Isotopic analysis of the reaction catalyzed by glycerol dehydrogenase

Glycerol dehydrogenase catalyzes the reversible NAD(+)-dependent oxidation of glycerol to form dihydroxyacetone. Initial velocity, product, and dead-end inhibition studies performed for the forward and reverse reactions support an ordered kinetic mechanism with NAD+ binding first and NADH released last. A monovalent cation is required for enzymatic activity and glycerol binding, with K+ having the highest activity as measured by V. The pH dependence of the kinetic parameters V and V/Kglycerol, as well as the temperature dependence of the V pH profile, suggested that an enzymic carboxylate group functions as a base in catalysis. The pH dependence of the primary deuterium kinetic isotope effect shows that DV/Kglycerol increases from a pH-independent value of 1.15 at high pH values to a pH-independent value of 2.44 at low pH values. DV exhibits a similar pH dependence, increasing from a pH-independent value of 2.57 at high pH values to a pH independent value of 4.88 at low pH values. A chemical mechanism for enzymatic glycerol oxidation is proposed based on the data.     

von Willebrand factor as a regulator of intrinsic factor X activation

Factor VIII is an important cofactor in the intrinsic activation of factor X. To function effectively as a cofactor, factor VIII must be activated. In plasma, factor VIII circulates in a complex with von Willebrand factor, and although thrombin can activate complexed factor VIII, the activation by activated factor X is inhibited by von Willebrand factor. In this study, the effect of von Willebrand factor on the generation of factor Xa by the factor IXa-VIII complex was investigated. Purified human factors VIII, IXa, and X were incubated on human umbilical vein endothelial cells or phospholipid vesicles in the presence of calcium ions, and the generation of factor Xa was followed. In the presence of von Willebrand factor, a prolonged lag-phase and a dose-dependent inhibition of factor X activation was observed. These effects were not observed when von Willebrand factor was preincubated with a monoclonal antibody directed against von Willebrand factor that blocks factor VIII binding. When factor VIII was activated with thrombin before the incubation, neither the monoclonal antibody nor von Willebrand factor had an effect on the rate of factor X activation. Preincubation of endothelial cells with the monoclonal antibody resulted in a somewhat higher rate of factor X activation. When endothelial cells from a patient with von Willebrand's disease type I were used, preincubation of the monoclonal antibody had no effect on the rate of factor X activation. We conclude that von Willebrand factor on the surface of endothelial cells can modulate the intrinsic factor X activation. This effect is greatly enhanced, however, by the addition of exogenous von Willebrand factor.(ABSTRACT TRUNCATED AT 250 WORDS)     

Investigation of the sequence of colonization and candidemia in nonneutropenic patients

Among neutropenic patients with hematologic malignancies, candidemia has been shown to arise typically from autoinfection after colonization. In patients without neutropenia, we examined the similarities of strains colonizing or infecting various body sites and those subsequently causing Candida bloodstream infections. Strain similarity was examined by karyotyping and restriction endonuclease analysis of genomic DNA (REAG) by using two restriction enzymes (SfiI and BssHII). The banding patterns of 42 isolates from 19 patients were independently evaluated in a blinded fashion by three observers. The interobserver reliability measured with a generalized kappa statistic was 0.59 for karyotyping, 0.84 for REAG with SfiI, and 0.88 for REAG with BssHII (P < 0.001 for each). REAG classified the initial colonizing or infecting isolate and subsequent blood isolates as identical in 16 patients (84%). The mean duration of colonization or infection prior to a positive blood culture was 5 and 23 days in patients infected with related and unrelated isolates, respectively (P = 0.14; 95% confidence interval = -14.5 to 50.5). Karyotyping results matched the REAG results for isolates from 14 of the 19 patients (74%). In patients infected with identical isolates, the initial isolate was most frequently recovered from the urine (n = 5) or vascular catheter tips (n = 4). In the five subjects with organisms showing disparate results between the methods, karyotyping revealed different banding patterns, whereas REAG suggested that the isolates were identical. Candida colonization or infection with an identical strain frequently precedes bloodstream infection in nonneutropenic patients. Future studies should evaluate whether patients at high risk for candidemia and who have vascular catheter or urine samples that are positive for a Candida on culture should be treated empirically.     

Surgical inferior repositioning of the maxilla: treatment considerations and comprehensive management

Authors studied two different groups of patients. One of them with febrile seizures and the other febrile seizures associated to epilepsy, in order to find risk factors for the presentation of this one. They conclude that epilepsy has a presentation rate of 9,6% in patients with previous febrile seizures, having the highest risk when they show previous familial seizures, neonatal pathology and altered EEG after the first febrile convulsion. They study the role of phenobarbital taken after febrile seizures, as prophylaxis. No effect was observed in the prevention of following epilepsy.     

Inversion polymorphism in natural populations of Drosophila bipectinata

Inversion polymorphism was studied in seven natural populations of Drosophila bipectinata, five from northern and two from southern India. Chromosomal analysis of these populations revealed the presence of three paracentric inversions which are widespread in populations of D. bipectinata. Quantitative data indicated that the frequency of inversions and the level of inversion heterozygosity were very low in populations of D. bipectinata. There is no evidence for genetic differentiation between populations as a result of inversion polymorphism. These findings provide evidence for rigid chromosomal polymorphism in D. bipectinata. Significant non-random associations between inversions indicated epistatic interaction between inversions in natural populations of D. bipectinata.     

Noradrenaline-H3 uptake by isolated rat organs with active and blockaded adrenoreceptors

The beta-adrenoreceptor blocking agent propranolol was shown to activate noradrenaline-N3 absorption by the rat isolated organs by 30-180%. The alpha-adrenoreceptor blocking agent phentolamine only activated by 30-55% the noradrenaline-N3 absorption in the organs possessing the postsynaptic alpha-adrenoreceptors (ductus semen, spleen and amsll intestine). The beta-adrenoreceptor activator isopropylnoradrenaline decreased the intensity of noradrenaline-N3 absorption in all the organs by 15-50%. The alpha-adrenoreceptor activating agents (urine and mesaton) only depressed the noradrenaline-N3 absorption by 20-45% in the organs possessing postsynaptic alpha-adrenoreceptors. The data obtained suggest existence of reverse transsynaptic regulatory effects via adrenoreceptors of the cell-effector on the processes of noradrenaline absorption by adrenergic neuron.     

Chemiluminescence of plant tissues preserved in aldehydes and exposed to the salts of heavy metals

It was established that plant tissues killed by various aldehydes in low concentrations, which evoked general enzyme inactivation for a long time retain permeability and lipoprotein bonds in the cell membranes. Such preparations from Elodea Canadensis leaves are used for: Cu, Zn, Pb, Hg toxical effect investigation. It was stated by chemoluminescence methods that these ions induce slow developed oxidative radical chain reaction in lipid cell membranes and break the lipoprotein bonds.