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Biomedical subjects

M Sharma

Publications and source records attributed to M Sharma.

At least 19 recordsLinked to original sources

Identification of a family of bacteriophage T4 genes encoding proteins similar to those present in group I introns of fungi and phage.

The bacteriophage T4 segA gene lies in a genetically unmapped region between the gene beta gt (beta-glucosyltransferase) and uvsX (recombination protein) and encodes a protein of 221 amino acids. We have found that the first 100 amino acids of the SegA protein are highly similar to the N termini of four other predicted T4 proteins, also of unknown function. Together these five proteins, SegA-E (similar to endonucleases of group I introns), contain regions of similarity to the endonuclease I-Tev I, which is encoded by the mobile group I intron of the T4 td gene, and to putative endonucleases of group I introns present in the mitochondria of Neurospora crassa, Podospora anserina, and Saccharomyces douglasii. Intron-encoded endonucleases are required for the movement (homing) of the intron DNA into an intronless gene, cutting at or near the site of intron insertion. Our in vitro assays indicate that SegA, like I-Tev I, is a Mg(2+)-dependent DNA endonuclease that has preferred sites for cutting. Unlike the I-Tev I gene, however, there is no evidence that segA (or the other seg genes) resides within introns. Thus, it is possible that segA encodes an endonuclease that is involved in the movement of the endonuclease-encoding DNA rather than in the homing of an intron.

Amino Acid Sequence

Cyst fluid from human autosomal dominant polycystic kidneys promotes cyst formation and expansion by renal epithelial cells in vitro.

Autosomal dominant polycystic kidney disease (ADPKD) is characterized by progressive renal enlargement, culminating in renal insufficiency in over one half of affected individuals. The highly variable onset and clinical course of ADPKD may be due to factors extrinsic to the genetically defined renal cysts. In this study, cyst fluid samples from 12 nonazotemic and 18 azotemic ADPKD subjects were examined for in vitro biologic activity that promotes cellular proliferation and the secretion of fluid by renal epithelial monolayers, two pathogenetic mechanisms that have critical roles in the formation and the rate of expansion of renal cysts. Cyst fluid added to culture medium (final concentrations, 1 to 20%) caused Madin-Darby canine kidney cells and human kidney cortex (HKC) cells derived from primary cultures to form cysts in Type I collagen matrix. Cyst fluid stimulated the net transepithelial secretion of fluid by polarized monolayers composed of these same cells. Absolute levels of fluid secretory activity determined by MDCK bioassay were correlated directly with the rate of fluid secretion by HKC cell monolayers and with the extent of cyst formation by MDCK and HKC cells embedded in collagen matrix. The secretory activity of urine was negligible; secretory activity was detectable in the serum of normal and ADPKD subjects, but the levels were much lower than in cyst fluid. cAMP agonists prostaglandins E1 and E2, arginine vasopressin, and 8-Br-cAMP stimulated fluid secretion by MDCK and HKC monolayers, but these substances did not cause HKC cells to form cysts in collagen matrix, whereas cyst fluid did. Among other naturally occurring growth factors and autacoids, only epidermal growth factor and transforming growth factor alpha stimulated cyst formation by HKC cells; however, the capacity of cyst fluid to stimulate fluid secretion was not affected by treatment with antiserum to epidermal growth factor. It was concluded that potent, and possibly unique, substances in the cyst fluids of individuals with ADPKD support and augment biologic processes in renal epithelial cells that may be important in the promotion of progressive cyst expansion.

8-Bromo Cyclic Adenosine Monophosphate

Methotrexate: clinical and immunological effects in refractory rheumatoid arthritis.

Thirty five patients with refractory rheumatoid arthritis were given 7.5 mg of methotrexate (Mtx) every week. Eleven patients had to discontinue treatment either because of adverse effects or unresponsiveness. Twenty four patients showed clinical response and significant improvement in ESR and they continued Mtx for a mean of 25.24 months. Seven patients achieved clinical remission as defined by ARA criteria. Immunological parameeters including IgG, IgM, IgA, lymphocyte subsets (CD3+, CD4+, CD8+ and B), C3 and C4 however, did not show any change during this treatment in any of the groups upto 6 months. There was a significant fall in the erythrocyte sedimentation rate (ESR), c-reactive protein (CRP) and rheumatoid factor (RF) levels in responders only.

Administration, Oral

Gamma glutamyl transpeptidase in meningitis.

Gamma glutamyl transpeptidase (GGTP) was measured serially in cerebrospinal fluid (CSF) and serum in 23 cases of meningitis (15 pyogenic and 8 tuberculous meningitis) and an equal number of age and sex matched healthy controls, to find out its diagnostic and prognostic significance in meningitis. GGTP activity was significantly elevated (p less than 0.001) in CSF and serum in meningitis as compared to control subjects. Levels were significantly higher in pyogenic as compared to tuberculous meningitis (p less than 0.001) and in CSF than in serum (p less than 0.001). The maximum elevation was seen on the 1st day and thereafter the activity declined in the majority (65.2%) of cases. However, in 3 cases of pyogenic meningitis and 5 cases of tuberculous meningitis, the GGTP activity on subsequent estimation increased serially; all these 8 cases died. It is concluded that CSF GGTP activity is significantly elevated in meningitis and serial rise in its activity is associated with poor prognosis and even fatal outcome.

Adult

Wallagotrema indicus n.sp., a new monogenean from the freshwater sheat-fish Wallagu attu Bloch and Schneider.

The paper describes Wallagotrema indicus n.sp., a freshwater monogenean recovered from the gill filaments of the freshwater sheat-fish Wallago attu Bloch and Schn. It is characterized by the number of head organs, an accessory piece on the cirrus, wings on the anchors and a beak-like outgrowth on the dorsal anchors, together with numerous other differences in the shape and size of various body parts. On the basis of these findings, the generic diagnosis of Wallagotrema is amended and a key to the various species of this genus is appended.

Animals

Role of Chlamydia trachomatis in pelvic inflammatory disease.

Endocervical swabs and peritoneal aspirate from the pouch of Douglas (POD) were taken in 50 women with pelvic inflammatory disease (PID) and in 50 controls for the detection of C. trachomatis antigen (Chlamydiazyme). C. trachomatis antigen was detected from endocervix in 8 per cent patients and 2 per cent controls (P greater than 0.05), while from the POD it was detected in 44 per cent patients and in none of the control women (P less than 0.005). There was a higher detection rate of C. trachomatis antigen from the POD in women with chronic PID (50%) as compared to acute PID (28.5%; P greater than 0.05). Culdocentesis was found to be a simple and reliable OPD procedure for obtaining material for C. trachomatis antigen detection.

Adult

Aetiologic factors in male infertility: clinical, microbiological and hormonal evaluation.

Seventy two infertile men were studied. History of small pox and mumps infection was noted in 4 and 3 patients respectively. Seven patients had varicocele (9.2%), and small atrophic testes were found in 9 (12.5%). Azoospermia was reported in 41 (58.3%) and oligospermia in 17 (23.6%), and 14 patients (19.4%) had normal sperm counts. Mycoplasma were grown from urethral swabs in 25 (35%) patients. Mean LH and FSH were elevated in azoospermics (p less than 0.001), E2-17B in oligospermics (p less than 0.001) and FSH in normospermic (p less than 0.01) patients. Hypergonadotropism suggestive of primary testicular failure was recorded in 43 (59.7%) patients. Hypogonadotropism was noted in 3 (4%) and hyperprolactinemia due to pituitary microadenoma induced infertility in only one patient. No aetiology could be determined in 11 (16%) patients.

Adult

Nuclease P1-mediated fluorescence postlabeling assay of AAF modified DNA model d(TACGTA) and calf-thymus DNA.

Fluorescence postlabeling assay for DNA damage combines enzymatic digestion of modified DNA to nucleoside monophosphates and fluorescence postlabeling. However, to facilitate the quantitative release of bulky adducts that are not readily obtained as mononucleotides, a different mechanism is essential. In order to test the application potential of nuclease P1-mediated fluorescence postlabeling to assay bulky adduct, d(TACGTA) was reacted with N-acetoxy-2-acetylamino-fluorene. The major product, characterized by nmr as AAF adduct of the guanine moiety at the C-8 position, was used as a DNA model. Nuclease P1 digestion of the modified oligomer excised the adduct in two forms (3:1) which were identified as AAF modified d(pGpT) and dpG respectively by cochromotography with authentic markers. Fluorescence postlabeling assay of AAF modified d(TACGTA) detected both forms of the excised adduct. The application of the overall procedure to assay AAF modified calf-thymus DNA demonstrated that the extension of fluorescence postlabeling technique from the mononucleotide to the dinucleotide version expands the scope of the assay.

Acetoxyacetylaminofluorene

Micellar electrokinetic capillary chromatographic separation and laser-induced fluorescence detection of 2'-deoxynucleoside 5'-monophosphates of normal and modified bases.

Micellar electrokinetic chromatography is used to separate dansylated nucleotides, both normal and modified species. The high separation power allows detection of minor components present in less than 1 part per thousand of the major components. Laser-excited fluorescence is used to detect the separated components at the 6 x 10(-18) mol level or 10(-9) M injected material. Combined with high-performance liquid chromatographic enrichment prior to labeling, this technique can be used to assess DNA damage in carcinogenesis studies.

Animals

Effect of chronic microwave radiation on T cell-mediated immunity in the rabbit.

Experiments were conducted to elucidate the effects of chronic low power-level microwave radiation on the immunological systems of rabbits. Fourteen male Belgian white rabbits were exposed to microwave radiation at 5 mW/cm2, 2.1 GHz, 3 h daily, 6 days/week for 3 months in two batches of 7 each in specially designed miniature anechoic chambers. Seven rabbits were subjected to sham exposure for identical duration. The microwave energy was provided through S band standard gain horns connected to a 4K3SJ2 Klystron power amplifier. The first batch of animals were assessed for T lymphocyte-mediated cellular immune response mechanisms and the second batch of animals for B lymphocyte-mediated humoral immune response mechanisms. The peripheral blood samples collected monthly during microwave/sham exposure and during follow-up (5/14 days after termination of exposures, in the second batch animals only) were analysed for T lymphocyte numbers and their mitogen responsiveness to ConA and PHA. Significant suppression of T lymphocyte numbers was noted in the microwave group at 2 months (P less than 0.01, delta % 21.5%) and during follow-up (P less than 0.01, delta % 30.2%). The first batch animals were initially sensitised with BCG and challenged with tuberculin (0.03 ml) at the termination of microwave irradiation/sham exposure and the increase in foot pad thickness (delta mm), which is a measure of T cell-mediated immunity (delayed type hypersensitivity response, DTH) was noted in both the groups. The microwave group revealed a better response than the control group (delta % +12.4 vs. +7.54). The animals were sacrificed and the tissue T lymphocyte counts (spleen and lymph node) were analysed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Atrial septal defect presenting as recurrent primary amoebic lung abscess.

A middle-aged female with an atrial septal defect (secundum type) presented with a primary pulmonary amoebic abscess. She was successfully treated with antiamoebic therapy. One year later she presented with a similar lung abscess which again responded to antiamboebic treatment. Attention is drawn to the fact that a patient with a left to right shunt can present with a recurrent rare primary parasitic infection of the lung.

Amebicides

Seminal vesiculitis due to Mycobacterium gastri leading to male infertility.

Tuberculosis of the genitourinary tract is a well-known cause of male infertility. However, infertility from infection by nontuberculous mycobacterium has not been reported. Herein, we present a case of seminal vesiculitis due to Mycobacterium gastri in a diabetic patient leading to male infertility. Improvement in semen quality was noticed after 6 months of therapy with isoniazid, ethambutol and rifampicin.

Adult

Protective immunity to entamoeba histolytica infection in subjects with antiamoebic antibodies residing in a hyperendemic zone.

To investigate the relationship between amoebic gut infection and the presence of serum antiamoebic antibodies in a community where amoebiasis is highly endemic, a cross-sectional population survey was conducted covering 982 subjects in 2 adjacent villages in rural India. E. histolytica was detected in fecal smears in 18.1% (88/487) and antiamoebic antibodies by micro-ELISA in 28.7% (251/874) subjects. In 450 subjects in whom there was no clinical evidence of invasive amoebic disease or history of ingestion of antiamoebic drugs in the previous year and from whom both stool and matched serum samples were obtained, correlation of gut infection with seropositivity was attempted. Seropositivity in 80 cyst passers (23.8%) was significantly lower than in 370 non-cyst passers (35.1%; p less than 0.05). Conversely the rate of gut infection with E. histolytica was lower in seropositive subjects (19/149; 12.8%) as compared to seronegative subjects (61/301; 23%, p less than 0.05). In the population where E. histolytica infection was highly endemic, the rate of gut infection with the parasite was inversely related to seropositivity. Our data suggests that invasive E. histolytica infection (mostly asymptomatic) evokes good gut immunity in the host with clearing of the parasite from the colon and/or resistance to reinfection. High prevalence rate of amoebic antibodies indicates good "herd" immunity.

Adolescent