Effect of S-adenosylhomocysteine on sulfhydryl xenobiotic transmethylases in rat liver

Rat liver cytosolic thiopurine methyltransferase and microsomal thiol methyltransferase were each found to be subject to control by the absolute molar ratio of S-adenosylmethionine to S-adenosylhomocysteine using cell-free enzyme preparations. As this ratio was lowered, inhibition of both sulfhydryl xenobiotic transmethylases occurred. On the other hand, when the ratio was decreased in vivo by the administration of D,L-homocysteine thiolactone to animals, this alteration was accompanied by an inhibition of only thiopurine methyltransferase activity. Thiol methyltransferase activity was not significantly affected after drug treatment, which would suggest that there is a compartmentalization of S-adenosylhomocysteine in the intact hepatocyte.     

Defective production of anti-herpes simplex virus antibody by neonatal mice. Reconstitution with Ia+ macrophages and T helper lymphocytes from nonimmune adult syngeneic mice

Both neonatal humans and mice are exquisitely susceptible to severe HSV infection. We have now documented a profound defect in the ability of neonatal C57BL/6 mice to produce anti-HSV ADCC antibody. This ability is acquired over the first 2 to 4 wk of life. Reconstitution of neonatal mice by i.p. injection of peritoneal cells from adult nonimmune syngeneic mice both affords dose-dependent protection against lethal HSV infection and reconstitutes the antibody-production defect. By cell-separation techniques (adherence, nylon wool column purification, B cell panning) and cell ablation techniques (silica treatment, irradiation, anti-T cell, anti-Ia, anti-Lyt-1.2 and anti-Lyt-2.2 monoclonal antibodies plus complement treatment) the subpopulations involved in the antibody production reconstitution of neonatal mice by adult cells were identified. These include both an Ia+, radioresistant, adherent, silica-sensitive macrophage population and a nylon wool column-purified, radiosensitive, anti-T, anti-Lyt-1.2-sensitive helper T cell population. The latter cell may be substituted for by concanavalin A-stimulated lymphokine-containing spleen cell supernatants or human recombinant IL 2. In addition to reconstitution of ADCC antibody production, the same cell populations, or cells plus lymphokine-containing supernatants or IL 2, protected the newborn mice from lethal HSV infection. Further characterization of this system and of soluble replacement factors has implications for therapy or immunoprophylaxis of human neonates with, or at risk of, HSV infection.     

Antibody-dependent cellular cytotoxicity and natural killer cytotoxicity of peritoneal cells from nude mice to herpes simplex virus-infected cells

Nude BALB/c mice (athymic) were more susceptible to fatal herpes simplex virus (HSV) than normal BALB/c mice (P = 0.002). The peritoneal cells of nude mice mediated levels of antibody-dependent cellular cytotoxicity (ADCC) of equal or greater magnitude than cells from normal BALB/c, heterozygote nu/+, or C57BL/6 mice. Unstimulated natural killer cytotoxicity of peritoneal cells from nude mice was higher (P less than 0.05) than that mediated by cells from C57BL/6 mice. Nude mice failed to make anti-HSV ADCC antibody 6 to 14 days post HSV inoculation, at times when nu/+, BALB/c, and C57BL/6 mice produced antibody. Passive reconstitution of nude mice with high titer intraperitoneal anti-HSV immune globulin provided circulating anti-HSV ADCC antibody and significant protection against lethal HSV infection.     

Identification of metabolites of phenylacetic acid in rat brain by plasma desorption mass spectrometry

Mass spectra of acyl hydroxamates may be obtained directly, without prior derivatization or gas chromatography, by the new technique of plasma desorption mass spectrometry. Authentic alkyl and aryl hydroxamates showed the expected protonated molecular ions when isobutane was used as the carrier gas. Advantage was taken of this novel technique to identify phenylacetyl-CoA and phenylbutyric acid in brain extracts obtained from young rats after a loading dose of phenylacetic acid. The formation of these metabolic products lends strong support to our suggestion that brain dysfunction induced by phenylacetic acid in experimental phenylketonuria may be due to decreased availability of CoA and acetyl-CoA in the rapidly developing brain.     

Effect of feed deprivation on hepatic membrane and lipoprotein fluidity and binding of lipoproteins to hepatic membranes in the chick (Gallus domesticus)

1. Male chicks were deprived of feed for 48 hr to study the effect of metabolic stress on hepatic membrane and lipoprotein fluidity and binding of radioiodinated lipoproteins to hepatic membranes. 2. Plasma levels of low density lipoprotein (LDL) and high density lipoprotein (HDL) were markedly and slightly elevated, respectively. 3. There was a reduction in lipoprotein and hepatic membrane fluidity. 4. Binding of [125I]LDL, but not [125I]HDL, to hepatic membranes was decreased. 5. It is suggested that a reduction in the fluidity of LDL and/or hepatic membranes impedes LDL catabolism in vivo.     

Merck Frosst Award Lecture 1998. Molecular dissection of the human multidrug resistance P-glycoprotein.

The human multidrug resistance P-glycoprotein is an ATP-dependent drug pump that extrudes a broad range of cytotoxic agents from the cell. Its physiological role may be to protect the body from endogenous and exogenous cytotoxic agents. The protein has clinical importance because it contributes to the phenomenon of multidrug resistance during chemotherapy. In this review, we discuss some of the results obtained by using molecular biology and protein chemistry techniques for studying this important and intriguing protein.     

Streptokinase therapy of myocardial infarct dependent on the severity of the disease]

A critical analysis of 14 clinical studies published since 1963 on the fibrinolytic treatment of acute cardiac infarct with streptokinase or urokinase shows that only half of them corresponds to the demands of a controlled clinical trial of therapy. The degree of seriousness of the infarct groups described in the different papers shows considerable fluctuations. The findings on letality are not uniform, there is a distinctive trend towards a reduced letality under streptokinase. It was only in recent studies that the dependence of the results of treatment on the seriousness of the disease were checked by retrospective stratification (Australian study) or by prospective stratification (European co-operative study). According to the findings of the European co-operative study those patients who are selected for the purpose of examination and affected with an infarct of medium degree of seriousness being not older than 12 hours have a significantly reduced six months letality under streptokinase on the conditions of this study.     

Functional relationship between receptor binding and biological activity for analogs of mammalian and salmon gonadotropin-releasing hormones in the pituitary of goldfish (Carassius auratus)

The relationship between gonadotropin-releasing hormone (GnRH) receptor binding and biological activity in the goldfish pituitary for mammalian and salmon GnRH (sGnRH) analogs with structural modification at the C terminus involving replacement of glycine amide with an alkyl amine and replacement of the Gly6 residue with D amino acids was examined. The GnRH receptor binding data were analyzed with a computerized curve-fitting program (LIGAND) for a single as well as two classes of binding sites; analysis based on one site fit estimated binding affinity and capacity for one class of binding site, and analysis based on two-site fit estimated binding affinity and capacity for two classes of binding sites (high-affinity/low-capacity and low-affinity/high-capacity binding sites). The estimated receptor affinity values were then used to determine the correlation between binding affinity and gonadotropin (GTH)-release potency in vitro. The highest correlation between biological activity and receptor binding affinity was obtained for the high-affinity/low-capacity binding sites and GnRH analogs containing Trp7 and Leu8 residues (i.e., the salmon GnRH structural format) (R = 0.940 +/- 0.150). For the same group of GnRH analogs, there was no significant correlation between the relative GTH-release potency and binding affinity of the low-affinity/high-capacity sites (R = 0.159 +/- 0.434), or that obtained from a one-site fit (R = 0.198 +/- 0.431). Similarly, for mammalian GnRH analogs, significant correlation between binding affinity and biological activity (R = 0.406 +/- 0.049) was only obtained for the high-affinity sites, although the degree of correlation was significantly lower than that obtained for salmon GnRH analogs. The present findings provide strong support for the hypothesis that high-affinity GnRH receptors are involved in the control of GTH release in the goldfish pituitary. In addition, the results demonstrate clearly that the presence of Trp7, Leu8 residues in salmon GnRH molecule, a native peptide in goldfish, is important for recognition of the ligand by the GnRH receptors in the goldfish pituitary, and that structural modifications at positions 6 and 10 in this peptide can increase receptor binding affinity and biological activity at the pituitary level. The most active sGnRH analog identified to date is [D-Arg6, Pro9-NEt]-sGnRH.