Biomedical subjects

R Reddy

Publications and source records attributed to R Reddy.

At least 73 records · Page 4Linked to original sources

PubMed1999

Odorants induce the phosphorylation of the cAMP response element binding protein in olfactory receptor neurons.

Although odorants are known to activate olfactory receptor neurons through cAMP, the long-term effects of odorant detection are not known. Our recent findings indicate that there is also a delayed and sustained cAMP response, with kinetics sufficient to mediate long-term cellular responses. This cAMP response is mediated by cGMP through activation of adenylyl cyclase by protein kinase G (PKG). Therefore, we investigated the ability of odorants to regulate gene expression in rat olfactory epithelium. The cAMP-responsive binding protein (CREB) is a well-characterized transcription factor regulated by cAMP. We examined CREB activity in rat olfactory epithelium and olfactory receptor neurons (ORNs) after stimulation with odorants. Odorants increased levels of phosphorylated CREB in olfactory epithelium in vivo, and this increase was localized to ORNs in vitro. Incubation with 8-bromo-cGMP or sodium nitroprusside, a guanylyl cyclase activator, also increased phosphorylated CREB. In vitro, cAMP-dependent protein kinase phosphorylated CREB. In contrast, PKG failed to phosphorylate CREB directly in vitro. Our results demonstrate that the delayed odorant-induced cAMP signal activates CREB, which in turn may modulate gene expression in ORNs. In addition, cGMP indirectly affects CREB activation. This effect of cGMP on CREB activity through cAMP provides another mechanism for the modulation of CREB.

Animals↗

PubMed1999

Post-transcriptional adenylation of signal recognition particle RNA is carried out by an enzyme different from mRNA Poly(A) polymerase.

A fraction of the signal recognition particle (SRP) RNA from human, rat, Xenopus, and Saccharomyces cerevisiae cells contains a single post-transcriptionally added adenylic acid residue on its 3'-end; in the case of human SRP RNA, over 60% of the SRP RNA molecules contain a nontemplated adenylic acid residue on their 3'-ends (Sinha, K. M., Gu, J., Chen, Y., and Reddy, R. (1998) J. Biol. Chem. 273, 6853-6859). In this study, we investigated the enzyme that is involved in this 3'-end adenylation of SRP RNA. A U1A protein peptide conjugated to albumin completely inhibited the polyadenylation of a SV40 mRNA by HeLa cell nuclear extract in vitro; however, the 3'-end adenylation of human SRP RNA or Alu RNA, which corresponds to 5' and 3'-ends of SRP RNA, was not affected by this U1A peptide conjugate. SRP RNA from mutant strains of S. cerevisiae with a temperature-sensitive mRNA poly(A) polymerase grown at a restrictive temperature of 37 degrees C also contained a post-transcriptionally added adenylic acid residue just like SRP RNA from wild-type cells and mutant cells grown at permissive temperature of 23 degrees C. In addition, binding of SRP 9/14-kDa protein heterodimer was required for adenylation of Alu RNA in vitro. These lines of evidence, along with other data, show that post-transcriptional adenylation of SRP and Alu RNAs is carried out by a novel enzyme that is distinct from the mRNA poly(A) polymerase, CCA-adding enzyme, and nonspecific terminal transferase.

Alu Elements↗

PubMed1999

Detection of residual quadrupolar interaction in the human breast in vivo using sodium-23 multiple quantum spectroscopy.

Sodium multiple quantum (MQ) spectroscopy of the human breast in vivo was performed. Double quantum (DQ) filtered spectra were used to demonstrate the existence of a non-vanishing (residual) quadrupolar interaction in the tissue. Triple quantum (TQ) filtered spectra were used to measure the two time constants associated with the biexponential transverse relaxation times of sodium in biological tissues. The two time constants were found to be 0.64 and 26.57 msec. The potential applications of this finding are discussed.

Anisotropy↗

PubMed1999

A novel approach to observing articular cartilage deformation in vitro via magnetic resonance imaging.

The design of a pressure cell that compresses a cartilage specimen in one dimension within an imaging magnet is presented. One-dimensional projection images in a direction perpendicular to the articular surface of the cartilage specimen were used to generate a uniaxial confined deformation creep curve for normal and trypsin-degraded cartilage specimens during a continuous 0.690 MPa (100 psi) pressure application. The resulting curves are shown to fit a two time constant viscoelastic model well and also indicate that the elastic modulus of cartilage decreases and the deformation rate increases upon trypsin proteolysis. Furthermore, cartilage permeability is shown as a function of cartilage strain for both the normal and trypsin-degraded case. Several two-dimensional slice-selective images were collected both before and after 80 minutes of continuous compression. These images were used to evaluate the relative changes in the spin-lattice, T1, and spin-spin, T2, relaxation time constant maps for both normal and degraded cartilage specimens in response to compression. The results of this study demonstrate the utility of a novel, non-magnetic, cartilage compression device and also support the validity of a simple two-component rheological model of articular cartilage.

Animals↗

PubMed1999

Sodium and proton MR properties of cartilage during compression.

Proton and sodium MR relaxation times of bovine articular cartilage specimens were measured as a function of proteoglycan (PG) depletion and as a function of mechanical compression. Proton and sodium relaxation times of normal cartilage were compared with relaxation times of PG-depleted cartilage to evaluate the significance of PG depletion-induced changes in MR relaxation parameters. These comparisons were conducted for both uncompressed and mechanically compressed states. The mechanical compressions were performed with an MR-compatible pressure cell and evaluated dynamically via interleaved one-dimensional proton and sodium MR projection imaging. The comparisons indicate that sodium relaxation parameters are sensitive to PG depletion when cartilage is in a mechanically compressed state or an uncompressed state. In contrast, proton relaxation parameters do not change significantly with PG depletion when cartilage is in an uncompressed state. However, during mechanical compression, proton T2 becomes sensitive to PG depletion. These results support the potential of sodium magnetic resonance imaging (MRI) as a possible modality for obtaining imaging contrast related to PG depletion. The results also indicate the potential of proton MRI to provide such contrast if the image acquisition is conducted in conjunction with a mechanical compression via physical exercise.J. Magn. Reson Imaging 10:961-967, 1999.

Animals↗

PubMed1999

Sodium NMR evaluation of articular cartilage degradation.

One of the first effects of degenerative osteoarthritis is the loss of proteoglycans from the matrix of articular cartilage. Using a model of osteoarthritic change where the cartilage has been enzymatically degraded with trypsin, the sodium NMR characteristics of the cartilage were determined as a function of changes in the proteoglycan content. The results demonstrate that the single quantum sodium signal decreases as the proteoglycan content of the cartilage matrix decreases. In addition, the relaxation characteristics of the sodium change such that the T1 is longer, the T2s is longer, and the T2f is shorter. Short echo-time, T1-weighted sodium images are presented which demonstrate that this information may be utilized to detect the loss of proteoglycans from articular cartilage.

Animals↗

PubMed1999

In vivo triple quantum filtered twisted projection sodium MRI of human articular cartilage.

In this work, we present the first triple quantum filtered (TQF) sodium MR images of the human knee joint in vivo. A 3D TQF data set of 16 slices was obtained in 20 min using a TQF pulse sequence preencoded to a twisted projection imaging readout. Images clearly demarcate patellar cartilage and also demonstrate fluid signal suppressed by the triple quantum filter. Biexponential transverse relaxation times were calculated by fitting the TQF free induction decay to a theoretical signal expression. The average values from three healthy volunteers were T(2fall)(*) = 9.59 +/- 0.35 ms and T(2rise)(*) = 0.84 +/- 0.06 ms. Application of TQF imaging in biological tissues is discussed.

Cartilage, Articular↗

PubMed1999

Acute myo-inositol enhances swimming activity in goldfish (short communication).

Inositol in concentrations of 1-4 Mm (but not the control condition: mannitol-glucose) administered in aquarium water showed enhancement of swimming activity of goldfish after acute treatment (5 hours). These data support similar findings in rats. These data suggest that inositol may enhance motor activity which may be of relevance in neuropsychiatric disorders. Future studies of the effects of lithium on this activity, and its potential reversal by inositol may shed light on the possible involvement of the phosphatidyl-inositol second messenger system in this behavior.

Analysis of Variance↗

PubMed1999

An MR imaging method for simultaneous measurement of gaseous diffusion constant and longitudinal relaxation time.

A magnetic resonance imaging method for simultaneous and accurate determination of gaseous diffusion constant and longitudinal relaxation time is presented. The method is based on direct observation of diffusive motion. Initially, a slice-selective saturation of helium-3 (3He) spins was performed on a 3He/O2 phantom (9 atm/2 atm). A time-delay interval was introduced after saturation, allowing spins to diffuse in and out of the labeled slice. Following the delay interval a one-dimensional (1-D) projection image of the phantom was acquired. A series of 21 images was collected, each subsequent image having been acquired with an increased delay interval. Gradual spreading of the slice boundaries due to diffusion was thus observed. The projection profiles were fit to a solution of the Bloch equation corrected for diffusive motion. The fitting procedure yielded a value of D3He = 0.1562+/-0.0013 cm2/s, in good agreement with a measurement obtained with a modified version of the standard pulsed-field gradient technique. The method also enabled us to accurately measure the longitudinal relaxation of 3He spins by fitting the change of the total area under the projection profiles to an exponential. A value of T1 = 1.67 s (2 T field) was recorded, in excellent agreement with an inversion recovery measurement.

Diffusion↗

PubMed1999

Metronidazole and clarithromycin resistance amongst Helicobacter pylori isolates from a large metropolitan hospital in the United States.

BACKGROUND: Metronidazole and clarithromycin-based therapies are among the most efficacious treatment regimens for H. pylori infection. Resistance to metronidazole or clarithromycin is associated with impaired therapy with these agents. We conducted a retrospective review of susceptibility data to determine the frequency of primary metronidazole and clarithromycin resistance among H. pylori isolates from a single metropolitan hospital in the United States. The database comprised 933 patients who presented at the Digestive Diseases Clinic at the Veterans Affairs Medical Center in Houston between September 1988 and January 1997 with complaints of dyspepsia, abdominal pain and peptic ulcer disease. One hundred and seventy-nine of these patients had both pharmaceutical records available for evaluation and culture and antimicrobial susceptibility data for analysis. The MICs were determined by both E-test and broth microdilution tests. The frequency of primary metronidazole resistance was 37.4% (67/179). The level of primary clarithromycin resistance was 6.1%. Dual metronidazole and clarithromycin resistance was present in approx. 3%. The high level of primary metronidazole and clarithromycin resistance in H. pylori isolates from this metropolitan hospital is such that antimicrobial susceptibility data should be available so that informed choice can be made for specific eradication therapies, especially in patients who fail treatment.

Anti-Bacterial Agents↗

PubMed1999

An RNA polymerase III transcription unit located in the upstream control regions of the human proliferating-cell nucleolar protein p120 gene is transcribed in vitro and in vivo.

An RNA polymerase III (Pol III) transcription unit containing homology to highly repeated Alu sequences was identified in the upstream flanking sequences of the gene for the human proliferating-cell nucleolar antigen p120. When transcribed in vitro, this Pol III unit produced three RNA transcripts, designated by nucleotide length as T150, T385 and T635; RNA transcript T635 was the most abundant accounting for over 90%. The transcription initiates at nucleotide -729 of the human p120 promoter and proceeds in the opposite orientation to the p120 gene transcription. Northern blot analysis and cDNA cloning followed by sequencing showed the presence of the T635 RNA in HeLa cells, indicating that this Pol III transcription unit is functional and transcribed in vivo. Disruption of this Pol III transcription unit by deletion of the Box A residues (-733 to -744) resulted in a sixfold reduction of the p120 gene transcription. A possible role for this Pol III transcription unit in p120 gene transcription is discussed.

Base Sequence↗

PubMed1999

Bcl-2 and p53 immunoprofile in Kaposi's sarcoma.

Seventy three cases of Kaposi's sarcoma (KS) from the 3 histological subtypes (patch, plaque and nodular) were assessed for bcl-2 and p53 protein expression. The aim was to determine the level of expression of these proteins in KS and in the different subtypes. Commercially available antibodies to bcl-2 and p53 were applied after both microwave and pressure cooking antigen retrieval. Bcl-2 immunoexpression increased from the patch stage (36%) to the plaque stage (45%) to the nodular stage (70.83%). Better immunostaining for bcl-2 was obtained after pressure cooking. p53 on the other hand, was not expressed in the patch or plaque stages, but 54.16% of cases in the nodular stage were immunopositive. These results show a progression of immunoexpression of both bcl-2 and p53 from the early histological stages to the late tumor stage, implying that these proteins are upregulated late in the evolution of KS.

Acquired Immunodeficiency Syndrome↗

PubMed1999

Hepatitis C virus genotypes and viremia and hepatocellular carcinoma in the United States.

OBJECTIVE: Hepatitis C virus (HCV) is a well recognized cause of hepatocellular carcinoma (HCC). The pathogenic significance of HCV genotypes in hepatocarcinogenesis is undefined. The aim of this study was to investigate the genotypic distribution and viremic level of HCV in patients with HCV-associated cirrhosis with or without HCC. METHODS: A total of 28 HCV-infected patients with HCC (HCC+) and 38 patients with HCV-associated cirrhosis without HCC (HCC-) were studied. HCV genotype was assessed by the genotype-specific polymerase chain reaction (PCR) method of Okamoto and restriction fragment length polymorphism (RFLP) of the 5' untranslated region (5' UTR). Hepatitis C viremia was quantitated with the branched-chain DNA (bDNA) assay. RESULTS: Using the Okamoto method, we found genotype 1b in 64% of the HCC+ group and 74% of the HCC- group, 36% of the HCC+ group and 16% of the HCC- group were coinfected with a combination of genotype 1b and another genotype. Using the RFLP method, we found genotype 1b in 41% of the HCC+ group and in 24% of the HCC- group. Other genotypes accounted for 18% of the HCC+ group and 55% of the HCC- group; no combination genotypes were identified. Poor concordance occurred between the two genotyping methods. Mean bDNA levels were not significantly different between the two groups. CONCLUSIONS: Our study demonstrates that no particular HCV genotypes were associated with HCC and genotype did not appear to influence the development of HCV-associated HCC.

Carcinoma, Hepatocellular↗

PubMed1999

Pseudo-outbreak of methicillin-resistant Staphylococcus aureus.

OBJECTIVE: To determine whether a high rate of methicillin-resistant Staphylococcus aureus at our institution was due to laboratory misclassification and to evaluate the effect of this misclassification. MATERIAL AND METHODS: We evaluated all S. aureus isolates identified at our institution during a 60-day period in 1997. Automated susceptibility test results (using the Vitek system) from our clinical microbiology laboratory and an independent laboratory were compared with oxacillin agar screen plate results at both laboratories. Isolates with discordant results for susceptibility to oxacillin were tested by broth microdilution minimal inhibitory concentrations and for the presence of the mecA gene. RESULTS: Eighteen (72%) of the 25 organisms (obtained from 17 patients) found to be resistant to oxacillin by the Vitek system at our institution were susceptible by the oxacillin agar screen. Discordant isolates tested by broth microdilution minimal inhibitory concentrations and for the mecA gene were found to be oxacillin susceptible and mecA gene negative. Thus, at our hospital, almost three fourths of the organisms initially identified as methicillin-resistant S. aureus by the Vitek system were actually susceptible to oxacillin. This misclassification resulted in needless infection control measures and unnecessary vancomycin use. CONCLUSION: Hospitals that use only automated susceptibility testing for S. aureus should periodically validate their results with additional testing.

Cross Infection↗

PubMed1998

Accurate 3' end processing and adenylation of human signal recognition particle RNA and alu RNA in vitro.

Human signal recognition particle (SRP) RNA is transcribed by RNA polymerase III and terminates with -GUCUCUUUUOH on its 3' end. Our previous studies showed that the three terminal uridylic acid residues of human SRP RNA are post-transcriptionally removed and a single adenylic acid residue is added, resulting in a 3' end sequence of -GUCUCUAOH (Sinha, K. M., Gu, J., Chen, Y., and Reddy, R. (1998) J. Biol. Chem. 273, 6853-6859). In this study we show that the Alu RNA, corresponding to the 5' and 3' ends of SRP RNA, is also accurately processed and adenylated in vitro. Alu RNAs containing 7 or 11 additional nucleotides on the 3' end were accurately processed and then adenylated. Deletion analysis showed that an 87-nucleotide-long motif comprising of the 5' and 3' ends, including stem IV of the Alu RNA, is sufficient and necessary for the 3' end processing and adenylation. A 73-nucleotide-long construct with deletion of stem IV, required for the binding of SRP 9/14-kDa proteins, was neither processed nor adenylated. The adenylated Alu RNA as well as adenylated SRP RNA were bound to the SRP 9/14-kDa heterodimer and were immunoprecipitated by specific antibodies. A significant fraction of SRP RNA in the nucleoli was found to be processed and adenylated. These data are consistent with nascent SRP and/or Alu RNAs first binding to SRP 9/14-kDa protein heterodimer, followed by the removal of extra sequence on the 3' end and then the addition of one adenylic acid residue in the nucleus, before transport into the cytoplasm.

Adenosine Monophosphate↗

PubMed1998

In vivo expression of a variant human U6 RNA from a unique, internal promoter.

We previously isolated a variant of the human U6 small nuclear RNA gene (87U6) and demonstrated that transcription of this gene is controlled by a novel internal promoter. It has now been shown that two blocks of sequence within the coding region are both necessary and sufficient to direct expression of 87U6 in transcription assays performed in vitro. In addition, 87U6 is expressed in vivo and can assemble into snRNP complexes. Specific primer extension assays on total RNA from HeLa cells shows that 87U6 RNA is present in these cells. Also, microinjection of plasmid encoded 87U6 genes into Xenopus laevis oocyte nuclei results in the expression of this variant RNA. Immunoprecipitation with anti-Sm antibodies suggests that 87U6 RNA assembles into a snRNP particle with U4 snRNA. Finally, the variant snRNA is capped with the U6 specific gamma-monomethyl phosphate cap when incubated in HeLa extracts. These data suggest that 87U6 RNA may function in the splicing process, in a manner similar to the wild-type U6 RNA. The recent observations of a minor class of mRNA introns that are spliced by a distinct collection of snRNP particles suggest an important role for variant snRNAs in the splicing of transcripts with alternative splice junctions.

Animals↗

PubMed1998

Isolation and characterization of a new 110 kDa human nuclear RNA-binding protein (p110nrb).

RNA-protein interactions play key roles in many fundamental cellular processes such as RNA processing, RNA transport, and RNA translation. During our attempts to isolate the human U6 small nuclear RNA capping enzyme, we identified a new 110 kDa nuclear RNA-binding protein, designated p110nrb. The full-length cDNA clone for p110nrb was characterized, and it encodes a 963 amino acid polypeptide. It is a highly acidic protein (pI 5.28) and the carboxyl terminal portion contains two conserved RNP motifs. A databank search found a putative C. elegans protein that might be the p110nrb homologue. The p110nrb was overexpressed as a glutathione S-transferase fusion protein in insect Sf9 cells, purified by affinity chromatography and injected into rabbits to produce specific polyclonal antibodies. Immunofluorescent staining showed that p110nrb is distributed evenly throughout the nucleoplasm. Northern blots showed that the mRNA is expressed in all tissues examined. An in vitro RNA-binding assay showed that p110nrb bound to RNA. These data suggest that p110nrb may play a role in the metabolism of nuclear RNA.

Amino Acid Sequence↗

PubMed1998

Reduced level of plasma antioxidant uric acid in schizophrenia.

There is evidence of dysregulation of the antioxidant defense system in schizophrenia. The purpose of the present study was to examine whether uric acid, a potent antioxidant, is reduced in the plasma of patients with schizophrenia. To this end, a within-subject, repeated measures, on-off-on haloperidol treatment design was utilized. Male schizophrenic patients with either a haloperidol treatment (n=47) or a drug-free condition (n=35) had significantly lower levels of plasma uric acid than the age- and sex-matched normal control subjects (n=34). Following haloperidol withdrawal, plasma uric acid levels were further reduced in schizophrenic patients (P=0.018; paired t-test, n=35). However, no relationship was found between uric acid levels and the length of the drug-free period (< 5 or > 5 weeks) or days drug free. In addition, the plasma levels of uric acid in patient groups were significantly and inversely correlated with psychosis. There was a trend for lower uric acid levels in relapsed patients relative to clinically stable patients. Smoking, which can modify plasma antioxidant capacity, was not found to have prominent effects on uric acid levels. The present finding of a significant decrease of a selective antioxidant provides additional support to the hypothesis that oxidative stress in schizophrenia may be due to a defect in the antioxidant defense system.

Adult↗