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D K Liu

Publications and source records attributed to D K Liu.

At least 55 records · Page 3Linked to original sources

Isolation of ribonuclease-free intact polyribosomes from rat kidney.

High levels of RNAase present in rat kidney have prevented isolation of intact polyribosomes from this tissue. This problem has been circumvented by a thorough in situ arterial perfusion of rat kidney, coupled with homogenization of the perfused rat kidney in heparin and detergents-fortified high-speed supernatant prepared from rat liver. This procedure reduced RNAase activity in the homogenate by as much as 70%. Sedimentation of the polyribosomes from this homogenate through a layer of 2.0 M sucrose resulted in a 78--80% yield of polyribosomes from the rat kidney. The resulting polyribosomal pellet contained less than 8% of the RNAase activity present in polyribosomes from non-perfused rat kidney. The remaining RNAase activity was separated from the larger polyribomes by sucrose density gradient centrifugation. The majority of the polyribosomes were larger than tetramers. This procedure also incrased both the yield and size of polyribosomes from rat and mouse liver.

Animals↗

Ca2+-stimulated ribonuclease. A new marker enzyme of differentiated rat mammary tissues.

A unique ribonuclease, active only in the presence of Ca2+, was present in lactating mammary gland and milk of the rat. This enzyme was absent from virgin-rat mammary gland and non-mammary tissues of lactating rats. The presence of moderate activity in differentiated mammary tumours, together with an increase in activity in normal tissue parelleling development of mammary function, identify this enzyme as a marker of mammary differentiation in the rat.

Animals↗

Regional differences in ribonuclease content of rat and mouse kidney.

Kidney cortex, red medulla and white medulla were separated into nuclei, mitochondria, microsomal and 105000g supernatant fractions. Assay of RNAase (ribonuclease) activity at pH7.8 revealed that, for each subcellular fraction, activity was much greater in cortex than in red or white medulla; this was true for both free RNAase and total (free plus latent) RNAase. For example, the free RNAase activity in the 105000g supernatant of cortex was 5 and 8 times higher than in red and white medulla respectively. No latent RNAase activity was found in any particulate fraction. Latent supernatant RNAase activities (suggesting presence of bound RNAase inhibitor) were similar in cortex and medulla. The cortex supernatant contained minimal free RNAase inhibitor, whereas that of the red and white medulla showed about one-third and one-tenth respectively of the inhibitor activity measured in liver. Adrenalectomy did not change RNAase activity in any fraction nor the content of free RNAase inhibitor in the kidney supernatant, but did decrease the liver RNAase inhibitor content by 40%. In supernatants from mouse kidney, both free and total RNAase activities of both cortex and red medulla were similar to those of rat red medulla. Mouse cortex contained appreciably higher amounts of free RNAase inhibitor than rat cortex. The difference between the rat and mouse cortical RNAase activity and inhibitor content may help explain the relative ease with which satisfactory renal polyribosome profiles were obtained from mouse kidneys. Our results, as well as those of Kline & Liberti [(1973) Biochem. Biophys. Res. Commun.52, 1271-1277], showing that renal red and white medulla are more active than cortex in protein synthesis, are consistent with the hypothesis that the RNAase-RNAase-inhibitor system may participate in the regulation of protein synthesis.

Adrenalectomy↗

Transfer RNA and aminoacyl transfer RNA in developing rats.

The total transfer RNA (tRNA) level in the liver, kidney, skeletal muscle and heart muscle of developing rats was determined by purification using (3H)tRNA as an internal standard. Liver and kidney contained almost twice as much tRNA per gram tissue as heart and skeletal muscle. There were no apparent differences between the sexes. The aminoacylation capacities of six tRNA species (alanyl, aspartyl, leucyl, methionyl, phenylalanyl, and tryptophanyl) from rat liver were not different during 3 developmental stages (suckling, weaning and young adult), and there were also no differences noted between males females. The in vivo percent aminoacylation of 4 tRNAs (aspartyl, leucyl, methionyl, and phenylalanyl) was lower during the newborn and suckling periods than in weaning and young adult rat livers. The tRNA of young adults was almost completely aminoacylated in vivo with the exception of alanyl-tRNA.

Acylation↗

Environmental and genetic factors affecting the response of laboratory animals to drugs.

Only some of the diverse factors that can affect drug disposition and response in laboratory animals have been identified at the present time. These numerous factors contribute to large day-to-day variations that have become a major problem impeding investigation of drug disposition and response in laboratory animals. Although these variations render many experiments difficult to interpret and produce large discrepancies in the literature, few published investigations using laboratory animals provide sufficient details to permit replication of the studies under similar conditions with respect to these variables. Thus, the importance of these variables in affecting results is apparently insufficiently recognized at present. Two commonly overlooked variables affecting the activity of hepatic microsomal enzymes (HME) in rodents and hence the rate at which rodents eliminate from their bodies many foreign compounds are the bedding under the wire mesh cage and the relative cleanliness of the environment. Numerous chemicals present in relatively low concentrations in the environment of the animal room can significantly alter HME activity. Representative of these chemicals are aromatic hydrocarbons in cedarwood bedding, eucalyptol from aerosol sprays, and chlorinated hydrocarbon insecticides, each of which induces HME activity, whereas ammonia generated from feces and urine accumulated in unchanged pans under cages may inhibit HME activity. Chloroform, identified as an environmental contaminant of the water and air of certain cities, exhibits sex and strain differences with respect to toxicity (LD50) in mice. After intraperitoneal injection, twice as much chloroform accumulated in the kidneys of males from the sensitive strain (DBA/2J) as from the resistant (C57BL/6J) strain. First generation offspring were midway between parental strains both with respect to LD50 and renal accumulation of chloroform.

Animals↗

Genetic control of chloroform toxicity in mice.

Mouse strain differences suggest intermediate or multifactorial gentic control of chloroform-induced renal toxicity and death. The chloroform dose lethal to 50 percent of animals was four times higher in C57BL/6J males than in DBA/2J males. Twice as much chloroform accumulated in the kidneys of the sensitive as the resistant strain. First generation offspring were midway between parental strains for both parameters.

Androgens↗

Alkaline ribonuclease and ribonuclease inhibitor in mammary gland during the lactation cycle and in the R3230AC mammary tumour.

Alkaline RNAase (ribonuclease) and RNAase inhibitor were assayed to determine the potential role of the degradative process in regulating the amount of RNA in the mammary gland and mammary tumour. Very little free alkaline RNAase activity was found in the cytosol fraction of the mammary gland of virgin, pregnant, lactating or involuting Fischer rats. However, addition of p-chloromercuribenzoate to the assay medium revealed latent RNAase which, when expressed on a DNA basis, decreased during pregnancy and lactation. The cytosol latent RNAase is stable in 0.125 M-H2SO4. The non-cytosol RNAase activity also decreased during pregnancy and lactation. Addition of Triton X-100 produced slightly higher activity at all stages tested. The inhibitor activity in rat mammary gland was very low before pregnancy, increased gradually during pregnancy and more dramatically at parturition, continued to increase throughout lactation and returned to resting-gland values by the sixth day of involution. The increase during pregnancy may be due to the increased cellularity of the gland, whereas the gain during lactation was more than could be accounted for by increases in cell number. The R3230AC transplantable mammary tumour resembles the normal gland in early lactation with respect to both its cytosol and non-cytosol alkaline RNAase activities and its moderately high content of RNAase inhibitor. The relatively high inhibitor and low RNAase activities in both the gland of the lactating rat and in the tumour are of potential significance in maintaining high amounts of RNA and increased rates of protein synthesis in these tissues.

Animals↗