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M Patel

Publications and source records attributed to M Patel.

At least 271 records · Page 15Linked to original sources
PubMed1995

The carboxy-terminal domain is essential for stability and not for virion incorporation of HIV-1 Vpr into virus particles.

Vpr is one of the auxiliary gene products encoded by HIV-1 genome. Vpr is a 14-kDa protein and exhibits several interesting characteristics including incorporation into virus particles, oligomerization, localization in the nucleus, and positive regulation of virus replication in primary cells. In an effort to define the structure-function relationship of Vpr, the role of the C-terminus of Vpr was investigated. Site-specific mutagenesis involving deletion, insertion, and substitution of residues at the C-terminus was utilized to generate variants of Vpr. Mutations introduced at the C-terminus affected properties of Vpr in different ways: (i) Vpr containing amino acids 1-72 showed the virion incorporation phenotype, indicating that the C-terminus is not essential for this function, (ii) the C-terminus contributes to the stability of Vpr, and (iii) substitution mutagenesis involving the basic residues showed stability similar to that of wild type, indicating the lack of involvement of these residues in this biochemical property of Vpr. The data generated in this study and our early mutagenic analyses on Vpr suggest that domains noncontiguous in primary sequence contribute to the stability of Vpr through overall conformation of the protein.

Amino Acid Sequence↗
PubMed1995

Functional analysis of HIV-1 Vpr: identification of determinants essential for subcellular localization.

Vpr is a conserved HIV-1 auxiliary protein that localizes to the nuclear region of cells. Vpr is also present in virions, and it is directed into the assembling virus when coexpressed with Gag. Each of these two localization activities may be important for Vpr function, and we recently identified regions of Vpr that are critical for virion incorporation. In this study we analyzed the Vpr domains involved in subcellular localization. Immunofluorescence staining of transfected cells showed that wild-type Vpr localized exclusively to the nuclear region. Mutations in the N-terminal domain that were designed to disrupt a predicted alpha-helical structure resulted in aberrant localization, while conservative substitutions showed a wild-type pattern. A region in the central portion of the protein also has the potential for helical structure, and mutagenesis of two conserved amino acids in this domain (A59, H71) impaired localization, while substitution of a third (Q65) did not. In contrast, neither the conserved Gly and Cys at positions 75-76 nor the C-terminal basic residues (R87, K95) were necessary for nuclear localization. In addition, two-residue insertions within and between the two putative helices disrupted localization but insertion in the C-terminal region did not. Thus, Vpr's subcellular localization function depends on the two putative helical domains but is independent of the conserved Gly-Cys motif and of specific C-terminal basic residues.

Amino Acid Sequence↗
PubMed1995

Role of the conserved dipeptide Gly75 and Cys76 on HIV-1 Vpr function.

Vpr is one of the accessory proteins encoded by the HIV-1 genome. Several interesting features associated with Vpr include incorporation into virus particles, ability to oligomerize, localization in the nucleus, and positive effect on virus production and replication. In order to understand the structure-function relationship of Vpr, we have analyzed the role of the Gly75 and Cys76 (GC) residues which are highly conserved in HIV-1 Vpr and in Vpr and Vpx of HIV-2/SIV. We have generated several substitution mutants involving this dipeptide and have evaluated for expression, stability, nuclear localization, and virion incorporation of Vpr. Our data demonstrate that the GC residues are not essential for virion incorporation and nuclear localization of Vpr. Serine substitution for Cys, however, restricted the localization of Vpr in the cytoplasm without affecting the Gag-directed incorporation of Vpr into virus-like particles. Interestingly, the cysteine-substituted mutants showed altered stability in comparison to the wild type, and substitution mutants for glycine showed minimal effect on stability. These results indicate that the glycine and cysteine do not play a role in nuclear localization or virion incorporation properties of Vpr and further suggest that these two functions of Vpr may not be interdependent.

Amino Acid Sequence↗
PubMed1995

Sequence of the dog immunoglobulin alpha and epsilon constant region genes.

The immunoglobulin alpha (IGHAC) and epsilon (IGHEC) germline constant region genes were isolated from a dog liver genomic DNA library. Sequence analysis indicates that the dog IGHEC gene is encoded by four exons spread out over 1.7 kilobases (kb). The IGHAC sequence encompasses 1.5 kb and includes all three constant region coding exons. The complete exon/intron sequence of these genes is described.

Amino Acid Sequence↗
PubMed1995

A miniaturized rapid paper chromatographic procedure for quality control of technetium-99m sestamibi.

A miniaturized rapid paper chromatographic technique (MRPC) for quality control of a technetium-99m sestamibi preparation was developed and compared with the manufacturer's and Hung et al. techniques. The MRPC system involves the use of 6.0x0.5cm Whatman 3MM paper strip developed in ethyl acetate. The procedure was completed within 3min while that of the manufacturer and Hung techniques took 30-35 and 4min respectively. The Rf range of 99mTc-sestamibi using MRPC was 0.55-0.75 while that of the other two techniques was 0.9-1.0. The results indicate that MRPC can be used to separate 99mTc-sestamibi from any 99mTc contaminant that migrates with the solvent front. The MRPC is a fast and effective chromatographic technique for routine quality control testing of 99mTc-sestamibi preparation.

Chromatography, Paper↗
PubMed1995

Decreased levels of salivary prostaglandin E2 and epidermal growth factor in recurrent aphthous stomatitis.

Prostaglandin E2 and epidermal growth factor are two important cytoprotective compounds in saliva. This study investigated their salivary levels in controls and individuals with minor recurrent aphthous stomatitis. The development of recurrent aphthous stomatitis was divided into three stages: (1) early active stage (mucosal redness); (2) active stage (mucosal ulceration); (3) convalescent stage. Unstimulated mixed saliva was collected from each volunteer. Salivary prostaglandin E2 and epidermal growth factor concentrations were determined by radioimmunoassay. Their levels (mean +/- SEM) were significantly lower during the active stage of ulceration as compared to the control: (a) for prostaglandin E2, 200 +/- 55 versus 73 +/- 11 pg/mg salivary protein (p < 0.01), 447 +/- 123 versus 112 +/- 19 pg/ml saliva (p < 0.01), 215 +/- 30 versus 63 +/- 12 pg/min salivary flow (p < 0.01), control (n = 12) versus active stage (n = 15); (b) for epidermal growth factor, 1.09 +/- 0.17 versus 0.67 +/- 0.17 ng/mg salivary protein (p < 0.05); 2.51 +/- 0.53 versus 0.84 +/- 0.19 pg/ml saliva (p < 0.05), 1.24 +/- 0.26 versus 0.41 +/- 0.09 pg/min salivary flow (p < 0.05), control (n = 12) versus active stage (n = 12). Salivary prostaglandin E2 and epidermal growth factor showed stage-dependent alterations during the development of the stomatitis. The prostaglandin E2 concentration decreased significantly during the active stage of ulceration, and then increased significantly during the convalescent stage. However, the recovery of salivary epidermal growth factor after the ulceration was slower than that of the prostaglandin E2. It is suggested that the diminution of prostaglandin E2 and epidermal growth factor in the saliva may be associated with the ulcer development.

Adult↗
PubMed1995

Modifications of the hydrophilicity of heterocyclic methacrylate copolymers for protein release.

A series of copolymers comprising ethyl methacrylate (EM) and tetrahydrofurfuryl methacrylate (THFMA) gelled with either THFMA monomer or hydroxyethyl methacrylate (HEMA) monomer have been developed. In this paper, we examine the water uptake characteristics of the polymer systems and address the possibility of increasing the hydrophilicity of the systems by changing the ratios of the copolymers. We have investigated whether protein release from the polymers is related to the composition of the polymer systems. More protein was released from the polymers gelled with the more hydrophilic monomer (HEMA) than with THFMA. This was consistent with the calculated diffusion coefficients, which were 10 times greater for the polymers gelled with HEMA than those gelled with THFMA. Interestingly, the water uptake and protein release profiles were not dependent on the ratio of EM and THFMA in the copolymers. This is probably due to the conflicting roles of THFMA in the copolymer; it is both the more hydrophilic component as well as a cross-linking agent. In addition, it would appear that the structural and surface topography of these polymers had more significant effects on protein release than copolymer composition.

Absorption↗
PubMed1995

Cellular colocalization of diuretic peptides in locusts: a potent control mechanism.

Locust abdominal ganglia are shown to colocalize Locusta-diuretic peptide-, leucokinin I-, and lysine vasopressin-like immunoreactivity in posterior lateral neurosecretory cells. Extracts of abdominal ganglia were partially purified by RP-HPLC then dot immunoassay screened with the same antisera used for immunocytochemistry. Locusta-diuretic peptide-like immunoreactive material coeluted with synthetic Locusta-diuretic peptide, and leucokinin-like immunoreactive material coeluted with locustakinin. Lysine vasopressin-like material eluted in fractions that also showed Locusta-diuretic peptide and leucokinin I immunoreactivity. The diuretic activity of synthetic Locusta-diuretic peptide and locustakinin is demonstrated, and they are shown to act at least additively to promote Malpighian tubule fluid secretion. The immunoreactive neurosecretory cells are assumed to express at least these two peptides, and a model for promoting fluid secretion is proposed.

Abdomen↗
PubMed1995

Biotinylated aprotinin: a versatile probe for the detection of serine proteinases on western blots.

The present study was undertaken to provide a highly sensitive detection system for the identification and characterisation of serine proteinases separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Biotinylated aprotinin of high specific activity (88-92% active) was prepared (i) by reaction of aprotinin directly with N-hydroxy sulfosuccinimidyl-6-(biotinamido) hexanoate, and (ii) by reaction of aprotinin-trypsin complex with N-hydroxy succinimidobiotin. Both biotinylated aprotinin samples were suitable as probes for the detection of the serine proteinases, neutrophil elastase and cathepsin G, pancreatic trypsin and chymotrypsin and plasmin on nitrocellulose blots. Specific irreversible chloromethyl ketone proteinase inhibitors used in combination with this detection system enabled respective proteinases to be selectively inactivated and thus positively identified. The biotinylated aprotinin detection system was highly sensitive and could detect as little as 0.2 ng (8.5 fmol) of active proteinase (trypsin). In summary, a method has been developed for the sensitive detection of serine proteinases separated by SDS-PAGE. The method is more sensitive and convenient to perform than conventional zymography and significantly, when used in conjunction with specific serine proteinase inhibitors or specific antibodies can yield appreciable information on the identity of the respective serine proteinases being examined. Furthermore the molecular mass of the serine proteinase may be reliably obtained by this method. This method should find application in identifying the role that serine proteinases play in the etiopathogenesis of connective tissue disorders.

Amino Acid Sequence↗
PubMed1995

Health insurance status and the use of emergency and other outpatient services by adults with sickle cell disease.

STUDY OBJECTIVE: To evaluate insurance status and frequency of use of emergency services in adults with sickle cell disease. DESIGN: Retrospective analysis of visits. SETTING: Emergency department and outpatient clinics of an urban university hospital. PARTICIPANTS: One hundred seventy-two subjects, who made 771 visits to the ED during 1990. RESULTS: Of the 172 subjects, 31 were covered by commercial insurance, 32 were covered by Medicare, and 109 were covered by Medicaid or were uninsured. Insurance status and frequency of use of emergency services were independent (P > .05). On discriminant analysis, Medicaid-covered and uninsured subjects were correctly classified, but commercially insured and Medicare subjects were not. Medicaid and uninsured subjects were more likely to be younger and to live closer to the hospital (P < .00005). High-frequency users of emergency services were discriminated from low-frequency users. High-frequency users were more likely to be younger, to be users of primary-care services, and to live closer to the hospital (P = .0004). CONCLUSION: Provision of primary-care services or stable insurance in the form of commercial insurance or Medicare did not decrease use of emergency services in subjects with sickle cell disease in a group of patients selected from one urban academic ED.

Adult↗
PubMed1995

Traumatic aneurysm of the maxillary artery: the role of interventional radiology. A report of two cases.

Two cases of post-traumatic aneurysm of the maxillary artery are described. The first patient was a 20-year-old man who sustained a Le Fort III type fracture in a road traffic accident. He experienced two episodes of significant maxillofacial haemorrhage, the first following admission and the second 5 days after initial reduction and fixation of his midfacial fractures. The second patient was a 23-year-old man with a bilateral cleft palate and extreme midfacial hypoplasia who underwent Le Fort I osteotomy. Significant bleeding commenced 3 h postoperatively and was not completely controlled by anterior and posterior nasal packing. Both the aneurysms were diagnosed on selective carotid angiography and successfully treated by embolization.

Adult↗
PubMed1995

Seizures and 'disappearing' brain lesions.

BACKGROUND--Patients with seizures may have abnormal brain imaging. Lesions demonstrated on computerized tomography or magnetic resonance imaging often suggest the underlying cause of the seizures, and may prompt an invasive investigation for diagnosis and treatment. In an increasingly recognized subset of patients however, the imaged lesions spontaneously resolve. While poorly understood, these 'disappearing' lesions may in fact be the consequence of seizures, rather than the cause. Two patients with 'disappearing' lesions are presented: the first patient had a proven cause, the second a probable one. Case 1. A patient with new onset seizures had an abnormal MRI. An angiogram confirmed thrombosis of the straight sinus. On follow-up, she was doing well; repeat imaging was normal with resolution of the lesion. Case 2. A patient with a long seizure history had been well controlled on medications, and had no change in frequency of her seizures. She was found to have a left homonymous hemianopsia. MRI revealed a right parieto-occipital lesion. The patient refused further work-up and was lost to follow-up. She returned almost a year later; repeat imaging was normal. CONCLUSION--Patients with seizures and abnormal imaging suggestive of structural lesions may occasionally have lesions that resolve spontaneously. While some diagnoses remain elusive, an important consideration to exclude in these patients is venous thrombosis.

Adult↗
PubMed1995

Invasive colitis and hepatitis due to previously uncharacterized spirochetes in patients with advanced human immunodeficiency virus infection.

We describe three patients with advanced human immunodeficiency virus (HIV) infection, two with colitis and one with cholestatic hepatitis, for whom results of detailed endoscopic and histologic studies were suggestive of invasive spirochetosis. In the two patients with colitis, colonoscopic evaluation revealed either diffuse ulcerations or pustules; in both cases, there was histologic evidence of extensive superficial cell necrosis and infiltration of the mucosa and lamina propria with acute inflammatory cells. Spirochetes in the mucosa and crypts were visualized by Warthin-Starry silver staining. Morphologically similar spirochetes throughout the liver specimen from the patient with cholestatic hepatitis were demonstrated by Warthin-Starry silver staining. Analysis with electron microscopy revealed these organisms to be loosely coiled spirochetes. Despite extensive evaluation, no other pathogens were identified. Invasive spirochetal infection, as defined by the results of Warthin-Starry silver staining of involved tissues, should be considered in the differential diagnosis of patients with HIV infection who have otherwise unexplained colitis or cholestatic hepatitis.

AIDS-Related Opportunistic Infections↗