• Title/Summary/Keyword: Stress Function

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Seminal Plasma Heparin Binding Proteins Improve Semen Quality by Reducing Oxidative Stress during Cryopreservation of Cattle Bull Semen

  • Patel, Maulikkumar;Gandotra, Vinod K.;Cheema, Ranjna S.;Bansal, Amrit K.;Kumar, Ajeet
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권9호
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    • pp.1247-1255
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    • 2016
  • Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, $40{\mu}g/mLs$ semen were standardized to find out the optimum dose and $20{\mu}g/mLs$ was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with $20{\mu}g/mL$ SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/$32.7{\mu}M$/$10^9$ spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in motility, HOST and in vitro acrosome reaction. It can be concluded that treatment of neat semen with SP-HBPs before cryopreservation minimized the cryoinjury by decreasing the generation of reactive oxygen species.

금주환자(禁酒患者) 102명(名)에 대(對)한 임상적(臨床的) 고찰(考察) (강원도지역(江原道地域)을 중심으로) (Clinical Study of the effect of Ear Acupuncture on 102 Alcoholics)

  • 강재춘
    • 대한한의학회지
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    • 제17권2호
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    • pp.318-330
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    • 1996
  • We had been investigated the effect of ear acupuncture in alcoholics, but in Kang won do province never we have been investigated. so then in Sangji university oriental hospital from Mar. 1. 1995 to Feb. 28. 1996. we clinically analized 102 alcoholics treated The results were summarized as follow: 1. In Age distribution, 40s were the top as 28.4% and next 30s, 50s, 60s 70s, below 3Os. In Sex distribution, these were 92 person in male, 10 person in female. 2. Marrige Status revealed in descending order ; married, single, divorced Sibling order distribution in descending order; eldest son, middle son, youngest son, only son 3. Distribution of education in descending order ; high school, middle school, primary school, collage and graduate school, illiterate 4. Distribution of occupation in descending order; farmer, labor, merchent, salaried and inoccupation, service 5. Distribution of religion in descending order ; none, buddism, protestantism, catholicism 6. Distribution of family history of alcoholism; yes(24.5%), no(69.6%) 7. Distribution of onset of drinking in descending order ; 20s, below 20s, 30s and 40s, 50s 8. Distribution of duration of drinking in descending order ; 20-29 years, 10-19 and below years, 40-49 years, 30-39 years 9. Distribution of the reason of drinking in descending order ; habbit, business and reduce of stress, no reason, syndrome of stop drinking 10. Distribution of frequency of drinking in descending order ; daily, four or five times a week, irregularly, once a week, two or three times a week 11. Distribution of amount of drinking in descending order ; two bottles, one bottle, three bottles, half, bottle above four bottles 12. Distribution of Chief Complaint of Alcohol in descending order ; no appetite and anorexia, diarrhrea insomnia, fatigue, vomitting, tremor, drinking water, hallucination, abdominal pain, constipation 13. Distribution of total MAST score in descending order ; 26-48 score, 13-18 score, 19-25 score, 8-12 score, 0-7 score 14. In the treatment effects according to MAST score, these were complete stop drinking 18.0%, improved 53.0%, unchanged 29.0%. 15. Distribution of liver function test and treatment effects in descending order ; ALAT, ASAT, GGTP, Trigliceride, Alk-phosphatase and Total bilirubin The treatment effects in ear acupuncture were 70.5% effective ones ; stop drinking 17.6%, improved 52.9%. 17. These were headache, nausea, vomitting sense, weakness in revealing symtoms after treatment, but no severe side effect. 18. Distribution of the times of onset in the change of taste in descending order; 3-5 times, below 2 times and above 6 times.

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Molecular analysis of alternative transcripts of equine AXL receptor tyrosine kinase gene

  • Park, Jeong-Woong;Song, Ki-Duk;Kim, Nam Young;Choi, Jae-Young;Hong, Seul A;Oh, Jin Hyeog;Kim, Si Won;Lee, Jeong Hyo;Park, Tae Sub;Kim, Jin-Kyoo;Kim, Jong Geun;Cho, Byung-Wook
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권10호
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    • pp.1471-1477
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    • 2017
  • Objective: Since athletic performance is a most importance trait in horses, most research focused on physiological and physical studies of horse athletic abilities. In contrast, the molecular analysis as well as the regulatory pathway studies remain insufficient for evaluation and prediction of horse athletic abilities. In our previous study, we identified AXL receptor tyrosine kinase (AXL) gene which was expressed as alternative spliced isoforms in skeletal muscle during exercise. In the present study, we validated two AXL alternative splicing transcripts (named as AXLa for long form and AXLb for short form) in equine skeletal muscle to gain insight(s) into the role of each alternative transcript during exercise. Methods: We validated two isoforms of AXL transcripts in horse tissues by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned the transcripts to confirm the alternative locus and its sequences. Additionally, we examined the expression patterns of AXLa and AXLb transcripts in horse tissues by quantitative RT-PCR (qRT-PCR). Results: Both of AXLa and AXLb transcripts were expressed in horse skeletal muscle and the expression levels were significantly increased after exercise. The sequencing analysis showed that there was an alternative splicing event at exon 11 between AXLa and AXLb transcripts. 3-dimentional (3D) prediction of the alternative protein structures revealed that the structural distance of the connective region between fibronectin type 3 (FN3) and immunoglobin (Ig) domain was different between two alternative isoforms. Conclusion: It is assumed that the expression patterns of AXLa and AXLb transcripts would be involved in regulation of exercise-induced stress in horse muscle possibly through an $NF-{\kappa}B$ signaling pathway. Further study is necessary to uncover biological function(s) and significance of the alternative splicing isoforms in race horse skeletal muscle.

상황버섯의 기술개발 동향과 시장분석 (Phellinus linteus; Market and Technology Trends Analysis)

  • 손은화;노현숙;박영서;손은수;강세찬;강남성;표석능
    • KSBB Journal
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    • 제23권2호
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    • pp.109-117
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    • 2008
  • 담자균류의 버섯은 대부분 영양생장단계의 균사체와 포자를 형성하는 생식생장단계인 자실체로 나누어진다. 그러나 자연산 상황버섯은 자연계에서의 분포수가 극히 적어 그 자실체를 얻기 어려우며, 이러한 희귀성으로 인해 매우 고가로 거래되고 있다. 1990년대 중반 상황버섯은 균사체 배양기술과 인공재배법이 확립된 후 농가에서 대량생산되기 시작하였으며, 이에 따라 상황버섯의 유효성분을 이용한 기능성 식품 및 의약품의 개발이 활발하였다. 그러나 인공재배법에 의한 상황버섯 추출물은 자연산 추출물의 항암 및 효능면에서 차이를 나타내고 있어, 자연산 자실체와 같은 수준에서의 약효를 나타내기 위한 인공배양기술과 고농도 유효성분을 획득하기 위한 추출법이 핵심기술로 대두되었고, 인공배양된 상황버섯과 자연산의 다각도적인 약효 비교에 관한 연구가 아직까지 진행되고 있다. 따라서 상황버섯의 유효성분이 높게 함유된 저렴한 추출물이 생산될 수 있는 기반 기술들이 기능성 음료용수, 식품 및 의약품으로 개발에 중요한 핵심기술로 떠오르고 있다. 국내 상황버섯 기능성 식품의 시장은 상황버섯의 효능에 대한 인지 증대, 웰빙열풍을 의한 건강 음료에 대한 선호도 증대 등으로 인해 향후에도 10% 이상의 높은 성장률을 보일 것으로 예상된다. 현재는 중소기업들이 주로 활동하고 있어 마케팅 및 유통의 한계로 인해 시장이 크게 확대되고 있지 않으나, 사업매력도가 긍정적으로 평가되고 있어 대기업에서도 상황버섯을 이용한 기능성 식품 시장에의 진출을 모색하고 있다.

청동풍탱이(Anomala albopilosa)추출물의 항산화성 및 생리기능 (Antioxidant Activity and Physiological Function of the Anomala albopilosa Extracts)

  • 윤원종;이정아;김지영;김상범;박수영
    • 한국식품영양과학회지
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    • 제36권6호
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    • pp.670-677
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    • 2007
  • 산화적인 스트레스(oxidative stress)는 신경염증의 발병 요인 중의 하나로 알려져 있다. 이에 본 연구는 약용곤충으로 알려진 청동풍뎅이(Anomala albopilosa) 성충과 3령 유충(궁벵이) 에탄올 추출물을 대상으로 항산화활성을 비교 측정하였으며, 또한 대식세포인 RAW 264.7 세포에서의 NO 의 생성억제 효과 및 세포독성 그리고 분자 염증관련 인자인 iNOS, COX-2와 $PGE_2$ 생성 및 활성 억제를 조사하여 항염 증효과 등의 생리활성을 측정 비교하였다. 우선, 분자염증은 활성산소 관련의 물질과 밀접한 관련이 있으므로 향산화 실험과 관련하여 청동풍탱이 성충과 3령 유충 에탄올 추출물 의 DPPH(1,1-diphenyl-2-picrylhydrazyl) radical 소거능, superoxide radical 소거능, xanthine oxidase 저해활성 그리고 nitric oxide 소거능에 대한 항산화활성 등의 assay를 실시하였다. 그 결과, 청동풍뎅이 에탄올 추출물은 항산화능을 갖고 있었으며, 3령 유충 추출물에서보다는 성충 추출물에서 다소 높은 항산화활성을 보여주었다. 또한 RAW 264.7 세포에 LPS로 자극을 주고 청동풍뎅이 성충과 3령 유충 에탄올 추출물을 처리하여 항염증관련 활성을 확인해본 결과, 고농도에서는 다소 세포독성을 나타냈지만 농도의존적으로 NO, iNOS, COX-2 그리고 $PGE_2$의 생성 억제효과가 나타났다. 이러한 결과는 유용곤충자원을 이용한 유효성분 추출을 통한 항산화 및 항염증 물질의 연구 또는 예방하거나 치료할수 있는 염증 억제 성분의 분리 및 그 작용기전 연구에 중요한 기초 자료가 될 것이라 사료된다.

횡보강근이 없는 100 MPa 이하 콘크리트의 철근 압축이음 강도와 이음길이 (Capacity and Length of Compression Lap Splice in Unconfined Concrete of 100MPa and Less Compressive Strength)

  • 천성철;이성호;오보환
    • 콘크리트학회논문집
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    • 제22권5호
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    • pp.659-666
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    • 2010
  • 현행 설계기준식에 따르면 초고강도 콘크리트에서는 철근 인장이음길이보다 압축이음길이가 더 길어지는 현상이 발생된다. 압축이음은 단부 지압이 존재하므로, 인장이음보다 길 필요는 없다. 초고강도 콘크리트의 경제적 실용화를 위해 합리적인 압축이음강도의 평가가 필요하다. 이를 위해 설계강도 80, 100 MPa 콘크리트를 이용하여 횡보강근이 없는 압축이음 실험을 수행하였다. 실험 결과 압축이음강도는 콘크리트 강도의 제곱근에 비례하는 것으로 평가되었다. 지압에 의해 발현되는 이음강도는 이음길이와 무관하며 콘크리트 강도의 제곱근에 비례하는 것으로 평가되었다. 58개 실험체의 평균은 $16.5\;\sqrt{f_{ck}}$이다. 부착에 의해 발현되는 강도를 ACI 408식으로 평가한 결과 [실험값]/[이론값]의 비는 평균 0.94로, 인장이음의 부착강도와 거의 유사하였다. 따라서 인장이음강도에 대비한 압축이음강도의 향상은 단부 지압효과로 설명될 수 있다. 실험결과에서 분석된 이음길이와 콘크리트 강도의 영향 특성을 고려하여 압축이음강도 평가식과 압축이음길이 설계식을 제안하였다. 제안된 설계식은 통계적 기법에 기반을 두어, 이음부가 철근 재료강도와 동일한 수준의 신뢰성을 확보할 수 있다.

Characterization of Mutations in AlHK1 Gene from Alternaria longipes: Implication of Limited Function of Two-Component Histidine Kinase on Conferring Dicarboximide Resistance

  • Luo, Yiyong;Yang, Jinkui;Zhu, Mingliang;Yan, Jinping;Mo, Minghe;Zhang, Keqin
    • Journal of Microbiology and Biotechnology
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    • 제18권1호
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    • pp.15-22
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    • 2008
  • Four series (S, M, R, and W) of Alternaria longipes isolates were obtained based on consecutive selection with Dimethachlon (Dim) and ultraviolet irradiation. These isolates were then characterized according to their tolerance to Dim, sensitivity to osmotic stress, and phenotypic properties. All the selected Dim-resistant isolates showed a higher osmosensitivity than the parental strains, and the last generation was more resistant than the first generation in the M, R, and W series. In addition, the changes in the Dim resistance and osmotic sensitivity were not found to be directly correlated, and no distinct morphologic characteristics were found among the resistant and sensitive isolates, with the exception of the resistant isolate K-11. Thus, to investigate the molecular basis of the fungicide resistance, a group III two-component histidine kinase (HK) gene, AlHK1, was cloned from nineteen A. longipes isolates. AlHK1p was found to be comprised of a six 92-amino-acid repeat domain (AARD), HK domain, and response regulator domain, similar to the Os-1p from Neurospora crassa. A comparison of the nucleotide sequences of the AlHK1 gene from the Dim-sensitive and -resistant isolates revealed that all the resistant isolates contained a single-point mutation in the AARD of AlHK1p, with the exception of isolate K-11, where the AlHK1p contained a deletion of 107 amino acids. Moreover, the AlHK1p mutations in the isolates of each respective series involved the same amino acid substitution at the same site, although the resistance levels differed significantly in each series. Therefore, these findings suggested that a mutation in the AARD of AlHK1p was not the sole factor responsible for A. longipes resistance to dicarboximide fungicides.

Isoproterenol 투여로 유발된 심근세포 손상에 미치는 diltiazem의 영향 (Effects of Diltiazem on Isoproterenol-induced Myocardial Cell Wounding in the Rabbit)

  • 김현;장대영;라봉진;김호덕
    • Applied Microscopy
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    • 제27권2호
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    • pp.121-130
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    • 1997
  • It has been demonstrated that majority of cells in the mammalian body such as myocytes and epithelial cells of skin and intestine respond to mechanical force or environmental factors and exhibit partial disruption of cell membrane, i. e., cell wounding, even in a physiological condition. Myocardial cells are rather apt to be wounded than other cells since they are definitely exposed to mechanical stress by contraction-relaxation and blood flow. However, the mechanism how myocardial cells protect themselves against cell wounding is not yet clarified. On this background, the present study was performed to elucidate whether albumin leakage is related to cell wounding and to assess whether diltiazem, a potent calcium channel blocker, is beneficial in isoproterenol-induced cell wounding in the heart. Hearts isolated from New Zealand White rabbits ($1.5\sim2.0kg$ body weight, n=20) were perfused with Tyrode solution by Langendorff technique. After stabilization of baseline hemodynamics, the hearts were subjected to bolus administration of isoproterenol and diltiazem as following order: $1.6{\mu}M$ isoproterenol at zero min (the beginning point): $16{\mu}M$ diltiazem at 20min; $1.6{\mu}M$ isoproterenol at 25min; $16{\mu}M$ isoproterenol at 45 min; $160{\mu}M$ diltiazem at 65 min; $16{\mu}M$ isoproterenol at 70 min. During all experiments, the left ventricular function was recorded, albumin leakage in the coronary effluents was analyzed by electrophoresis and Western blot, and myocardial cell membranes were examined by conventional transmission electron microscopy. Data were analyzed by t-test and linear regression test. Isoproterenol significantly increased the inotropic and chronotropic contractions, coronary flow, and frequency of arrhythmia, however, diltiazem did not influence on hemodynamics except decrease in the frequency of arrhythmia and a slight decrease in contractility. Isoproterenol also resulted partial disruption of myocardial cell membrane and inclose in albumin leakage, while diltiazem pretreatment showed number of electron-dense plaques in the cell membrane and a tendency of decrease in albumin leakage. These results indicate that albumin leakage may be an indirect index of cell wounding in the heart and diltiazem nay be beneficial to protect myocardial cells against isoproterenol-induced cell wounding. It is likely that diltiazem promotes resealing process of the cell membrane.

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저온 스트레스에 발현이 유도되는 콩의 L-asparaginase 유전자의 분리 (Isolation of cDNA Encoding Low Temperature-inducible L-asparaginase from Soybean (Glycin max))

  • 박성환;김기영;진량;이재헌
    • Journal of Plant Biotechnology
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    • 제29권2호
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    • pp.99-104
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    • 2002
  • Suppression subtractive hybridization (SSH)를 통해 저온에 의해 발현이 유도되는 cDNA들을 분리하였고, 그 중 하나인 slti182는 asparaginase 유전자들과 높은 유사성을 보였다. Slti182의 전체 cDNA인 GmASP1은 1258 bp 길이에 326개 아미노산으로 구성된 긴 Open reading frame (ORF)를 포함하고 있었다. GmASP1 단백질은 애기장대의 putative asparaginase (AB012247)와 84%의 유사성을 보였으나, 애기장대의 다른 isoform(Z34884)와는 55%의 유사성을 나타내었다. GmASP1 유전자는 저온 처리 후 3시간째부터 발현이 유도되어 6시간째에 최대발현을 나타내었고, 이후 점차 감소하여 48시간에는 저온처리전 수준으로 되돌아왔다. 또한, 식물체를 저온에서 48시간 둔 후에 다시 상온으로 옮겼을 때, 정상수준으로 떨어진 GmASP1의 발현이 다시 증가하는 양상을 보였다. 이러한 결과로 볼 때, GmASP1이 저온 스트레스에 반응 초기의 단백질 합성 촉진에 중요한 역할을 할 것으로 사료된다.

Isolation of CONSTANS as a TGA4/OBF4 Interacting Protein

  • Song, Young Hun;Song, Na Young;Shin, Su Young;Kim, Hye Jin;Yun, Dae-Jin;Lim, Chae Oh;Lee, Sang Yeol;Kang, Kyu Young;Hong, Jong Chan
    • Molecules and Cells
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    • 제25권4호
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    • pp.559-565
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    • 2008
  • Members of the TGA family of basic domain/leucine zipper transcription factors regulate defense genes through physical interaction with NON-EXPRESSOR OF PR1 (NPR1). Of the seven TGA family members, TGA4/octopine synthase (ocs)-element-binding factor 4 (OBF4) is the least understood. Here we present evidence for a novel function of OBF4 as a regulator of flowering. We identified CONSTANS (CO), a positive regulator of floral induction, as an OBF4-interacting protein, in a yeast two-hybrid library screen. OBF4 interacts with the B-box region of CO. The abundance of OBF4 mRNA cycles with a 24 h rhythm under both long-day (LD) and short-day (SD) conditions, with significantly higher levels during the night than during the day. Electrophoretic mobility shift assays revealed that OBF4 binds to the promoter of the FLOWERING LOCUS T (FT) gene, a direct target of CO. We also found that, like CO and FT, an OBF4:GUS construct was prominently expressed in the vascular tissues of leaf, indicating that OBF4 can regulate FT expression through the formation of a protein complex with CO. Taken together, our results suggest that OBF4 may act as a link between defense responses and flowering.