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Biomedical subjects

D D Rao

Publications and source records attributed to D D Rao.

16 recordsLinked to original sources

Intestinal distribution of human Na+/H+ exchanger isoforms NHE-1, NHE-2, and NHE-3 mRNA.

The identity of Na+/H+ exchanger (NHE) isoforms in the human small intestine and colon and their role in vectorial Na+ absorption are not known. The present studies were undertaken to examine the regional and vertical axis distribution of NHE-1, NHE-2, and NHE-3 mRNA in the human intestine. Ribonuclease protection assays were used to quantitate the levels of mRNA of these isoforms in various regions of the human intestine. In situ hybridization technique was used to localize NHE-2 and NHE-3 mRNA in the colon. The NHE-1 isoform message was present uniformly throughout the length of the human intestine. In contrast, mRNA levels for human NHE-2 and NHE-3 isoforms demonstrated significant regional differences. The NHE-3 abundance was found in decreasing order: ileum > jejunum > proximal colon = distal colon. The NHE-2 message level in the distal colon was significantly higher than in the proximal colon but was evenly distributed in the small intestine. In addition, NHE-2 mRNA was present in surface epithelial cells as well as in cells of the crypt region, suggesting the presence of NHE-2 message throughout the vertical axis of the colonic crypts. In contrast, NHE-3 mRNA was localized to surface colonocytes in the proximal colon. On the basis of this tissue-specific localization of NHE-2 and NHE-3 mRNA, it can be speculated that the relative contribution of NHE-2 and NHE-3 isoforms in Na+ absorption in the human intestine may be region specific, and these putative apical isoforms may be differentially regulated.

Amino Acid Sequence↗

Rodent and human beta 3-adrenergic receptor genes contain an intron within the protein-coding block.

DNA blot analysis of the cloned rat beta 3-adrenergic receptor gene revealed unexpected restriction enzyme cleavage sites that suggested the presence of one or more introns near the end of the coding block. This region of the rat gene was mapped and sequenced and was found to contain two introns. The first intron occurs 12 amino acids from the end of the coding block, as deduced by comparison with the beta 3 receptor cDNA. Sequence analysis of the first intron indicates that it might contain enhancer elements that could be important in the adipose tissue-specific expression of this gene. The mouse and human beta 3 receptor genes have been assumed to be intronless; however, these genes contain potential splice sites that are homologous to those present in the rat gene. The relevant regions of the mouse and human beta 3 receptor cDNAs were cloned and, by comparing them to the respective genomic sequences, it was concluded that these genes also contain one or more introns. Sequence analysis of the mouse and human beta 3 receptor cDNAs indicates that they code for proteins that are, respectively, 12 and 6 amino acids larger than previously deduced from genomic clones.

Adipose Tissue↗

Two forms of the rat D2 dopamine receptor as revealed by the polymerase chain reaction.

We have used the polymerase chain reaction technique (PCR) to clone the cDNA of the D2 dopamine receptor from rat striatal mRNA. Two major PCR products were produced; one product was identical to a previously published rat cDNA, while the other, more abundant product differed only by an 87-nucleotide insert located in the region of the putative third cytoplasmic loop of the D2 receptor. A PCR approach for determining message abundance was used to determine the relative message abundance of the two forms of the D2 receptor in a variety of tissues. Possible implications of the two forms of the D2 receptor for dopamine-mediated signal transduction are discussed.

Amino Acid Sequence↗

Vitamin E and relationships among tocopherols in human plasma, platelets, lymphocytes, and red blood cells.

Plasma, RBC, platelets, and lymphocytes from human subjects on graded intakes of vitamin E were analyzed for tocopherols to determine which humoral compartment most closely followed changes in the dietary intake. Relative merits of the various blood elements to reflect changes in vitamin E intake were calculated by the sensitivity concept of Mandel and Stiehler (ie, rate of change of tocopherol levels with dose divided by the standard deviation). Sensitivities of alpha-tocopherol levels of the various blood components to vitamin E intake decreased in the order platelets greater than RBC greater than plasma lipids greater than plasma greater than lymphocytes. Changes in tocopherol levels in platelets most closely followed changing dietary intakes of vitamin E when compared with data from RBC, lymphocytes, and plasma. Furthermore, for studying the effects of diets on vitamin E status, tocopherol levels of platelets appear to be a better measure than tocopherol levels of either RBC, lymphocytes, or plasma lipids.

Blood Platelets↗

Interference among defective interfering particles of vesicular stomatitis virus.

Three defective interfering (DI) particles of vesicular stomatitis virus (VSV), all derived from the same parental standard San Juan strain (Indiana serotype), were used in various combinations to infect cells together with the parental virus. The replication of their RNA genomes in the presence of other competing genomes was described by the hierarchical sequence: DI 0.52 particles greater than DI 0.45 particles less than or equal to DI-T particles greater than standard VSV. The advantage of one DI particle over another was not due simply to multiplicity effects nor to the irreversible occupation of limited cellular sites. Interference, however, did correlate with a change in the ratio of plus and minus RNA templates that accumulated intracellularly and with the presence of new sequences at the 3' end of the DI genomes. DI 0.52 particles contained significantly more nucleotides at the 3' end that were complementary to those at the 5' end of its RNA than did DI-T or DI 0.45 particles. The first 45 nucleotides at the 3' ends of all of the DI RNAs were identical. VSV and its DI particles can be separated into three classes, depending on their terminal RNA sequences. These sequences suggest two mechanisms, one based on the affinity of polymerase binding and the other on the affinity of N-protein binding, that may account for interference by DI particles against standard VSV and among DI particles themselves.

Animals↗

RNA synthesis of vesicular stomatitis virus. X. Transcription and replication by defective interfering particles.

In cells coinfected by standard vesicular stomatitis virus (VSV) and defective interfering (DI) T particles, small RNA consisting of 46 nucleotides was synthesized in molar excess over other VSV-specific RNAs. Although its rate of synthesis increased over time, small RNA accumulated linearly, suggesting that the molecule is unstable. In contrast, replication of the genome RNA of DI T particles was relatively constant after 3 h of infection, resulting in the intracellular accumulation of stable genomic and antigenomic RNA of DI T particles. Coinfection of cells with DI T particles and selected temperature-sensitive mutants from all five complementation groups of VSV indicated that the replication of DI genomes was controlled separately from the synthesis of small RNA. Also, when viral RNA replication was inhibited by cycloheximide, small RNA continued to be synthesized as long as there were enough templates present. These results indicate that small RNA is synthesized by the enzyme(s) involved in VSV transcription and that its dependence on RNA replication is due to the requirement for template amplification.

Animals↗

RNA synthesis of vesicular stomatitis virus. VIII. Oligonucleotides of the structural genes and mRNA.

The single-stranded RNA genome of vesicular stomatitis virus (VSV, Indiana serotype, San Juan strain) yields approx. 75 RNase T1-resistant oligonucleotides ranging in size from 10 to 50 bases. Each of the five structural genes, isolated as duplex RNA molecules hybridized to complementary mRNA, contains two or more of these large oligonucleotides. One of the oligonucleotides is identified as part of the non-coding region near the 3' end of the genome. Comparison of these results with others indicate that the RNA sequence of VSV is apparently stable in the laboratory but not in the wild. RNase T1-resistant oligonucleotides are also shown for all five VSV mRN species. Whether the mRNA for these digestions are are isolated from duplex RNA molecules or as single-stranded RNA species, the oligonucleotide patterns for each mRNA are virtually identical, indicating that each mRNA is transcribed from contiguous sequences on the genome. Comparison with published oligonucleotide patterns obtained from other isolates of VSV or from VSV deletion mutants indicate that identity and changes in their genome structure can be correlated with specific structural genes.

Base Sequence↗

Synthesis of a small RNA in cells coinfected by standard and defective interfering particles of vesicular stomatitis virus.

A small RNA, containing approximately 50 nucleotides, is synthesized by cells coinfected with standard vesicular stomatitis virus and its defective interfering (DI) particles. Infection of cells by standard virus or DI particles alone does not lead to synthesis of significant amounts of small RNA. The RNA is initiated at its 5' end with (p)ppXp and is not polyadenylylated at the 3' end despite a content of 51% adenosine. It has sequences complementary to the genome of a DI particle. The synthesis of the small RNA correlates with the replication of the genome of DI particles with molar ratio small RNA/genome RNA of DI particles greater than 50. When replication of DI genomes is prevented by the addition of cycloheximide or prior UV irradiation of DI particles, small RNA is not synthesized in coinfected cells. These results indicate that the small RNA is not the result of transcriptional initiation and that it may relate to interference mediated by DI particles.

Animals↗

Anorexia nervosa with acute tubular necrosis treated with parenteral nutrition.

A patient with nonoliguric acute renal failure secondary to acute tubular necrosis in conjunction with anorexia nervosa is described. Parenteral feeding at a critical time has salutory effects on the biosynthesis of new protein and thereby reduces many of the hazards of azotemia. The technique of estimating endogenous acid production is applied for the first time in a severely malnourished subject and documents the retention of dietary sulfur which presumably is retained in the formation of new tissue in the recovery phase.

Acute Kidney Injury↗

Renal tubular acidosis: practical guides to diagnosis and treatment.

The syndrome of renal tubular acidosis in some one of its various forms should be suspected when an infant or child has failure to thrive, metabolic acidosis, constipation, diarrhea, vomiting, anorexia, polyuria, or dehydration in infancy. Confirmatory biochemical findings include an inappropriately high urinary pH, inadequate acid excretion and/or abnormal tubular reabsorption of filtered bicarbonate. Growth can be normal when there is sustained correction of the metabolic acidosis through appropriate alkaline therapy.

Acidosis, Renal Tubular↗

A study on cancer mortality in Tarapur-based atomic energy community.

Cancer mortality risks for individuals who were employed at nuclear facilities in Tarapur and for their respective family members with whom they lived were examined. Cancer deaths that occurred in this population between 1971 and 1988 were compared with death rates published by the Bombay City Cancer Registry. Risks were expressed as standardized mortality ratios (SMRs), which were computed by dividing the observed number of deaths by the expected number of deaths and multiplying this value times 100. There were 11 deaths from cancer among the employees, and this figure was too small to permit any trend analysis with respect to radiation exposures. The SMRs for all cancers and leukemia for male employees and for middle-aged male family members who were not exposed to any radiation were not statistically significant. A much larger database of person years at risk would be required to reach definite conclusions. The combined cancer risks for employees and families combined were similar to risks experienced by individuals in Bombay.

Adult↗