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Biomedical subjects

R Sharma

Publications and source records attributed to R Sharma.

At least 307 records · Page 17Linked to original sources
PubMed1996

Alteration of glomerular permeability to macromolecules induced by cross-linking of beta 1 integrin receptors.

Altered glomerular epithelial cell attachment to the glomerular basement membrane is an important pathogenetic factor in increased glomerular permeability to proteins. We have previously presented evidence that antibodies reactive with integrin matrix receptors on glomerular epithelial cells inhibit adhesion of these cells and may be involved in the production of proteinuria in vivo. Therefore, we utilized intact glomeruli in an in vitro system to directly assess the effect of anti-beta 1-integrin antibody on glomerular permeability. Permeability to albumin (Palb) was calculated from the volume response of glomeruli to a transcapillary oncotic gradient. Anti-beta 1-integrin increased Palb in a dose- and time-dependent manner. Palb was increased to 0.70 +/- 0.05 whereas normal rabbit IgG had no effect (0.10 +/- 0.04). F(ab')2 fragments of antibody increased Palb to a similar degree whereas Fab fragments had no effect (0.10 +/- 0.06). Cross-linking of Fab fragments, however, with a second antibody restored their ability to increase Palb (0.60 +/- 0.09), demonstrating the importance of integrin cross-linking in producing the observed effect. Intact, F(ab')2 and Fab fragments of anti-beta 1 antibody all inhibited adhesion of glomerular epithelial cells to fibronectin, laminin, and types I and IV collagen, although the degree of inhibition by Fab fragments was significantly less on collagens. No cytotoxic effects were observed with anti-beta 1 antibody or its fragments. These results suggest that antibodies to integrin matrix receptors on glomerular cells alter cell interactions with the glomerular basement membrane and lead to increased glomerular permeability to proteins via a process that is initiated by integrin cross-linking rather than through simple interference with cell adhesion per se.

Albumins↗
PubMed1996

Purification and kinetic characterization of chicken liver inorganic pyrophosphatase.

Soluble inorganic pyrophosphatase (EC 3.6.1.1) was isolated from chicken liver, RIR breed, to apparent homogeneity. The enzyme showed a molecular mass of 100 kDa as estimated by gel filtration and a subunit mass of 49 kDa on SDS-PAGE. The enzyme was very specific for pyrophosphate (PPi) and magnesium, and there was no measurable activity on replacing Mg2+ with Zn2+. At optimal conditions of assay, 50% of the enzyme activity was inhibited at 42 microM Ca2+, 70 microM fluoride and 0.91 mM Cd2+. There was a 50% inactivation of enzyme activity at 0.1 M guanidine hydrochloride (GuHCl). Kinetic analysis of GuHCl inactivation revealed 2 essential binding sites for this ligand. The enzyme showed allosteric behaviour with the substrate PPi and Mg2+. The apparent Hill coefficient of 1.47 and 1.48 for PPi and Mg2+, respectively indicate positive cooperatively. Hill plots also gave [S]0.5 of 0.177 mM and 2.5 microM for Mg2+ and PPi, respectively.

Animals↗
PubMed1995

Neuroprotective use-dependent blockers of Na+ and Ca2+ channels controlling presynaptic release of glutamate.

We have originated a family of N,N'-disubstituted guanidines that block the voltage-activated Ca2+ and Na+ channels governing glutamate release. These compounds, CNS 1237 (N-acenaphthyl-N'-methoxynaphthyl guanidine) and its analogues, are "use dependent" in their ability to attenaute neurotransmitter release: they block glutamate release with greater efficacy under conditions of persistent or repetitive depolarization, as would be encountered under pathophysiological circumstances, relative to their ability to block glutamate release elicited by brief, transient depolarizations more characteristic of normal physiological release events in nonischemic brain. Using electrophysiological and rapid kinetic methods, we have differentiated the use-dependent block of the relevant Na+ and Ca2+ channels governing neurotransmitter release from the mechanism of channel antagonism exhibited by, respectively, the substituted guanidine Na+ channel blocker tetrodotoxin (TTX) and venom peptide Ca2+ antagonists. To characterize use-dependent Na+ channel block by CNS 1237, we have employed whole-cell voltage-clamp recordings from a Chinese hamster ovary (CHO) cell line expressing cloned mammalian type II Na+ channels. These experiments demonstrated that, in contrast to the actions of TTX under the same conditions, the potency of Na+ channel block by CNS 1237 is greatly enhanced by depolarizing stimuli in a frequency-dependent manner. Ca2+ channel-activated glutamate release from brain nerve terminal preparations was measured with approximately 300 msec time resolution over a 5-second period of high K(+)-depolarization, using a rapid superfusion technique. CNS 1237 and analogues, at 1-3 microM, accelerated the decay of glutamate release by 40-70%, reflecting depolarization-induced enhancement of block. In contrast, blockade of glutamate release by the Ca2+ channel antagonist peptide toxins omega-aga IV-A (from spider venom) and omega-conotoxin M-VII-C (from cone snail venom) exhibited "reverse-use-dependence:" at concentrations of 0.3 microM, which blocked the initial amplitude of glutamate release by 40-60%, the decay time constant for glutamate release was significantly increased, indicating depolarization-induced relief of block. These findings establish that CNS 1237 and other members of this compound series are use-dependent blockers of the voltage-activated ion channels governing glutamate release. Studies of CNS 1237 in the rat middle cerebral artery occlusion (MCAO) focal stroke model have indicated infarct size reduction comparable to that observed by the same investigators for the glutamate release blocker (BW 619C89 (Burroughs-Wellcome, now in clinical development). Maximal infarct size reduction is achieved with a 3-mg/kg bolus followed by a 4-hour infusion of 0.75 mg/kg/hr.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1995

Structure-activity studies of RFamide analogues on central neurones of Helix aspersa.

The effects of FMRFamide were compared with those of FMRFamide analogues, FLRFamide, LFRFamide, FFRFamide, LLRFamide, D-FMRFamide, F-D-MRFamide and FM-D-RFamide, and the fragments, MRFamide and LRFamide, on identified central neurones, F1, F2, F5 and E16, of the snail Helix aspersa, using intracellular recording and two electrode voltage clamp techniques. All FMRFamide analogues showed an inhibitory effect on F1 neurones with an order of potency: FLRFamide > FMRFamide > FFRFamide > LFRFamide >> LLRFamide. FMRFamide and FLRFamide exhibited a biphasic response on F2 neurones. At lower concentrations (< 10 microM), both peptides usually only excited while at higher concentrations (> 30 microM), exhibited an excitation followed by an inhibition. FFRFamide only excited F2 while LFRFamide and LLRFamide only inhibited F2. LRFamide and MRFamide (100 microM) were inactive on both F1 and F2. FLRFamide, LFRFamide, LLRFamide, FFRFamide and D-FMRFamide showed cross-interaction on the outward current induced by FMRFamide in F5. All peptides induced an outward current and also reduced the FMRFamide-induced current reversibly. In contrast, MRFamide, LRFamide and F-D-MRFamide failed to have direct effects on these neurones nor interact with the FMRFamide-induced current. We conclude that on F2 neurones Phe is essential for the activation of the RFamide receptor mediating the excitation and Met or Leu are important to activate the RFamide receptors mediating the inhibition. Removal of the N-terminal Phe, to give LRFamide and MRFamide render the peptides inactive. Therefore a tetrapeptide sequence is essential for the biological activity of FMRFamide analogues on these Helix neurones. FLRFamide, LFRFamide, LLRFamide, FFRFamide and D-FMRFamide exhibit a cross-interaction with FMRFamide. It is possible that these peptides also act on the same class of RFamide receptors as agonists to cause cross desensitization.

Animals↗
PubMed1995

Developmental expression and corticosterone inhibition of adenosine deaminase activity in different tissues of mice.

The activity expression and corticosterone inhibition of adenosine deaminase (ADA) were studied in the spleen, stomach, and liver of mice at various postnatal ages. The specific activity of ADA is very low in the spleen and stomach of 5- and 10-day-old mice, and increases significantly (2.5- to 3.0-fold) in 20- and 30-day-old animals. Its level shows a further increase in the spleen of 60-day-old mice while stomach increase of ADA is not significant. In contrast, the activity of ADA is significantly higher in the liver of 5- and 10-day-old mice, decreases markedly (2.5-fold) in 20- and 30-day-old animals and shows a sharp increase in the liver of 60-day-old mice. Corticosterone administration brings a marked inhibition in the activity of ADA at all ages studied in the spleen and stomach whereas it inhibits the liver ADA only at 30 and 60 days postnatal age. These findings suggest an age- and tissue-specific expression of ADA activity and also indicate corticosterone as an inhibitory regulator of this enzyme.

Adenosine Deaminase↗
PubMed1995

Angiodysplasia and lower gastrointestinal tract bleeding in elderly patients.

Angiodysplasia of the colon is one of the most common causes of major lower intestinal tract bleeding in the elderly; it occurs predominantly in the cecum and on the right side of the colon and is thought to result from degenerative changes associated with aging. The clinical presentation is varied, ranging from hematochezia or melena to iron-deficiency anemia resulting from long-term blood loss. Accurate diagnosis may require a combination of diagnostic techniques, such as angiography, nuclear scanning, and colonoscopy. The management plan should be individualized for each patient depending on severity, rate of rebleeding, and issues of comorbidity. Although conservative medical management is a reasonable option for many patients, endoscopic treatment has generally replaced surgery as the first line of definitive treatment for angiodysplasias in most of these patients. The risk of rebleeding is a considerable problem, and surgical therapy yields better results in this aspect. The role of hormonal therapy is not clearly established.

Angiodysplasia↗
PubMed1995

Angiosarcoma of the breast. Initial misdiagnosis is still common.

Primary angiosarcoma of the breast is a rare and often misdiagnosed disease. The most common clinical presentation is a painless mass in the affected breast, but the often varied presentation and the high incidence of histologic misdiagnosis make early detection rare. The tumor size and the histologic type correlate with the prognosis. The treatment for angiosarcoma of the breast is early and complete surgical excision of the mass with adequate margins. Axillary dissection is not indicated because the predilection for nodal metastasis is rare. The definitive role of adjuvant therapy remains undetermined. Chemotherapy and radiotherapy may play an important role in survival; however, the data are inconclusive. A high index of suspicion for angiosarcoma is a crucial tool in its proper diagnosis and treatment. It should always be noted that a vascular lesion that is associated with any breast mass is an angiosarcoma until proven otherwise.

Adult↗
PubMed1995

Comparative evaluation of classical intrafascial-supracervical hysterectomy (CISH) with transuterine mucosal resection as performed by pelviscopy and laparotomy--our first 200 cases.

A novel way of performing endoscopic intrafascial supracervical hysterectomy is presented. By using the endoscopic approach for dissection as well as uterine extraction using the serrated-edged macro-morcellator, we avoid giving the patient a colpotomy incision and its inherent post-operative discomfort. A further modification involves nearly complete excision (95%) of uterocervical mucosa using a calibrated resection tool, thus eliminating the possible subsequent development of cervical stump neoplasia. Sparing of the cardinal ligament insertion provides support to the cervical stump. Hemorrhage and genitourinary complications are prevented by avoiding dissection of the parametrium at the level of endocervix. Comparison of data of the pelviscopic CISH procedure with the laparotomy approach in our preliminary series (n = 190) confirms our claims as to its safety. Data on long-term postoperative evaluation are ongoing, but the initial reports deny any postoperative discomfort. Larger randomized studies are required to prove its value compared with the existing techniques. We believe that with its multitude of benefits, the classic intrafascial serrated-edged macro-morcellated (SEMM) hysterectomy (= CISH) may emerge as an attractive alternative to conventional hysterectomy.

Adult↗
PubMed1995

First-pass metabolism of alcohol. Absence of diurnal variation and its inhibition by cimetidine after evening meal.

To determine whether the first-pass metabolism (FPM) of orally consumed alcohol varies with the time of day, 12 healthy male subjects were tested with both oral and intravenous alcohol (0.3 g/kg), in the morning and evening, always 1 hr after the same standard meal. The results revealed no significant differences in FPM (81.6 +/- 11.6 vs 92.8 +/- 10.6 mg/kg) or in any other index of alcohol absorption and metabolism. Eleven subjects were also tested in the evening after treatment with cimetidine, an H2-antagonist that inhibits gastric alcohol dehydrogenase activity in vitro. Compared to baseline, cimetidine (1 g/day for eight days) significantly decreased FPM (from 100.1 +/- 8.0 to 52.6 +/- 11.4 mg/kg, P < 0.01) and increased the systemic bioavailability of alcohol (from 66 +/- 3 to 82 +/- 4%, P < 0.01), as well as peak blood alcohol concentrations (from 4.3 +/- 0.4 to 5.9 +/- 0.5 mM, P < 0.05) and areas under the curve (from 5.1 +/- 0.5 to 7.0 +/- 0.5 mM/hr, P < 0.01). The results indicate the absence of diurnal variation in FPM and suggest that patients given cimetidine should be warned of its possible interaction with alcohol regardless of the time of day.

Administration, Oral↗
PubMed1995

Morphometric analysis of intestinal mucins under different dietary conditions and gut flora in rats.

Elucidation of the mechanisms that alter the biosynthesis, turnover, and degradation of intestinal mucins is relevant to the understanding of both the normal gut ecosystem and various intestinal diseases. In this study image analysis was used to quantify the effects of diet and microbial flora on the mucin composition of goblet and deep crypt cells, the number and volume density of mucin-containing cells, and the staining density of their stored mucins in the small and large intestine of germ-free and conventionally maintained rats fed two different diets. One was a coarsely ground commercial rodent diet containing crude fiber of cereal origin and the other a purified diet composed of finely powdered ingredients, including cellulose as a source of fiber. The changes in mucin production were also analyzed in germ-free rats colonized with a human flora. Feeding a commercial diet reduced the volume density of cells containing neutral and sulfomucins in the jejunum of conventional rats and the staining density of neutral and acidic mucins in the germ-free rats. Both rat and human floras reduced the number of cells containing acidic and sulfomucins and the staining density of neutral mucins in the small intestine of animals fed on a purified diet. However, inoculation of human flora increased the staining density of stored neutral and sulfated mucins in the cells of the large intestine. The results demonstrate that the dietary changes are influential in modifying the amount and proportion of mucins in the small intestine and the microbial flora in the large intestine.

Animal Feed↗