Enhanced naloxone distribution to the brain by morphine pretreatment in mice. An increase in the disposition of naloxone to the mouse brain was observed for animals previously exposed to morphine. Compared to controls, mice receiving morphine sulfate (10 mg/kg, sc) 3 hr prior to naloxone had a 28% increase in naloxone concentration in brain (200 to 260 pmol of naloxone per g of brain) 10 min after 3H-naloxone-HCl (0.4 mg/kg, 11.0 micronCi/kg, sc) administration. Also, if similar morphine-pretreated mice received a second dose of morphine sulfate (1.0 mg/kg, sc) concurrent with 3H-naloxone-HCl, the morphine-induced enhancement of 3H-naloxone concentration in brain was unaltered. This drug-treatment protocol paralleled that used by others in pA2-analgesia assays to demonstrate sensitization to naloxone for morphine-pretreated animals. In prior (3 hr) morphine-treated animals, administration of 3H-naloxone-HCl (0.1 mg/kg, 33.3 micronCi/kg) iv resulted in an 11.0% increase in 3H-naloxone brain concentration after 1 min. Thus, the enhancement of naloxone brain concentration was independent of the route of naloxone administration. No enhancement of 3H-naloxone brain concentration could be seen 24 hr after morphine sulfate pretreatment (10 mg/kg, sc), a decline in the effect similar to that seen for morphine-induced sensitization to naloxone. Finally, when morphine pellet-implanted mice (75 mg of morphine base, 72 hr) were administered 3H-naloxone-HCl (0.4 mg/kg, 10.0 micronCi/kg, sc), only a 22.5% enhancement of 3H-naloxone concentration in brain was obtained, as opposed to a reported 8-fold increase in the potency of naloxone. Thus, although a number of similarities exist between the enhancement by morphine of naloxone concentration in brain and its sensitization to the antagonistic activity of naloxone, a quantitative correlation appears to be lacking between the two phenomena.


The metabolism and toxicity of halogenated carbanilides. Biliary metabolites of 3,4,4'-trichlorocarbanilide and 3-trifluoromethyl-4,4'-dichlorocarbanilide in the rat. In separate experiments, after repeated oral administration of 3,4,4'-trichlorocarbanilide (TCC) and 3-trifluoromethyl-4,4'-dichlorocarbanilide (TFC) to rats, the biliary metabolites of each were isolated and identified. The major TCC biliary metabolite was found to be 2'-hydroxy-TCC. This compound was isolated mainly from the nonconjugated and the glucuronide fractions. Other metabolites present in substantial quantities were 6-hydroxy-TCC and 2',6-dihydroxy-TCC mainly as glucuronides and 3'-hydroxy TCC mainly as the sulfate conjugate. Small amounts of 3',6-dihydroxy-TCC were isolated from each of the fractions. No unchanged TCC was found in the bile. Only traces of other metabolites were found, and no N-hydroxylated products were observed. The major TFC biliary metabolite was the glucuronide conjugate of 2'-hydroxy-TFC. The only other metabolite of TFC was 3'-hydroxy-TFC, which was the predominant metabolite in the sulfate-conjugated fraction.


O-Demethylation of p-nitroanisole by Escherichia coli. Stimulation by phenobarbital. Intact cells of the bacterium Escherichia coli ATCC 11229, can convert p-nitroanisole into p-nitrophenol. The presence of phenobarbital in the culture medium during growth of the cells results in an inhibition of bacterial cell mass and an increased ability of the bacterial cell to carry out the O-demethylation reaction. There was a linear relationship between the amount of product formed and the bacterial cell mass in the incubation mixture. Varying the substrate concentration gave a concomitant change in activity, defined as the amount of product formed per mg of bacterial cell mass per 150 min. The optimum temperature for the production of p-nitrophenol by the stimulated cells was 33 degrees C; 7.0 was the pH optimum.


Lung pH and pulmonary absorption of nonvolatile drugs in the rat. The effect of pH on pulmonary absorption of nonvolatile drugs was investigated in the rat. Krebs-Ringer phosphate solutions (pH 6.2 and 7.4), Krebs-Ringer pyrophosphate solution (pH 8.4), or an unbuffered salt solution containing a drug were administered through tight-fitting tracheal cannulas to anesthetized animals. After 3 min, the lungs were removed and assayed for the amount of drug that remained. Weak acids and a weak base were absorbed most rapidly at pH values at which the drugs were least ionized. For example, with the base procainamide, 36% of the dose was absorbed at pH 6.2 and 76% at pH 8.4. With the acid sulfisoxazole, 71% was absorbed at pH 6.2 and 55% at pH 8.4. Similarly, with p-aminosalicylic acid, 77% was absorbed at pH 6.2 and 40% at pH 8.4. In contrast to these results, compounds such as urea and amitrole, which remain completely nonionized over the pH range studied, showed no change in absorption rate when the pH was varied. The two weak acids and the weak base were absorbed from an unbuffered solution as though the pH at the site of drug absorption was between 6.2 and 7.4. The absorption rate for each weak electrolyte from unbuffered solution, when compared graphically with the respective absorption rates from buffered solutions, indicated that the pH at the site of drug absorption is about 6.6.


Comparison of the metabolism of parathion by a rat liver reconstituted mixed-function oxidase enzyme system and by a system containing cumene hydroperoxide and purified rat liver cytochrome P-450. The metabolism of parathion by a reconstituted mixed-function oxidase enzyme system (rat liver cytochrome P-450, NADPH-cytochrome c reductase, dilauroyl phosphatidylcholine, deoxycholate, and NADPH) or a cumene hydroperoxide system (cytochrome P-450, dilauroyl phosphatidylcholine, and cumene hydroperoxide) have been compared. The products formed on incubation of parathion with both systems were paraoxon, diethyl phosphorothioic acid, diethyl phosphoric acid, p-nitrophenol, and atomic sulfur. The apparent KM values for parathion for formation of paraoxon and diethyl phosphorothioic acid with the cumene hydroperoxide system were 55 and 39 X 10(-6) M, respectively. These KM values are not significantly different. When the reconstituted system was used, apparent KM values of 2.8 x 10(-6) M for formation of paraoxon and 3.9 x 10(-6) M for The formation of diethyl phosphorothioic acid and diethyl phosphoric acid were determined. These KM values are also not significantly different. covalent binding of the sulfur atom, released in the metabolism of parathion to paraoxon, to the proteins of the reconstituted system and to cytochrome P-450 of the cumene hydroperoxide system was also examined. With both the reconstituted system and the cumene hydroperoxide system approximately 65% of the sulfur released became bound to the proteins of these enzyme systems. The binding of the sulfur atome resulted in a progressive inhibition of the metabolism of parathion by these two systems.


Metabolism of haloforms to carbon monoxide. I. In vitro studies. Trihalomethanes (haloforms) were metabolized to carbon monoxide by a rat liver microsomal fraction requiring both NADPH and molecular oxygen for maximal activity. GSH alone did not serve as a cofactor; however, GSH in the presence of NADPH and oxygen produced an 8-fold increase in the metabolism of bromoform to CO. Similar results were obtained with other sulfhydryl compounds. The biotransformation of bromoform to CO was characterized with respect to time course, microsomal protein concentration, pH and temperature. The metabolism of haloforms to CO followed the halide order; thus, iodoform yielded the greatest amount of CO, whereas chloroform yielded the smallest amount. A KM of 6.78 +/- 2.71 mM was established for bromoform and the Vmax was 1.09 +/- 0.19 nmol of CO per mg of microsomal protein per min. The energy of activation for this reaction was 6.5 +/- 0.18 kcal/mol. Cytochrome P-450 was found to bind bromoform to produce a type I binding spectrum. Treatment of rats with phenobarbital or 3-methylcholanthrene increased the rate of conversion of bromoform to CO. Cobaltous chloride treatment of rats or storage of microsomal preparations at 4 degrees C reduced the rate of formation of CO from bromoform. SKF 525-A inhibited the conversion of bromoform to CO. These results suggest that haloforms are metabolized to CO via a cytochrome P-450-dependent mixed-function oxidase system.


Neuropsychological deficit in polydrug users. A preliminary report of the findings of the collaborative neuropsychological study of polydrug users. The Collaborative Neuropsychological Study of Polydrug Users performed extensive neuropsychological assessments on 15 polydrug users 3 weeks after their enrollment in each of eight polydrug demonstration programs. Fifty-six (37%) of these subjects exhibited neuropsychological deficit. This deficit was partially related to increasing age, poor education and premorbid medical risk factors. The deficit was also associated with extensive and intensive use of two classes of drugs: sedatives (sleeping pills and minor tranquilizers) and opiates (heroin and other narcotic drugs). Seventeen (26%) of a comparison group of 66 psychiatric in-patients and day patients also demonstrated age- and education-correlated neuropsychological deficit. For these patients impairment was also related to lifetime experience with antipsychotic drugs and (perhaps) with clinical diagnosis of schizophrenia. Although both polydrug users and psychiatric patients revealed serious psychopathology as measured by the MMPI, the pattern of the neuropsychological test findings suggested that psychopathology alone did not account for impairment. The 3 month follow-up which is in progress should delineate further the time course and enduring features of neuropsychological deficit among polydrug users, and may establish more clearly the relationship of sedative and opiate use to such impairment. Changes in psychopathological status of both polydrug users and psychiatric patients should also help to clarify the influence of this variable on neuropsychological findings.


Disposition of carbamazepine and its 10,11-epoxide metabolite in the isolated perfused rat liver. The disposition of carbamazepine and its metabolite, the 10,11-epoxide, was investigated in isolated perfused livers from control and phenobarbital-pretreated rats. The mean hepatic clearance of carbamazepine was 0.28 ml per min per g of liver corresponding to a mean extraction ratio of 0.16 and a mean intrinsic clearance of 0.33 ml per min g of liver. In phenobarbital-pretreated rats, there was an approximately 40% increase in the hepatic clearance due to a concomitant increase in the mean extraction ratio from 0.16 to 0.26. The carbamazepine-10,11-epoxide generated in the liver approached and attained its peak concentration more rapidly in pretreated than in control rat livers. In the latter group there was virtually no disappearance of the epoxide from the perfusate, whereas in the pretreated rat livers the perfusate concentrations declined after 90 min. The extremely low clearance of the epoxide generated by metabolism in control rat liver was confirmed by administration of the epoxide per se to the perfusion system. The mean extraction ratio of the epoxide in this situation was only 0.016 and it was unaffected by prior treatment of the rats with phenobarbital. However, the epoxids exhibited dose-dependent kinetics with a 3-fold increase in the extraction ratio when the administered dose was reduced from 500 to 166 microng. The observed induction of the hepatic clearance of carbamazepine is supportive of the involvement of this mechanism in the interaction of this drug with various other anticonvulsants in both children and adults.


Induction of hepatic aryl hydrocarbon hydroxylase and epoxide hydrase in Wistar rats pretreated with oral methadone hydrochloride. Methadone-HCl added to the drinking water of adult female Wistar rats for 4 weeks produced an increase in the aryl hydrocarbon hydroxylase activity of the hepatic microsomal fraction to 222% of control levels. No change was seen in epoxide hydrase activity. In contrast, when male rats were treated similarly, there was an increase in epoxide hydrase activity to 212% of controls with no change in aryl hydrocarbon hydroxylase activity. No such changes were observed when the subcutaneous route of administration or chronic, low-dose, intraperitoneal injections were used. There were no differences in hepatic cytochrome P-450 or protein concentrations in treated animals as compared to their respective control groups. Control studies were carried out with quinine sulfate in the drinking water to decrease water intake to the level of the methadone-treated group. No elevation in either enzyme activity occurred in this control group. Similarly, paired-feeding studies showed the elevation of enzyme activity to be due to the methadone, not food deprivation. The effects of concurrent therapy of methadone with phenobarbital sodium or 3-methylcholanthrene were compared.


Metabolism and disposition of trifluoperazine in the rat. I. A thin-layer chromatographic method for the measurement of trifluoperazine and its metabolites in rat tissues. A method has been developed for quantitative measurement of trifluoperazine and its metabolites, 7-hydroxytrifluoperazine and desmethyltrifluoperazine, in rat organs. Trifluoperazine sulfoxide could also be assayed, but it proved to represent a very minor part only of total biotransformation products in tissues. Alkalinized tissue homogenates were extracted with di-isopropyl ether. Following removal of the bulk of lipids, the compounds to be quantitated were separated by thin-layer chromatography and measured by ultraviolet reflectance photometry on the plates. In recovery experiments, the method proved to possess a high reproducibility. The sensitivity limit for quantitative determination was about 0.1 nmol per extract, and the limit of detectability was 0.025-0.05 nmol. The applicability of the method was shown by analyzing the tissues of rats that had received 12.3 micronmol of trifluoperazine, ip, per kg.


Probenecid metabolism in vitro with rat, mouse, and human liver preparations. Studies of factors affecting the site of oxidation. The metabolism of probenecid in vitro was investigated with rat, mouse, and human liver preparations. As in previous in vivo studies, metabolism in vitro was found to be limited to the side chain. This metabolism involved mono-N-depropylation and hydroxylation in the 2- and 3-positions (3-hydroxy probenecid was in part converted to the carboxy metabolite). Acyl glucuronide conjugation was minimal. The presence of multiple forms of microsomal oxidative enzymes was suggested by the effects of: a) phenobarbital pretreatment, b) drug-metabolizing enzyme inhibitors, and c) variations in ionic strength and buffer composition on the metabolism of probenecid.


Inhibitory effect of immobilization stress on depression of liver tyrosine aminotransferase and tryptophan pyrrolase by glucose feeding in rats. In alloxan diabetic rats a stimulatory effect of stress on the activity of liver phosphoenolpyruvate carboxykinase seems to be very likely. In intact animals the inhibitory effect of glucose feeding (15% glucose instead of laboratory diet and water) on the activity of liver tyrosine aminotransferase (TAT) and tryptophan pyrrolase was reconfirmed. Moreover, a reversal of this effect by immobilization for 2.5 h was observed. After a mean intake of 5.3 g glucose/100 g body weight during 16 h this reversal was only partial and after 3.4 glucose/100 g during the same time the glucose effect was abolished. Stimulation of both enzymes by corticosterone and of TAT by stress-induced release of glucagon may play a role in this reversal.


Purification of bovine thyroid peroxidase. Trypsin-solubilized peroxidase activity from beef subcellular particles was resolved by DEAE-cellulose chromatography into 5 fractions, which contained enzymatically active components that ranged in molecular size from 73,000 to 340,000 daltons. The most active fraction (mol wt, 92,000 by gel filtration) was further purified (59,000-fold overall) by chromatography on hydroxylapatite. This highly purified peroxidase preparation had an absorbance purity ratio (A410:A280) of 0.55 and oxidized iodide (I3-formation) and guaiacol at rates of 300 and 460 micronmol/min/mg, respectively, which were about 3 and 1 1/2 times, respectively, greater than any previously described preparations. The enzyme was contaminated with an inactive protein of equal size. The highly purified peroxidase preparation lost its activity within a few days even when stored at -15 C with iodide. Two of the other DEAE-cellulose fractions contained peroxidase components with estimated sizes (gel filtration) of 73,000, 96,000, and 98,000, which were further purified purified (1,600 and 15,600 fold) on hydroxylapatite. They were 1/4 to 1/40 as active as the highly purified preparation and also became increasingly labile on purification. The remaining two DEAE-cellulose fractions were heterogeneous mixtures of stable peroxidase components whose average molecular sizes (gel filtration) were 220,000, 300,000, and 340,000 daltons, and which were not amenable to further purification on hydroxylapatite. The ratio of guaiacol to iodide activity decreased from 3.0 in the particles to about 1.5 in the highly purified preparations. The turnover numbers of the purest peroxidase component (mol wt. 92,000) for iodide and guaiacol were very similar to those of highly purifed, commericial lacto- and horseradish peroxidases. The pH maxima for iodide oxidation were 7.4, 6.0, and 4.5 for thyroid, lacto-, and horseradish peroxidases, respectively, whereas guaiacol oxidation peaked at pH 7.0-7.8 for all three enzymes. On the basis of these results and the dissimilar molecular sizes reported for trypsin-solubilized thyroid peroxidase by several other investigators, it was concluded that the molecular size is primarily determined by the conditions of proteolysis.


Beta-adrenergic stimulation of cyclic AMP content and parathyroid hormone release from isolated bovine parathyroid cells. The effects of beta-adrenergic agonists and antagonists on cyclic AMP (cAMP) accumulation and parathyroid hormone (PTH) release from isolated bovine parathyroid cells have been determined. Beta-adrenergic agonists markedly stimulate cAMP production and PTH release with an order of potency (-) isoproterenol greater than (-)epinephrine greater than greater than (-) norepinephrine, suggesting a beta2-type adrenergically mediated process. Both effects are blocked by the beta-blocker propranolol with the strict stereospecificity expected for a beta-adrenergic response. Low calcium concentrations also stimulate cAMP accumulation, but the cyclic nucleotide response under these conditions is only 3% of that obtained with isoproterenol, raising the possibility that factors other than cAMP may control low calcium-mediated PTH release. The release of PTH by low calcium is also not blocked by propranolol, confirming the independence of the response to low ambient calcium from the beta-adrenergic receptor. These studies substantiate further the utility of the isolated parathyroid cell preparation for studying secretagogue-mediated alterations in cyclic nucleotides and hormone secretion. Isolated cells also also make feasible the direct identification of beta-adrenergic receptors in parathyroid cell membranes and whole cells.


Direct identification of beta-adrenergic receptors on isolated bovine parathyroid cells. The radioiodinated beta-blocker iodohydroxybenzylpindolol ([125I]HYP) has been used to identify directly and characterize beta-adrenergic receptors in isolated bovine parathyroid cells. [125I]HYP was bound rapidly and reversibly to isolated bovine parathyroid cell membranes. Scatchard analysis revealed a single class of binding sites with high affinity (4 X 10(10M-1) and low capacity (0.7 pmol/mg). Saturation analysis of [125I]HYP binding to intact bovine parathyroid cells suggested a site with similar affinity on whole cells and with a binding capacity of 5000-10,000 sites/cell. True dissociation constants (Kd's) for beta-adrenergic agonists and antagonists were in good agreement with activation constants (KA'S) and inhibition constants (KI'S) for effects on adenylate cyclase in membrane preparations. These constants also were in reasonable agreement with KA'S and KI'S previously shown for effects of agonists and antagonists on cAMP accumulation and PTH release in whole cells. This study shows by direct analysis that beta-adrenergic receptors exist on isolated bovine parathyroid cells, and that there is close coupling between receptor binding, effects on cAMP and hormonal release. This represents still another system in which [125I]HYP has been successfully used to study beta adrenergic receptors in membrane as well as intact cell preparations.


[Testosterone reserve capacity in prepubertal and juvenile testes after sugery for undescended testis]. Tests testosterone reserve capacity of 6--15 year-old boys was estimated after operative correction of testicular maldescensus by a maximal stimulation test. Subnormal plasma testosterone levels were found in only 2 out of 14 patients with bilateral and 4 with unilateral orchidopexy. Prepubertal boys with unilateral anorchia had normal basal testosterone values and a normal testosterone rise after stimulation. In prepubertal boys with bilateral testes atrophy there was observed a diminished rise after stimulation. The basal testosterone levels were normal. The testosterone basal levels of pubertal boys with unilateral anorchia or bilateral atrophy were subnormal and the stimuation of testosterone production was reduced. The testicular volume of patients without atrophy or anorchia after orchidopexy was normal in prepuberty. During puberta a progressive relative decrease of the testicular volume was observed as compared to normal development. In conclusion, the results demonstrate that the endocrine function in most patients with unilateral or bilateral orchidopexy is in the normal range--a regular puberty can be expected.


Estimation of pregnenolone synthesis in rat adrenal homogenates: some cofactor requirements: effects of stress, hypophysectomy, cortisone and ACTH. Pregnenolone synthesis was estimated in whole adrenal homogenates incubated in the presence of cyanoketone (2alpha-cyano-4,4,17alpha-trimethyl-androst-5-en-17beta-ol-3-one). The yield of pregnenolone depended on the type of incubation medium employed. Both Ca++ and bovine serum albumin (BSA) markedly stimulated the rate of pregnenolone synthesis as did  NADPH or NADPH generating system. Aminoglutethimide added in vitro inhibited cholesterol sidechain cleavage activity. Ether stress in vivo stimulated pregnenolone synthesis in vitro, and hypophysectomy of 24 hours duration resulted in a decrease. Cortisone administration for 8 days reduced the formation of pregnenolone by rat adrenal homogenates, an effect prevented by concomitant treatment with ACTH. Similarly, hypophysectomy of 8 days duration resulted in a marked diminution of pregnenolone synthesis and ACTH replacement reversed this effect. Changes in pregnenolone synthesis were paralleled by changes in corticosterone and total steroid production.


The effect of repeated immobilization on the level of plasma corticosterone and on the activity of several liver enzymes in rats. The effect of repeated stress on the level  of plasma corticosterone and on the activity of several target enzymes for this hormone in the liver was studied. In adult male rats immobilized for 2.5 hrs daily, on day 7 the response of both plasma corticosterone and hepatic tyrosine aminotransferase is modified: After similar increases immediately after immobilization as in aminals stressed for the first time, in the conditioned rats precocious decreases to initial values take place. Moreover, on day 4, 24 hrs after a third immobilization, there are increases arise partly at least as a consequence of diminished food intake, as shown by comparing them with data from pair-fed rats. Partial fasting leading also to slight increase of hepatic glucose-6-phosphatase activity constitutes an  important part of repeated stress with substantial impacts on metabolic processes.


Tyramine oxidase activity in needle biopsy of normal livers and diseased livers. Tyramine oxidase and UDP-glucuronyl transferase activities were determined in 52 diseased livers obtained by needle biopsy. 14 liver specimens were also subjected to acetyl CoA carboxylase determination. Tyramine oxidase level was elevated in livers with nonalcoholic fatty change or toxic hepatitis, and reduced in livers with fibrosis or chronic alcoholic injury. UDP-glucuronyl transferase activity was reduced in livers with severe parenchymal damage or hyperbilirubinemia. Acetyl CoA carboxylase activity decreased markedly in an active alcoholic cirrhotic liver, and was elevated in alcoholic fatty livers as well as in a liver with acute venous stasis.


Interactions of divalent cations and nucleotides with solubilized cardiac guanylate cyclase. Some properties of guanylate cyclase, which was solubilized from the rabbit heart washed particles by the treatment with Triton X-100, were investigated. The solubilized enzyme activity was stimulated by Mg2+ in the presence of low (subsaturating) Mn2+ (GTP is greater than Mn2+); under these conditions, Ga2+ was inhibitory. At subsaturating MnGTP and free Mn2+, the solubilized enzyme was markedly stimulated by MnGDP and MnATP; CaGTP on the other hand, was inhibitory. These results are consistent with the view that the particulate guanylate cyclase may exist in the cell as a metalloenzyme with tightly bound Mn2+ and that Mg2+ supports its catalysis while Ca2+ as well as nucleotides may exert regulatory effects on its activity.


Hepatic alkaline phosphatase isoenzymes: isolation, characterization and differential alteration. Although it is generally believed that hepatic alkaline phosphatase is localized to liver plasma membranes, 63% is present in the cytosol fraction after ultracentrifugation of rat liver homogenates. Divalent cation requirements, heat inactivation, pH optima, Km and chemical inhibition characteristics of partially purified alkaline phosphatase enzymes prepared from membrane and cytosol fractions suggested different structural forms. Furthermore, bile duct obstruction and ethinyl estradiol administration preferentially increased membrane-bound alkaline phosphatase activity, while cytosol activity was unaltered. In contrast, phenobarbital treatment decreased membrane-bound alkaline phosphatase and increased cytosol activity. These studies support the presence of two forms of hepatic alkaline phosphatase in rat liver which are regulated by different control mechanisms.


The interaction of phospholipid membranes and detergents with glutamate dehydrogenase. 1. The interaction of beef liver glutamate dehydrogenase with cardiolipin from both beef liver mitochondria and beef heart mitochondria, with phosphatidylcholine from both beef liver mitochondria and egg-yolk, and with beef brain phosphatidylserine was investigated by steady-state kinetic methods. 2. the phosphatidylcholine did not inhibit the enzyme under a wide range of conditions. The cardiolipins and phosphatidylserine inhibited the enzyme. The inhibition by these lipids was found to diminish with time if the lipids were prepared and the reaction was studied in either phosphate or Tris buffers, but in zwitterionic buffers these lipid brought about a rapid, reversible inhibition which remained stable with time for at least 150 min. 3. The kinetic type of the inhibition was difficult to determine because of variation between lipid sonicates. Complex mixed types of inhibition were found with cardiolipin, and with phosphatidylserine the inhibition approximated to a non-competitive interaction with Ki(app) values varying between (0.9-6.1) x 10(-6)M. 4. The extent of inhibition decreased with increasing pH  and with increasing ionic strength. Basic proteins, such as cytochrome c, show a higher affinity for the anionic membranes and can dissociate the enzyme-lipid complexes. Cosonicates of the cardiolipin and phosphatidylcholine inhibited the enzyme, the extent of inhibition increasing in proportion to the amount of acidic lipid. 5. Sodium dodecylsulphate causes a time-dependent inhibition of the enzyme. The kinetics of this effect and its variation with detergent concentration were studied. 6. The relationship of these observations to the structure and function of the enzyme is discussed. It is suggested that their apparent regulation of the enzyme by oestrogens and other small molecules is due to their binding in vitro at sites on the enzyme designed for binding cardiolipin, when the enzyme is functioning in vivo. The association of the enzyme oligomer in vitro may, for similar reasons, be an artifact.


The interaction of phospholipid membranes and detergents with glutamate dehydrogenase. 1. Both the anionic detergent sodium dodecylsulphate and the cationic detergent cetyltrimethylammonium bromide quenched the protein fluorescence of glutamate dehydrogenase. The anionic compound was more effective and brought about 50% quenching at a detergent concentration of 0.4 mM. The zwitterionic amphiphile, lysolecithin, did not quench the protein fluorescence and neither did the short-chain detergent n-hexylsulphonate, which under the range of concentrations examined (less than 1 mM) does not form micelles. 2. The zwitterionic phospholipid, phosphatidylcholine, did not quench the protein fluorescence but the anionic phospholipids, phosphatidylserine and cardiolipin, induced a reversible quenching of the enzyme fluorescence. These observations confirm the specificity of the phospholipid-enzyme interactions as deduced from the kinetic studies of the preceding paper. The degree of quenching brought about by the phospholipids decreased with increasing ionic strength and increasing pH and could be substantially reduced by basic proteins. An electrostatic contribution to the interaction is inferred from these results. 3. The binding of the anionic phospholipids to the enzyme is manifested in a further enhancement of the fluorescence of a 1-anilinonaphthalene-8-sulphonate-enzyme complex. The presence of substrates and allosteric effectors affect the interaction of the lipids with the enzyme as indicated by the magnitude of this increase in fluorescence. The enhancement of fluorescence of NADH when bound to the enzyme was not affected by the binding of the lipids. 4. The complex formed between the enzyme and phosphatidylserine/phosphatidylcholine can be solubilized in isooctane. The photolability of the aqueous protein when subjected to irradiation at 280 nm is suppressed in the isooctane-soluble complex. 5. Phosphatidylserine brings about a rapid (t 1/2 is about 150 ms at a lipid concentration of 0.75 mM) dissociation of the linear aggregates formed between the enzyme oligomers. 6. A model of the enzyme-lipid-membrane complex, consistent with these results, is proposed. It is suggested that the enzyme is an allotopic protein and that the dissociation of the enzyme in vitro may involve binding sites on the protein which are designed for interaction with the cardiolipin of the inner mitochondrial membrane, when the enzyme is in the mitochondrial matrix.


Purification and properties of pyrazon dioxygenase from pyrazon-degrading bacteria. Chromatography on DEAE-cellulose and gel filtration on Sephadex revealed that pyrazon dioxygenase from pyrazon-degrading bacteria consists of three different enzyme components. No component alone oxidizes the phenyl moiety of pyrazon, only when the three components are combined can oxidation be detected. Following electron paramagnetic resonance and ultraviolet measurements the protein nature of the three components was determined: component A1 (molecular weight about 180000,red-brown in colour) is an iron-sulphur protein. The existence of approximately two moles of iron and two moles of inorganic sulphur per mole of protein was demonstrated. This enzyme component was purified to homogeneity in disc electrophoresis. Component A2 is a yellow protein of a molecular weight of about 67000. FAD was shown to be the prosthetic group of this protein. Component B (molecular weight about 12000, brown in colour) is a protein of the ferredoxin type, which was purified to homogeneity, as demonstrated by disc electrophoresis. A hypothetical scheme for the cooperation of the three components is proposed: component A2 accepts as cosubstrate NADH and functions as a ferredoxin reductase. The ferredoxin, component B, has the function of an electron carrier. The conversion of the substrates is effected by component A1, the terminal dioxygenase.


Antibodies and autoantibodies of glycogen phosphorylase b: inactivation of pig and rabbit enzymes. Pig skeletal muscle glycogen phosphorylase b was purified using ammonium sulfate fractionation, DEAE-Sephadex A-50 and Sephadex G-200 column chromatography. The purified enzyme was used to immunize rabbits in the presence or in the absence of complete Freund adjuvant. Antibodies against pig phosphorylase in pure form were isolated from rabbit antisera using insoluble immunoadsorbents of pig phosphorylase. Autoantibodies against the rabbit enzyme were obtained from the same antisera using insoluble immunoadsorbents of rabbit phosphorylase. Complete inactivation of pig phosphorylase was accomplished by an antibody/enzyme molar ratio equal to 4 and autoantibody/enzyme molar ratio equal to 130. Complete inactivation of rabbit phosphorylase was accomplished by an antibody/enzyme molar ratio equal to 250 and autoantibody/enzyme molar ratio equal to 160. Passive haemagglutination technique gave positive results with minimum amounts of 0.02 microng/ml and 0.8 microng/ml for pig and rabbit phosphorylase respectively. Kinetic experiments have shown that antibodies and autoantibodies act as noncompetitive inhibitors of both enzymes with respect to AMP and glucose 1-phosphate but exhibit a mixed type  of inhibition with respect to glycogen. When glycogen hydrolysates were used as substrate in place of intact glycogen molecules a pronounced decrease in the inhibitory capacity of antienzyme on the enzyme was demonstrated.


Cathepsin L. A new proteinase from rat-liver lysosomes. 1. Cathepsin L was purified from rat liver lysosomes by cell fractionation, osmotic disruption of the lysosomes in the lysosomal mitochondrial pellet, gel filtration of the lysosomal extract and chromatography on CM-Sephadex. 2. Cathepsin L is a thiol proteinase and exists in several multiple forms visible on the disc electropherogram. By polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate its molecular weight was found to be 23000-24000. The isoelectric points of the multiple forms of cathepsin L extended from pH 5.8-6.1 ascertained by analytical isoelectric focusing. 3. Using various protein substrates, cathepsin L was found to be the most active endopeptidase from rat liver lysosomes acting at pH 6-7. In contrast to cathepsin B1, its capability of hydrolyzing N-substituted derivatives of arginine is low and it does not split esters. 4. Greatest activity is obtained close to pH 5.0 with 70-90% of maximal activity at pH 4.0 and pH 6.0 and 30-40% at pH 7.0. 5. The enzyme is strongly inhibited by leupeptin and the chloromethyl ketone of tosyl-lysine. Leupeptin acts as a pseudo-irreversible inhibitor. 6. The enzyme is stable for several months at slightly acid pH values in the presence of thiol compounds in a deep-frozen state.


Anabolic ornithine carbamolytransferase of Pseudomonas. The bases of its functional specialization. The anabolic ornithine carbamoyltransferase of Pseudomonas appears to be extremely specialized. Unlike the other carbamoyltransferases studied, this enzyme catalyzes the phosphorolytic cleavage of citrulline with a very poor efficiency. The main goal of this paper is to understand what, in the catalytic process, causes this directed functional specialization. On the basis of kinetic data and thermodynamic properties of the reaction, it appears that the reaction mechanism is the same as for ornithine carbamoyltransferases from other sources, that is, of the sequential ordered type, where carbamoylphosphate is the first substrate to be bound and phosphate the last product to be released. In addition to this, and here lies the difference with other ornithine carbamoyltransferases, the anabolic transferase of Pseudomonas forms a binary dead-end complex with citrulline, leading to inefficient binding of phosphate and citrulline to the enzyme. Therefore the phosphorolytic cleavage of citrulline is equally inefficient. It should be mentioned that the affinity of the enzyme for citrulline at its catalytic site is low as compared to other transferases.


The kinetics of reoxidation of reduced benzylamine oxidase. 1. The mechanism of reoxidation of reduced benzylamine oxidase has been investigated at different pH between 6 and 10 by steady-state and transient-state kinetic methods. 2. The reoxidation process involves minimally a second-order interaction between reduced enzyme and oxygen leading to the formation of a spectrally modified enzyme intermediate, and a subsequent first-order step converting this intermediate into free enzyme. The variation with pH of rate constants according to such a reaction scheme is reported. 3. Under aerobic conditions the oxygen-independent reaction represents the main rate-limiting step in the catalytic process at alkaline pH. At neutral or acid pH the interaction between reduced enzyme and oxygen becomes mainly rate-limiting, indicating that the concentration of oxygen may be a critical factor controlling enzyme activity under physiological conditions. 4. The spectrally modified intermediate formed during the reoxidation process exhibits a difference-absorption band centered around 290 nm in comparison to free enzyme, and an additional difference-absorption band at 470 nm in comparison to reduced enzyme. These data indicate that formation of the intermediate, besides leading to a reappearance of the 470-nm absorption band disappearing on reduction of the enzyme, results in a spectral perturbation of one or several aromatic amino-acid residues in the protein. This perturbation could possibly reflect a conformational change of the enzymes.


Studies of glutamate dehydrogenase. Methionine-169: the preferentially carboxymethylated residue. In phosphate buffer at pH 7.0, 5,5'-dithio-bis(2-nitrobenzoic acid), N-ethylmaleimide or iodoacetamide do not alter the activity of beef liver glutamate dehydrogenase. Iodoacetate, however, inactivities the enzyme irreversibility by alkylation. Combined addition of the coenzyme NADH and the substrate 2-oxoglutarate or the effector GTP protects against this inactivation. The alkylation reaction is independent of pH between pH 6-9 indicating that amino, imidazole or phenolic groups are probably not involved in this reaction. Titration of the thiol groups, after denaturation of the enzyme, revealed the loss of approximately one group per polypeptide chain. However, this is not due to the exclusive alkylation of a cysteine residue, since alkylation with iodo-[2-14C]acetic acid also labels a methionine residue. 50% of the label is incorporated into methionine-169 and only 7% into cysteine-115, the remaining radioactivity is distributed in minor quantities (4%) in several unidentified residues. A probable cause of the erroneous thiol groups titration is discussed.


Structural studies on periodate-oxidized chicken ovomucoid. Spectral behaviour of the tyrosine residues. About 50% of the carbohydrate moiety of ovomucoid was destroyed by periodate oxidation. The oxidation was carried out for 6 h or 24 h. The data obtained showed that in the carbohydrate chain 2-5 glucosamines and 1-2 neutral sugar residues were decomposed with the consumption of 16 mol and 29 mol of periodate respectively. Periodic oxidation slightly changed the inhibitory activity of the ovomucoid, but altered its spectral properties. An increase of the absorption maximum at 278 nm was noted, as well as a tendency for normalization of phenolic ionization and an increase of the relative fluorescence. The reactivity of tyrosine residues towards tetranitromethane is also changed. It was suggested that even in native ovomucoid the tyrosines could be regarded as 'dissolved' in the 'carbohydrate solvent'. This contact could be achieved by the hydrogen bonds in the formation of which the NHCOCH3 groups of the glucosamine residues play an essential role. Peroxidate oxidation seems to lead to an alteration of the nature of the 'sugar solvent' and disturbs the conformation of the sugar chain.


The purification and properties of NADP-dependent isocitrate dehydrogenase from ox-heart mitochondria. The purification of NADP-linked isocitrate dehydrogenase from ox heart mitochondria is described. The molecular weight from gel filtration, sedimentation equilibrium and gel electrophoresis is 90000+/-4000, and there are two subunits in the molecule each of which binds NADPH with enhancement of the coenzyme fluorescence. The amino-acid composition is reported, and the absorption coefficient, A1/280%, estimated from dry weight measurements is 11.8 cm-1.


Glycosaminoglycans and glycoproteins in metachromatic leucodystrophy. A study is reported of the brain glycoproteins and the glycosaminoglycans in a case of infantile metachromatic leucodystrophy, associated with a ponto-cerebellar systemic atrophy, verified by autopsy. The glycosaminoglycan concentrations in the grey matter were within the normal range, while in the white matter they were slightly increased. The glycopeptides from the soluble and insoluble glycoproteins of white matter were altered not only in their quantity but also in their composition. In the grey matter a decrease of all sugars except the NaNa was detected in the insoluble fraction while in the soluble no change was detected, except for a threefold increase of fucose. The relationship was discussed of soluble to insoluble glycoproteins, and a mechanism for their derangement in MLD was suggested.


Prolongation of rat renal allograft survival by cyclophosphamide and intravenous donor-specific antigens. Specific immunological hyporeactivity to Ag-B incompatible rat renal allografts was achieved after pretreatment of the recipients with donor strain platelets or spleen cells and cyclophosphamide (CY). The longest survival times were observed in animals pretreated with a single 75 mg/kg dose of CY together with 2 X 10(10) donor strain platelets or 2.5 X 10(9) spleen cells intravenously, 2 weeks prior to kidney transplant (median survival time, 71 and 47 days, respectively, compared to 12 days in untreated rats). CY or antigen given alone were ineffective. Anti-donor antibody activiy was routinely detectable in graft-bearing animals. Cell-mediated anti-donor immunity, although impaired, was still present in long-term survivors. These findings suggest that preservation of graft function and prolonged survival in antigen-CY-pretreated animals may be abetted by a combination of mechanisms including antigen-induced immunological enhancement, and deletion by CY of potentially reactive lymphoid cell clones. The use of CY in conjunction with donor antigen pretreatment may provide an additional increment of specific immunosuppression in clinical organ transplantation.


Stress and cardiovascular disease.

Stress and cardiovascular disease. Currently the role of stress as a potential factor in cardiovascular disease is receiving considerable attention. Accordingly, it is important to know how it is defined and identified, how it affects the cardiovascular system and how it can be managed with the knowledge presently available. An overview of the present body of knowledge on the relationship between stress and cardiovascular disease is presented for the reader's consideration.


Alterations of lipid metabolism in healthy volunteers during long-term ethanol intake. Nine young, healthy male volunteers were given ethanol (75 g/day) for 5 weeks. The ethanol was divided into five daily doses and taken so that blood ethanol levels never exceeded 0.04% (w/v). During the latter part of the ethanol intake period, there was a significant, transient increase of plasma triglyceride (TG) concentrations followed by reduction to normal levels. A three-fold increase of lipoprotein lipase activity (LLA) occurred in biopsy specimens of adipose tissue. An increase of alpha-lipoprotein concentrations, which correlated significantly with the decrease in plasma TG levels and the increase in adipose LLA, was also observed during the ethanol intake period. No changes were observed in plasma cholesterol and beta-lipoprotein levels. A transient, three-fold increase of TG concentrations occurred in liver biopsy specimens. Ultrastructural and cytochemical examinations of the biopsy specimens showed hyperplasia of the smooth endoplasmic reticulum, and increased canallicular activity of gamma-glutamyl transferase (gamma-GT) activity in most subjects towards the end of and after the ethanol intake period. Serum gamma-GT levels also increased significantly.


Plasma concentrations, bioavailability and dissolution of chlorpropamide. The bioavailability of chlorpropamide from two new formulations (Melitase tablets) has been compared to that from a reference formulation which is currently in clinical use as a hypoglycaemic agent. In both rate and extent of bioavailability, all three formulations may be considered equivalent, providing allowances are made for differences in drug content. With 95% confidence, the mean bioavailability of chlorpropamide from the new formulations was within about 16% of the mean from the reference formulaion, and formulation-related differences were not statistically significant. Although all three formulations were shown to have similar dissolution  profiles, dissolution of chlorpropamide was pH-dependent in vitro. Dissolution was almost complete during 30 min at pH 7.2, but only 40%-60% had dissolved during 90 min at pH 2.0. A peak mean concentration of 22.7 mug/ml was reached 3 h after administration of 2 x 100 mg tablets of the new formulation and peak mean concentrations of 26.8 mug/ml and 27.4 mug/ml were reached 3 h and 4 hours after administration of one 250 mg tablet of the new formulation and one 250 mg tablet of the reference formulation respectively. Formulation-related differences of mean plasma concentrations (after scaling for equal doses of 250mg) were not significant and each formulation provided similar plasma concentrations at corresponding times after administration. Statistically significant subject-related differences in all the parameters of bioavailability were shown by analyses of variance.


Tachyphylaxis to beta-adrenoceptor agonists in guinea pig airway smooth muscle in vivo and in vitro. Beta-Adrenoceptor tachyphylaxis was induced by incubating spirally cut guinea pig tracheas with isoproterenol (2.4 x 10(-7) M) for 20 min. This incubation reduced the relaxant effects of catecholamines but not of dibutyryl cyclic AMP, theophylline or sodium nitrite. Tracheas incubated with norepinephrine, phosphodiesterase inhibitors or cyclic nucleotides became tachyphylactic to isoproterenol. Pretreatment with indomethacin prevented induction of tachyphylaxis. Incubation with adenosine, methoxamine or sodium nitrite did not induce beta-adrenoceptor tachyphylaxis. When we gave isoproterenol intramuscularly to guinea pigs, airway sensitivity to aerosolized histamine was unchanged but the toxicity of parenterally administered histamine was increased. A prolonged treatment with isoproterenol reduced airway sensitivity to histamine aerosols; this reduced sensitivity was reversed by indomethacin. Thus, beta-adrenoceptor tachyphylaxis may not explain increased toxicity of parenteral histamine after isoproterenol treatment. Elevated levels of cyclic AMP and an increased synthesis of prostaglandins may result in diminished response to beta-receptor stimulation.


Effect of neurohumoral modulators on the morphine-induced hyperthermia in non-tolerant rats. Wistar rats from one supplier have been shown to exhibit the atypical body temperature responses to morphine. In contrast to commonly used rats, in which morphine induced dose-dependent changes in body temperature, the initial administration of morphine (5, 10, 20, and 40 mg/kg, s.c.) to rats of this particular strain produced a consistent and prominent hyperthermia. This hyperthermia is mediated via an action on the typical morphine receptors since it was completely prevented by a specific narcotic antagonist, naloxone. Effects of neurohumoral modulators on the hyperthermia have been investigated in these rats. Pretreatment of animals with p-chlorophenylalanine, alpha-methyltyrosine, phenoxybenzamine or propranolol did not alter the hyperthermia. In contrast, the s.c. administration of 1 mg/kg of either tertiary or quaternary anticholinergic drug such as scopolamine, atrophine, methscopolamine and methylatropine significantly inhibited the hyperthermia. These results suggest that morphine causes  hyperthermia in some strains of rats by a cholinergic mechanism and the involvement of an adrenergic or serotonergic mechanism in this case appears unlikely.


Induction of tolerance and withdrawal in rats receiving morphine in the spinal subarachnoid space. Rats implanted with chronic catheters in the spinal subarachnoid space were given twice daily injections for 7 days of morphine sulfate, either intrathecally into the lumbar subarachnoid space (15 or 50 microng) or i.p. (20 mg/kg). The development of tolerance, as manifested in a reduction of the analgetic efficacy of these injections on the hot plate and tail flick, occurred in a dose dependent fashion over a period of 7 days. At this time, injections of i.p. morphine into animals which had received spinal morphine and vice versa revealed the existence of a two way cross tolerance between spinal and systemically administered morphine. Injection of naloxone into the spinal cord of animals exposed to i.p. morphine or conversely, i.p. naloxone in animals tolerant to intrathecal morphine, yielded a hyperreflexia and extreme sensitivity to handling. Other signs commonly observed in percipitated withdrawal, however, such as wet shakes and weight loss, were not observed.


Long-loop reflexes in the tranquilized monkey. EMG responses to sudden displacement of the forelimb were studied in Cebus monkeys tranquilized with Atravet, a phenothiazine tranquilizer. The monkey's forearm was strapped firmly to a manipulandum handle. A torque motor attached at the pivot point of the handle, under servo control, provided reproducible limb displacements. In response to a sudden maintained displacement three periods of EMG activation in biceps muscle occurred with peak latencies of approximately 25, 45 and 85 msec. These correspond to the latencies of the M1, M2 and M3 responses in the alert animal. Similar responses were observed in 'naive' animals which had not previously been used in experimentation. All three responses increased in magnitude with increasing background activity and all appeared to be associated with suppression of EMG activity in the antagonist muscle. M1 and M2 responses were position dependent, M1 being greater in extension than in flexion and M2 the opposite. The position-dependence of the M2 response was produced by a depression of activity following the M1. This depression of activity lasted up to 30 msec following M1 and was directly dependent on the M1 magnitude.


[Effect of dimedrol, diazoline, diprazin and ethysine on formation of the primary immune response]. Tests were conducted on mice of the CBA line, immunized through a single intraperitoneal administration of 0.5 ml of a 5% sheep erythrocytes suspension. Antihistaminic agents were introduced by the intraperitoneal route in a dose of 5 mg/kg, first at one time and then repeatedly at various intervals with respect to the initial immunization benadryl and diazoline, when administered in a single dose to mice 24 hours after immunization activated the primary immune response to sheep erythrocytes, whereas phenergan and ethisine manifested merely a tendency toward activation of the immunogenesis. The drugs under study failed to have any significant effect on the titres of serumal hemolysines and hemaglutinines.


[Effect of cholinergic substances on brain potentials induced by tooth pulp stimulation]. Introduction of the M-cholinomimetic -- arecoline into the ventricle of the rabbit's brain decreases and of the M-cholinolytic -- atropine -- increases the amplitude of primary responses in the sensomotor region of the cortex, arising out of stimulating the dental pulp. The N-cholinomimetic -- lobeline potentiates the evoked response of the brain cortex, whereas the N-cholinolytic -- beta-ethylene-splasmolytin -- depresses it. It is assumed that the region of the drugs action application are thalamic nuclei.


[Phencarol--an antihistaminic preparation from the quinuclidylcarbinol group]. A high antihistaminic activity of quinuclidyl-3-diphenylcarbinol (phencarol) has been ascertained. In pharmacological experiments phencarol is superior to dimedrol (diphenhydramine), both as concerns its antihistaminic activity and the duration of its action. Unlike dimedrol and diprazine (promethazine) it has no inhibitory effect on the central nervous system. With its clinical use phencarol is effective in treating urticaria, Quincke's edema and allergic skin reactions. The drug is well tolerated by the patients and does not produce any inhibitory or soporific action. Phencarol has an authorized permission to be used for medical purposes.


[Study of the effect of corticosterone on erythrocyte membranes by the spin label method]. Influence of corticosterone on the hare's erythrocyte ghosts is studied by the spin-label method. There exists a correlation between the maximal variation region in the rotational diffusion "frequency" of the spin-labels, covalently linked with the membrane protein, and the hormones concentration range within which the effect of corticosterone on the functions of the erythrocyte membranes manifests itself to the greatest degree (permeability to ions, hemolytic effects, etc).


[Effect of narcotics on impulse conduction in the afferent pathways of visceral nerves]. In tests set up on cats immobilized with pyraxolone and anatruxonium the effect of non-inhalation narcotics on the evoked potentials of the cortex and subcortical structures with visceral and also somatic, acoustic and photo-stimulation was studied. With stimuli of different modality sodium ethaminal was found to inhibit evoked potentials of the cerebral hemispheres, diencephalon and midbrain cortex. Hexabarbital sodium suppressed biopotentials in specific, associative and non- specific brain structures in the event of visceral and somatic stimulation. Viadril enhanced the amplitude of potentials in all types of stimulation Urethan inhibited potentials in specific pathways of visceral and somatic nerves as well as in associative and non-specific brain structures following visceral, somatic and acoustic stimulation, with the amplitude of potentials to photostimulation then increasing.


[Effect of several psychopharmacologic preparations on adaptation to stressful conditions]. Experiments staged on rats demonstrated that the formation of pathological states caused by stress and accompained by the development of ulcerative lesion of the gastric mucosa are associated with the degree of the catecholamines level drop in the mesencephalon and hypothalamus. The application of seduxen and also of combinations consisting of L-DOPA with seduxen, or with an L-adrenoblocking agent-pyroxan tends to reduce the frequency of developing ulcerative lesions of the stomach. The protective effect produced by the combination of L-DOPA with an L-adrenoblocking agent--pyroxan is barred by an additional administration of an beta-adrenoblocking agent--inderal.


The photochemical cycle of bacteriorhodopsin. The reaction cycle of bacteriorhodopsin in the purple membrane isolated from Halobacterium halobium has been studied by optical absorption spectroscopy using low-temperature and flash kinetic techniques. After absorption of light, bacteriohodopsin passes through at least five distinct intermediates. The temperature and pH dependence of the absorbance changes suggests that branch points and/or reversible steps exist in this cycle. Flash spectroscopy in the presence of a pH-indicating dye shows that the transient release of a proton accompanies the photoreaction cycle. The proton release occurs from the exterior and the uptake is on the cytoplasmic side of the membrane, as required by the function of bacteriorhodopsin as a light-driven proton pump. Proton translocating steps connecting release and uptake are indicated by deuterium isotope effects on the kinetics of the cycle. The rapid decay of a light-induced linear dichroism shows that a chromophore orientation change occurs during the reaction cycle.


Photochemical and chemical studies on the chromophore of bacteriorhodopsin. The chromophore (purple complex) of bacteriorhodopsin is reduced by sodium borohydride upon illumination to RPhv with a three-peaked absorption band at 360 nm. Treatment of this reduction product with ultraviolet light or acid yields a modified product from which retro-retinyllysine can be obtained by alkaline hydrolysis. No reduction of the 412 nm complex was found. Under specific conditions the purple complex equilibrates with a photochemically active 460 nm form that can be reduced by borohydride in the dark. This reduction product RP460 behaves idential to RPHV. Reconstitution of the purple complex from chromophore-free membrane (apomembrane) and retinal occurs via intermediates. The first (lambdamax 400nm) shows a three-peaked absorption band and is reduced to RP400 without a change of the three-peaked absorption (lambdamax 360 nm). The same product is obtained from apomembrane and retinol. Detergents shift the absorption band to 330 nm in all cases. From the experiments described no participation of retro-retinal structures during the photochemical cycle can be concluded but stereospecific interaction of the retinal moiety with the protein resulting in a specific retinal conformation os omdocated by the spectral changes observed.


Proton translocation by ATPase and bacteriorhodopsin. Stable membrane proteins and lipids are convenient to study biomembranes. Two stable proton translocating proteins were purified and reconstituted into vesicles capable of proton translocation. One was a thermostable ATPase (TF0-F1) of thermophilic bacterium PS3 and the other was rhodopsin of Halobacterium halobium. TF0-F1 was composed of a proton pump moiety (TF1) and a proton channel moiety (TF0). TF1 was the first membrane ATPase which was crystallized and reconstituted from its five polypeptides. Like TF0 and TF1, the rhodopsin in purple membrane was highly stable against dissociating agents, acids and alkali. Phospholipids of these biomembranes were also stable and contained no unsaturated fatty acyl groups. The molecular species of the phospholipids of PS3 were determined by mass chromatography. Measurements were made of the difference in electrochemical potential of protons (deltamicronH+) across the membrane of the reconstituted vesicles. The deltamicronH+ attained was 312 mV in TF0-F1 vesciles and was 230 mV in the rhodopsin vesicles. To conclude that electron transport components are not necessary for ATP synthesis in energy yielding biomembranes, two experiments were performed: The ATP synthesis was observed i) on acid-base treatment of TF0-F1 vesicles, and ii) on illumination of the rhodopsin-TF0-F1 vesicles.


Light-dependent cation gradients and electrical potential in Halobacterium halobium cell envelope vesicles. Vesicles can be prepared from Halobacterium halobium cell envelopes, which contain properly oriented bacteriorhodopsin and which extrude H+ during illumination. The pH difference that is generated across the membranes is accompanied by an electrical potential of 90-100 mV (interior negative) and the movements of other cations. Among these is the efflux of Na+, which proceeds against its electrochemical potential. The relationship between the size and direction of the light-induced pH gradient and the rate of depletion of Na+ from the vesicles, as well as other evidence, suggest that the active Na+-extrusion is facilitated by a membrane component that exchanges H+ for Na+ with a stoichiometry greater than 1. The gradients of H+ and Na+ are thus coupled to one another. The Na+-gradient (Na+out greater than Na+in), which arises during illumination, plays a major role in energizing the active transport of amino acids.


Light-activated amino acid transport in Halobacterium halobium envelope vesicles. Vesicles prepared from Halobacterium halobium cell envelopes accumulate amino acids in response to light-induced electrical and chemical gradients. Nineteen of 20 commonly occurring amino acids have been shown to be actively accumulated by these vesicles in response to illumination or in response to an artificially created Na-gradient. Sodium-activated amino acid transport for 18 of these amino acids has been shown to occur in direct response to the protonmotive force generated. Glutamate is transported only in response to a sodium gradient. Michaelis constants for the uptake of these amino acids are close or identical whether the amino acids are accumulated in response to a sodium gradient or a protonmotive force (i.e., electrical gradient). On the basis of shared common carriers the transport systems can be divided into eight classes, each responsible for the transport of one or several amino acids, i.e., arginine, lysine, histidine; asparagine, glutamine; alanine, glycine, threonine, serine; leucine, valine, isoleucine, methionine; phenylalanine, tyrosine, tryptophan; aspartate; glutamate; proline. Available evidence suggests that these carriers are symmetrical in that amino acids can be transported equally well in both directions across the vesicle membranes. A tentative working model to account for these observations is presented.


Light energy conservation processes in Halobacterium halobium cells. In Halobacterium halobium, proton pumping driven by light or by respiration generates an electrochemical potential difference across the membrane. Energy storage in this form is only transient. Cellular energy transducers competing with proton leaks stabilize this free energy as high energy phosphate bonds, electrochemical potential of other ions, and chemical potential of amino acids and possibly other chemical species. The pH changes induced by light or by respiration in cell suspensions are complicated by proton flows associated with the functioning of the cellular energy transducers. Dominant is the proton inflow coupled to the synthesis of ATP, which has been kinetically resolved. A proton-per-ATP ratio of about 3 is calculated from simultaneous measurements of photophosphorylation and the proton inflow. This value is compatible with the chemiosmotic coupling hypothesis. The time course of the light-induced changes in membrane potential indicates that light-driven pumping increases a dark preexisting potential of about 130 mV only by a small amount (20-30 mV). The complex kinetic features of the membrane potential changes do not closely follow those of the pH changes, indicating that flows of ions other than protons are involved. A qualitative model consistent with the available data is presented. A salient feature of this model is a sudden relaxation of the protonmotive force by a proton inflow through the ATPase when the preexisting protonmotive force is increased by light or respiration and reaches a critical value. The trigger could be either the proton-motive force, the pH gradient, or possibly the internal pH.


Genetic counselling and prenatal diagnosis for chromosome anomalies. Use of study of spontaneous abortions. Genetic counselling for chromosome abnormalities must take into account the fact that such anomalies may develop as an error in oogenesis or spermatogenesis or may be transmitted from parents. Since many chromosomally abnormal embryos are spontaneously aborted, parents with repeated unfavorable outcomes of pregnancy or difficulty in conceiving should be studied by karyotype. The relationship between chromosomal abnormalities and various environmental or biologic factors are discussed. The outcome of a previous pregnancy may indicate the necessity of subsequent fetal evaluation even with chromosomally normal parents.


Prenatal diagnosis of chromosome abnormalities. It is possible to define a group of prospective parents who are at risk of producing offspring with disability due to chromosomal abnormalities. Such individuals should have evaluation of the 15 or 16 week fetus by karyotype of amniotic fluid cells. The techniques and problems of such diagnostic procedures are discussed.


Prenatal diagnosis of inborn errors of metabolism. More than 50 inborn errors of metabolism can be diagnosed in the fetus. This is accomplished by evaluation of constituents of the cell free amniotic fluid or of cultured or uncultured amniotic fluid cells. Only those syndromes with specific abnormal contributions to amniotic fluid can be so diagnosed. Currently diagnosable diseases and their assay methods are reviewed. Such assay methods must be able to differentiate the unaffected heterozygote from the affected homozygote.


Diagnostic radiology of fetal abnormalities. Fetal abnormalities involving the skeletal system, central nervous system or soft tissue may be diagnosed by radiologic methods, as may fetal death. Occasionally standard X-ray is used, but often invasive techniques are required, and should be limited to those patients with valid indications. X-rays of specific fetal anomalies are presented.


New and future techniques in prenatal diagnosis. Fetal evaluation may be accomplished in the future using techniques experimental at this time. These include direct visualization of the fetus by endoscopy, biopsy of fetal tissues, and indirect techniques accomplished in the laboratory. It is hoped that in the future technology may provide us with means to diagnose the abnormal fetus more accurately and rapidly than currently. Possible methods are discussed.


Disappearance of para-amino-hippurate from amniotic fluid. Eight pregnant women and three pregnant sheep received 400 mg of para-amino-hippurate (PAH) intraaminotically. Serial samples of amniotic fluid and maternal blood were obtained. In sheep samples of fetal blood were also withdrawn. PAH appeared in maternal plasma in all the cases. In all pregnant women PAH disappeared slowly from amniotic fluid (50% in 4 hours). In one ewe the study was performed as in humans and showed the same pattern of disappearance. In the other two, fetal urine was drained outside the amniotic fluid and PAH disappeared from it at a much faster rate (90% in 4 hours). PAH concentration in fetal urine was 100 times higher than in fetal plasma. Our findings in pregnant women seem to suggest that PAH disappears from the amniotic sac by a diffusion mechanism. On the other hand the results found in sheep also suggest that the fetus may have an active role in PAH concentration in amniotic fluid, eliminating part of the substance into maternal blood across the placenta but returning a major portion to the amniotic fluid with fetal urine.


Antepartum fetal arrythmia. A case report. On a number of occasions fetal heart arrythmias have been detected during the course of routine antepartum patient examination. A case is presented to illustrate the difficulty of making a definitive diagnosis of the nature of the arrythmia, and its significance in relation to fetal status. Investigation included stressed and nonstressed FHR monitoring, but a definitive diagnosis could not be made during the antepartum period and the patient could, therefore, not be assured of the essentially benign nature of the arrythmia until shortly after the birth of her child. The electrocardiogram obtained from external fetal electrocardiographic monitoring is not sufficiently refined to aid in the antepartum diagnosis of fetal cardiac arrythmias.


The estimation of gestational age by amniotic fluid analysis and its relationship to fetal growth retardation. The relationship between three amniotic fluid parameters (fat staining cells, anucleate cells and creatinine) and gestational age has been studied in 166 samples from 107 patients of known gestational age. Individually there is an unacceptable degree of inaccuracy. However, when the three parameters are combined in a multilinear regression, an equation for gestational age is produced that is reliable to with  +/-14 days in over 90% of cases. This held true when the equation was applied to 60 samples from 45 patients with uncertain dates and related to the paediatric estimate of gestational age. The accuracy was unaltered by intrauterine growth retardation.


Repeated extra-amniotic administration of prostaglandin F2alpha for midtrimester abortion. Midtrimester abortion was induced in 94 of 100 patients at 16 to 24 weeks' gestation by the extra-amniotic administration of 1170 microng of prostaglandin F2alpha (PGF2alpha) every 10 minutes. The number of prostaglandin doses varied from 16 to 24 depending on the patient's response to the prostaglandin. The median abortion time was 10.0 hours, and 82.0% of the patients aborted within 24 hours. Overall, 68.0% of the patients failed to expel the placenta within one hour of abortion of the fetus. Vomiting and diarrhea occurred among 42.0 and 17.0% of the patients, respectively. Compared with the intra-amniotic administration of a single 50 mg dose of PGF2alpha, the extra-amniotic procedure was associated with similar side effect rates, a higher rate of incomplete abortion, and a significantly shorter abortion time.


Pregnancies in the presence of copper intra-uterine devices. Hysterosalpingographic studies. Two copper-bearing intrauterine devices--the Cu-7 and Cu-T-200--were investigated to determine why contraceptive failures occur with these devices. For this investigation, 155 multiparous women were followed up for at least 24 months after IUD insertion (T-Cu-200 in 75 cases and the Cu-7 in 80 cases). Three women using the T-Cu-200 and 5 women using the Cu-7 became pregnant during the first year. Hysterosalpingographic findings are presented for 8 of these cases and for 2 additional cases (one with each device) who became pregnant during the second year of follow-up. These indicate that uterine abnormalities and disproportions between the uterine cavity's shape and size and that of the device are the primary reasons for device displacement and partial fundal coverage leading to contraceptive failures.


Estrogen-progesterone withdrawal bleeding in diagnosis of early pregnancy. This study was designed to evaluate the efficacy of an exogenous estrogen-progesterone preparation for inducing "withdrawal bleeding" in non-pregnant women, a procedure used in some areas for the diagnosis of early pregnancy. Three hundred patients not desiring to be pregnant and with no signs of pregnancy other than menses delayed by 14 days or less were randomly assigned to treatment or control groups. The treatment group received an intramuscular injection of 50 mg progesterone and 3 mg oestradiol benzoate in oil. The control group received no hormonal injection. There was no significant difference between the two groups in the incidence of uterine bleeding within 7 days. Thus, the hormonal preparation was ineffective in inducing "withdrawal bleeding". Its use as a diagnostic test for pregnancy is not recommended.


Carcinoma in situ of the Fallopian tube associated with cervical carcinoma. Case report. A case of carcinoma in situ of the fallopian tube in a 55-year-old woman, associated to cervical carcinoma is described. The necessary criteria for the diagnosis of pre-invasive carcinoma of the fallopian tube, among which the number of mitoses seems to be of great importance, are discussed. This previously unreported association of carcinoma in situ of the fallopian tube, with carcinoma of the uterine cervix, emphasizes the notion of the multicentric neoplastic possibilities of the müllerian tract derivatives.


A transcutaneous PO2 electrode for continuous monitoring of fetal state during delivery. A transcutaneous PO2 electrode has been designed and constructed for the continuous measurement of fetal oxygen tension during labor and delivery. The electrode, tested in vitro, has a low oxygen consumption, a short response time, 10 sec, and a sufficient stability, 5% over 3 hr. The in vivo performance of the electrode, tested in neonates, shows a good correlation between transcutaneous PO2 and PO2 measured in samples of arterial blood. The continous registration of fetal tcPO2 during labor and delivery is exemplified.


A new technique and instruments for delivery of the Fallopian tube in female sterilization. This report describes an instrument designed for locating and delivering the Fallopian tube through an incision of less than 2 cm. Since no gas insufflation, fibre optic illumination, or electricity is required for successful use of this instrument, it is well suited for performing outpatient tubal sterilizations in areas where other methods, such as laparoscopic procedures, are not feasible.


The effect of indomethacin in labour at term. The effect of indomethacin as an antagonist to prostaglandin was evaluated on a series of 16 women during spontaneous labour in the ninth month of pregnancy. The evaluation of the effect of indomethacin on the progression of labour was determined by the deviation from the normal curve of Friedman. In 8 cases there was complete cessation of labour from 3 to 192 hours and in 8 additional cases there was protraction of the active phase from 3 to 17 hours. In 2 cases there was no effect. Seven births terminated spontaneously and in 9 cases it was necessary to administer pitocin intravenously to strengthen or renew contractions in order to terminate the pregnancy. There is no correlation between the obstetric state (the degree of cervical dilation, or effacement or rupture of membranes) at the time of indomethacin administration and the uterine response, or the speed of return to the normal curve of Friedman. The effect of indomethacin, as an antagonist to prostaglandin, at the time of labour was studied.


Iron deficiency anaemia in Nigerian pregnant women. Ninety-four Nigerian pregnant women, many of who had mild to moderate anaemia, were investigated for the presence of iron deficiency. Using the bone marrow as the diagnostic index, 90% of these patients were found to be iron deficient. Iron deficiency is thus common among our expectant mothers. They should, therefore, be given iron supplement in addition to the present practice of folic acid and antimalarials, throughout pregnancy and the puerperium.


A suggested set of working definitions and criteria applicable to induction of labour. A list of prerequisites, essential items of information and definitions applying to studies on induction of labour is presented. It is suggested that their use would permit proper analysis and comparison of results obtained by different investigators.


Endometrial control: a comparative study of three oral contraceptives. Shortly after completing an OC cycle the relative hormonal deficit caused by discontinuing the hormones results in withdrawal bleeding. Similarly, if one or more oral contraceptive tablets are missed during a cycle, the resulting deficiency may cause breakthrough bleeding. A lower incidence of breakthrough bleeding on the day after one oral contraceptive tablet is missed and later onset of withdrawal bleeding indicate that a particular OC provides better endometrial control. These two measurements are used as indices to compare the endometrial control provided by Ovral, Norinyl 1/50, and Nortestrin 1 in a study of 480 women at the Planned Parenthood Clinic in Seattle during 1974 and 1975. An analysis of the data shows that the normal chance of breakthrough bleeding on a particular day when one oral contraceptive tablet is not missed on the previous day is 0.017, 0.022, and 0.022 for Ovral, Norinyl, Norlestrin users, but the chance on the day after a contraceptive tablet is missed increased by 13 to 16 times for each of the three OCs. Ovral provides more sustained endometrial control than Norinyl and Norlestrin, however, as indicated by its lower probability of breakthrough bleeding on the day a contraceptive tablet is missed and by its later onset of withdrawal bleeding.


Minilaparotomy for female sterilization: a feasibility study of a new technique. Female sterilization via a minilaparotomy incision was performed on 30 patients at Al-Azhar University Teaching Hospital in Cairo, Egypt. A uterine sound fed through a Foley catheter was used as a uterine elevator. A 2 cm suprapubic transverse laparotomy was performed and sterilization was accomplished by fimbriectomy. Twenty patients received general anesthesia and 10 patients initially received local anesthesia. Surgical time ranged from 7 to 15 min, and the average duration of hospitalization was 8 hours. Blood loss was small, and no patient was transfused. One patient had a wound infection. This pilot study indicates that this procedure is relatively simple, safe and inexpensive.


Midtrimester abortion: a comparison of intraamniotic prostaglandin F2alpha and hypertonic saline. The safety and effectiveness of two intraamniotic prostaglandin F2alpha (PGF2alpha) dose schedules (a single 50 mg dose and a repeated 25 mg dose) and intraamniotic hypertonic saline were evaluated in a study where each abortion procedure was randomly assigned to 50 patients. All patients were at 16 to 20 weeks' gestation. Rates of gastrointestinal and other side effects were generally higher for the 50 mg PGF2alpha dose schedule than for the other two procedures. The repeated 25 mg PGF2alpha dose schedule resulted in higher 24-hour (68.0%) and 48-hour (98.0%) cumulative abortion rates than the 50 mg PGF2alpha dose schedule (54.0%, 92.0%) or saline (34.7%, 91.8%). Rates of spontaneous placental expulsion were highest for the repeated 25 mg PGF2alpha dose (74.0%) and lowest for the 50 mg PGF2alpha dose schedule (40.0%).


An assessment of the side effects of switching from one oral contraceptive to another. Side effects associated with three oral contraceptives were evaluated in a study in which women were switched to Norlestrin 1 from either Ovral (64 subjects) or Norinyl 1/50 (26 subjects). In the cycle prior to crossover, breast discomfort was more frequent among Norinyl users than among Ovral users. The prevalence of all other reported side effects was not significantly different for Norinyl and Ovral. The crossover to Norlestrin did not significantly change the numbers of patients reporting side effects. By the end of third Norlestrin cycle, rates of all side effects were similar for women who were switched from either Ovral or Norinyl.


Practicability of ultrasonography for assessing fetal age and weight in early pregnancy. This study was conducted to evaluate (1) the feasibility and accuracy of the B-Scan technique for measuring biparietal diameter (BPD) in utero, (2) the accuracy of the B-Scan technique for predicting fetal age and weight, and (3) parameters of fetal growth in pregnancies of less than 21 weeks's gestation. Forty-one subjects underwent diagnostic ultrasonography before induction of therapeutic abortion with Prostaglandin F2alpha. Following abortion, fetal crown-to-rump length (CRL), crown-to-heel (CHL), BPD, and fetal weight were measured directly. After 13 menstrual weeks' gestation the BPD could be accurately measured by ultrasound. The relationship of BPD to gestational age and fetal weight and the indices of fetal growth as measured by the CRL and CHL were established. It is concluded that ultrasound is a useful adjuvant to clinical assessment of fetal age and weight in patients form 13 to 21 menstrual weeks' gestation.


Laparoscopic sterilization in the puerperium: a review of international experience. The feasibility of using laparoscopic techinques for pueperal sterilization has been demonstrated on three continents. While the operation is not suggested to replace all other methods of pueperal sterilization, it is advantageous in selected situations.


A comparative study of electrocoagulation and tubal rings for tubal occlusion at laparoscopy. The surgical and early postoperative complications and complaints associated with laparoscopic sterilization using electrocoagulation or tubal rings for tubal occlusion were evaluated in a comparative study. Procedures were randomly assigned to subjects (electrocoagulation to 151 subjects and tubal rings to 148 subjects). Difficulties in carrying out the sterillization procedures occurred more frequently when tubal rings were used (6.1%) than when electrocoagulation was used (2.0%). Rates of complications occurring at the time of surgery were similar for the two procedures. However, one patient in the electrocoagulation group had a bowel burn. Postoperative pelvic and abdominal pain were reported by a significantly higher proportion of the tubal ring cases (32.0%) than electrocoagulation cases (19.4%). It appears that although the incidence of surgical difficulties and pain may be somewhat more frequent with the tubal ring, the avoidance of such potentially serious complications as bowel burns may make femal sterilization by the tubal ring method preferable.


Prophylactic antibiotic in elective gynaecological and obstetrical major surgery. A prospective controlled study of 580 patients who underwent non-radical major gynaecological or obstetrical surgery examined the use of prophylactic antibiotics. The test group of patients (290) showed a significantly lower (P=0.01) incidence of the total complications and the non-infective complication rate as against those among controls (290). The duration of hospitalisation was significantly reduced for both vaginal and abdominal surgery in the test group of patients as against those in the controls. There was no evidence of increased incidence of bacterial resistance or superinfection. The authors recommend the use of prophylactic antibiotic on a short term basis (for a total duration of 4 days), commencing preoperatively to achieve a significant reduction in the infective postoperative morbidity and a shorter stay by the patients.


Ossification centers as evidence of fetal lung maturity. Fetal pulmonary maturity based on visualization of one or both fetal knee secondary centers of ossification was assessed in 84 gravidas. Amniotic fluid lecithin/sphingomyelin ration of 2:1 or greater was the criterion of fetal lung maturity. It was concluded that when antenatal radiographs reveal the presence of both centers of ossification, fetal lung maturity can be assured in 100% of the fetuses. When the distal femoral center along is seen, only 60% of the fetuses will have mature lungs.


A comparative study of hysterosalpingography and laparoscopy in the investigation of infertility. A comparison of hysterosalpingography and laparoscopy was carried out on 79 infertile women. Out of these, 60 had abnormal and 19 had normal hysterosalpingograms. Of the 19 patients with normal X-rays, nine (47%) had abnormal findings during laparoscopy. In 62% of patients with abnormal hysterosalpingograms, laparoscopic findings confirmed the radiological diagnosis, however in 38% the findings differed. Though hysterosalpingography is an important part of infertility evaluation, a final and definite diagnosis requires endoscopic evaluation of the pelvis.


Morphologic studies on the non-cancerous cervix in Jamaicans at different ages. A morphologic investigation on 100 cervices removed at autopsy from negro Jamaican women at different ages showed that columnar epithelium was frequently located on the portio vaginalis at all ages. The absence of this phenomenon in some fetal cervices supports the hypothesis of embryologic displacement rather than stimulation by maternal estrogens. Squamous metaplasia was common at all ages, even in the fetus; it showed little relationship to chronic inflammation, but the high frequency in post-pubertal cases (80%) could have some relationship to the high incidence of cervical cancer in Jamaica. Chronic inflammation was not observed in the fetal cervix but occurred in most infants in whom it was attributed to congenital "erosion". However, the cervices of children revealed a marked discrepancy between the presence of "erosion" and the degree and frequency of chronic inflammation (83%). This change is unexplained and requires further study.


A technique for the induction of labor by the extraovular administration of prostaglandin F2alpha. Prostaglandin F2alpha (PG F2alpha) was administered to pregnant women at a dosage of 50 microng or 100 microng by extraovular route around the expected time of childbirth in order to induce labor. (1) PG F2alpha 50 microng or 100 microng dissolved in 10 ml of physiological saline solution was infused by extraovular administration through the cervical canal of the uterus with the use of Nelaton catheter No. 9. The catheter was then immediately withdrawn and the patient allowed to return home. (2) Instilation of 10ml of physiological saline solution was carried out for control purposes. (3) Those who reported labor contractions within 24 hours after the infusion were regarded as cases of excellent response, and those who reported labor contractions more than 24 hours after the infusion, or those who showed dilatation and improvement in effacement of the uterine cervical canal though without the occurrence of labor contractions were deemed to be cases of good response. The overall positive response rate (excellent plus good) was 78.9% for primiparae and 85.7% for multiparae in the group receiving 50 microng and 89.5% for primiparae and 100% for multiparae in the group receiving 100 microng. (4) Labor contraction were induced at a rate of almost 100% in those who showed dilatation of the cervical canal of the uterus to the extent of 3 cm or more and effacement to the extent of 50% or more. (5) It is advisable to perform reinfusion of PG F2alpha or instilation of PG F2alpha as a post-treatment in patients with good responses. (6) The infusion of PG F2alpha is effective for the induction of labor in older primiparae, cases of breech presentation and suspected cases of CPD. (7) There were some cases of enhanced response when PG F2alpha was given as posttreatment.


Plasma estradiol-17 beta as an index of fetoplacental function. Serial plasma unconjugated estradiol-17 beta was measured by a radioimmunoassay method in 26 cases of hypertensive disorder of pregnancy, 13 cases of poor obstetric history and 4 cases of intra-uterine growth retardation. There was considerable overlap of hormonal values between pregnancies, resulting in delivery of normal fetuses and those with dysmature fetuses. Serial estimation of estradiol-17 beta were not found to be useful in the prediction of intra-uterine death, although low values were obtained after fetal death has occurred. Estradiol-17 beta estimation appeared to be an unreliable index of fetoplacental function.


Studies on maternal mortality in Mexico. The sociocultural, psychological, and medical effects of maternal death are described and analyzed for the purpose of appraising the value of the given death as a measure of the quality of medical care. The work of the Maternal Mortality Committee of the Hospital de Gineco-Obstetetricia No. 1 of the Mexican Social Security Institute over its three years of operation and some results on responsiblity for death and its possible predictabilty are presented. The results show that it is necessary to broaden the Committee's functions in a social, professional, and institutional context so that the indicated factors may be objectively assessed and also that the Committee be transformed into a dynamic organism that will contribute to solving the problem. Recommendations for the operation of the Maternal Mortality Committees are given.


Response of fetal acid--base balance to duration of second stage of labour. Data from 4081 vaginal deliveries (no caesarean sections) were analyzed retrospectively (IBM 730/158) with regard to duration of second and "final stage" of labour and fetal outcome (acid-base balance and apgar scores). The average duration of the two periods amounted to 22.8+/-29.6 and 9.9+/-7.8 min. Mean pH (UA) was 7.268+/-0.084 and the acidotic risk (pH less than 7.20, pH less than 7.10) 13.4 and 1.6% respectively. The distribution of the Apgar-scores after 1 min was assessed: 0-3: 1.7%, 4-6: 5.4%, 7-10: 92.9%. The dependance of the time variables from parity was studied. Two samples (N1=1755, N2=1098) of uncomplicated term pregnancies were chosen according to 7 clinical selection criteria differing only in the presence of cord entanglements at birth. The association between the two time variables and parameters of the fetal acid-base balance in cord blood was evaluated using rank correlation- and polynomial regression analysis. Highly significant correlations (tau) were found between the variable time and actual pH in blood of the umbilical artery and vein as well as pCO2, BEECF and HbO2 in the umbilical vein. The association however is not very close and thus clinically not of great importance. The deltapH (UA & UV) pro 60 min of second (and "final") stage of labour was computed and amounted to -0.024 (-0.087) in blood of the umbilical artery and -0.036 (-0.115) in blood of the umbilical vein and -0.017 (-0.062) (UA) and -0.032 (-0.120) units (UV) in the sample with and without apparent cord entanglements at birth respectively. The response of fetal acid-base balance to cord compression during second stage of labour was assesses: The acidotic risk (pH less 7.2) was doubled: 14.5% (7.7%) and AV-differences of all variables were "opened" if cordcoilings were observed. Apgar scores were not significantly different. Moreover, the association between AV-differences of each parameter and the variable time was studied: it became evident that with passage of time AV-difference is "closed" (-0.052 AV DpH/60 min "final stage" of labour, N=1098) indicating time related impairment of placental function. From these observations and data of the literature the conclusion is drawn that second stage of labour should not exceed 45 min in any patient. Furthermore it is concluded that in cases without signs of impending fetal distress it seems to be possible to wait more than 20 (Multipara) or 30 min (Primipara) duration of "final stage" without increased risk of fetal peril measured in terms of acidemia and clinical depression. This is valid only in term pregnancies with the possibility of continuous monitoring of FHR, in cases with normal uterine activity, uneventful course of first stage of labour and cooperative, vigorous patients. The indications for termination of delivery by vaginal operations in cases without impending fetal distress are discussed.


Ectopic pregnancy in southern Iran: a statistical review of 96 cases. A review is presented of 96 cases of ectopic pregnacy over a 7-year period. It is noted that in recent years the incidence of this problem has increased. Inadequately diagnosed and treated pelvic inflammatory disease has been found to be the main predisposing factor (40%). The rate of extra-uterine gestation was 1:336 deliveries. In this study, previous fertility was high, and primigravidae constituted only 10.4% of the total. The effect of previous pelvic operations and pelvic infection is discussed; a statistical review of signs and symptoms is tabulated, and the management of the patient and of concomitant problems is presented.


Pyometra.

Pyometra. Thirteen cases of pyometra without preoperative diagnosis in which vaginal or abdominal surgery was performed, are analysed in the present paper. The histologic examination revealed endocervicitis in all cases, while positive cultures for bacteria were found preoperatively in only eight of the above cases, thus, pyometra was considered to be a symptom of obstruction of the cervical canal.


2,3-diphosphoglycerate, nucleotide phosophate, and organic and inorganic phosphate levels during the early phases of diabetic ketoacidosis. The relation between serum and red blood cell (RBC) inorganic phosphate levels, RBC 2,3-diphosphoglycerate (2,3-DPG) levels, RBC nucleotide phosphate (Pn), and RBC total phosphate (Pt) levels were studied during the early phases of treatment and recovery from diabetic ketoacidosis (DKA). A steady drop in serum inorganic phosphate was found during the first 24 hours of insulin treatment and was most profound at 24 hours. No statistically significant changes (P less than 0.05) were found in red cell inorganic phosphate or nucleotide phosphate levels during the 24-hour study period. The levels of total red cell phosphate were lower in this group of patients than in nonacidotic diabetic subjects and decreased slightly after 24 hours of treatment. The red cell 2,3-DPG levels were low at the initiation of therapy and remained low during the 24-hour study period. Glucose, bicarbonate, lactate, and ketone levels fell in linear patterns with treatment. In view of the current evidence for the effects of low 2,3-DPG on oxygen delivery and the relation of low serum phosphate levels to RBC glycolysis and 2,3-DPG formation, this study reemphasizes the need for phosphate replacement during the early phases of treatment of DKA.


Stimulation of microsomal NADPH oxidation by quinone group-containing anticancer chemicals. Several anticancer chemicals containing a quinone group were found to stimulate the aerobic oxidation of NADPH by liver microsomes. The enzyme responsible for the above reaction was identified as NADPH-cytochrome c reductase (EC 1.6.2.4), one of the microsomal flavoproteins. The fact that a catalytic amount (20 micronM) of these anticancer chemicals was sufficient to oxidize all the NADPH (100 micronM) indicates that they function as electron carries from the flavoprotein to molecular oxygen. As a corollary, Mitomycin-C and Carbazilquinone stimulated oxygen uptake by Ehrlich ascites tumor cells in the presence of glucose that Daunomycin and Adriamycin failed to do so, although the reason for it remains to be elucidated. Carbazilquinone, in contrast to others, also stimulated the microsomal NADH oxidation.


Late persistence of serum gamma-glutamyl transpeptidase activity after mononucleosis. Report of 3 cases. gamma-Glutamyl transpeptidase (GGTP) is a sensitive but nonspecific index hepatobiliary disease. In infectious mononucleosis (IM) or the mononucleosis-like disease attributable to cytomegalovirus (cytomegalovirus-induced IM), GGTP reverted to normal later than aspartate aminotransferase and alkaline phosphatase. In three cases elevated serum GGTP activity persisted for up to 24 months -- raising the question of persistent 'post-IM' hepatitis. Such prolonged GGTP activity was unusual in other late IM specimens. Possible, but unlikely, causes for such persistent GGTP activity are an unusual degree of hepatic damage during acute IM, excessive induction of microsomal enzyme system activity by drugs, or unusual Epstein-Barr virus carrier state activation that might contribute to ongoing hepatic structural damage. Other markers of chronic hepatocellular disease including aspartate aminotrasferase, alkaline phosphatase, and bilirubin were normal in late specimens from these 3 patients. The cause of their persistent elevated GGTP activities remains unknown.


Medical management of Crohn's disease in adolescence. The therapy of Crohn's disease in adolescence must balance the natural disease history of growth suppression, debilitation, and progression against possible drug-related adverse effects on growth and development. In contrast to published guidelines which usually suggest episodic and symptomatic treatment of relapses, we have attempted to suppress disease activity throughout adolescence. Sixteen consecutive adolescent patients treated with continuous medical therapy for a mean duration of 3.5 years are presented. Fourteen received long term prednisone therapy for maintenance of disease suppression. All 16 have been asymptomatic or have had only mild symptoms which did not interfere with regular activities. Only 1 subject had to be rehospitalized. He subsequently underwent bowel surgery. Aternate day corticosteroid administration has been attained in 11 patients; 10 are growing and developing at a normal rate. In total, 13 of 16 have achieved pubertal development appropriate for age. The 8 patients with distal ileal disease have had a consistently excellent response to medical therapy. There have been no major adverse effects from drug therapy. It is concluded that an effort to suppress disease activity continuously in adolsecents with Crohn's disease is warranted. Excellent symptomatic control and normal rate of growth can be expected in patients with primarily ileal disease.


[Effect of the cell dose on the development of 2 transplantable tumors in a syngeneic host]. The effects of different cell dose inoculation of plasmocitoma MOPC-315 and ADK-1t in Balb/c syngeneic mice are discussed in this work. Inoculated cell dose and neoplasia percent incidence have been noticed to be closely related, but unexpectedly two doses exist for each tumour, a comparatively small one and a definitely larger one, which cause nearly the same percent incidence. This paradox phenomenon is discussed also with reference to other researchers remarkers.


[Programmed breech deliveries (author's transl)]. The question whether the termination of a breech pregnancy by a programmed breech delivery would reduce the fetal risk was investigated. In 71 of 433 singleton breech deliveries (16%) the breech delivery was induced by oxytocin infusion. There were 38 primigravidas and 33 multi-gravidas. The Apgar and pH values showed the same results as in 3904 vertex deliveries with spontaneous onset of labour. The duration of labour was shortened. The incidence of Caesarean Section in programmed breech deliveries was 9.86%. All 71 infants were mature and healthy. There were no perinatal deaths. The results show that the fetal risk in breech deliveries is reduced by programmed breech delivery to the same risk as in vertex deliveries with spontaneous onset of labour.