• Title/Summary/Keyword: conventional-PCR

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Expression Levels of Tetraspanin KAI1/CD82 in Breast Cancers in North Indian Females

  • Singh, Richa;Bhatt, Madan Lal Brahma;Singh, Saurabh Pratap;Kumar, Vijay;Goel, Madhu Mati;Mishra, Durga Prasad;Srivastava, Kirti;Kumar, Rajendra
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3431-3436
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    • 2016
  • Background: Carcinogenesis is a multifaceted intricate cellular mechanism of transformation of the normal functions of a cell into neoplastic alterations. Metastasis may result in failure of conventional treatment and death Hence, research on metastatic suppressors in cancer is a high priority. The metastatic suppressor gene CD82, also known as KAI1, is a member of the transmembrane 4 superfamily which was first identified in carcinoma of prostate. Little work has been done on this gene in breast cancer. Herein, we aimed to determine the gene and protein level expression of CD82/KAI1 in breast cancer and its role as a prognosticator. Materials and Methods: In this study, 83 histologically proven cases of breast cancer and a similar number of controls were included. Patient age ranged from 18-70 years. Quantitative Real Time Polymerase Chain Reaction (q-RT PCR) and immunohistochemistry (IHC) were used to investigate KAI1 expression at gene and protein levels, respectively. Statistical analysis was done to correlate expression of KAI1 and clinicopathological parameters. Results: It was revealed that: (i) KAI1 was remarkably diminished in metastatic vs non metastatic breast cancer both at the gene and the protein levels (P < .05); (ii) KAI1 expression levels were strongly correlated with TNM staging, histological grade and advanced stage (p<0.001) and no association was found with any other studied parameter; (iii) Lastly, a significant correlation was observed between expression of KAI1 and overall median survival of BC patients (P = 0.04). Conclusions: Our results suggest that lack of expression of the KAI1 might indicate a more aggressive form of breast cancer. Loss of KAI1 may be considered a significant prognostic marker in predicting metastatic manifestation. When evaluated along with the clinical and pathological factors, KAI1 expression may be beneficial to tailor aggressive therapeutic strategies for such patients.

Construction and Production of Concatameric Human TNF Receptor-Immunoglobulin Fusion Proteins

  • Yim, Su-Bin;Chung, Yong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.81-89
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    • 2004
  • Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and lymphotoxin-$\alpha$ (LT-$\alpha$, TNF-$\beta$) can initiate and perpetuate human diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA), and insulin-dependent diabetes mellitus (IDDM). TNFs can be blocked by the use of soluble TNF receptors. However, since monomeric soluble receptors generally exhibit low affinity or function as agonists, the use of monomeric soluble receptors has been limited in the case of cytokines such as TNF-$\alpha$, TNF-$\alpha$, interleukin (IL)-1, IL-4, IL-6, and IL-13, which have adapted to a multi component receptor system. For these reasons, very high-affinity inhibitors were created for the purpose of a TNFs antagonist to bind the TNFR and trigger cellular signal by using the multistep polymerase chain reaction method. First, recombinant simple TNFR-Ig fusion proteins were constructed from the cDNA sequences encoding the extracellular domain of the human p55 TNFR (CD120a) and the human p75 TNFR (CD120b), which were linked to hinge and constant regions of human $IgG_1$ heavy chain, respectively using complementary primers (CP) encoding the complementary sequences. Then, concatameric TNFR-Ig fusion proteins were constructed using recombinant PCR and a complementary primer base of recombinant simple TNFR-Ig fusion proteins. For high level expression of recombinant fusion proteins, Chinese hamster ovary (CHO) cells were used with a retroviral expression system. The transfected cells produced the simple concatameric TNFR-Ig fusion proteins capable of binding TNF and inactivating it. These soluble versions of simple concantameric TNFR-Ig fusion proteins gave rise to multiple forms such as simple dimers and concatameric homodimers. Simple TNFR-1g fusion proteins were shown to have much more reduced TNF inhibitory activity than concatameric TNFR-Ig fusion proteins. Concatameric TNFR-Ig fusion proteins showed higher affinity than simple TNFR-Ig fusion proteins in a receptor inhibitor binding assay (RIBA). Additionally, concatameric TNFR-Ig fusion proteins were shown to have a progressive effect as a TNF inhibitor compared to the simple TNFR-Ig fusion proteins and conventional TNFR-Fc in cytotoxicity assays, and showed the same results for collagen induced arthritis (CIA) in mice in vivo.

The Chemotoxonoic Relationship of Vibrio cholerae non-O1 by Fatty Acid Compositions (지방산 조성에 의한 vibrio cholerae non-O1의 화학분류학적 관계)

  • 성희경;이원재;장동성
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.149-154
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    • 1998
  • The authors attempted utilization of fatty acid composition of vibrios as a tool for identification of the strains. Fatty acid of 49 strains of Vibrio cholerae non-O1, V. cholerae O1, V mimicus, V vulinificus and V parahaemolyticus was analyzed by gas-liquid chromatography column. According to the statistical analysis of the fatty acid data, the relationship between the Vibrio species and serotypes of the strains was discussed. Forty one kinds of fatty acid were detected from the tested strains and 35 kinds of fatty acids among the detected fatty acids were significant factors to identify the vibrios. The predominant fatty acids were 16:0, 16:1 cis 9, 18:1 trans 9/6/cis 11 and 15:0 iso 2OH/16:1 cis 9 as above about 20% in total. Fatty acid compositions of the Vibrio species were an important factor in identifying their subspecies either predominant fatty acids or minor ones. According to the analysed results by a conventional statistical processing method (UPGMA) and prepared dendrogram, V cholerae non-01 had more closer relationship with V. mimicus compared with V. cholerae 01. Moreover, the distribution of hydroxy acid was a significant factor for identifying V cholerae subspecies. Comprising all the 10 serotypes detected from V. cholerae non-01 examined such as O2, O5, O8, O10, O14, O27, O37, O39, O45 and O69, we could group them into seven subspecies by cluster analysis with the similarity value of fatty acid composition as above 92%. It means that there is a significant relationship between serotypes and fatty acid composition of V. cholerae. These results indicated that numerical analysis of fatty acid composition data of V cholerae non-01 could classifY them into subspecies, and also which may provide a useful epidemiologic information or a basis for further analysis such as PCR and DNA probe analysis.

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A Study on the Sensory Characteristic of Yogurt and Antimicrobial Activity of Lactobacillus plantarum LHC52 Isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum LHC52의 항균활성과 요구르트의 관능성 연구)

  • Lee, Seung-Gyu;Han, Ki-Sung;Jeong, Seok-Geun;Oh, Mi-Hwa;Jang, Ae-Ra;Kim, Dong-Hun;Bae, In-Hyu;Ham, Jun-Sang
    • Food Science of Animal Resources
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    • v.30 no.2
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    • pp.328-335
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    • 2010
  • The aim of our study was to develop a new starter culture for fermented milk. Polymerase chain reaction screening of 103 acid-producing isolates from Kimchi identified 72 Lactobacillus strains. The ability of the strains to inhibit the growth of the food-borne human pathogens (Escherichia coli, Salmonella Enteritidis, Staphylococcus aureus) was measured, using a conventional paper disk method. Among the 72 strains, strain LHC52 displayed potent antagonistic activity. Use of 16S rDNA sequencing and the API 50CHL system identified the strain as Lactobacillus plantarum and it was designated L. plantarum LHC52. Biochemical analyses revealed especially high antibacterial activity against E. coli. Yogurt produced using L. plantarum LHC52 did not show different microbiological and physicochemical properties compared to conventionally-prepared yogurt, implicating L. plantarum LHC52 as a useful, potently antibacterial starter culture for yogurt preparation.

Diagnostic Availability of Estrogen Receptor Alpha mRNA on Cervical Cancer Tissue (자궁경부암 조직에서 에스트로겐 수용체 알파 mRNA의 진단적 유용성)

  • Kim, Geehyuk;Yu, Kwangmin;Kim, Jungho;Kim, Seoyong;Park, Sunyoung;Ahn, Sungwoo;Lee, Ji-Young;Kim, Sunghyun;Park, Ho-Hyun;Lee, Dongsup
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.449-456
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    • 2018
  • Cervical cancer is the fourth most frequently diagnosed cancer in women worldwide. In lower Human Development Index countries, it has the second highest incidence and mortality among cancer in women. Therefore, better diagnosis and treatment systems are needed. Among them, estrogen receptor alpha ($ER-{\alpha}$) mRNA expression has been analyzed with RT-qPCR since several studies reported that $ER-{\alpha}$ is necessary in the maturation of the uterus and is related to cervical cancer. In this study, $ER-{\alpha}$ quantitative analysis was performed on various lesions and normal tissue samples. Based on the receiver operating characteristic (ROC) curve, its sensitivity and specificity were 85% and 75%, respectively, showing higher or similar results to those of conventional HPV tests. In addition, its expression level was analyzed with clinical information. With regression analysis, the R square value between the $ER-{\alpha}$ mRNA expression level and menopause status was 0.5041, indicating a strong correlation. This study was performed as part of a pilot study and suggests that $ER-{\alpha}$ is related to carcinogenesis. Future studies will examine other hormones and menopausal factors with a larger sample size.

Determination of Fire Blight Susceptibility on Wild Rosaceae Plants in Korea by Artificial Inoculation (인공접종을 통한 국내 야생 장미과 식물의 화상병 감수성 검정)

  • In Woong Park;Yu-Rim Song;Eom-Ji Oh;Yoel Kim;In Sun Hwang;Mi-Jin Jeon;Chorong Ahn;Jin-Suk Kim;Soonok Kim;Chang-Sik Oh
    • Research in Plant Disease
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    • v.29 no.1
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    • pp.23-38
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    • 2023
  • The fire blight caused by Erwinia amylovora (Ea) is a devastating disease of Rosaceae plants, including commercially important apple and pear trees. Since the first report in Korea in May 2015, it has been spreading to neighboring regions gradually. Host plants can be infected by pollinators like bees, rainfall accompanied by wind, and cultural practices such as pruning. Many studies have revealed that wild Rosaceae plants such as Cotoneaster spp., Crataegus spp., Pyracantha spp., Prunus spp., and Sorbus spp. can be reservoirs of Ea in nature. However, wild Rosaceae plants in Korea have not been examined yet whether they are susceptible to fire blight. Therefore, the susceptibility to fire blight was examined with 25 species in 10 genera of wild Rosaceae plants, which were collected during 2020-2022, by artificial inoculation. Bacterial suspension (108 cfu/ml) of Ea type strain TS3128 was inoculated artificially in flowers, leaves, stems, and fruits of each plant species, and development of disease symptoms were monitored. Moreover, the presence of Ea bacteria from inoculated samples were checked by conventional polymerase chain reaction. Total 14 species of wild Rosaceae plants showed disease symptoms of fire blight, and Ea bacteria were detected inside of inoculated plant parts. These results suggest that wild Rosaceae plants growing nearby commercial apple and pear orchards in Korea can be Ea reservoirs, and thus they should be monitored regularly to minimize the damage by Ea infection and spreading.