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Biomedical subjects

P K Basu

Publications and source records attributed to P K Basu.

At least 55 records · Page 3Linked to original sources

Toxic effects of topically administered Betagan, Betoptic, and Timoptic on regenerating corneal epithelium.

Re-epithelialization of 40 iatrogenic cornea ulcers produced in rabbits with iodine vapour burn were studied after topical administration of Betagan, Betoptic or Timoptic. At 24 hours there was no statistical difference in the wound healing rates among all the treated animal groups. At 48 hours, all the drug-treated groups were significantly different from the control group (p less than 0.05). There was no difference between the Betagan and Betoptic treated groups but the Timoptic-treated group was slower (p less than 0.05) than the Betoptic treated group, while the difference between the Timoptic and Betagan treated groups is not significant. At 72 hours the re-epithelialization rate of the Timoptic treated group was significantly slower than both the Betagan and Betoptic treated groups (p less than 0.05). By the 16th day following ulceration SEM examination revealed the Timoptic treated group had extensive epithelial cell desquamation, as well as hole formation and plasma membrane disruption. The Betagan treated corneas had mild superficial epithelial changes with loss of microvilli or slight desquamation. The Betoptic treated corneas were completely healed. In conclusion, Betagan is less toxic than Timoptic. Betoptic is the least toxic of the three drugs to regenerating rabbit corneal epithelium.

Administration, Topical↗

A morphometric study of endothelial cells of human corneas stored in MK media and warmed at 37 degrees C.

No study has yet been done to investigate the changes in endothelial cell size, perimeter, and density that may result from the warming of corneas in MK (McCarey-Kaufman) medium for specular microscopy. In the present investigation eye bank eyes were stored in MK medium at 4 degrees C and rewarmed daily for six days at 37 degrees C before specular photography of the endothelium was performed. These photographs were compared with wet mount preparations stained with trypan blue and alizarin red made from the same corneas and those stored without rewarming for six days. In addition all corneas were qualitatively analysed with the scanning electron microscope (SEM). The data from serial specular photography were insufficient to allow significant conclusions to be drawn about day to day changes in cell morphology. However, analysis of wet mount preparations revealed that cell density and perimeter varied significantly between those corneas rewarmed daily and those held in cold storage for six days. SEM studies showed an intact cell monolayer with cell loss along the folds of corneal endothelium. We therefore concluded that repeated rewarming at 37 degrees C of corneas stored in MK medium at 4 degrees has a deleterious effect on cell morphology and that folds induced by swelling of corneal tissue result in endothelial cell damage with some loss.

Cell Count↗

Phototoxicity of chlorpromazine on retinal pigment epithelial cells.

As it is known that chlorpromazine (CPZ) can bind to melanins as well as cause ocular phototoxicity, we investigated the cytotoxic effects of UV-visible irradiation of melanotic and amelanotic retinal pigment epithelial (RPE) cells in the presence of CPZ. At low concentrations (5 micrograms/ml) of CPZ a photosensitization reaction took place which lysed the cells as measured by the release of 51Cr from cells labelled with chromium. At concentrations of CPZ less than 5 micrograms/ml, no significant cell lysis occurred when the cells were incubated at 37 degrees C in the dark. As the concentration of CPZ was increased to 25 micrograms/ml or more, high percentages of cells were lysed. When the melanotic RPE cells were exposed to different concentrations of CPZ and grown in culture, the cell growth (multiplication) diminished drastically with low concentrations (less than 2 micrograms/ml CPZ). Vitamin E decreased the cell lysis both in the dark and upon irradiation. Oxygen radical scavengers such as glutathione, B-carotene, mannitol, D-penicillamine as well as superoxide dismutase and catalase did not decrease cell lysis. The phototoxic effects of CPZ was found not to be due to stable photoproducts formed during irradiation of CPZ.

Animals↗

Ultrastructural changes of the rabbit corneal epithelium and endothelium after Timoptic treatment.

The purpose of this study was to determine whether timolol maleate (Timoptic) is cytotoxic to the cornea. The drug was applied as topical drops for 1 month to control rabbit eyes and rabbit eyes from which the epithelium had been removed or to de-epithelialized full-thickness autografts. Clinically all corneas and grafts remained clear. Scanning and transmission electronmicroscopy however revealed is drug induced epithelial damage, defective epithelial wound healing, and endothelial changes. The endothelial changes were most prominent in the group of rabbits that underwent de-epithelialization before corneal grafting.

Animals↗

Changes of MK medium during storage of human cornea.

By comparing the composition of McCarey-Kaufman (MK) medium before and after corneal storage we attempted to identify specific physiological changes in the medium as predictors of tissue damage. We also tried to determine if hydrocortisone (a lysosomal membrane stabiliser) added to the medium could reduce tissue damage during storage. Corneas (human and rabbit) were stored in the MK medium with and without hydrocortisone for 4 days at 4 degrees C. The water and nitrogen contents of the stored cornea were compared with those of the fresh cornea. The medium was analysed before and after corneal storage to determine the concentrations of glucose, protein, and amino acids as well as pH and osmolarity. Scanning electron microscopy (SEM) was used to estimate the degree of the corneal endothelial cell damage. The nitrogen contents and dry weights of the steroid treated and untreated stored corneas were similar to those of the fresh unstored cornea. The steroid treated cornea contained a lesser amount of water than the untreated cornea. The cornea stored in medium without steroid took up a greater amount of glucose from the medium than the cornea stored in medium with steroid. As compared with their concentrations in the fresh unused medium the concentrations of leucine, lysine, and glycine were lower and that of glutamic acid was higher in both the media used for corneal storage. However, the steroid treated storage medium as compared with the untreated storage medium had a greater reduction in the lowering of leucine, lysine, and glycine, and a lesser reduction in the increase of glutamic acid. Steroid treated medium also had a lesser amount of protein released from the stored cornea. Changes in the pH and osmolarity of the media before and after corneal storage were not remarkable. SEM showed that the endothelial cells of the cornea stored in the medium containing steroid were less damaged than those of the cornea stored in the medium without steroid.

Amino Acids↗

Is there any difference in the photobiological properties of melanins isolated from human blue and brown eyes?

Investigations were carried out to determine whether the melanin present in the blue and brown eyes were eumelanin, the melanin present in black hair and dark skin, or pheomelanin, the melanin present in red hair and the skin of people with red hair. Our results showed that UV-visible irradiation of blue or brown eye melanin did not produce any superoxide. Irradiation of 51Cr-labelled Ehrlich ascites carcinoma cells in the presence of blue or brown eye melanin did not produce significant cell lysis. The electron spin resonance (ESR) signals of blue and brown eye melanins were very similar to those of eumelanin. Comparison of these findings with our previous results indicated that the blue and brown eye melanins are essentially eumelanin. The ESR signals further suggested that in the case of both blue and brown eye melanins the iris, ciliary body, choroid, and retinal pigment epithelium did not differ.

Aged↗

Ocular distribution of methyprylon after intravenous injection and transfer of the drug to recipient eye via corneal graft.

The purpose of the study was to determine chronologically the ocular distribution of methyprylon (Noludar) after a single intravenous injection (45 mg/kg) to rabbits, if the drug can be transferred from a donor treated with the drug to the recipient's eye via an allogeneic corneal graft, and the effect of methyprylon on corneal cell growth and protein synthesis. Gas chromatographic techniques showed that the drug was present in all the tissues and fluids of the eye from 0.5 to 18 hours after the injection. Postoperatively, the cornea and some of the other ocular tissues of the recipient contained methyprylon at least 24 hours after transplantation. Clinically, the graft remained clear, and there was no adverse reaction in the recipient eye for an observation period of 24 hours. Tissue culture studies showed that about 10 times more methyprylon than was found in the donor cornea was necessary to produce a cytotoxic reaction on the corneal cells in terms of cellular growth rate and protein synthesis. Our animal experiments and cytotoxicity tests suggest that corneas of donors treated with a high dose of methyprylon would be suitable for use as grafts.

Animals↗

Inhibition of vascular endothelial cell growth and trypsin activity by vitreous.

Vitreous from bovine, human and chick embryo has been found to contain a trypsin inhibitory activity. Chymotrypsin-inhibitory activity was also identified in bovine and chick embryo vitreous. Following either ultrafiltration or Bio Gel P-10 chromatography, these activities appear in fractions having a molecular weight greater than 10000 MW (ultrafiltration) or greater than 13000 MW (P-10 void volume), and are separable from low molecular weight aortic endothelial cell growth inhibitory activity present either in the ultrafiltrate or P-10 retarded volume. Treatment of the trypsin inhibitory fraction with hyaluronidase had no effect on trypsin inhibition, nor did addition of hyaluronic acid inhibit trypsin. Chick embryo vitreous and hyalocyte-conditioned medium were found to contain aortic endothelial cell growth inhibitory activity in both the void volume and retarded volume fractions following Bio Gel P-10 chromatography. Both the 6200 MW bovine vitreous endothelial cell growth inhibitor and the high molecular weight chick embryo vitreous endothelial cell growth inhibitor (greater than 13000 MW) were similar, in that most of the activity did not bind to heparin linked to Sepharose CL-6B.

Animals↗

The BB-rat--an authentic model of human diabetic retinopathy.

The pathogenesis of diabetic retinopathy has not been fully explained. The earliest histological lesion is the loss of intramural pericytes and thickening of the basement membrane. Increased activity of the polyol pathway is a probable mechanism for these two abnormalities. Investigations have suffered from the lack of an exact animal model simulating the human condition. Examination of the retina in the spontaneously diabetic BB-rat demonstrated degeneration and loss of intramural pericytes, a progressive increase in basement membrane thickness, and microinfarctions with areas of non-perfusion. Therefore, this model may be used to clarify the biochemical mechanism(s) linking the metabolic abnormalities of diabetes and the retinopathy.

Animals↗

Vascular endothelial cell growth inhibitor of normal and pathologic human vitreous.

Normal and pathologic human vitreous have been analyzed for the presence of a low-molecular weight inhibitor of aortic endothelial cell proliferation. Vitreous was subjected to gel chromatography and the material appearing in the retarded volume (less than 13,000 daltons) was tested for its ability to inhibit tritiated thymidine incorporation into DNA by calf aortic endothelial cells. Depending on the sample of vitreous analyzed, one or more fractions showing inhibitory activity were identified in each case.

Adult↗

Should corneas from donors receiving a high dose of salicylate be used as grafts: an animal experimentation.

Rabbits were injected intravenously with a single dose of sodium salicylate (350 mg kg-1) in order to study the following in vivo: (i) the distribution of the drug in the various eye tissues and fluids, particularly the cornea and aqueous humour, and (ii) transfer of the drug from corneal grafts obtained from donors receiving sodium salicylate to the eye tissues of the recipient. In additional experiments, the in vitro effects of sodium salicylate on the growth and protein synthesis of corneal cells grown in tissue culture were also studied. In vivo experiments showed that the periods during which we observed the highest concentration of salicylate in the serum and in the eye tissues were within 30 min and 2 hr respectively following the injection. These experiments also showed that salicylate was transferred from the treated donor via corneal graft to the recipient's eye tissues where it could still be detected 48 hr after the operation. In vitro experiments showed that a 50% inhibition of cell growth was obtained at a concentration of about 1000 micrograms salicylate ml-1 while protein synthesis was decreased by 50% at a concentration of about 200 micrograms ml-1. A consideration of our data from the in vivo and in vitro experiments together suggests that ingestion by the donor of high concentrations of salicylate may have the potentiality of subjecting the corneal endothelial cells to cytotoxic concentrations of the drug thus jeopardizing the success rate of corneal graft operations.

Animals↗