• Title/Summary/Keyword: OPA1

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Stability Test and Quantitative and Qualitative Analyses of the Amino Acids in Pharmacopuncture Extracted from Scolopendra subspinipes mutilans

  • Cho, GyeYoon;Han, KyuChul;Yoon, JinYoung
    • Journal of Pharmacopuncture
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    • v.18 no.1
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    • pp.44-55
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    • 2015
  • Objectives: Scolopendra subspinipes mutilans (S. subspinipes mutilans) is known as a traditional medicine and includes various amino acids, peptides and proteins. The amino acids in the pharmacopuncture extracted from S. subspinipes mutilans by using derivatization methods were analyzed quantitatively and qualitatively by using high performance liquid chromatography (HPLC) over a 12 month period to confirm its stability. Methods: Amino acids of pharmacopuncture extracted from S. subspinipes mutilans were derived by using O-phthaldialdehyde (OPA) & 9-fluorenyl methoxy carbonyl chloride (FMOC) reagent and were analyzed using HPLC. The amino acids were detected by using a diode array detector (DAD) and a fluorescence detector (FLD) to compare a mixed amino acid standard (STD) to the pharmacopuncture from centipedes. The stability tests on the pharmacopuncture from centipedes were done using HPLC for three conditions: a room temperature test chamber, an acceleration test chamber, and a cold test chamber. Results: The pharmacopuncture from centipedes was prepared by using the method of the Korean Pharmacopuncture Institute (KPI) and through quantitative analyses was shown to contain 9 amino acids of the 16 amino acids in the mixed amino acid STD. The amounts of the amino acids in the pharmacopuncture from centipedes were 34.37 ppm of aspartate, 123.72 ppm of arginine, 170.63 ppm of alanine, 59.55 ppm of leucine and 57 ppm of lysine. The relative standard deviation (RSD %) results for the pharmacopuncture from centipedes had a maximum value of 14.95% and minimum value of 1.795% on the room temperature test chamber, the acceleration test chamber and the cold test chamber stability tests. Conclusion: Stability tests on and quantitative and qualitative analyses of the amino acids in the pharmacopuncture extracted from centipedes by using derivatization methods were performed by using HPLC. Through research, we hope to determine the relationship between time and the concentrations of the amino acids in the pharmacopuncture extracted from centipedes.

Strength Evaluation of Pinus rigida Miller Wooden Retaining Wall Using Steel Bar (Steel Bar를 이용한 리기다소나무 목재옹벽의 내력 평가)

  • Song, Yo-Jin;Kim, Keon-Ho;Lee, Dong-Heub;Hwang, Won-Joung;Hong, Soon-Il
    • Journal of the Korean Wood Science and Technology
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    • v.39 no.4
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    • pp.318-325
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    • 2011
  • Pitch pine (Pinus rigida Miller) retaining walls using Steel bar, of which the constructability and strength performance are good at the construction site, were manufactured and their strength properties were evaluated. The wooden retaining wall using Steel bar was piled into four stories stretcher and three stories header, which is 770 mm high, 2,890 mm length and 782 mm width. Retaining wall was made by inserting stretchers into Steel bar after making 18 mm diameter of holes at top and bottom stretcher, and then stacking other stretchers and headers which have a slit of 66 mm depth and 18 mm width. The strength properties of retaining walls were investigated by horizontal loading test, and the deformation of structure by image processing (AlCON 3D OPA-PRO system). Joint (Type-A) made with a single long stretcher and two headers, and joint (Type-B) made with two short stretchers connected with half lap joint and two headers were in the retaining wall using Steel bar. The compressive shear strength of joint was tested. Three replicates were used in each test. In horizontal loading test the strength was 1.6 times stronger in wooden retaining wall using Steel bar than in wooden retaining wall using square timber. The timber and joints were not fractured in the test. When testing compressive shear strength, the maximum load of type-A and Type-B was 130.13 kN and 130.6 kN, respectively. Constructability and strength were better in the wooden retaining wall using Steel bar than in wooden retaining wall using square timber.

Assessment of Genetic Variability in Two North Indian Buffalo Breeds Using Random Amplified Polymorphic DNA (RAPD) Markers

  • Sodhi, M.;Mukesh, M.;Anand, A.;Bhatia, S.;Mishra, B.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1234-1239
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    • 2006
  • Murrah and NiliRavi are the important North Indian buffalo breeds occupying the prominent position of being the highest milk producers. These breeds are more or less similar at morphological as well as physiological levels. The technique of RAPD-PCR was applied in the present study to identify a battery of suitable random primers to detect genetic polymorphism, elucidation of the genetic structure and rapid assessment of the differences in the genetic composition of these two breeds. A total of 50 random primers were screened in 24 animals each of Murrah and NiliRavi buffaloes to generate RAPD patterns. Of these, 26 (52%) primers amplified the buffalo genome generating 263 reproducible bands. The number of polymorphic bands for the 26 chosen RAPD primers varied from 3 (OPG 06 and B4) to 26 (OPJ 04) with an average of 10.1 bands per primer and size range of 0.2 to 3.2 kb. DNA was also pooled and analyzed to search for population specific markers. Two breed specific RAPD alleles were observed in each of Murrah (OPA02 and OPG16) and NiliRavi (OPG09) DNA pools. RAPD profiles revealed that 11 (4.2%) bands were common to all the 48 individuals of Murrah and NiliRavi buffaloes. Pair-wise band sharing calculated among the individual animals indicated considerable homogeneity of individuals within the breeds. Within breed, band sharing values were relatively greater than those of interbreed values. The low genetic distance (Nei's) value (0.109) estimated in this study is in accordance with the origin and geographical distribution of these breeds. The RAPD analysis indicated high level of genetic similarity between these two important North Indian buffalo breeds.

OIP5 is a highly expressed potential therapeutic target for colorectal and gastric cancers

  • Chun, Ho-Kyung;Chung, Kyung-Sook;Kim, Hee-Cheol;Kang, Jung-Eun;Kang, Min-Ah;Kim, Jong-Tae;Choi, Eun-Hwa;Jung, Kyeong-Eun;Kim, Moon-Hee;Song, Eun-Young;Kim, Seon-Young;Won, Mi-Sun;Lee, Hee-Gu
    • BMB Reports
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    • v.43 no.5
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    • pp.349-354
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    • 2010
  • Previously, we reported that overexpression of Opa (Neisseria gonorrhoeae opacity-associated)-interacting protein 5 (OIP5) caused multi-septa formation and growth defects, both of which are considered cancer-related phenotypes. To evaluate OIP5 as a possible cancer therapeutic target, we examined its expression level in 66 colorectal cancer patients. OIP5 was upregulated about 3.7-fold in tumors and over 2-fold in 58 out of 66 colorectal cancer patients. Knockdown of OIP5 expression by small interfering RNA specific to OIP5 (siOIP5) resulted in growth inhibition of colorectal and gastric cancer cell lines. Growth inhibition of SNU638 by siOIP5 caused an increase in sub-G1 DNA content, as measured by flow cytometry, as well as an apoptotic gene expression profile. These results indicate that knockdown of OIP5 may induce apoptosis in cancer cells. Therefore, we suggest that OIP5 might be a potential cancer therapeutic target, although the mechanisms of OIP5-induced carcinogenesis should be elucidated.

Zic3z Defines the Dorsal and Vegetal Neuroectoderm in the Zebrafish Embryonic Development

  • Lee, Kyu-Sun;Huh, Tae-Lin;Lee, Chang-Joong;Rhee, Myung-Chull
    • Animal cells and systems
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    • v.12 no.1
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    • pp.23-33
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    • 2008
  • The Zic family is a group of genes encoding zinc finger proteins that are highly expressed in the mammalian cerebellum. Zic genes are the vertebrate homologue of Drosophila pair-rule gene, odd-paired(opa), which plays important roles in the parasegmental subdivision as well as in the visceral mesoderm development of Drosophila embryos. Recent studies on human, mouse, frog, fish and ascidian Zic homologues support that Zic genes are involved in a variety of developmental processes, including neurogenesis, myogenesis, skeletal patterning, and left-right axis establishment. In an effort to explore possible functions of Zic proteins during vertebrate embryogenesis, we initially examined more detailed expression pattern of zebrafish homologue of zic3(zic3z). zic3z transcripts are detected in the neuroectoderm, neural plate, dorsal neural tube, and brain regions including eye field during early embryonic development. Marker DNA studies found that zic3z transcription is modulated by BMP, Wnt, and Nodal signals particularly in the dorsal and vegetal neuroectoderm at gastrula. Interfering with zic3z translation with zic3z-specific morpholino causes abnormal brain formation and expansion of the optic stalk cells. Retinal ganglion cells(RGCs) undergo abnormal neuronal differentiation. These findings suggest that zic3z defines the dorsal and vegetal neuroectoderm to specify brain formation and retinal neurogenesis during early embryonic development.

Isolation of an Acid-Labile Gene from the Seaweed Porphyra yezoensis Tissue (해조류 김 Porphyra yezoensis 엽체로부터 산에 민감한 유전자의 분리)

  • Jin, Hyung-Joo;Park, Sun-Mee;Kim, Long-Guo;Jin, Deuk-Hee;Kong, In-Soo;Hong, Yong-Ki
    • KSBB Journal
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    • v.14 no.6
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    • pp.702-706
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    • 1999
  • The genetic responses of aquaculturable seaweed Prophyra yezoensis tissue by acid shock have been compared using differential display technique. The tissue has been challenged in seawater containing 0.05% hydrogen chloride(pH 3.0) for 5 min, then rehabilitated in normal seawater for 10 min, 30 min, 60 min and 4 hrs, respectively. Total RNA was extracted by LiCl-guanidinium method. The cDNA was synthesized by reverse transcription with random hexamers and amplified by PCR with arbitrary primers. The genetic fragment disappeared by acid shock was selectively isolated from agarose gel and sequenced with a DNA auto sequencer. One of the acid-labile gene(605 bp) was identified as a dethiobiotin synthetase gene according to sequence alignment analysis by the NCBI BLAST search program.

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Micropropagation and RAPD Analysis of Somaclonal Variants in Lavandula spica cv. Marino (라벤다의 기내증식과 RAPD에 의한 체세포 변이체 분석)

  • Li, Xian Ri;Seong, Eun-Soo;Kim, Il-Seop;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.2
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    • pp.94-100
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    • 1999
  • To establish the mass propagation system of Lavandula spica cv. Marino, shoot tip, node, internode and leaf segment cultures were carried out. RAPD was applied to detect the somaclonal variation. Callus induction was very high in the medium supplemented with 1 mg/l 2.4-D, 2 mg/l NAA. especially and combined with 0.05 mg/l BAP from leaves. Shoot formation was high with $2{\sim}4\;mg/l$ BAP or 4 mg/l BAP + 0.2 mg/l NAA from shoot tip. Shoot proliferation was 9.1 times in the $B_{5}$ medium with 0.5 mg/l BAP and 0.01 mg/l NAA. Root formation was improved in NAA, which was the concentration of 0.1 to 1 mg/l and 1 mg/l IAA. Nursery survival rate was enhanced over 90% and growth was looked good in the acclimation soil consisting of peatmoss : vermiculite : perlite (1:1:1, v:v:v). Randomly amplified polymorphic DNA banding patterns based on polymerase chain reaction (PCR) were used to assess the genetic variation in plants regenerated from in vitro culture.

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Augmenter of Liver Regeneration Alleviates Renal Hypoxia-Reoxygenation Injury by Regulating Mitochondrial Dynamics in Renal Tubular Epithelial Cells

  • Long, Rui-ting;Peng, Jun-bo;Huang, Li-li;Jiang, Gui-ping;Liao, Yue-juan;Sun, Hang;Hu, Yu-dong;Liao, Xiao-hui
    • Molecules and Cells
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    • v.42 no.12
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    • pp.893-905
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    • 2019
  • Mitochondria are highly dynamic organelles that constantly undergo fission and fusion processes that closely related to their function. Disruption of mitochondrial dynamics has been demonstrated in acute kidney injury (AKI), which could eventually result in cell injury and death. Previously, we reported that augmenter of liver regeneration (ALR) alleviates renal tubular epithelial cell injury. Here, we gained further insights into whether the renoprotective roles of ALR are associated with mitochondrial dynamics. Changes in mitochondrial dynamics were examined in experimental models of renal ischemia-reperfusion (IR). In a model of hypoxia-reoxygenation (HR) injury in vitro, dynamin-related protein 1 (Drp1) and mitochondrial fission process protein 1 (MTFP1), two key proteins of mitochondrial fission, were downregulated in the Lv-ALR + HR group. ALR overexpression additionally had an impact on phosphorylation of Drp1 Ser637 during AKI. The inner membrane fusion protein, Optic Atrophy 1 (OPA1), was significantly increased whereas levels of outer membrane fusion proteins Mitofusin-1 and -2 (Mfn1, Mfn2) were not affected in the Lv-ALR + HR group, compared with the control group. Furthermore, the mTOR/4E-BP1 signaling pathway was highly activated in the Lv-ALR + HR group. ALR overexpression led to suppression of HR-induced apoptosis. Our collective findings indicate that ALR gene transfection alleviates mitochondrial injury, possibly through inhibiting fission and promoting fusion of the mitochondrial inner membrane, both of which contribute to reduction of HK-2 cell apoptosis. Additionally, fission processes are potentially mediated by promoting tubular cell survival through activating the mTOR/4E-BP1 signaling pathway.

Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains with higher β-glucan (베타글루칸 함량이 높은 큰느타리버섯 선발을 위한 SCAR marker 개발)

  • Kim, Su Cheol;Kim, Hye Soo;Cho, Yong Un;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.79-83
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    • 2015
  • In this study, SCAR marker that differentiates Pleurotus eryngii strains with higher ${\beta}$-glucan from control strain was developed. Genomic DNAs of 9 control strains of Pleurotus eryngii and 9 Pleurotus eryngii strains with higher ${\beta}$-glucan were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). One-hundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 91 bp was yielded by OP-R03 primer from the Pleurotus eryngii strains with higher ${\beta}$-glucan. A sequence characterized amplified region (SCAR) marker, designated as OP-R03-1-F and OP-R03-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-R03-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains with higher ${\beta}$-glucan from the control strains.

Comparison of Dissolved Ammonium Analytical Method in Seawater: Spetrophotometry and Fluorometry (해수 중 용존 암모늄 분석방법 비교: 분광광도법과 형광법)

  • SON, PURENA;PARK, JOONSEONG;RHO, TAEKEUN
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.25 no.4
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    • pp.81-96
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    • 2020
  • Berthlot's reaction spectrophotometric method is generally used for the analysis of dissolved ammonium in seawater, but in recent years, a fluorescence method using an orthophthaldialdehyde-sulfite (OPA) fluorescent reagent is actively used internationally. In this study, we investigated the effects of the detection limit between the analysis methods, the reagent refractive index inherent in the spectrophotometric method, and the use of different calibration curves to understand the cause of the difference in dissolved ammonium concentration (about 0.31 𝜇M) observed in the seawater samples and a nutrient reference material between two institutions (KIOST (spectrophotometric method, one-order linear regression gradient only), Australia CSIRO (fluorescence method, quadratic formula)) conducted onboard the Australian R/V Investigator in 2017. The method detection limit (0.063 𝜇M) and the reagent refractive index background value (0.054 𝜇M) of the spectrophotometric method measured in this study could explain the difference in dissolved ammonium concentration values of the two institutes about 20% and 17%, respectively. However, when the concentration of the calibration curve of the spectrophotometric method was calculated using the same quadratic as the fluorescence method or the slope and intercept of linear regression, the difference in the dissolved ammonium concentration between the two institutions was reduced to less than the detection limit of the spectrophotometric method. Therefore, the difference in the concentration of dissolved ammonium between the two institutions, found in the nutrient reference materials and the seawater field sample during the international onboard nutrient inter-comparison experiment, may be attributed to be the effect of the different calibration curves used in the two methods rather than the effect of the difference in two analytical methods. When comparing the dissolved ammonium data from seawater samples in the future, it is recommended to pay attention to the information on the baseline, number of standard solutions, and calibration curve used in the analysis.