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Effect of chenodiol on the small intestine. Unimpaired structure and function during therapy for gallstone dissolution.

To test whether long-term oral dosage with chenodiol (chenodeoxycholic acid) used for dissolution of cholesterol gallstones would cause impairment of small-intestinal structuree or function, ten patients were studied before and after three months of oral chenodiol administration, 15 mg/kg of body weight per day. Small-intestinal structure was assessed by roentgenogram and intestinal biopsy, using both light and electron microscopy. Small-intestinal function was assessed by xylose, fat and vitamin B12, lactose, and bile-acid absorption. Bile acid metabolism was also characterized by the breath test for deconjugation using carbon dioxide labeled with radioactive carbon 14. No significant abnormalities were found. The results suggest that oral chenodiol administration does not impair intestinal structur or function in doses used for gallstone dissolution.

Administration, Oral↗

Feasibility of a mitochondrial pyruvate malate shuttle in pancreatic islets. Further implication of cytosolic NADPH in insulin secretion.

Previous studies indicated that in pancreatic islets the amount of glucose-derived pyruvate that enters mitochondrial metabolism via carboxylation is approximately equal to that entering via decarboxylation and that both carboxylation and decarboxylation are correlated with capacitation of glucose metabolism and insulin release. The relatively high rate of carboxylation is consistent with the current study's finding that pyruvate carboxylase is as abundant in pancreatic islets as it is in liver and kidney. Since islets do not contain phosphoenolpyruvate carboxykinase and, therefore, cannot carry out glyconeogenesis from pyruvate, the carboxylase might be present in the islet to participate in novel anaplerotic reactions. This idea was first explored by incubating mitochondria from various tissues with pyruvate. Mitochondria from tissues, such as pancreatic islets, liver, and kidney, in which pyruvate carboxylase is abundant, exported a large amount of malate and little or no citrate, isocitrate, and aspartate to the medium. The amount of malate within the mitochondria was < 1% that in the medium. When pancreatic islet mitochondria were incubated with [1-14C]pyruvate, radioactive carbon appeared in the medium primarily in malate. Very little radioactivity appeared in amino acids, and little or no radioactivity appeared in citrate and isocitrate. Carbon 1 of pyruvate can be incorporated into malate and other citric acid cycle intermediates only via carboxylation, as this carbon would be lost via decarboxylation when pyruvate enters the citric acid cycle as acetyl-CoA via the pyruvate dehydrogenase reaction. The amount of malate formed equaled the 14CO2 formed and the radioactivity from C-1 of pyruvate recovered in malate slightly exceeded the formation of 14CO2 in agreement with our previous studies that reported a high rate of carboxylation of pyruvate in intact islets. When intact pancreatic islets were incubated with methyl [U-14C]succinate as a mitochondrial source of four-carbon dicarboxylic acids, radioactivity appeared in pyruvate and lactate. Taken together with previous studies, the current results suggest that during glucose-induced insulin secretion there is a shuttle operating across the mitochondrial membrane in which glucose-derived pyruvate is taken up by mitochondria and carboxylated to oxaloacetate by pyruvate carboxylase. The oxaloacetate is converted to malate which exits the mitochondrion, where, in the cytosol, it is decarboxylated to pyruvate in the reaction catalyzed by malic enzyme. This pyruvate re-enters mitochondrial pools. Such a cycle produces NADPH in the cytosol. Since it is a cycle, this shuttle can produce far more NADPH than the pentose phosphate pathway, which is known to be a very minor route of glucose metabolism in the islet.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Incorporation of isotopic carbon into cerebral glycogen from non-glucose substrates.

1. Measurable incorporation of radioactive carbon from [U-14C]pyruvate, [U-14C]-glutamate and [14C]bicarbonate into the glycogen synthesized by brain slices in vitro was demonstrated. 2. The fructose diphosphatase activity of guinea-pig brain was determined and found to be about 0.03 mumol of substrate degraded/min per g of fresh tissue. 3. The specific radioactivity of the glucose carbon from glycogen relative to that of the precursor added to the incubation medium gave approximate values of 0.195 for glucose, 0.006 for pyruvate, 0.039 for glutamate and 0.001 for bicarbonate.

Animals↗

Movement of carbon from vegetative cells to heterocysts in Anabaena cylindrica.

Radioactive carbon assimilated by vegetative cells of Anabaena cylindrica in the light passed via an intrafilamentous route into heterocysts in the dark. After several hours, label per heterocyst approximated label per vegetative cell. Much of the label entering heterocysts was not available for diffusional exchange back into vegetative cells.

Autoradiography↗

REPLACEMENT RATES FOR HUMAN TISSUE FROM ATMOSPHERIC RADIOCARBON.

Carbon-14, derived from the testing of thermonuclear weapons in the atmosphere of the Northern Hemisphere during 1961-62, has been found in human tissues including the brain in amounts which reflect the atmospheric concentration of carbon-14 as of several months earlier. In collagen of cartilage, the rate of uptake of carbon-14 is much slower than in other tissues; essentially no radioactive carbon was found in the collagen of 70-year-old adults that had been exposed to the comparatively high concentrations of carbon-14 in the atmosphere during the years 1954 to 1964. Individuals from the Southern Hemisphere show little increase in the carbon-14 content of their tissues at present, and detailed tests with individuals traveling to the Northern Hemisphere from the Southern allow closer scrutiny of the tissue replacement rates.

Adult↗

The formation of serine from glycine and formaldehyde by cell free extracts of Clostridium acidi-urici.

Cells of Clostridium acidi-urici which were grown in a medium containing uric acid were harvested, disrupted by sonication and centrifuged. After centrifugation the supernatant which served as the cell free extract was used to study the synthesis of serine from 2-14C glycine and formaldehyde. Serine was isolated from the reaction mixture by column chromatography. After identification by paper chromatography, serine was degraded carbon by carbon to locate the position of the labelled carbon. Radioactivity was confined almost exclusively to the alpha carbon of serine which was derived from the alpha carbon of glycine. Formaldehyde, therefore, binds at the alpha carbon of glycine to form serine. Conversion of serine to pyruvate was prevented by adding EDTA to the reaction mixture.

Cell-Free System↗

A simple technique for measuring relative renal blood flow.

To determine whether externally monitored early renal uptake of 131I-hippurate is proportional to renal blood flow, the renal uptake of 131I-hippurate at 1--2 min after injection was compared with the renal accumulation of radioactive carbonized microspheres in dogs. A renal artery catheter equipped with a balloon was used to decrease renal blood flow unilaterally. One minute after the intravenous injection of 100 muCi of 131I-hippurate, about 1 muCi of either 85Sr- or 98Nb-labeled carbon microspheres was injected into the left ventricle. Radioactivity was measured over both kidneys. The total radioactivity within each kidney region of interest was corrected for background and integrated over the 1--2 min after injection. Thirteen measurements of relative renal blood flow were made for seven dogs. The dogs were then killed and both kidneys were excised and counted for the radioactivity of the microspheres. The 1--2-min relative renal uptake of 131I-hippurate correlated well with relative microsphere uptake, suggesting that relative renal blood flow can be simply determined from the external measurement of renal uptake of 131I-hippurate.

Animals↗

Translocation of carbon-14 in sugarcane plants supplied with or deprived of phosphorus.

Translocation of radioactive carbon from the fed part of a blade to the rest of the plant was impaired by a deficiency in phosphorus only when the level of phosphorus was low enough to decrease growth.Phosphorus deficiency decreased the percentage of inorganic phosphorus more than that of organic phosphorus, indicating little effect of phosphorus deficiency upon phosphorylation.Thus phosphorus deficiency did not reduce phosphorylation enough to affect translocation. Evidence for the involvement of phosphorylation in translocation obtained from studies with potassium deficiency, light intensity and quality, 3-(p-chlorophenyl)-1, 1-dimethylurea and 3-(3,4-dichlorophenyl)-1, 1-dimethylurea, and carbon dioxide suggests that noncyclic photosynthetic phosphorylation provides the energy for phototranslocation.

Journal Article↗

Lipogenesis from U14C lactate in obese Zucker rat hepatocytes. Effect of albumin-bound oleate.

Lipogenesis from U(14C) lactate was studied in hepatocytes isolated from obese Zucker rats (fa/fa) their lean littermates (Fa/?) and Sprague Dawley rats. The distribution of radioactive carbon between the glycerol and the fatty acid moieties of the acylglycerols were studied. Radioactive lactate was better utilized for glycerol formation than it was for fatty acid formation in the obese rats. However, when oleate was added to the hepatocytic incubation medium, radioactive lactate was preferentially incorporated into the fatty acid moiety of the acylglycerols. Zucker obesity classified as a "metabolic obesity" by Meyer (1) depends upon abnormalities in carbohydrate metabolism associated with increased lipogenesis. This might be explained by biochemical shifts in the utilization of nutrients (2). Among the nutrients, lactate seems to be a better source of carbon than glucose for lipid synthesis (3). It has been shown that there is an increased hepatic portal blood concentration of lactate several hours after eating: about 4 mM in Wistar rats (4) and 10-15 mM in obese Zucker rats (3). We are interested in determinating the incorporation of carbon from lactate either into glycerol or into fatty acyl moieties of hepatic acylglycerols, and in determining the influence of exogenous fatty acids on acylglycerol synthesis, since a high level of circulating fatty acids in Zucker obese rats has been reported (5). Our purpose was to determine the incorporation of lactate into glycerol and fatty acyl moieties of acylglycerols, under the influence of oleate. Hepatocytes were isolated from ad libitum fed obese Zucker rats (fa/fa), their lean littermates (Fa/?) and Sprague-Dawley rats (SD). Incorporation of lactate was studied for three hours, in order to exclude short-term regulation effects and to allow oleate to be distributed into all cellular compartments.

Albumins↗

[High-yield synthesis of 14C labeled alicyclic amino acids with high specific radioactivity using potassium (14C) cyanide].

In order to study the tumor specificity of synthetic nonmetabolizing amino acids, 10 different 14C labeled alicyclic amino acids (3a-3j) were synthesized in high yield and with high specific radioactivity. Carbon-14 labeled alicyclic hydantoins (2a-2j) were synthesized from a small amount of radioactive potassium [14C] cyanide and corresponding ketones (1a-1j). Th 14C hydantoins (2a-2j) thus obtained were hydrolyzed without isolation to give 14C labeled alicyclic amino acids (3a-3j). The overall radiochemical yields of the amino acids (3a-3j) from potassium [14C] cyanide were 55.6-93.2% with radiochemical purity more than 99%. Specific activities of these 14C labeled compounds (3a-3j) were 209- 250 MBq/mmol (5.66-6.75 mCi/mmol). When non-radioactive potassium cyanide was not added as a carrier, 1-aminocyclopentane[14C]-carboxylic acid (3b) and 1-aminocyclooctane[14C]carboxylic acid (3e) were synthesized in the yield of 64.9 and 19.0% respectively with the specific activity exceed more than 1.85 GBq/mmol (50mCi/mmol).

Amino Acids↗

Following the path of carbon in photosynthesis: a personal story.

Chronological recognition of the intermediates and mechanisms involved in photosynthetic carbon dioxide fixation is delineated. Sam Ruben and Martin Kamen's development of application of radioactive carbon for the study of carbon dioxide fixation provided impetus and techniques for following the path of carbon in photosynthesis. Discovery The identity of the primary carboxylation enzyme and its identity with the major protein of photosynthetic tissues ('Fraction 1' protein of Sam Wildman) is reviewed. Memories are dimmed by sixty years of exciting discoveries exploration in newer fields [see Benson 2002 (Annu Rev Plant Biol 53: 1-25), for research and perspectives beyond the early Berkeley days].

Journal Article↗

Host-defense mechanisms in hidradenitis suppurativa.

Host-defense mechanisms were studied in seven patients with active hidradenitis suppurativa (HS). Granulocyte phagocytic function was measured by ingestion of Staphylococcus aureus labeled with radioactive carbon 14 and intracellular killing was determined by bactericidal pour plate method. Chemotaxis was measured by radioactive counting of sodium chromate Cr 51 granulocytes migrating in modified Boyden chambers. Granulocyte adherence was estimated in vitro by filtering blood samples through nylon fiber columns. Cell-mediated immunity was measured by intradermal delayed hypersensitivity responses to Candida, mumps. streptokinase/streptodornase, and purified protein derivative antigens. No abnormality was demonstrated in any granulocyte or cell-mediated immune function tests. Moreover, all patients had normal immunoglobulin levels and elevated total hemolytic complement. Therefore, we conclude that HS is a localized chronic infection of apocrine glands without a generalized defect in host defense.

Adult↗

Biosynthesis of caffeine in tea callus tissue.

1. A study of caffeine biosynthesis has been made by following the incorporation of radioactive carbon dioxide and methionine into the methylated purines produced by tea callus tissue. 2. The uptake of the radioactive labels into nucleic acid and caffeine was followed over a period of approximately 9h. 3. The distribution of the radioactive labels in both nucleic acid and caffeine was determined after incorporation and subsequent incubation of the tissue in a non-radioactive medium. 4. The results of the experiments indicated that the caffeine arose from purines released from the breakdown of nucleic acids rather than that it was formed directly from a purine pool. 5. A metabolic scheme to show the production of caffeine from the nucleotides of the nucleic acid is discussed.

Caffeine↗

Effect of Nitrogen Deficiency upon Translocation of C in Sugarcane.

Withholding nitrogen decreased the percentages of nitrogen and chlorophyll in the blades; reduced the total fixation of radioactive carbon dioxide at 15, 37, and 178 seconds; and changed the relative composition of fixation products. Translocation of radioactive photosynthate from the fed part down the attached blade and into the stalk was less in the plants deprived of nitrogen than in the control plants supplied with nitrogen. Both the percentage of total activity translocated and the velocity of transport were decreased by nitrogen deficiency. During a translocation period of 90 minutes the minus nitrogen blade retained more (14)C-sucrose than the control in the fed part and the blade below the fed part, but it sent less (14)C-sucrose to the sheath of the fed leaf. Thus translocation decreased with nitrogen deficiency not for lack of sucrose but for some other reason. Although withholding nitrogen decreased translocation of labeled carbon in and from attached blades, there was no effect upon transport in detached blades. The effect of nitrogen deficiency upon translocation may be indirect and secondary to the effect upon growth of the plant as a whole.

Journal Article↗