• Molecular & Cellular Toxicology
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• v.3 no.4
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• pp.299-305
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• 2007

The endocrine disrupting chemical, di (2-ethylhexyl) phthalate (DEHP) is a plasticizer used in polyvinyl chloride products ubiquitous in our daily lives. DEHP has potentially adverse effects on the liver, kidney, lung, heart, reproductive organs and endocrine systems. Many toxicological data on the DEHP toxicity have been stated, but complete protein profiles have not yet been reported. In this study, DEHP-induced oxidative DNA damage in rat lymphocyte was evaluated by Comet assay (single-cell gel electrophoresis) for the first time. Moreover, DEHP-induced protein profile alterations were examined in rat liver by using toxicoproteomic tools. 34 protein spots in the liver were identified to be significantly deregulated by DEHP on the 2-dimensional gel. Among them, 20 spots were up-regulated and 14 spots down-regulated by DEHP.

• Herbal Formula Science
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• v.25 no.3
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• pp.349-362
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• 2017

Objectives : We investigated whether the components of Ailanthus altissima induced apoptotic cell death in Jurkat acute lymphoblastic leukemia (ALL) cells. Methods : Regulation of cell proliferation is a complex process involving the regulated expression and/or modification of discrete gene products, which control transition between different stages of the cell cycle. Results : Upon treatments with Ailanthus altissima, the concentration-dependent inhibitions of cell viability were observed as compared to untreated control group. The capability of Ailanthus altissima to induce apoptosis was associated with proteolytic cleavage of specific target proteins such as poly(ADP-ribose)polymerase (PARP) and beta-catenin proteins suggesting the possible involvement of caspases. Ailanthus altissima also caused apoptosis as measured by cell morphology and DNA fragmentation. Conclusions : These results indicate that the increase of apoptotic cell death by Ailanthus altissima may be due to the inhibition of cell cycle in human Jurkat lymphocytes. Conclusively, these current and further findings will provide novel approaches to understanding and treating major diseases.

• Reproductive and Developmental Biology
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• v.29 no.2
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• pp.109-115
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• 2005

Hematopoietic stem cells (HSC) are multipotent cells that reside in the bone marrow and replenish all adult hematopoietic lineages throughoutthe lifetime. In this study, we analyzed the expression of receptors of $P2Y_{10}$, purinergic receptor families in murine hematopoietic stem cells, hematopoietic progenitor cells. In addition, the biological activity of $P2Y_{10}$ was investigated with B lymphocyte cell line, Ba/F3 in effect to cell growth and cell cycle. From the analysis of expression in hematopoieticstem cell. and progenitor with RT-PCR, $P2Y_{10}$ was strongly expressed in murine hematopoieticstem cells (c-kit+ Sca-l+ Lin-) and progenitor cell population, such as c-kit- Sca-l+ Lin-, c-kit+ Sca-l- Lin- and c-kit- Sca-l- Lin-. To investigate the biological effects by $P2Y_{10}$, retroviral vector from subcloned murine $P2Y_{10}$ cDNA was used fur gene introduction into Ba/F3 cells, and stable transfectant cells were obtained by flow cytometry sorting. In cell proliferation assay, the proliferation ability of $P2Y_{10}$ receptor gene­transfected cells was strongly inhibited, and the cell cycle was arrested at G1 phase. These result suggest that the $P2Y_{10}$ may be involved the biological activity in hematopoietic stem cells and immature B lymphocytes.

• BMB Reports
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• v.35 no.3
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• pp.262-266
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• 2002

A non-isotopic neutravidin-based reverse transcriptase (RT) assay adapted for high throughput screening of HIV activity is described. Using a 96-well microtitre plate, HIV particles are lysed and the RT enzyme released into a reaction mixture containing poly(A) RNA, biotinylated oligo d(T) and fluorescein-labelled dUTP (FI-dUTP). With poly(A) as a template and oligo d(T) as primer, the viron RT incorporates FI-dUTP into an elongating DNA strand. The resulting product is captured on a neutravidin-coated 96-well plate and the unincorporated nucleotides removed by a series of washing steps. A simple ELISA is subsequently performed using a monoclonal antifluorescein antibody conjugated to alkaline phosphatase. Quantification of RT activity is facilitated by a colorimetric readout. The assay was validated in the context of a diagnostic HIV-1 phenotyping assay. Using supernatants from HIV-1 infected lymphocyte cultures the assay was shown to be as sensitive as a radioactive assay and the RT activity correlated well with levels of cell-asociated HIV-p24. Importantly, even minor reductions of RT activity by virus variants with reduced fitness could be distinguished.

• BMB Reports
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• v.44 no.6
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• pp.381-386
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• 2011

In the present study, we characterized the function of HS1-binding protein 3 (HS1BP3), which is mutated in essential tremor and may be involved in lymphocyte activation. We found that HS1BP3 localized to the mitochondria and endoplasmic reticulum partially. Overexpression of HS1BP3 induced apoptosis in HEK293T and HeLa cell lines. When these cell lines were transfected with HS1BP3, they exhibited nuclear DNA condensation, externalization of phosphatidylserine (PS), and cleavage of poly ADP ribose polymerase (PARP). Furthermore, suppression of HS1BP3 or HS1 expression attenuates HS1BP3 induced apoptosis. In addition, HS1BP3 enhanced activator protein 1 (AP-1)-mediated transcription in a dose-dependent manner. Therefore, we conclude that HS1BP3 regulates apoptosis via HS1 and stimulates AP-1-mediated transcription.

• IMMUNE NETWORK
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• v.13 no.1
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• pp.16-24
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• 2013

CTLA-4Ig is regarded as an inhibitory agent of the T cell proliferation via blocking the costimulatory signal which is essential for full T cell activation. To improve applicability, we developed the CTLA-4Ig-CTKC in which the c-terminal lysine had been replaced by cysteine through single amino acid change. The single amino acid mutation of c-terminus of CTLA-4Ig was performed by PCR and was checked by in vitro transcription and translation. DNA construct of mutant form was transfected to Chinese hamster ovary (CHO) cells by electroporation. The purified proteins were confirmed by Western blot and B7-1 binding assay for their binding ability. The suppressive capacity of CTLA-4Ig-CTKC was evaluated by the mixed lymphocyte reaction (MLR) and in the allogeneic pancreatic islet transplantation model. CTLA-4Ig-CTKC maintained binding ability to B7-1 molecule and effectively inhibits T cell proliferation in MLR. In the murine allogeneic pancreatic islet transplantation, short-term treatment of CTLA-4Ig-CTKC prolonged the graft survival over 100 days. CTLA-4Ig-CTKC effectively inhibits immune response both in MLR and in allogeneic islet transplantation model, indicating that single amino acid mutation does not affect the inhibitory function of CTLA-4Ig. CTLA-4Ig-CTKC can be used in vehicle-mediated drug delivery system such as liposome conjugation.

• Korean Journal of Poultry Science
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• v.47 no.2
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• pp.95-105
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• 2020

This study presents the production characteristics and physiological characteristics of five Korean native chicken (KNC) breeds consisting of Hwanggalsaek Jaeraejong (HJ), Korean Rhode Island Red (KR), Korean White Leghorn (KL), Korean Brown Cornish (KC), and Korean Ogye (KO). We investigated their production performances, vitalities, and stress responses. We measured the survival rate, body weight, age at first egg-laying, hen-day egg production, egg weight, amount of telomeric DNA, heterophil-lymphocyte ratio (H/L ratio), and heat shock protein (HSP)-70, HSP-90α and HSP-90β gene expression levels for 493 KNCs. The survival rate was highest in KR, and lowest in KO. Body weights were steadily high in the order of KC, KR, HJ, KO and KL. Average hen-day egg production was highest in KL, and lowest in KC. While the amount of telomeric DNA was highest in KR, and lowest in KC. Furthermore, both the H/L ratio and the HSP-90β gene expression level were highest in KC, and lowest in KR. These results indicated that the KR breed was highly resistant to stress, whereas KC was more susceptible to stress. Taken together, it is considered that with improvements the KC breed would be more suited to be used as a Korean broiler breed while KL would be more appropriately used as a Korean layer breed. In addition, it is considered that the KR breed is appropriate to be used as a maternal chicken breeder based on good production capacity and excellent robustness, while the HJ breed is desirable to be improved as a high-quality Korean meat breed based on its excellent meat quality.

• Journal of Nutrition and Health
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• v.42 no.5
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• pp.442-452
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• 2009

Smoking has been known to exacerbate the initiation and propagation of oxidative stresses. Efforts have been made to reduce the smoking-induced oxidative stresses using commercial dietary supplements. Propolis is the resinous substance collected by bees from the leaf buds and bark of trees, especially poplar and conifer trees. In this trial, we examined whether a daily supplementation of 800 mg propolis can protect endogenous lymphocytic DNA damage and modulate antioxidative enzyme activities and the level of antioxidant vitamin in smokers using a placebo-controlled, doubleblinded cross-over trial. After two weeks of running-in period, 29 smokers (mean age 34.38 ${\pm}$ 1.73) received 6 tablets/day of either propolis or placebo pills for 4 weeks. After 2 weeks of washout period the subjects switched they pills for cross-over study. The degree of DNA damage (assessed by tail DNA, tail length and tail moment) was not significantly changed with propolis intake or placebo intake. Similarly, total antioxidant status (TAS) remained at the same level regardless of the treatment. Erythrocyte catalase, glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), plasma vitamin C and tocopherol level did not differ before and after propolis treatment, and did not differ between treatments. Putting all these results together, we would suggest that it is still too early to claim that propolis possess antioxidative activities.

• Journal of fish pathology
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• v.6 no.2
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• pp.111-122
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• 1993

The mechanism by which $V_H$ region gene segments is selected in B lymphocyte is not known. Moreover, evidence for both random and nonrandom expression of $V_H$ genes in matured B cells has been presented previously. In this report, the technique of in situ hybridization allowed us to analyze expressed $V_H$ gene families in normal B lymphocyte at the single cell level. The analysis of normal B cells in this study eliminated any posssible bias resulting from transformation protocols used previously and minimized limitation associated with sampling size. Therefore, an accurate measure of the functional and expressed $V_H$ gene repertoire in B lymphocyte could be made. One of the most important controls for the optimization of in situ hybridization is to establish probe concentration and washing stringency due to the degree of nucleotide sequence similarlity between different families which in some cases can be as high as 70%. When the radioactive $C{\mu}$ and $V_{H}J558$ RNA probes are tested on LPS-stimulated adult spleen cells, $2{\sim}4{\times}106cpm$/slide shows low background and reasonable frequency of specific positive cells. For the washing condition. 40~50% formamide at $54^{\circ}C$ is found to be optimum for the $C{\mu}$. $V_{H}S107$ and $V_{H}J558$ probes. The analyzed results clearly demonstrate that the level of each different $V_H$ gene family expression is dependent upon the complexity or size of that family. These findings are also extended to the level of $V_H$ gene family expression in separated bone marrow B cells depend upon the various stage of differentiation and conclude no preferential utilization of specific $V_H$ gene family. Thus, the utilization of VH gene segments in B lymphocyte of adult BALB/c mice is random and is not regulated or changed during the differentiation of B cells.

• Development and Reproduction
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• v.2 no.2
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• pp.165-171
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• 1998

p62 is a novel cytoplsmic protein that binds to SH2 domain of p56$^{lck}$, lymphocyte-specific protein tyrosine kinase, and the expression of p62 was observed in most tissues. In addition p62 interacts with various proteins including ubiquitin and atypical PKC isoform, indicating its diverse biological role in different tissues. However, little is known about functional connection between p62 and its binding proteins. In the present study, a novel cellular protein, p62 has been shown to bind to 14-3-3 $\tau$ isoform that is specific for T cells. Moreover, overexpression of p62 in T cells caused to delay onset of UV-induced apoptosis characterized by DNA fragmentation and breakdown of poly (ADP-ribose) polymerase (PARP). Lately, 14-3-3 proteins have been shown to mediate survival signal via interacting proapoptotic Bad protein in the Iymphocyte. These results suggested the presence of p62-mediated regulatory mechanism during apoptosis in T cells, in which activation-induced apoptotic signal could be interfered by p62 and 14-3-3 protein.n.