Solvent influence on model oxidations of alpha-tocopherol.

(+/-)-alpha-Tocopherol has been oxidised with t-butyl hydroperoxide in chloroform in order to simulate in vivo oxidations due to lipid hydroperoxides. t-Butyl hydroperoxide proved to be a weak oxidant and failed to oxidise alpha-tocopherol in 3 h at 60 degrees C. Inclusion of a small amount of ethanol in the reaction mixture brought about immediate oxidation and the formation of a new product, 5-ethoxymethyl-7,8-dimethyltocol in addition to the spiro dimer and spiro trimer of alpha-tocopherol, alpha-tocopherylquinone and 5-formyl-7,8-dimethyltocol. Formation of 5-ethoxymethyl-7,8-dimethyltocol increased with increasing concentrations of ethanol, up to a maximum of 59% at 20% ethanol. Further increase in ethanol concentration brought about a decrease in the oxidation of alpha-tocopherol and in the formation of 5-ethoxymethyl-7,8-dimethyltocol. Oxidation of the tocopherol model compound 2,2,5,7,8-pentamethyl-6-hydroxychroman under similar conditions produced the analogous product, 5-ethoxymethyl-2,2,7,8-tetramethyl-6-hydroxychroman together with 5-formyl-2,2,7,8-tetramethyl-6-hydroxychroman and 2-(3'-hydroxy-3'-methylbutyl)-3,5,6-trimethylbenzo-1,4-quinone.

Spectrophotometric assays for warfarin sodium and dicumarol. Use in rapid detection of patients suspected of drug intoxication or surreptitious noningestion.

The usefulness of assays for the rapid identification and determination of quantitative plasma levels of warfarin sodium and dicumarol is documented by the case histories of five patients: a man who accidentally took dicumarol for several weeks and developed an acute condition within the abdomen, a man who ingested 500 mg of warfarin sodium in a suicide attempt, a malingering nurse who surreptitiously took dicumarol, a nurse with warfarin intoxication who did not follow dosage prescription because of fear of developing thrombosis, and a woman with calf vein thrombosis who did not ingest the administered warfin sodium becausing of fear of developing bleeding. In every patient, the diagnosis of surreptitious ingestion or noningestion was suspected on the basis of the coagulation profile pattern. Confirmation of the disorder by the spectrophotometric assay enable the clinician to choose the appropriate course of action with speed and purpose.

The hemostatic mechanism after open-heart surgery. III. Correlation between the appearance of an abnormal protein demonstrated by gel electrophoresis and of an inhibitor of the extrinsic coagulation system (PEC).

A prospective study in 13 patients undergoing open-heart surgery with extracorporeal circulation revealed a marked decrease of the mean one-stage prothrombin time activity from 88% to 54% (p less than 0.005) but lesser decreases of factors I, II, V, VII and X. This apparent discrepancy was due to the appearance of an inhibitor of the extrinsic coagulation system, termed PEC (Protein after Extracorporeal Circulation). The mean plasma PEC level rose from 0.05 U/ml pre-surgery to 0.65 U/ml post-surgery (p less than 0.0005), and was accompanied by the appearance of additional proteins as evidenced by disc polyacrylamide gel electrophoresis of plasma fractions (p less than 0.0005). The observed increases of PEC, appearance of abnormal protein bands and concomitant increases of LDH and SGOT suggest that the release of an inhibitor of the coagulation system (similar or identical to PIVKA) may be due to hypoxic liver damage during extracorporeal circulation.

The oestrogenicity of equol in sheep.

The effects of intramuscular injection of synthetic racemic equol (+/- 3-(4-hydroxyphenyl)-3,4-dihydro-2H-1-benzopyran-7-ol) into wethers have been examined with respect to maintenance of plasma level, teat growth rate and the activity of the respiratory enzyme glucose-6-phosphate dehydrogenase. At a dose rate of 1.03 mmol/day a steady rise in 'total' (free plus conjugated) equol in plasma occurred to 1.78 mumol/l in 4 days. A dose rate of 2.07 mmol/day produced only a further slight increase in plasma equol. At a lower dose rate of 0.52 mmol/day the plasma concentration reached 0.62 mumol/l in 2 days and this was not exceeded thereafter. At the dose rate of 1.03 mmol/day over 7 days significant increases in teat length and glucose-6-phosphate dehydrogenase activity occurred but no significant changes were observed at the dose rate of 0.52 mmol/day. It appears that threshold levels of intake of equol which maintain a plasma level of about 1.65 mumol/l are needed for oestrogenic effects to become apparent within a relatively short time. Administration of 1.03 mmol/day over 5 days to ovariectomized ewes produced significant increases in uterine weight equivalent to those produced by 92 nmol stilboestrol dipropionate. Thus stilboestrol was apparently 56 000 times more potent than racemic equol.

The hemostatic mechanism after open-heart surgery. I. Studies on plasma coagulation factors and fibrinolysis in 512 patients after extracorporeal circulation.

Blood coagulation screening profiles were performed in 512 patients who underwent open-heart surgery with extracorporeal circulation. Severe coagulation disorders were found in 29 (5.6 per cent) patients. The most common abnormalities were low one-stage prothrombin time (PT) activities and impaired whole blood clot retractions. In the majority of patients the discrepancy between low PT's and normal or only slightly depressed factor II, V, VII, and X activities was explained by the presence of an inhibitor of the extrinsic system. Eight patients demonstrated the heparin rebound phenomenon but only 1 bled excessively. The pattern of severe hepatic dysfunction was found in 4 and severe depression of vitamin K-dependent factors due to oral anticoagulants in 2. Two had disseminated intravascular coagulation. Seventeen patients with normal coagulation screening profiles bled excessively postoperatively. Of these, 2 had moderate thrombocytopenia associated with a marked platelet functional abnormality. Revision of the wound in 13 revealed a surgical hemostatic defect and in 2 the cause of bleeding could not be determined.

Meanings of political participation among black and white women: political identity and social responsibility.

This study examined the correlates of midlife political participation among 64 Black and 107 White women of the college classes of 1967-1973. Compared with White women, Black women scored higher on political participation, generativity, power discontent, and politicization. Factor analysis of personality and political attitude variables yielded three factors labeled Political Identity, Power Discontent, and Social Responsibility. Adult political participation was regressed on level of student activism and index scores of political identity, power discontent, and social responsibility. For both racial groups, social responsibility was associated with midlife political participation. For White women, political identity was also related; for Black women, student activism bore a significant relationship. The findings suggest that Black and White women's historical and political contexts imbued their political activities with different meanings.

Structure and solution equilibria of D-glucose and D-mannose sulfite adducts.

An X-ray crystallographic study has confirmed that the potassium bisulfite adducts of D-glucose and D-mannose have open-chain structures with R and S configurations respectively at C-1. NMR studies have shown that each sugar gives rise to two bisulfite compounds, and solution-state structures and conformations of these isomers have been deduced from analysis of (1)H NMR spectra. (13)C NMR data for the four adducts are given. Furanose forms of the D-glucose and D-mannose have been detected in the equilibrium solutions.

Review of the analytical chemistry of melatonin.

A survey has been made of procedures, chemical and biochemical, for the separation and estimation of melatonin, the active constituent of the pineal gland. In view of the extremely small amounts of material generally present, discussion is built around chromatographic and spectroscopic techniques leading to a comparison with assays based on the skin-blanching effect of the hormone.

Chemical composition of essential oil from ripe fruit of Schinus terebinthifolius Raddi and evaluation of its activity against wild strains of hospital origin.

The essential oil (EO) composition of ripe fruit of S. terebinthifolius Raddi was analyzed by GC-MS. The oil extraction yielded 6.54 ± 1.06% (w/w). Seventeen compounds were identified, accounting for 91.15% of the total oil, where monoterpenes constituted the main chemical class (85.81%), followed by sesquiterpenes (5.34%). The major monoterpene identified was δ-3-carene (30.37%), followed by limonene (17.44%), α-phellandrene (12.60%) and α-pinene (12.59%). Trans-caryophyllene (1.77%) was the major sesquiterpene identified. The antibacterial activity of the essential oil was evaluated against wild strains of hospital origin (Escherichia coli, Pseudomonas sp., Klebsiella oxytoca, Corynebacterium sp., Staphylococcus aureus, Enterobacter sp., Enterobacter agglomerans, Bacillus sp., Nocardia sp. and Streptococcus group D). The essential oil of the ripe fruit of S. terebinthifolius Raddi has shown to be active against all tested wild strains, with minimum inhibitory concentration ranging from 3.55 µg/mL to 56.86 µg/mL. However, it has revealed some differences in susceptibility: the general, Gram-positive species showed greater sensitivity to the action of EO, which is probably due to the lower structural complexity of their cell walls.

Chemical composition of essential oil from ripe fruit of Schinus terebinthifolius Raddi and evaluation of its activity against wild strains of hospital origin

The essential oil (EO) composition of ripe fruit of S. terebinthifolius Raddi was analyzed by GC-MS. The oil extraction yielded 6.54 ± 1.06% (w/w). Seventeen compounds were identified, accounting for 91.15% of the total oil, where monoterpenes constituted the main chemical class (85.81%), followed by sesquiterpenes (5.34%). The major monoterpene identified was δ-3-carene (30.37%), followed by limonene (17.44%), α-phellandrene (12.60%) and α-pinene (12.59%). Trans-caryophyllene (1.77%) was the major sesquiterpene identified. The antibacterial activity of the essential oil was evaluated against wild strains of hospital origin (Escherichia coli, Pseudomonas sp., Klebsiella oxytoca, Corynebacterium sp., Staphylococcus aureus, Enterobacter sp., Enterobacter agglomerans, Bacillus sp., Nocardia sp. and Streptococcus group D). The essential oil of the ripe fruit of S. terebinthifolius Raddi has shown to be active against all tested wild strains, with minimum inhibitory concentration ranging from 3.55 μg/mL to 56.86 μg/mL. However, it has revealed some differences in susceptibility: the general, Gram-positive species showed greater sensitivity to the action of EO, which is probably due to the lower structural complexity of their cell walls.

Alteration of the kinetics of thrombin-catalyzed hydrolysis of amino acid ester substrates by sodium cholate and other steroids.

Thrombin-catalyzed hydrolysis of TAME proceeds by an initial zero-order phase which later falls off into an apparent first order reaction as substrate becomes limiting. Optimum amounts of sodium cholate not only accelerated TAME hydrolysis but also altered its kinetics to apparent zero-order to complete substrate hydrolysis. As this implies, the rate of hydrolysis in the presence of cholate was found to be independent of substrate concentration, provided concentrations TAME and cholate were low enough to prevent precipitation of some TAME-cholate as an insoluble complex. The formation of a soluble complex composed of polymeric molecules of TAME and cholate may explain both the acceleration and the change in reaction order. Although the pH and temperature optima for TAME hydrolysis by thrombin rising pH and temperature on the ascending protion of the curves. This is believed to be due to the greater solubility of the TAME-cholate complex. The effects of cholate on thrombin-catalyzed hydrolysis of other arginine esters as well as esters of lysine, histidine and phenylalanine were also studied. Solutions of sodium desoxycholate and androsterone-3-sulfate accelerated TAME hydrolysis as did supensions of testosterone, etiocholanolone, androsterone, androsterone-3-hemisuccinate and pregnandiol-3-glucuronidate. However, isoandrosterone, progesterone, pregnandiol, estradiol, estrone, estriol, estrone-3-sulfate, cholesterol, corticosterone, hydrocortisone and hydrocortisone-3-phosphate had no significant effect on TAME hydrolysis by thrombin. The ability of the andorogenic hormones to accelerate hydrolysis appeared to depend to some extent on the configuration of the substituent group at C3 and the hydrogen at C5. Androsterone-3-hemisuccinate was, like cholate, able to accelerate the hydrolysis of TAME at apparent zero-order kinetics to complete substrate hydrolysis.

Purification and properties of a plasminogen activator from pig heart.

An improved procedure is described for the purification of plasminogen activator from pig heart. The initial purification steps were similar to these described previously (Bachmann, F., Fletcher, A. P., Alkjaersig, N., and Sherry, S. (1964) Biochemistry 3, 1578--1585). Use of a novel extraction medium containing EDTA, cysteine, and 2,3-dimercaptopropanol-1 facilitated the removal of large amounts of inert proteins prior to gel filtration on Bio-Gel P-150. The final product had a specific activity of 120,000 to 160,000 CTA units/mg of protein (CTA, Committee on Thrombolytic Agents of the National Heart Institute). Total purification over pig heart was 25,000 to 30,000-fold, average recovery compared to the initial extract was 6 to 8%. Polyacrylamide gel electrophoresis revealed a major and two minor components. The molecular weight of the activator determined by gel filtration was 51,500 +/- 3,400 for the major activity component and 48,000 for a minor component which was partially separated from the major peak in eight of nine chromatography runs. A gamma-globulin fraction of antiserum against purified activator neutralized the biological activity of the activator on fibrin plates. Immunoelectrophoresis of gel-filtered activator revealed only one anodic component.

The effect of spicules obtained from sickle red cells on clotting activity.

Spicules from sickle red cells were examined for their effects on the clotting activity of blood. The spicules were obtained from the sickle red cells after deoxygenation and oxygenation and were tested for clotting activity with Russell's viper venom assay. A marked increase in clotting activity was observed when spicules were added to the system. The increase was distinctly greater than that observed after the addition of sickle red cells while normal red cells had little effect. Vesicles prepared from sickle or normal red cells by incubation with the ionophore A-23187+Ca2+ also markedly increased clotting activity. The effect of spicules or vesicles on the clotting system may be related to reorganization of phospholipid in the spectrin-poor membrane of the spicules or vesicles. Because of these effects, the spicules from the sickle red cells may contribute to the hypercoagulable state in these patients and possibly to their vaso-occlusive crises since free spicules are present in their plasma. Vesicles from red cells from other types of anaemia with hypercoagulability may have a similar effect on coagulation.

Long-term frequent plasma exchange donation of cryoprecipitate.

Plasma exchange donation accomplishes the selective donation of cryoprecipitate. It facilitates the repeated donation of large quantities of factor VIII by individual donors and reduces donor exposure for recipients. A highly motivated donor is described who has undergone 103 donations between May 1983 and March 1987, producing 359,460 IU of factor VIII and supplying all the factor VIII needed since August 1983 by his severely affected hemophiliac son, now age 14. The donor has remained in good health, and no significant abnormalities have been noted in hematologic, biochemical, immunologic, coagulation, and serum protein testing. Extensive experience with this donor suggests that repeated plasma-exchange donation is safe and can sometimes allow single-donor support of severe hemophiliacs.