• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.1499-1506
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• 2016

Background: Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. Materials and Methods: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. Results: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. Conclusions: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.

• 식물병연구
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• 제15권3호
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• pp.254-258
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• 2009

전형적인 모자이크 증상의 표주박(Lagenaria leucantha var. gourda)으로부터 CMV를 분리하고 Lag-CMV로 명명하였다. 표주박에서 분리한 Lag-CMV는 N. benthamiana에서 전신감염되어 전형적인 모자이크 증상을 나타내 대조로 공시한 CMV들과 유사하였다. 그러나 N. tabacum. cv. Xanthi nc에서 접종엽에 퇴록반점이 형성되는 점이 As-CMV와 유사하였으며, 접종엽에 무병징을 나타낸 Fny-CMV와 구분되었다. 또한 Fny-CMV에 감염된 오이와 쥬키니 호박은 매우 심한 모자이크 증상을 나타낸 반면, Lag-CMV와 As-CMV에 감염된 식물은 비교적 엷은 모자이크 병징이 발현되었다. 고추에서는 Fny-CMV와 As-CMV가 접종상엽에 모자이크 증상이 발현되으나, Lag-CMV는 무병징으로 감염되었다. Lag-CMV에 감염된 N. benthamiana로부터 추출한 dsRNA의 전기영동 패턴은 대조의 Fny-CMV나 As-CMV의 dsRNA와 종류 및 분자 크기에서 차이를 나타내지 않았다. Lag-CMV에 감염된 N. benthamiana로부터 추출한 total RNA를 외피단백질 유전자와 IR 및 3' 비번역영역 일부를 포함하는 약 950 bp의 cDNA합성을 위한 프라이머를 이용하여 RT-PCR을 실시한 결과 약 950 bp의 cDNA가 검출되었다. 이 cDNA를 제한효소 EcoRI, HindIII, MspI, SalI 및 XhoI으로 처리한 후 RFLP분석을 실시한 결과, Lag-CMV의 RFLP패턴은 As-CMV와 일치하였다. 한편 이 cDNA를 이용하여 염기서열을 분석한 결과, 염기서열의 유사도는 As-CMV와 99.1%, Fny-CMV와는 94.5%였으며, 외피단백질의 아미노산서열 유사도는 As-CMV는 100%, Fny-CMV와 97.3%를 나타냈다. 이와 같은 결과들로부터 표주박에서 분리한 Lag-CMV는 As-CMV와 같은 서브그룹 IB에 속하는 것으로 확인되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제30권11호
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• pp.1529-1539
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• 2017

Objective: The objective of this study was to compare the DNA methylation profile in the longissimus dorsi muscle between Small Tailed Han and Dorper${\times}$Small Tailed Han crossbred sheep which were known to exhibit significant difference in meat-production. Methods: Six samples (three in each group) were subjected to the methylated DNA immunoprecipitation sequencing (MeDIP-seq) and subsequent bioinformatics analyses to detect differentially methylated regions (DMRs) between the two groups. Results: 23.08 Gb clean data from six samples were generated and 808 DMRs were identified in gene body or their neighboring up/downstream regions. Compared with Small Tailed Han sheep, we observed a tendency toward a global loss of DNA methylation in these DMRs in the crossbred group. Gene ontology enrichment analysis found several gene sets which were hypomethylated in gene-body region, including nucleoside binding, motor activity, phospholipid binding and cell junction. Numerous genes were found to be differentially methylated between the two groups with several genes significantly differentially methylated, including transforming growth factor beta 3 (TGFB3), acyl-CoA synthetase long chain family member 1 (ACSL1), ryanodine receptor 1 (RYR1), acyl-CoA oxidase 2 (ACOX2), peroxisome proliferator activated receptor-gamma2 (PPARG2), netrin 1 (NTN1), ras and rab interactor 2 (RIN2), microtubule associated protein RP/EB family member 1 (MAPRE1), ADAM metallopeptidase with thrombospondin type 1 motif 2 (ADAMTS2), myomesin 1 (MYOM1), zinc finger, DHHC type containing 13 (ZDHHC13), and SH3 and PX domains 2B (SH3PXD2B). The real-time quantitative polymerase chain reaction validation showed that the 12 genes are differentially expressed between the two groups. Conclusion: In the current study, a tendency to a global loss of DNA methylation in these DMRs in the crossbred group was found. Twelve genes, TGFB3, ACSL1, RYR1, ACOX2, PPARG2, NTN1, RIN2, MAPRE1, ADAMTS2, MYOM1, ZDHHC13, and SH3PXD2B, were found to be differentially methylated between the two groups by gene ontology enrichment analysis. There are differences in the expression of 12 genes, of which ACSL1, RIN2, and ADAMTS2 have a negative correlation with methylation levels and the data suggest that DNA methylation levels in DMRs of the 3 genes may have an influence on the expression. These results will serve as a valuable resource for DNA methylation investigations on screening candidate genes which might be related to meat production in sheep.

• Molecular & Cellular Toxicology
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• 제1권1호
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• pp.62-71
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• 2005

For toxicity model in the kidney, renal cell carcinoma (RCC) is one of the most important model to assess the structural and functional alterations. Most RCCs are sporadic, and environmental agents are suspected to play a role in the etiology of the disease. In this study, we discovered novel evidence for previously unknown gene expression patterns related to progression according to cancer stage in RCC. Four clear cell RCC tissue samples along with five corresponding patient-matched normal kidney tissue samples were obtained from patients undergoing partial or radical nephrectomy. To examine the difference of gene expression profile in clear cell RCC, radioactive cDNA microarrays were used to evaluate changes in the expression of 1,152 genes in a total. Using $^{33}P-labeled$ probes, this method provided highly sensitive gene expression profiles including drug metabolism, and cellular signaling. 29 genes were identified with expression levels that differed by more than 2.0 value of z-ratio, compared with that in control. Whereas expression of 38 genes were decreased by less than-2.0 value of z-ratio. In conclusion, this study has identified 67 gene expression alterations in clear-cell type of RCC. Most notably, genes involved in cell growth were up-regulated in stage I more than stage III whereas genes involved in signal transduction were down-regulated in which both stage I and stage III. The identified alteraions of gene expression will likely give in sight in to clear cell RCC and tumor progression.

• 대한수의학회지
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• 제29권3호
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• pp.303-313
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• 1989

To investigate the epidemiological trait of intestinal diseases of animals caused by thermophilic Campyllobacter spp., isolation of etiological agent was carried out and the profiles of plasmids and the transfer of resistance plasmid in the isolated Campylobacter spp. were examined. The results were as follows. 1. A total of 110 isolates of C jejuni and C coli were subjected to the test for the presence of plasmid DNA. Of the isolates examined, 60% of the isolates were noted to harbor plasmid DNA. Plasmid occurrencer ate from pigs, chickens and cattle were 76.2%, 61.7% and 37.7%, respectively. The plasmids of a large molecular weight, ranging from 36 Md to 86Md, were identified with the strains of tetracycline resistant. 2. Transfer frequency of tetracycline resistant plasmids was higher in the case of the filter mating method than in the broth mating method by the factor of 10~1,000. 3. Tetracycline resistant plasmids of C jejuni were transferrable to C jejuni and C coli by conjugation. In a low frequency, the transfer of tetracycline plasmid was also possible to Vibrio parahemolyticus. However, it was impossible to transfer to Streptococcus fecalis, E coli and Vibrio cholerae. 4. Tetracycline resistant plasmids of C jejuni were impossible to transfer to Campylobacter spp. and related bacteria by transformation.

• Journal of Acupuncture Research
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• 제24권1호
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• pp.151-163
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• 2007

Objectives : Juglandis Semen herbal acupuncture solution(JSS) has a broad array of clinical applications in oriental medicine, including treatment of chronic musculoskeletal diseases such as arthritis. This study was performed to investigate the global gene expression profiles using microarray assay in RAW 264.7 cell line treated with JSS and to advance our understanding of the pharmacologic effect of JSS. Methods : Change of the gene expression profile in RAW cell line following treatment with lipopolysaccharide(LPS) alone, or with LPS plus JSS was investigated with a cut-off level of 2 fold change in the expression. Especially, Change of the gene expression by treatment with LPS alone was compared with that by treatment with LPS plus JSS with a cut-off level of 1/2 fold change in the expression. Results: Of the 8170 genes profiled in this study, 51 were upragulated and 21 downregulated following LPS treatment, and 88 were upregulated and 69 downregulated following costimulation of JSS and LPS. Of the 51 genes upregulated following LPS treatment, 10 were downregulated following costimulation of JSS and LPS. Of the 21 genes downregulated following LPS treatment, 3 were upregulated following costimulation of JSS and LPS. Conclusion : JSS treatment induced upregulation of some genes including IL-10 and downregulation of that including MMP13 with its possible implication in an antiinflammatory action of JSS. However, further research on expression profile changes induced by JSS treatment is expected.

• 대한의용생체공학회:학술대회논문집
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• 대한의용생체공학회 1998년도 추계학술대회
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• pp.66-69
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• 1998

Comparative Genomic Hybridization (CGH)은 세포 내 특정 DNA 서열 이상을 염색체상에 보여주는 중요한 분자 세포 유전학 기법이다. CGH 기법에서는 세포 분열 중기의 염색체에서 준비한 형광 비율 영상의 정량적 분석을 위해서 Digital 영상 처리 기술이 쓰여야 한다. 본 논문에서는 최근 연구 개발된 영상 처리 algorithm들이 어떻게 CGH 기법에 쓰이는 지를 소개하려 한다. 각 염색체의 형광 비율 profile를 평균하기 위해, 염색체 영상의 이원화, 염색체 영상 뼈대 변환(skeletonization), 뼈대 정보의 변수화와 영상 명암의 재추출을 통한 굽은 염색체 영상 펴기 등이 언구되었다. 개발된 algorithm 들은 바이오메드랩 사의 ChIPS 핵형 정렬 시스템에 구현했다.

• 한국수산과학회지
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• 제48권3호
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• pp.346-353
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• 2015

Glucocorticoids (GCs) are steroid hormones regulated through responses to stress to maintain diverse metabolic and homeostatic functions. GCs act on the glucocorticoid receptor (GR), a member of the nuclear receptor family. This study identified and characterized the GR gene from the big-belly seahorse Hippocampus abdominalis designating it HaGR. The open reading frame of the HaGR cDNA was 2,346 bp in length, encoding a 782-amino-acid polypeptide with a theoretical isoelectric point of 6.26 and predicted molecular mass of 86.8 kDa. Nuclear receptors share a common structural organization, comprising an N-terminal transactivation domain, DNA-binding domain, and C-terminal ligand-binding domain. The tissue-specific mRNA expression profile of HaGR was analyzed in healthy seahorses using a qPCR technique. HaGR mRNA was expressed ubiquitously in all of the tissues examined, with the highest expression levels in kidney, intestine, stomach, and gill tissues. The mRNA expression in response to immune challenge with lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), Edwardsiella tarda, and Streptococcus iniae revealed that it is inducible in response to pathogen infection. These results suggest that HaGR is involved in the immune response of the big-belly seahorse.

• 한국균학회지
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• 제39권3호
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• pp.180-184
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• 2011

본 연구는 URP-PCR 핵산지문법을 이용한 Paecilomyces spp.와 Cordyceps spp.의 종간, 종내 유전적 다양성분석을 실시 하였다. 12종류의 20mer의 URP primer가적용된 바 URP2F, URP2R, URP9F, URP4R, URP17R는 종간에 특이적인 PCR다형성밴드를 형성하였으며 Cordypces militaris 균주간에는 4가지 type의 PCR 다형성이 관찰되었다. 그러나 Paecilomyces japonica의 균주내에는 동일한 PCR 다형성을 나타내었다. URP-PCR profile을 이용하여 경동시장에서 수집한 미 동정 동충하초를 동정 할 수 있었다.

• Journal of Plant Biotechnology
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• 제35권2호
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• pp.133-140
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• 2008

DNA와 histone 단백질의 변형은 식물 발달에 상당히 중요한 역할을 하는 것으로 알려져 있다. 식물 조직 배양 및 식물 발달 단계에서 methylation의 영향을 알아보고자 벼 종자로부터 캘러스 형성 및 식물체 재분화 단계에서 demethylation 물질인 5-azacytidine을 처리하여 유전자 발현 양상을 분석하였다. 식물체로의 재분화 능력이 있는 벼 배상체 캘러스는 5-azaC가 첨가된 H6A 배지에서는 형성되지 않았으며 갈색을 띠는 캘러스가 형성되었다. 또한 정상적인 캘러스를 5-azaC가 첨가된 MSRA 재분화 배지에서 배양했을 때도 대조구와는 달리 식물체 재분화는 이루어지지 않았다. 이러한 결과는 5-azaC가 정상적인 배상체 캘러스 및 shoot 분화에 부정적인 영향을 미친다는 것을 나타냈으며 따라서 DNA methylation이 식물 조직배양에서의 정상적인 세포 dedifferentiation과 differentiation에 필수 요인이라는 것을 알 수 있었다. 벼 캘러스 형성 및 재분화 과정 동안의 methylation 영향을 알아보고자 각 단계별로 5-azaC를 처리 후 $GeneFishig^{TM}$ DEG와 DNA chip을 사용하여 유전자 발현 양상을 분석하였다. Epigenetic regulation, 전자전달, 핵산대사, 스트레스 반응에 관여하는 일부 유전자들의 발현이 증가하거나 감소하는 것을 알 수 있었다. 발현 차이가 있는 일부 유전자를 클로닝하여 확인하였고 RT-PCR 및 northern 분석으로 각 단계에서의 발현 차이를 할인하였다.