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Investigation into the Distribution of Total, Free, Peptide-bound, Protein-bound, Soluble-and Insoluble-Collagen Hydroxyproline in Various Bovine Tissues

  • Siddiqi, Nikhat J.;Alhomida, Abdullah S.
    • BMB Reports
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    • v.36 no.2
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    • pp.154-158
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    • 2003
  • Collagen is a family of proteins which consists of several genetically distinct molecular species and is intimately involved in tissue organization, function, differentiation and development. The purpose of this study was to investigate the concentration of different hydroxyproline (Hyp) fractions viz., total, free, peptide-bound, protein-bound, soluble- and insoluble-collagen hydroxyproline (Hyp) in various bovine tissues. Results showed that liver had the highest concentration of free Hyp followed by kidney, brain, spleen, lungs, muscle and heart. Liver also had the highest concentration of peptide-bound collagen Hyp followed by kidney, heart, spleen, lungs, brain and muscle. The concentration of protein-bound collagen Hyp was highest in the liver, followed by kidney, spleen, lungs, muscle, brain and heart. Total Hyp was highest in the liver, followed by kidney, spleen, brain, heart, muscle and lungs. Liver also had significantly high concentration of collagen as compared to other tissues examined (P<0.001). Spleen had the significantly higher concentration of soluble-collagen Hyp when compared to other tissues (P<0.001). This was followed by heart, muscle, lungs, brain, kidney and liver. Heart had the highest concentration of insoluble-collagen Hyp followed by lungs, kidney, liver, muscle, spleen and brain. The variation among the insoluble-collagen Hyp concentration of heart and muscle, spleen and brain was significant (P<0.001). We speculate that these differences could be due to the variation in turn over of rate of collagen metabolism in this species.

Purification and Characterization of Antioxidative Peptides from Bovine Skin

  • Kim, Se-Kwon;Kim, Yong-Tae;Byun, Hee-Guk;Park, Pyo-Jam;Ito, Hisashi
    • BMB Reports
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    • v.34 no.3
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    • pp.219-224
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    • 2001
  • To identify the antioxidative peptides in the gelatin hydrolysate of bovine skin, the gelatin was hydrolyzed with serial digestions in the order of Alcalase, pronase E, and collagenase using a three-step recycling membrane reactor. The second enzymatic hydrolysate (hydrolyzed with pronase E) was composed of peptides ranging from 1.5 to 4.5 kDa, and showed the highest antioxidative activity, as determined by the thiobarbituric acid method. Three different peptides were purified from the second hydrolysate using consecutive chromatographic methods. This included gel filtration on a Sephadex G-25 column, ion-exchange chromatography on a SP-Sephadex C-25 column, and high-performance liquid chromatography on an octadecylsilane chloride column. The isolated peptides were composed of 9 or 10 amino acid residues. They are: Gly-Glu-Hyp-Gly-Pro-Hyp-Gly-Ala-Hyp (PI), Gly-ProHyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (PII), and Gly-ProHyp-Gly-Pro-Hyp-Gly-Pro-Hyp (PIII), as characterized by Edman degradation and fast-atom bombardment mass spectrometry. The antioxidative activities of the purified peptides were measured using the thiobarbituric acid method, and the cell viability with a methylthiazol tetrazolium assay The results showed that PII had potent antioxidative activity on peroxidation of linoleic acid. Moreover, the cell viability of cultured liver cells was significantly enhanced by the addition of the peptide. These results suggest that the purified peptide, PII, from the gelatin hydrolysate of bovine skin is a natural antioxidant, which has potent antioxidative activity.

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Determination of Hydroxyproline Contents of Pork Meat for Quality Evaluation of Meat Products (돈육 등급별 Hydroxyproline 함량 분석법을 이용한 육가공품의 품질 평가)

  • Kim, Jin-Seong;Kim, Jae-Hun;Park, Jin-Gyu;Park, Jae-Nam;Lee, Ju-Woon;Byun, Yu-Ryang;Byun, Myung-Woo
    • Food Science of Animal Resources
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    • v.26 no.3
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    • pp.362-367
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    • 2006
  • This study was conducted to evaluate the feasibility of determination of hydroxyproline (Hyp) contents in pork meat for the quality evaluation of meat products. The relationship between the Hyp contents and pork grade was not high in porcine semitendinosus muscle. Whereas, Hyp contents of pork fresh ham and hamburger patties prepared with them were significantly increased as the decrease of pork grade. These results showed that the determination of Hyp contents might be a useful method for the quality evaluation of meat products.

Determination of Free 4-hydroxyproline with Dansylchloride by HPLC in Human Urine (소변 중 4-hydroxyproline 분석에 관한 연구)

  • Lee, Keou-Weon;Cho, Young-Bong;Lee, Kyung-Jong
    • Journal of Preventive Medicine and Public Health
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    • v.35 no.4
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    • pp.282-286
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    • 2002
  • Objectives : The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. Methods : The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined.4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the a-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 mmol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). Results : The detection limit for the urinary free 4-Hyp was $0.364{\mu}mol/l$. The recovery rate of 4-Hyp was 99.7%, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and $0.32-56.27{\mu}mol/l$. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers ($11.82{\pm}6.73\;nmmol/mg\;Cre$) were 2-fold higher than in pre-shift workers ($5.36{\pm}5.53\;nmol;/mg\;Cre$) and controls ($4.91{\pm}4.89\;nmol;/mg\;Cre$). Conclusions : This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.

The Order of Normal Form Generalized Hypersubstitutions of Type τ = (2)

  • Sudsanit, Sivaree;Leeratanavalee, Sorasak
    • Kyungpook Mathematical Journal
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    • v.54 no.3
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    • pp.501-509
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    • 2014
  • In 2000, K. Denecke and K. Mahdavi showed that there are many idempotent elements in $Hyp_{N_{\varphi}}(V)$ the set of normal form hypersubstitutions of type ${\tau}=(2)$ which are not idempotent elements in Hyp(2) the set of all hypersubstitutions of type ${\tau}=(2)$. They considered in which varieties, idempotent elements of Hyp(2) are idempotent elements of $Hyp_{N_{\varphi}}(V)$. In this paper, we study the similar problems on the set of all generalized hypersubstitutions of type ${\tau}=(2)$ and the set of all normal form generalize hypersubstitutions of type ${\tau}=(2)$ and determine the order of normal form generalize hypersubstitutions of type ${\tau}=(2)$.

Skin elasticity improvement effect of Young persimmon and Heated young persimmon by decreased Advanced glycation end products(AGEs) (떫은감과 포제 떫은감의 최종당화산물 생성 억제를 통한 피부 탄력 개선 효과)

  • Kim, Soo Hyun;Lee, AhReum;Kim, SuJi;Kim, Kyeong Jo;Kwon, Ojun;Choi, Joon Young;Koo, Jin Suk;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.32 no.4
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    • pp.17-24
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    • 2017
  • Objectives : Advanced glycation end products (AGEs) is bind formation of glucose and protein. Acceleration of AGE formation during hyperglycemia is associated with the pathogenesis of diabetic complications and causes kidney and skin damage. The aim of this study was investigated the AGEs inhibitory activity and antioxidant activity of water extracts from young persimmon (YP) and heated young persimmon (HYP). Methods : Paeoniae Radix Alba (YP) is prepared by heating with 30% ethanol. AGEs formation inhibitory activities of YP and HYP measured using bovine serum albumin. To evaluate the protective effects of YP and HYP in diabetic rats induced with streptozotocin (STZ) and methyl glyoxal (MGO), SD rats were distributed into four groups; normal mice (Nor), AGEs-induced rats (Con), AGEs-induced rats treated with 100 mg/kg YP (YP), AGEs-induced rats treated with 100 mg/kg heated YP (HYP) for 3weeks. Heated young persimmon respectively decrease AGEs construction. Results : YP and HYP administration inhibited the biomarkers of AGEs in serum, kidney and skin tissues. AGE-induced rats revealed that the significant decreased collagen however, heat processing methods of young persimmon up regulated inhibits AGEs-induced collagen decrease. The expressions of AGEs were decreased in YP and HYP treated group compared with the control group in tissues. It specifies that HYP has potential to serve as a positive regulator of via AGEs path way. Conclusion : It has proposed that may have an improvement effect on diabetic complications, heated young persimmon has AGEs inhibitory excellent activities and antioxidant effect.

Hypercube Diagnosis Algorithm using Syndrome Analysis of RGC-Ring (RGC-링의 신드롬 분석을 이용한 하이퍼큐브 진단 알고리즘)

  • Kim Dong-Kun;Cho Yoon-Ki;Lee Kyung-Hee;Rhee Chung-Sei
    • Journal of KIISE:Computer Systems and Theory
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    • v.33 no.1_2
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    • pp.105-109
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    • 2006
  • Hypercube has a regular and hierarchical structure, therefore it can be applied to the development of efficient diagnosis algorithm. Kranakis and Pelc [7] have proposed HYP-DIAG algorithm to implement different method of HADA/IHADA and adaptive cube partition method after embedding the small size of ring that includes all the faulty nodes. In this paper, we propose new method to reduce testing rounds by analyzing the syndrome of RGC-rings gained in the first step of HYP-DIAG and analyze the proposed algorithm.

Relationship Between Depression Anxiety Stress Scale (DASS) and Urinary Hydroxyproline and Proline Concentrations in Hospital Workers

  • Lee, Keou-Won;Kim, Soo-Jeong;Park, Jae-Beom;Lee, Kyung-Jong
    • Journal of Preventive Medicine and Public Health
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    • v.44 no.1
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    • pp.9-13
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    • 2011
  • Objectives: Although increased reactive oxygen species (ROS) is caused by stress accelerates collagen degradation, there was no data on the relationship between stress and urinary hydroxyproline (Hyp) and proline (Pro), a good marker of collagen degradation. The purpose of this study was to evaluate the relationship between depression, anxiety, and stress (DAS) and concentrations of urinary Hyp and Pro. Methods: 97 hospital employees aged 20 to 58 were asked to fill out comprehensive self-administrated questionnaires containing information about their medical history, lifestyle, length of the work year, shit-work and DAS. Depression Anxiety Stress Scale (DASS) was applied to evaluate chronic mental disorders. Urine samples were analyzed using High Performance Liquid Chromatography (HPLC) with double derivatization for the assay of hydroxyproline and proline. Results: The mean value of Hyp and Pro concenturation in all subjects was $194.1{\pm}113.4\;{\mu}mol/g$ and $568.2{\pm}310.7\;{\mu}mol/g$. DASS values and urinary Pro concentrations were differentiated by sex (female > male, p < 0.05) and type of job (nurse > others, p < 0.05). In the stepwise multiple linear regressions, urinary Hyp and Pro concentrations were influenced by stress (Adjusted $r^2$ = 0.051) and anxiety and job (Adjusted $r^2$ = 0.199), respectively. Conclusions: We found that stress and anxiety were correlated with urinary Hyp and Pro concentrations. To identifying a definite correlation, further study in large populations will be needed.

Detect of Hypericin (HyH) gene in Hypericum erectum in Korea and Comparison of H. perforatum in Europe (한국내 고추나물의 하이퍼리신 유전자(HyH)의 탐색과 유럽의 서양고추나물과 비교)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1034-1038
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    • 2007
  • Hypericin (HyH) is a substance which is isolated a medicinal herb, Hypericum perforatum L., commonly known as St. John's Wort. Hypericum erectum is a long-lived herb that is distributed in Korea. Cloned HyH genes H. erectum of were conformed by sequencing. The cDNA Hyp-1 sequence has 732 bp with an open reading frame of 567. Thus coding for a protein of 152 amino acid residues. A BLAST re-search using the deduced nucleotide sequences in HyH gene produced significant alignments with the H. perforatum. Sequences in HyH gene showed significant homology with Rubus idaeus putative allergen Rub-i-1 mRNA, Protein sequence comparisons revealed significant homology between Hyp-1 and the phenolic oxidative coupling protein hyp-1 of H. perforatum (98%). Additionally, Hyp-1 showed sig-nificant homology with various other classes of allergens, including Pru-av-1 (62%) from Prunus avium and allergen Bet-vl-Sc3 from Betula pendula (60%). Thus, the result of this study may offer an important information to establish an assay system for chemicals of the herbal medicines for H. erectum as well as H. perforatum.

Retinoic acid loaded with chitosan nanoparticles improves spermatogenesis in scrotal hyperthermia in mice

  • Fatemeh Mazini;Mohammad-Amin Abdollahifar;Hassan Niknejad;Asma Manzari-Tavakoli;Mohsen Zhaleh;Reza Asadi-Golshan;Ali Ghanbari
    • Clinical and Experimental Reproductive Medicine
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    • v.50 no.4
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    • pp.230-243
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    • 2023
  • Objective: High temperatures can trigger cellular oxidative stress and disrupt spermatogenesis, potentially leading to male infertility. We investigated the effects of retinoic acid (RA), chitosan nanoparticles (CHNPs), and retinoic acid loaded with chitosan nanoparticles (RACHNPs) on spermatogenesis in mice induced by scrotal hyperthermia (Hyp). Methods: Thirty mice (weighing 25 to 30 g) were divided into five experimental groups of six mice each. The groups were as follows: control, Hyp induced by a water bath (43 ℃C for 30 minutes/day for 5 weeks), Hyp+RA (2 mg/kg/day), Hyp+CHNPs (2 mg/kg/72 hours), and Hyp+RACHNPs (4 mg/kg/72 hours). The mice were treated for 35 days. After the experimental treatments, the animals were euthanized. Sperm samples were collected for analysis of sperm parameters, and blood serum was isolated for testosterone measurement. Testis samples were also collected for histopathology assessment, reactive oxygen species (ROS) evaluation, and RNA extraction, which was done to compare the expression levels of the bax, bcl2, p53, Fas, and FasL genes among groups. Additionally, immunohistochemical staining was performed. Results: Treatment with RACHNPs significantly increased stereological parameters such as testicular volume, seminiferous tubule length, and testicular cell count. Additionally, it increased testosterone concentration and improved sperm parameters. We observed significant decreases in ROS production and caspase-3 immunostaining in the RACHNP group. Moreover, the expression levels of bax, p53, Fas, and FasL significantly decreased in the groups treated with RACHNPs and RA. Conclusion: RACHNPs can be considered a potent antioxidative and antiapoptotic agent for therapeutic strategies in reproductive and regenerative medicine.