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Biomedical subjects

M Saini

Publications and source records attributed to M Saini.

At least 37 records · Page 2Linked to original sources

Cloning and expression of bovine herpesvirus-1 glycoprotein C.

Glycoprotein C (gC) of Bovine Herpesvirus-1 (BHV-1) is expressed at high levels on surface of infected cells and on virus envelope. It is relatively immunodominant in antibody response to BHV-1 infection and protective in immunized bovines against BHV-1 challenge. In an attempt to express gC in mammalian cells, the 2.4 kb BamHI-EcoRI fragment, containing complete coding sequence of the gC gene was excised from a recombinant plasmid and cloned under the control of RSV-LTR. The resultant plasmid pRSV-gC was transfected into MDBK cells and expression of gC was detected by indirect immunofluorescence. The non-permeabilized cells revealed surface expression of gC.

Animals↗

Molecular characterization of an Indian isolate of Japanese encephalitis virus that shows an extended lag phase during growth.

The biological properties of an Indian isolate (GP78) of Japanese encephalitis virus (JEV) were characterized in tissue-cultured cells and mice and these were compared with the JaOArS982 strain from Japan. The GP78 strain had a markedly extended lag phase during its growth in porcine stable kidney (PS) cells. There were no obvious defects in the penetration of GP78 into PS cells. However, viral RNA and protein synthesis were significantly delayed in GP78-infected PS cells. Fusion-from-within assays carried out in C6/36 cells indicated that GP78 was less fusogenic than the JaOArS982 strain of JEV. Moreover, maximum fusion in GP78-infected cells occurred at pH 5.5, whereas JaOArS982-infected cells showed maximum fusion at pH 6.0. These results suggested that there may be a lesion in the virus-cell fusion process. The GP78 strain also showed delayed growth in brains of 1-week-old BALB/c mice. Although JEV GP78 was as virulent as the JaOArS982 strain in these mice, the appearance of clinical symptoms of JEV infection was delayed by a day in mice infected with the GP78 strain and these animals showed an increased average survival time. Comparison of the nucleotide sequences of the GP78 and the JaOArS982 strains of JEV identified a number of amino acid substitutions in structural proteins. Of these, a Thr --> Met substitution at residue 76 of the envelope protein is predicted to be causally associated with the altered biology of the GP78 strain during growth.

Amino Acid Sequence↗

Induction of nitric oxide in bovine peripheral blood mononuclear cells by bovine herpesvirus 1.

Bovine peripheral blood mononuclear cells (PBMCs) and monocytes were stimulated with bovine herpesvirus 1 (BHV-1), LPS and concanavalin A (Con A) to produce L-arginine-dependent nitric oxide (NO) in vitro. NO was detected as early as 12 hrs and up to 72 hrs post stimulation (p.s.). The NO from lipopolysacharide (LPS)-stimulated PBMCs and monocytes was found to exhibit antiviral effect against BHV-1. The anti-BHV-1 effect was inhibited with N(omega)-methyl L-arginine indicating involvement of inducible NO synthase (iNOS) in NO production.

Animals↗

Apoptosis in bovine herpesvirus-1 infected bovine peripheral blood mononuclear cells.

Apoptosis is a process whereby cells die in a controlled manner in response to various stimuli like cytotoxins, viral antigens and normal physiological signals during differentiation and development. Virus induced immunosuppression has been reported for various viral diseases including Bovine Herpesvirus-1 (BHV-1). In the present study, BHV-1 was found to cause apoptosis in ConA stimulated bovine peripheral blood mononuclear cells (PBMCs). Apoptotic index quantified by fluorescent dyes revealed a significant (P < 0.001) increase in percent apoptotic cells at 2, 24 and 48 hr post infection as compared to their respective non-infected controls. Apoptosis specific internucleosomal laddering in DNA from BHV-1 infected PBMCs was seen in agarose gel electrophoresis. No DNA fragmentation was observed in control non-infected PBMCs.

Animals↗

Use of prokaryotically expressed nucleocapsid protein as positive antigen in ELISA.

A cDNA library of Rinderpest vaccine virus was prepared in Zap Express vector (Stratagene). The Rinderpest 'N' gene specific clones were selected, characterized and thereafter expressed in E. coli XLOLR strain. The expressed protein was found to be immunogenic in western blot with hyperimmune sera. It reacted with rinderpest and 'N' protein specific monoclonal antibodies in Enzyme Linked Immunosorbent Assay (ELISA). Prokaryotically expressed 'N' protein also gave precipitin band in counter immunoelectrophoresis test (CIE). The expression of N protein was sufficient for its utility as positive antigen in CIE and ELISA used for rinderpest diagnosis.

Animals↗

The vertical displacement of the center of mass during walking: a comparison of four measurement methods.

Measuring the vertical displacement of the center of mass (COM) of the body during walking may provide useful information about the energy required to walk. Four methods of varying complexity to estimate the vertical displacement of the COM were compared in 25 able-bodied, female subjects. The first method, the sacral marker method, utilized an external marker on the sacrum as representative of the COM of the body. The second method, the reconstructed pelvis method, which also utilized a marker over the sacrum, theoretically controlled for pelvic tilt motion. The third method, the segmental analysis method, involved measuring motion of the trunk and limb segments. The fourth method, the forceplate method, involved estimating the COM displacement from ground reaction force measurements. A two-tailed paired t-test within an ANOVA showed no statistically significant difference between the sacral marker and the reconstructed pelvis methods (p = 0.839). There was also no statistically significant difference between the sacral marker and the segmental analysis method (p = 0.119) or between the reconstructed pelvis and the segmental analysis method (p = 0.174). It follows that the first method, which is the most simple, can provide essentially the same estimate of the vertical displacement of the COM as the more complicated second and third measures. The forceplate method produced data with a lower range and a different distribution than the other three methods. There was a statistically significant difference between the forceplate method and the other methods (p < 0.01 for each of the three comparisons). The forceplate method provides information that is statistically significantly different from the results of the kinematic methods. The magnitude of the difference is large enough to be physiologically significant and further studies to define the sources of the differences and the relative validity of the two approaches are warranted.

Adult↗

Bovine herpesvirus type 1 glycoprotein C expression in MDBK cells and its reactivity in enzyme-linked immunosorbent assay.

Glycoprotein C (gC) of bovine herpesvirus type 1 (BHV-1) is a major viral glycoprotein expressed at high level on the surface of infected cells and on the virion envelope. This glycoprotein is also a major target of immune response at both humoral and cellular levels. The plasmid pRSV-gC having complete coding gene for BHV-1 gC was transfected into MDBK cells and the expression of gC in these cells was detected by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) and immunoblot analysis. Transcription of the gC gene was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) of total RNA isolated from transfected cells. MDBK cells expressing BHV-1 gC were used as antigen in enzyme-linked immunosorbent assay (ELISA) of antibodies against BHV-1 in field sera. The results were found comparable (92.44%) with those obtained with BHV-1 purified antigen.

Animals↗

Human serum immunoglobulin counteracts rotaviral infection in Caco-2 cells.

Oral administration of human serum immunoglobulin reduces the duration of diarrhea and of rotaviral excretion in children. To investigate the in vitro effects of immunoglobulin on virus-enterocyte interaction, Caco-2 cells were infected with Rotavirus strain SA11. Immunoglobulin was added prior to and at various times postinfection. Indirect immunofluorescence was performed with an antibody against VP-6 rotaviral antigen. Cell viability and monolayer transepithelial electrical resistance (TEER) were monitored. Immunofluorescence showed a perinuclear distribution in 90% of cells. Rotavirus infection induced a progressive decrease in TEER and a parallel reduction in cell viability, depending on viral load. Preincubation of the virus with immunoglobulin prevented cell infection as judged by immunofluorescence. Immunoglobulin addition to infected cells partially prevented the decrease in TEER and induced a later shift of TEER toward increasing values, suggesting restoration of monolayer's integrity. The efficacy of immunoglobulin depended on its concentration and on the time of its addition. These results indicate that immunoglobulin is effective in preventing infection and in reducing cell damage, through a direct anti-Rotavirus action and may indicate that immunoglobulin should be administered in the early phase of diarrhea, to reduce the severity of Rotavirus infection.

Administration, Oral↗

Hematological alterations in propylene glycol-dosed female rats are minimal.

The effect of a single oral dose of 73 or 294 mg propylene glycol/100 g bw, in Group I or II respectively, on hematological parameters, plasma osmolality and erythrocyte morphology was investigated in groups of 6 female rats each at different time intervals after dosing. A statistically significant and progressive decrease was observed in hemoglobin, packed cell volume and red cell counts for 2 d, which returned to basal values on the 8th day. Reticulocyte counts, plasma hemoglobin and osmolality increased after PG dosing in both the groups, and the changes were more pronounced after 2 d. The osmotic fragility of erythrocytes remained unaffected after PG dosing whereas electron microscope morphology revealed rough cell surface, ruptured membranes and increased cell adherence throughout the observation period, but these features were not marked on the 8th day. These modified red cell surface characteristics could promote removal by the reticulo-endothelial system resulting in the significant increases in spleen weight in both groups.

Administration, Oral↗

Isolation of a spermatozoa motility inhibiting factor from chicken seminal plasma with antibacterial property.

A 78-kDa spermatozoa motility inhibiting factor (SMIF) was purified from chicken (Gallus domesticus) seminal plasma by anion exchange (DE-53) followed by affinity chromatography on concanavalin A-Sepharose. The factor is thermostable and inhibited the spermatozoa motility in a dose dependent manner. In addition, SMIF inhibited the growth of gram negative bacteria, Pasteurella multocida but not gram positive Streptococcus equi. The factor lost its spermatozoa immobilizing property after treatment with trypsin, chymotrypsin or pepsin. The inhibition of SMIF by beta-mercaptoethanol suggest the involvement of disulfide bonds in its activity. Similarly, this property was lost in presence of chicken seminal plasma or incubating SMIF with anti-SMIF antibodies. Evidence is provided for the presence of a high molecular weight protein (> 100 kDa) in chicken seminal plasma that neutralizes the motility inhibiting property of SMIF. No significant decrease in spermatozoa ATP was observed in presence of SMIF suggesting that the loss of spermatozoa motility was due to factors other than depletion in cell's energy. Using anti-SMIF antibodies, a cross-reactive protein was identified in the blood, liver and reproductive tissues of chicken and the seminal plasma of cattle and buffalo. However, the cross-reactive protein failed to inhibit chicken spermatozoa motility. The significance of SMIF in chicken seminal plasma is discussed.

Animals↗

Opposite polarity of virus budding and of viral envelope glycoprotein distribution in epithelial cells derived from different tissues.

We compared the surface envelope glycoprotein distribution and the budding polarity of four RNA viruses in Fischer rat thyroid (FRT) cells and in CaCo-2 cells derived from a human colon carcinoma. Whereas both FRT and CaCo-2 cells sort similarly influenza hemagglutinin and vesicular stomatitis virus (VSV) G protein, respectively, to apical and basolateral membrane domains, they differ in their handling of two togaviruses, Sindbis and Semliki Forest virus (SFV). By conventional EM Sindbis virus and SFV were shown to bud apically in FRT cells and basolaterally in CaCo-2 cells. Consistent with this finding, the distribution of the p62/E2 envelope glycoprotein of SFV, assayed by immunoelectronmicroscopy and by domain-selective surface biotinylation was predominantly apical on FRT cells and basolateral on CaCo-2 cells. We conclude that a given virus and its envelope glycoprotein can be delivered to opposite membrane domains in epithelial cells derived from different tissues. The tissue specificity in the polarity of virus budding and viral envelope glycoprotein distribution indicate that the sorting machinery varies considerably between different epithelial cell types.

Animals↗

The relationship of various indices of heart size on chest x-ray to the 10-year incidence of hypertension. The Normative Aging Study.

A total of 598 males (aged 30-74 years) who had baseline (1961-1970) chest radiography and baseline blood pressure less than 140/90 mmHg were observed prospectively for 10 years. Subjects were participants of the Normative Aging Study, a longitudinal study on aging initiated in 1961 at the Veterans Administration Outpatient Clinic in Boston, Massachusetts. Blood pressures were taken at five- and 10-year follow-up examinations. Multiple logistic regression analysis indicated that the long diameter of the heart (on posteroanterior film) and the cardiac depth (on lateral film) were statistically significant predictors of subsequent hypertension after controlling for baseline body mass index, systolic pressure, and diastolic pressure. A similar model considering various composite indices of heart size indicated that the heart volume was a statistically significant and independent predictor of hypertension. Thus, increases in heart size may precede and predict the development of sustained hypertension.

Adult↗

Traumatic olecranon bursitis. Radiologic observations.

The affected elbow of 28 patients with traumatic olecranon bursitis was radiographically compared with the homologous elbow of 28 matched controls. Olecranon spurs, amorphous calcium deposits, or both, were present in 16 patients and 4 controls (p less than 0.01). Air was injected in the bursa in 12 additional patients. Nodules in the bursal floor were noted in 10, and the bursa was partially septated in 8. Olecranon spurs, present in 6 patients, corresponded to the insertion of the triceps tendon. With elbow flexion the bursa flattened and lengthened while the olecranon process glided distally beneath the bursal floor.

Adult↗

Revised radiologic concepts of the Barrett esophagus.

The radiologic findings in 39 cases of histologically proven Barrett esophagus are reviewed. Esophageal strictures were not found in 18% of the cases, and when present were more often distally located. Esophageal ulcers were found in only 46% of the patients. In 13% of the cases neither ulcer nor stricture was found. The majority (72%) of these cases did not fit the classic stereotype of high esophageal stricture and/or ulcer.

Epithelium↗