• Title/Summary/Keyword: molecular biology method

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Genetic Status of ESR Locus and Other Unidentified Genes As sociated with Litter Size in Chinese Indigenous Tongcheng Pig Breed after a Long Time Selection

  • Zhu, M.J.;Yu, M.;Liu, B.;Zhu, Z.Z.;Xiong, T.A.;Fan, B.;Xu, S.P.;Du, Y.Q.;Peng, Z.Z.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.598-602
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    • 2004
  • The Tongcheng pig breed is a famous Chinese indigenous breed. The Ministry of Agriculture of China has filed it as 1 of 19 national key conservation breeds selected from more than 100 Chinese indigenous pig breeds in 2000. In order to improve the reproductive performance, it has been intensively selected to increase the litter size for about 10 years. The population randomly sampled from conservation nucleus of eight families in the Tongcheng pigs was genotyped for identification of their estrogen receptor locus polymorphisms with the PCR-RFLPs method. Only AB heterozygotes and BB homozygotes were detected, and $X^2$ test demonstrated that the locus was in disequilibrium at a significant level (p<0.05). In the present paper, the litter sizes in different parities were regarded as different traits. Holistic status of other unspecific and unidentified genes was estimated by using the statistical methods. Coefficients of kurtosis and skewness showed that the litter size still presented segregating characteristic in the 2nd, 5th, 7th, 8th and 9th parities. Analysis of homogeneity of variance between families confirmed the results for the 5th, 7th and 8th parities. The heritability of litter size for the 1st to 10th parities was estimated with paternal half-sib model and individual estimated breeding values (EBVs) were evaluated by a single trait animal model as well. We found that the averages of EBVs for litter size in each parity did not differ significantly between genotypes, despite the significant difference for original phenotype records in the 3rd, 4th and 5th parities (p<0.05 or p<0.01). The results may be explained by the deduction that the polymorphisms of ESR locus are no longer the important genetic base of litter size variation when the frequency of allele B accumulated in the experience of selection procedure, and further conferring that there exist special genes associated with litter size in the recent Tongcheng pigs population can be made.

Antitumor Responses of Adoptively-Transferred Tumor-Specific T-Cell Cultures in a Murine Lymphoma Model

  • Kim, Hee-Sue;Lee, Hee-Gu;Lim, Jong-Seok;Lee, Ki-Young;Kim, Jae-Wha;Chung, Kyeong-Soo;Choe, Yong-Kyung;Choe, In-Seong;Chung, Tai-Wha;Kim, Kil-Hyoun
    • BMB Reports
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    • v.28 no.6
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    • pp.556-561
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    • 1995
  • The purpose of this study was to establish an in vitro culture method of tumor-specific T cells, and determine the efficacy of the cultured tumor-specific cytotoxic T-lymphocytes (CTL) as an agent of anti-tumor immunotherapy against a murine lymphoma, TIMI.4. Tumor-specific T-lymphocytes derived from C57BL/6 mice (thy-1.2) immune to TIMI.4 were activated by in vitro stimulation with the irradiated TIMI.4 cells, and expanded by restimulation with TIMI.4 in the presence of the concanavalin A-stimulated rat spleen culture supernatant, and splenic antigen-presenting cells. In vitro restimulation enhanced markedly the proportion of $CD8^+$, a predominant surface marker of CTL and the cytotoxic activity in the cultured immune T cell population. The resulting TIMI.4-specific T cells were adoptively transferred into nude mice. The tumor cells residing in the host after 7 days of adoptive transfer to B6.PL (thy-1.1) mice were quantified by use of an antibody directed to the thy-1.2 allele. The TIMI.4 cells in the recipient nude mice were decreased in a dose-dependent manner. Anti-tumor activity of the TIMI.4-specific T cells was also demonstrated by a survival test, where the tumor-bearing nu/nu mice which received the activated T-cells survived about 30% longer than the control mice which received the tumor cells alone. These suggest that adoptive transfer of TIMI.4-specific T cells could be a candidate for effective therapy of the murine lymphoma.

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Determination of the Number of Components in Spectroscopy from the Multilinear Model Fitting

  • Kim, Choong-Rak;Chung, Byung-Chull;Lee, Choon-Hwan
    • Communications for Statistical Applications and Methods
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    • v.6 no.2
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    • pp.367-374
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    • 1999
  • Biological specimens contain several components and multilinear models are very useful in analyzing these data. After fitting the model the number of components are determined by the change of mean squared error however this method is quite rule of thumb. in this paper we suggest a measure to decide the number of components based on the relative change of to mean squared error. Simulations are done and applications to real data set are given as illustrations.

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Greenhouse Method for Assessing Spot Blotch Resistance in Barley

  • Arabi, Mohammad Imad Eddin;Jawhar, Mohammad
    • The Plant Pathology Journal
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    • v.26 no.4
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    • pp.421-423
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    • 2010
  • New sources of barley (Hordeum vulgare L.) resistant to spot blotch, caused by Cochliobolus sativus, are needed to provide effective resistance because of the rapid change pathotype patterns of C. sativus in fields. The purposes of our study were to develop a method to screen barley for resistance to spot blotch disease and then use this methodology to screen barley genotypes for resistance to the major virulent pathotype Pt4 in barley populations in Syria. A transparent tape method, in which a conidial suspension of C. sativus was dropped onto transparent tape and placed, treated-side down, on the second leaf surface of barley plants. Disease symptoms of fungus were easily detected on the leaves covered by the transparent tape after 48h of inoculation. The transparent tape method was repeatable and the disease scores obtained were correlated (r = 0.91, P = 0.001) with those obtained by the seedling assay. This method may be beneficial in various plant pathology breeding programs.

Nucleotide Analysis of 185 rRNA and Molecular Phylogeny of the Korean Decapods (하국산 십각류의 18S 리보솜 RNA의 염기분석과 분자계통에 관한 연구)

  • Kim, Won
    • The Korean Journal of Zoology
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    • v.35 no.1
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    • pp.80-86
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    • 1992
  • The nucleotide sequences of 185 rRNAs of the five Korean decapods were partially determined by the direct sequencing method using the reverse transcriptase. ne average GC content of five species was 51.1% which is higher than that of yeast(45.0%) and lower than those of frog (53.0%) and rat (55.6%). This result follows the general patterns of the GC content in the nucleotides of the nucleic acid shown among the various phylogenetic groups. The average ratio of transrional/transversional nucleotide substitution of pairwise comparison among six species (including Anemia salina) was 1.200 $\pm$ 0.310 when whole region alas examined. However, the ratio showed some differences when the conservative regions and variable regions frere separatelv examined. The molecular phylogenies of the five species were constructed by using two different tree making methods. In general the results support the previously reported molecular phylogeny of the decapod crustaceans. However, our results indicate thats in the analysis of the sequence dat3, the UPGMA clustering method of the distance matrix method should be carefully employed after considering the rate of nucneotide substitution in the different regions of the molecule.

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In vivo Monitoring of the Incorporation of Chemicals into Cucumber end Rice Leaves by Chlorophyll Fluorescence Imaging

  • Kim, Jin-Hong;Jung, Ji-Eun;Lee, Choon-Hwan
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.171-178
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    • 2002
  • Chlorophyll (Chl) fluorescence imaging was used to investigate the effectiveness of in vivo incorporation methods for two chemicals, 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU) and methyl viologen (MV) in rice, a monocot, and cucumber, a dicot, leaves. four different methods (vacuum infiltration, floating, transpiration-aided incorporation through petiole and spraying) were compared, and $F_i$ and $F_v$/$F_m$ were chosen for the imaging of the DCMU- and MV-treated leaves, respectively. The effects of the chemicals in plants were generally heterogeneous over the whole leaf area. Moreover, the effectiveness of the treatment of a chemical in plant leaves was dependent on chemical species, plant species, concentration of the chemical, the treatment method, the duration of the treatment, the existence of light and detergent, etc. In conclusion, we suggest that to achieve the proposed effects of chemicals in plants for an actual experiment, these factors must be considered before the chemical treatment, and the best method for the treatment of the chemicals tested was floating and vacuum infiltration in the dicot and the monocot leaves, respectively, as drawn from Chl fluorescence imaging analysis.

Enhanced Drug Carriage Efficiency of Curcumin-Loaded PLGA Nanoparticles in Combating Diabetic Nephropathy via Mitigation of Renal Apoptosis

  • Asmita Samadder;Banani Bhattacharjee;Sudatta Dey;Arnob Chakrovorty;Rishita Dey;Priyanka Sow;Debojyoti Tarafdar;Maharaj Biswas;Sisir Nandi
    • Journal of Pharmacopuncture
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    • v.27 no.1
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    • pp.1-13
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    • 2024
  • Background: Diabetic nephropathy (DN) is one of the major complications of chronic hyperglycaemia affecting normal kidney functioning. The ayurvedic medicine curcumin (CUR) is pharmaceutically accepted for its vast biological effects. Objectives: The Curcuma-derived diferuloylmethane compound CUR, loaded on Poly (lactide-co-glycolic) acid (PLGA) nanoparticles was utilized to combat DN-induced renal apoptosis by selectively targeting and modulating Bcl2. Methods: Upon in silico molecular docking and screening study CUR was selected as the core phytocompound for nanoparticle formulation. PLGA-nano-encapsulated-curcumin (NCUR) were synthesized following standard solvent displacement method. The NCUR were characterized for shape, size and other physico-chemical properties by Atomic Force Microscopy (AFM), Dynamic Light Scattering (DLS) and Fourier-Transform Infrared (FTIR) Spectroscopy studies. For in vivo validation of nephro-protective effects, Mus musculus were pre-treated with CUR at a dose of 50 mg/kg b.w. and NCUR at a dose of 25 mg/kg b.w. (dose 1), 12.5 mg/kg b.w (dose 2) followed by alloxan administration (100 mg/kg b.w) and serum glucose levels, histopathology and immunofluorescence study were conducted. Results: The in silico study revealed a strong affinity of CUR towards Bcl2 (dock score -10.94 Kcal/mol). The synthesized NCUR were of even shape, devoid of cracks and holes with mean size of ~80 nm having -7.53 mV zeta potential. Dose 1 efficiently improved serum glucose levels, tissue-specific expression of Bcl2 and reduced glomerular space and glomerular sclerosis in comparison to hyperglycaemic group. Conclusion: This study essentially validates the potential of NCUR to inhibit DN by reducing blood glucose level and mitigating glomerular apoptosis by selectively promoting Bcl2 protein expression in kidney tissue.

Molecular identification of selected parrot eggs using a non-destructive sampling method

  • Jung-Il Kim;Jong-Won Baek;Chang-Bae Kim
    • Korean Journal of Environmental Biology
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    • v.41 no.2
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    • pp.145-166
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    • 2023
  • Parrots have been threatened by global trade to meet their high demand as pets. Controlling parrot trade is essential because parrots play a vital role in the ecosystem. Accurate species identification is crucial for controlling parrot trade. Parrots have been traded as eggs due to their advantages of lower mortality rates and more accessible transport than live parrots. A molecular method is required to identify parrot eggs because it is difficult to perform identification using morphological features. In this study, DNAs were obtained from 43 unidentified parrot eggs using a non-destructive sampling method. Partial cytochrome b (CYTB) gene was then successfully amplified using polymerase chain reaction (PCR) and sequenced. Sequences newly obtained in the present study were compared to those available in the GenBank by database searching. In addition, phylogenetic analysis was conducted to identify species using available sequences in GenBank along with sequences reported in previous studies. Finally, the 43 parrot eggs were successfully identified as seven species belonging to two families and seven genera. This non-destructive sampling method for obtaining DNA and molecular identification might help control the trade of parrot eggs and prevent their illegal trade.

A Simple Method for Assessing Severity of Common Root Rot on Barley

  • Arabi, Mohammad Imad Eddin;Jawhar, Mohammad
    • The Plant Pathology Journal
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    • v.29 no.4
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    • pp.451-453
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    • 2013
  • Common root rot caused by Cochliobolus sativus is a serious disease of barley. A simple and reliable method for assessing this disease would enhance our capacity in identifying resistance sources and developing resistant barley cultivars. In searching for such a method, a conidial suspension of C. sativus was dropped onto sterilized elongated subcrown internodes and incubated in sandwich filter paper using polyethylene transparent envelopes. Initial disease symptoms were easily detected after 48h of inoculation. Highly significant correlation coefficients were found in each experiment (A, B and C) between sandwich filter paper and seedling assays, indicating that this testing procedure was reliable. The method presented facilitates a rapid pre-selection under uniform conditions which is of importance from a breeder's point of view.

Multiple Age-Associated Mitochondrial DNA Deletions in Mouse Brain

  • Kim, Jin-Sun;Kim, Min-Jung;Kwon, In-Sook;Song, Eun-Sook
    • BMB Reports
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    • v.30 no.1
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    • pp.33-36
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    • 1997
  • Age-dependent deletion of mitochondrial DNA (mtDNA) was detected in mouse brain using PCR method. The size of the deleted fragment was 0.5 kb, 0.9 kb. 1.7 kb and 4.3 kb in the region between cytochrome b gene and ATPase 6 gene. The deleted fragment was increased gradually from 3-month to 22month Direct repeat sequence flanking the deletion in 0.5 kb PCR product was TAAT.

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