• 제목/요약/키워드: 25-Hydroxylase

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Cyclosporin A-induced Gingival Overgrowth is Closely Associated with Regulation Collagen Synthesis by the Beta Subunit of Prolyl 4-hydroxylase and Collagen Degradation by Testican 1-mediated Matrix Metalloproteinase-2 Expression

  • Park, Seong-Hee;Kim, Jae-Yoen;Kim, Hyun-Jeong;Park, Kwang-Kyun;Cho, Kyoo-Sung;Choi, Seong-Ho;Chung, Won-Yoon
    • International Journal of Oral Biology
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    • v.33 no.4
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    • pp.205-211
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    • 2008
  • Gingival overgrowth can cause dental occlusion and seriously interfere with mastication, speech, and dental hygiene. It is observed in 25 to 81% of renal transplant patients treated with cyclosporine A (CsA). CsA-induced gingival overgrowth (CIGO) is caused by quantitative alteration of the extracellular matrix components, particularly collagen. However, the molecular mechanisms involved in the pathogenesis of CIGO remain poorly understood, despite intense clinical and laboratory investigations. The aim of the present work is to identify differentially expressed genes closely associated with CIGO. Human gingival fibroblasts were isolated by primary explant culture of gingival tissues from five healthy subjects (HGFs) and two patients with the CIGO (CIGO-HGFs). The proliferative activity of CsA-treated HGFs and CIGO-HGFs was examined using the MTT assay. The identification of differentially expressed genes in CsA-treated CIGO-HGF was performed by differential display reverse transcriptase-polymerase chain reaction (RT-PCR) followed by DNA sequencing. CsA significantly increased the proliferation of two HGFs and two CIGO-HGFs, whereas three HGFs were not affected. Seven genes, including the beta subunit of prolyl 4-hydroxylase (P4HB) and testican 1, were upregulated by CsA in a highly proliferative CIGO-HGF. The increased P4HB and testican-1 mRNA levels were confirmed in CsA-treated CIGO-HGFs by semiquantitative RT-PCR. Furthermore, CsA increased type I collagen mRNA levels and suppressed MMP-2 mRNA levels, which are regulated by P4HB and testican-1, respectively. These results suggest that CsA may induce gingival overgrowth through the upregulation of P4HB and testican-1, resulting in the accumulation of extracellular matrix components.

MALDI-MS-Based Quantitative Analysis of Bioactive Forms of Vitamin D in Biological Samples

  • Ahn, Da-Hee;Kim, Hee-jin;Kim, Seong-Min;Jo, Sung-Hyun;Jeong, Jae-Hyun;Kim, Yun-Gon
    • Korean Chemical Engineering Research
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    • v.58 no.1
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    • pp.106-112
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    • 2020
  • Analyzing vitamin D levels is important for monitoring health conditions because vitamin D deficiency is associated with various diseases such as rickets, osteomalacia, cardiovascular disorders and some cancers. However, vitamin D concentration in the blood is very low with optimal level of 75 nmol/L, making quantitative analysis difficult. The objective of this study was to develop a highly sensitive analysis method for vitamin D using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS). 25-hydroxyvitamin D (25(OH)D), which has been used as an indicator of vitamin D metabolites in human biofluids was chemically derivatized using a secosteroid signal enhancing tag (SecoSET) with powerful dienophile and permanent positive charge. The SecoSET-derivatized 25(OH)D provided good linearity (R2 > 0.99) and sensitivity (limit of quantitation: 11.3 fmol). Chemical derivatization of deuterated 25-hydroxyvitamin D3 (d6-25(OH)D3) with SecoSET enabled absolute quantitative analysis using MALDI-MS. The highly sensitive method could be successfully applied into monitoring of quantitative changes of bioactive vitamin D metabolites after treatment with ketoconazole to inhibit 1α-hydroxylase reaction related to vitamin D metabolism in human breast cancer cells. Taken together, we developed a MALDI-MS-based platform that could quantitatively analyze vitamin D metabolites from cell products, blood and other biofluids. This platform may be applied to monitor various diseases associated with vitamin D deficiency such as rickets, osteomalacia and breast cancer.

Korean Red Ginseng protects dopaminergic neurons by suppressing the cleavage of p35 to p25 in a Parkinson's disease mouse model

  • Jun, Ye Lee;Bae, Chang-Hwan;Kim, Dongsoo;Koo, Sungtae;Kim, Seungtae
    • Journal of Ginseng Research
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    • v.39 no.2
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    • pp.148-154
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    • 2015
  • Background: Ginseng is known to have antiapoptotic, anti-inflammatory, and antioxidant effects. The present study investigated a possible role of Korean Red Ginseng (KRG) in suppressing dopaminergic neuronal cell death and the cleavage of p35 to p25 in the substantia nigra (SN) and striatum (ST) using a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson's disease mouse model. Methods: Ten-week-old male C57BL/6 mice were injected intraperitoneally with 30 mg/kg of MPTP at 24-h intervals for 5 d, and then administered KRG (1 mg/kg, 10 mg/kg, or 100 mg/kg) once a day for 12 consecutive days from the first injection. Pole tests were performed to assess the motor function of the mice, dopaminergic neuronal survival in the SN and ST was evaluated using tyrosine hydroxylase-immunohistochemistry, and the expressions of cyclin-dependent kinase 5 (Cdk5), p35, and p25 in the SN and ST were measured using Western blotting. Results: MPTP administration caused behavioral impairment, dopaminergic neuronal death, increased Cdk5 and p25 expression, and decreased p35 expression in the nigrostriatal system of mice, whereas KRG dose-dependently alleviated these MPTP-induced changes. Conclusion: These results indicate that KRG can inhibit MPTP-induced dopaminergic neuronal death and suppress the cleavage of p35 to p25 in the SN and the ST, suggesting a possible role for KRG in the treatment of Parkinson's disease.

Estimation of Death Time by Changes of Postmortem Xanthine Oxidase Activity in Rats

  • Yoon, Hyung-Won;Yoon, Chong-Guk;Cho, Hyun-Gug
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.439-442
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    • 2006
  • To evaluate the postmortem changes in activities of oxygen free radical metabolizing enzymes, the rats were sacrificed with cervical dislocation and were kept in an incubator at $25^{\circ}C$, 70% of humidity for 12 hours. The activities of aniline hydroxylase, catalase, glutathione-S-transferase and superoxlde dismutase were decreased with the time. On the other hand, the activity and type conversion ratio (type D ${\to}$type O) of hepatic xanthine oxidase (XO) were gradually increased. From these changes of XO, the estimation of death time (mathematical equation) could be determined with the least square method. To clarify the cause of increasing XO activity, enzyme kinetics were examined. The Km values of XO were decreased with the time. In conclusion, the determination of liver XO activity might be used for the estimation of death time in the early postmortem period.

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Effect of Cyclohexane Application to Rat Skin on the Skin Toxicity (흰쥐의 피부조직에 있어서 Cyclohexane의 독성)

  • 전태원;조현국;윤종국
    • Journal of Environmental Health Sciences
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    • v.28 no.2
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    • pp.71-80
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    • 2002
  • To evaluate the skin toxicity of topical cyclohexane application (25mg/$\textrm{cm}^2$) was sequentially applied to the rat skin for four days. On the histopathological findings in the light micrographs, neutrophils and engulfed neutrophils are seen, and many cytoplasmic processes were appeared in proliferated layer whereas in the dermis area, increased numbers of fibroblast, accumulation of neutrophil and lipid droplets are demonstrated. On the other hand, applying the cyclohexane to the rat skin led to the remarkable rise of cutaneous xanthine oxidase activity and similar activities of superoxide dismutase and glutathione peroxidase and glutathione content and declined activity of glutathione S-transferase compared with control group. Especially the remarkably decreased activity of aniline hydroxylase (AH) was appeared in skin as little as scarcely determined. Furthermore, the applying the cyclohexane to skin led to the significantly increased activity of hepatic AH and alcohol dehydrogenase. These results indicate that oxygen free radical and intermediate metabolite of cyclohexane may be responsible for structural changes in skin by cyclohexane application to rat skin.

Bioconversion of progesterone by immobilized aspergillus phoenicis (고정화된 aspergillus phoenicis를 이용한 progesterone 전환)

  • 박희은;김말남
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.70-76
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    • 1989
  • Progestrone bioconversion by immobilized Aspergillus phoenicis was studied. Progesterone was converted into 11$\alpha$-hydroxyprogesterone and 3-minor byproducts. Whole cells of A. phoenicis were immobillized by enreappment with calcium-alginate, K-carrageenan, or polyacrylamide. Of these materials tested, cell immobilized in $Ca^{2+}$ -alginate gels showed the highest activity for 11$\alpha$-hydroxylation of progesterone. In the case of mycelia immobilized in $Ca^{2+}$-alginate, futher progressing hydroxylation of 11$\alpha$-hydroxyprogesterone was greatly reduced. Spores of A. phoenicis which were immobillized with $Ca^{2+}$-alginate and germinatedin situ for 25 hours showed higher 11$\alpha$-hydroxylase activity than those of entrapped whole mycelia and maintained initial enzyme activity for all 8 times of repeated use. After 16 times of reuse, the activity was declined 30% or more. When culture media and $Zn^{2+}$ were introduced into the reaction media, the activity of the immobilized mycelia which had been lowered due to many times of reuse was effectively reactivated.

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Effect of the Repeated Treatment of Xylene to the Rats on the Xylene Metabolism (흰쥐에 Xylene반복 투여가 Xylene의 대사에 미치는 영향)

  • 이혜자;조현국;이상일;전태원;윤종국
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.59-66
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    • 1999
  • To evaluate the effect of repeated treatment of xylene on its metabolism, m-xylene (0.25 ml of 50% in olive oi1/100 g body weight) has been intraperitoneally given to the rats 1, 4, 8, 12 and 16 times every other day. m-Xylene was once more administered to the animals after 24 hrs since last injection of it. And then the animals were sacrificed after 24 hrs. Four times xylene treated rats showed the significantly elevated urinary m-methylhippuric acid, compared to those treated with the single dose of m-xylene with the continued similiar high levels of urinary m-methylhippuric acid up to the animals pretreated 12 times and then those treated 16 times defined the significantly decreased urinary m-methylhippuric acid compared to those treated 12 times. On the other hand, hepatic aniline hydroxylase and alcohol dehydrogenase activities demonstrated a gradual increase from the first group to the 12 times xylene-treated animals, but those treated 16 times showed the significantly decreased value compared with the 12 times treated-group. And aldehyde dehydrogenase activities in rats treated with m-xylene 8, 12 or 16 times were significantly decreased compared to those pretreated one or four times. In the early stage of xylene administration, proliferation of SERs were seen whereas SERs were decreased and RERs were clearly increased in xylene-treated rats 16 times. These results indicate that the frequency of xylene injection may influence upon the changes in xylene metabolite, m-methylhippuric acid and it may be due to induction of xylene metabolizing enzymes.

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Lack of Associations between Vitamin D Metabolism-Related Gene Variants and Risk of Colorectal Cancer

  • Mahmoudi, Touraj;Karimi, Khatoon;Arkani, Maral;Farahani, Hamid;Nobakht, Hossein;Dabiri, Reza;Asadi, Asadollah;Vahedi, Mohsen;Zali, Mohammad Reza
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.957-961
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    • 2014
  • Purpose: With regard to the protective effect of vitamin D against colorectal cancer (CRC), we evaluated genetic variants that might influence vitamin D metabolism: vitamin D receptor (VDR), vitamin D binding protein (GC), vitamin D 25-hydroxylase (CYP2R1), and vitamin D 25-hydroxy 1-alpha hydroxylase (CYP27B1). Materials and Methods: A total of 657 subjects, including 303 cases with CRC and 354 controls were enrolled in this case-control study. All 657 were genotyped for the four gene variants using PCR-RFLP methods. Results: In this study, no significant difference was observed for VDR (rs2238136), GC (rs4588), CYP2R1 (rs12794714), and CYP27B1 (rs3782130) gene variants in either genotype or allele frequencies between the cases with CRC and the controls and this lack of difference remained even after adjustment for age, BMI, sex, smoking status, NSAID use, and family history of CRC. Furthermore, no evidence for effect modification of the variants and CRC by BMI, sex, or tumor site was observed. Conclusions: Our findings do not support a role for VDR, GC, and CYP27B1 genes in CRC risk in our Iranian population. Another interesting finding, which to our knowledge has not been reported previously, was the lack of association with the CYP2R1 gene polymorphism. Nonetheless, our findings require confirmation and possible roles of vitamin D metabolism-related genes in carcinogenesis need to be further investigated.

A Case of Type I Vitamin D-dependent Rickets with Unilateral Aplasia of Kidney (일측성 신장 무형성을 동반한 제 1형 비타민 D 의존성 구룻병 1례)

  • Lim, Dong-Hee;Jung, Ji-In;Yim, Hyung-Eun;Eun, Baik-Lin;Yoo, Kee-Hwan;Hong, Young-Sook;Lee, Joo-Won
    • Childhood Kidney Diseases
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    • v.12 no.1
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    • pp.111-115
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    • 2008
  • Vitamin D-dependent rickets(VDDR) is a rare autosomal disorder, characterized by hypocalcemia, hypophosphatemia, increased alkaline phosphatase, secondary hyperparathyroidism and many other clinical features. Type I VDDR is due to congenital defects of renal 1${\alpha}$-hydroxylase, the enzyme responsible for the conversion of 25-(OH)D3 to 1,25-$(OH)_2D3$. Type II VDDR arise from target organ resistance to 1,25-$(OH)_2D3$. Unilateral renal aplasia is generally thought to result from a lack of induction of the metanephric blastema from the ureteral bud, which may be secondary to ureteral bud maldevelopment and/or to a problem with the formation of the mesonephric duct. The incidence of unilateral renal aplasia is approximately 1/500-3,200. Type 1 VDDR associated with unilateral renal aplasia has not been reported yet. Thus we report a case of a 3 month old female infant diagnosed as type 1 VDDR with unilateral aplasia of kidney.

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Gene Expression as Related to Ripening in High Temperature during Different Coloration Stages of 'Haryejosaeng' and 'Shiranuhi' Mandarin Fruits (온주밀감 '하례조생'과 '부지화' 과실의 착색 단계별 고온에 의한 성숙 관련 유전자의 발현 변화)

  • Ahn, Soon Young;Kim, Seon Ae;Moon, Young-Eel;Yun, Hae Keun
    • Horticultural Science & Technology
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    • v.34 no.5
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    • pp.665-676
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    • 2016
  • As high temperature during citrus growing season has caused a serious problems including inferior coloration in production of mandarins in Korea, we were to investigate the expression pattern of several genes related with coloration during the ripening in high temperature condition of citrus fruits. The expression of genes related with sugar metabolism, cell wall degradation, and flavonoid synthesis in high temperature conditions was investigated in fruits of 'Haryejosaeng' (Citrus unshiu) and 'Shiranuhi' mandarin (C. reticulata). While the expression of beta-amylase (BMY), phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), and flavanone 3-hydroxylase (F3H) was differently induced, expression of polygalacturonase (PG) decreased dependently on temperature conditions. In 'Haryejosaeng' mandarin, while the expression of genes related to the skin coloration, such as CHS and F3H genes increased at $25^{\circ}C$, the expression of PAL and stilbene synthase (STS) genes were induced at $30-35^{\circ}C$ in all ripening stages. In 'Shiranuhi' mandarin, the expression of the BMY gene decreased at early time point in all temperature condition and then increased at $30-35^{\circ}C$ than at $25^{\circ}C$ in the ripening stage 2 to 3 of fruits. F3H and STS genes also showed the tendency to decrease at $30-35^{\circ}C$. Although the expression levels of genes in ripening stage 1 and stage 2 of fruits showed similar patterns in both 'Haryejosaeng' and 'Shiranuhi', the expression levels of genes were down-regulated in late ripening stage of 'Shiranuhi' fruits compared to 'Haryejosaeng'. In general, the mRNA levels of seven tested genes were higher in 'Haryejosaeng' than in 'Shiranuhi' mandarin, and expression of genes by high temperature was regulated sensitively in 'Haryejosaeng' compared to 'Shiranuhi' mandarin. Further investigations of expression of various genes based on transcriptome analysis in early ripening stage can provide valuable information about the responses to climatic changes in ripening citrus fruits.