Hydroxyproline (HYP) is a post-translational product of proline hydroxylation catalyzed by an enzyme prolyl 4-hydroxylase which plays a crucial role in the synthesis of all collagens, because the 4-hydroxyproline residues are essential for the folding of the newly synthesized collagen polypeptide chains into triple-heical molecules. Considering the role of collagen and its significance in many clinically important diseases such as liver cirrhosis, a great deal of attention has been directed toward the development of an assay at cell-based system. (omitted)
The objective of this study was to investigate the effect of cryopreservation by slow and rapid freezing on the sperm motility index, viability and morphology of post-thaw human spermatozoa. After rapid freezing and thawing, sperm motility index was significantly higher (MOT:47.40$\pm$20.06%, VCL : 38.12$\pm$15.58 $\mu$m/s, VSL : 28.19$\pm$14.10 $\mu$m/s, VAP:33.64$\pm$15.15 $\mu$m/s, and HYP 2.77$\pm$2.71%) than slow freezing and thawing(MOT : 43.39$\pm$ 18.79%, VCL .33.91 $\pm$ 13.50 Um/s, VSL . 19.98$\pm$0.88 $\mu$m/s, VAP : 24.60$\pm$11.72 $\mu$m/s, and HYP . 1.33$\pm$1.57% ; P<0.05). But sperm Linearity(LIN) was significantly lower(28.83 $\pm$ 10.35) comparing to the slow freezing method(34.64 $\pm$ 11.36 ; P<0.05). On the other hand, significant difference were not observed MAD, WOB, DNC and DNM by slow and rapid frozen-thawed methods. After rapid freezing and thawing, sperm viability was lower(60 $\pm$ 2.2%) than slow freezing method(62 $\pm$2.1%) and sperm morphology was higher(46$\pm$7.7%) than that(44: 8.3). But there was no significantly These results indicate that rapid freezing method was positive effect of sperm cryopreservation in human.
park Yeun Woo;Yang Si Yang;Lee Min Kyung;Jin Ju Young;Cho Jung Hee;Kim Ki Young
Journal of Physiology & Pathology in Korean Medicine
/
v.18
no.3
/
pp.868-873
/
2004
Renal dysfunction could be developed as the secondary disease of liver cirrhosis. Delayed or suppresed lipid peroxidation by the treatment with physiological active substances could be explained as the antioxidative and protective effect in tissue damage. In this study, we investigated an antioxidative effect and renal function improvement of Hovenia dulcis in liver fibrosis(cirrhosis) induced rats. The female Sprague-Dawley rats (180∼210 g) were divided into 3 groups (Normal, AC: CCl₄ mixture treated group, AC-HV: CCl₄ mixture+ Hovenia dulcis treated group) and renal damage was developed by CCl₄ mixture administration in 4 weeks (0.8 ㎖/rat). The tissue of kidney and liver and sera were used for quantitative measurement of enzyme activity, MDA and Hyp. The histological change and gene expression of collagen α1(III) mRNA and a1(IV) mRNA were observed by Masson's trichrome staining and RT-PCR. As a result, the clinical biochemical parameters of liver function (AST and ALT) in sera of AC-HV group showed significantly 46.4% and 104.8% lower (p<0.005), and the level of ALP and BUN as the parameter of protein urine and azotemia showed 17.8 % and 25.8 % lower than in AC group. In AC-HV group, the concentration of MDA in kidney and liver was decreased significantly 15.8% and 21.3% when compared with AC group (p<0.01 -0.005). The content of Hyp in kidney of AC-HV group is merely higher than in AC group, in contrast to liver tissue. The expression of collagen α1(III) mRNA and collagen α1(IV) mRNA was decreased in AC, but both of collagen mRNA in normal and AC-HV group expressed fast similar. More massive lipid droplets, thicker collagen fiber bundles in portal triads and more formation of portal central septum were observed in the liver of AC group than in AC-HV group. In conclusion, CCl₄ mixture intoxication could be developed not only liver fibrosis(cirrhosis) but also renal dysfunction by the massive lipid peroxidation and suppression of interstitial collagen and basement membrane collagen synthesis. And the water extract of Hovenia dulcis may be possessed the antioxidative and protective effect and improvement of kidney function in renal dysfunction induced rats.
Lee Min Kyung;Park Sung Hye;Seo Eui Suok;Kim Ki Young
Journal of Physiology & Pathology in Korean Medicine
/
v.18
no.5
/
pp.1485-1489
/
2004
Panax ginseng is the one of best famous phytochemical plant in the world and it's various positive effects such as antioxidant, regulation of immunity are very well known. In this study, we investigated primary the cell viability and morphological change and secondary an antioxidative effect and liver function improvement of extract from Ginseng folium and stem in CCl4 intoxicated rats. The NCTC cell line were used for cell viability and sirius red staining before the animal experiment. The female Sprague-Dawley rats (90-100g) were divided into 3 groups (Normal, AC: CCl₄ treated group, GFS: CCl₄+ extract of Ginseng folium and stem treated group) and acute liver damage was developed by one time administration of CCl₄ mixture (0.5㎖/rat). The liver tissue and sera were collected and used for quantitative measurement of enzyme activity (AST, ALT, ALP, BUN), MDA and Hyp. As a result, cell viability in GFS treated group (in concentration of 3.33-33.33㎎ GFS/200㎕ medium) was 180.9-241.0% significantly and dose dependently higher than in control group. And potential state of cell growth and differentiation and no criteria of cytoplasm lysis and nucleus breaking were observed in control and GFS group. The parameters of liver function (AST and ALP) in sera of GFS group showed significantly 93% and 67.6% lower than AC group (p<0.005-0.05). And the level of ALT and BUN showed fast similar in AC group and GFS group. The concentration of MDA in liver was decreased 576.5% significantly in GFS group when compared with AC group (p<0.005). The content of Hyp in GFS group is merely lower than in AC group. In conclusion, the water extract of Ginseng folium and stem such as Ginseng radix may be possessed the antioxidative effect and improvement of liver function in CCl₄ intoxicated rats.
PARK Choon-Kyu;KIM Woo-Jun;KANG Hoon-I;KANG Tae-Jung;SHIN Suk-Woo
Korean Journal of Fisheries and Aquatic Sciences
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v.27
no.2
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pp.140-148
/
1994
Indentification of IMP was carried out and changes in ATP breakdown products during storage at $0^{\circ}C\;ad\;20^{\circ}C$ were investigated in the muscles of ascidian Halocynthia roretzi. For identifying IMP, the ion-exchange column chromatographic method was applied to the perchloric acid extract of the muscle of cultured ascidian collected at the southern coast near Chungmu of Korea in April 1989. The IMP of sample was eluted a little earlier than that of the reference standard, but absorption spectra of both fractions agreed each other. In addition, both fractions gave the identical retention time of HPLC. These results reconfirmed that the ascidian muscle did contain IMP, indicating that ATP was degraded through IMP breakdown pathway, such as $ATP{\to}ADP{\to}AMP{\to}IMP{\to}Ino{\to}Hyp$. Ado was detected in some samples and IMP was detected throughout the experimental periods at both temperatures, but their levels were always very low; they did not increase significantly even when the decreasing rate of AMP was very rapid and concomitant remarkable increase in Ino were observed at the early stage of storage. Those changes in ATP suggest that AMP deaminase activity was present in the ascidian muscle, though it was very low. The main breakdwon pathway of ATP was assumed to be $ATP{\to}ADP{\to}AMP{\to}Ado{\to}Ino{\to}Hyp$. In conclusion, there were two breakdown pathways of ATP in the muscle of ascidian as was the case for the muscle of many marine crustaceans.
The chronic cholestasis induce to biliary liver fibrosis (cirrhosis) and the increased products of ROS(reactive oxygen species) cause to the liver damage. In this study ; the antioxidant and antifibrotic effect of dried extracts of oriental medicine (DW) was investigated under the liver fibrotic (cirrhotic) condition. The female Sprague-Dawley rats were divided in 5 groups (Normal, Op-2, Op-4, OpDW-2, OpDW-4). Except for normal group, the rats were induced to biliary liver fibrosis (cirrhosis) by the operation of bile duct ligation/scission (BDU/S) and were observed in 2 weeks or 4 weeks. And the prepared DW was treated p.o.2 ml/day/rats in 2 weeks or 4 weeks for OpDW groups. At the time of sacrifice, the liver, kidney, spleen were weighed and the ratio of organ weight/body weight was calculated. The MDA, the hyp and biochemical parameters (GOT GTP, ALP, t-bili) were measured in sera and liver tissue of rats. The biochemical change was observed on liver tissue. In the result, the hepatomegaly and spleenomegaly appeared in all BDL/S operated rats, and significantly lower liver weight was observed in OpDW-4 group compared with in Op-4 group (p<0.05). The level of clinical parameters in sera of all liver fibrosis (cirrhosis) developed rats was higher than in normal group. Especial1y, the value of GOT in OpDW-2 group and ALP in OpDW-4 group showed significantly lower than in Op-2 group and Op-4 group (p<0.01, p<0.005). The content of hyp in all operation groups was significantly higher than in normal group (p<0.05∼<0.005), and showed significantly lower value in the OpDW-4 group than in Op-4 group (p<0.05). The product of lipid peroxidationUDA) increased significantly under the fibrotic(cirrhotic) condition (p<0.05∼ <0.005), and the MDA value in OpDW-4 group decreased significantly in Op-4 group (p<0.005). The histological change (bile duct proliferation, fibrosis, collagen bundle) was similarly observed in Op-2 group and in OpDW-2 group, but the weak fibrosis and bile duct proliferation were observed in OpDW-4 group compared with in Op-4 group. In conclusion, lipid peroxidation and severe liver damage were activated by bile duct obstruction, and the measurement of MDA and hap can be useful monitor for the screening of antioxidant and antifibrotic effect in experimental liver fibrosis (cirrhosis). The 4 weeks treatment with DW extracts suppressed lipid peroxidation and inhibited fibrotic (cirrhotic) process in BDL/S operated rats.
Kim, Hong Seok;Hong, Won Kyu;Lee, Mun Hoe;Kim, Hyeong Min;Chung, Hee Chul;Lee, Jin Hee
Journal of the Society of Cosmetic Scientists of Korea
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v.47
no.2
/
pp.147-153
/
2021
Collagen hydrolysate (CH) is known to prevent skin aging by stimulating skin dermal fibroblasts to promote synthesis of extracellular matrix such as collagen and elastin. Recently, among the various factors that cause skin aging, advanced glycation end products (AGEs) have received particular attention. However, the effect of CH on AGE accumulation has not been studied. Since CH could affect AGE accumulation by promoting production of skin structural proteins, clinical trial was performed using low molecule collagen peptide (LMCP), which were CH containing 25% tripeptide and 4% Gly-Pro-Hyp. Skin autofluorescence (SAF) values were measured using an AGE reader to evaluate accumulation of AGE in skin. As a result of applying 0.5% and 1.0% LMCP solutions to the subject's forearm for 8 weeks, the SAF value at the test site significantly decreased compared to the control site. Additionally, in vitro test was performed using CCD-986sk to evaluate the promotion of collagen synthesis in skin fibroblasts by LMCP. As a result, 800 ㎍/mL of LMCP significantly increased synthesis of human pro-collagen Iα1 (COL1A1) in CCD-986sk. Through this study, we have confirmed that tropical LMCP applications can promote collagen synthesis to help anti-glycation effects, suggesting that LMCP has potential as an anti-aging cosmetic material.
Mutagenesis of protoplast could serve a great potential tool for improvement of strains and genetics in higher fungi. For the isolation of auxotrophic mutants from protoplasts of Pleurotus ostreatus and Pleurotus florida, viability levels of ultraviolet lights were determined. Seven auxotrophs were obtained from protoplasts irradiating UV to give $0.83{\sim}15%$ survival. The mutants showed a single requirement for each of Arg, Ribo-l, Ribo-2 or Phen for growth. Some of them showed two or three kinds of requirements, Gly Ser, Ade Hypo or Ala Om Tryp for growth.
This experiment was conducted to predict the effects of motional characteristics on the fertility of Korean native cattle(KNC) by using CASA technology and in vitro fertilization system. Twenty-six KNC frozen semen straws were obtained from Korean KNC improvement department, livestock improvement main division, national livestock cooperatives federation in Korea. Specimens were allowed to thaw at $37^{\circ}C$ for 30 sec in water bath. Semen analysis was performed on semen image analysis system(SIAS, Medical supply, Korea) adjusted to the gate settings and used the semen droplet ($5{\mu}l$) placed on Makler counting chamber(Sefi medical instrument, Israel) prewarmed at $37^{\circ}C$. The same person used the same micropipette to fill the Makler counting chamber. A total of 150 or more of sperms were analysed in each specimen by a single trained person by scanning at least 5 to 10 fields. The oocytes collection, in vitro maturation, IVF, in vitro culture and determination of the cleavage rate were performed by the technique, as described by Hwang et al (1997). Statistical analysis was done by linear regression with use of the Sigma plot program on a IBM personal computer. The cleavage rate in vitro fertilized oocyte was significantly correlated(P<0.05) with MOT, VCL, VSL, VAP, ALH, BCF and MAD, but not CON, LIN, STR, WOB, DNM, DNC and HYP in regressional analysis. The results show that some kinematic characteristics of frozen-thawed semen by CASA can be predict the fertility in in vitro model system.
This study investigated the effects of protease treatments (trypsin, chymotrypsin, and pepsin) under various pressure levels (0.1-300 MPa) for the characteristics of porcine placenta hydrolysates. According to gel electrophoretic patterns, the trypsin showed the best placental hydrolyzing activity followed by chymotrypsin, regardless of the pressure levels. In particular, the peptide bands of tryptic-digested hydrolysate were not shown regardless of applied pressure levels. The peptide bands of hydrolysate treated chymotrypsin showed gradual decreases in molecular weights ($M_w$) with increasing pressure levels. However, the pepsin did not show any evidences of placental hydrolysis even though the pressure levels were increased to 300 MPa. The gel permeation chromatography (GPC) profiles showed that the trypsin and pepsin had better placental hydrolyzing activities under high pressure (particularly at 200 MPa), with lower $M_w$ distributions of the hydrolysates. Pepsin also tend to lower the $M_w$ of peptides, while the major bands of hydrolysates being treated at 300 MPa were observed at more than 7,000 Da. There were some differences in amino acid compositions of the hydrolysates, nevertheless, the peptides were mainly composed of glycine (Gly), alanine (Ala), hydroxyproline (Hyp) and proline (Pro). Consequently, the results indicate that high pressure could enhance the placental hydrolyzing activities of the selected proteases and the optimum pressure levels at which the maximum protease activity is around 200 MPa.
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