• International Journal of Oral Biology
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• v.35 no.1
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• pp.1-5
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• 2010

Human fibroblasts that maintain the structural integrity of connective tissues by secreting precursors of the extracellular matrix are typically cultured with serum. However, there are potential disadvantages of the use of serum including unnatural interactions between the cells and the potential for exposure to animal pathogens. To prevent the possible influence of serum on fibroblast cultures, we devised a serum-free growth method and present in vitro data that demonstrate its suitability for growing porcine fetal fibroblasts. These cells were grown under four different culture conditions: no serum (negative control), 10% fetal bovine serum (FBS, positive control), 10% knockout serum replacement (KSR) and 20% KSR in the medium. The proliferation rates and viabilities of the cells were investigated by counting the number of cells and trypan blue staining, respectively. The 10% FBS group showed the largest increase in the total number of cells ($1.09\;{\times}\;10^5\;cells/ml$). In terms of the rate of viable cells, the results from the KSR supplementation groups (20% KSR:64.7%; 10% KSR: 80.6%) were similar to those from the 10% FBS group (68.5%). Moreover, supplementation with either 10% ($3.0\;{\times}\;10^4\;cells/ml$) or 20% KSR ($4.8\;{\times}\;10^4\;cells/ml$) produced similar cell growth rates. In conclusion, although KSR supplementation produces a lower cell proliferation rate than FBS, this growth condition is more effective for obtaining an appropriate number of viable porcine fetal fibroblasts in culture. Using KSR in fibroblast culture medium is thus a viable alternative to FBS.

• IMMUNE NETWORK
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• v.15 no.3
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• pp.150-160
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• 2015

We previously reported that 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (PLAG) accelerates hematopoiesis and has an improving effect on animal disease models such as sepsis and asthma. The effects of PLAG supplementation on immune modulation were assessed in healthy men and women. The objective was to evaluate the effects of PLAG supplementation on immune regulatory functions such as activities of immune cells and cytokine production. A randomized double blind placebo-controlled trial was conducted. Seventy-five participants were assigned to one of two groups; all participants had an appropriate number of white blood cells on the testing day. The PLAG group (n=27) received oral PLAG supplements and the control group (n=22) received oral soybean oil supplements. IL-4 and IL-6 production by peripheral blood mononuclear cells (PBMC) were lower (p<0.001 and p<0.001, respectively) with PLAG than with soybean oil. However, the production of IL-2 and IFN-$\gamma$ by PBMC was unaltered with PLAG supplementation. The B cell proliferation decreased significantly in the PLAG group compared to the soybean oil control (p<0.05). The intake of PLAG in healthy adults for 4 weeks was deemed safe. These data suggest that PLAG has an immunomodulatory function that inhibits the excessive immune activity of immunological disorders such as atopic and autoimmune diseases. PLAG could improve the condition of these diseases safely as a health food supplement.

• Journal of Nutrition and Health
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• v.35 no.1
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• pp.5-14
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• 2002

Alteration in the syntesis or enhanced inactivation of nitric oxide(NO) can induce impairment of endothelial cell function. Insulin dependent diabetes mellitus(IDDM) is characterized by impaired endothelial function and vascular disease. NO is produced through L-arginine pathway To elucidate the hypothesis that the decreased production on NO in IDDM reflects vascular damage and the NO production can be manipulated by either dietary fat(7% of kg diet) or the oral supplementation with L-arginine(2g/kg bw), plasma markers for vascular endothelial damage and plasma lipid profiles were measured in streptozotocin(STZ)-induced diabetic rats. Diabetic or normal Sprague-Dawley rats were fed 6 different experimental diets for 4 weeks(SO : soybean oil, SOA: soybean oil + L-arginine supplementation, BT : beef tallow, BTA_ beef tallow + L-arginine supplementation, OV olive oil, OVA : olive oil + L-arginine supplementation). Plasma glucose, total cholesterel, HDL-cholesterol, LDL-cholesterol and triglyceride were measured. Endothelial markers, plasma von Willebrand factor(vWf), thromboxane B$_2$, and 6-keto PGF1$\alpha$ of aorta were measured by ELISA. Plasma NO production was evaluated through the measurement of nitrite by EIA. Feeding saturated fatty acid(SFA, BT) increased relative liver size(RLS) in diabetic rats compared to either polyunsatunted fatty acid(PUFA, SO) or monounsaturated fatty acid(MUFA, OV) The supplementation of L-arginine inhibited the liver and kidney enlargement in olive oil find diabetic rats. Plasma glucose was lower in diabetic animal find the olive oil compared to fed beef tallow and the supplementation L-arginine decreased it in diabetic rats find beef tallow significantly(p < 0.05). Plasma TXB$_2$ levels were increased due to diabetes and the value of beef tallow group showed highest value. Plasma vWf concentration of beef tallow group was higher value in normal rats and was elevated more in diabetes. In diabetic groups, the vWf concentration of olive oil group was lower than beef tallow or soybean oil group. The supplementation of L-arginine in diabetic rats decreased plasma TXB$_2$ and vWf levels significantly(p < 0.05). NO production was higher in normal olive oil fed rats and was tend to be decreased in diabetic rats and the supplementation of L-arginine recovered to normal value(p < 0.05), Olive oil supplemented with L-arginine tended to lower plasma total cholesterol and LDL-cholesterol after 4 week treatment. These results suggest that generalized vascular endothelial changes based on plasma TXB$_2$and vWf occurs in diabetic rats. and olive oil with L-arginine supplementation contributes to a better control of the hyperglycemia, endothelial changes and hypercholesterolemia accompanying diabetes as compared with beef tallow or soy bean oil in this rat model.

• Korean Journal of Bronchoesophagology
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• v.14 no.1
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• pp.29-33
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• 2008

Objectives : Oral pain without identifiable oral mucosa lesion is probably multifactorial origin, which include burning mouth syndrome (BMS), oral candidiasis and so on. The aim of this study was to analyze the characteristics of oral pain without identifiable oral mucosa lesion and to evaluate treatment outcome of those patients. Materials and Methods : We reviewed 50 patients without identifiable oral mucosa lesion who were complaint of oral pain. The patients were analyzed according tothe sites, associated symptoms, laboratory tests and fungus culture. The questionnaire included questions on their current diseases, smoking and alcoholic history, psychological factors, and symptoms. Results : The average age of patients was 60 years old. The most frequently involved site was tongue (92%), followed by palate, lower lip, oropharynx, and gingiva. 60% of the patients has psychological disorder as self reported. Culture for Candida was positive in 36% of patients and serum zinc deficiency was present in 60% of patients. Serum iron, vitamin B12, hemoglobin, folic acid deficiency were present in 6-2% of patients. Seventeen patients (65%) with BMS and twelve patients (66%) with oral candidiasis were improved after treatment. Conclusion : We recommend oral candida culture to oral pain patients without oral mucosa lesion. Zinc supplementation of zinc depletion patients may be helpful whereas other laboratory tests have no diagnostic values.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.2
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• pp.217-221
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• 2003

The present investigation was undertaken to study status of erythrocytic antioxidant enzymes in normal cycling and $\alpha$- tocopherol supplemented anestrus buffalo heifers. The pre-supplementation erythrocytic activities of superoxide dismutase (U/mg Hb), glutathione peroxidase (U/mg Hb) and glucose-6-phosphate dehydrogenase (U/g Hb) upregulated significantly (p<0.05) in anestrus heifers ($10.08{\pm}0.09$, $14.09{\pm}0.54$, $9.25{\pm}0.29$) when compared to normal cycling ones ($6.93{\pm}0.04$, $11.61{\pm}0.19$, $5.58{\pm}0.26$). The oral supplementation of $\alpha$-tocopherol (a) 3,000 mg per week per animal in anestrus heifers declined erythrocytic superoxide dismutase and glucose-6-phosphate dehydrogenase activities significantly (p<0.01) but led to non-significant increase in erythrocytic glutathione peroxidase activity. Results indicated that supplementation of $\alpha$-tocopherol to anestrus buffalo heifers mitigated the effects of oxidative stress to improve their antioxidant status.

• Journal of Community Nutrition
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• v.7 no.2
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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.

• Food Science and Biotechnology
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• v.18 no.6
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• pp.1528-1531
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• 2009

In this study, we investigated a dose-dependent hypolipidemic and antioxidant effects of tocotrienol-rich fraction (TRF) from grape seeds. After induction of hyperlipidemia for 4 weeks, rats were supplemented with different dose (5, 25, and 50 mg/kg BW/day) of TRF for 1 week. Oral administration of TRF (50 mg/kg BW/day) decreased the plasma triglyceride (TG, 162.6 mg/dL), total cholesterol (TC, 83.7 mg/dL), low density lipoprotein-cholesterol (LDL-C, 20.3 mg/dL), malondialdehyde contents (MDA, 3.3 nmol/dL), and atherogenic index (AI, 2.0) compare to high-fat diet group. These data suggest that TRF supplementation has significant health benefits through the modulation of physiological functions that include various atherogenic lipid profiles and antioxidative status in hyperlipidemia.

• Korean Journal of Plant Resources
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• v.24 no.3
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• pp.286-291
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• 2011

In this study we investigated effects of supplementation with ethyl acetate extracts of the brown alga Eisenia bicyclis on innate immune cells to evaluate the possibilities as an immunomoulator in exercise stress. Twenty male SD rats were divided into four groups and the treatments were as follows: A, no Eisenia bicyclis extract (EBE) (200 mg/kg) intake and maintained at rest ; B, no EBE intake and undergoing exercise ; C, EBE intake and undergoing exercise ; D, EBE intake and maintained at rest. After 5 weeks of oral supplementation, rats were undergoing intensive swimming exercises for 2 h and sacrificed to assess the effects on peritoneal macrophages, spleen cells and natural killer (NK) cells. We showed increasing effects on nitric oxide-inducible nitric oxide synthase (NO-iNOS) production by macrophages and no effects of NK tumoricidal activity and suppressive effects on spleen cell proliferation in exercise group. However, EBE supplementation suppressed NO-iNOS production by macrophages and increased NK tumoricidal activity and spleen cell proliferative response to mitogen in exercise group. Overall, these results that EBE supplementation has differential effects on innate immune response and could be useful as sports nutrition.

• Preventive Nutrition and Food Science
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• v.11 no.4
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• pp.289-297
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• 2006

This study examined the effect of hesperidin supplementation with an ethanol diet on lipid and antioxidant metabolism in rats. Male Sprague-Dawley rats were divided into two groups (n=10), and were assigned to one of two dietary categories: $E_8$, ethanol diet (50 g/L) for 8 wks; $E_8H_4$, ethanol diet for the first 4 wks and hesperidin (0.02%, w/w) supplemented ethanol diet for the last 4 wks. The plasma and hepatic lipids, hepatic cholesterol regulating enzyme activity, hepatic antioxidant enzyme activity and lipid peroxidation were determined. Supplementation with hesperidin for the last 4 wks during the 8 wks period of the ethanol diet, significantly increased the ADH activity. In conjunction with the chronic administration of ethanol, hesperidin supplementation resulted in a significant decrease in the hepatic cholesterol and triglyceride concentrations compared to the $E_8$ group. The hepatic HMG-CoA reductase and ACAT activities were significantly lower in the hesperidin-supplemented group. When comparing hepatic antioxidant enzyme activities, SOD, GSH-Px, and G6PD activities and GSH level were significantly higher in the $E_8H_4$ group than in the E8 group. Plasma TBARS levels were significantly lower in rats fed ethanol with hesperidin compared to the rats fed only ethanol; however, the hepatic TBARS levels were not significantly different between the groups. Accordingly, the additional hesperidin supplement with an ethanol diet might be effective for improving the hepatic lipid metabolism and antioxidant defense system.

• Nutritional Sciences
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• v.5 no.2
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• pp.60-67
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• 2002

Paeonia japonica var. pilosa $N_{AKAI}$, (PJ; Baek-Jak-Yak) is a medicinal plant which has been widely used as a component or blood-building decoctions. This study was performed to investigate the immunomodulative effects of PJ in mice, using in vitro and in vivo experiments. The immunomodulative effects were studied in vitro by determining the proliferation or mice splenocytes and the production of three kinds of cytokines (IL-1$\beta$, IL-6, TNF-$\alpha$) by mire peritoneal macrophages which were cultured with sequential fractions of PJ methanol extract (methanol, hexane, chloroform, ethylacetate, butanol and water). In an in vivo experiment using mice, different concentrations of PJ water extract were orally administrated every other day for two weeks. The production of cytokines (IL-1$\beta$, IL-6, TNF-$\alpha$) secreted by activated macrophages, and the proliferation of mice splenocytes, were used as indices for immunocompetence. In vitro supplementation using a hexane fraction of PJ in the range of 1 to 100 $\mu$ g/ml enhanced splenocyte proliferation by 1.8 to 12%, and by 10-15% using an aqueous fraction, compared to the control. IL-l$\beta$ production was significantly increased with the supplementation of butanol, hexane and water extracts of PJ Higher levels of IL-6 production were detected with supplementation of chloroform or water extracts. However, there were no significant differences in the production of TNF-$\alpha$ among the treated groups and the control. From the in vivo study, the highest proliferation of splenocytes was seen in the mice orally administrated with the PJ water extract at the concentration of 500 mg/kg body weight. In the case of cytosine production, IL-1-$\beta$, IL-6, and TNF-$\alpha$ released by activated peritoneal macrophages were augmented by the oral administration of a PJ water extract. These results indicate that Pl may enhance the immune function by regulating splenocyte proliferation and cytokine production capacity in mice.