• Toxicological Research
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• v.5 no.2
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• pp.89-96
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• 1989

Rapid and complete epidermal-dermal separation procedure were determined in neonatal rat skin by light microscopic observation and by compairing enzyme activities in the separated epidermis. Microscopic appearance demonstrated the at four different separation procedures used in the study resulted in good separation of epidermis from dermis` heating method (i.e., immersion in 55C water for 30 sec, followed by immersion in 0-4C water) and microwave irradiation for 10 sec were saving time.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.601-606
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• 2001

A simple assay method of recombinant human epidermal growth factor (rhEGF) in a pharmaceutical preparation was studied and validated by capillary electrophoresis (CE) using micellar electrokinetic chromatography (MEKC) techniques. Factors affecting the migration behavior and separation performances of the peptide; type of buffers pH, butler concentration, and concentration of sodium dodecyl sulfates (SDS) were investigated to optimize the analytical performance. CE was performed using running buffers 50.0 mM borate (pH 8.5) containing 12.5 mM SDS at 20 $mutextrm{V}$ of the applied voltage. Calibration curves for the rhEGF showed good linearity (r>0.999) over the wide dynamic range from 1.25 to $100{\mu\textrm{g}}/ml$. Sample analysis was performed by using standard addition method to eliminate the matrix effects of dosage vehicle. This method is assumed to be useful for quality control (QC) of various forms of pharmaceutical products of the peptide.

• Journal of the Korean Society of Clothing and Textiles
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• v.17 no.3
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• pp.491-505
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• 1993

The influences of protease on the removal of various protein soils from cotton fabrics were studied. The human epidermal stratum corneum, hemoglobin and casein were used as protein soils. The soiled fabrics were denatured by steaming for 30 min. before washing and laundered using Terg-O-Tometer under washing conditions. The removal efficiency was evaluated by analysis of protein on the fabrics before and after washing by means of copper-Folin method. The relations between the removal and the characteristics of protease were discussed. Also the degradation of protein were examined by microscopy. The seperation of human epidermal stratum corneum after hydrolysis was examined by SDS-PAGE. The results obtained were as follow : 1. The protein from the soiled cotton fabric was removed effectively by adding protease. The removal of protein was increased in proportion to increasing of the enzyme concentration up to a certain point, but it began to decrease above the point. The removal effect was high in the order of casein>human epidermal stratum corneum>hemoglobin. Especially the protein was more effectively removed in ADS solution(pH 9.5) containing enzyme. 2. When protease was used with ADS. the removal of protein was efficiently showed in relatively short time(5~15min.) compared to using ADS only. It is due to the properties of this enzyme that reacts with very short time. 3. Even at low temperature the removal efficiency of enzyme was relatively higher compared with the activity of enzyme. The removal of protein soil was increased up to a maximum near $50^{\circ}C$, and then decreased. 4. The removal of protein by protease was improved with the increase of alkalinity in the pH range from 9.5 to 11.0 but it began to decrease above pH 11.0. 5. According to the increase of mechanical agitation, the removal effect was increased. But the removal efficiency of protease was more effective compared with the agitation in detergency. 6. According to the SDS-PAGE separation and micrograph it was confirmed that the human epidermal corneum was effectively hydrolysed by the enzyme added. So the fragments of protein were removed more efficiently by means of the interfacial reaction of AOS.

• KSBB Journal
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• v.8 no.3
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• pp.287-294
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• 1993

For the purpose of combining the purification processes for several biologically active proteins form human urine, an efficient integrated fractionation procedure has been investigated. The procedure was started by concentration with ultrafiltration and pH precipitation followed by a selectable combination of chromatography on gel filtration, adsorption, ion exchanger, affinity, and reverse phase column. By this process, the purified urokinase, epidermal growth factor and albumin migrated as a single band on SDS-polyacrylamide gel electrophoresis and were fully active. The recoveries of these purified proteins were 48%, 17%, and 46%, respectively.

• Korean Journal of Veterinary Service
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• v.34 no.1
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• pp.55-61
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• 2011

Dermatopathologic findings on cutaneous streptothricosis (bovine dermatophilosis) were conducted in Dinajpur of Bangladesh for one year. The animals were examined clinically at hospital during the physical visit of the affected animals and clinical manifestations and patients' history were recorded. The gross lesions were systematically examined, noted and categorized. An impression smear was made by the skin scraps, stained with Gram's stain and examined microscopically for the presence of causative bacteria. The suitable sizes of skins of 3 animals typically affected with dermatophilosis were collected surgically from the animals, and prepared for the histopathological study. The cardinal clinical features and gross lesions were characterized as paint-brush cutaneous surface with matted hairs, scab and crust formation, roughened coat; istopathologically as hyperkeratosis, parakeratosis, typical "palisading encrustations", extensive proliferation of coccoid shaped, bluish coloured bacteria (zoospores) in the cells of the outer rootsheath of the hair follicles, separation of keratinized layer from the epidermal epithelium, epidermal hyperplasia, densely cellular dermis with moderate destruction of glandular structures. Coccoid shaped gram positive organisms which form multidimensional arrangement were also observed by impression smear. The clinical signs, gross lesions, morphology of the organisms, staining properties, and histopathological features of the affected skin were diagnostic criteria for the bovine cutaneous streptothricosis.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.12
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• pp.1822-1829
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• 2003

Porcine colostrum and milk were separated into the acid-soluble and casein fractions by acidification followed by centrifuge. The acid-soluble fraction of porcine colostrum was further separated by liquid chromatography and anisotropic membrane filtration. Trypsin and chymotrypsin inhibitory capacity in porcine colostrum, milk and their components was determined by incubating bovine trypsin or chymotrypsin in a medium containing their corresponding substrates with or without addition of various amounts of porcine colostrum, porcine milk or their components. The inhibition of insulin-like growth factor I (IGF-I) and epidermal growth factor (EGF) degradation in pig small intestinal contents by porcine colostrum was measured by incubating iodinated IGF-I or EGF with the intestinal contents with or without addition of porcine colostrum. Degradation of labeled IGF-I or EGF was determined by monitoring the generation of radioactivity soluble in 30% trichloroacetic acid (TCA). The results showed that porcine colostrum had high levels of trypsin and chymotrypsin inhibitory activity and increased the stability of IGF-I and EGF in pig intestinal contents. The inhibitory activity declined rapidly during lactation. It was also found that trypsin and chymotrypsin inhibitory activity and the inhibition on IGF-I and EGF degradation in the acid-soluble fraction were higher than that in the casein fraction. Heat-resistance study indicated that trypsin inhibitors in porcine colostrum survived heat treatments of $100^{\circ}C$ water bath for up to 10 min, but exposure to boiling water bath for 30 min significantly decreased the inhibitory activity. Compared with the trypsin inhibitors, the chymotrypsin inhibitors were more heatsensitive. Separation of the acid-soluble fraction of porcine colostrum by liquid chromatography and anisotropic membrane filtration revealed that the trypsin and chymotrypsin inhibitory capacity was mainly due to a group of small proteins with molecular weight of 10,000-50,000. In conclusion, the present study confirmed the existence of high levels of protease inhibitors in porcine colostrum, and the inhibition of porcine colostrum on degradation of milk-borne growth factors in the pig small intestinal tract was demonstrated for the first time.

• Journal of Embryo Transfer
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• v.15 no.1
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• pp.67-75
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• 2000

This study was conducted to establish the optimal conditions for in vitro embryo production using oocytes derived from follicles of slaughter-house ovaries. The ovaries of Hanwoo were obtained from a local slaughter-house. The oocytes were aspirated from visible follicles of 2~7mm in diameter. The recovered oocytes which were completely surrounded by at least 2 layers of cumulus cells and a homogeneous cytoplasmic pigmentation were used. The selected oocytes were washed 3 or 4 times with D-PBS containing 10% bovine calf serum (BCS) and matured in vitor (IVM) in Ham's F-10 supplemented with 10% BCS or 0.01 $\mu\textrm{g}$/ml epidermal growth factor(EGF) at 39$^{\circ}C$ under 5% CO2 in air for 24 hours. They were fertilizqed in vitro (IVF) with fresh sperm separated by Percoll density gradient or swim-up in TALP media. The zygotes were cultrued with or without bovine oviductal epitherial cells(BOEC) in media(HECM-6 supplemented with 11 amino acid and / or TCM-199 supplemented with 10% BCS) for 7 to 10 days. The results obtained were as follow: The cleavage rate and developmental rate to blastocyst after maturation and IVF were not significantly different between Ham's F-10 with EGF(76.0% vs. 44.0%) and BCS(75.9% vs. 43.6%)(P<0.05). The cleavage rate and development rate to blastocyst after fertilizing by swim-up or Percoll method were not signifciantly(P<0.05) different between swim-up (80.2% vs. 29.2%) and Percoll(81.9% vs. 26.5%) (P<0.05). The cleavage rate in TCM 199(80.5) was signficiantly higher than that in HECM-6 (72.0%) (P<0.05). However, developmental rate to blastocyst using TCM 199 following HECM-6 for 3 or 4 days (42.2%) was significantly higher than that in TCM-199 alone(26.7%)(P<0.05). The cleavage rate and development rate of embryos produced in vitro by exchange timing for HECM-6 media were not significantly different between in day 3(78.6% vs. 45.5%) and day 4(75.0% vs. 43.2%)(P<0.05). The cleavage rate and developmental rate to blastocysts according to co-culture system were not significantly different between with (74.2% vs. 41.4%) and without BOEC(73.95 vs. 43.5%) (P<0.05). The number of blastomere in blastocyst stage after co-culture with or without BOEC was not significantly different (106.7$\pm$5.1 and 96.6$\pm$4.0). In conclusion, the most transferable IVP embryos could be produced from Ham's F-10 medium for IVM, Percoll density gradient method for IVF sperm separation and in vitro culture in HECM-6 until day 3 or day 4, and then transferred into TCM-199 until day 9 within adequate embryo density in culture droplets after insemination.