N oxidation by the rats' liver microsomes and have no effect (except for triiotrast) upon the rate of the NAD'N oxidation. It is presumed that the relative resistance of the NAD'N-specific flavoproteid is due to the presence of a hydrophobic layer impervious to the polar molecules of the contrast media.


Excretion in insects: function of gut and rectum in concentrating and diluting the urine. The diverse excretory systems of insects exhibit several features that appear unusual when comparisons are made with the mammalian kidney. Secretion by the Malpighian tubules of a fluid that is unlike the blood in composition, substitutes for glomerular filtration. Various reabsorptive functions, such as volume reduction, regulation of individual electrolytes, adjustment of osmotic concentration and pH regulation, which are associated with distinct renal segments in the mammalian kidney, all occur simultaneously in the rectum of terrestrial insects. Involvement of an extracellular molecular sieve in selective reabsorption is novel. As far as water transport is concerned, the rectal pads of the cockroach and locust appear to accomplish, across a single layer of cells, the same function as the countercurrent multiplier system of the mammalian kidney with its several epithelial layers. Direct absorption of water vapor in the rectum of some insects from atmospheres of low relative humidity, clearly involves quite different and unknown mechanisms. Finally, saline-water insect larvae produce hyperosmotic excreta by direct secretion of ions into the rectal lumen. They can adjust individual transport processes to form various secretions, which are appropriate to the natural waters of diverse chemical types in which these larvae thrive.


The frequency of Y chromatin-positive spermatozoa during in vitro penetration tests under various experimental conditions. Complete analysis were carried out on the semen of 55 subjects. The semen was then investigated by using the fluorescent microscopic technique to determine the percentage proportion of Y chromatin-bearing spermatozoa. After staining with quinacrine mustard, an average Y chromatin frequency of 48.2% +/- 2.9% was recorded. With this spermatozoal material, 139 in vitro penetration tests were carried out with cervical mucus. After penetration there was a small, but statistically significant, increase in the percentage of Y spermatozoa (to 52.7% +/- 3.8%). The figures thus recorded were then correlated under variable experimental conditions in addition to variable semen parameters. The ratio of X:Y spermatozoa was related to the duration of the penetration tests, varying temperatures during the penetration, varying acid values of semen and cervical secretion, and also to the value of sperm motility in the semen analysis and to cervical factors. None of the above variables had any measurable influence on the percentage distribution of X and Y spermatozoa during in vitro penetration.


Xenogeneic neoplastic transplants in adult rats pretreated with thymic foetal extracts. The paper deals with the effect of thymic extracts from young foetal pigs and calves on the development of tumour xenografts, i.e. mouse ascites Ehrlich carcinoma and hamster malignant melanoma, in rats. The results of experiments showed that Ehrlich carcinoma developed in 4 out of 72 animals, i.e., in those animals which had received thymic extracts from young foetal pigs, Ehrlich carcinoma led to their death. Ehrlich carcinoma assumed a solid form and the cells changed in their shape. It developed normally, when retransplanted to mice. Hamster melanoma, when transplanted to rats given thymic extracts from young foetal calves, developed and survived for long periods of time (in one animal up to 190 days, and in the remaining animals much longer than in the controls and the animals injected with liver extracts from young foetal calves). The melanoma xenograft was not morphologically changed. A normal development of the malignant process occurred when melanoma was retransplanted to hamsters. Animals injected with thymic extracts from young foetal calves exhibited lymphopoenia and a fall in gamma globulin levels before tumour transplantation.


Influence of steroid structure in relation to liver metabolism during endotoxin lethality in mice. The influence of a number of steroid molecules on hepatic metabolism was determined in relationship to their ability to alter endotoxin lethality in intact mice. Progesterone, testosterone, estradiol, pregnenolone-16-alpha-carbonitrile and spironolactone did not alter endotoxin lethality, liver glycogen or tryptophan pyrrolase (TP) levels; all these materials increased liver tyrosine transaminase (TT) levels most probably due to mediation by endogenously liberated glucocorticoid hormones. Aldosterone and deoxycorticosterone induced liver TP, TT and glycogen but did not protect against lethality. Cortisone, hydrocortisone, triamcinolone, dexamethasone and 9-alpha-fluorohydrocortisone protected against lethality, increased liver glycogen and TT, but only triamcinolone induced liver TP. Collectively, there was no clear relationship between the protective effect and the increase or decrease of a given liver process. These further emphasize the need to reconsider molecular mechanisms of endotoxic reactions.


A comparison of D & C and vacuum aspiration for performing first trimester abortion. In a study of techniques for first trimester abortion conducted at Kandang Kerbau Hospital in Singapore from September 1973 to April 1975, 420 physically healthy gravidas were randomly assigned to one of two treatment groups either by D & C or vacuum aspiration. Half of the patients in each group were 6 to 10 menstrual weeks' gestation and half were 11 to 12 week's gestation. The numbers of women with one or more complications were the same for the two treatment groups, but were significantly less when termination was carried out at 11 to 12 weeks' gestation. Estimated blood loss was significantly lower for patients treated by VA (32.3 ml) than for patients treated by D & C (39.0 ml) at 6 to 10 week's gestation, but the difference in blood loss was not statistically significant at 11 to 12 weeks' gestation. Time from insertion to removal of the speculum was significantly less with VA than D & C at 6 to 10 weeks' gestation but not at 11 to 12 weeks' gestation. Since vacuum aspiration is at least as safe as D & C, and appears to be more convenient and costeffective, it should on the basis of this experience be accepted over D & C as the standard method for treatment of first trimester abortion.


Female sterilization using the tubal ring. The surgical and early postoperative complications were evaluated in a study of 221 laparoscopies and 29 minilaparotomies in which tubal occlusions were performed with the application of tubal rings. For either procedure, complications were infrequent. None of the procedures had to be completed by another technique of tubal occlusion, and none of the patients required an extended hospitalization for the treatment of a complication. Among the 79 patients who have been followed up for more than 6 months, no pregnancies have been reported. The tubal ring technique appears to be safe and effective when used with either laparoscopy of minilaparotomy.


Contraceptive efficacy of two different metals using a modified seven vector. Early studies with copper suggested that few improvements in overall IUD performance could be obtained with the use of more than 200 mm2 of copper. Thus, the use of other metals paired with copper has been investigated. This report of two studies of the small Copper-7, one with zinc and the other with copper alone, suggests that the addition of zinc in the superior position of the small Cu-7 decreases the incidence of pregnancy and that the small Cu-7 may be associated with higher retention rates than the standard Cu-7.


The clinical efficacy of the repeated transcervical instillation of quinacrine for female sterilization. The safety and efficacy of the repeated transcervical instillation of quinacrine hydrochloride in a suspension of 5 ml of 2% Xylocaine was evaluated in 200 patients. All instillation procedures were performed during the proliferative phase of the menstrual cycle: the second instillation was made in the first menstrual cycle following the initial instillation and the third and last instillation at 6 months after the first. None of the patients used any adjunctive contraceptives. Follow-up visits were scheduled at 6-month intervals after the last instillation. The potentially serious complications following the instillation were four cases of cortical exitation, and one case of acute adnexitis. The second instillation was not performed for 16.0% and the third instillation was not performed for 16.7% of the patients, for medical and/or personal reasons. Fifty-one pregnancies were reported, 41 (80.4%) before completion of the three instillations. The results of this study show that the instillation schedule used is unsatisfactory for widespread use. Additional studies are currently being conducted to evaluate the use of an adjunctive contraceptive up to the time of the third instillation in order to reduce the high pregnancy rate.


Fetal heart rate response to maternal hypotension during amniocentesis. Ten cases of severe fetal heart rate deceleration following maternal vaso-vagal reflex during amniocentesis were presented. The good outcome of these fetuses suggests that this transient phenomenon has no adverse effect on the fetus. This type of FHR response should be recognized so as to avoid unnecessary Caesarian sections done because of severe FHR bradycardia following amniocentesis.


Non-acceptance of puerperal sterilization. A study. This is an analysis of reasons given by 800 women who were hospitalized for deliveries at the Government Kilpauk Medical College Hospital in Madras for refusing puerperal sterilization. The cases are analysed by socio-demographic characteristics, age and parity. The most common reasons given were "inability to do manual work after the operation", "husband objects", and "necessity for prolonged rest". The author suggests ways of dealing with these barriers to sterilization.


Uterine rupture and vacuum extraction.

Uterine rupture and vacuum extraction. Eleven uterine ruptures occurred among 11415 vacuum extractions, and these cases are analysed. Cervical tears extending into the lower uterine segment or into the broad ligament occurred in 4 cases, lower segment tears in 6 and a fundal tear in 1. One patient died of respiratory failure. The possible cause of uterine rupture with vacuum extraction appear to be a combination of an unengaged fetal head, an incompletely dilated cervix and an undetected mild cephalopelvic disproportion especially in grandmultiparae, dehiscent or weak uterine scars, prolonged traction without progress, manipulations in shoulder dystocia and obstruction and excessive non controlled traction. Uterine rupture can be minimized by careful selection of cases, proper technique and early recognition of failure.


The use of the ultrasonic echo technique in examining the normal and pathological involution in the puerperium. The involution of the uterus in the puerperium was investigated after 50 normal, and 15 pathological pregnancies. By means of daily performed ultrasonic examinations the size and location of the uterus can be well determined, and the pace of the involution as well as the existing subinvolution can be diagnosed. It was found that in multiparas and in cases of uterine malformations the pace of the involution is somewhat slower, while in premature deliveries it is faster. By the help of this harmless and repeatable diagnostic technique the puerperal complications can be recognized and thus prevented.


Induced illegal abortions in Benin City, Nigeria. Abortions of all types constitute about 25% of our gynaecological admissions. In 8 months period between July 1974 and February 1975, 59 patients admitted into our unit with confirmed illegally induced abortions were studied personally by the author. It was found that most patients with this type of abortion were single, young schoolgirls. Most of the patients were 12 weeks pregnant or more at the time of the attempted termination. Most of the people who carried out these 'operations' by instrumentation were usually unskilled personnel. The complications in these patients which included one death are discussed. It is concluded that sex education including the use of contraceptive devices will help in reducing the high incidence of this 'social evil'. It is also thought that the liberalization of abortion laws in this country will also be helpful in reducing the complications associated with induced abortions.


Human chorionic somatomammotropin in normal pregnancy in an Asian population. Serial Human Chorionic Somatomammotropin (HCS) levels were measured during the last 10 weeks of pregnancy in 57 patients (375 samples). There was a gradual rise from 5.6 microgram/ml at the 30th week to microgram/ml at the 40th week, with a plateau during the last 4 weeks of pregnancy. No correlation could be found of HCS levels with parity, maternal age, maternal height, maternal weight, foetal weight and foetal sex. A weak but significant relationship was found between HCS and plasma oestriol (r = 2.4, p less than 0.001).


Fetal heart rate and fetal movements.

Fetal heart rate and fetal movements. Fetal Heart Rate (F.H.R.) in association with fetal movement was evaluated in 141 normal and pathological pregnancies. In the normal cases only 31% showed an acceleration of F.H.R. in association with fetal movement. The majority of the normal cases, 62%, did not demonstrate changes in F.H.R. in association with fetal movement. In the pathological pregnancies there were no characteristic changes in F.H.R. associated with fetal movement. It appears that F.H.R. acceleration associated with fetal movement cannot be used as an index for fetal well being.


Iron deficiency anaemia in a rural area in Nigeria. Evaluation of diagnosis and treatment with imferon. In 32 pregnant women with packed cell volume (PCV) of 30% or less (Hb 10 g/100 ml or below), iron depletion was present in 87.5% and megaloblastic changes in 84.5% as judged by bone marrow findings. Parenteral iron gave statistically significant rise in the PCV (p less than 0.001) in those who received iron therapy compared with control group with no iron therapy. Routine iron administration during pregnancy is therefore indicated in this population.


Acute carcinomatous myopathy associated with ovarian carcinoma. A case of acute, fatal, rapidly progressive pure myopathy associated with ovarian carcinoma is described. The first symptoms of carcinomatous myopathy occurred 8 months after surgical treatment and combined chemotherapy. The patient died 2 months after onset of myopathy. The clinical and pathologic findings of the myopathy are discussed.


The effect of miconazole on vulvo-vaginal candidosis in pregnant and non-pregnant women and their partners. Miconazole cream (Daktar vaginal cream 2%, Leo) given for 12-14 days was tested in 85 subjects with positive vaginal candida culture. Eighty percent of the patients were pregnant. Healing was observed in 93% of the cases. Treatment of the sexual partner did not influence the therapeutic effect.


Use of prostaglandin F2alpha in the management of missed abortion. Intravenous infusion of PG F2alpha was used to induce uterine contractions in 9 cases of missed abortions. Except for one case which was an extra uterine pregnancy, all of the patients aborted successfully. There were no major complications. The use of PG is suggested as a method of choice in the management of missed abortion specially in the second trimester.


Transplacental passage of diazepam following intravenous injection immediately prior to operative vaginal delivery. The early phase of diaplacental transfer of diazepam was studied in 39 women given the drug as a basic anaesthetic for operative vaginal delivery indicated by prolonged second stage of labour (9 cases), breech delivery (19 cases) and intrauterine hypoxia (11 cases). A total dose of 30 mg diazepam (Valium Roche) was injected intravenously over a period of 15 sec umbilical cord blood was collected immediately after delivery. Diazepam was extracted with diethyl ether and determined by gas chromatography. The concentration of diazepam in cord blood increased from greater than 5-250 ng/ml at 57-60 seconds to 48-1861 ng/ml at 90-100 seconds after completion of the intravenous injection. Thereafter a plateau seemed to be reached but the interindividual variation was still great with values ranging from 45-3034 ng/ml up to 360 seconds. Judged by Apgar score and the clinical course the neonates seemed to be unaffected by the medication administered to the mother.


Serum naphythylamidase isoenzymes during hormonal treatment. Electrophoretic and quantitative studies. Alterations in serum naphthylamidase isoenzymes were studied by electrophoretic and quantitative methods in women treated with oral contraceptives and women treated with naturally occurring conjugated estrogens for climacteric symptoms. In women treated with oral contraceptives the appearance of extra isoenzyme components was accompanied by a distinct and significant increase in the total serum naphthylamidase activity, whereas in treatment with conjugated estrogens no such increase was found. The result suggests that combined oral contraceptives and natural estrogens affect the serum naphthylamidase pattern in different ways.


Homotransplantation of the amniotic membrane for the treatment of congenital absence of the vagina. Homotransplantation of the amniotic membrane for the treatment of congenital absence of the vagina is described. Two years' follow-up with biopsy specimens revealed that amniotic membrane cells divide mitotically and activate vaginal epithelial proliferation of the squamous type and which respond to exogenous estrogens.


Perforation of the uterus by copper-IUDs. In about 5 000 copper-IUD insertions 5 cases of perforation were found. One woman had 2 devices inserted, both of them perforated. Copper-T was responsible for 3 corporal perforations and copper-7 for 2 corporal and 1 cervical perforation.


Twins, immuno-incompatibility and pre-eclampsia. A survey of 417 twin pregnancies is reported. Examination of the data revealed no difference in the incidence of pre-eclampsia referable to the zygosity of the twins. The findings thus lends no support to the hypothesis that immuno-incompatibility contributes to the aetiology of pre-eclampsia.


Calcified myoma of utero-ovarian ligament in a pubertal girl. A calcified myoma of the utero-ovarian ligament was encountered in a 13-year-old girl. The age of the patient and the unusual location prompt this report of a possibly unique entity, and the pertinent literature is cited.


An enzymatic time/temperature device for monitoring the handling of perishable commodities. Kockums Chemical AB, Malmö, Sweden, has developed a biochemical indicator, the i-point TTM (Time-Temperature Monitor). The indicator responds to temperature in much the same way as perishable commodities in which loss of quality is directly related to the combined effects of the degree and the duration of storage temperature. Primary application of such indicators are expected for frozen, refrigerated and fresh food products. The device can be useful in monitoring the handling of various temperature sensitive noon-food products as well. Reacting at a relatively low rate under optimum temperture conditions and at accelerated rates as temperature rises, the indicator chemically integrates and accumulates both the length and degree of all temperature experiences. When pre-selected time-temperature limits have been exceeded a distinct irreversible color change will occur, indicating that action is required--the particular depending upon  the use being made of the commodity.


Interrelationships between histamine, prostaglandins, and cyclic AMP in gastric secretion: a hypothesis. Based on the current experimental evidence, a model is proposed for mutual interactions of histamine, prostaglandins, and cyclic AMP in regulation of gastric secretion and pathogenesis of peptic ulcer. Histamine acting on H2-receptor-associated adenylate cyclase stimulates cyclic AMP formation and consequently secretion of hydrochloric acid in oxyntic cells. Prostaglandins (mainly E type) in another cell population stimulate cyclic AMP formation which may lead to formation of glycosaminoglycans and glycoproteins. Glycosaminoglycans and glycoproteins may have antisecretory and cytoprotective properties. In addition to this effect, prostaglandin endoperoxides may inhibit histamine-stimulated cyclic AMP formation in oxyntic cells.


Gastric adenosine triphosphatases: a review of their possible role in HCl secretion. In the search for gastric ATPases that might be related to the mechanism of HCl secretion, an interesting and rather unique K+-stimulated ATPase has been discovered. This enzyme is isolated from oxyntic cells and has been associated with the apical plasma membrane and/or tubulovesicular system. Membrane vesicles containing the K+-stimulated ATPase transport H+ into the vesicular lumen under the appropriate conditions of ATP, Mg2+, and KCl. This process can be measured by pH electrode, binding of certain metachromatic dyes to "energized" sites, or accumulation ratios of substances with appropriate pK values. Vesicular interior can be acidified to pH 3.5 or below. At the present time, it is difficult to distinguish between an electrogenic H+ pump and an electroneutral H+/K+ exchange mechanism. A hypothetical scheme for the gastric H+ secretory mechanism is proposed which fits much of the data from studies on the K+-ATPase, vesicular transport, and intact gastric mucosa.


Electrical events during activation and inhibition of gastric HCl secretion. A brief review of some of the relationships between the secretory rate and the electrophysiological characteristics of the canine and frog gastric mucosa is presented. There is no simple relationship between the potential difference (PD) and H+ rate in the in vitro frog gastric mucosa with standard Cl- bathing solutions. In contrast, with Cl- -free solutions (PD is inverted) there is a precise linear relationship between the PD and the H+ rate. This linear relationship is strong support for the concept that the H+ mechanism under these conditions is electrogenic. In the past it has been difficult rigorously to exclude the possibility that the inverted PD is due to a liquid junction potential between acid fluid in the lumina and the secretory bathing solution. Use of aminopyrine or imidazole essentially eliminates the possibility that the inverted PD is due to a liquid junction potential. Hence the linear relationship is solid evidence for the electrogenicity of the H+ mechanism. Recently we have shown that aminopyrine or imidazole can reverse thiocyanate inhibition of H+ secretion.


[Drug effects on blood pressure and heart rate in unanesthetized animals. (1). Effects of beta-blocking agents (author's transl)]. beta-Blocking actions of orally administered Kö 1400 and tiprenolol, new beta-blocking agents, were studied in unanesthetized rats and dogs, using a fall of blood pressure and an increase in heart rate produced by isoproterenol as a measure of beta-receptor activation. Blood pressure was recorded from the aorta of the dog and the caudal artery of rat via indwelling catheter, and heart rate of the dog was recorded by a cardiotachometer triggered by R waves of the lead II electrocardiogram. Mean resting blood pressure was 116 mmHg in rats and 93 mmHg in dogs, and heart rate was 99 beats/min in dogs. Isoproterenol (0.5 microgram/kg) was injected via indwelling venous catheter. Kö 1400, tiprenolol and propranolol inhibited the hypotension and tachycardia induced by isoproterenol at an oral dose level of 2 mg/kg or more. beta-blocking action in these preparations was found to be tiprenolol greater than Kö 1400 greater than propranolol. Pharmacological half life of tiprenolol was longer than that of propranolol, whereas that of Kö 1400 was shorter. No selectivity of beta-blocking actions was observed with all three beta-blockers. These findings are in agreement with the results obtained in isolated atrial and tracheal preparations of the guinea pig.


[Effects of a new adrenergic beta-blocking agent, Kö 1400 on the canine heart-lung preparation supported by a donor dog and the perfused hindlimb preparation of the dog (author's transl)]. Using the canine heart-lung preparation supported by a donor, effects of a new adrenergic beta-blocking agent, dl-1-(tert. butylamino)-3-[ (2-propinyloxy) phenoxy]-2-propanol hydrochloride (Kö 1400), on cardiac function, myocardial metabolism, and coronary circulation were studied and compared with those of propranolol. Effects of this substance on the peripheral vascular bed were also studied in the perfused hindlimb preparation of the dog. Kö 1400 produced a positive inotropic and chronotropic effect. Mechanical efficiency of the heart improved after Kö 1400, while it lessened after propranolol. A decrease in the coronary flow was observed in association with a slight increase in myocardial O2 consumption, indicating that the substance exerted a direct constrictive effect on the coronary vasculature. As a beta-blocker, Kö 1400 was found to be 2-3 times more effective than propranolol. The uptake of the free fatty acid (FFA) by the heart was increased by Kö 1400 and the myocardial redox potential improved. In the perfused hindlimb preparation, intraarterial injection of Kö 1400 resulted in a transient increase of the femoral blood flow for a decrease in the flow followed by a sustained decrease. With repeated administration a marked tachyphylaxis was observed.


[Studies on gamma-oryzanol. III. Influence of gamma-oryzanol on circadian rhythms of serum gastrin, 11-OHCS and gastric secretion in rats (author's transl)]. In the course of investigations of gamma-oryzanol in rats, circadian rhythms in antiulcerogenic action on gamma-oryzanol were observed. Circadian rhythms in several other parameters were investigated and correlated with the antiulcerogenic action. Pretreatment of rats with gamma-oryzanol 100 mg/kg, s.c., for 5 days tended to decrease serum gastrin levels and gastric secretion, and to increase serum 11-OHCS. Suppresive patterns of serum gastrin and gastric secretion following the pretreatment with gamma-oryzanol corresponded to the circadian rhythms in antiulcerogenic action of gamma-oryzanol. A complication of our results suggests that increment of catecholamines in rat brain after gamma-oryzanol plays a role in the above mentioned actions.


[Comparative study between dobutamine and other catecholamines in their effects on the cardiac contraction and rhythm (author's transl)]. Dobutamine (DOB) showed a dose dependent positive introphic effect on the right ventricular muscle of vagotomized Beagle dogs under pentobarbital anesthesia. Positve inotropic effects of other catecholamines (CA) decreased in the following order; isoproterenol (Iso, 115) greater than norepinephrine (NE, 15) greater than DOB (1) greater than dopamine (DA, 0.36). Brachiocephalic arterial flow was also increased by DOB with a positive inotropic effect in the right ventricular muscle. Though the positive inotropic effect of DOB was slightly decreased by pretreatment with phenoxybenzamine, such was almost completely antagonized by propranolol. Contractions of isolated cat papillary muscle driven by electrical stimulation (12 beats/min) were increased by CA in the following order; Iso (17) greater than NE (3) greater than epinephrine (2.5) greater than DOB (1) greater than DA (1 greater than). Activity of DOB to induce spontaneous contractions of papillary muscle was weakest among these CA. To determine the arrhythmogenic effect of these CA, coronary ligated Beagle dogs were employed, in which the anterior descending branch of the left coronary artery was ligated. Arrhythmogenic activity of DOB was weaker than other CA employed both in early stages and 48 hr after coronary ligation. DOB shows good potential for treating acute cardiac insufficiency and cardiogenic shock, as effects on heart rate and blood pressure are weaker.


[Clinical picture of lactate acidosis. 4: Clinical significance of lactate acidosis]. The diagnosis of lactate acidosis is complicated by the fact that lactate determination is not a routine method in clinical chemistry. In fact, lactate analysis is performed only in special laboratories. Even in greater clinics this method is not routinely performed in differential diagnosis of acidotic states. Various diseases are accompanied by a lactate emia or even by lactate acidosis. Anaerobic synthesis of lactate is an emergency reaction to supply minimum energy to tissues with insufficient oxygen supply. The main diseases complicated by increased blood lactate concentrations are shock, circulatory collapse, cardiac failure and peripheral circularoty disturbance. Additionally diabetes mellitus, septical infections, and-the most prominent situation-biguanide intoxications are complicated by an increase in blood lactate concentration.


Pathogenesis of cryptorchidism.

Pathogenesis of cryptorchidism. Cryptorchidism was induced experimentally by treating pregnant mice on the 14th day of pregnancy with 5 mg estrogen. Testes from cryptorchid and control newborn and adult mice were investigated with radioimmunoassay and electron microscopy. It was concluded that a normal Leydig cell function plays a decisive role in testicular descent. In cryptorchidism, Leydig cells at birth are atrophic. Testicular testosterone content is diminished as compared to controls. Ultrastructural alterations of Leydig cells observed in our experiments closely resemble those found in biopsies of cryptorchid patients. In male mouse offspring, prenatal estrogen injection induced not only a cryptorchidism but also Leydig cell atrophy and a permanently impaired function. Testosterone content is still significantly diminished after puberty. It is proposed therefore that an insufficiency of endocrine gonadal function of hypothalamo-pituitary origin occurring during intrauterine development is one of the main causes of cryptorchidism. An appropriate long-term therapy could diminish the high sterility rate.


The use of luteinizing hormone-releasing hormone in pediatric patients. Luteinizing hormone-releasing hormone (LH-RH), first synthetized in 1971; became soon available for clinical purposes and was immediately used as a diagnostic tool in children and adolescents as well as in adults. The first results of the LH-RH test in pediatrics were reported in 1972 and soon after discussed in meetings allowing comparison of the data from different groups. From this time, a great number of publications has been devoted to the diagnostic usefulness of LH-RH in the study of pubertal development and of pituitary-gonadal disorders in children and adolescents. Until recently, the theraeputic use of LH-RH has been restricted by the lack of availability of sufficient amounts, so that only few preliminary data have been reported in this field. Thus the scope of this review is mainly to present a critical survey of the data concerning LH-RH test in pediatric patients.


Differences in mouse interferons according to cell source and mode of induction. Mouse interferon induced by ultraviolet-irradiated Newcastle disease virus or polyriboinosinic-polyribocytidylic acid in T lymphocytes, B lymphocytes, macrophages, and primary mouse embryonic cell culture was studied. Irrespective of the inducer, interferons produced by T or B lymphocytes were relatively heat stable and of low antigenicity when reacted with antiserum against L-cell interferon (ALI), whereas interferons produced by macrophages and mouse embryo cells were heat labile and of high antigenicity against ALI. Mouse interferons induced by ultraviolet-irradiated Newcastle disease virus were separated into three components by chromatography on CH-Sepharose 4B. Interferons produced by T and B lymphocytes consisted primarily of component 1 (unbound fraction), whereas interferons produced by macrophages or mouse embryo cells consisted primarily of component 3 (eluted by 0.5 M NaCl). Component 1 was heat stable and of low antigenicity against ALI, properties characteristic of T- and B-cell interferon. Components 2 and 3 were heat labile and of high antigenicity against ALI, properties characteristic of macrophage and mouse embryo cell interferon. In contrast, interferon induced in mice sensitized with  BCG differed from these interferons induced in B cells, T cells, macrophages, and fibroblasts in being extremely acid labile and nonreactive against ALI.


Repression of heat-stable enterotoxin synthesis in enterotoxigenic Escherichia coli. Five different carbon sources were examined for their ability to control synthesis of heat-stable enterotoxin (ST) by enterotoxigenic (ENT+) Escherichia coli grown in either a defined medium containing four amino acids or a minimal salts medium. No ST activity was observed when D-glucose, D-gluconate, and L-arabinose were added separately to the defined medium, whereas glycerol and pyruvate decreased toxin levels. Similar results were obtained using a minimal salts medium, except with pyruvate, which did not support growth. Inhibition of ST synthesis by D-glucose was overcome by the addition of 3 X 10(-3) M cyclic adenosine 3',5'-monophosphate. Glucose repression of beta-galactosidase synthesis under conditions optimal for inhibition of ST synthesis was also reversed by exogenous cyclic adenosine 3',5'-monophosphate in the presence of the inducer isopropyl-beta-D-thiogalactopyranoside. The data suggest that control mechanisms for the synthesis of plasmid gene products of bacterial pathogens are similar to those exerted on the host chromosome.


Complement levels in pneumococcal pneumonia. Levels of complement proteins and functional activity of the alternate complement pathway were assessed in 39 patients with pneumococcal pneumonia. Mean levels of C3 and properdin and the functional activity of the alternate pathway in acute sera were significantly (P less than 0.05) below normal, whereas levels of components of the early classical pathway were normal. Although levels of factor B were in the normal range, they correlated significantly with C3 levels; there was no significant relation between C3 levels and C4 or C1q levels. The 19 patients iwth pneumococcal pneumonia and bacteremia had significantly lower mean values of properdin and factor B than the 20 patients without bacteremia, suggesting a more severe depression of the alternate complement pathway with bacteremia. During convalescence, complement levels were normal or elevated in most of the patients, but mean levels of properdin remained significantly below normal in bacteremic patients. Functional activity of the alternate pathway also remained below normal. These results indicate that there is a selective depression of the alternate pathway in patients with pneumococcal pneumonia, and they are consistent with the concept that the alternate pathway has an important role in host defenses in pneumococcal infection.


Cell envelope of Neisseria gonorrhoeae: relationship between autolysis in buffer and the hydrolysis of peptidoglycan. Neisseria gonorrhoeae readily underwent autolysis when suspended in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) buffer at alkaline pH values. Autolysis was inhibited by the addition of Mg2+ or other divalent cations. Autolysis was also suppressed at acid pH (pH 6.0). Suspension of cells in buffer was accompanied by the hydrolysis of peptidoglycan. The rate of peptidoglycan hydrolysis in HEPES buffer was maximal at pH 8.5 and was similar in the presence or absence of Mg2+. Therefore, divalent cation stabilization against autolysis is not mediated by inhibition of peptidoglycan hydrolysis. Peptidoglycan hydrolysis occurred in HEPES buffer (pH 6.0), but at a rate that was 50% of the maximum. Incubation of cells with chloramphenicol or rifampin before suspension in HEPES buffer (pH 8.5) partially prevented autolysis; under these conditions, peptidoglycan hydrolysis still occurred, but at a reduced rate. Old and new peptidoglycans were hydrolyzed at similar rates. Peptidoglycan hydrolysis results in solubilization of both the peptide and glycan moieties.


Delayed hypersensitivity in tumor-bearing mice. In vitro activation of "eclipsed" spleen cells. At various stages during the progressive growth of a transplanted sarcoma in BALB/c mice, the delayed hypersensitivity response to tumor antigen was determined using the food-pad swelling test (FPS) and the leukocyte migration inhibition assay (LMI). A close correlation was observed between the in vivo and in vitro assays. "Early" recognition of tumor antigen was detected 24 h after tumor inoculation by both techniques and this positive response was maintained until day 15. As the tumor grew larger, the delayed hypersensitivity response in vivo vanished, while the delayed hypersensitivity response in vitro disappeared about 3 days later. This suppression or "eclipse" of the anti-tumor cellular immune response was specific for the type of tumor used, and could be reversed in vitro by means of a low pH treatment of lymphoid cells.


Lack of association of the development of anti-sperm antibodies and other autoantibodies as a consequence of vasectomy. The possibility of autoantibodies--other than sperm antibodies--developing as a consequence of vasectomy has been investigated in 255 volunteers. During the first year after vasectomy no obvious increase was observed in the occurrence of any of these antibodies (rheumatoid factor, antinuclear antibodies and antibodies against smooth muscle, mitochondria, gastric parietal cell, thyroid microsomes and thyroglobulin). Ninety-nine of the patients were also examined for agglutinating and immunofluorescent antibodies to sperm to see if there was any relationship between the occurrence of anti-sperm antibodies and other autoantibodies. However, the prevalence of non-sperm autoantibodies did not differ in two nearly equal groups of patients with and without indications of autoimmune reactions to spermatozoa, respectively. Consequently the present results lend no support to the hypothesis that vasectomy could induce autoimmunity to other autoantigens than sperm-specific antigens.


Mixed lymphocyte culture responses in infertile couples. Twenty couples suffering from unexplained infertility have been tested in mixed leucocyte culture to determine if the wives were preimmunized against their husbands. The wives were tested against their husbands and four unrelated persons, and thymidine incorporation was measured on the third and fifth days of culture. No evidence was found that would indicate a preimmunization of the wife against her husband as an explanation for their infertility. A slight MLC stimulatory capacity was found in sera from infertile women.


A simplified determination of the urinary luteinizing hormone surge using the hemagglutination inhibition test. The hemagglutination-inhibition test for urinary luteinizing hormone (LH) has been modified so that the test for LH surge can be assayed in 1 day. A first morning voided urine specimen submitted to the laboratory can be analyzed in 5 hours by ordinary bench procedures. This modification involves a 20-fold reduction, as compared with the older procedure, of the duration of incubation of the urine-antiserum mixture. Another modification is the choice of aliquots which are adaptable to automatic pipetting. Only a single dilution of urine is necessary to determine LH surge for the anticipation of ovulation. Daily LH determinations on 12 normally menstruating women (av. cycle 28.18 days) reveal consistent patterns. For the 40 cycles studied LH surge occurred 14.29 days (av.) prior to the onset of the next menstruation and coincided with the rise in basal body temperature. One woman with an unusually high incidence of familial twinning showed two large LH surges in each consecutive cycle.


Hysteroscopic sterilization--a routine method? Our intention was to sterilize 50 women by means of hysteroscopic sterilization, and we were unsuccessful in 60% of these cases. All these women ran the risk of another pregnancy with hysteroscopic sterilization alone. Therefore--in our opinion--the method of hysteroscopic sterilization by thermocoagulation of the intramural part of the tubes is not yet a routine method, at least not before another modification of this method or instruments makes it possible to obtain better results.