The glycation of BSA leads to protein/peptide modifications that result in the formation of AGEs. AGEs react with the amino groups of N-terminal amino acid residues, particularly arginine and lysine residues. Enhanced AGE formation exists in the blood and tissues of diabetics, as well as in aging and other disorders. The Identification of AGEs is of great importance. Mass spectrometry has been applied to identify and structurally elucidate AGEs. Here, we report on the identification of AGE-peptides and AGE precursors based on relative mass changes as a result of specific AGE formation. HPLC-ESIMS, ESI-MS/MS, and the Mascot database were used. The relative mass changes due to the specific type of AGE formation were added to the identified peptides followed by a manual search of the glycated samples, which resulted in the identification of seven peptides for the formation of five AGEs, namely CML, pyrraline, imidazolone A, imidazolone B, and AFGP. Four glycated peptides (FPK, ECCDKPLLEK, IETMR, and HLVDEPQNLIK) were identified in the formation of AGE-precursors.
Proceedings of the Korean Society of Applied Pharmacology
/
1995.04a
/
pp.71-71
/
1995
The succinic semialdehyde dehydrogenase which is one of the key enzyme of GABA shunt in CNS has been purified from bovine brain homogeneously for the first time. The molecular mass of the native enzyme was estimated to be approximately 110,000 on gel filtration, The subunit molecular mass was determined by SDS-PAGE to be 54,000. These results indicate that the enzyme is a dimeric protein made up to identical subunits. Chemical modification studies of the enzyme suggest that the critical lysyl, connected with catalytic activity of the enzyme, The binding of IAF-SSDH(enzyme tagged with fluoreceine) to GABA transaminase which catalyzes the degradation of GABA was monitored by steady emission anisotropy. The changes of fluorescence anisotropy by interactions between two enzymes suggest that the formation of enzyme cluster must be invoved in the regulation of GABA concentration in brain tissues. The inhibitory effects of some antiepileptic and anticonvulsant drugs on the enzyme were also examined.
Park, Samina;Kim, Soo Hwan;Lim, Hong-Gook;Lim, Cheong;Kim, Yong Jin
Journal of Chest Surgery
/
v.46
no.1
/
pp.1-13
/
2013
Background: Glutaraldehyde (GA) is a widely used cross-linking agent for improving mechanical properties and resistance to enzymatic degradation of collagenous tissue, but it has several drawbacks such as calcification and cytotoxicity. The aim of this study was to find the alternative effective cross-linking methods to GA. Materials and Methods: Bovine pericardium was processed with GA with ethanol+octanol and glycine detoxification, and polyethylene glycol (PG) space filler, dimethyl 3,3'-dithiobispropionimidate (DTBP), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) treatment, and the physical fixation of ultraviolet irradiation were done. The biologic material properties of variously treated pericardial tissues were assessed by biochemical, mechanical and histological tests. Treated pericardial tissues were also implanted subcutaneously or intramuscularly into the rabbit for 10 weeks to assess the xenoreactive antibody response of immunoglobulin G and M, their anti-calcification effect. Results: The biochemical and mechanical properties of EDC fixed pericardial tissues were comparable to the GA fixed tissue. The cytotoxicity was lowest in space filler treated GA fixed group. In rabbit subcutaneous or intramuscular implantation models, decellularization, space filler, EDC treatment group showed significantly lower calcium content than GA only and DTBP treatment group (p<0.05, analysis of variance). The titer of anti $Gal{\alpha}1-3Gal{\beta}1$-4GlcNAc-R antibodies did not change in the postimplantation serial enzyme-linked immunosorbent assay. Hematoxylin and eosin and von Kossa staining showed that decellularization, space filler, EDC, and ultraviolet treatment had less inflammatory cell infiltration and calcium deposits. Conclusion: The decellularization process, PG filler, and EDC treatments are good alternative cross-linking methods compared to GA only fixation and primary amine of DTBP treatment for cardiovascular xenograft preservation in terms of the collagen cross-linking stability and in vivo anti-calcification effects.
Telomeres consist of TTAGGG tandem repeated DNA sequences with specific proteins and locate at chromosome ends. Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. Telomerase is a ribonucleoprotein which has a template for the synthesis of telomeric DNA. This study was carried out to analyze the amount of telomeric DNA and telomerase activity in cattle cells. Analysis of the quantity of telomere in lymphocytes was done at different ages, sex and among Korean cattle and Holstein breeds. The telomerase activity was also analyzed in liver, brain, heart, kidney, and testis tissues of fetal calf and of 18 month old cattle. The amount of telomeres in lymphocytes and other tissue cells was analyzed by Quantitative-Fluorescence in situ Hybridization (Q-FISH) technique using a telomeric DNA probe. Telomerase activity was analyzed by Telomeric Repeat Amplification Protocol assay (TRAP). The amount of telomeric DNA on the lymphocytes during the whole life span was decreased along with age. Quantity of telomeres in Korean cattle was significantly higher than that in Holstein breed. The amount of telomeric DNA in males was significantly higher than that in females. Telomerase activity was up-regulated in most bovine tissues during fetal stage, but was down-regulated in most tissues at mature 18 month age except the testis cells. This study indicates that the amount of telomeres and telomerase activity of cells can be used as an age marker or/and a physiological marker of cattle.
Kim, SuJi;Lee, AhReum;Kim, SooHyun;Kim, KyeongJo;Kwon, OJun;Choi, JoonYoung;Koo, JinSuk;Roh, Seong-Soo
The Korea Journal of Herbology
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v.32
no.4
/
pp.53-60
/
2017
Objectives : Collagen decrease of Skin appears through various path ways. One of causes may be the Advanced glycation endproducts (AGEs) that combine formation of glucose and protein. The aim of this study was to explore the prevent wrinkle formation of Paeoniae radix (PR) and heated Paeoniae radix (HPR) via AGEs path way. Methods : AGEs formation inhibitory activities of PR and HPR measured using bovine serum albumin, glucose, and fructose. To evaluate the protective effects of PR and HPR in diabetic rats induced with streptozotocin (STZ) and methyl glyoxal (MGO), SD rat were distributed into four groups. Normal rats (Nor), AGEs-induced rats (Con), AGEs-induced rats treated with 100 mg/kg PR(PR), AGEs-induced rats treated with 100 mg/kg HPR (HPR). To induce AGEs, streptozotocin (50 mg/kg) was administered intraperitoneally and after 3 days administrated 100mM methyl glyoxal for 3 weeks. Results : The oral administration of HPR inhibited AGEs in skin tissues compared with PR. The increased reactive oxygen species (ROS) levels in the serum were diminished by HPR treatment. The analyses of kidney and skin tissues proteins indicated that HPR treatment effectively reduced AGEs related protein levels as compared to that by PR treatment. Also, HPR decreased anti-oxidant related protein levels in skin tissues such as catalase, glutathione peroxidase. Moreover, it inhibited the reduction of COL1A2 by decreasing MMP-1. Conclusion : Based on these results, it was suggested that PR and HPR could have Improving effects on wrinkle formation. These evidences provide useful information for the development wrinkle formation treated agent.
Journal of Dental Rehabilitation and Applied Science
/
v.23
no.3
/
pp.249-257
/
2007
State of problem : The use of zirconium oxide all-ceramic material provides several advantages, including a high flexural strength(>1000MPa) and desirable optical properties, such as shading adaptation to the basic shades and a reduction in the layer thickness. Along with the strength of the materials, the cementation technique is also important to the clinical success of a restoration. Nevertheless, little information is available on the effect of different surface treatments on the bonding of zirconium high-crystalline ceramics and resin luting agents. Purpose : The aim of this study was to test the effects of surface treatments of zirconium on shear bond strengths between bovine teeth and a zirconia ceramic and evaluate differences among cements Material and methods : 54 sound bovine teeth extracted within a 1 months, were used. They were frozen in distilled water. These were rinsed by tap water to confirm that no granulation tissues have left. These were kept refrigerated at $4^{\circ}C$ until tested. Each tooth was placed horizontally at a plastic cylinder (diameter 20mm), and embedded in epoxy resin. Teeth were sectioned with diamond burs to expose dentin and grinded with #600 silicon carbide paper. To make sure there was no enamel left, each was observed under an optical microscope. 54 prefabricated zirconium oxide ceramic copings(Lava, 3M ESPE, USA) were assigned into 3 groups ; control, airborne-abraded with $110{\mu}m$$Al_2O_3$ and scratched with diamond burs at 4 directions. They were cemented with a seating force of 10 ㎏ per tooth, using resin luting cement(Panavia $F^{(R)}$), resin cement(Superbond $C&B^{(R)}$), and resin modified GI cement(Rely X $Luting^{(R)}$). Those were thermocycled at $5^{\circ}C$ and $55^{\circ}C$ for 5000 cycles with a 30 second dwell time, and then shear bond strength was determined in a universal test machine(Model 4200, Instron Co., Canton, USA). The crosshead speed was 1 mm/min. The result was analyzed with one-way analysis of variance(ANOVA) and the Tukey test at a significance level of P<0.05. Results : Superbond $C&B^{(R)}$ at scratching with diamond burs showed the highest shear bond strength than others (p<.05). For Panavia $F^{(R)}$, groups of scratching and sandblasting showed significantly higher shear bond strength than control group(p<.05). For Rely X $Luting^{(R)}$, only between scratching & control group, significantly different shear bond strength was observed(p<.05). Conclusion : Within the limitation of this study, Superbond $C&B^{(R)}$ showed clinically acceptable shear bond between bovine teeth & zirconia ceramics regardless of surface treatments. For the surface treatment, scratching increased shear bond strength. Increase of shear bond strength by sandblasting with $110{\mu}m$$Al_2O_3$ was not statistically different.
The Stearoyl-CoA Desaturase(SCD) is a key enzyme, which converting palmitic acid(16:0) and stearic acid (18:0) to pahnitoleic acid(16:1) and oleic acid(l8:1), respectively. The concentration of oleic acid(18:1) in meat of beef cattle could influence both palatability and perception of meat. This experiment has conducted to determine relationship between the compositions of monounsaturated fatty acids and the SCD mRNA level in bovine liver and loin muscle tissue. The compositions of palmitoleic acid(16:1) and oleic acid(18:1) in loin muscle were 5% and 46% of total lipid and in liver were 2% and 20% of total lipid, respectively. On the other hand, the compositions of palmitic acid(16:0) and stearic acid(18:0) in loin muscle were 25% and 45% of total lipid and in liver were 14% and 43% of total lipid, respectively. The ratio of monounsaturated to saturated fatty acids(the desaturation index) was used as a measure of SCD activity in tissues. The average desaturation index in loin muscle was higher about 3.6-fold than that in liver. The desaturation index of oleate/stearate and palmitoleatelpalmitate in loin muscle were higher 8-fold and 1.8-fold than those in liver, respectively, showing that the substrate specificity of SCD enzyme was very different between liver and muscle tissues. To determine whether the composition of monounsaturated fatty acids in liver and muscle are dependent on SCD expression, SCD mRNA level was examined by RT-PCR analysis. The SCD mRNA level in loin muscle was higher about 3-fold than that in liver. Thus, the quantitative relationship between the desaturation index of fatty acid and SCD mRNA was observed in liver and muscle. The difference in the compositions of monounsaturated fatty acids between bovine liver and muscle tissues may be due to different level of Stearoyl-CoA Desaturase mRNA.
Purpose: The aim of this study was to elucidate the effect of a bovine hydroxyapatite/collagen (BHC) block in one-wall intrabony periodontal defects in dogs. Methods: A one-wall intrabony periodontal defect (4 mm wide and 5 mm deep) was prepared bilaterally at the mesial side of the mandibular fourth premolar in five beagle dogs. After thorough root planing, block-type BHC ($4{\times}5{\times}5$ mm) was placed on one side. The contralateral defect area did not receive any material as a sham-surgery control. Histological analysis of the sites was performed after an 8-week healing period. Results: Two of five samples in the experimental group healed well without dissipation of the graft materials, and histological analysis revealed excellent regeneration of the periodontal tissues. However, most of the grafted materials had been displaced in the other three samples, leaving only a small portion of the graft. The measured parameters exhibited large standard deviations, and the mean values did not differ significantly between the experimental and sham-surgery control sides. Conclusions: The application of BHC alone-without a barrier membrane-to wide, one-wall intrabony periodontal defects yielded inconsistent results regarding both periodontal regeneration and substantivity of the graft materials. Thus, the use of a barrier membrane for noncontained-type defects is recommended to improve the stability of the grafted material, and to condense it.
Kim, Young-Chool;Lim, Sung-Bin;Chung, Chin-Hyung;Lee, Chong-Heon
Journal of Periodontal and Implant Science
/
v.33
no.4
/
pp.693-703
/
2003
The present study evaluated the effects of guided tissue regeneration using xenograft material(deproteinated bovine bone powder), with and without Calcium sulfate membrane in beagle dogs. Contralateral fenestration defects (6 ${\times}$ 4 mm) were created 4 mm apical to the buccal alveolar crest of maxillary premolar teeth in 5 beagle dogs. Deproteinated bovine bone powders were implanted into fenestration defect and one randomly covered Calcium sulfate membrane (experimental group). Calcium sulfate membrane was used to provide GTR. Tissue blocks including defects with soft tissues which were harvested following four & eight weeks healing interval, prepared for histo-phathologic analysis. The results of this study were as follows, 1. In control group, at 4 weeks after surgery, new bony trabecular contacted with interstitial tissue and osteocytes lie cell were arranged in new bony trabecule. Bony lamellation was not observed. 2. In control group , at 8 weeks after surgery, scar-like interstitial tissue was filled defect and bony trabecule form lamellation. New bony trabecular was contacted with interstitial tissue but defect was not filled yet. 3. In experimental group, at 4 weeks after surgery, new bony trabecular partially recovered around damaged bone. But new bony trabecule was observed as irregularity and lower density. 4. In experimental group, at 8 weeks after surgery, lamella bone trabecular developed around bone cavity and damaged tissue was replaced with dense interstitial tissue. In conclusion, new bone formation regenerated more in experimental than control groups and there was seen observe more regular bony trabecular in experimental than control groups at 4 weeks after surgery. In control group, at 8 weeks after surgery, the defects was filled with scar-like interstitial tissue but, in experimental group, the defects was connected with new bone. Therefore xenograft material had osteoconduction but could not fill the defects. We thought that the effective regeneration of periodontal tissue, could be achieved using GTR with Calcium sulfate membrane.
The corpus luteum (CL) is formed by the action of a surge of luteinizing hormone (LH) on the pre-ovulatory follicle. Luteal cells derived from granulosa and theca interna cells continue to secrete progesterone for about two weeks. LH in domestic animals is essential for the normal secretion of progesterone at all stages of the luteal phase. For this process in the rodents, 20$\alpha$-hydroxysteroid dehydrogenase (20$\alpha$-HSD) is indispensable. 20$\alpha$-HSD is an enzyme to be a biologically inactive steroid. This enzyme plays a critical role in the regulation of the rat luteal function and reported to be present in steroid-producing tissues such as the testis and adrenal gland. We have purified 20$\alpha$-HSD and found two distinct 20$\alpha$-HSD molecules (HSD-1 and HSD-2). Their molecular weights are both estimated to be 33kd.The amino acid compositions of HSD-1 and HSD-2 are mostly similar, but there is a slight difference in the content of lysine. We demonstrated that 1) CL of previous generations contribute more to whole ovarian 20$\alpha$-HSD activity, 2) newly formed corpora lutea contain only 20$\alpha$-HSD-1 activity, and 3) old CL express activities of each HSD isozyme as shown in the luteal tissue of cycling rats on the day of diestrus where only degenerating old CL exist. The increase in 20$\alpha$-HSD activity identified seems to be related to the increase in the numbers of 20$\alpha$-HSD-positive cells. Interestingly, 20$\alpha$-HSD-1 activities were strongly found in the follicle fluids and theca interna cells by immunohistochemical study. Thus, the activity of 20$\alpha$-HSD may be related to a survival mechanism of those luteal cells and follicles remaining in the ovaries. Luteal cells arise from two sources. The small luteal cells are all of theca cell origin, while the large luteal cells are mainly of granulosa cell origin. CL of Korean Native Cattle, as those of other animal species, contains two morphologycally and functionally distinct luteal cell populations, such as small and large luteal cells as well as nonluteal cells. In all reproductive states except in the late luteal phase, the bovine CL also contained more small luteal cells than large luteal cells. Luteal tissue secretes a variety of growth factors (proteins) and the pattern of secretion changes during all stages of the luteal phase. These growth factors could be important in regulating the function of the bovine corpus luteum and may act in a potential endocrine autocrine and paracrine mechanisms. Therefore, further work has to be done to elucidate the role of growth factors in the ovary, especially in the corpus luterum. Interest should be focussed on interaction of these growth factors in the regulation of luteal cell and the localization of cytokine synthesis in differnet luteal cells.
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