Park C. S.;Li Z. H.;Sung N. D.;Jin D. I.;Cong P. Q.;Kim E. S.;Song E. S.;Yi Y. J.
Reproductive and Developmental Biology
/
v.29
no.4
/
pp.253-257
/
2005
This study was carried out to evaluate the effects of washing medium, breed and washing temperature of fresh and frozen-thawed boar sperm on mitochondrial activity and membrane integrity by flow cytometry. More than $80\%$ of fresh sperm washed with mTLP-PVA medium at $20^{\circ}C$ exhibited an intact membrane and a functional mitochondrion. With frozen-thawed samples, a large number of sperm showed both damaged membrane $(36.4\~46.9\%)$ and nonfunctional mitochondrion $(55.1\~71.1\%)$ in the mTLP-PVA and BTS washing media at $20^{\circ}C$. There were no breed effects of fresh and frozen-thawed sperm on mitochondrial activity and membrane integrity. The percentages of damaged membrane of fresh and frozen sperm, respectively, were higher at $4^{\circ}C$ washing temperature than at $20^{\circ}C$ washing temperature in the mTLP-PVA medium. We found that washing medium and washing temperature of fresh and frozen-thawed boar sperm were important for the analyses of mitochondrial activity and membrane integrity by flow cytometry.
Journal of Fisheries and Marine Sciences Education
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v.29
no.2
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pp.513-525
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2017
In the estimation of the exploitable carrying capacity (ECC) in the Korean water of the East China Sea, two approaches, which are the ecosystem modeling method (EMM) and the holistic production method (HPM), were applied. The EMM is accomplished by Ecopath with Ecosim model using a number of ecological data and fishery catch for each species group, which was categorized by a self-organizing mapping (SOM) based on eight biological characteristics of species. In this method, the converged value during the Ecosim simulation by setting the instantaneous rate of fishing mortality (F) as zero was estimated as the ECC of each group. The HPM is to use surplus production models for estimateing ECC. The ECC estimates were 4.6 and 5.1 million mt (mmt) from EMM and HPM, respectiverly. The estimate from the EMM has a considerable uncertainty due to the lack of confidence in input ecological parameters, especially production/biomass ratio (P/B) and consumption/biomass ratio (Q/B). However, ECC from the HPM was estimated on the basis of relatively fewer assumptions and long time-series fishery data as input, so the estimate from the HPM is regarded as more reasonable estimate of ECC, although the ECC estimate could be considerd as a preliminary one. The quality of input data should be improved for the future study of the ECC to obtain more reliable estimate.
The present study was conducted to investigate the genetic characteristic and to establish the parentage verification system of the Korean native horse(KNH). A total number of 192 horses from six horse breeds including the KNH were genotyped using 17 microsatellite loci. This method consisted of multiplexing PCR procedure. The number of alleles per locus varied from 5 to 10 with a mean value of 7.35 in KNH. The expected heterozygosity and observed heterozygosity were ranged from 0.387 to 0.841(mean 0.702) and from 0.429 to 0.905(mean 0.703), respectively. The total exclusion probability of 17 microsatellite loci was 0.9999. Of the 17 markers, AHT4, AHT5, CA425, HMS2, HMS3, HTG10, LEX3 and VHL20 marker have relatively high PIC value(>0.7). This study found that there were specific alleles, P allele at AHT5, Q allele and R allele at ASB23, H allele at CA425, S allele at HMS3, J allele at HTG10 and J allele at LEX3 marker in KNH when compared with other horse populations. Also, the results showed two distinct clusters: the Korean native horse cluster(Korean native horse, Mongolian horse), and the European cluster(Jeju racing horse, Thoroughbred horse). These results present basic information for detecting the genetic markers of the KNH, and has high potential for parentage verification and individual identification of the KNH.
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.
Soybean transgenic plants with ectopically expressed AtABF3 were produced by Agrobacterium-mediated transformation and investigated the effects of AtABF3 expression on drought and salt tolerance. Stable Agrobacterium-mediated soybean transformation was carried based on the half-seed method (Paz et al. 2006). The integration of the transgene was confirmed from the genomic DNA of transformed soybean plants using PCR and the copy number of transgene was determined by Southern blotting using leaf samples from $T_2$ seedlings. In addition to genomic integration, the expression of the transgenes was analyzed by RT-PCR and most of the transgenic lines expressed the transgenes introduced. The chosen two transgenic lines (line #2 and #9) for further experiment showed the substantial drought stress tolerance by surviving even at the end of the 20-day of drought treatment. And the positive relationship between the levels of AtABF3 gene expression and drought-tolerance was confirmed by qRT-PCR and drought tolerance test. The stronger drought tolerance of transgenic lines seemed to be resulted from physiological changes. Transgenic lines #2 and #9 showed ion leakage at a significantly lower level (P < 0.01) than ${\underline{n}}on-{\underline{t}}ransgenic$ (NT) control. In addition, the chlorophyll contents of the leaves of transgenic lines were significantly higher (P < 0.01). The results indicated that their enhanced drought tolerance was due to the prevention of cell membrane damage and maintenance of chlorophyll content. Water loss by transpiration also slowly proceeded in transgenic plants. In microscopic observation, higher stomata closure was confirmed in transgenic lines. Especially, line #9 had 56% of completely closed stomata whereas only 16% were completely open. In subsequent salt tolerance test, the apparently enhanced salt tolerance of transgenic lines was measured in ion leakage rate and chlorophyll contents. Finally, the agronomic characteristics of ectopically expressed AtABF3 transgenic plants ($T_2$) compared to NT plants under regular watering (every 4 days) or low rate of watering condition (every 10 days) was investigated. When watered regularly, the plant height of drought-tolerant line (#9) was shorter than NT plants. However, under the drought condition, total seed weight of line #9 was significantly higher than in NT plants (P < 0.01). Moreover, the pods of NT plants showed severe withering, and most of the pods failed to set normal seeds. All the evidences in the study clearly suggested that overexpression of the AtABF3 gene conferred drought and salt tolerance in major crop soybean, especially under the growth condition of low watering.
The molecular mechanisms involved in the progression of clear cell renal cell carcinomas (ccRCCs) are still unclear. The aim of this study was to analyse the relationships between expression of RALYL and clinical characteristics. In 41 paired samples of ccRCCs and adjacent normal tissues, we used real-time qPCR to evaluate the expression of RALYL mRNA. RALYL protein levels were determined in 146 samples of ccRCC and 37 adjacent normal tissues by immunohistochemistry. Statistical analysis was used to explore the relationships between expression of RALYL and the clinical characteristics (gender, age, tumor size, T stage, N stage, M stage, survival times and survival outcome) in ccRCC. In addition, these patients were follow-up period 64 months (range: 4~116months) to investigate the influence on prognosis. We found significantly differences between ccRCC tissues and normal tissues (p<0.001, paired-sample t test) in mRNA levels of RALYL. Immunohistochemistry analyses in 146 ccRCC samples and 37 adjacent normal tissues showed significantly lower RALYL protein levels in ccRCC samples (${\chi}^2$-test, p<0.001), inversely correlating with tumour size (p=0.024), T stage (0.005), N stage (p<0.001) as well as M stage (p=0.019), but not age (p=0.357) and gender (p=0.348). Kaplan-Meier survival analysis demonstrated that people with lower level of RALYL expression had a poorer survival rate than those with a higher level of RALYL expression, significantly different by the log-rank test (p=0.011). Cox regression analysis indicated that RALYL expression (p=0.039), N stage (p=0.008) and distant metastasis (p<0.001) were independent prognosis factors for the overall survival of ccRCC patients. We demonstrated that the expression of RALYL was significantly low in ccRCC and correlated with a poor prognosis in a large number of clinical samples. Our findings showed that RALYL may be a potential therapeutic target as well as a poor prognostic factor.
The wasabi was planted in glasshouse to investigate the effect shade level (0, 10, 30, 70%) on growth and photosynthetic response. The net photosynthetic rate and stomatal conductance were the highest under 10% shading rate in greenhouse. The lowest transpiration rate was in the non-shading treatment. 70% shading rate significantly reduced in photosynthetic characteristics. The number of leaf was decreased with increasing of shading rate. Leaf and petiole weight was the highest 10% shading and then followed by the non-shading treatment. Number of leaves, leaf, petiole and total weight were severely declined in 70% shading treatment. Shading rate (0%, 10%, and 30%) did not significantly influenced on the leaf length, leaf width and root growth. 10% shading level was the most effective for wasabi leaf production in hydroponics.
Allergy is a multi-factorial disease influenced by genetic and environmental factors. As the number of allergy-affected people is increasing in developed countries, there is an increasing interest in genetic predisposition to the allergy. A number of genes and chromosomal region have been identified to be linked to allergy including rhinitis, asthma and atopy. In order to understand the genetic background for the allergy-affected people, we investigated genetic predisposition among students enrolled in Busan Science Academy. Among 138 students, about 30% students had some allergy-related disorder including rhinitis, asthma and atopy. We analyzed several single nucleotide polymorphisms (SNPs) within two genes, Inter-leukin-4(IL-4) and Interleukin-4 receptor(IL-4R), which are involved in the induction of allergy reaction with the Th2 immunity. For 96 samples obtained from students, we analyzed 9 SNPs including -590 C/T and -34 C/T in IL-4, and I75V, Q576R, E375A, e406R, 5411L, S761P and S727A in IL-4R. From the analysis, these SNPs showed slight differences among normal and allergy-affected students, but these differences was not enough to predict the predisposition to the allergy. In contrast to previous reports, we could not find SNP(s) related with allergy. These results suggest that genetic tests recently performed in Korea widely have to be reassessed for its validity of genetic predisposition. [Supported by grants from MOST]
In recent years the number of patients waiting for organ transplantation has greatly outpaced the supply of human organs available, which leads to a renewed interest in pig-to-human xenotransplantation as an alternative. However, one of the biggest barriers in the xenotransplantation is presence of porcine endogenous retroviruses(PERV) that can infect human cells. In this study, to present a possible solution for this problem we tried to inhibit expression of PERVs using shRNAs(short hairpin RNA) at the level of RNA synthesis and virus release. The shRNA targeting the sequence of PERV A, B type was cloned into pSIREN-RetroQ vector under the control of polymerase-III U6-RNA gene promoter. Quantitative real-time PCR was performed to detect my alterations in mRNA production of PERV A, B targeted by the shRNA in each done. Depending on the target sequence of the shRNA, the transcription of PERV was decreased to as much as 4% and the number of progeny viruses was reduced to less than 1/200,000. Transgenic pigs producing such shRNAs may result in a highly reduced PERV expression in cells and organs, which is a prerequisite for safe xenotransplantations.
Secondary prevention via earlier detection would afford the greatest chance for a cure in premalignant lesions. We investigated the exomic profiles of non-malignant and malignant changes in head and neck squamous cell carcinoma (HNSCC) and the genomic blueprint of human papillomavirus (HPV)-driven carcinogenesis in oropharyngeal squamous cell carcinoma (OPSCC). Whole-exome (WES) and whole-genome (WGS) sequencing were performed on peripheral blood and adjacent non-tumor and tumor specimens obtained from eight Korean HNSCC patients from 2013 to 2015. Next-generation sequencing yielded an average coverage of $94.3{\times}$ for WES and $35.3{\times}$ for WGS. In comparative genomic analysis of non-tumor and tumor tissue pairs, we were unable to identify common cancer-associated early mutations and copy number alterations (CNA) except in one pair. Interestingly, in this case, we observed that non-tumor tonsillar crypts adjacent to HPV-positive OPSCC appeared normal under a microscope; however, this tissue also showed weak p16 expression. WGS revealed the infection and integration of high-risk type HPV16 in this tissue as well as in the matched tumor. Furthermore, WES identified shared and tumor-specific genomic alterations for this pair. Clonal analysis enabled us to infer the process by which this transitional crypt epithelium (TrCE) evolved into a tumor; this evolution was accompanied by the subsequent accumulation of genomic alterations, including an ERBB3 mutation and large-scale CNAs, such as 3q27-qter amplification and 9p deletion. We suggest that HPV16-driven OPSCC carcinogenesis is a stepwise evolutionary process that is consistent with a multistep carcinogenesis model. Our results highlight the carcinogenic changes driven by HPV16 infection and provide a basis for the secondary prevention of OPSCC.
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