Copper(II) (3,5-diisopropylsalicylate)2 oxidizes thiols to symmetrical disulfides and oxidatively converts mixtures of 5-thio-2-nitrobenzoic acid and nonsymmetrical 5-thio-2-nitrobenzoic acid disulfides to symmetrical disulfides

L-cysteine, D-penicillamine, and L-glutathione were oxidized to symmetrical disulfides in the presence of Cu(II)(3,5-DIPS)2 and air-oxygen at physiologic pH, 7.3. Air-oxygen caused the oxidation of thiol reduced copper, Cu(I), to Cu(II), as evidenced by expected spectrophotometric changes in these reaction mixtures. L-cysteine, D-penicillamine, and L-glutathione formed mixed disulfides and TNB with the addition of DTNB to solutions of these thiols. The observed order of reactivity for these thiols with DTNB was: L-cysteine greater than D-penicillamine greater than L-glutathione. Surprisingly, Cu(II)(3,5-DIPS)2 converted these mixed disulfides to their symmetrical disulfides and DTNB, and although the initial conversion rate was rapid, complete conversion required more than two hours. These observations suggest caution with regard to the spectrophotometric determination of thiols immediately after the addition of Ellman's reagent. These results also clarify an earlier report concerning the oxidation of thiols by Cu(II)(o-phenanthroline)2 and offer caution with regard to the determination of thiols using DTNB in the presence of copper complexes. Spectrophotometric data are provided in support of the suggestion that analysis of plasma or cellular samples for thiols be done in the absence of copper(II) complexes to avoid false negative results.     

Differential and analytical subfractionation of rat liver components internalizing insulin and prolactin

Receptor-mediated endocytosis of 125I-insulin and 125I-prolactin into liver parenchymal cells has been studied by quantitative subcellular fractionation. Differential centrifugation yielded three particulate fractions, N (nuclear), ML (large granule), and P (microsomes), and a final supernatant (S). Quantitative differences in the extent and rates of accumulation of 125I-insulin and 125I-prolactin into the fractions were observed. The acidotropic agent chloroquine and the microtubule disrupting agent colchicine were administered separately to rats. The agents increased significantly the T 1/2 of hormone clearance from the liver and augmented the accumulation of both ligands in the low-speed ML fraction. However, differences in the rates of accumulation of insulin and prolactin into all cell fractions were still maintained. Analytical centrifugation of each of the particulate fractions was carried out in order to determine if different endocytic components were specific to insulin or prolactin internalization. This was not the case. An "early" endosomal component of density 1.11 was identified in microsomes. A "late" endosome of density 1.10 was identified in the large granule (ML) fraction. Both endosomal components appeared to accumulate insulin and prolactin but at different rates. Marker enzyme analysis identified the presumed plasma membrane component in microsomes (density approximately 1.155). This component showed a significant difference in the rate of loss of 125I-insulin (T 1/2 approximately 4.1 min) as compared to that of 125I-prolactin (T 1/2 approximately 12.7 min). A further difference in the handling of the ligands was observed in early endosomes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cloning and expression of a cDNA encoding a catalytically active fragment of calf thymus DNA polymerase alpha

A calf thymus cDNA expression library was constructed in the EcoRI site of lambda gt11 and probed with an antibody raised against calf thymus DNA polymerase alpha. Three classes of antibody-reactive clones were isolated. The largest class carried a 1.9 kilobase calf cDNA insert and expressed a 165-175 kilodalton beta-galactosidase:calf fusion protein which displayed DNA polymerase activity. The characteristic responses of the polymerase activity to alpha-specific inhibitors and antibodies identified the 1.9 kilobase cDNA as a sequence specifically derived from the structural gene encoding the pol alpha catalytic core.     

Nitrification and organic nitrogen formation in soils

Summary The formation of mineral nitrogen species and of organic nitrogen was studied in three different types of soils in relation to the application of the nitrification inhibitor nitrapyrin. The results indicate that nitrification brings about a deficit in total mineral nitrogen and a concomitant surplus in non biomass organic nitrogen. This phenomenon increases with increasing levels of applied ammonium nitrogen and soil organic matter. The phenomenon is considered to be due to the reaction of the transient nitrite formed with soil phenolic compounds and appears to be of significance in all soils in which nitrification occurs, even neutral to alkaline and low carbon soils.     

Limnology and plankton periodicity of Jos Plateau water reservoir,Nigeria, West Africa

Limnological data (Dec. 1980–Jan. 1982) on the plankton and water chemistry of Lamingo Dam, located within the Jos biotite granite area of the Plateau State (Nigeria) are presented. The water-body falls in Beadle's (1981) category I of African lakes (conductivity < 40 µS cm^–1). Alkalinity (% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0Jf9crFfpeea0xh9v8qiW7rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGabmiEayaara% aaaa!3914!\[\bar x\] = 0.3 meq l^–1), principally composed of bicarbonates, dominated the anions % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0Jf9crFfpeea0xh9v8qiW7rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGaaK4waiaajI% eacaqIdbGaaK4tamaaxababaGaeyOeI0caleaacaaIZaaabeaakiaa% b6dacaqIdbGaaKiBaiaaj2cacaqG+aGaaK4uaiaaj+eadaqhaaWcba% GaaKinaaqaaiaajkdacaqITaaaaOGaaKyxaaaa!4657!\[\user1{[HCO}\mathop -\limits_3 {\text{ > }}\user1{Cl - }{\text{ > }}\user1{SO}_\user1{4}^{\user1{2 - }} \user1{]}\]The plankton were characterized by a moderate standing crop of phytoplankton, and zooplankton were, generally, very limited in species and abundance. The order of dominance for the categories of phyto and zooplankton was: [Bacillariophyceae > Chlorophyceae > Dinophyceae] and [Rotifera > Crustacea] respectively. A diel cycle was characterized by nocturnal upward migration of the zooplankton, and the reverse behaviour in the phytoplankton.Interrelations between the biotic assemblages of plankters and various physical and chemical variables are discussed.     

Influence of the presence of Zymomonas anaerobia on the conversion of cellobiose, glucose, and xylose to ethanol by Clostridium saccharolyticum

To convert sugar mixtures containing cellobiose, glucose, and xylose to ethanol in a single step, the possibility of using a coculture consisting of Clostridium saccharolyticum and Zymomonas anaerobia was studied. In monoculture, C. saccharolyticum utilized all three sugars; however, it preferentially utilized glucose and produced acetic acid in addition to ethanol. The formation of acetic acid from the metabolism of glucose inhibited the growth of C. saccharolyticum and, consequently, the utilization of cellobiose and xylose. In monoculture, Z. anaerobia utilized glucose at a rate of 50 g/L day, but it did not ferment cellobiose or xylose. In coculture, Z. anaerobia converted most of the glucose to ethanol during the lag phase of growth of C. saccharolyticum, which then converted cellobiose and xylose to ethanol. The use of this coculture increased both the rate and the efficiency of the conversion of these three sugars to ethanol, and produced relatively small amounts of acetic acid.     

Catechol-O-methyltransferase: A method for autoradiographic visualization of isozymes in Cellogel

An electrophoretic procedure for separating the molecular forms of catechol-O-methyltransferase in cellulose acetate gel is described; the zones of enzyme activity were revealed by autoradiography. The electrophoretic patterns of the enzyme in several tissues and cell lines derived from four different species are presented.This investigation was supported by Grant 500.6/Ric. 70/1981 from the Italian Ministry of Health and by the Italian Ministry of Education. We are grateful to Dr. M. Castagnola for useful advice and help with thin-layer chromatography.     

Decreased triiodothyronine binding to isolated nuclei from livers of preobese and obese (ob/ob) mice

L-Triiodothyronine (T3) binding to hepatic nuclei from (ob/ob) mice at different ages was examined and compared with that of lean controls. Results showed a significant reduction in T3 binding in liver nuclei of obese mice at all ages studied. The preobese mice at 2 weeks of age had 27.9% fewer receptor sites/mg DNA compared to lean controls, receptor concentration further decreased to 67.7% at 18 weeks of age. Data presented here demonstrates that the impaired triiodothyronine (T3) binding to hepatic nuclei present in older (ob/ob) obese mice is an antecedent to the obesity. This report also helps to explain the poor thermoregulation and low oxygen consumption present during the preobese phase of the postnatal development of these animals.     

Microvillar channels: a unique plasma membrane compartment for concentrating lipoproteins on the surface of rat adrenal cortical cells

Electron microscopic studies of perfused rat adrenals indicate that plasma lipoproteins become concentrated in a specialized cell surface compartment called microvillar channels. Closely associated plasma membranes of sinusoidal microvilli of zona fasciculata cells form channels that normally are filled with electron dense particles the size of high density lipoproteins (HDL). In rats made acutely deficient in plasma lipoproteins (by treatment with 4-aminopyrazolo[3,4-d]pyrimidine (4-APP) for 1 day), particles within the microvillar channels are decreased in number. When adrenal glands of these rats are perfused with media lacking plasma lipoproteins, many but not all of these HDL-like particles are washed out. However, when these adrenals are perfused with large amounts (100-500 micrograms protein/ml) of HDL, microvillar channels become packed with electron dense particles similar to those found in vivo. These microvillar channels become wider and filled with larger particles when low density lipoproteins (LDL) are perfused through the adrenals. Autoradiograms of 125I-labeled HDL-perfused adrenals show silver grains specifically associated with the cell surface microvillar channels, and confirm the notion that the particles filling the channels are exogenously delivered HDL. Physiologic data from similarly perfused adrenals in a parallel study show that the channel-refilling process is directly related to selective (i.e., nonendocytic) cholesterol uptake and that this cholesterol uptake is associated with corticosterone production. Together, these data suggest the hypothesis that plasma lipoprotein cholesterol utilized for corticosteroid synthesis in rat adrenal fasciculata cells may be derived from lipoproteins trapped in surface-associated microvillar channels. Although the mechanism responsible for the cholesterol transfer is not yet defined, it is clearly distinct from the classical process of receptor-mediated endocytosis and catabolism of lipoprotein particles.