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Biomedical subjects

P Ghosh

Publications and source records attributed to P Ghosh.

At least 289 records · Page 16Linked to original sources
PubMed1989

Impact of nonlethal levels of Metacid-50 and carbaryl on thyroid function and cholinergic system of Channa punctatus.

With regard to brain acetylcholinesterase and acetylcholine and serum triiodothyronine (T3) and thyroxine (T4) profiles, a biphasic response pattern was elicited in Channa punctatus chronically exposed to nonlethal doses of locally used pesticides, namely, Metacid-50 and Carbaryl. Data revealed that these xenobiotics caused significant inhibition of brain acetylcholinesterase activity and a decrease in thyroxine level accompanied by a concurrent increase in acetylcholine accumulation and T3 level. It is surmised that Metacid-50 and Carbaryl influence both neural and hormonal functions.

Acetylcholine↗
PubMed1988

The quantitative discrimination of corneal type I, but not skeletal type II, keratan sulfate in glycosaminoglycan mixtures by using a combination of dimethylmethylene blue and endo-beta-D-galactosidase digestion.

The quantitation of individual glycosaminoglycans in mixtures of polyanions using the dimethylmethylene blue (DMB) method described by R. W. Farndale, D. J. Buttle, and A. J. Barrett (1986, Biochim. Biophys. Acta 883, 173) is dependent on enzymatic hydrolysis by specific polysaccharidases. While using this method to examine the keratan sulfate (KS) of the intervertebral disc we found that digestion with commercially available keratanase decreased binding to DMB by less than 30%, whereas corneal KS was reduced by 85%. However, by preincubating the KS fractions with endo-beta-D-galactosidase prior to keratanase treatment the corneal KS could be completely digested and disc KS digestion increased to 60%. It is suggested that the resistance of the disc KS to these digestive procedures arises from branching and/or sites of multisulfation on the polysaccharide chain. Agarose gel electrophoresis and compositional analyses of the keratan sulfates supported such an interpretation.

Chondroitin Sulfate Proteoglycans↗
PubMed1988

Cysteine proteinase inhibitors of the canine intervertebral disc.

Several species of cysteine proteinase inhibitors have been demonstrated in the greyhound intervertebral disc which were resolved into four species (Mr 15,800, 16,600, 17,200 and 17,800) by gelatin-SDS-polyacrylamide gel electrophoresis. Reductive alkylation did not affect their inhibitory capability nor their electrophoretic migration on gelatin-SDS-polyacrylamide gel electrophoresis. The cysteine proteinase inhibitors from the nucleus pulposus and annulus fibrosus were identical as assessed by the aforementioned criteria, although the level in the nucleus was found to be higher than that in the annulus. Ion-exchange chromatography demonstrated distinct acidic and basic forms of the disc cysteine proteinase inhibitor. The latter species was the most abundant and its Mr was determined to be 16,900 by gelatin-SDS-polyacrylamide gel electrophoresis. Both forms were shown to be strongly inhibitory against the cysteine proteinases, papain and ficin, but were less strongly inhibitory against cathepsin B (EC 3.4.22.1). Presumably these disc cysteine proteinase inhibitors play a regulatory role in the metabolism of proteoglycans and collagen by endogenous cysteine proteinases.

Animals↗
PubMed1988

Anti-rheumatic drugs and cartilage.

In this chapter an attempt has been made to draw together the known biology of cartilage and some of the mechanisms thought to be responsible for its failure in arthritis. The picture is far from complete but we are now in a good position to use this information to help appraise the pros and cons of the wide range of drugs now available to treat articular disorders. For convenience, these drugs were classified as NSAIDs, corticosteroids and chondroprotective agents. The influence of each of these classes on the metabolism of cartilage was examined in the light of published laboratory and clinical studies. It has been clearly shown that not all NSAIDs are the same. While many of the older drugs provided no benefit to cartilage metabolism, and in some instances suppressed it, the more recently discovered molecules appear to be free of these undesirable effects. Tiaprofenic acid, diclofenac and piroxicam emerged as drugs with little or no harmful effects on cartilage metabolism when used at concentrations within the human therapeutic range. For all NSAIDs, their potential effects on cartilage must be weighed against their respective anti-inflammatory potency, half-life, and effects on the gastric mucosa and other tissues. Other chapters in this book have addressed these important problems. The long-acting corticosteroids, betamethasone and triamcinolone hexacetonide, also appear to offer some benefit in the management of OA; however, as in RA, their use should be restricted to short-term applications. In terms of cartilage metabolism the chondroprotective agents pentosan polysulphate, Arteparon and Rumalon have been the most extensively studied class of drugs. While the laboratory studies have provided convincing evidence of their chondroprotective efficacy, it has been difficult to prove this clinically. This dichotomy of opinion (laboratory versus clinical) stems largely from the inadequacy of the methodologies currently available for the objective clinical assessment of patient response to anti-rheumatic drug therapy. With the advent of nuclear magnetic resonance imaging techniques and monoclonal antibodies to detect specific proteoglycan breakdown fragments in synovial fluid and plasma, the prospects for a unified research approach for the evaluation of these agents may now be possible.

Animals↗
PubMed1988

Acquired tracheo-oesophageal fistula (a case report).

A case of acquired tracheo-oesophageal fistula in a 16-year-old boy, caused by a bullet wound, is presented. The fistula was located just below the cricoid cartilage and was successfully treated by excision of the fistula followed by interposition of the sternomastoid muscle.

Adolescent↗
PubMed1988

Binding of haptoglobin, inter-alpha-trypsin inhibitor, and alpha 1 proteinase inhibitor to synovial fluid hyaluronate and the influence of these proteins on its degradation by oxygen derived free radicals.

Synovial fluid from 201 normal and pathological knee joints was subjected to gel filtration by Sepharose CL-2B chromatography to separate hyaluronic acid (HA) from unbound proteins, which were retarded on this column. HA from all normal fluids was excluded from the gel and contained 1% or less bound protein. Synovial fluids taken from joints of patients with rheumatoid arthritis (RA) contained considerably more protein bound to HA. In 46% of RA samples the level of protein was greater than 4%, whereas only one fluid examined from osteoarthritic joints contained this amount. The proteins bound to HA from RA joints were identified by sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS-PAGE) and immunodiffusion techniques as the acute phase proteins alpha 1 proteinase inhibitor, inter-alpha-trypsin inhibitor, and haptoglobin. The average relative percentages of these proteins bound to HA were 17.6%, 32.6%, and 29.2% respectively. These HA-protein complexes could be generated in vitro by mixing normal (low protein) HA with any one of the three acute phase proteins. The HA-protein complexes formed in vitro with inter-alpha-trypsin inhibitor or haptoglobin, and those isolated from RA synovial fluids, were more resistant to degradation by oxygen derived free radicals (ODFR) than HA from normal fluids. From these findings we conclude that certain acute phase proteins diffusing into synovial fluid during inflammatory episodes may play an important part in protecting HA from depolymerisation by activated phagocytes.

Alpha-Globulins↗
PubMed1988

The role of plasminogen in interleukin-1 mediated cartilage degradation.

Explants of 35SO4 labelled rabbit articular cartilage, cultured for 3 days with either 5 X 10(4) rabbit peritoneal cells (PEC) or 1:10 macrophage conditioned medium (MCM), released 30-40% of labelled proteoglycans into the medium while controls released 8-12%. The addition of 1 mM 4 aminophenylmercuric acetate (APMA) or 0.2 U/ml plasminogen increased proteoglycan release to 85%. Similar results were obtained when recombinant human interleukin-1 (IL-1) was used instead of MCM. Further, supernatant from MCM stimulated chondrocytes, incubated with dead cartilage explants for 3 days, did not significantly increase proteoglycan release above the background level of cartilage alone (7-10%), nor did the addition of 5 X 10(4) PEC to cultures of dead cartilage explants plus supernatant from MCM stimulated chondrocytes make any significant difference, indicating that supernatant from MCM stimulated chondrocytes and PEC alone had negligible cartilage proteoglycan degrading activity in these experiments. The inclusion of 0.1 mM APMA or 0.2 U/ml plasminogen in cultures of dead cartilage explants plus supernatant from MCM stimulated chondrocytes, however, increased proteoglycan release to 80-93%, with or without PEC. Our results suggest that plasminogen, activated by a product from IL-1 stimulated chondrocytes, greatly enhanced IL-1 mediated cartilage degradation by activating latent metalloproteinases.

Animals↗
PubMed1987

Effects of gentamicin on sphingomyelinase activity in cultured human renal proximal tubular cells.

We have previously shown that cultured human proximal tubular cells (PT) incubated with gentamicin contain numerous "myeloid bodies." This morphological change was accompanied by the storage of phosphatidylcholine and sphingomyelin. In order to delineate the biochemical mechanisms responsible for the accumulation of sphingomyelin in cells incubated with gentamicin, we pursued detailed studies on the activity of sphingomyelinase. Characterization studies on sphingomyelinase revealed that this enzyme has a bimodal pH optima in PT cells. Optimum activity was observed at pH 5.6 (designated as acid sphingomyelinase, A-SMase) and at pH 7.4 (designated as neutral sphingomyelinase, N-SMase). The activity of both the enzymes increased proportionately in control cells as a function of days of incubation. The activity of A-SMase was 16% lower in cells incubated with gentamicin as compared to control. The most striking observation was a gradual decline in the activity of N-SMase in cells incubated with gentamicin. Thus, following 21 days of incubation of cells with 0.3 mM gentamicin, the N-SMase was 2.7-fold lower than control cells. Mg2+ stimulated and Triton X-100 inhibited the activity of N-SMase. Whereas Mg2+ had no effects, Triton X-100 stimulated the activity of the A-SMase in PT cells. Moreover, A-SMase was relatively more heat-resistant than the N-SMase. The Km values for sphingomyelin using A-SMase in control cells and cells incubated with gentamicin were 0.07 X and 0.016 X 10(-7) M, respectively, whereas the Km values for sphingomyelin using N-SMase in control cells and cells incubated with gentamicin were 1.8 X and 1.5 X 10(-7) M, respectively. These findings suggest that gentamicin exerts a competitive inhibition of the A-SMase in PT cells. In contrast, gentamicin exerts a noncompetitive inhibition of the N-SMase in PT cells. Subcellular fractionation studies revealed that A-SMase was exclusively localized in the "lysosome-rich" fraction, whereas most, if not all, the N-SMase was localized in the microsomal fraction and "plasma-membrane"-rich fraction in cultured PT cells. Cells incubated with gentamicin for 21 days contained 25% lower activity of A-SMase associated with the lysosomal fraction as compared to control. In contrast, N-SMase activity in the microsomal and plasma membrane fraction was one-half as compared to control. We conclude that gentamicin-mediated decrease in sphingomyelinase activity may be responsible for the storage of sphingomyelin in cultured human PT cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Cells, Cultured↗
PubMed1987

Neutral proteinases of the human intervertebral disc.

Disc tissue consisting of pooled annuli fibrosus and nuclei pulposus from the cadaver of an adolescent aged 19 years was extracted with 4.0 M Gu-HCl. Proteins of low buoyant density (p less than or equal to 1.38 g/ml) containing the disc enzymes and inhibitors were separated from proteoglycans of high buoyant density (p greater than or equal to 1.50 g/ml) by density gradient ultracentrifugation. Sephadex G-75F gel chromatography followed by trypsin affinity chromatography was then used to resolve disc proteolytic and trypsin inhibitory activities. The results obtained were strongly suggestive of the presence of a high molecular weight zymogen which upon activation generated a population of smaller molecular weight proteinases. The disc proteinases obtained by this process showed similar properties in terms of: their pH optima (7.4-7.6); their inhibition patterns by class-specific proteinase inhibitors; their variation of activity as a function of NaCl and lysine concentrations; and the hydrodynamic size of their proteoglycan degradation products. The activated disc neutral proteinase demonstrated many characteristics in common with plasmin; however, unlike the latter, the disc proteinases also showed some calcium dependence.

Adolescent↗
PubMed1987

Systemic administration of glycosaminoglycan polysulphate (arteparon) provides partial protection of articular cartilage from damage produced by meniscectomy in the canine.

Bilateral medial meniscectomy was undertaken in 14 mature beagles. Another two underwent arthrotomy (sham-operated controls). One week after surgery, six of the meniscectomised animals were administered glycosaminoglycan polysulphate (Arteparon) (2 mg/kg) subcutaneously three times a week for 3 weeks, then twice weekly until killed 23 weeks later. Two months before death all animals were given Na2(35)SO4 (1.0 mCi/kg) intravenously. At autopsy, articular cartilage (AC) from the medial and lateral compartments, as well as from the femoral trochlear groove and femoral head, was sampled. Proteoglycans (PGs) were isolated by 4.0 M Guanidine hydrochloride (GuHCl) extraction of AC and purified by ultracentrifugation. The PG monomers were assayed for hexuronic acid, protein, and hexosamines (galactosamine/glucosamine), and their ability to aggregate. The results indicated that Arteparon provided some protective effect to AC in the meniscectomised compartment as demonstrated histologically by reduced surface fibrillation, diminished chondrocyte cloning, and maintenance of alcianophilia. The levels of PGs and hexuronate-protein ratios in medial AC of drug-treated meniscectomised animals were found to be comparable to sham controls, whereas these parameters in the nondrug-treated meniscectomized group were depressed.

Animals↗
PubMed1987

The response of the canine intervertebral disc to immobilization produced by spinal arthrodesis is dependent on constitutional factors.

Posterior lumbar spinal fusion was performed on five mature greyhounds. Two months prior to death, all of the surgical and five age-matched control greyhounds were given Na2(35)SO4 (1.0 mCi/kg) intravenously. All fusion animals were killed 6 months postoperation, and discs beneath the fusion mass as well as those adjacent to it (parafusion discs) were sampled separately and dissected into the nuclei pulposi and annuli fibrosi (AF). Proteoglycans (PGs) were extracted with 4.0 M GuHCl and then purified by CsCl density gradient ultracentrifugation. These PG monomers were subjected to Sepharose CL-2B chromatography, and their hydrodynamic size and ability to aggregate were determined. The level, extractability, and hydrodynamic size of PGs in the AF of fusion discs were found to be greater than those in control discs, as were the keratan sulfate core protein complexes prepared by chondroitin ABC lyase digestion. The ability of the 60-day-old PG subunit populations, isolated from fusion discs, to aggregate was also higher than controls. There was, however, no difference between the galactosamine/glucosamine, galactosamine/protein, glucosamine/protein, or hexuronate/protein ratios of PGs in fusion and control discs.

Animals↗
PubMed1987

The synthesis of hyaluronic acid by human synovial fibroblasts is influenced by the nature of the hyaluronate in the extracellular environment.

Various cell lines of human synovial fibroblasts derived from synovium obtained at the time of biopsy or total joint-replacement surgery have been established. The synthesis of 3H-labelled hyaluronic acid (HA) in these cells has been determined, and the effects of adding HA of varying molecular size to the cultured cells examined. The results obtained clearly show that the in vitro synthesis of HA by these cells is influenced by the concentration and molecular weight (MW) of the HA in their extracellular environment. Synovial fibroblasts derived from an osteoarthritic joint demonstrated the most marked response on exposure to exogenous HA, showing a stimulation of HA synthesis with preparations of weight-average molecular weight (Mw) greater than 5 X 10(5) in a concentration dependent manner. HA preparations with Mw less than 5 X 10(5) showed little or no effect except at high concentrations where a suppression of biosynthesis was observed. A model to explain these findings is proposed.

Adult↗
PubMed1987

A new mycotoxin from Aspergillus candidus Link isolated from rough rice.

A new mycotoxin (AcT1) was obtained from the mycelium of Aspergillus candidus Link isolated from rough rice stored under tropical conditions. AcT1 with the mol-formula C28H40O showed bright greenish blue fluorescence under uv (254 nm). The spectral and other characteristics indicated that the compound was a new one. The LD50 of the toxin on white rats was found to be 4.5 mg/Kg when injected intraperitoneally.

Animals↗