• Title/Summary/Keyword: HC2 assay

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Differential Regulation of the Caprine ${\beta}$-Lactoglobulin Gene Promoter in the Cultured Mammary HC11 Cells

  • Kim, Jae-Man
    • Animal cells and systems
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    • v.1 no.2
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    • pp.345-350
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    • 1997
  • The ${\beta}$-Lactoglobulin (BLG) gene expression is differentially regulated during development of the mammary tissues. Such differential regulation of the BLG gene expression can be reiterated in the cultured mammary HC11 cells. In the growing non-confluent HC11 cells, the BLG promoter activity was shown to be partially repressed by the upstream regulatory sequence. The repression was gradually diminished and switched to activation as the cells grew confluent. The differential regulation of the BLG promoter was controlled by the 5'-regulatory sequence located at the upstream of 205 bp. Electromobility shift assay showed that nuclear extract from HC11 cells differentially bound on the regulatory sequence, depending on the cell confluency, which was in accordance with the differential transcriptional activity. DNase I foot-print assay, however, revealed that all nuclear extracts presented the same foot-prints, regardless of confluency of HC11 cells. These results suggest that differential regulation BLG gene expression by the 5'-regulatory sequence may be accomplished by competitive and/or cooperative binding of differential regulatory factors on the same regulatory element.

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Hybrid Capture 2 Assay Based Evaluation of High-Risk HPV Status in Healthy Women of North-East India

  • Rai, Avdhesh Kumar;Das, Debabrata;Kataki, Amal Chandra;Barmon, Debabrata;Aggarwal, Dippy;Deka, Pankaj;Shrivastava, Sushruta;Sharma, Jagannath Dev;Sarma, Anupam;Baruah, Upasana;Sharma, Mousumi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.861-865
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    • 2014
  • Background: High risk HPV (HR-HPV) testing has been recommended as an effective tool along with cytology screening in identification of cervical intraepithelial lesions (CINs) and prevention of their progress towards invasive cervical cancer. The aim of this study was to assess the HR-HPV DNA status by Hybrid Capture 2 (HC2) assay in healthy asymptomatic women of North-East India. Materials and Methods: This study examined cervical cell samples of forty three (n=43) healthy women by HC2 assay. A High Risk HPV DNA kit (Qiagen) was used which can detect 13 high risk HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68. Results: The mean relative light units (RLU) for samples was in the range of 141-5, 94, 619. HR-HPV DNA was confirmed in 16% (7/43) of participant women samples. Among demographic and clinical parameters, menstrual irregularity (p=0.039) and infection history (p=0.028) has shown statistically significant differences between the HR-HPV-positive and negative groups. In the HR-HPV positive group, two women were confirmed for CINs after colposcopy and histopathologic examination. Conclusions: We suggest that there may be an association between irregular menstruation and infection history of the urogenital tract with HR-HPV DNA prevalence in North-East Indian asymptomatic women. HC2 assay can be a valuable tool for HR-HPV screening.

Construction of a Baculovirus Expression System Using Hyphantria cunea Nuclear Polyhedrosis Virus for Eukaryotic Cells

  • Lee, Hyung-Hoan;Kang, Bong-Joo;Park, Kap-Ju;Cha, Soung-Chul
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.676-684
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    • 1998
  • Baculovirus transfer and expression vectors with Hyphantria cunea nuclear polyhedrosis virus (HcNPV) were constructed. An initial transfer vector, pHcEV, constructed using HcNPV was previously reported (Park et al. 1993. J. Kor. Soc. Viral. 23: 141-151). Herein, the size of the vector was properly reduced, and a functionally perfect vector was constructed and named pHcEV-IV (6.7 kb). The vector has a 2.2-kb HcNPV DNA sequence in the 5'-flanking region of the vector's polyhedrin gene promoter. The 1.8-kb HcNPV DNA sequence, poly A signal sequence, T3 primer sequence, and 13 multicloning site sequences, in order, were ligated in front of the translation start codon of the polyhedrin gene. The cloning indicating marker lacZ gene was inserted into the pHcEV-IV, named pHcEV-IV-lacZ, and transferred into the wild-type virus. Recombinant expression virus, lacZ-HcNPV, was constructed by replacing the lacZ gene in the pHcEV-IV-lacZ with the polyhedrin gene of the wild-type virus. The recombinant virus was isolated from blue plaques that produce $\beta$-galactosidase without polyhedra. The lacZ gene insertion was confirmed by Southern hybridization analysis. The expression of the lacZ gene in Spodoptera frugiperda cells infected with the lacZ-HcNPV was examined by SDS-PAGE and colorimetric assay. One 116-kDa LacZ protein band appeared on the PAGE. The production rate of the $\beta$-galactosidase was approximately 50 international units (IU) per min per ml between 2 to 5 days postinfection (p.i.). The highest activity occurred at five days p.i. was 170 IU/min/$m\ell$. The enzyme activity first appeared about 20 h p.i. as measured by colorimetric assay.

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Comparison of Clinical Efficacy between an HPV DNA Chip and a Hybrid-Capture II Assay in a Patient with Abnormal Colposcopic Findings (질 확대경상 비정상 소견을 보인 환자에서 HPV DNA chip과 Hybrid-Capture II assay의 임상적 유용성 비교)

  • Kim, Tae-Jung;Jung, Chan-Kwon;Lee, Ah-Won;Jung, Eun-Sun;Choi, Young-Jin;Lee, Kyo-Young;Park, Jong-Sup
    • The Korean Journal of Cytopathology
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    • v.19 no.2
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    • pp.119-125
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    • 2008
  • This study was performed to compare the efficacy between a DNA chip method and a Hybrid-Capture II assay (HC-II) for detecting human papillomavirus in patients with intraepithelial lesions of the uterine cervix. From May, 2005, to June, 2006, 192 patients with abnormal colposcopic findings received cervical cytology, HC-II and HPV DNA chip tests, and colposcopic biopsy or conization. We compared the results of HC-II and HPV DNA chip in conjunction with liquid based cervical cytology (LBCC) and confirmed the results of biopsy or conization. The sensitivity of the HPV DNA chip test was higher than HC-II or LBCC. The HPV DNA chip in conjunction with LBCC showed higher sensitivity than any single method and higher sensitivity than HC-II with LBCC. We confirmed that the HPV DNA chip test was more sensitive for detecting HPV in cervical lesions than HC-II, and that it would provide more useful clinical information about HPV type and its multiple infections.

Effects of Herbar Chelidonii on the Cultured Spinal Sensory Neurons Damaged by XO/HX (백굴채(白屈菜)가 손상된 배양척수감각신경세포에 미치는 영향)

  • Shin, Byung-Cheul;Song, Yung-Sun
    • The Journal of Korea CHUNA Manual Medicine
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    • v.2 no.1
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    • pp.143-157
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    • 2001
  • Objectives and Methods : To evaluate the mechanism of oxidative damage by xanthine oxydase(XO) and hypoxanthine(HX)-induced oxygen radicals, MTT assay and NR assay were carried out after the cultured mouse spinal sensory neurons were preincubated for 4 hours with various concentrations of XO/HX. And the amount of total protein. neurofilament EIA. lipid peroxidation and LDH activity were measured, to evaluate the protective effect of Herbar Chelidonii(HC) water extract on cultured spinal sensory neurons damaged by XO/HX. after the cultured mouse spinal sensory neurons were preincubated with various concentrations of HC water extract for 3 hours prior to exposure of XO/HX. Results : XO/HX decreased significantly the survival rate of the cultured mouse sensory neurons by NR assay and MTT assay In proportion to concentration and exposed time. In proportion to concentration and exposed time on cultured spinal sensory neurons, XO/HX showed the quantitative decrease of neurofilament by EIA. the decrease of total protein amount by SRB assay and the Increase of lipid peroxidation as well as LDH. HC showed the quantitative increase of neurofilament and total protein, but showed the decrease of lipid peroxidation and LDH activity against the neurotoxicity of XO/HX. Conclusions : From the above results, it is concluded that XO/HX have a neurotoxic effect on cultured spinal sensory neurons and that the herbs extract, such as HC, prevent the toxicity of XO/HX effectively in that they decrease lipid peroxidation and LDH activity.

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Biocontrol of gray mold of cucumber and tomato by epihpytic bacteria in field condition

  • Lee, Sang-Yeob;Lee, Sang-Bum;Kim, Il-Yon;Kim, Yong-Ki
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.104.2-105
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    • 2003
  • Bacterial isolates, CC178, PTC25, HC39 and KY165 originally obtained from the leaves of cucumber or tomato were selected for biocontrol agents against gray mold of cucumber and tomato by in vivo cucumber seedling assay. Each suspension of the selected epiphytic bacteria were sprayed three times at seven-day interval from early stage of cucumber in a field. Incidence of gray mold on cucumber fruits treated with isolates CC178, PTC25, HC39 and KY165 was 15.3%, 18.2%, 23.6%, and 10.4%, respectively, whereas that of control was 38.0% after 7 days of final spray. On the other hand, treatment with the selected isolates, CC178, PTC25, HC39, and KY165 on tomato showed 2.2%, 1.3%, 2.9%, and 3.5% in the incidence of gray mold on leaves, whereas that of control was 9.3%. All selected isolates had strong antagonistic activity against Botrytis cinerea on dual culture plate assay.

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Effects of Chungkukjang Added with Onion on Lipid and Antioxidant Metabolisms in Rats Fed High Fat-Cholesterol Diet (양파 첨가 청국장이 고콜레스테롤혈증 유발 흰쥐의 지질 및 항산화 대사에 미치는 영향)

  • Park, Jae-Hee;Kim, Jung-Mi;Park, Eun-Ju;Lee, Kyung-Hea
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.10
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    • pp.1244-1250
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    • 2008
  • This study was carried out investigate the effects of chungkukjang added with onion (OC) and chungkukjang (C) on the lipid and antioxidant metabolisms of hypercholesterolemic rats. Thirty-two male Sprague-Dawley rats were divided into four groups after 1-week adaptation period and were fed a normal diet, high fat-cholesterol diet (HC), HC-OC or HC-C for 8 weeks, respectively. The supplementation of HC-OC and HC-C groups significantly reduced hepatic total cholesterol and AST activity. HC-C group increased high density lipoprotein, while decreasing low density lipoprotein and AI compared with HC-OC group. Conjugated dienes of HC-C group was significantly lower than those of HC-OC group. $H_2O_2$ induced DNA damage reduced significantly in HC-OC and HC-C groups compared with high fat-cholesterol diet group. $H_2O_2$ induced DNA damage exhibited significant positive correlations with hepatic total cholesterol, AST and CD. These results suggested that supplementation of chungkukjang or chungkukjang added with onion might be helpful in preventing lipid oxidation and leukocytic DNA damage. However, the health beneficial effect has not improved by the addtion of onion in chungkukjang.

PAX1 Methylation Analysis by MS-HRM is Useful in Triage of High-grade Squamous Intraepithelial Lesions

  • Wang, Zhen-Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.891-894
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    • 2014
  • This study is aimed to investigate the role of paired boxed gene 1 (PAX1) methylation analysis by methylation-sensitive high-resolution melting (MS-HRM) in the detection of high grade lesions in atypical squamous cells cannot exclude high-grade squamous intraepithelial lesion (ASC-H) and compared its performance with the Hybrid Capture 2 (HC2) human papillomavirus (HPV) test. In our study, 130 cases with a diagnosis of ASC-H from the cervical cytological screening by Thinprep cytologic test (TCT) technique were selected for triage. Their cervical scrapings were collected and evaluated by using PAX1 methylation analysis (MS-HRM) and high-risk HPV DNA test (HC2), followed by colposcopy and cervical biopsy. Chi-square test were used to test the differences of PAX1 methylation or HPV infection between groups. In the detection of CIN2+, the sensitivity, specificity, the PPV, NPV and the accuracy of PAX1 MS-HRM assay and high-risk HPV (HR-HPV) tests were respectively 80.6% vs 67.7%, 94.9% vs 54.5%, 83.3%, vs 31.8%, 94.0% vs 84.4%, and 91.5% vs 57.7%. The PAX1 MS-HRM assay proved superior to HR-HPV testing in the detection of high grade lesions (CIN2+) in ASC-H. This approach could screen out the majority of high grade lesion cases of ASC-H, and thus could reduce the referral rate to colposcopy.

Cholesterol Lowering Effect of Ethanol Extracts of Salted and Fermented Small Shrimp in Rats Administered a High Fat Diet (발효 새우젓의 에탄올 추출물이 고지방 식이를 급여한 흰쥐의 콜레스테롤 저하 효과)

  • Kim, Yong-Hyun;Han, Kook-Il;Jeon, Mi-Ae;Kwon, Hyun-Jung;Park, Min-Kyung;Han, Man-Deuk
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.281-286
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    • 2011
  • This study was performed to investigate the effects of salted and fermented shrimp ethanol extract (SFS) on serum lipid metabolism and hepatocytes in rats. Male Sprague-Dawley rats were administered 60% fat feed to induce hypercholesterolemia and were divided into five groups. Experimental groups were classified according to administered diet: normal diet group (NC), high cholesterol diet group (HC), high cholesterol and low dose shrimp extract (20 mg/kg) group (HC-SFSL), high cholesterol and high dose shrimp extract (200 mg/kg) group (HC-SFSH), and high cholesterol and lovastatin (20 mg/kg) group (HC-Lov). The experimental diets were fed ad libitum for 14 days. Compared with the control group, the serum cholesterol and triglycerides were 40.4 and 64.7% lower in the group fed HC-SFSH respectively. Low density lipoprotein (LDL)-cholesterol concentration in serum decreased in the HC-SFSH group compared with the HC group. In a histological assay, hepatocytes in the HC group showed that the vacuolated cells by fat appear clear due to the large amount of intracytoplasmic fat, whereas the liver hepatocytes in the group fed SFS effectively decreased fatty liver and intracytoplasmic fats. These results suggest that the extract of salted and fermented shrimp has an antiatherosclerotic effect and may lessen the effects of cardiovascular disease by reducing the cholesterol level in serum.

Effects of cholesterol and Lactobacillus acidophilus on testicular function

  • Ciftci, Gulay;Tuna, Elif
    • Clinical and Experimental Reproductive Medicine
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    • v.48 no.3
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    • pp.229-235
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    • 2021
  • Objective: In this study, the effects of Lactobacillus acidophilus on testosterone (TES), follicle-stimulating hormone (FSH), luteinizing hormone (LH), androgen-binding protein (ABP), factor-associated apoptosis (FAS), and total cholesterol (TC), as well as histopathological changes, were investigated in male rats fed a high-cholesterol diet. Methods: The study included three groups. The control (C) group was fed standard-diet for 8 weeks. The hypercholesterolemia (HC) group was fed a 2% cholesterol-diet for 8 weeks. The therapeutic group (HCL) was fed a 2% cholesterol-diet for 8 weeks and administered L. acidophilus for the last 4 weeks. FSH, TES, and FAS levels in testicular tissue were determined using an enzyme-linked immunosorbent assay (ELISA), while another sample was examined histopathologically. LH and ABP levels were determined using ELISA, and serum TC levels were assessed via an autoanalyzer. Results: In the HC group, the TC levels were significantly higher and the LH levels were lower (p<0.05) than in the C group. The ABP levels were lower (p>0.05). In the HCL group, the LH and ABP levels were higher (p>0.05) and the TC level significantly lower (p<0.05) than in the HC group. The TES and FSH levels were lower, and the FAS levels were higher, in the HC than in the C group (p<0.05). In the HCL group, levels of all three resembled control levels. Histologically, in the testicular tissue of the HC group, the cells in the tubular wall exhibited atrophy, vacuolization, and reduced wall structure integrity. However, in the HCL group, these deteriorations were largely reversed. Conclusion: Supplementary dietary administration of an L. acidophilus to hypercholesterolemic male rats positively impacted testicular tissue and male fertility hormone levels.