• Title/Summary/Keyword: Genetic evaluation

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Comparison of In Vitro Cell Transformation Assay Using Murine Fibroblasts and Human Keratinocytes

  • Ahn, Jun-Ho;Park, Sue-Nie;Yum, Yung-Na;Kim, Ji-Young;Lee, Michael
    • Toxicological Research
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    • v.24 no.1
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    • pp.37-44
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    • 2008
  • The in vitro cell transformation assays (CTA) were performed using BALB/3T3 murine fibroblasts and HaCaT human keratinocytes in order to evaluate concordance between both in vitro CTAs and carcinogenicity with compounds differing in their genotoxic and carcinogenic potential. Six test articles were evaluated, two each from three classes of compounds: genotoxic carcinogens (2-amino-5-nitrophenol and 4-nitroquinoline-N-oxide), genotoxic noncarcinogens (8-hydroxyquinoline and benzyl alcohol), and nongenotoxic carcinogens (methyl carbamate and N-nitrosodiphenylamine). Any foci of size $\geq$2 mm regardless of invasiveness and piling was scored as positive in CTA with BALB/3T3. As expected, four carcinogens regardless of their genotoxicity had positive outcomes in two-stage CTA using BALB/3T3 cells. However, of the two genotoxic noncarcinogens, benzyl alcohol was positive CTA finding. We concluded that, of the 6 chemicals tested, the sensitivity for BALB/3T3 system was reasonably high, being 100%. The respective specificity for BALB/3T3 assay was 50%. We also investigated the correlation between results of BALB/3T3 assay and results from HaCaT assay in order to develop a reliable human cell transformation assay. However, evaluation of staining at later time points beyond the confluency stage did not yield further assessable data because most of HaCaT cells were detached after $2{\sim}3$ days of confluency. Thus, after test article treatment, HaCaT cells were split before massive cell death began. In this modified protocol for this HaCaT system, growing attached colonies were counted instead of transformed foci 3 weeks since last subculture. Compared to BALB/3T3 assay, HaCaT assay showed moderate low sensitivity and high specificity. Despite these differences in specificity and sensitivity, both cell systems did exhibit same good concordance between in vitro CTA and rodent carcinogenicity findings (overall 83% concordant results). At present the major weakness of these in vitro CTA is lack of validation for regulatory acceptance and use. Thus, more controlled studies will be needed in order to be better able to assess and quantitatively estimate in vitro CTA data.

Genetic Diversity and Pathotypes of Xanthomonas orzyae pv. oryzae Isolated in Korea (국내 수집 벼흰잎마름병균의 유전적 다양성 및 병원형)

  • Oh, Chang-Sik;Roh, Eun-Jung;Lee, Seung-Don;La, Dong-Soo;Heu, Sung-Gi
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.224-231
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    • 2010
  • Xanthomonas oryzae pv. oryzae, causal agent of bacterial leaf blight (BLB) of rice, had been collected and identified using Biolog and fatty acid analysis. Epidemics of BLB had been occurred all the times at several rice cultivating areas in Korea in 1999-2004. Most X. oryzae pv. oryzae isolated in 1999 and 2002 belonged to Korean race K1, but more than 50% of the pathogen isolated in 2003 belonged to Korean race K3. Especially, most pathogens isolated in Jeonnam and Joenbuk provinces belonged to Korean race K3. Inoculation test of near isogenic lines (NIL) of rice carrying single resistance genes against BLB showed that many isolates belonging to Korean race 1 reacted differently to diverse resistant monogenic lines of rice. Southern blot analysis also showed that the bacterial pathogens belonged to the same race had different numbers of avirulence genes. This results suggested that each Korean race type may respond to many resistance genes of rice. All the K3 races isolated in Jeonnam and Joenbuk provinces were able to cause disease on Xa3 monogenic lines of rice. Since most rice cultivars cultivated in Jeonnam and Jeonbuk were carrying Xa3 resistance genes, the bacterial pathogens isolated in Jeonnam and Jeonbuk were likely to develop to adapt to Xa3 resistance gene. Together with avirulence gene patterns of the bacterial isolates and the results of disease reaction of monogenic lines of rice to them, Korean X. oryzae pv. oryzae was classified into 19 pathotypes. This newly classified pathotypes should help the breeding of new resistance rice cultivars in Korea.

Evaluation of resistance to Pierce's disease among grapevine cultivars by using the culture filtrates produced from Xylella fastidiosa (Xylella fastidiosa의 배양여액을 이용한 포도나무 피어스병 품종 저항성 검정)

  • Park, Myung Soo;Lu, Jiang;Yun, Hae Keun
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.394-400
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    • 2017
  • This study investigated whether culture filtrates produced by Xylella fastidiosa can be used to determine the varietal susceptibility to Pierce's disease in grapevines (Vitis spp.) as a substitute for pathogen inoculation or field screening. A bioassay of grape leaves with culture filtrates from the pathogen showed that their phytotoxicities were active and host-selective. Ethyl acetate extracts from them also showed toxicities and host selectivity in both bunches of grapes and muscadine grapes. The sensitive range of plants to the culture filtrates and their ethyl acetate extracts was consistent with the host range of the Pierce's disease pathogen. Susceptible cultivars are sensitive to even highly diluted culture filtrates, while resistant cultivars were not affected even at their original culture filtrates. Susceptible cultivars were more sensitive to the undiluted culture filtrate than were highly diluted culture filtrates, and the younger leaves were the most sensitive to the culture filtrates in grapes. Although some European grape cultivars showed moderately susceptibility in this study, the determination of varietal resistance to Pierce's disease by the treatment of culture filtrates of pathogens could provide valuable information for the preliminary selection of genetic resources and seedlings from hybridization in a disease resistant grape breeding program.

Effects of Body Condition Score and Estimation of Growth Curves for Chest Girth and Ultra Sonic Longissimus Muscle Area, Backfat Thickness and Marbling Scores in Hanwoo(Korean cattle) Cows (한우 암소의 흉위, 초음파 측정 배장근단면적, 등지방두께, 근내지방도에 대한 발육곡선 추정 및 신체충실지수 효과)

  • Lee, Deuk-Hwan;Lee, Gil-Hwan;Cho, Chung-Il;Kim, Nae-Soo
    • Journal of Animal Science and Technology
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    • v.50 no.5
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    • pp.581-590
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    • 2008
  • Growth curves for ultrasonic carcass traits such as longissimus muscle area, backfat thickness and marbling score as well as chest girth which was simultaneously measured when carcass traits were investigated using ultrasound measuring technique were estimated to identify growth patterns and to adjust maturing effects in order to evaluating genetic merits on cows in farming basis. 27,410 records from 22,451 cows on which of 15~90 month of age were investigated from the national wide of Korea using by ultrasonic scanning techniques by the skilled persons from 2002 to 2007. Van Bertalanffy growth function was applied for estimating growth curves on these traits. Carcass traits and chest girth would be linearly increased by body condition score. It might be used for multiplicative correction factors for pre- adjustment on the body condition scores. Growth pattern on chest girth would be quickly reached to mature size and stable on after reached to asymptotic mature size. Longissimus muscle area would also be reached to mature size but little smoother than chest girth. Otherwise, growth curve on backfat thickness would be steadily increasing up to 7 years of age. It also showed large individual difference by way of mean square error. Marbling score would be steadily increased but sharper than those on backfat thickness. It would be reached to mature size up at 5 years of age. Those growth curves would be used for correcting function on age at investigating on genetic evaluation system.

Utility of a multiplex reverse transcriptase-polymerase chain reaction assay (HemaVision) in the evaluation of genetic abnormalities in Korean children with acute leukemia: a single institution study

  • Kim, Hye-Jin;Oh, Hyun Jin;Lee, Jae Wook;Jang, Pil-Sang;Chung, Nack-Gyun;Kim, Myungshin;Lim, Jihyang;Cho, Bin;Kim, Hack-Ki
    • Clinical and Experimental Pediatrics
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    • v.56 no.6
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    • pp.247-253
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    • 2013
  • Purpose: In children with acute leukemia, bone marrow genetic abnormalities (GA) have prognostic significance, and may be the basis for minimal residual disease monitoring. Since April 2007, we have used a multiplex reverse transcriptase-polymerase chain reaction tool (HemaVision) to detect of GA. Methods: In this study, we reviewed the results of HemaVision screening in 270 children with acute leukemia, newly diagnosed at The Catholic University of Korea from April 2007 to December 2011, and compared the results with those of fluorescence in situ hybridization (FISH), and G-band karyotyping. Results: Among the 270 children (153 males, 117 females), 187 acute lymphoblastic leukemia and 74 acute myeloid leukemia patients were identified. Overall, GA was detected in 230 patients (85.2%). HemaVision, FISH, and G-band karyotyping identified GA in 125 (46.3%), 126 (46.7%), and 215 patients (79.6%), respectively. TEL-AML1 (20.9%, 39/187) and AML1-ETO (27%, 20/74) were the most common GA in ALL and AML, respectively. Overall sensitivity of HemaVision was 98.4%, with false-negative results in 2 instances: 1 each for TEL-AML1 and MLL-AF4. An aggregate of diseases-specific FISH showed 100% sensitivity in detection of GA covered by HemaVision for actual probes utilized. G-band karyotype revealed GA other than those covered by HemaVison screening in 133 patients (49.3%). Except for hyperdiplody and hypodiploidy, recurrent GA as defined by the World Health Organizationthat were not screened by HemaVision, were absent in the karyotype. Conclusion: HemaVision, supported by an aggregate of FISH tests for important translocations, may allow for accurate diagnosis of GA in Korean children with acute leukemia.

Clinical Features and Genetic Analysis of Homocystinuria Patients in Korea (전형적 호모시스틴뇨증 환자들의 임상적 특성과 유전자분석)

  • Lee, Yena;Lee, Jeongho;Lee, Dong Hwan
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.15 no.2
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    • pp.78-86
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    • 2015
  • Purpose: Homocystinuria (OMIM#236200) is a metabolic disease caused by mutation in the CBS gene. This study was conducted to identify the clinical features and prognosis of homocystinuria as well as to find out the CBS gene mutations of the six homocystinuria patients who were receiving treatment in the Pediatric Department at Soonchunhyang University Hospital. Methods: From January 1992 to March 2015, clinical, biochemical, and genetic analyses were performed retrospectively on the six patients diagnosed with classic homocystinuria at Soonchunhyang University Hospital. Results: A total of six patients were included in this study, including three who were diagnosed with homocystinuria at the mean age of $50{\pm}22.5$ days based on their abnormal newborn screening test results. The other three were diagnosed at the mean age of 7, when they visited the hospital for evaluation of developmental delay and lens dislocation. The group diagnosed at early infancy had normal cognitive function, but the other group had varying degrees of mental retardation. Major complications were found only in the group diagnosed after infancy. CBS gene mutation was found in all the patients, and they were all non-responsive to vitamin B6 treatment. At present, all patients' diets are controlled following a methionine-free formula and they are on medication with folic acid, betaine, pyridoxine, and methylcobalamin. Conclusion: Six homocystinuria patients were monitored for up to 23 years. The group diagnosed at early infancy exhibited no major complications. Therefore, early diagnosis is crucial in the prognosis, and homocystinuria must be included in the newborn screening program.

Evaluation of the Genetic Diversities and the Nutritional Values of the Tra (Pangasius hypophthalmus) and the Basa (Pangasius bocourti) Catfish Cultivated in the Mekong River Delta of Vietnam

  • Men, L.T.;Thanh, V.C.;Hirata, Y.;Yamasaki, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.671-676
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    • 2005
  • A total of 50 individual catfish, the Tra (Pangasius hypophthalmus) cultivated in either floating cages (Tra-c) or in ponds (Tra-p) and the Basa (Pangasius bocourti) raised in three floating cages, were collected in two of the Mekong Delta provinces. The caudal fin of each individual fish was used for protein electrophoresis employing the SDS-PAGE method. The one fillet sides were used as a representative sample to determine the dry matter (DM), crude protein (CP), ether extract (EE) and amino acids (AAs). The catfish oil was extracted from the belly fats, and the fatty acid (FA) composition was analyzed. There were 21 bands of the Tra and the Basa. Protein bands of the two varieties were 28.6-33.3% polymorphic, while polymorphic individuals of the Tra ranged from 80.0 to 100.0%, and the Basa was 90.0% polymorphic. The phenotypic diversity (Ho) of the Tra ranged from 1.71 to 1.80, while the Basa ranged as high as 2.14%. Diversity values (H$_{EP}$) for genetic diversity markers were equal in the Tra and the Basa. The sum of the effective number of alleles (SENA) of both varieties ranged from 3.40 to 3.83 for the Basa and the Tra, respectively. The lower values of Ho and SENA, as compared with those of the fresh water prawn (Macrobrachium equidens) in the area, would suggest that the species with the low values will become extinct due to inbreeding; the gene pools of each observed population were below a suitable threshold. Many of the differences in the nutritional values of the Tra-c, the Tra-p and the Basa were measured; their nutrient values were comparable to fishmeal or fish oil. Most of the DM, CP, and EE were higher in the Tra, especially in the Tra-c. The essential AA content, especially that of lysine, was highest in the Tra-c, next highest in the Tra-p, and lowest in the Basa. Therefore, the amino acid patterns were closer to the ideal patterns in the same sequences. In contrast, the essential FAs were concentrated in the Basa fish oil. It was found that suitable selection of parents for seed production is required to avoid inbreeding. Catfish may be valuable sources of nutrition for both humans and animals, and the differences in their nutritional values by variety and/or management must be taken into account.

Flock composition, breeding strategies and farmers' traits of interest evaluation of Wollo highland sheep and their F1 crosses

  • Amare, Tadesse;Goshu, Gebeyehu;Tamir, Berhan
    • Journal of Animal Science and Technology
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    • v.60 no.5
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    • pp.14.1-14.12
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    • 2018
  • Background: Sheep production is a major component of the livestock sector in Ethiopia. The country owing to the large population of 30.70 million estimated numbers of sheep in the country and out of which about 72.14% are females, and 27.86% are males with diverse genetic resources. The real value of indigenous breeds was often under-estimated mostly due to their poor appearance and relatively low productivity. Developing countries in most cases opt for exotic breeds to increase animal productivity through crossbreeding or breed substitution without properly investigating the production potential of the indigenous breeds. The main objective of the research was to identify sheep flock composition and structure, farmers' traits of interest and breeding objective of Wollo highland sheep, and their $F_1$ crossbreed progenies. Results: Smallholder farmers' flock synthesized from breeding ewes, breeding rams, pre-weaned ewe lambs, pre-weaned ram lambs, unproductive ewes, castrated and fattened rams, with the percentage coverage of 29.2, 13.3, 15.5, 16.5, 12.4, and 12.5%, respectively. The maximum number of flock size was 289.0 sheep per flock and higher in the third stratum. The off-take rate percentage of the three strata presented as 21.9% in 1st stratum, 12% in the 2nd stratum, and 16.4% in the 3rd stratum and higher off-take rate recorded in the first stratum. Sheep producer's traits of interest ranked by growth rate (first), body size (second) and marketing value was third rank. Communal breeding (random mating), village based controlled breeding, mixed type and private ram controlled breeding practice were comprised of 39.7, 61.7, 52 and 71.3%, respectively. The percentages of ewes per flock composition were presented as 36.5, 27.1 and 25.5%, respectively in the 3rd stratum, 2nd stratum and 3rd stratum in the order of their importance's. Conclusion: Genetic improvement practices at smallholder sheep producers situation was showing promising outcome with indigenous Washera $F_1$ crossbred lambs and which designated for weaning rate, body size, marketing age, age at first lambing, good temperament and large litter size in the order of their rank. The contemporary breeding practice tendency indicated that, reduced flock size to improve flock productivity via crossbreeding practices.

Evaluation of General Toxicity and Genotoxicity of the Silkworm Extract Powder

  • Heo, Hyun-Suk;Choi, Jae-Hun;Oh, Jung-Ja;Lee, Woo-Joo;Kim, Seong-Sook;Lee, Do-Hoon;Lee, Hyun-Kul;Song, Si-Whan;Kim, Kap-Ho;Choi, Yang-Kyu;Ryu, Kang-Sun;Kang, Boo-Hyon
    • Toxicological Research
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    • v.29 no.4
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    • pp.263-278
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    • 2013
  • The silkworm extract powder contain 1-deoxynojirimycin (DNJ), a potent ${\alpha}$-glycosidase inhibitor, has therapeutic potency against diabetes mellitus. Therefore, natural products containing DNJ from mulberry leaves and silkworm are consumed as health functional food. The present study was performed to evaluate the safety of the silkworm extract powder, a health food which containing the DNJ. The repeated toxicity studies and gentic toxicity studies of the silkworm extract powder were performed to obtain the data for new functional food approval in MFDS. The safety was evaluated by a single-dose oral toxicity study and a 90 day repeated-dose oral toxicity study in Sprague-Dawley rats. The silkworm extract powder was also evaluated for its mutagenic potential in a battery of genetic toxicity test: in vitro bacterial reverse mutation assay, in vitro chromosomal aberration test, and in vivo mouse bone marrow micronucleus assay. The results of the genetic toxicology assays were negative in all of the assays. The approximate lethal dose in single oral dose toxicity study was considered to be higher than 5000 mg/kg in rats. In the 90 day study, the dose levels were wet at 0, 500, 1000, 2000 mg/kg/day, and 10 animals/sex/dose were treated with oral gavage. The parameters that were monitored were clinical signs, body weights, food and water consumptions, ophthalmic examination, urinalysis, hematology, serum biochemistry, necropsy findings, organ weights, and histopathological examination. No adverse effects were observed after the 90 day administration of the silkworm extract powder. The No-Observed-Adverse-Effect-Level (NOAEL) of silkworm extract powder in the 90 day study was 2000 mg/kg/day in both sexes, and no target organ was identified.

A Missense Mutation in Exon 5 of the Bovine Growth Hormone Gene (소 성장호르몬 유전자의 Exon 5번에서의 새로운 다형성 연구)

  • Yoon, D. H.;Kim, T. H.;Lee, K. H.;Park, E. W.;Lee, H. K.;Cheong, I. C.;Hong, K. C.
    • Journal of Animal Science and Technology
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    • v.45 no.1
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    • pp.13-22
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    • 2003
  • Growth Hormone (GH) gene is a member of gene family through the evolutionary process from a small common ancestral gene by a series of gene duplications. The role of the GH in growth and performance controls has been extensively studied in human, mice and livestock. Many researchers have considered GH as a strong candidate gene for evaluation of genetic polymorphisms that could be associated with economic traits in cattle. We report here a novel missense mutation within the exon 5 of the bovine Growth Hormone (bGH) gene. We could amplified 522 bp fragments from eight unrelated Hanwoo cattle by PCR, then, subsequently cloned and sequenced. An Msp I RFLP corresponding to a C to T transition was observed at position 2258 nt. From this result, we could predict a missense mutation (Arg to Trp) at codon 166 in a highly conserved region among many mammals. Codominant Mendelian segregation of the two alleles, Msp I (+) and Msp I (-), was observed in two full-sib F2 families (n = 32, African taurine Bos taurus ${\times}$ African zebu Bos indicus) and eight half-sib Hanwoo families. For the availability of genetic marker, we have performed PCR-RFLP with a large number of individual animals from 15 different cattle breeds (European and Asian taurines, and African indicines). Consideration of breed frequencies of Msp I (-) allele in relation to breed type and their geographic origins, shows higher frequencies in humped breeds or Asian cattle breeds than in humpless or European breeds. This result indicates that the missense mutation can be contributed the functional significance such as the signal transduction through the receptor binding, also may be used as a marker for selection of the economic traits in Hanwoo.