• Title/Summary/Keyword: Genetic screening

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Lack of Association between CYP1A1 M2 and M4 Polymorphisms and Breast Carcinoma in Jordanian Women: a Case-Control Study

  • Amrani, Iman;Bulatova, Nailya;Awidi, Abdalla;Yousef, Al-Motassem;Melhem, Jamal Masad;Al-Masri, Mahmoud;Tahoun, Laila Abu
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.387-393
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    • 2016
  • Background: CYP1A1 is a candidate gene for low-penetrance breast cancer susceptibility, as it plays an important role in the metabolism of carcinogens and estrogens. Purpose: The objective of this study was to assess the association between M2 (A2455G, Ile462Val) and M4 (C2453A, Thr461Asn) polymorphisms in CYP1A1 and breast cancer risk among Jordanian women and in subgroups stratified by menopausal status and smoking history. Materials and Methods: Blood samples were collected from 112 breast cancer female patients and 115 age-matched controls who underwent breast cancer screening with imaging and showed negative results (BI-RADS I or BI-RADS II). Genotyping was performed using the PCR-RFLP technique. Results: No statistically significant overall association was found between breast cancer risk and CYP1A1 M2 genotypes (p= 0.55; OR = 0.77; 95% CI= 0.32 - 1.83) nor with the M4 polymorphism (p= 0.95; OR= 0.95; 95% CI= 0.51 - 1.88). Analysis of subgroups defined by menopausal status or smoking history also revealed no association with these polymorphisms. Furthermore, the four identified haplotypes (AC; AA; GC and GA) were equally distributed among cases and controls, and haplotype analysis showed a strong linkage disequilibrium of both studied loci in either cases or controls (D'=1). Conclusions: Based on the study results, CYP1A1 M2 and M4 polymorphisms do not seem to play a major role in breast cancer risk among Jordanian females.

Comparative Serum Proteomic Analysis of Serum Diagnosis Proteins of Colorectal Cancer Based on Magnetic Bead Separation and MALDI-TOF Mass Spectrometry

  • Deng, Bao-Guo;Yao, Jin-Hua;Liu, Qing-Yin;Feng, Xian-Jun;Liu, Dong;Zhao, Li;Tu, Bin;Yang, Fan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6069-6075
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    • 2013
  • Background: At present, the diagnosis of colorectal cancer (CRC) requires a colorectal biopsy which is an invasive procedure. We undertook this pilot study to develop an alternative method and potential new biomarkers for diagnosis, and validated a set of well-integrated tools called ClinProt to investigate the serum peptidome in CRC patients. Methods: Fasting blood samples from 67 patients diagnosed with CRC by histological diagnosis, 55 patients diagnosed with colorectal adenoma by biopsy, and 65 healthy volunteers were collected. Division was into a model construction group and an external validation group randomly. The present work focused on serum proteomic analysis of model construction group by ClinProt Kit combined with mass spectrometry. This approach allowed construction of a peptide pattern able to differentiate the studied populations. An external validation group was used to verify the diagnostic capability of the peptidome pattern blindly. An immunoassay method was used to determine serum CEA of CRC and controls. Results: The results showed 59 differential peptide peaks in CRC, colorectal adenoma and health volunteers. A genetic algorithm was used to set up the classification models. Four of the identified peaks at m/z 797, 810, 4078 and 5343 were used to construct peptidome patterns, achieving an accuracy of 100% (> CEA, P<0.05). Furthermore, the peptidome patterns could differentiate the validation group with high accuracy close to 100%. Conclusions: Our results showed that proteomic analysis of serum with MALDI-TOF MS is a fast and reproducible approach, which may provide a novel approach to screening for CRC.

Survival of Patients with Prostate Cancer in Yazd, Iran

  • Zahir, Shokouh Taghipour;Nazemian, Mohammad Reza;Zand, Sanaz;Zare, Samad
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.883-886
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    • 2014
  • Background: Prostate cancer is the second leading cause of cancer death in men worldwide. Several factors such as availability of screening tests, and dietary, other lifestyle, environmental and genetic influences contribute to worldwide disparities in prostate cancer incidence and mortality rates. Our aims were to investigate patient characteristics at the time of diagnosis, common treatment strategies employed and survival in an Iranian male population with prostate cancer. Materials and Methods: Archives of Pathology Departments of five referral centers affiliated with the School of Medicine of Shahid Sadoughi University in Yazd province were reviewed. Paraffin-embedded blocks were reviewed by two independent pathologists to confirm the diagnosis. The latest modification of the Gleason Scoring System was adopted to determine pathological grading. Following pathological evaluation, patients were contacted via telephone to acquire information regarding their current status. Results: Pathology blocks were available for 113 patients. However, upon phone contacts, we were unable to determine the survival status in 23 patients (response rate=83%). Therefore, 90 patients were enrolled in the final analysis. The median follow-up time was 6.0 years (ranging from 0.3 to 8.8 years). There were 30 death attributed to prostate cancer in the study group. Kaplan-Meier analysis revealed that patient age at the time of diagnosis was a significant predictor of survival. Another significant predictor of poorer survival was higher tumor grade. Conclusions: Our observations indicate that age and pathological grade can negatively affect survival of individuals with prostate cancer in Iran.

Development of High-yielding Mutants of Streptomyces avermitilis for Avermectin B_{1a} Production through Protoplast Fusion. (원형질체 융합에 의한 Avermectina B_{1a} 고생산성 Streptomyces avermitilis 균주 개발)

  • 김경희;송성기;정연호;정용섭;전계택
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.101-109
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    • 2004
  • In order to enhance the productivity of AVM $B_{la}$ produced by Streptomyces avermitilis as a secondary metabolite, we established a basic protocol necessary for protoplast fusion with high-producing strains as a fusion partner, and then obtained various kinds offusants by adopting a massive strain-development procedure (a miniaturized strain screening system). An alternative fusion method using UV and/or NTG mutation of protoplasts was developed to screen genetic recombinants without specific selectable markers. In this method, the mutants obtained by protoplast fusion after UV and/or NTG treatment (95% death rate) of the respective fusion partner (protoplasts of the respective mutants resistant against L-isoleucine antimetabolites such as O-methylthreonine and/or azaleucine) were regarded as DNA-recombined protoplast fusants. Notably it was demonstrated that most of the protoplast recombinants obtained by the UV mutation method were able to biosynthesize higher amount of AVM $B_{la}$ , reaching almost three times higher level (almost equal to the industrial productivity), compared to the average AVM Bla amount of the parallel mother strains.

Fluorometric Detection of Low-Abundance EGFR Exon 19 Deletion Mutation Using Tandem Gene Amplification

  • Kim, Dong-Min;Zhang, Shichen;Kim, Minhee;Kim, Dong-Eun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.662-667
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    • 2020
  • Epidermal growth factor receptor (EGFR) mutations are not only genetic markers for diagnosis but also biomarkers of clinical-response against tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). Among the EGFR mutations, the in-frame deletion mutation in EGFR exon 19 kinase domain (EGFR exon 19-del) is the most frequent mutation, accounting for about 45% of EGFR mutations in NSCLCs. Development of sensitive method for detecting the EGFR mutation is highly required to make a better screening for drug-response in the treatment of NSCLC patients. Here, we developed a fluorometric tandem gene amplification assay for sensitive detection of low-abundance EGFR exon 19-del mutant genomic DNA. The method consists of pre-amplification with PCR, thermal cycling of ligation by Taq ligase, and subsequent rolling circle amplification (RCA). PCR-amplified DNA from genomic DNA samples was used as splint DNA to conjugate both ends of linear padlock DNA, generating circular padlock DNA template for RCA. Long stretches of ssDNA harboring multiple copies of G-quadruplex structure was generated in RCA and detected by thioflavin T (ThT) fluorescence, which is specifically intercalated into the G-quadruplex, emitting strong fluorescence. Sensitivity of tandem gene amplification assay for detection of the EGFR exon 19-del from gDNA was as low as 3.6 pg, and mutant gDNA present in the pooled normal plasma was readily detected as low as 1% fraction. Hence, fluorometric detection of low-abundance EGFR exon 19 deletion mutation using tandem gene amplification may be applicable to clinical diagnosis of NSCLC patients with appropriate TKI treatment.

Various Renal Manifestations in Children with Tuberous Sclerosis Complex (소아 결절성 경화증에서 나타나는 다양한 신증상들)

  • Lim, Ja Hyun;Park, Moon Sung;Pai, Ki Soo;Kim, Sung Hwan;Shin, Jae Il;Park, Se Jin
    • Childhood Kidney Diseases
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    • v.18 no.2
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    • pp.132-136
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    • 2014
  • Tuberous sclerosis complex (TSC) is a genetic disorder that affects multiple organ systems and causes tumors. It is important that physicians are aware of the manifestations of TSC, and that they follow the recommendations for screening and evaluation. Several types of renal abnormalities may develop in individuals with TSC. Individuals with TSC may require ongoing treatment that can be adapted for each arising manifestation of renal disease. Herein, we report 4 patients with TSC who presented with a range of different renal manifestations, including angiomyolipoma, renal cell carcinoma, renal infarction, renal cyst, and nephrolithiasis.

Molecular Cloning of Differentially Expressed Genes in First Trap Leaf of Dionaea muscipula by Fluorescent Differential Display (형광 Differential Display법에 의한 파리지옥풀 포충잎트랩 특이발현 유전자 탐색)

  • Kang, Kwon-Kyoo;Lee, Keun-Hyang;Park, Jin-Heui;Hong, Kyong-Ei
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.307-313
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    • 2003
  • Fluorescent differential display (FDD) is a method for identifying differentially expressed genes in eukaryotic cells. The mRNA FDD technology works by systematic amplification of the 3' terminal regions of mRNAs. This method involve the reverse transcription using anchored primers designed to bind 5'boundary of the poly A tails, followed by polymerase chain reaction (PCR) amplification with additional upstream primers of arbitrary sequences. The amplified cDNA subpopulations are separated by denaturing polyacrylamide electrophoresis. To identify the genes involved in the development of first trap leaf, we applied a FDD method using mRNAs from leaf base, first trap leaf and flower tissue, respectively. We screened several genes that expressed specifically in first trap leaf. Nucleotide sequence analysis of these genes revealed that these were protease inhibitor (PI), myo-inositol-1-phosphate synthase and lipocalin-type prostaglandin D synthase. Northern blot analysis showed that these genes were expressed specifically in first trap leaf (in vivo and in vitro). FDD could prove to be useful for simultaneous scanning of transcripts from multiple cDNA samples and faster selection of differentially expressed transcripts of interest.

Selection of Starter Cultures and Optimum Conditions for Lactic Acid Fermentation of Onion

  • Choi, You-Jung;Cheigh, Chan-Ick;Kim, Su-Woo;Jang, Jae-Kweon;Choi, Young-Jin;Park, Young-Seo;Park, Hoon;Shim, Kun-Sub;Chung, Myong-Soo
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1100-1108
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    • 2009
  • Lactic acid bacteria (LAB) isolated from various fruits and vegetables were screened in order to determine appropriate fermentation starters for manufacturing functional fermented onion juice. From the initial screening test comprising more than 700 isolated LAB, 16 isolates were selected based on their acid production rate. Among the selected isolates, the fermentation broth of KC-007 exhibited the highest electron donating and nitrite scavenging activities, with values at pH 1.2 of 95.6 and 68.7%, respectively. From the overall results obtained in this study, we finally selected the bacterium KC-007 as a fermentation starter. This bacterium was identified and named as Pediococcus pentosaceus based on its morphological and physiological characteristics, carbon-utilization pattern (as assessed using an API 50CHL kit), and molecular genetic characteristics (as assessed using the nucleotide sequence of the 16S rRNA gene). The optimal temperature, pH, and starter inoculation concentration (v/v) required for growth of the isolated strain were $40^{\circ}C$, pH 4.0-6.0, and 2%(v/v), respectively.

Screening and Characterization of Drosophila Development Mutants Using Single P[en-lacZ] Element Mutagenesis (Drosophila single P[en-lacZ] element mutagenesis를 이용한 발생 관련 돌연변이체 작성)

  • Ha, Hye-Yeong;Lee, Heui-Jung;Park, Soon-Hee;Yoo, Mi-Ae;Lee, Won-Ho
    • Journal of Life Science
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    • v.7 no.1
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    • pp.49-58
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    • 1997
  • Single P[en-lacZ] element including 5.7 kb of engrailed upstream sequences and the E. coli lacZ fusion gene, localized on 48A in rxyho25 strain was transposed to different sites in the Drosophila genome by the jumpstart technique. From 3315 individual genetic crosses, 113 new insertion lines carrying P[en-lacZ] inserted at different sites were obtained. $\beta$-Galactosidase expression in larval tissues of 113 insertion lines were detected by X-gal staining. & among 113 lines have been indentified to be for recessive lethal mutations. Among 7 lines, the #1119 line being lethal during embryogenesis was examined about the ${\beta}$$-Galactosidase expression, nuclear behavior and cellularization pattern during embryogenesis. The P[en-lacZ] insertion lines obtained in this study could be utilized for studying structure and function of the Drosophila development-related genes.

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Evaluation of Native Soybean Collection for Resistance to Purple Blotch (수집재래종대두의 자주빛무늬병(Cercospora kikuchii)에 대한 저항성검정)

  • Oh Jeung Haing;Kwon Shin Han
    • Korean journal of applied entomology
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    • v.20 no.3 s.48
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    • pp.131-134
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    • 1981
  • Native soybean collections were evaluated to search a resistant gene source to purple blotch caused by Cercospora kikuchii. Among 467 native lines, about $28.9\%$ of the lines was less than $0.1\%$ and $13.4\%$ was over $2\%$ in natural infection of purple blotch. Natural infection seemed to be significantly associated with weather conditions at the early podding stage. A significant correlation between natural infection and purple discoloration by seed inoculation was observed and this method seemed to be effective as a preliminary screening technique for resistance to purple blotch. Most of the late maturing native soybeans showed susceptible reaction by the seed inoculation contrary to low infection under natural conditions, indicating that the low natural infection might be due to disease escaping by the late maturing instead of their genetic resistance.

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