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Biomedical subjects

K Yee

Publications and source records attributed to K Yee.

10 recordsLinked to original sources

The role of thyroid hormone in the regulation of hepatic carbamyl phosphate synthetase activity in Rana catesbeiana.

Both spontaneous and thyroid hormone (TH)-induced metamorphosis of Rana catesbeiana are accompanied by a marked increase in the activity of the urea cycle enzyme carbamyl phosphate synthetase (CPS). The increase induced by exogenous TH is de novo synthesis of enzyme and appears to be secondary to an increase in the CPS mRNA level resulting from the elevated plasma TH. Since endogenous TH levels rise sharply during spontaneous metamorphosis, a similar sequence of events would be anticipated. However, after midclimax, CPS activity continues to increase, while plasma TH levels steadily decline, suggesting that other factors are involved. To obtain insight into this problem, the changes in CPS mRNA level during spontaneous development were determined using a mammalian CPS cDNA probe and correlated with changes in CPS activity and plasma T3 concentration. CPS mRNA level and CPS activity were barely detectable until midprometamorphosis, but both increased rapidly during the latter half of this phase. CPS activity continued to rise, reaching a maximum in the adult frog. The CPS mRNA level, however, was highest during the first half of climax, but declined after midclimax and was relatively low in the adult frog. Studies were also performed in which the rise and fall in the plasma T3 concentration typical of metamorphic climax were induced by exposure of premetamorphic tadpoles to T3, followed by its withdrawal. Both CPS activity and CPS mRNA level were induced by T3, but when plasma T3 levels fell after removal of the exogenous T3, CPS mRNA level, but not CPS activity, also decreased. Additional studies indicated that the TH-induced increase in CPS mRNA was evident within 24 h, could be prevented by simultaneous injection of actinomycin-D, and could not be induced in tadpoles undergoing climax; in this phase the T3 receptors are fully occupied with endogenous TH. When premetamorphic tadpoles were immersed in T3-containing water (0-500 nM) for 6 days, CPS mRNA, CPS activity, and plasma T3 concentration increased in parallel, reaching a maximum at 50 nM. At 50 nM T3, the plasma T3 level was sufficient to saturate the receptors, and no additional increase in CPS mRNA level or CPS activity was obtained at higher concentrations of T3. These studies indicate that the CPS mRNA level during spontaneous development correlates with the plasma T3 concentration and suggest that it is a function of T3 receptor occupancy. The data are also consistent with an effect of TH on transcription of the CPS gene and with a relatively long half-life of its protein product, the CPS enzyme.

Animals

Decrease in polylactosaminoglycans associated with lysosomal membrane glycoproteins during differentiation of CaCo-2 human colonic adenocarcinoma cells.

The proportion of labeled polylactosaminoglycans found in glycoproteins decreases during spontaneous differentiation of CaCo-2 human colonic adenocarcinoma cells to enterocytes in culture (A. Youakim and A. Herscovics, Biochem. J., 247: 299-306, 1987). To identify polylactosaminoglycan-containing glycoproteins, CaCo-2 cells were incubated with [3H]glucosamine or [3H]fucose, for 24 h, and membrane glycoproteins solubilized with 0.5% Nonidet P-40 were fractionated by affinity chromatography on Datura stramonium (DSA)-agarose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography showed that a restricted set of glycoproteins with a molecular weight of about 100,000 bound to DSA-agarose. These labeled glycoproteins were shown to contain polylactosaminoglycans by DSA-agarose chromatography and endo-beta-galactosidase digestion of Pronase-derived glycopeptides. Immunoprecipitation of the [3H]glucosamine-labeled Nonidet P-40 extract with polyclonal antibodies to the lysosomal membrane proteins h-lamp-1 and h-lamp-2 followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography also revealed a band with a molecular weight of about 100,000. The immunoprecipitates were digested with Pronase, and the resulting glycopeptides were first fractionated on Bio-Gel P-6 into excluded (Fraction I) and included (Fraction II) glycopeptides, and then by DSA-agarose affinity chromatography. A much greater proportion of labeled glycopeptides in undifferentiated cells (3 to 5 days in culture) than in differentiated cells (19 to 27 days in culture) was recovered in Fraction I; these glycopeptides were bound to DSA-agarose and were sensitive to endo-beta-galactosidase. This decrease in polylactosaminoglycans was associated primarily with h-lamp-1. These results indicate that h-lamp-1 of CaCo-2 cells contains polylactosaminoglycans and that it undergoes a change in glycosylation with differentiation.

Adenocarcinoma

Percutaneous absorption of triadimefon in the adult and young male and female rat.

The percutaneous absorption of 14C-phenoxy ring labeled triadimefon was studied in adult and young male and female Sprague-Dawley rats. Triadimefon was applied (41.1 to 46.4 micrograms/cm2) in 0.2 ml of acetone to areas comprising 3% of the body surface (7.0 to 14.5 cm2). Thirty-six animals were treated at the initiation of each study. Groups of three animals were subsequently killed at 1, 4, 8, 12, 24, 48, 72, 96, 120, 144, 168, and 192 hr after treatment. Skin from the treated area as well as blood, heart, liver, kidneys, remaining carcass, urine, and feces were analyzed for 14C by scintillation counting techniques. Based on 14C counts, triadimefon was lost more rapidly from the skin of young animals (t 1/2, 20 to 25 hr) than from the skin of adult animals (t 1/2, 29 to 53 hr). Recovery studies indicated that adult males, adult females, young males, and young females, respectively, absorbed 53, 82, 57, and 52% of the dose. The rest of the dose based on material balance was presumably lost by evaporation. Approximately 2.5 to 3.9% of the dose penetrated the skin in one hour and was available for absorption. The rate of entry triadimefon into blood was 2 to 2.5 times faster for young than that observed in adult animals. Elimination of it from blood was faster in the case of the young animals. Triadimefon was absorbed through the skins of the adult male, adult female, young male, and young female rats, respectively, at rates of 0.20, 0.50, 0.58, and 0.48 micrograms/hr/cm2 of skin.

Absorption

Percutaneous absorption and dermal dose-cholinesterase response studies with parathion and carbaryl in the rat.

Dermal dose-ChE response (toxicity) and percutaneous absorption of parathion and carbaryl were studied in the rat. Parathion was as toxic to young animals as it was to adults of the same sex and more toxic to adult females than to adult males. Carbaryl was nontoxic at administered dosages. In each percutaneous absorption study, [14C]parathion or [14C]carbaryl was applied to back skin (12.5-13.8 cm2) at 43.5 to 48 micrograms/cm2. Thirty-six rats were treated and three were killed at time intervals between 0.5 and 168 hr. The area of the skin which had been treated, plasma, heart, liver, kidneys, urine, feces, and the remaining carcass were analyzed for 14C. Recovery studies indicated that adult male and female rats, respectively, absorbed 59.2 and 57.0% of the applied parathion, while adult males absorbed 57.7% of the applied carbaryl. Parathion was lost from skin (t1/2, 28.6 hr) more rapidly than carbaryl (t1/2, 40.6 hr). Approximately 1.4 and 5.8%, respectively, of the applied parathion and carbaryl penetrated the skin within 1 hr and was available for absorption. Parathion was absorbed through skin of adult male and female rats, respectively, at rates of 0.33 and 0.49 micrograms hr-1 cm-2. Carbaryl was absorbed by male rats at the rate of 0.18 micrograms hr-1 cm-2. The half-lives for absorption of parathion by blood ranged between 0.38 and 2.1 hr, while elimination half-lives ranged between 28.6 and 39.5 hr. Carbaryl absorption and elimination half-lives were 1.26 and 67 hr, respectively.

Animals

Scanning electron microscopy of experimental Trichophyton mentagrophytes infections in guinea pig skin.

Trichophyton mentagrophytes invasion of guinea pig skin was examined by scanning electron microscopy. Biopsies were obtained daily for 12 days from experimental infection sites. Dermatophyte invasion, examined in detail by scanning electron microscopy of cross-sectioned, prefixed skin was evidenced by: the appearance of hyphae within the stratum corneum; follicular invasion by hyphae, which remained initially within the follicle wall; emergence of the hyphae from the wall into the follicular canal; proliferation of the fungus down the follicle, with furrowing of the follicle wall and hair shaft cuticle; penetration of hyphae into the hair shaft by subcuticular and transcuticular routes; and massive peripilar hyphal proliferation with arthrosporogenesis. A three-dimensional perception of the invasion sequence of a dermatophyte in guinea pig skin was obtained by scanning electron microscopy.

Animals

Development of arthrospores of Trichophyton mentagrophytes.

Arthrosporogenesis of the dermatophyte Trichophyton mentagrophytes was examined by light and by scanning and transmission electron microscopy. Sabouraud dextrose agar plates were inoculated with microconidia and incubated in an atmosphere of 8% CO2. Typical germination and hyphal branching continued to day 4, when hyphae began to be increasingly coated with granular-fibrillar material. Multiple replication of nuclei and formation of segregating septa followed. By day 6 the thick surface mesh sometimes was restricted to protruding rings, probably over septa. Between days 6 and 7, after thickening of outer and septal walls, units began to round and separate. Triangular gaps, which developed at the junction of septa and outer wall layers, enlarged so that spores were held together at their poles and along a tangential ring. With elongation of the spore to its barrel shape, the halves of the septum separated and the ring pulled apart, leaving a jagged, circular flange originating from the outer layer of cell wall extended toward the poles, covering the apparently exposed inner wall layer. Newly formed arthrospores, which measured 2.0 to 3.3 by 2.9 to 3.8 micronm and possessed walls of about 0.33-micronm thickness, has smooth sides but somewhat rough poles.

Spores, Fungal