• Reproductive and Developmental Biology
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• 제35권3호
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• pp.257-263
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• 2011

In general, the blood cell culture is a common method for animal chromosome preparation. However, every animal and its cells have unique physiological characteristics and functions. Hence, it is very difficult to find the suitable method of chromosome preparation using animal lymphocyte culture. This study was carried out to fine the suitable method of chromosome preparation using lymphocytes cultures in mammalians and aves including cattle, rat, mouse and chicken. To seek the optimal method of lymphocyte culture in each animal, $2^3$ factorial experiment was designed. The design evaluated three main effects in culture duration, kinds of mitogen supplements and colcemid exposure time with two levels within each effect. The mitotic index and the score of chromosome morphology were analyzed. In results, the suitable methods of lymphocyte culture for chromosome preparation were 72 hours culture, pokeweed mitogen(PWM) supplement and 90 minutes of colcemid exposure in cattle, 72 hours culture, PWM supplement and 50 minutes of colcemid exposure in chicken, 96 hours culture, concanavalin A supplement and 90 minutes of colcemid exposure in rat, and 72 hours culture, PWM supplement and 50 minutes of colcemid exposure in mouse, respectively. In conclusion, kinds of mitogen, culture duration and colcemid exposure time significantly affected the mitotic index and chromosome morphology, in animal lymphocyte culture. The interaction effects between/among treatment factors were also statistically significant.

• 대한수의학회지
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• 제31권1호
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• pp.49-53
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• 1991

The present study has been carried out to investigate the optimal condition on lymphocyte blastogenesis of rabbit lymphocytes, whole blood culture and microculture system in conjunction with a semiautomatic multiple sample harvester(SAMSH) was used to study the In vitro optimal condition of rabbit lymphocytes. Data were presented to show many variables that are involved in studying the phytohemagglutinin(PHA) and lipopolysaccharide(LPS) response of rabbit lymphocyte in a microculture system. Analysis indicated that the conditions for optimal PHA as measured by incorporation of $^3H$-TdR include: (1) use of RPMI-1640 as culture medium. (2) use of $6{\mu}g$ of PHA, per culture. (3) 48-hours culture period. Conditions for optimal stimulation with LPS mitogen were similar to those used for PHA.

• Archives of Pharmacal Research
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• 제17권4호
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• pp.236-239
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• 1994

The suppressive activity of isoflavonoids against lymphocyte proliferation in vitro was examined. Isoflabvonoid derivatives tested were isflavones isolated from Pueraia radix and synthesized 7-O-substituted biochanin A derivatives. The certain isoflavones such as biochanin A and 2-carbethoxybiochainin A were found to possess the suppressive activity against concanavaline A (Con A)-induced lymphocyte proliferation from mouse spleen. Against mixed lymphocyte culture reaction, biochanin A, 2-carbethoxybiochainin A, daidzein, formononetin, genistein and 7-O-isopropylbiochaninl A showed the suppressive activity at $10^{-5}$ M. However, all isoflavones tested did not show the suppressive activity against lymphocyte proliferation induced by B-cell mitogen, lipopolysaccharide (LPS). In general, isoflavones were revealed to be less active than flavones/flavonols.

• Journal of Community Nutrition
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• 제7권2호
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• pp.79-84
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• 2005

A vitamin B-6 (B-6) intake higher than the current Recommended Dietary Allowance (RDA) has been found to provide an improvement in immune system. Seven premenopausal women consumed their usual diet with the exception of foods relatively high in vitamin B-6 for a total of 27 d. After 7 d, all subjects received a multivitamin supplement containing 2mg B-6 and 4 subjects were given an additional 50mg of B-6 supplement for 20 d. Lymphocyte response to phytohemagglutinin (PHA) was measured before and after the supplementation. To determine the effect of different forms of B-6 on lymphocyte proliferation, cell culture media supplemented with pyridoxal (PL) and PLP, as well as B-6 free media, were tested. A 50mg B-6 supplement significantly increased vitamin B-6 status. There was no further enhancement on lymphocyte proliferation when subjects were taking an additional 50mg of vitamin B-6 supplement. In general, lymphocyte proliferation in media with either PLP or PL did not show any prominent difference. These [m-dings suggest that there may be no further benefits of a B-6 dose beyond twice that of the current RDA on lymphocyte proliferation. Further studies are necessary to examine the effect of the B-6 intake level on activities of enzymes involved in cellular B-6 metabolism in lymphocytes to provide substantial insight into the mechanisms underlying the role of B-6 in the lymphocyte proliferation.

• Tuberculosis and Respiratory Diseases
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• 제81권2호
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• pp.132-137
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• 2018

Background: The pathophysiology of chronic obstructive pulmonary disease (COPD) includes inflammation, oxidative stress, an imbalance of proteases and antiproteases and apoptosis which has been focused on lately. Abnormal apoptotic events have been demonstrated in both epithelial and endothelial cells, as well as in inflammatory cells including neutrophils and lymphocytes in the lungs of COPD patients. An increased propensity of activated T lymphocytes to undergo apoptosis has been observed in the peripheral blood of COPD patients. Therefore, the apoptosis of T lymphocytes without activating them was investigated in this study. Methods: Twelve control subjects, 21 stable COPD patients and 15 exacerbated COPD patients were recruited in the study. The T lymphocytes were isolated from the peripheral blood using magnetically activated cell sorting. Apoptosis of the T lymphocytes was assessed with flow cytometry using Annexin V and 7-aminoactinomycin D. Apoptosis of T lymphocytes at 24 hours after the cell culture was measured so that the T lymphocyte apoptosis among the control and the COPD patients could be compared. Results: Stable COPD patients had increased rates of $CD4^+$ T lymphocyte apoptosis at 24 hours after the cell culture, more than the $CD4^+$ T lymphocyte apoptosis which appeared in the control group, while the COPD patients with acute exacerbation had an amplified response of $CD4^+$ T lymphocyte apoptosis as well as of $CD8^+$ T lymphocyte apoptosis at 24 hours after the cell culture. Conclusion: Stable COPD patients have more apoptosis of $CD4^+$ T lymphocytes, which can be associated with the pathophysiology of COPD in stable conditions.

• 한국축산식품학회지
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• 제34권1호
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• pp.88-98
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• 2014

Total spleen lymphocytes, lymphocyte proliferation, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and interleukin-10 (IL-10) in spleen lymphocyte culture were studied in malnourished Wistar rats fed with goat milk yoghurt. Malnourished rats were created by using standard feed restriction as much as 50% of normal rats for 21 d. Goat milk yoghurt containing three types of microorganism e.g., Lactobacillus acidophilus, Sterptococcus thermophilus and Bifidobacterium longum derived from Lacto-B culture in powder form. After 21 d, the rats continued to receive restricted feeding and supplemented with goat milk yoghurt for 7 d. Total splenocytes were counted by hemocytometer. Splenocytes proliferation was expressed as stimulation index, whereas the TNF-${\alpha}$ and IL-10 of spleen lymphocyte culture were measured by ELISA technique. The total number of splenocytes and stimulation index of splenocytes in moderate malnourished and normal rats supplemented with goat milk yoghurt was not significantly different. The level of TNF-${\alpha}$ in the rat supplemented with goat milk yoghurt was lower (p<0.05) than the control group, whereas the level of IL-10 in the rat supplemented with goat milk yoghurt was higher (p<0.05) than the control group. In conclusion, goat milk yoghurt supplementation in malnourished rats could decrease TNF-${\alpha}$ as a representation of the pro-inflammatory cytokine, while it increases IL-10 as a representation of the anti-inflammatory cytokine.

• 생약학회지
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• 제42권4호
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• pp.354-360
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• 2011

In order to screen new allogeneic mixed lymphocyte culture (allo-MLR) inhibitor which is expected to be immunomodulating drug lead, we have investigated allo-MLR inhibitory activity on the marine-derived symbiotic microorganisms (1,895 strains) from the marine algae. The potent inhibitory activities (over 45% inhibition of proliferation at 10 and 2 ${\mu}g/ml$) without cytotoxicity were observed in the extracts of 46 strains. While, the significant stimulating activities (over 100% proliferation at 10 and 2 ${\mu}g/ml$) without cytotoxicity were observed in the extracts of 5 strains. In the second assay using 46 bioactive strains, 14 strains exhibited again significant allo-MLR inhibitory activity. Finally, 11 strains among the 14 strains inhibited proliferation and IFN-${\gamma}$ production of CD4+ T cells during the stimulation with specific antigen in the third assay. On the basis of above results, the marine algae is nice source for isolation of immunomodulating microorganism, and the marine algae-associated microorganism is also nice target for development of the new immunomodulating drug lead.

• Journal of Ginseng Research
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• 제32권3호
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• pp.250-256
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• 2008

Red ginseng is a medicinal herb widely used in East-Asia for a long time. Recently there have been a lot of studies about the effect of red ginseng on the immune responses. We investigated the differences between Korean red ginseng and Chinese red ginseng in the lymphocyte proliferation and cytokine production. The rats were divided into 3 groups, KRG(Korean Red Ginseng) group, CRG(Chinese Red Ginseng) group, and the Control group. Experimental groups were administered with Korean and Chinese red ginsengs for three months respectively. Then we obtained the lymphatic cells from spleen and compared the ability of KRG on the lymphocyte proliferation and the cytokine production after mitogen-stimulated culture to CRG. The proliferation of lymphocyte and level of $IL-1{\alpha}$ were significantly increased only in KRG group. There were significant increases in the level of $INF-{\gamma}$ in both KRG and CRG groups. There were no significant differences in the level of IL-2 and $TNF-{\alpha}$. These results indicate that KRG can induce infection-relevant immune responses much faster and higher than CRG. Furthermore, functional activation of CD8+ T-cell may be activated by red ginsengs.

• 대한미생물학회지
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• 제16권1호
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• pp.65-70
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• 1981

A microculture technique for the mixed lymphocyte culture test(MLC) in the sheep is described. The optimal incubation periods for the MLC are varied from 4 to 6 days depending on the sources of lymphocytes. It is found that a 2.5:1 stimulator-responder cell ratio and the responder cell concentration of $1{\times}10^5$ cells/well produce the highest($^3H$)-thymidine incorporation. Moreover cryopreservation of bovine lymphocytes results in satisfactory and reproducible MLC reaction. It is also evident that the MLC reactions of the sheep are under the control of histocompatibility matching between the stimulator cells and the responder cells. Significance of the technique as a too! for study on cell mediated immunity in sheep system, either normal or squamous cell carcinoma-bearing, are discussed.

• 대한핵의학회지
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• 제29권1호
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• pp.125-132
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• 1995

The purpose of this study was to establish mononuclear cell cultures such as lymphocytes or buffy coat for the biological dosimetry of in vitro Irradiation of the radionuclide Tc-99m in order to exclude the effect of residual doses seen in the cultures of whole blood. Biological do simetry of Tc-99m on cultured mononuclear cells at doses ranging from 0.05 to 6.00 Gy, by scoring unstable chromosomal aberrations(Ydr) observed in cultured lymphocytes, were performed using peripheral venous blood of healthy normal person. The results showed that; (1) In vitro irradiation of radioisotope in separated lymphocyte or buffy coat showed trace amount of residual doses of isotope after washing. Residual doses of isotopes are increased in proportion to exposed time and irradiated dose without difference between I-131 and Tc-99m. (2) We obtained these linear-quadratic dose response equations in lymphocyte and buffy coat culture after in vitro irradiation of Tc-99m, respectively (Ydr = 0.001949 $D^2$ +0.006279D + 0.000185; Ydr= 0.002531 $D^2$-0.003274 D+0.003488). In conclusion, the linear quadratic dose-response equation from in vitro irradiation of Tc-99m with lymphocyte and buffy coat culture was thought to be useful for assessing Tc-99m induced biological effects. And mono-nuclear cell cultures seem to be the most appropriate experimental model for the assessment of biological dosimetry of internal irradiation of radionuclides.