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Biomedical subjects

N Agnihotri

Publications and source records attributed to N Agnihotri.

At least 19 recordsLinked to original sources

Communication of radiation-induced stress or bystander signals between fish in vivo.

We report data in this paper suggesting that fish irradiated to 0.5 Gy total body dose can release factors into the water that signal other unexposed fish and cause induction of bystander effects expressed as increased cell death in a reporter system. Radiation-induced bystander effects, resulting in the appearance of radiation damage or induction of typical radiation responses in unirradiated cells and tissues are now an established consequence of exposure to low doses of ionizing radiation, however little work has been done in vivo or in species other than humans or mice. In these experiments rainbow trout were irradiated and then paired with unirradiated fish for two hours. Additionally, unirradiated fish were placed in water which had previously been used to hold irradiated fish for 2 h. Sham-irradiated fish and absolute control fish were also examined all using blind protocols. Following a two h incubation period, at these various exposure regimes, the fish were killed by a blow to the head and dissected. Five organs were removed from each fish and tissue explants were cultured using an established technique. After 2 days, the culture medium was harvested and used in a reporter assay to determine whether a bystander effect had been induced. The explants were cultured on in Clonetics growth medium for a further 14 days then fixed for assay of radiation response proteins. The responses varied according to the cell type in the original explants, with the gill and fin showing the most pronounced response. The results suggest that communication signals leading to a typical radiation response can be passed between fish and seem to involve secretion of a chemical messenger into the water.

Animals↗

Aerobic bacterial isolates from burn wound infections and their antibiograms--a five-year study.

A retrospective study of major aerobic bacterial isolates from pus/wound swabs taken from patients admitted to the burn unit at Govt. Medical College Hospital, Chandigarh, India, over a period of 5 years (June 1997-May 2002) was undertaken. The study was carried out to determine the bacterial profile and antimicrobial susceptibility of the isolates and to describe the change in trends over the study period. The pus/wound swabs yielded very high culture positivity (96%) for 665 total isolates. Pseudomonas aeruginosa was found to be most common isolate (59%) followed by Staphylococcus aureus (17.9%), Acinetobacter spp. (7.2%), Klebsiella spp. (3.9%), Enterobacter spp. (3.9%), Proteus spp. (3.3%) and others (4.8%). Although P. aeruginosa continued to remain the predominant isolate over the five years, a constant and significant increase in the incidence of Acinetobacter spp. was found. Amikacin was found to be the most effective drug against gram negative bacteria, however, resistance to it was significantly increased over 5 years. For S. aureus and P. aeruginosa netilmicin and piperacillin were found to be the most effective drugs. Most of the isolates showed high level resistance to antimicrobial agents.

Bacteria, Aerobic↗

Activity of brush border membrane enzymes in proximal jejunum of portal hypertensive rats.

PURPOSE: Malabsorption accompanies portal hypertension, especially when associated with chronic liver disease. The development of portal hypertension is accompanied by significant alterations in the splanchnic microcirculation. In this study, the effect of extrahepatic and intrahepatic portal hypertension on brush border membrane enzymes was estimated. METHODS: Portal hypertension was induced in rats by partial portal vein ligation (PPVL) (n = 6) and common bile duct ligation (CBDL) (n = 6), and the activity of sucrase, lactase, alkaline phosphatase, and leucine aminopeptidase (LAP) in the intestinal homogenate was measured. RESULTS: Intrasplenic pulp pressure (ISPP) (in cm of saline) was found to be elevated in PPVL (21.3 +/- 1.47) and CBDL animals (21.5 +/- 1.79) as compared with findings in their respective sham-operated controls (12.74 +/- 0.86, 11.83 +/- 1.04). Only sucrase and LAP activity was significantly elevated (P < 0.05) in the PPVL group. No changes were observed in the CBDL group. CONCLUSION: Only sucrase and LAP activities were increased in PPVL rats.

Animals↗

Inhibition of Helicobacter pylori adherence by a peptide derived from neuraminyl lactose binding adhesin.

Helicobacterpylori, like many other gut colonizing bacteria, binds to sialic acid rich macromolecules present on the gastric epithelium. NLBH (neuraminyl lactose binding haemagglutinin) a 32 kDa adhesin located on the surface of H. pylori has been shown to have specific affinity towards NeuAcalpha2,3Galbeta1,4Gluc(3'SL). This sialic acid moiety is over-expressed in an atrophic stomach undergoing parietal cell depletion. Antibodies against a lysine rich peptide fragment of NLBH inhibit agglutination of human erythrocytes. This lysine rich sequence from NLBH was proposed to be the receptor-binding site. In order to elucidate the binding of NLBH to gastric epithelium, a peptide (D-P-K-R-T-I-Q-K-K-S) was synthesized. A series of experiments were performed involving adherence inhibition assays, 2D-NMR, molecular modelling and measurement of modulation in acid secretion. Results indicated that the peptide fragment could be involved in receptor recognition, which is important for the binding of H. pylori to gastric epithelium. The binding is possibly through hydrogen bonding. Two lysines and a threonine residue seem to be within the hydrogen bonding distance of NeuAcalpha2,3Galbeta1,4Gluc. Further, in vitro assays were performed to evaluate the role of the peptide on acid secretion by parietal cells isolated from human fundal biopsies. Interestingly, the peptide increases acid secretion only in H. pylori negative and in treated patients but not in H. pylori positive patients. This highlights the role of NLBH in acid secretion and could be of some consequence in the prognosis of the disease.

Animals↗

Modulation of acid secretion in common bile duct ligation-related gastropathy in Wistar rats.

BACKGROUND: Portal hypertensive gastropathy is associated with fundic gland atrophy, resulting in a decrease in chief and parietal cells, and diminished acid secretion. METHODS: Acid secretion by isolated parietal cells was measured (acridine orange retention), along with the levels of various second messengers (intracellular Ca(2+), cyclic adenosine monophosphate and protein kinase C) in the common bile duct, ligated portal hypertensive rats and compared with sham-operated controls. RESULTS: There was a significant decrease in the response of isolated parietal cells to the secretagogues histamine and carbachol. This resulted in the blunted acid secretion in the common bile duct ligated group. In addition, all the second messengers studied were significantly decreased as compared with the sham-operated controls. CONCLUSION: These results suggest that the blunted acid secretory response in the portal hypertensive rat is caused by an alteration in the intracellular signal transduction mechanism.

Acridine Orange↗

Micro-perfusion flow cell for imaging cultured cells.

We present a unique design for a flow cell with a small working volume that allows rapid displacement of media viewed under high power and short working distance objectives. The flow cell has a small internal depth (ca. 0.033 cm) and volume (ca. 0.05 mL) and is easy to handle. Made of Delrin, the flow cell is biologically inert. We have used the flow cell for fluorescence imaging of PC12 cells loaded with tetramethylrhodamine dextran (TMRD) and other dyes.

Animals↗

Extrahepatic portal hypertensive gastropathy in Wistar rats: modulation of acid secretion in isolated parietal cells.

Stimulated acid secretion in portal hypertensive gastropathy is blunted and could be due to defective signal transduction in the parietal cell. Therefore, an attempt was made to study the levels of second messengers in parietal cells in experimental extrahepatic portal hypertensive gastropathy. Our aim was to measure acid secretion, intracellular free calcium, calcium transport, cyclic AMP, and ATP levels in the parietal cells isolated from the gastric mucosa of portal hypertensive rats. Acid secretion using acridine orange, intracellular free calcium using Fura-2/AM, calcium influx and efflux by 45CaCl2 and cyclic AMP by RIA kits were measured in unstimulated and histamine- and carbachol-stimulated isolated parietal cells in rats with partial portal vein ligation and sham operation. ATP was measured by HPLC. In portal hypertensive gastropathy, stimulated acid secretion was blunted, and there was a decrease in basal intracellular free calcium. Calcium influx and efflux were at a higher level, and there was a decrease in elevation of intracellular free calcium and cyclic AMP levels with secretagogues. There was also a decrease in ATP. In conclusion, there exists a low energy state in addition to multiple aberrations at the second messenger level in parietal cells in portal hypertensive gastropathy.

Adenosine Triphosphate↗

Characterization of lymphocytic subsets and cytokine production in gastric biopsy samples from Helicobacter pylori patients.

BACKGROUND: This study characterized the phenotypic subsets of isolated gastric lymphocytes and the cellular immune response in cultured gastric biopsy specimens. METHODS: Endoscopy specimens from 40 Helicobacter pylori-positive and 40 H. pylori-negative patients were studied. a) Isolated gastric lymphocytes were analysed for CD4+, CD8+ T-lymphocyte subsets, activated T cells, and natural killer cells on a fluorescence-activated cell sorter, using monoclonal antibodies. b) The supernatant of cultured gastric biopsy specimens were assayed for interleukin (IL)-2, IL-4, and IL-6 levels. RESULTS: In H. pylori-positive patients there was (a) a decrease in CD4+/CD8+ T cells, no change in activated T cells, and an increase in natural killer cells, and (b) no change in IL-2 levels and a significant increase in IL-4 and IL-6 levels. CONCLUSIONS: There is an increase in CD8+ lymphocytes and natural killer cells, and the observed increase in IL-4 and IL-6 might be important in H. pylori-associated gastritis.

Adult↗

Morphological changes associated with long-term potentiation.

Long-term potentiation (LTP) is a long-lasting form of synaptic plasticity induced by brief repetitive afferent stimulation that is thought to be associated with learning and memory. It is most commonly studied in the hippocampus where it may last for several weeks, and involves the synthesis of new proteins that might play a structural role. In this review we summarize the evidence in favor of modifications of neuronal architecture during LTP. We focus our attention on changes occurring at the level of single synapses, including components of postsynaptic dendrites (dendritic spines, the postsynaptic density, and synaptic curvature), of presynaptic terminals, and the formation of new synapses. We conclude that although many morphological changes at various sites have been observed during LTP, there is no definitive proof in favor of structural changes associated with LTP. However, morphological modifications remain a valid candidate for mechanisms of learning and memory.

Animals↗

Free cyclic AMP increases in PC12 cells on depolarization.

The temporal dynamics of the intracellular second messenger cyclic AMP (cAMP) were monitored in living PC12 cells by digital fluorescence ratio imaging using FlCRh, a single-excitation dual-emission cAMP indicator. When the cells were depolarized by exposure to high K+, the free cAMP concentration was elevated, and then slowly decreased back to resting levels when the depolarizing stimulus was removed. Furthermore, the cAMP elevation due to depolarization decreased with successive depolarizations.

Action Potentials↗

Diminution in parietal cell number in experimental portal hypertensive gastropathy.

Portal hypertensive gastropathy is a well-established clinical entity. Although stimulated acid secretion has been found to be decreased in portal hypertensive rats, the parietal cell mass has not been studied. Portal hypertension was produced in Wistar rats either by partial portal vein ligation (N = 16) or by common bile duct ligation (N = 23) and confirmed by intrasplenic pulp pressure measurement. The parietal cells were isolated and counted in a Neubaur hemacytometer. The parietal cell count was also done in microscopic sections at direct histopathological examination. The viable, isolated parietal cell count and parietal cell count on histopathological examination were significantly decreased in partial portal vein ligated rats. Similarly, in common bile duct ligated rats, the parietal cell count was decreased as compared to sham-operated rats. In experimental portal hypertensive gastropathy there is a decrease in parietal cell number.

Animals↗

Experimental extrahepatic obstruction of portal vein: documentation of histopathological alterations in liver and extrahepatic tissues.

The histopathology of the portal vein in human extrahepatic portal vein obstruction (EHPVO) in the form of cavernomatous transformation of the portal vein is well documented. However, a similar detailed histopathology has not been documented in experimental EHPVO, even though the model has been in existence for many years. An experimental model of EHPVO was created by partial ligation of the portal vein in 20 rats. A control group of 11 sham operated (SO) rats was also studied. Development of portal hypertension was confirmed by measurement of intrasplenic pulp pressure (19.7 + 7.1 cm normal saline vs 12.9 + 1.1 cm of normal saline in SO rats). Histopathological evidence included varices in the oesophagus and congestion in the submucosa and mucosa of the stomach and capillarization of sinusoids in the spleen. Animals were killed at 22-265 days post-ligation. While SO animals showed mild focal sinusoidal congestion and a normal portal vein, the experimental group of animals with EHPVO showed oedema around the portal vein, multiple dilated vascular channels near the hilum resembling cavernomatous transformation and severe sinusoidal congestion. Jejunum and kidneys showed congested veins in the experimental group. None of the SO animals showed this change. Thus experimental EHPVO does reproduce some histological changes observed in human EHPVO.

Animals↗

Plasma reactive nitrogen intermediate levels in patients with clinically active rheumatoid arthritis.

We studied reactive nitrogen intermediate levels in 31 patients with active rheumatoid arthritis (RA) taking indomethacin and 20 healthy controls using nitrite and citrulline levels, measured by spectrophotometry, as markers. Twenty patients with RA were followed up after 4 and 8 wk of treatment with additional therapy in the form of methotrexate. Mean nitrite levels in 31 patients were 0.94 +/- 0.41 mumol/ml and 20 controls it was 1.18 +/- 0.99. After treatment with methotrexate for 4 and 8 wk the levels were 0.9 +/- 0.45 and 1.25 +/- 1.15 mumol/ml, respectively. Mean citrulline levels in all patients was 1.68 +/- 0.11 and controls was 1.39 +/- 0.6 mumol/ml. Following therapy with methotrexate for 4 and 8 wk the levels were 1.40 +/- 0.49 and 1.40 +/- 0.51 mumol/ml, respectively. It is possible that serum levels of these products may not reflect alterations in the synovial fluid levels. Alternatively, whatever lowering may have been achieved by the anti-inflammatory effect of the therapy may have been countered by drug derived free radicals.

Adult↗

A flowcytometric study of CD4+ & CD19+ cell lymphopenia in patients with alopecia areata.

Twenty five patients with alopecia areata and 20 healthy controls were studied by flowcytometry employing direct two colour immunofluorescence in erythrocyte lysed whole blood. A significant reduction was observed in helper/inducer (CD4+) lymphocytes (29.4 +/- 7.4 vs 39.45 +/- 8.0, P < 0.01) and B (CD 19+) lymphocytes (11.04 +/- 6.57 vs 15.0 +/- 5.05, P < 0.01) in comparison with healthy controls. Decrease in T-lymphocytes (CD3+) and suppressor/cytotoxic (CD8+) was not significant and activated (HLADR+) lymphocytes and natural killer cells (CD16+ and CD56+) were within normal limits. Our findings suggest a significant T helper (CD4+) and B (CD19+) cell lymphopenia in alopecia areata.

Adolescent↗

Duodenal ulcer. Calcium status in isolated parietal cells.

Although the etiology of duodenal ulcer is not known, its treatment with drugs that reduce acid secretion is well accepted. The central role of calcium in stimulus-secretion coupling resulting in acid secretion by gastric parietal cells is documented. However, the status of intracellular calcium in gastric parietal cells in the basal state in patients with duodenal ulcer is not known. Multiple endoscopic gastric mucosal biopsies from the corpus of the stomach of 52 patients were processed and isolated parietal cells were studied. Intracellular calcium was estimated using fura-2-acetoxymethyl ester. Influx and efflux were determined by using radioactive calcium. Acridine orange retention was used to assess acid production. Only calcium influx at 20 min was significantly (P < 0.01) more in patients with duodenal ulcer as compared to the control group. There was no difference between the groups in calcium influx at 0 and 60 min; calcium efflux at 0, 20, and 60 min; intracellular free calcium and acid secretion. We conclude that in the unstimulated state calcium homeostasis in isolated parietal cells of patients with duodenal ulcer shows only a minimal difference as compared to controls.

Adult↗

Role of reactive oxygen species in renal damage in experimental leprosy.

Renal involvement is known to occur in leprosy. In the present study the possible role of reactive oxygen species (ROS) in causation of renal damage in mice infected with Mycobacterium leprae has been investigated. At least six animals from each group (control and infected) were killed at 0 day, 3, 6 and 9 months postinfection. The results showed a significant increase in the chemiluminescence (CL) response of peritoneal macrophages which was maximum between 3 and 6 months. No significant increase was observed in CL response of blood neutrophils. A significant increase in lipid peroxidation was observed at 3 and 6 months as evident by an increase in malondialdehyde levels. The increased ROS production might be the cause of lipid peroxidation. The renal damage is alos evident by decrease in the activity of renal brush border membrane enzymes, namely, alkaline phosphatase, leucine aminopeptidase and r-glutamyl transpeptidase. Thus ROS might play a role during early stages of M. leprae infection but in the later stages other immunological mechanisms may overpower the effect of ROS.

Alkaline Phosphatase↗