Effect of pH on coenzyme binding to liver alcohol dehydrogenase. 1. The transient-state kinetics of ligand-displacement reactions have been analyzed. Methods based on this analysis have been used to obtain reliable estimates of on-velocity and off-velocity constants for coenzyme binding to liver alcohol dehydrogenase at different pH values between 6 and 10. 2. The rate of NADH dissociation from the enzyme shows no pronounced dependence on pH. The rate of NAD+ dissociation is controlled by a group with a pKa of 7.6, agreeing with the pKa reported to regulate the binding of certain inhibitory substrate analogues to the enzyme . NAD+ complex. 3. Critical experiments have been performed to test a recent proposal that on-velocity constants for the binding of NADH and NAD+ are controlled by proton equilibria exhibiting different pKa values. The results show that association rates for NADH and NAD+ exhibit the same pH dependence corresponding to a pKa of 9.2. Titrimetric evidence is presented indicating that the latter effect of pH derives from ionization of a group which affects the anion-binding capacity of the coenzyme-binding site.


The aromatic residues of bovine pancreatic ribonuclease studied by 1H nuclear magnetic resonance. 1. The aromatic proton resonances in the 360-MHz 1H nuclear magnetic resonance (NMR) spectrum of bovine pancreatic ribonuclease were divided into histidine, tyrosine and phenylalanine resonances by means of pH titrations and double resonance experiments. 2. Photochemically induced dynamic nuclear polarization spectra showed that one histidine (His-119) and two tyrosines are accessibly to photo-excited flavin. This permitted the identification of the C-4 proton resonance of His-119. 3. The resonances of the ring protons of Tyr-25, Tyr-76 and Tyr-115 and the C-4 proton of His-12 were identified by comparison with subtilisin-modified and nitrated ribonucleases. Other resonances were assigned tentatively to Tyr-73, Tyr-92 and Phe-46. 4. On addition of active-site inhibitors, all phenylalanine resonances broadened or disappeared. The resonance that was most affected was assigned tentatively to Phe-120. 5. Four of the six tyrosines of bovine RNase, identified as Tyr-76, Tyr-115 and, tentatively, Tyr-73 and Tyr-92, are titratable above pH 9. The rings of Tyr-73 and Tyr-115 are rapidly rotating or flipping by 180 degrees about their C beta--C gamma bond and are accessible to flavin in photochemically induced dynamic nuclear polarization experiments. Tyr-25 is involved in a pH-dependent conformational transition, together with Asp-14 and His-48. A scheme for this transition is proposed. 6. Binding of active-site inhibitors to bovine RNase only influences the active site and its immediate surroundings. These conformational changes are probably not connected with the pH-dependent transition in the region of Asp-14, Tyr-25 and His-48. 7. In NMR spectra of RNase A at elevated temperatures, no local unfolding below the temperature of the thermal denaturation was observed. NMR spectra of thermally unfolded RNase A indicated that the deviations from a random coil are small and might be caused by interactions between neighbouring residues.


Properties of Bacillus subtilis ATP-dependent deoxyribonuclease. A purification procedure described previously resulting in electrophoretically pure Bacillus subtilis ATP-dependent DNAse has now been modified by adding a fractionation stage with Polymin P to permit large-scale isolation of the enzyme. It has been found that the enzyme molecule (Mr = 300000) consists of two large subunits with Mr 155000 and 140000. The purified enzyme has three activities: (1) DNAse on linear single-stranded and double-stranded DNAs (2) DNA-unwinding and (3) ATPase. Circular DNAs were not affected by the enzyme. Study of the dependence of these activities on temperature, pH, and ATP and Mg2+ concentrations has revealed two different states of the enzyme. At low ATP concentrations and alkaline pH, it showed chiefly nuclease action, degrading considerable amounts of DNA to small fragments five residues long on average. At higher ATP concentrations and neutral pH (more physiological conditions) it predominantly unwound DNA. Simultaneously it cut preferentially one of the duplex strands to fragments more than 1000 residues in length. The results obtained suggest that the energy of the enzyme-cleaved ATP is mainly expended on unwinding rather than on degrading DNA molecules.


The kinetics of a purified form of 3-deoxy-D-arabino heptulosonate-7-phosphate synthase (tryptophan) from Neurospora crassa. 1. A method is described for the purification of a form of 3-deoxy-D-arabinoheptulosonate-7-phosphate synthase (tryptophan) that probably differs from that of the native enzyme. 2. The kinetics of the reaction catalysed by 3-deoxy-D-arabinoheptulosonate-7-phosphate synthase (tryptophan) shows that the reaction proceeds via a ping-pong bi-bi mechanism, with activation by phosphoenolpyruvate (P-Prv), the first substrate, and inhibition by erythrose 4-phosphate (Ery-P) the second substrate. At low substrate concentrations, KP-Prv is 0.1 mM and KEry-P is 0.13 mM. 3. The substrates phosphoenolpyruvate and erythrose 4-phosphate and the product inorganic phosphate can protect the purified enzyme against heat denaturation, whereas the inhibitor, tryptophan, has no effect, although it binds to the enzyme in the absence of other ligands. 4. Product inhibition by inorganic phosphate is linear non-competitive with respect to phosphoenolpyruvate (Ki, slope = 22 mM and Ki, intercept = 54 mM) and substrate-linear competitive with respect to erythrose 4-phosphate (Ki, slope = 25 mM). 5. The enzyme has an activity optimum at pH 7.3 and a tryptophan inhibition optimum at pH 6.4, Trp 0.5 is 4 microM. Inhibition by tryptophan is non-competitive with respect to phosphoenolpyrovate and substrate-parabolic competitive with respect to erythrose 4-phosphate. 6. The role of the enzyme in metabolic regulation is discussed.


Structure of pyridine nucleotide transhydrogenase from Azotobacter vinelandii. 1. Pyridine nucleotide transhydrogenase of Azotobacter vinelandii purified by affinity chromatography consists of a mixture of polydisperse rods at neutral pH. No other structures are seen by electron microscopy. 2. At high pH (8.5--9.0) the rods depolymerize. Complete depolymerization can be achieved in 0.1 M Tris-Cl pH 9.0. The depolymerized enzyme has a molecular weight of 421000 (sedimentation equilibrium), its sedimentation coefficient s20, w = 15 S and its Stokes' radius Rs = 7 nm. Since gel electrophoresis in the presence of sodium dodecyl sulphate shows that transhydrogenase consists of a single polypeptide chain of molecular weight (54 +/- 2) X 10(3) it follows that the depolymerized enzyme has an octameric quaternary structure. We propose that this octamer serves as the functional monomeric unit ('unimer') from which the polymeric form of transhydrogenase is constructed. 3. Gel filtration and sucrose gradient centrifugation studies of cell-free extracts from A. vinelandii show the unimer to be the predominant active species.


Mouse-liver glutathione reductase. Purification, kinetics, and regulation. Glutathione reductase from the liver of DBA/2J mice was purified to homogeneity by means of ammonium sulfate fractionation and two subsequent affinity chromatography steps using 8-(6-aminohexyl)-amino-2'-phospho-adenosine diphosphoribose and N6-(6-aminohexyl)-adenosine 2',5'-biphosphate-Sephadex columns. A facile procedure for the synthesis of 8-(6-aminohexyl)-amino-2'-phospho-adenosine diphosphoribose is also presented. The purified enzyme exhibits a specific activity of 158 U/mg and an A280/A460 of 6.8. It was shown to be a dimer of Mr 105000 with a Stokes radius of 4.18 nm and an isoelectric point of 6.46. Amino acid composition revealed some similarity between the mouse and the human enzyme. Antibodies against mouse glutathione reductase were raised in rabbits and exhibited high specificity. The catalytic properties of mouse liver glutathione reductase have been studied under a variety of experimental conditions. As with the same enzyme from other sources, the kinetic data are consistent with a 'branched' mechanism. The enzyme was stabilized against thermal inactivation at 80 degrees C by GSSG and less markedly by NADP+ and GSH, but not by NADPH or FAD. Incubation of mouse glutathione reductase in the presence of NADPH or NADH, but not NADP+ or NAD+, produced an almost complete inactivation. The inactivation by NADPH was time, pH and concentration dependent. Oxidized glutathione protected the enzyme against inactivation, which could also be reversed by GSSG or other electron acceptors. The enzyme remained in the inactive state even after eliminating the excess NADPH. The inactive enzyme showed the same molecular weight as the active glutathione reductase. The spectral properties of the inactive enzyme have also been studied. It is proposed that auto-inactivation of glutathione reductase by NADPH and the protection as well as reactivation by GSSG play in vivo an important regulatory role.


The proton-translocating adenosine triphosphatase of the obligately anaerobic bacterium Clostridium pasteurianum. 1. ATP phosphohydrolase activity. 1. The cell-membrane ATP phosphohydrolase of vegetatively grown Clostridium pasteurianum was specifically Mg2+-dependent, but demonstrated significant activity with GTP, CTP and UTP. It displayed approximate Michaelis-Menten kinetics only in the presence of certain effectors (e.g. phosphoenolpyruvate, fructose 1,6-bis-phosphate) which decreased the Km for ATP (to below 2 mM) but also V, whilst extending to pH 5.8 the effective pH range of activity of the enzyme. 2. ATP phosphohydrolase activity of the membrane ATPase (BF0F1) was inhibited by N,N'-dicyclohexylcarbodiimide, butyricin 7423, Dio-9, 4-chloro-7-nitrobenzofurazan, efrapeptin, leucinostatin and quercetin, and to a lesser degree by aurovertin and citreoviridin. The enzyme was not inhibited by oligomycin, spegazzinine, tributyl tin, triethyl tin or venturicidin. The soluble ATPase (BF1) component differed in not being inhibited by N,N'-dicyclohexylcarbodiimide, butyricin 7423 or leucinostatin. 3. The ATPase (BF0F1) complex and its soluble (BF1) component were separately purified. 4. Dodecylsulphate/polyacrylamide gel electrophoresis separated only four polypeptide components in the purified ATPase (BF0F1), with approximate molecular weights (+/- 10%) as follows: subunit a, 65 500; subunit c, 57 500; subunit da, 43 000; subunit fa, 15 000. The soluble (BF1 component contained only the three polypeptide subunits a, c and da. These were present in the BF0F1 preparation in the ratio 2 : 1 : 2; the contribution of subunit fa could not satisfactorily be quantified. 5. Subunit a was identified as the component binding 4-chloro-7-nitrobenzofurazan and subunit fa as the component binding N,N'-dicyclohexylcarbodiimide. The ATP phosphohydrolase activity of the membrane ATPase was not activated by trypsin treatment and the ATPase (BF0F1) contained no trypsin-sensitive inhibitor protein subunit. 6. Purified ATPase (BF0F1) was incorporated into artificial proteoliposomes which demonstrated ATP-dependent enhancement of 8-anilinonaphthalene-1-sulphonate fluorescence and ATP-dependent proton influx. These reactions were abolished by proton conductors (e.g. carbonylcyanide m-chlorophenylhydrazone) by valinomycin in the presence of a high external concentration of K+, or by N,N'-dicyclohexylcarbodiimide, butyricin 7423, Dio-9, 4-chloro-7-nitrobenzofurazan or leucinostatin. Oligomycin, tributyl tin, triethyl tin and venturicidin were not inhibitory. 7. When stripped of the soluble BF1 component, such ATPase-proteoliposomes demonstrated nil ATP phosphohydrolase activity and did not display ATP-dependent enhancement of 8-anilino-naphthalene-1-sulphonate fluorescence or ATP-dependent protein influx. All of these activities were restored by incubation of the BF1-depleted proteoliposomes with a purified preparation of the soluble BF1 component.


Alkylation of estradiol 17 beta-dehydrogenase from human placenta with 3-chloroacetylpyridine--adenine dinucleotide phosphate. 3-Chloroacetylpyridine--adenine dinucleotide phosphate is both active as a hydride acceptor and inactivates estradiol 17 beta-dehydrogenase. This coenzyme analogue behaves like an affinity label. The inactivation kinetics are discussed in relation to those observed with 3-chloroacetylpyridine--adenine dinucleotide. The pH dependence of the rate of inactivation, in combination with determination of the number of reactive cysteine residues, pointed to the alkylation of one cysteine residue/subunit. The stoichiometry was one molecule of dinucleotide per subunit and no cooperativity was detected. When 14C-labeled dinucleotide was used, the 14C label was found mainly in one peptide, accounting for 90% of the incorporated radioactivity, whereas in previous work it had been shown that 3-chloroacetylpyridine--adenine dinucleotide is an affinity reagent which labels three peptides.


Interaction of butene with human hemoglobin A. The binding of various alkanes by proteins was recognized years ago. We have studied the effect of butene (C4H8), a short-chain aliphatic hydrocarbon, on the functional properties of human adult hemoglobin. Under 1 atm pressure (100 kPa) butene decreased the affinity of hemoglobin (Hb) for oxygen (p50) by 45% without altering the cooperativity of ligand binding. This effect was independent of pH (from 7.0 to 8.0) and of ionic strength. The changes in the affinity of hemoglobin for oxygen were dependent upon the partial pressure of butene and evoked a saturating mechanism of the binding site(s). Mathematical simulation of the curve relating p50 to the concentration of dissolved butene allowed us to calculate the apparent association constants for one single binding site KHb = 10.4 mmol-1 and KHbO2 = 1.53 mmol-1 to Hb and HbO2 respectively. The larger binding of butene by Hb was confirmed by a 25% decrease in K1, the first association constant of oxygen to the tetrameric hemoglobin. It is concluded that butene is an allosteric effector of human Hb which acts most likely through hydrophobic interactions. It is postulated that the oxygen-linked binding site may be located at the alpha 1 beta 2 interface.


Biotransformations of glafenine in the rat and in man. The biotransformations of a therapeutic dose of the non-narcotic analgesic, glafenine, have been studied in the rat and in man. In the rat, the ester bond is extensively hydrolysed to give glafenic acid which is the major metabolite excreted in bile and in urine. Two minor pathways have been identified one leading by hydroxylation of the benzene ring of glafenine or glafenic acid in para of the amino-substituent to the corresponding phenols, the other, by oxidation of the quinoline nitrogen of glafenic acid, to its N-oxide. In vivo this N-oxide is partly reduced into the parent compound. Hydroxyglafenic acid is the product of both direct oxidation of glafenic acid and hydrolysis of hydroxyglafenine. The glyceric esters are conjugated as glucuro-ethers and/or sulfo-esters and the carboxylic metabolites as acyl glucuronides. The conjugation rate, high for glafenine, its phenol homologue and glafenic acid, is low for hydroxyglafenic acid and the N-oxide. The analogous urinary excretion patterns in man and in the rat suggest a similarity in the biotransformation of glafenine in these two species.


Methods for evaluation of urinary excretion parameters of alclofenac after I.M. administration of its water soluble lysine salt in man. The urinary excretion data after I.M. administration of water soluble lysine salt of Alclofenac were analysed by the techniques suggested by Niebergall, Wagner, Martin and Cummings. All methods used give similar estimates of both DU infinity and Kel with exception of the "Sigma minus" method when DU infinity obtained by "Rate Method" was utilized. Niebergall's method is preferred on the basis that it provides an accurate estimation of both DU infinity and Kel. These parameters, evaluated after administration of Alclofenac-lysinate, resulted in agreement with previously reported data obtained after administration of different pharmaceuticals of acidic drug.


Is glycine an inhibitory synaptic transmitter in the substantia nigra? Nigral tissue accumulated 14C-glycine by an energy-, temperature- and sodium-dependent mechanism; the transport process was inhibited by small neutral amino acids and had an apparent Km of 143 microM and Vmax of 787 nmol/g/min. Re-release of accumulated 14C-glycine was initially extremely rapid (40% in first 5 min) and subsequently accelerated by +40 mM K+ in a Ca2+-dependent manner. Intranigral kainate (0.25 microgram) lowered the levels of striatal DA (63%) and nigral GABA (25%) ipsilaterally, but not nigral glycine. Injections of glycine or strychnine (10--100 micrograms) into one SN induced slow ipsiversive or contraversive turning respectively. The evidence for glycine as a neurotransmitter in SN is discussed.


Interaction of estrogen and histamine during ovum implantation in the rat. The relationship between estrogen and histamine in the initiation of ovum implantation in the rat was studied. Histamine was found to augment the implantation response to suboptimal doses of estradiol in the ovariectomized pregnant rat maintained with Depo-Provera. The effect of histamine was found to involve both H1- and H2-receptors and to be inhibited by treatment with a combination of mepyramine and metiamide. Treatment with this combination of histamine antagonists during early pregnancy in the intact rat did not modify blastocyst attachment. It was concluded that the effect of histamine on the implantation response to exogenous estradiol was due to increased uterine blood flow and/or estrogen uptake but that such histamine-mediated effects were not essential for blastocyst attachment during normal pregnancy.


Clonidine mydriasis in the rat.

Clonidine mydriasis in the rat. Pupillary responses to clonidine (3--100 micrograms/kg, i.v.) and epinephrine (1--30 micrograms/kg, i.v.) were observed in anesthetized rats. Clonidine caused a dose-dependent mydriasis which was effectively antagonized by pretreatment with yohimbine (1.5 mg/kg, i.v.). Pretreatment with phentolamine (5 mg/kg, i.v.) was less effective in antagonizing this clonidine-induced mydriasis. Phenoxybenzamine (2 mg/kg, i.v.) was almost without effect. In contrast, both phentolamine and phenoxybenzamine blocked the pupillary dilation produced by epinephrine while yohimbine pretreatment resulted in no antagonism of epinephrine-induced mydriasis. These results suggest that clonidine-induced mydriasis in the rat is mediated by a central adrenergic inhibitory mechanism.


Modification of alpha-adrenergic responses of small arteries by altered PCO2 and pH. Closed-circuit television microscopy was used to measure in vivo small artery (75--140 microns) and vein (105--230 microns) diameters to determine if changes in tissue PCO2 and/or pH would alter the microvascular responses to norepinephrine. Sprague-Dawley rats were anesthetized with a combination of urethane (800 mg/kg) and alpha-chloralose (60 mg/kg). The cremaster muscle with intact circulation and innervation was suspended by sutures in a 60-ml bath which contained a modified Krebs solution (31 degrees C) that was buffered by Tris of bicarbonate. There were four groups of animals with different combinations of bath PCO2 and pH: (1) PCO2 less than 10 mm Hg and pH = 7.2, (2) PCO2 less than 10 mm Hg and pH = 6.9, (3) PCO2 = 60--70 mm Hg and pH = 7.2, and (4) PCO2 = 60--70 MM Hg and pH = 6.9. The maximal responses of the small artery and vein to norepinephrine were similar for the four groups. The artery sensitivity to norepinephrine was significantly lower for group 4 when compared to groups 1, 2 and 3, but there was no effect on small vein sensitivity. Thus, the combination of decreased pH and increased PCO2 reduces small artery sensitivity to norepinephrine in the cremaster muscle of the rat.


Changes in dopamine-mediated behaviour during one year's neuroleptic administration. Trifluoperazine (2.5--3.5 mg/kg/day) or thioridazine (30--40 mg/kg/day) were given in the drinking water to male Wistar rats for 12 months. Initial catalepsy and inhibition of spontaneous locomotion disappeared by 3 months and thereafter. Initial inhibition of stereotypy induced by s.c. apomorphine also disappeared by 3 months to be replaced by an enhanc-d stereotypy response after 6 and 12 months' drug intake. Drug-treated animals exhibited a greatly increased incidence of spontaneous mouth movements after 12 months' intake compared with control animals. Lower doses of both drugs (trifluoperazine 0.7--0.9 mg/kg/day; thioridazine 6--8 mg/kg/day) also initially suppressed behavioural responses but by 1 month and thereafter these animals were indistinguishable from controls. At 12 months, however, these animals also exhibited an increased incidence of spontaneous mouth movements. The data demonstrate a reversal of the initial dopamine receptor-blocking properties of trifluoperazine or thioridazine to be replaced by an enhanced response of cerebral dopamine systems while animals were still continuously receiving the drug.


Effect of catecholamines on oedema induced by inflammatory agents in the rat. The effect of intracutaneous adrenaline and noradrenaline (5 X 10(-12) and 5 X 10(-11) mol) was examined on the oedema (Evans blue dye leakage) response of rats to several inflammatory agents. The catecholamines reduced the oedema response to all agents tested except prostaglandin E1 (PGE1) which was significantly potentiated by noradrenaline (5 X 10(-11) mol), and a combination of bradykinin 5 X 10(-11) mol with PGE1 5 X 10(-10) mol which was not significantly affected by any dose of catecholamine. Adrenaline was more effective than noradrenaline in reducing oedema produced by 5-hydroxytryptamine (5HT) and histamine and by agents which release these amines (compound 48/80, dextran and antigen challenge with egg albumin in sensitized rats), but noradrenaline was more potent against bradykinin-induced oedema. The inhibitory effect of catecholamines against oedema produced by histamine and 5HT was abolished by a combination of phentolamine and propranolol. It was concluded that the oedema-inhibiting effect of catecholamines is due to alpha- and beta-adrenoreceptor mediated actions.


Albumin density gradient purification of canine hemopoietic blood stem cells (HBSC): long-term allogeneic engraftment without GVH-reaction. Long-term repopulation of the blood-forming organs of dogs, conditioned by wholebody X-irradiation (1200 R midplane dose), was achieved by transfusion of cryopreserved allogeneic blood mononuclear cells (MNC) without causing graft-versus-host-reaction (GVH-R). Donor and recipient dogs were DL-A identical, MLC-negative, no siblings, non-related. The blood stem cells (CFUc) were procured by a 3- to 4-hour continuous-flow leukapheresis. To increase the CFUc concentration in the peripheral blood, dextran sulfate (DS) was administered intravenously beforehand. About 1 x 10(10) MNC, among them about 1 x 10(7) CFUc, were collected and further segregated using a discontinuous albumin density gradient. Less dense cells were to be found in the upper part of the gradient (fraction 2). These cells included most of the CFUc, enriched by a factor of between 275 and 1730 compared to their concentration in the peripheral blood beforehand. After cryopreservation, these cells, when transfused into lethally irradiated dogs, completely repopulated the marrow and lymph nodes, caused no GVH-R and allowed long-term survival. These dogs received no immunosuppressive therapy, either before or after transfusion. More dense MNC were to be found in fraction 3; their transfusion caused a severe GVH-R, followed quickly by death. Fraction 4 was rich in lymphocytes and poor in CFUc. The transfusion of these cells produced a selective plasma-cell hyperplasia of the lymph nodes but failed to repopulate permanently the marrow. The reappearance of the different cell lineages in the marrow and in the peripheral blood after conditioning and transfusion of these cells produced a selective plasma-cell hyperplasia of the lymph nodes but failed to repopulate permanently the marrow. The reappearance of the different cell lineages in the marrow and in the peripheral blood after conditioning and transfusion of the segregated MNC is described in detail.


Inhibition of the NADP-linked glutamate dehydrogenase from Trypanosoma cruzi by silver nitrate. The purified NADP-linked glutamate dehydrogenase from Trypanosoma cruzi was strongly inhibited by silver nitrate. The inhibition was reversed by reduced glutathione, and was modified by the presence of the substrates during preincubation of the enzyme with the inhibitor.


[Preclinical study of the safety of a new tranquilizer, phenazepam]. Chronic toxicity of the benzodiazepine series tranquilizer phenazepam (5 and 50 mg/kg) administered per os for 1.3 and 6 months was studied in experiments on white rats, guinea pigs and dogs. It has been shown during varying time of observation that the drug neither decreases weight gain in the animals nor adversly acts on the blood or produces local irritating action. Histopathological changes in some internal organs proved irreversible. Administered at varying periods of pregnancy, phenazepam at an oral dose of 100 mg/kg exerted no embryotoxic or teratogenic action.


[Homovanilic acid accumulation in the brain of rats as a possible cause of the inhibitory action of psychotropic substances on behavior and motor activity]. In experiments on rats injection of haloperidol and levopromazine at growing doses (1--25 mg/kg) resulted in inhibition of the orienting-motor activity and catalepsy accompanied by accumulation of homovanilinic acid (HVA) in the forebrain. The dopaminergic drug apomorphine antagonized the inhibitory effects of the neuroleptics on behavior and simultaneously blocked the accumulation of HVA in the rab brain. Combined injection of haloperidol and probenecid potentiated catalepsy induced by the neuroleptic and led to a much higher level of HVA. levopromazine combined with probenecid elicited a deep inhibition of motility and myorelaxation. Bilateral microinjection of HVA at a dose of 50 micrograms into the caudate nucleus of the rabbit brain produced a clear-cut inhibition of the orienting-motor activity. It is suggested that the inhibitory action of the neuroleptics on behavior is partially mediated through accumulation of HVA by CNS synapses, i. e. HVA possesses its own neurotropic activity.


[Effect of preparations that alter catecholaminergic processes on the serotoninergic syndrome of head twitching in mice]. Specific neuroleptics suppressed head twitching (HT) in mice, provoked by d,1-5-hydroxytryptophane (200 mg/kg i. p.) in doses starting from 0.00025 mg/kg (spiroperidol). L-DOPA and piribedil inhibited HT in doses from 25 and 50 mg/kg respectively, whereas apomorphine and d,1-amphetamine in doses of 1 up to 10 mg/kg exerted ambivalent activity. HT was significantly attenuated by clonidine in doses from 0.25 mg/kg, whereas by noradrenaline, isoprenaline and naphthizine, injected into the brain ventricles, in doses of 1 microgram, and 0.025 microgram per mouse respectively. Destruction of brain catecholaminergic neurons by intraventricular 6-hydroxydopamine (50 micrograms per mouse) caused a strong enhancement of HT. However, partial protection of the adrenergic (but not the dopaminergic) neurons by pretreatment with desipramine or similar drug AW 15,1129 eliminated the protective effect of 6-hydroxydopamine. It is concluded that there is the dopaminergic link in the mechanism of HT and that the stimulation of the central adrenoreceptors inhibits HT.


Catecholamine reward pathways and schizophrenia: the mechanism of the antipsychotic effect and the site of the primary disturbance. Two catecholamine-containing pathways, the locus ceruleus system and the dopamine neurons arising from the ventral mid-brain, may be involved in reward. Dopamine neurons function as a system for energizing the organism's responses and directing them toward significant environmental stimuli, but the functions of the locus ceruleus system remain obscure. It appears increasingly likely that neuroleptic drugs exert their anti-psychotic effects in acute schizophrenia by blocking dopamine receptors, although the time course of the effect suggests that the mechanism is more complex than a simple reversal of a neurohumoral imbalance. Evidence from postmortem studies suggests that, at least in the chronic state, dopamine turnover is not increased, but that there may be an increase in postsynaptic receptor density in some cases, including some patients who apparently had not received medication in the year before death. The evidence is consistent with Olds and Travis' conjecture that "counteraction of positive feedback processes subserving positive reinforcement mechanisms may be a key to control of certain psychotic episodes".


Brain endorphins: possible role in reward and memory formation. A role for enkephalin in the mediation of behavioral reinforcement is supported by several lines of evidence: i) central injections of enkephalin serve as reinforcement for self-administration behavior, ii) electrical stimulation of many enkephalin-rich regions serves as reinforcement for self-stimulation behavior, which is blocked by moderate doses of naloxone, and iii) long-term retention of a passive avoidance response is facilitated by immediate post-learning injections of methionine-enkephalin and morphine.


Changes in red cell oxygen release capacity in diabetes mellitus. Studies are summarized to indicate that diabetes is associated with a fluctuating disturbance in the oxygen release capacity of the erythrocytes. This disorder, present from the onset of the disease, is a consequence of excess hemoglobin AIc, and absolute or relative hypophosphatemia and acidosis that interfere with formation of the red cell metabolite 2,3-diphosphoglycerate. As a result frequent increases in hemoglobin--oxygen affinity are produced. Available evidence suggests that transient decreases in red cell oxygen delivery lead to dilatation of the venous part of the microcirculation associated with increased transcapillary plasma permeation. Combined with microrheologic alterations (increased red cell aggregation, increased blood viscosity, and decreased red cell deformability) these functional changes may over the years participate in the pathogenesis of the microvascular disease in diabetes.


Dissociation of the proliferative and effector non-H-2 minor histocompatibility-alloimmune responses. The first-set and second-set allotransplantation reactions against skin grafts and the primary and secondary proliferative graft-versus-host reactions in the popliteal lymph nodes were compared in both directions in a non-H-2 system (mouse strain combinations C57BL/10ScSnPh (further B10) and B10.C3H(40NX) further 40NX) differing at H-1 plus H-?). While 40NX recipients gave stronger reactions against B10 antigens in the allotransplantation reactions, the situation was reversed in the GVHR, B10 cells reacting more strongly against 40NX antigens. The findings of a dissociation between the mechanisms of allotransplantation reaction adn proliferative GVHR suggest that the genetic determination of the target antigens and the reacting lymphocyte populations are more complex at the minor histocompatibility systems than has been expected.


[Clinical and immunological studies on acquired heat contact urticaria (author's transl)]. A case of localized urticaria in an otherwise healthy young woman, produced only by direct contact of the skin with heat, is described. The minimal temperature of urtication was 44 degrees C (immersion of the forearm in hot water for 5 min). Redness and painful oedema immediately developed without reflex flare. Total serum IgG, IgA, IgM, IgE, complement factors C3 and C4, and alpha1-antitrypsin were in the normal range, whereas the C1-inhibitor level was slightly decreased. There was no evidence of circulating immune complexes in the serum. A skin test and a RAST with house dust were positive, but there were no signs of respiratory atopy. An attempt for passive transfer of heat urticaria into the abdominal skin of a rhesus monkey failed, but was successful for house dust. A treatment trial with ketotifen, a new, perorally acting anti-allergic drug, was poorly effective, but dexchlorpheniramine maleate, a classical antihistaminic, in a dose of 12 mg daily completely suppressed the swelling evoked by heat, but not the erythema, suggesting that other tissue or plasma factors than histamine may be involved in the mechanism of heat urticaria in this patient.


Perspectives on fertility control.

Perspectives on fertility control. The biologic, administrative and individual perspectives on fertility control are presented and discussed. The need to unite perspectives is demonstrated and the importance of adjusting human fertility to new physiologies and social demands is stressed.


Measuring contraceptive efficacy and side effects. To assess effectiveness of contraceptives, one reports all pregnancies during use. Multiple and single decrement life-table pregnancy rates computed from these reports constitute the fundamental measures of effectiveness in clinical trials. Minimally biased evaluation of a new method requires a randomized, double-blind (or its nearest semblance), multicentered study of sexually active women. New contraceptives are studied in special, volunteer populations. Evaluation of marketed contraceptives implies broader-gauged representation, larger numbers and longer time frames. Prospective studies, like clinical trials, should include several methods, and may use subsamples for case-control analysis of risk. Well-designed and executed multicentered, prospective studies, nevertheless, may represent specially selected populations. Risk-benefit analyses based on findings from selected populations require careful scrutiny. Although the most persuasive estimates of effectiveness derive from national, representative samples, moderately large samples in the USA have not been able to distinguish among products within a class. Formidable problems of recall are present in household surveys. In the United Kingdom, Europe and North America, analysis of risks and benefits may be enriched by better utilization of the data collection and record linkage resources of national health statistics systems.


Oral contraceptive hypertension and thromboembolism. Oral contraceptives (OCs) are a highly effective and widely accepted means of avoiding pregnancy, but they also worsen the major atherogenic cardiovascular risk factors to some degree in all women. Some OC users may suffer severe hypertensive episodes or massive hypertriglyceridemia with pancreatitis. Mild or severe adverse effects could eventually have serious consequences beyond the childbearing years. OC use would appear imprudent for women with a history of hypertension, gestational hypertension and a family history of hypertension. Special care is needed with blacks, diabetics and women with renal disease. OCs may also affect blood clotting, fibrinolysis and platelet adhesiveness. Also, histochemical and anatomical changes in blood vessels have been noted. Both may precipitate thromboembolic events while the OC user is still at the childbearing age and may also contribute to accelerated atherogenesis in subsequent years. There is a need for more specific guidelines for monitoring women on OCs for a worsening of their cardiovascular risk profile and changes in blood coagulation. Indications and contraindications for OC use in relation to the hazard of thromboembolic sequelae need to be more explicitly defined.


Tumorigenic aspects.

Tumorigenic aspects. This report presents a review of tumors, except those of pituitary, that have been reported to occur in women taking combined oral contraceptive preparations. Pathologic features, both gross and microscopic, and differential diagnosis are emphasized. Particular attention is given to tumors of the liver: focal nodular hyperplasia (hepatic hamartoma) and liver cell adenoma (benign hepatoma). The characteristic features of these usually distinctive lesions are illustrated, and an attempt is made to evaluate the significance of each with respect to oral contraceptives. Tumorigenic aspects relating to the uterus and the breast are briefly discussed.


Barrier methods of contraception: a reappraisal. In the last two years, there has been a gradual reawakening of interest in barrier methods and an increase in their usage by both men and women. This is in large part due to concern about the sometimes serious side effects reported for other contraceptive methods. The return to these techniques is particularly important, given the current epidemics of teenage pregnancy and veneral disease. One of the major problems in relation to barrier methods today is the accurate determination of their efficacy. There are very limited data with statistical validity available to judge the exact rate of effectiveness one might obtain using one of these techniques. The National Survey of Family Growth, conducted by the National Center for Health Statistics, showed a failure rate per 100 women of 16.7 for foam, cream or jelly and 15.9 for diaphragms (22). There is a great need for new and improved barrier methods of contraception. Numerous clinical studies are being set up to test spermicidal agents and vaginal sponges for the female, as well as such things as disposable condoms for males.


Intrauterine devices: medicated and nonmedicated. The main benefits of intrauterine devices (IUDs) are a lack of adverse systemic effects, excellent effectiveness, high continuation rates and the single act of motivation required for use. First year failure rates range from 2% to 3%, but decline steadily thereafter to a cumulative annual failure rate of less than 1% after six years. The risks of IUDs include increased blood loss, uterine perforation, pelvic infection and pregnancy-related complications. The incidence of perforation of the uterine fundus ranges from 1:1000 to 1:2500 insertions, while that of cervical perforation with the copper devices ranges from 1:600 to 1:1000. IUD use is associated with about a three-fold increased incidence of developing acute salpingitis in comparison with use of oral contraceptives and diaphragms. If pregnancy occurs with an IUD in place, there is a three-fold increased risk of spontaneous abortion, a ten-fold increased risk of ectopic pregnancy (5% of all IUD pregnancies) and a possible increased incidence of sepsis during the pregnancy.


Sterilization of women: benefits vs risks. Voluntary sterilization is the birth control method most widely practiced throughout the world. The last ten years have witnessed great improvements in techniques and perfection of innovations, explaining the important role that it now plays in the regulation of fertility. Different methods are examined and it is concluded that hysterectomy is the best, if medically indicated; conventional laparotomy is not justified unless required by concomitant intraabdominal pathology; minilaparotomy is mostly suitable postpartum; colpotomy is better left to specialists; laparoscopy is ideal for nonpregnant patients; culdoscopy is a relic of the past; and hysteroscopy, although still experimental, may be the way of the future. The advantages of voluntary sterilization lie in its remarkable and immediate efficiency, freedom from ongoing motivation, the convenience of a one-time operation, the absence of side effects and the reduction of total costs. Its disadvantages are the complexity of any surgical intervention for a woman, its indisputable finality, its uncertain legality and the risks inherent in any operation. Hysterectomy and tubal ligation are practically never fatal, so this argument does not influence the choice of either method. However, incidence of morbidity is higher following hysterectomies, which must therefore be justified. The balance is clearly in favor of voluntary sterilization for the woman who is convinced that the size of her family is complete.


Vasectomy: benefits and risks.

Vasectomy: benefits and risks. Bilateral occlusion of the vas deferens, vasectomy, is progressively becoming the method of choice for couples seeking permanent contraception at a younger age, with smaller families. They are apparently well-informed and view the procedure as a natural step. Vasectomy is an inexpensively performed office procedure that causes minimal disruption of routine and has a high degree of community acceptance. The risks of significant hematoma, infection, discomfort and other sequelae are within acceptable limits. Improved techniques will continue to reduce the small failure rate. Antibodies observed in half of the patients have not been linked to systemic disease, although they are a hazard for the one patient in 500 returning for a vasovasostomy. Refinements in microsurgery and availability of artificial insemination enhance vasectomy as the method of choice. Evidently, extending the minimum time of sterility confirmation permits detection of occasional recanalization from technical failures. Adequate screening of the couple's motivation and expectations can prevent the rare psychologic disturbances, the greatest risk with this procedure and a problem associated with all options. For the male, there is no competitive technique at this time. In a world striving for equal rights, where the female still carries the burden of temporary contraception, the simplicity and popularity of vasectomy for permanent contraception add the desired undertones of social equilibrium.


Postcoital contraception.

Postcoital contraception. The evolution of postcoital contraception has led to the development of emergency measures to be used following a single unprotected act of intercourse and to ongoing methods, such as the administration of a contraceptive steroid agent following every coital exposure. In emergency situations, the most commonly employed hormonal steroids are the synthetic, conjugated and natural estrogens, administered in large doses for five days. Recently, a combination of an estrogen and a progestin has been employed for the same purpose. A copper-bearing intrauterine device (IUD), inserted within seven days of coitus, has also been utilized with success. Progestins alone have been utilized as an ongoing method of postcoital contraception. Failure rates have been found to vary with the dosage, the specific progestin employed and the frequency of intercourse. The major role of postcoital contraception in the developed world appears to be as an emergency measure. Ease of availability, a high degree of efficacy and a low incidence of side effects are essential for patient and physician acceptance.


A reappraisal of abortion as a method of fertility control. Some of the factors which led up to the passing of the British Abortion Act in 1967 (which came into effect ten years ago in April 1968) are outlined. British health advantages of the liberal and legalized abortion policy are described, as are the attitudes and opinions since passage of the act. Reference is made to the findings of the British Committee on the Working of the Abortion Act which were reported in 1974. The use of abortion as the sole method of fertility control in certain countries of Eastern Europe is discussed.


Problems of implementation and consequences of the 1975 provisional law to liberalize abortion in France. The passage of the 1975 law to liberalize abortion in France was inevitable because of the inadequacy of the 1920 law. The new law was passed quickly at the end of 1974 and its rapid implementation on January 17, 1975, caught public health officials by surprise. Two benefits have resulted from the law's implementation: (a) abortion has become a medical act, reducing the incidence of illegal termination of pregnancy and its complications; and (b) it has limited the need to travel to more liberal countries for an abortion. Chief among the law's disadvantages are the nonapplication of certain of its sections, the absence of means to curb noncomplying physicians and a certain laxity in its application due to inadequate public knowledge of the risks involved in repeated abortions.


Biochemical basis for the selection of oral contraceptives. Many specific plasma proteins show dose-related changes when oral estrogens are administered. Large increases in concentration are seen in many important binding proteins, such as the sex hormone-binding globulin, transcortin, the retinol-binding protein, ceruloplasmin, and transferrin. A smaller group of plasma proteins are reduced in amount. These changes are related to altered rates of hepatic synthesis and secretion. As the overall effect of estrogen is one of increased protein synthesis, there is a reduction in the amount of plasma-free amino acids and in the pattern of distribution. Oral contraceptive (OC) users frequently show significant alterations in biochemical tests of vitamin status, at least some of which are related to alterations in plasma proteins. Other biochemical changes associated with OC use include a fasting hyperlipidemia, due mainly to increases in triglycerides, although there is often also a small increase in cholesterol. These changes are due primarily to increases in several lipoprotein fractions and are related mainly to the estrogen component. A deterioration in glucose tolerance occurs in many OC users and is probably induced by both estrogens and progestogens. There is evidence that certain clinical side effects of OCs, such as depression, are associated with specific biochemical changes.


Endocrine effects of oral contraception. Numerous well-documented endocrine effects of oral contraceptives (OCs) can be subdivided into two groups: (a) those relating to the hypothalamic and pituitary-ovarian system, the breast and the genital tract of the human female and (b) those related indirectly to laboratory testing of other endocrine glands. Benefits from OC use include marked decreases in the incidence of menorrhagia, benign breast disease, dysmenorrhea, iron deficiency anemia, premenstrual tension and ovarian cyst formation--all of which will be discussed only briefly. Of much concern are two suggested adverse effects of OCs: (a) the development of amenorrhea or menstrual disturbance associated with anovulation after discontinuing OCs, thereby possibly influencing the future fertility of former OC users, and (b) the suggested relationship between hyperprolactinemia and the development of micro- and eventually macropituitary adenomas. A review of the literature, along with our own findings, is presented.


The British retrospective and prospective studies. The clinician needs scientific evidence to enable him to advise patients efficiently and correctly. The sources of this evidence are listed and the part played by each discussed. The rarity of adverse effects from the use of oral contraceptive pills entails the need to make use of epidemiologic studies. The different types of study are briefly described, with examples from recent work carried out in Britain. The advantages and problems of interpretation associated with each are discussed. The present position taken by United Kingdom doctors working in the field of contraceptive practice is summarized.


Smoking, age and the pill.

Smoking, age and the pill. A brief outline is given of alterations in the normal physiology caused by cigarette smoking, aging and taking combined oral contraceptive pills. There are similarities in some of these effects. Evidence is considered regarding the health risks of using the pill and the relationship of these risks to the age of the woman and her smoking habit. This evidence is derived from recent epidemiologic studies carried out in Britain on some aspects of morbidity and the death rates ascribable to the pill. The implications for prescribing clinicians are discussed and indications are given for reducing the overall risks to life and health for pill users.


Interpretation of data by the clinician. The cardinal challenges to every practicing physician are to interpret clinical data correctly and to place them in proper perspective. Clinical investigations frequently lack the rigidly controlled conditions and the careful experimental designs usually found in preclinical animal studies, and this deficiency is partially attributable to the inherent complexities of clinical medicine. Consequently, a great deal of controversy results from conflicting interpretations, extrapolations and overextension of limited data that are often equivocal. More careful appraisal of data and increased awareness of the well-known pitfalls found in retrospective and prospective studies, in which biostatistical design and clinical relevance are often incompatible, are emphasized, and personal biases and the flagrant sensationalism expounded by the media are condemned. The clinician is cautioned to sift through the data, consider the benefit/risk ratio for each patient and then to subordinate the role of critical scientist and assume the role of physician, exercising good judgment in light of the existing evidence and the immediate problems at hand.


Selecting the optimum method of contraception for each patient. The factors relating to the choice of the optimal method of contraception are discussed, indicating the wide range of variables which must be evaluated. Religious, cultural, educational and social factors are considered, as well as age and family size. Facts elicited through discussion of general medical, family, obstetric and menstrual histories help to indicate the advantages and disadvantages of available methods. The influences of the doctor, relatives, friends, and publicity are shown to affect the patient's choice and indicate that the theoretical effectiveness of a particular method forms only one part of a complex situation.


Initial pill selection and managing the contraceptive pill patient. Contraceptive pill risk may be lowered and patient continuation rates increased by selecting a pill best suited to each patient's needs. Each contraceptive pill has a unique profile of activity which is dependent upon both the dosage and type of progestogen and estrogen. Patient characteristics, expecially menstrual history, are helpful in choosing an initial pill (which should be low in estrogen), and patient and family histories are also valuable in helping determine whether use of the pill is contraindicated. Patients with a history of sensitivity to their own hormones are more likely to experience side effects on contraceptive pills, as also are underweight women. Knowledge of individual pill activities and potencies can be helpful in selecting the initial pill and in changing pills when nonserious side effects occur.


Advantages and disadvantages of low-dose oral contraceptives. Whenever a medication is prescribed, the lowest effective dose should be selected when possible. This recommendation is particularly important when oral contraception is being considered. In the years since the pill became available, there has been a steady decline in the dosages used. Contemporary "low-dose" varieties are those containing less than 50 micrograms of estrogen. Although they provide equal reliability, low estrogen formulations contain only a fraction of the quantities used in the original varieties. Subjective side effects are reported far less frequently with lower dose pills, and metabolic effects are reduced. The only associated disadvantage is a tendency to more spotting and breakthrough bleeding in early cycles. No matter how low the dose, there will always be some women for whom this method is not suitable. Fortunately, the progestogen-only pill--a consideralby underused contraceptive measure--can often be highly acceptable.


The patient's choice in contraception.

The patient's choice in contraception. The patient's choice of a contraceptive method is seemingly influenced by six factors: (a) her attitude toward contraception, (b) the physician's choice, (c) her assessment of risk, (d) her motivation and compliance for reliable contraception, (e) her previous experience with contraception and (f) her partner's influence. Recommendations are made for dealing with three groups of patients who present particular problems: (a) the patient who insists on the pill despite the contraindications for her emphasized by the physician; (b) the patient on the pill who is aging; and (c) the patient or couple who chooses tubal ligation at whatever age and regardless of the number of children she has borne.


Teenagers and contraception.

Teenagers and contraception. Early sexual activity in young women has created new problems in contraception and gynecologic pathology for physicians. None of the existing birth control methods seems ideally adapted to the young: oran contraceptives, the only infallible method, may present adverse effects. Intrauterine devices may result in expulsion or infection. Diaphragms or spermicides are less effective and not always well accepted by young girls. The physician, however, must bear in mind that whatever inconvenience may result, birth control is always preferable to an unwanted pregnancy or to abortion. Given the seemingly growing incidence of veneral disease and of abnormalities of cervical cytology, physicians must exercise the utmost care and consider a birth control consultation by a young girl as a full medical act.


Psychological factors: counseling and motivation of the contraceptive patient. The stronger the desire to avoid conception, the more effective the contracepting. But motivation alone will not ensure success. Knowledge of all aspects of contraception is a prerequisite for optimal decision making, and such information and counseling should be readily available to all persons of reproductive age who are concerned about the use of medication, potential side effects and subsequent fertility. The counsel and opinion of physicians are sought with as much or greater frequency as their diagnostic or therapeutic skills. Yet, too often, the physician's attitude toward contraceptive use by others is colored by his own moral, religious or social philosophy. Subtle personality traits of the patient and attitudes of her partner control contraceptive disuse, misuse or rejection in most people. In the author's opinion, compliance with the use of contraception is directly related to the amount and intensity of counseling and the degree of motivation provided by the physician.


A follow-up of vasectomy clients in rural Bangladesh. A one-year follow-up of 585 vasectomy clients sterilized at vasectomy camps in Shibpur and Shalna in rural Bangladesh showed that almost half of the clients were dissatisfied with their vasectomies. The majority of dissatisfied clients stated that their primary reasons for dissatisfaction were: (a) their ability to work had decreased and (b) they had not received all of the incentives they had been promised. However, 58% of the matched nonvasectomized controls also felt that their ability to work had decreased in the last year. Only 2%--7% of the dissatisfied clients cited decreased sexual performance as their primary reason for dissatisfaction. Satisfied vasectomy clients most frequently cited: (a) the permanence of this method of birth control and (b) the incentives they received as their reasons for satisfaction.


Implementation of a successful outpatient laparoscopic sterilization program in Calcutta. This study presents data on 109 women undergoing laparoscopic sterilization by tubal rings at the Calcutta Medical Research Institute and Hospital. The average operating time was 15 minutes and the average stay in the recovery room was 4 hours. The overall complication rate was 18%, including surgical and early postoperative complications. All patients returned for follow-up. Long-term follow-up will be required to determine the rates of delayed complications and the efficacy of the procedure.


Aerobic and anaerobic cervical flora of healthy Thai women. The bacteriologic study of the cervical flora in 50 healthy women revealed that 70% harbored mixed aerobic and anaerobic organisms. Aerobic organisms alone were recovered in 15 women (30%). Polymicrobial organisms were found in all but five women (in these only one aerobe was isolated per patient). The majority of patients had 1-2 anaerobes, with more than two aerobes. The common aerobes were alpha-hemolytic streptococci, Staphylococcus epidermidis and lactobacilli. The most common anaerobes were Peptococcus asaccharolyticus isolated in 21 women (42%), P prevotii in 13 (26%) and Bacteroides in 10 (20%). These organisms are frequently found in pelvic infection, suggesting the pathogenic potential of the normal flora of the cervix.


International maternity care monitoring: results of a pretest. This report gives the preliminary results of a pretest cosponsored by the International Fertility Research Program and the International Federation of Gynaecology and Obstetrics. It includes data on 33 116 deliveries in 20 maternity centers in Latin America, Europe, Africa and Asia. The findings are organized around four themes: (a) family formation and reproductive history, (b) family health, (c) management of this delivery and (d) desired family size and family planning practices.


Sex differences in fetal sheep adrenal steroidogenesis. The formation of several steroids was determined in vitro in adrenals removed from 18 female and six male fetuses of 113-115 days' gestation and in two female and two male fetuses at near term (137-143 days). Samples were incubated with 14C-acetate and the formation of labeled steroids was determined by two-dimensional paper chromatography. Protein and corticosterone concentrations were determined by chromophore absorption and acid hydrolysis fluorescence methods, respectively. Tissue corticosterone concentrations were significantly higher in female (0.145 +/- 0.010 microgram/mg protein) than in male (0.083 +/- 0.010 microgram/mg protein) adrenal tissue at both stages, whereas corticosterone formation was similar in both sexes. Cholesterol formation was significantly higher in female (0.103 +/- 0.079 muM/mg protein) than in male (0.044 +/- 0.011 muM/mg protein) adrenals at both stages. Both testosterone and estradiol were synthesized at higher rates in female than in male adrenals (52% and 33%, respectively), whereas pregnanediol formation was 21% higher in the male. These results indicate that significant sex differences exist in the formation of various adrenocortical hormones by fetal tissues. The relevance of these findings to better survival of female premature newborns from respiratory distress syndrome in contrast with male, is discussed.


Maternity record: initial report on a national experience (Colombia). The International Fertility Research Program, in cooperation with the Government of Colombia, drew a random sample of urban hospitals in which obstetric deliveries take place. Data were collected on a sample of the deliveries. Hospitals were divided into six types: university, university maternity, social security and three sizes of general hospitals. These groups of hospitals are compared with respect to the proportion of high-risk patients admitted, intervention rates and perinatal mortality rates.


A follow-up of tubectomy clients in Bangladesh. Five hundred twenty-one tubectomy clients from two health centers and one sterilization camp were interviewed 1-4 years after their tubectomies to determine: (a) demographic characteristics, (b) reasons for selecting tubectomy and (c) long-term satisfaction. Essentially no differences in demographic characteristics or long-term satisfaction were observed between urban and rural clients or by the type of center where the tubectomies were performed. Means ages of the clients were 31-32 years; mean parity was 7. Fifty-four percent of the rural and 85% of the urban patients had used contraceptives before tubectomy. Ninety-five percent or more of the patients stated that they were satisfied with the tubectomy.


Müllerian aplasia with hypoplastic thumbs: Two case reports. Two cases of Müllerian aplasia associated with unilateral hypoplasia of the thumbs and skeletal spine deformities in two unrelated females are reported, and the pertinent literature is reviewed. Müllerian aplasia is frequently associated with skeletal spine deformities, but has not been reported to be associated with hypoplasia of the thumbs. Several heritable syndromes, including the hand-foot-uterus syndrome, are characterized by uterovaginal and distal extremity malformations but are not associated with skeletal spine anomalies. The two cases reported here represent a previously unreported constellation of anomalies.


Rheumatic heart disease diagnosed during pregnancy: a 30-year follow-up. One hundred one patients originally diagnosed as having rheumatic heart disease (RHD) during the years 1945-1948 were reevaluated in 1975 to determine the natural history of the disease. Twenty patients (19.8%) showed no sign of RHD. Of the patients with confirmed RHD, 56 (70.0%) had their original lesion confirmed, while 23 (28.8%) had developed additional valvular involvement. Pure mitral stenosis resulted in significantly lower mortality than all other valvular lesions, and congestive heart failure was the leading cause of death. Nineteen patients underwent cardiac surgery; the mortality in this group (52.6%) was not significantly higher than that in the overall RHD group (38.8%). False diagnosis of RHD during pregnancy is common. A more thorough evaluation of the "cardiac murmur of pregnancy" is advocated.


Uterine and peripheral hematologic profiles in molar pregnancy. The potential differences in hematologic profiles of blood samples drawn simultaneously from the right utero-ovarian vein and from the upper extremity were investigated in four patients with uncomplicated molar pregnancy in stable obstetric conditions. The patients had undergone no previous chemotherapy and were scheduled for total abdominal hysterectomies. The dominant abnormalities in uterine venous blood were prolongation of thrombin time; shortening of activated partial thromboplastin time; positive protamine sulfate test; and increase in coagulation factors II and VII, with a tendency to low values in factor V. Peripheral samples gave almost parallel results in all altered and normal tests, except in one case with very striking differences in factors II, V, VII and X. Several local and systemic influences are discussed. It is concluded that molar pregnancy seems to have important systemic mechanisms affecting the stability of the blood coagulation homeostasis, which act in addition to those at a local level.


Further experience with a capillary tube pregnancy test. Further evaluation of the nonrefrigerated capillary tube pregnancy test is presented. A 127-mm by 2-mm glass capillary tube containing lyophilized antibody to human chorionic gonadotropin and latex indicator particles is used. Urine is drawn into the tube by capillary action; then, the tube is tilted to allow the urine to dissolve the antibody and to suspend the latex particles. The test results are read after a one-hour incubation period. Test results were evaluated against histopathologic and clinical data. Special efforts were made to stress the false-positive rate of the test by tripling the number of negative cases and doubling the sample size. A 99.82% agreement, a 0.18% false-negative rate and a 0% false-positive rate were obtained. In 52 confirmed pregnancies, the more sensitive capillary test detected human chorionic gonadotropin, whereas the commercial tests did not. Human chorionic gonadotropin was also detected one week earlier than reported in the first series.


The significance of adrenal and ovarian catheterization in patients with polycystic ovary syndrome. A previously unreported series of 15 anovulatory hirsute women with polycystic ovaries made up this study. Each underwent left adrenal and left ovarian vein catherization. Blood samples were obtained from the ovarian, adrenal and peripheral veins to test the levels of testosterone, delta 4 androstenedione (delta 4A), dehydroepiandrosterone (DHEA), estradiol and/or total estrogens, before and after administration of human chorionic gonadotropin (ten cases) and adrenocorticotropic hormone (ACTH) (five cases). Following intravenous human chorionic gonadotropin administration, the only response seen was a slight rise in ovarian levels of delta 4A and DHEA; after intravenous adrenocorticotropic hormone stimulation, the adrenal levels of delta 4A, testosterone and DHEA rose significantly. Peripheral concentrations of testosterone and delta 4A were in the high normal range and DHEA levels were high normal or slightly above normal range.


Viral hepatitis complicating pregnancy: mortality trends in Saudi Arabia. The striking mortality in viral hepatitis associated with pregnancy, regularly observed in developing countries, has shown a significant decrease in Saudi Arabia during a period of unprecedented economic growth. However, the risk of fatal hepatitis in the pregnant Saudi woman remains approximately four times that for the nonpregnant woman. The explanation for the observed mortality trend is not apparent, but is unlikely to be the result of improved nutritional status of the population alone, or because of treatment of severe hepatitis with adrenal corticosteroids. Disseminated intravascular coagulation may be one factor that decisively influences the outcome of hepatitis in the pregnant woman.


The effect of medroxyprogesterone acetate on blood pressure. Twenty-four women (21 normotensive and 3 hypertensive) aged 16-35 years received 150-mg injections of medroxyprogesterone acetate (MPA) for contraception. Their blood pressure (BP) was measured under basal conditions by the same nurse before treatment and at 1-, 2- and 3-month intervals. Their mean BP fell from 124.1/79.4 to 119.8/74.6 mm Hg at one month (p less than 0.05 for diastolic pressure) to 117.0/74.9 mm Hg at two months and to 115.6/73.2 mm Hg at three months. When the normotensive patients were analyzed separately, their BP fell, but not significantly. Only one patient had a rise of 20 mm Hg systolic, but she remained normotensive. We conclude that medroxyprogesterone acetate does not raise BP.


Microadministration of ketamine during labor and delivery of Nigerian women. Carefully controlled intravenous administration of ketamine produced excellent analgesia and dissociative sleep in all patients in the active phase of labor. The introduction to delivery interval was significantly shortened (3.6 vs 6 hours) in our general population. Of the 50 patients studied, 49 were unable to remember if contractions were painful and 80% experienced and narrated dreams. The auditory component was consistently intact in 82% of the cases; 78% described the total experience as pleasant, while the rest found it unpleasant or uncertain. The one-minute Apgar score of 6.8 was not significantly different from that of the general population.


Ultrasound diagnosis of hematocervix following gynatresia in a case of uterus bicornis bicollis. The authors describe the ultrasound findings of a hematocervix associated with cervical atresia. They emphasize the diagnostic difficulties and the important role of ultrasound in such cases.


Euhypnos Forte (temazepam) in insomniac patients: a clinical trial in general practice. In an open evaluation, soft gelatin capsules containing 20 mg temazepam (Euhypnos Forte) were administrated in doses of 40--60 mg to 134 patients from twenty-two general practices. The patients selected by the general practitioners werethose known to require hypnotics but in whom conventional doses had proven unsatisfactory. Of the 134 patients who entered the trial, 104 preferred the higher doses of two or three capsules. Eighty-eight per cent of these patients found Euhypnos Forte to be 'very good' or 'good' and 69% expressed preference for Euhypnos over their customary medication. Eighty-eight per cent of these patients experinces no hangover effect.


Euhypnos Forte (temazepam) for resistant insomnia: a clinical trial. Seventy insomniac patients, previously unresponsive to conventional hypnotic dosage, were treated for seven nights with temazepam in 20 mg soft gelatin capsules (Euhypnos Forte). The patients adjusted the dose to suit themselves up to a maximum of 60 mg. Nineteen patients found that one 20 mg capsule suited them best in spite of previous lack of response to two 10 mg capsules, and were thus excluded from the final analysis. Out of the remaining fifty-one patients, thirty-five were best suited by 40 mg and sixteen by 60 mg of temazepam. Sleep was rated Very Good or Good by forty patients (78.4%) and significant hangover occurred in only four (7.8%), all of whom were on 40 mg. Adverse reactions were insignificant. Some observations by one author (CALM) on the significance of the results in the management of insomnia in general practice are included.


Unlinked control of multiple glucocorticoid-induced processes in HTC cells. HTC cell variants chosen for their lack of tyrosine aminotransferase (EC 2.6.1.5) (TAT) induction by glucocorticoids were tested for interrelated effects on other glucocorticoid responses: TAT induction by dibutyryl cyclic AMP (dBcAMP) +/- dexamethasone, glutamine synthetase (GS) induction, cyclic nucleotide phosphodieterase (PDE) suppression, inhibition of alpha-aminoisobutyric acid (AIB) uptake, inhibition of plasminogen activator (PA), and induction of mouse mammary tumor virus (MTV). Loss of TAT induction by steroid was accompanied by loss of TAT induction by dBcAMP and of PDE suppression by steroid. In addition, subclones of MTV-infected cells were examined for the effect of the virus on glutamine synthetase (GS) and TAT induction. The virus had no effect on their induction in wild-type cells and no effect on GS induction in the variants. One MTV-infected subclone from a TAT variant, however, showed significant return of TAT induction.


The origins of unconjugated bilirubin in bile. In order to investigate the origin of unconjugated bilirubin in bile, beta-glucuronidase activity in rat and human bile was determined at various pH. beta-Glucuronidase in rat and human bile had their optimum pH at 5.5 when phenolphthalein glucuronide and delta 1-azopigment were used as substrate, and at 6.0 when bile itself was incubated. In human and rat bile the hydrolysis was suppressed to a minimum at each physiologic pH. However, human bile shows remarkable hydrolysis in alkaline pH (7.5--8.0). On the other hand, when delta 1-azopigment was incubated in various buffers, several per cent of delta 1-azopigment were hydrolyzed non-enzymatically in neutral to alkaline pH. Thus, it was suggested that enzymatic and nonenzymatic hydrolysis contributes to the existence of unconjugated bilirubin in bile.


Serum glycoproteins in the liver diseases. VII. Further studies on the properties of desialylated glycoprotein binding activity in particulate fraction of human liver homogenate. Binding of desialylated alpha 1-acid glycoprotein by human liver particulate fraction exhibited a dependence on the presence of calcium chloride whereas Cu+, Mn+, Zn+ Fe+ and Co+ inhibited the binding. The other cations such as K+, Na+, Ba+, Mg+ or Pb+ were determined to be non-effective on the binding activity. The pH of the assay for binding was not critical in the range of 6.5 to 9.5. The binding process required the presence of terminal sialic acid on the particulate protein. Fifty nine per cent of binding activity in the original liver paticulate fraction were recovered in acetone powder. Extraction of the acetone powder with a buffer containing EDTA resulted in an increased total binding activity. After extraction with 1--10% Triton X-100, 60% of the activity were still detected in insoluble fraction.


Reduction by cimetidine of acute gastric hemorrhage caused by reinfusion of blood after exposure to exogenous acid during gastric ischemia in rats. The present study has been made in anesthetized rats to characterize conditions of exposure to acid and extent of ischemia which determine the development of gastric hemorrhage. Gastric hemorrhage occurred in rats subjected to shock during exposure of the gastric lumen to acid after the reinfusion of withdrawn blood. When the acid concentration was constant, bleeding was dependent on the degree of shock. When hemorrhage shock was constant, bleeding from the stomach appeared pH dependent, although this did not quite achieve statistical significance. Cimetidine 2 x 10(-6) mol kg-1 min-1 (30 mg kg-1 hr-1) and 1 x 10(-5) mol kg-1 min-1 (150 mg kg-1 hr-1) significantly reduced gastric hemorrhage whether given prophylactically before gastric injury or therapeutically after completion of gastric injury. Because protection can be demonstrated against an injury involving exogenous acid and after the injury has been established, it is probable that the effectiveness of cimetidine in these studies is independent of its antisecretory effects.


Fate of oral neutralizing antacid and its effect on postprandial gastric secretion and emptying. The fate and neutralizing efficiency of oral antacids (aluminum and magnesium hydroxides) as well as their effect on postprandial gastric function were quantified in 6 patients with duodenal ulcer disease. We employed a double-marker technique for measurement of gastric secretion and emptying and combined this with back-titration of the gastric samples and analysis of aluminum to trace the fate of antacid in the stomach and duodenum. These studies show that: (a) antacid therapy with aluminum and magnesium hydroxides significantly increases gastric secretion; (b) intragastric neutralization of gastric acid produces a significant and substantial decrease in net acid output (acid secreted minus acid neutralized), but the beneficial effects of neutralization are partially offset by incomplete intragastric formation of aluminum trichloride; (c) most but not all of the ingested antacid is utilized in acid neutralization in the stomach (average 78.6% in our 6 patients); and (d) antacid therapy does not modify the absolute rate of postprandial gastric emptying, but increases dilution of gastric contents, expanding the intragastric volume. Thus, the fractional gastric emptying rate declines, and this, in turn, should enhance antacid utilization by delaying its emptying.


Effect of ethanol upon gastric emptying. The effect of ethanol upon gastric emptying in healthy human subjects was studied by measuring the gastric emptying rates of three 750 ml meals, the osmolalities, energy densities, and pH of which were similar. Meal A, which contained 80 ml alcohol, emptied more rapidly than meal B, which contained 40 ml ethanol and 63.3 g dextrose; and meal B emptied more rapidly than meal C, which contained 126.6 g dextrose but no ethanol. The slower rate of emptying of the dextrose meal (C) was not due to an increased gastric secretory rate, as serial measurements of gastric pH were substantially and significantly higher with this than with the other two meals; nor was it due to a greater degree of duodenogastric reflux, as serial measurements of gastric bile acid concentrations were similar for the three meals. We conclude that the duodenal osmoreceptor mechanism is relatively insensitive to ethanol; that the relationship between energy density and gastric emptying rate does not hold in the case of ethanol; and that the gastro-oesophageal reflux which occurs in response to ethanol is not due to impairment of gastric emptying.


Placental clearance of lactate and bicarbonate in sheep. To determine the placental clearance of lactate and bicarbonate in sheep, the fetal side of an isolated cotyledon and the umbilical circulation of the total placenta were artificially perfused. The release and uptake of lactate and bicarbonate by the perfusion fluid and the fetomaternal concentration differences of these substances were measured. From these data, the clearance of lactate and bicarbonate was determined to be 0.9 (SE = 0.2) ml/h/g of placental tissue. The production of lactate by the placenta was calculated to be about 30 mumol/min, the placental permeability of lactate was evaluated to be about 1.3 ml/h/g of placental tissue. These results indicate that fetal concentration changes of lactate and bicarbonate due to placental transfer occur with a half time of about 4 h. It is concluded that the lactate and bicarbonate permeability of the syndesmochorial placenta of the sheep is about 20 times smaller than the placental permeability of the hemochorial placenta of the guinea pig. It seems unjustified to draw any conclusions from experimental data obtained in the sheep placenta for the transplacental acid-base balance between mother and fetus in human beings.


Effect of intestinal gamma-glutamyl transferase inhibitor on the amount of gamma-glutamyl metabolites in mouse. Experimental mice fed a balanced rodent chow, called LSM fodder, had markedly lower gamma-glutamyl transferase activity in the epithelium of intestinal villi then control mice fed wheat. After oral administration of gamma-14C-glutamyglycine, oxidized 14C-glutathione or gamma-glutamyl-p-amino-benzoate the amounts of gamma-glutamyl substrates and their metabolites in intestines, livers and kidneys of experimental mice were significantly lower than those in control mice. L-serine simultaneously administered with gamma-14C-glutamylglycine reduced the radioactivity of gamma-glutamyl substances in organs of the control mice. No differences in organ radioactivity of experimental and control mice were observed when some uniformly labeled with 14C amino acids were given. The obtained results are not in aggreement with hypothesis on a role of gamma-glutamyl transferase in amino acid transport.


[Antibiotics therapy in chronic bronchitis]. In chronic bronchitis antibiotic therapy is no causative therapy, because infection rarely is the beginning but the complication of an already existing airway disease. While sputum cultures are very limited in their value, blind chemotherapy of bacterial infections by substances like tetracycline, ampicilline, azidocilline or cotrimoxazole is allowed. In cases of additional obstruction a combination with corticosteroids sometimes may be useful.


[Risk of incompatibility reactions in the repeated use of contrast media in urography]. In a prospective study 260 urographies were evaluated regarding clinical compatibility. In altogether 11% of the patients mild incompatibility reactions were observed (heat sensation, urticaria, nausea, vomiting). 29% of the patients with known prior contrast agent incompatibility suffered from anaphylactoid symptoms mostly of a subjective type (heat sensation, nausea). There was no statistically significant difference in the frequency of objective symptoms (urticaria, vomiting) between patients with prior contrast media incompatibility (3%) and patients who had tolerated previous contrast media applications without symptoms (2%).


New decade brings new systems.

New decade brings new systems. Be it employee development, patient education, or community health education, the task of the hospital educator in the future will require broader skills and knowledge.


Production of heat-stable, methanol-soluble enterotoxin by Yersinia enterocolitica. Seven isolates of Yersinia enterocolitica serotype 0:8, recovered during an outbreak of gastrointestinal illness, were examined for enterotoxin production. All seven strains were enterotoxigenic in the suckling mouse model, and three of five isolates tested produced keratoconjunctivitis in the guinea pig eye model (Sereny test). Enterotoxin was detected in broth supernatant fluid after 12 h of incubation at 25 degrees C. The toxin was not inactivated by exposure to 121 degrees C for 30 min or by storage at 4 or -40 degrees C for at least 5 months. The enterotoxin was also acid stable and methanol soluble. Methanol extraction did not affect its heat stability or enterotoxic activity in suckling mice. These physical characteristics plus limited ultrafiltration studies suggest that the enterotoxin is a low-molecular-weight substance. Y. enterocolitica enterotoxin resembles Escherichia coli heat-stable enterotoxin in heat and pH stability, methanol solubility, and enterotoxic activity in suckling mice. However, its role in the pathogenesis of human diarrhea has not been established.


Killing of Neisseria gonorrhoeae by human polymorphonuclear neutrophil granule extracts. Neisseria gonorrhoeae was grown in vitro (on agar and in broth) and in vivo (in 10-day chicken embryos) and tested for its sensitivity to the bactericidal action of human neutrophil granule extracts. Under all conditions studied, type 1 and type 4 N. gonorrhoeae were killed equally well by dialyzed extracts of neutrophil granules (containing both azurophil and specific granule contents) and by the myeloperoxidase-Cl- - H2O2 bactericidal system. However, sensitivity to the bactericidal activity of granule extracts depended upon growth conditions and growth phase. Log-phase, egg-grown gonococci were the most sensitive; they were killed 100% by 250 to 300 micrograms of granule extract (60 min, 37 degrees C) per ml. N. gonorrhoeae grown on agar for 20 h (to stationary phase) were the least sensitive, being killed only 80 to 90% with 500 micrograms of granule extract per ml. Thus, susceptibility to granule extract of gonococci grown under the four conditions studied in this report decreased in the order: log phase, egg grown; log phase, broth grown; stationary phase, egg grown; and stationary phase, agar grown. Killing was time and temperature dependent; little killing occurred when incubations were done at 10 degrees C. Boiled granule extract had only minimal effects on N. gonorrhoeae viability. Addition of catalase (500 U/ml) to the granule extract bactericidal system did not protect; however, the same concentration of catalase completely inhibited the bactericidal activity of the myeloperoxidase-Cl- - H2O2 system.


Effect of different dietary carbohydrates on some hepatic dehydrogenases and total lipid during starvation and refeeding regimen. Lipogenic capacity of various dietary carbohydrates starch, glucose sucrose and lactose was tested during ad lib feeding and starvation followed by refeeding. Sucrose was found to have maximal effect on hepatic total lipid and the enzymes in the study followed by glucose and sago while lactose was found to be toxic. Starvation resulted depression in the activities of various enzymes. The enzyme activity inducing effect was again exhibited by sucrose diet during ad lib and restricted refeeding followed by starvation.


Purification and chemical characterization of the heat-labile enterotoxin produced by enterotoxigenic Escherichia coli. Heat-labile enterotoxin (LT) produced by a human strain of enterotoxigenic Escherichia coli (286C(2)) was purified to homogeneity from pH extracts of fermentor-grown cells by ultrafiltration, (NH(4))(2)SO(4) fractionation, hydrophobic chromatography on norleucine-Sepharose 4B, hydroxylapatite chromatography, and Bio-Gel P-150 filtration. Purified LT preparations exhibited biological activity comparable to that of cholera toxin in four bioassays specific for the two enterotoxins (Y-1 adrenal tumor cells, Chinese hamster ovary cells, pigeon erythrocyte lysates, and skin permeability test). The overall yield of LT protein was 20%, which represented a 500-fold purification over pH extracts. A native molecular weight of 73,000 was determined by gel electrophoresis. The toxin dissociated upon treatment with sodium dodecyl sulfate, pH 7.0, into two components with molecular weights of 44,000 and 30,000. Purified LT preparations were remarkably stable over a wide range of storage conditions, temperatures, and pH's. The biological activity was increased by incubation with trypsin and completely destroyed by pronase and proteinase K, whereas deoxyribonuclease I, ribonuclease, and phospholipase D had no effect. The amino acid composition of purified LT was quite different from that of cholera toxin. Neither carbohydrate nor lipopolysaccharide was present in purified preparations. The purification scheme appeared applicable to LT produced by other human and porcine enterotoxigenic strains, but reflected the amount of LT produced by each strain. These data show that LT and cholera toxin share many common chemical and physical properties, but must be purified by different techniques.


Mechanism of action of Yersinia enterocolitica enterotoxin. Enterotoxin derived from three clinical isolates of Yersinia enterocolitica was compared with the heat-stable enterotoxin of Escherichia coli. Both toxins were biologically active in infant mice examined at 2 h and in ligated rabbit ileal loops at 6 h. Neither substance, however, produced changes in ligated ileal loops at 18 h or in Chinese hamster ovary or Y1 adrenal tissue cultures. In addition, both Y. enterocolitica enterotoxin concentrated approximately 20 times by ammonium sulfate precipitation and ultrafiltration and a similarly prepared sample of E. coli heat-stable enterotoxin stimulated the activity of guanylate cyclase but not that of adenylate cyclase in infant mouse intestine. These findings suggest that the role of enterotoxin in the pathogenesis of intestinal Y.enterocolitica infection may be similar to that of heat-stable enterotoxin in E. coli diarrhea.


Evidence for pili-mediated adherence of Klebsiella pneumoniae to rat bladder epithelial cells in vitro. The possible role of pili in the pathogenesis of urinary tract infections caused by Klebsiella pneumoniae was studied in an in vitro mixture of a phosphate-buffered saline suspension of rat bladder epithelial cells and phosphate-buffered saline-washed K. pneumoniae. Nonpiliated and piliated populations derived from a single K. pneumoniae strain were obtained by controlling the total time of growth in broth medium. The piliated phase demonstrated a significant increase in adherence when compared to the nonpiliated phase. Incubation of the bacteria and epithelial cell mixture at 4 and 37 degrees C resulted in no differences in adherence; optimal adherence occurred at pH 5. Pretreatment of the bacteria with enzymes to destroy the pili resulted in a decrease in adherence, as did killing the bacteria by various means before adherence testing. Pretreatment of the epithelial cells with certain saccharides inhibited bacterial adherence. Finally, a 96% decrease in adherence was observed after coincubation of bacteria and epithelial cells with papain-treated antipili antibodies. Thus, it appears that pili on the surface of K. pneumoniae mediate attachment of the bacteria to rat bladder epithelial cells.


Purification and characterization of the CFA/I antigen of enterotoxigenic Escherichia coli. The fimbral colonization factor antigen CFA/I of enterotoxigenic Escherichia coli was purified and characterized. The initial purification step was release of these fimbriae from the bacterial cells by homogenization with a Waring blender. Common fimbriae and flagellar antigen were avoided by careful control of growth conditions and the use of a nonmotile (H-) mutant of the prototype strain H-10407 (O78:H11). The essential purification steps were membrane filtration (Millipore Corp.), ammonium sulfate fractionation, and negative diethylaminoethyl-Sephadex column chromatography. Yields were approximately 4.0 mg of CFA/I protein per g (wet weight) of bacteria. Purified CFA/I is a fimbrial molecule 7.0 nm in diameter and has an average molecular weight of 1.6 X 10(6), as determined by sedimentation equilibrium. CFA/I is a polymer of identical subunits of molecular weight 23,800 with an N-terminal valine, 37% hydrophobic amino acid residues, and 11 residues of proline per  mol. The purified antigen retains its morphology, antigenicity, and biological activity. Purified antigen retains its morphology, antigenicity, and biological activity. Purified CFA/I exhibits mannose-resistant hemagglutination of human group A, bovine, and chicken erythrocytes, as do CFA/I-positive bacteria. This was demonstrated by sensitizing latex microbeads with the purified antigen since cell-free CFA/I fimbriae do not hemagglutinate erythrocytes. Thus, CFA/I detached from the bacteria are monovalent; however, purified CFA/I antigen retains an affinity for the epithelial cells of rabbit small intestine and blocks adhesion of CFA/I-positive bacteria. These results demonstrate that purified CFA/I is a good candidate for use in an oral vaccine for immunoprotection against diarrhea caused by CFA/I-positive enterotoxigenic E. coli.


Fimbrial hemagglutinin in stationary and shake cultures of Bordetella pertussis. Bordetella pertussis produced hemagglutinin in stationary cultures; in cultures kept under constant shaking, hemagglutinin was found only during the first 48 h of incubation but not after 3 to 5 days. The type of medium had a pronounced effect on production of hemagglutinin. Strain differences in ability to produce hemagglutinin were also detected.


Murine lymphoma alkaline phosphatase: a cell membrane carcinofetal enzyme. Alkaline phosphatase (APase) has been shown to have a membrane-bound localization in the murine fetal thymus, in murine thymic lymphoma and in adult spleen. Since it was suggested from these previous experiments that the lymphoma APase might represent an embryonic function, a detailed biochemical comparison of the lymphoma APase with the fetal thymus, placenta, fetus and spleen APases was performed. The parameters investigated were pH optimum, activation, inhibition, heat inactivation, substrate ratios, Michaelis constant, and electrophoretic analysis in the presence and absence of neuraminidase with the substrates alpha-naphthyl phosphate, beta-glycerophosphate, and p-nitrophenyl phosphate. The results indicate that the lymphoma APase is very similar to the fetal thymus, placenta and spleen APases. Furthermore, these results lend support to the hypothesis that the APase activity which appears in thymic lymphoma might represent a derepressed embryonic function. Thus, the murine lymphoma APase may be termed a cell membrane carcinofetal enzyme.


Hemodynamic changes in hypertensive patients at rest and during physical exercise before and after acute i.v. administration of bufuralol-HCl or propranolol. The hemodynamic effects of 20 mg Bufuralol-HCl and of 15 mg Propranolol given to hypertensives i.v. at rest and under physical exercise conditions were examined. It could be shown that Bufuralol-HCl lowered the diastolic BP and PR at rest already in the acute experiment, contrary to Propranolol. Under physical exercise conditions the diastolic BP is lowered, the PR remains unchanged in spite of reduced CO. After exclusion of other possible explanations, Bufuralol-HCl may lower the diastolic BP acutely at least partly by inhibition of cerebral beta-receptors. A faster and better liquor diffusion could be the reason for these results. It can be assumed that the acute BP lowering effect is mediated by the same mechanism as the chronic effect of the other beta-receptor blocking drugs.


A double-blind study of melperone and placebo in hospitalized chronic alcoholics in postintoxication phase. In a double-blind study in chronic alcoholics melperone (Buronil) was shown to significantly improve muscular and nervous tension, emotional lability, somatization, ability to sleep, anxiety, depression, paranoid ideation and presumed ability to work, but had no effect on alcoholic craving. The results received from three rating scales, and the theoretical aspects of alcoholism are discussed.


[Surgery of the adrenals].

[Surgery of the adrenals]. With 131 operated patients in the University Hospital of Urology in Zurich the indication for surgery in the 4 most frequent groups of adrenal disease is discussed. The preoperative localization of the tumor is exact in 96% of all cases with the serum examination in different stage of the vena cava. The appearance of hypertensive and hypotensive crises during surgery can be prevented with an adequate alpha-adrenergic blockade and volume replacement before and during the operation. In the surgery of Morbus Cushing we prefer the bilateral, dorsal incision on the 11th rib, if we haven't an unilateral adenoma. In the cases of Conn disease the removal of the whole surroundings of the adrenal gland is additionally indicated. The prognosis of the carcinoma of surrenal glands is poor, the adrenalectomy is difficult with regard to the early infiltration, the illness is seldom stopped on account of the frequent metastatic formation.


[Plastic surgical reconstruction of cutaneous loss of the heel or sole of the foot with sensitive free flaps]. In cases of total cutaneous loss of the heel or extensive cutaneous loss of the sole of the foot, the plastic surgical reconstruction by use of classical pediculated flaps requires following: - an adequate physical condition and articulations capable of enduring prolonged bed confinement in a position which is frequently uncomfortable; - a minimum of two operating procedures. It should be kept in mind that these free flaps are always insensitive and therefore subject to recurrence of wounding and trophic ulcers, etc. In order to avoid these drawbacks the authors suggest the use of a sensitive free flap, in which the neurovascular bundle is anastomosed to the existing bundle in the foot, using microsurgical techniques. Amongst the possible sensitive free flaps the authors favor the use of a sensitive latero-thoracic free flap. A detailed description of this free flap technique, its advantages and disadvantages for use in this kind of problem is described in their article. A clinical case is presented.


Oxidative deamination of thialysine by snake venom L-aminoacid oxidase. Thialysine is oxidatively deaminated by snake venom L-aminoacid oxidase at alkaline pH. The oxygen consumption curves show a characteristic diphasic course: the quick uptake of half a mole of oxygen per mole of substrate, in aggreement with a typical oxidative deamination, is followed by a slow extra oxygen consumption. The first product of the reaction is the corresponding alpha-oxo-epsilon-amino acid, which spontaneously cyclizes to the internal Schiff base 5-6-dihydro-delta 3,1,4-thiazin-3-carboxylic acid (TZCA). This latter has been identified by its UV absorption spectrum, by some chemical reactions, by paper chromatography, and by the production of cystamine and glyoxylic acid after prolonged oxidation of thialysine followed by acid hydrolysis. The possibility of an alpha-beta elimination reaction giving rise to cysteamine from thialysine, coupled to the oxidative deamination, has been excluded.


Variable pH dependence of the myosin-ATPase in different muscles of the rat. For the histochemical demosntration of the Myosin-ATPase (EC 3.6.1.3) the pH of both the preincubation and the incubation medium was varied in steps of 1 within a range: 10.2 to 10.5 and 9.3 to 9.9, respectively. The optimum combinations of both pH values, defined as the ones providing most consistent contrast among the three major types of muscle fibers were determined in 9 different muscles of the rat. The spectrum of optimum combinations differs considerably from muscle to muscle. The reduction of the incubation pH by only 0.1 may drastically change the staining pattern. This probably reflects the unspecificity of the histochemical procedure as well as the plasticity of the ATPase systems. To cope with the lability of the myosin-ATPase the optimum pH values of both media should be determined for each muscle separately.


8-Mercaptoflavins as active site probes of flavoenzymes. Representative examples of the various classes of flavoproteins have been converted to their apoprotein forms and the native flavin replaced by 8-mercapto-FMN or 8-mercapto-FAD. The spectral and catalytic properties of the modified enzymes are characteristically different from one group to another; the results suggest that flavin interactions at positions N(1) or N(5) of the flavin chromophore have profound influences on the properties of the flavoprotein. 1. The 8-thiolate anion form of 8-mercaptoflavin has an absorption maximum in the region 520 to 550 nm epsilon approximately 30 mM-1 cm-1). This form is retained on binding to flavoproteins whose physiological reactions involve obligatory one-electron transfers (e.g. flavodoxin, NADPH-cytochrome P-450 reductase). In the native form these enzymes stabilize the blue neutral radical of the flavin. A radical form of 8-mercaptoflavin is also stabilized by these proteins. 2. The p-quinoid form of 8-mercaptoflavin has an absorption maximum in the range 560 to 600 nm (epsilon approximately 30 mM-1 cm-1). This form is stabilized on binding to flavoproteins of the dehydrogenase-oxidase class (e.g. glucose oxidase, D-amino acid oxidase, lactate oxidase, Old Yellow Enzyme). These same enzymes in their native flavin form stabilize the red semiquinone, and have a pronounced reactivity with sulfite to form flavin N(5)-sulfite adducts. These properties of the native enzyme, including the ability to react with nitroalkane carbanions, are not exhibited by the 8-mercaptoflavoproteins. 3. A group of flavoenzymes fails to conform strictly to the above classification, exhibiting some properties of both classes. These include the examples of flavoprotein hydroxylases and transhydrogenases studied. 4. The riboflavin-binding protein of hen egg whites binds 8-mercaptoriboflavin preferentially in the unionized state, resulting in a shift in pK from 3.8 with free 8-mercaptoriboflavin to greater than or equal to 9.0 with the protein-bound form.