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Biomedical subjects

P C Shah

Publications and source records attributed to P C Shah.

At least 19 recordsLinked to original sources
PubMed2000

Genomic structure and expression of the murine Hmgi(y) gene.

Mammalian HMGI proteins belong to the high mobility group (HMG) of small non-histone nuclear proteins, and function as architectural factors to mediate structural changes in DNA. The HMGI family consists of three members: HMGI, HMGY and HMGI-C. As pseudogenes have complicated the genomic analysis of murine Hmgi(y), a mouse lambda FIX II genomic library was screened with an intron-specific probe to identify and characterize the authentic Hmgi(y) gene. The murine Hmgi(y) gene is 7.2kb long and contains four protein coding exons and two additional exons encoding part of the 5' untranslated region. Sequencing confirms that an alternative splicing site within exon 3 results in the two protein isoforms: Hmgi and Hmgy. Primer extension experiments revealed that at least three transcription start sites exist in the 5' end of the gene. It has been well established that the expression of both Hmgi-c and Hmgi(y) is readily detectable throughout embryogenesis. Unlike Hmgi-c, whose expression is restricted to embryogenesis, a Northern hybridization analysis showed low-level expression of Hmgi(y) in adult mouse tissues. Similarly, when tissues from newborn animals were examined, Hmgi(y) expression was readily detected at a level of intensity intermediate between that found in embryos and adults. Understanding the gene structure and expression pattern will provide important insights into the in-vivo function of Hmgi(y).

Amino Acid Sequence↗
PubMed2000

Activated recombinant human coagulation factor VII (rFVIIa) therapy for abdominal bleeding in patients with inhibitory antibodies to factor VIII.

Eight patients with inhibitors to factor VIII (4 hemophilia A and 4 nonhemophilic) were treated with recombinant activated factor VII (rFVIIa) to control severe abdominal bleeding. The recombinant factor was supplied under an open-label, emergency-use program to patients previously unresponsive to one or more alternative therapies. Therapy with rFVIIa was administered for nine separate bleeding events; one patient was treated for two separate bleeding episodes. Patients were treated for an average of 9 days and received a mean total dose of 5.2 mg of rFVIIa for control of bleeding. Treatment was considered successful and hemostasis adequate in 7 of the 9 episodes (78%). Treatment with rFVIIa was partially successful in one other episode. Four patients in this series experienced serious adverse events; all the adverse events were considered unrelated to rFVIIa therapy. The results of this limited series indicate that rFVIIa is an effective means of managing life-threatening abdominal bleeding in individuals with hemophilia or acquired antibodies to factor VIII.

Abdomen↗
PubMed2000

Human coagulation factor FVIIa (recombinant) in the management of limb-threatening bleeds unresponsive to alternative therapies: results from the NovoSeven emergency-use programme in patients with severe haemophilia or with acquired inhibitors.

This open-label, emergency-use study evaluated the efficacy and safety of activated human coagulation factor VIIa (recombinant) (rFVIIa) (NovoSeven; Novo Nordisk Pharmaceuticals, Inc., New Jersey, USA) in treating limb-threatening joint or muscle bleeds in 17 patients with haemophilia A or B and six patients with acquired inhibitors to factor VIII or factor IX. All patients had previously failed on one or more alternative therapies. rFVIIa administration was effective or partially effective in controlling joint or muscle bleeds in 34 out of 35 (97%) bleeding episodes; in 23 patients, 14 of 17 (82%) muscle bleeds and 16 of 18 (89%) joint bleeds were effectively controlled. These findings suggest that rFVIIa is an effective and well-tolerated therapeutic option in the management of joint or muscle haemorrhage in patients with haemophilia and in patients with acquired inhibitors.

Adolescent↗
PubMed1999

Effect of aging on intestinal ischemia and reperfusion injury.

Intestinal ischemia/reperfusion (I/R) is a serious disorder that is prevalent in elderly patients. Reactive oxygen species are implicated in the pathogenesis of intestinal I/R injury. Reactive oxygen species are also implicated in cellular senescence and aging. To test the hypothesis that aging exacerbates intestinal I/R injury, the effects of intestinal I/R on tissue injury were compared between young (3 month old) and aged (12 month old) mice. Intestinal ischemia was induced by occluding the superior mesenteric artery with a microbulldog clamp. Reperfusion was initiated by removing the clamp. Mortality due to intestinal ischemia followed by reperfusion was significantly higher in aged mice. There were no differences in the baseline levels of malondialdehyde or myeloperoxidase activity (indicators of lipid peroxidation and neutrophil infiltration, respectively) between young and aged mice. Although intestinal I/R caused a significant increase in malondialdehyde levels and myeloperoxidase activity in aged mice, similar increases were also observed in young mice. There were no significant differences in the activities of antioxidant enzymes including superoxide dismutase, glutathione peroxidase and catalase between young and aged mice that underwent sham operation. Intestinal I/R caused a significant decrease in catalase activity only in aged mice. In conclusion, our results indicate that aged mice are more susceptible to mortality due to intestinal I/R and that an age-dependent decrease in catalase activity may contribute to the observed mortality.

Aging↗
PubMed1999

Human ovarian cancer, cell lines, and primary ascites cells express the human Mullerian inhibiting substance (MIS) type II receptor, bind, and are responsive to MIS.

Six human ovarian cancer cell lines and samples of ascites cells isolated from 27 patients with stage III or IV ovarian papillary serous cystadenocarcinoma were studied individually to test whether recombinant human Mullerian inhibiting substance (rhMIS) acts via its receptor. To do these experiments, we scaled up production of rhMIS and labeled it successfully with biotin for binding studies, cloned the human MIS type II receptor for mRNA detection, and raised antibodies to an extracellular domain peptide for protein detection. These probes were first tested on the human ovarian cancer cell lines and then applied to primary ovarian ascites cells. rhMIS inhibited colony growth of five of six cell lines that expressed the human MIS type II receptor mRNA by Northern analysis while not inhibiting receptor-negative COS cells. Flow cytometry performed on MIS-sensitive ovarian cancer cell lines demonstrated specific and saturable binding of rhMIS (Kd = 10.2 nM). Ascites cells from 15 of 27 or 56% of patients tested bound biotinylated MIS (MIS-biotin) and, of the 11 that grew in soft agarose, 9 of 11 or 82% showed statistically significant inhibition of colony formation. Of the 15 patients who bound biotinylated MIS, mRNA was available for analysis from 9, and 8 of 9 expressed MIS type II receptor mRNA by reverse transcription-PCR, showing a statistically significant correlation, compared with binding, by chi2 analysis (P = 0.025). Solid ovarian cancers were positive for the MIS type II receptor protein by immunohistochemical staining, which colocalized with staining for antibody to CA-125 (OC-125). Thus, the detection of the MIS type I receptor by flow cytometry may be a useful predictor of therapeutic response to MIS and may be a modality to rapidly choose patients with late-stage ovarian cancer for treatment with MIS.

Adult↗
PubMed1997

Using altered specificity Oct-1 and Oct-2 mutants to analyze the regulation of immunoglobulin gene transcription.

Oct-1 and Oct-2 represent the prototypical example of related transcription factors with identical DNA recognition properties. They bind functionally critical octamer elements found in diverse regulatory sequences. It has not been possible to determine directly if these factors are functionally redundant or selective when interacting with different regulatory sequences in the appropriate cell type. An equivalent pair of altered DNA-binding specificity mutants of Oct-1 and Oct-2 are used to examine their function from varied regulatory contexts in B cells. These factors function as redundant activators of immunoglobulin (Ig) gene promoters (Vkappa and VH) and a histone H2B promoter. The structural basis of redundancy resides in the highly conserved DNA-binding POU domain, because this domain of either protein can activate transcription from both Ig and H2B promoters. We find that the nature of a distal enhancer dictates the relative potency of Oct-1 versus Oct-2 bound to a promoter. Oct-1 preferentially stimulates transcription from a VH or Vkappa promoter in combination with enhancers from the IgH or Igkappa locus, respectively. In this context, the more potent action of Oct-1 is dependent on a region external to the POU domain. These results suggest that Oct-1 may be the critical regulator of Ig gene transcription during B cell development and provide an explanation for selective Ig isotype expression defects in Oct-2 and OCA-B null mice.

Animals↗
PubMed1997

Intestinal ischemia and reperfusion injury in transgenic mice overexpressing copper-zinc superoxide dismutase.

Superoxide dismutase (SOD) scavenges oxygen radicals that are implicated in the pathogenesis of intestinal ischemia-reperfusion injury. The effect of intestinal ischemia and reperfusion was investigated in transgenic mice overexpressing human Cu-Zn SOD. Ischemia was induced by occluding the superior mesenteric artery. Myeloperoxidase activity was determined as an index of neutrophil infiltration, and malondialdehyde levels were measured as an indicator of lipid peroxidation. Forty-five minutes of intestinal ischemia followed by 4 h of reperfusion caused an increase in intestinal levels of malondialdehyde in both nontransgenic and transgenic mice, but the concentration of malondialdehyde was significantly greater in nontransgenic mice. Intestinal ischemia-reperfusion also caused an increase in intestinal and pulmonary myeloperoxidase activity in nontransgenic and transgenic mice, but the transgenic mice had significantly lower levels of myeloperoxidase activity than nontransgenic mice. Transgenic mice had higher levels of intestinal SOD activity than nontransgenic mice. There were no significant differences in the catalase or glutathione peroxidase activities. In conclusion, our study demonstrates that the overexpression of SOD protects tissues from neutrophil infiltration and lipid peroxidation during intestinal ischemia-reperfusion.

Animals↗
PubMed1996

The immunophilin FKBP12 functions as a common inhibitor of the TGF beta family type I receptors.

The immunophilin FKBP12 is an evolutionarily conserved abundant protein; however, its physiological roles remain poorly defined. Here we report that FKBP12 is a common cytoplasmic interactor of TGF beta family type I receptors. FKBP12 binds to ligand-free TGF beta type I receptor, from which it is released upon a ligand-induced, type II receptor mediated phosphorylation of the type I receptor. Blocking FKBP12/type I receptor interaction with FK506 nonfunctional derivatives enhances the ligand activity, indicating that FKBP12 binding is inhibitory to the signaling pathways of the TGF beta family ligands. Overexpression of a myristylated FKBP12 in Mv1Lu cell specifically inhibits two separate pathways activated by TGF beta, and two point mutations on FKBP12 (G89P, I90K) abolish the inhibitory activity of FKBP12, suggesting that FKBP12 may dock a cytoplasmic protein to the type I receptors to inhibit TGF beta family mediated signaling.

Activin Receptors, Type I↗
PubMed1996

The p21(RAS) farnesyltransferase alpha subunit in TGF-beta and activin signaling.

The alpha subunit of p21(RAS) farnesyltransferase (FNTA), which is also shared by geranylgeranyltransferase, was isolated as a specific cytoplasmic interactor of the transforming growth factor-beta (TGF-beta) and activin type I receptors with the use of the yeast two-hybrid system. FNTA interacts specifically with ligand-free TGF-beta type l receptor but is phosphorylated and released upon ligand binding. Furthermore, the release is dependent on the kinase activity of the TGF-beta type II receptor. Thus, the growth inhibitory and differentiative pathways activated by TGF-beta and activin involve novel mechanisms of serine-threonine receptor phosphorylation-dependent release of cytoplasmic interactors and regulation of the activation of small G proteins, such as p21(RAS).

Activin Receptors↗
PubMed1996

Developmental expression of a candidate müllerian inhibiting substance type II receptor.

We have isolated a candidate Müllerian inhibiting substance (MIS) type II receptor complementary DNA from an embryonic rat urogenital ridge library and have studied its binding to MIS, its developmental pattern of expression and tissue distribution. By in situ hybridization with a full-length riboprobe, the receptor is expressed in the mesenchymal cells surrounding the Müllerian duct at embryonic days 14, 15, and 16 and in tubular and follicular structures of the rat fetal gonads. Expression of the messenger RNA was also seen in the granules cells and seminiferous tubules of pubertal gonads. Northern analysis revealed that the MIS type II receptor messenger RNA is highly expressed in embryonic, pubertal, and adult testes and ovaries, as well as in the gravid uterus. The timing of expression in the gonads of both sexes was also analyzed by Northern analyses that showed high levels of expression at the time of Müllerian duct regression, much lower levels neonatally and prepubertally and then increased expression again with sexual maturation. The tissue and developmental specificity of expression of this receptor, which make it likely that this is the functional MIS type II receptor, can be used to advantage in therapeutic targeting strategies and to decipher the function of MIS in the gonads.

Aging↗
PubMed1996

Müllerian inhibiting substance as a model for the transforming growth factor-beta family: development of new treatment strategies.

The study of ligand receptor interactions and receptor function often requires multifaceted experimental approaches. In the course of studying the function and mechanism of action of müllerian inhibiting substance (MIS), we have used a wide range of molecular and cellular techniques. These have led to the identification, cloning, and characterization of the MIS receptors and of other receptors for the transforming growth factor-beta (TGF beta) family. This article describes the use of polymerase chain reaction (PCR) cloning to isolate candidate receptor genes, transfection and flow cytometry to study ligand binding, nonhomologous recombination targeted gene disruption (knockout) to analyze receptor function, and yeast genetics to identify other proteins that interact with the receptor complex. Together these techniques have led to the development of therapeutics and therapeutic strategies that are ready for clinical application.

Anti-Mullerian Hormone↗
PubMed1995

Confirmation of spurious hypoxemia using continuous blood gas analysis in a patient with chronic myelogenous leukemia.

Patients with extreme leukocytosis or thrombocytosis who have hypoxemia on arterial blood gas analysis may demonstrate normal oxygen saturation using pulse oximetry. The most commonly invoked explanation for this phenomenon is oxygen consumption in the blood gas sample prior to analysis. However, others have challenged the premise that the hypoxemia is spurious. We describe a patient with extreme leukocytosis and hypoxemia in whom normoxia was confirmed by continuous blood gas analysis.

Adult↗
PubMed1994

The v-abl tyrosine kinase negatively regulates NF-kappa B/Rel factors and blocks kappa gene transcription in pre-B lymphocytes.

Transformation of B-lineage precursors by the Abelson murine leukemia virus appears to arrest development at the pre-B stage. Abelson-transformed pre-B cell lines generally retain transcriptionally inactive, unrearranged immunoglobulin kappa alleles. We demonstrate that nontransformed pre-B cells expanded from the mouse bone marrow efficiently transcribe unrearranged kappa alleles. In addition, they contain activated complexes of the NF-kappa B/Rel transcription factor family, in contrast with their Abelson-transformed counterparts. Using conditionally transformed pre-B cell lines, we show that the v-abl viral transforming protein, a tyrosine kinase, blocks germ-line kappa gene transcription and negatively regulates NF-kappa B/Rel activity. An active v-abl kinase specifically inhibits the NF-kappa B/Rel-dependent kappa intron enhancer, which is implicated in promoting both transcription and rearrangement of the kappa locus. v-abl inhibits the activated state of NF-kappa B/Rel complexes in a pre-B cell via a post-translational mechanism that results in increased stability of the inhibitory subunit I kappa B alpha. This analysis suggests a molecular pathway by which v-abl inhibits kappa locus transcription and rearrangement.

Animals↗
PubMed1993

Human aldehyde dehydrogenase. cDNA cloning and primary structure of the enzyme that catalyzes dehydrogenation of 4-aminobutyraldehyde.

Human liver aldehyde dehydrogenase (E3 isozyme), with wide substrate specificity and low Km for 4-aminobutyraldehyde, was only recently characterized [Kurys, G., Ambroziak, W. & Pietruszko, R. (1989) J. Biol. Chem. 264, 4715-4721] and in this study we report on its primary structure. Polyclonal antibodies, specific for the E3 isozyme and three oligonucleotide probes derived from amino acid sequence of the E3 protein, were used for isolation of the first cDNA clone encoding the human enzyme (1503 bp; coding for 440 amino acid residues). Additional clones were obtained by using the first isolated clone as a probe. The largest clone of 1635 bp coded for 462 amino acid residues; it was longer at the 3'end of the cDNA non-coding region. The identity of the clone was established by DNA sequencing and by comparison with peptide sequences derived from the E3 protein, which constituted approximately 29% of the total primary structure of the E3 isozyme. The start codon was never encountered despite a variety of different approaches (500 amino acid residues were expected on the basis of SDS-gel molecular-mass determination of the E3 isozyme subunit). Despite the great catalytic similarity between the E3 and E1 isozymes [Ambroziak, W. & Pietruszko, R. (1991) J. Biol. Chem. 266, 13011-13018], the primary structure of the E3 isozyme has only approximately 40.6% of positional identity with that of the E1 isozyme. Sequence comparison with GenBank and Protein Identification Resource database sequences indicated no primary structure of aldehyde dehydrogenase more closely resembling the E3 isozyme than that of Escherichia coli betaine aldehyde dehydrogenase (52.7% positional identity), a prokaryotic enzyme specific for betaine aldehyde.

Aldehyde Dehydrogenase↗
PubMed1991

Locally recurrent endometrioid adenocarcinoma of the prostate after radical prostatectomy.

Endometrioid carcinoma of the prostate is considered a variant of classical prostatic ductal carcinoma. Endometrioid carcinoma variant often has the unique clinical presentation of gross hematuria. The propensity of this tumor to spread within the urothelium makes local failure of curative therapy commonplace. We present 2 representative cases with a review of followup surveillance procedures and treatment options for the local recurrence once identified.

Adenocarcinoma↗
PubMed1988

Tinea corporis caused by Microsporum canis: report of a nosocomial outbreak.

In an unusual nosocomial outbreak, 13 staff and 11 patients in an acute and chronic health care facility were infected with the zoophilic dermatophyte, Microsporum canis. The dermatophyte was apparently introduced into the facility by a single infected patient. Likely modes of subsequent disease transmission include person-to-person contact, handling of contaminated laundry, and use of a shared razor. Infection control measures for managing such outbreaks are discussed.

Adult↗