• Animal Bioscience
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• 제35권5호
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• pp.740-751
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• 2022

Objective: The study aimed to evaluate the effects of a mixture of encapsulated essential oils (EOs) addition on nutrient digestion, serum biochemical parameters, peripheral blood alpha-naphthyl acetate esterase (ANAE), and acid phosphatase (ACP-ase) positive lymphocyte ratios and intestinal morphology in laying hens. Methods: A total of 320 laying hens of 48-wk-old were randomly allotted into 4 treatment groups with 10 replicates of 8 birds in each replicate. The birds were fed a basal diet (control) or the diet added with mixture of EOs (which consist of eugenol, nerolidol, piperine, thymol, linalool, and geraniol) at 50, 100, and 200 mg/kg for period of 84 days. Results: The addition of EOs at 100 or 200 mg/kg increased the dry matter, organic matter, and crude protein digestion as compared to control. The addition of all doses of EOs did not affect serum gamma glutamyl transferase, alanine aminotransferase, and P but increased serum asparate aminotransferase (AST) concentration. The addition of 200 mg/kg EOs increased serum creatinine, while 100 mg/kg decreased Ca concentration. The addition of 100 and 200 mg/kg EOs generally improved ANAE and ACP-ase positive peripheral blood lymphocyte ratios and intestinal morphology. Conclusion: It can be concluded that, the addition of 100 or 200 mg/kg encapsulated EOs generally increased apparent nutrient digestion and serum AST concentration, improved ANAE and ACP-ase positive peripheral blood lymphocytes and intestinal morphology in laying hens.

• Clinical and Experimental Vaccine Research
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• 제12권1호
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• pp.47-59
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• 2023

Purpose: The development and study of hepatitis C virus (HCV) vaccine candidates' individualized responses are of great importance. Here we report on an HCV DNA vaccine candidate based on selected envelope (E1/E2) epitopes. Besides, we assessed its expression and processing in human peripheral blood mononuclear cells (PBMCs) and in vivo cellular response in mice. Materials and Methods: HCV E1/E2 DNA construct (EC) was designed. The antigen expression of EC was assayed in PBMCs of five HCV-uninfected donors via a real-time quantitative polymerase chain reaction. Serum samples from 20 HCV antibody-positive patients were used to detect each individual PBMCs expressed antigens via enzyme-linked immunosorbent assay. Two groups, five Swiss albino mice each, were immunized with the EC or a control construct. The absolute count of lymph nodes' CD4⁺ and CD8⁺ T-lymphocytes was assessed. Results: Donors' PBMCs showed different levels of EC expression, ranging between 0.83-2.61-fold in four donors, while donor-3 showed 34.53-fold expression. The antigens expressed in PBMCs were significantly reactive to the 20 HCV antibody repertoire (all p=0.0001). All showed comparable reactivity except for donor-3 showing the lowest reactivity level. The absolute count % of the CD4⁺ T-cell significantly increased in four of the five EC-immunized mice compared to the control group (p=0.03). No significant difference in CD8⁺ T-cells % was observed (p=0.89). Conclusion: The inter-individual variation in antigen expression and processing dominance was evident, showing independence in individuals' antigen expression and reactivity levels to antibodies. The described vaccine candidate might result in a promising natural immune response with a possibility of CD4⁺ T-cell early priming.

• Preventive Nutrition and Food Science
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• 제16권3호
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• pp.248-252
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• 2011

Soy protein based formula (SPF) has been developed for infants who are at a high risk for atopic dermatitis (AD) and cow's milk protein allergy (CMA). We performed this study to evaluate the therapeutic efficacy and safety of SPF compared to conventional hydrolyzed cow's milk formula (hCMF) in the feeding of infants with AD and CMA. 38 infants (12 to 24 months of age) diagnosed with CMA and AD were randomized to receive either SPF or hCMF for 12 weeks. Follow-up was conducted at 4, 8 and 12 weeks. Growth parameters of the infants were evaluated during each visit. Clinical evaluations, including AD severity scores, pruritus, specific immunoglobulin E (IgE) (cow's milk protein and soy protein) levels of peripheral blood, were made at enrollment and week 12. Analysis was performed on the 32 infants (SPF: n=16, hCMF: n=16) who completed the 12-week intervention. Eczema area and severity index (EASI) scores, a measure of the severity of AD, and pruritus were significantly reduced after 12 weeks compared to enrollment in the both groups; however, the median changes for EASI scores and pruritus were not statistically different between the two groups. The growth parameters did not differ significantly between both groups at any assessed time point. This study suggests that SPF could be useful in decreasing the severity of AD without affecting infant growth status. Therefore SPF could provide an adequate and safe alternative to hCMF in treating infants with AD and CMA during the first 12 to 24 months of their life.

• Tuberculosis and Respiratory Diseases
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• 제42권5호
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• pp.703-712
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• 1995

연구배경: 말초혈액 단핵구는 정상적인 면역반응에 중요한 세포로서 탐식 기능과 항원 제공 기능외에 다양한 종류의 Cytokine을 생성하며, 이중 IL-8은 호중구에 대한 강력한 주화 작용과 직접적인 활성화 작용 등을 통해서 염증 작용을 증폭시키는 역할을 하는 것으로 알려져 었다. 따라서 IL-8 작용의 억제를 통하여 항염 효과를 기대할 수 있으며, 이러한 작용을 나타낼 것으로 기대되는 Dexamethasone, $PGE_2$, Indomethacin 및 IFN-$\gamma$를 이용하여 이러한 약제들이 말초혈액 단핵구에서 내독소 자극에 의한 IL-8의 생성 자극 효과에 어떤 영향을 미치는 지에 대해서 알아보았다. 방법: 말초혈액 단핵구는 자원자에게서 채취한 혈액을 Ficoll Hypaque density gradient method에 의해서 백혈구 층을 분리한 뒤에 플라스틱 접시에 부착함으로써 분리하였다. Dexamethasone, $PGE_2$, Indomethacin은 내독소로 자극하기 한 시간 전, 후에 처치하였고, IFN-$\gamma$는 내독소로 자극하기 한 시간 전에만 전처치 하였다. 내독소로 자극한 뒤 4시간 뒤에 RNA를 extraction한 뒤 Northern blot analysis를 시행하여 IL-8 mRNA를 분석하였고, 24시간 뒤에 ELISA를 통하여 IL-8 단백을 정량하였다. 이 과정을 3회 반복하여 다음과 같은 결과를 얻었다. 결과: 1) Dexamethasone의 전, 후 처치는 내독소 자극에 의한 IL-8 mRNA와 단백의 생성을 감소시켰다. 2) IFN-$\gamma$의 단독 투여는 내독소 자극을 받지 않은 군에서 IL-8 mRNA와 단백의 생성을 감소시켰다. 3) 내독소 투여 전에 IFN-$\gamma$를 전 처치 하였을 때, 내독소 단독 투여군에 비해 IL-8 mRNA 발현은 감소하였으나 IL-8 단백의 분비에는 차이가 없었다. 4) $PGE_2$와 Indomethacin은 각각 사용한 농도($10^{-6}M$, $10{\mu}M$)에서는 내독소 자극에 의한 IL-8 mRNA 및 단백의 생성에 별다른 영향을 미치지 못하였다. 결론: Dexamethasone은 말초혈액 단핵구에서 내독소의 자극에 의한 IL-8 mRNA 발현 및 단백의 생성을 억제함으로써 염증 반응을 억제할 것으로 기대된다.

• IMMUNE NETWORK
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• 제1권3호
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• pp.250-259
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• 2001

To determine if initial infection with Mycobacterium tuberculosis changes the balance of cytokines between T cells and macrophages, we evaluated interferon (IFN)-${\gamma}$), interleukin-12 (IL)-12, and tumor necrosis factor (TNF)-${\alpha}$ productions by peripheral blood mononuclear cells (PBMC) from 15 untreated active pulmonary tuberculosis (TB) patients and 12 healthy tuberculin reactors (HTR). Freshly isolated PBMC were stimulated with Triton X-100 solubilized protein (TSP), 30-kDa or purified protein derivatives (PPD) antigen for 6, 18 and 96 hours. IL-12 p40 production by antigen-stimulated PBMC from TB patients was significantly decreased compared with that in HTR. In addition, IFN-${\gamma}$ production was significantly depressed in TB patients than that in HTR at a 96-hr stimulation. However, TNF-${\alpha}$ production was significantly higher in antigen-stimulated PBMC from TB than that of HTR. A pronounced increase in IFN-${\gamma}$ protein followed neutralization of IL-10 in early TB patients. However, neutralization of TNF-${\alpha}$ did not significantly alter IFN-${\gamma}$ induction in PBMC from TB patients. There were no significantly differences in the cytokine productions among three proteins, TSP, 30-kDa or PPD antigen. These results indicate that development of TB may be strongly associated with dysregulated productions of IL-12, IFN-${\gamma}$ and TNF-${\alpha}$, during the initial immune responses to M. tuberculosis. Further understanding of operative cytokine networks during human immune cell responses to protein antigens of M. tuberculosis may improve strategies for vaccine development.

• Animal cells and systems
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• 제8권2호
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• pp.125-133
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• 2004

We have extended our previous work that cross-linking CD4 molecules using specific MAb induced antigen nonspecific, MHC unrestricted killing of virally infected target cells by CD$4^+$We have extended our previous work that cross-linking CD$4^+$ molecules using specific MAb induced antigen nonspecific, MHC unrestricted killing of virally infected target cells by CD$4^+$ T cells. The killing activity of antibody activated CD$4^+$T cells was completely blocked by herbimycin A, a protein tyrosine kinase (PTK) inhibitor, but not by bisindolylamaleimide, a protein kinase C (PKC) inhibitor. Herbimycin A treated human or bovine peripheral blood CD$4^+$T cells lacked PTK activity and failed to kill virally infected target cells even after cross-linking of CD4 molecules. The CD$4^+$cross-linking failed to induce effector cell proliferation or the transcription of TNF${\beta}$ Upregulation of TNF${\beta}$ was induced by incubating the antibody activated effector cells with BHV-1 infected D17 target cells for 10 h. Anti-TNF${\beta}$ antibody partially abolished (13-44%) the direct effector cell-mediated antiviral cytotoxicity. However, this antibody neutralized 70 to 100% of antiviral activity of effector and target cell culture supernatants against BHV-1 infected D17 cells. The inhibition level of the antiviral activity by the antibody was dependent on the effector and target cell ratio. These results support the hypothesis that increased p$56^ICK enzyme activity in effector cells transduces a signal critical for effector cell recognition of viral glycoproteins expressed on the target cells. Following target cell recognition, lytic cytokines known to participate in target cell killing were produced. A better understanding of the killing activity displayed by CD$4^+$T lymphocytes following surface receptor cross-linking will provide insight into the mechanisms of cytotoxic activity directed toward virally-infected cells.T cells. The killing activity of antibody activated CD$4^+$T cells was completely blocked by herbimycin A, a protein tyrosine kinase (PTK) inhibitor, but not by bisindolylamaleimide, a protein kinase C (PKC) inhibitor. Herbimycin A treated human or bovine peripheral blood CD4T cells lacked PTK activity and failed to kill virally infected target cells even after cross-linking of CD4molecules. The CD4 cross-linking failed to induce effector cell proliferation or the transcription of TNF$\beta$. Upregulation of TNF$\beta$ was induced by incubating the antibody activated effector cells with BHV-1 infected D17 target cells for 10 h. Anti-TNF$\beta$ antibody partially abolished (13-44%) the direct effector cell-mediated antiviral cytotoxicity. However, this antibody neutralized 70 to 100% of antiviral activity of effector and target cell culture supernatants against BHV-1 infected D17 cells. The inhibition level of the antiviral activity by the antibody was dependent on the effector and target cell ratio. These results support the hypothesis that increased $56^ICK enzyme activity in effector cells transduces a signal critical for effector cell recognition of viral glycoproteins expressed on the target cells. Following target cell recognition, lytic cytokines known to participate in target cell killing were produced. A better understanding of the killing activity displayed by CD$4^+$T lymphocytes following surface receptor cross-linking will provide insight into the mechanisms of cytotoxic activity directed toward virally-infected cells.

• Applied Microscopy
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• 제9권1호
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• pp.57-69
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• 1979

과피가 녹색이고 배의 길이가 $250{\mu}$인 미성숙 과정을 거쳐 발아시까지 배 및 배유세포의 미세구조 변화를 관찰한 결과를 요약하면 다음과 같다. 1. 배의 길이가 $250{\mu}$인 미성숙 종자의 배세포는 세포질이 비교적 충실하나 mitochondria의 Cristae 및 plastid는 미분화상태이고 가끔 dictyosome이 관찰되었다. 액포안에는 전자밀도가 높은 globoid 를 함유하는 것도 있었다. 소포체는 대부분 한면소포체이고 외형질에는 spherosome을 많이 함유하고 있었다. 배유세포는 spherosome과 이들로 둘러싸인 전자밀도가 높고 균질성인 protein body로 충만되었고 이들로 인하여 세포기관은 관찰되지 않았다. 2. 과피가 홍색인 종자의 배세포는 cristae가 발달한 mitochondria가 관찰되었으며, 전자밀도가 높은 globoid가 증가하였고 액포는 확장되었다. 배유세포에는 spherosome이 증가하고 있었으며 protein body는 확장되어. 그 내부에는 전자밀도가 높은 globoidal crystal이 분산되어 있었다. 3. 종피가 열개한 종자의 배축의 전형성층과 유아세포에서는 Golgi vacuole과 vesicle이 발달되었으며 mitochondria의 cristae는 매우 분화되었다. spherosome은 매우 많이 나타났고 유근세포, 배축의 주변세포 및 자엽세포의 액포는 매우 분화되었다. 배유세포는 spherosome으로 충만되어 있고 그 내질에는 전자밀도가 높고 작은 입자를 지니고 있었다. Protein body는 단일막으로 둘러싸여 있음이 확인되었다. 세포질내에는 구형체와 spherosome 주변의 세포질에 acid phosphatase의 활성을 나타냈다. 4. 파종시 자엽의 외면부 세포는 spherosome이 급격히 증가하였고 protein body도 관찰되었다. 세포기관들은 분화되어 있었고 전분을 함유하는 색소체가 출현하였다. 5. 파종시킨 후 발아할 때까지 자엽의 외측 $2{\sim}3$열세포, 배축의 주변세포, 유근부 세포에서는 spherosome이 증가하였고 전분립을 함유하는 plastid 도 증가하였다. 배를 구성하는 세포의 내형질의 액포부근에는 mitochondria와 microbody 가 혼재하고 있음이 관찰되었다. 발아시기가 가까워 질수록 배와 연접하는 배유세포에는 액포가 매우 분화되고 spherosome은 감소되는 반면 액포내에는 무정형의 전자밀도가 높은 물질이 증가하였다. 그밖의 배유세포에는 spherosome이 감소됨에 따라 mitochondria는 증가되고 전자밀도가 높은 소구체가 형성되어 증가되었다. Protein body의 외질은 감소되고 전자밀도가 낮은 부위는 확장되며 이는 서로 융합하여 외연부는 없어지고 기질속에 소입자들이 형성되었다. 6. 발아기의 배유세포안에는 spherosome이 감소되고, protein body는 서로 융합되어 한 세포내에 $2{\sim}3$개로 충만되어 있었다.

• Tuberculosis and Respiratory Diseases
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• 제39권4호
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• pp.334-342
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• 1992

연구배경 : Mycobacteria 감염에 의한 폐결핵은 세포매개성 면역 반응이 관계한다고 알려져 있는 바, 먼저 Mycobacteria가 흡입되면 T 임파구가 활성화되어 세포매개성 면역 반응을 나타낸다. 결핵환자에서 tuberculin-purified protein derivative에 대한 피부 면역반응은 주로 세포매개성 면역반응에 의해 이루어짐으로 활동성 폐결핵 환자에서는 양성반응을 나타내어야하나 활동성 폐결핵 환자에서도 음성반응(anergy)을 나타내는 경우가 있다. 저자들은 폐결핵 환자에서 말초혈액 및 기관지 폐포세액내의 임파구의 세포조성과 이들이 피부반응에 미치는 영향을 조사함으로써 활동성 폐결핵 환자에서의 PPD에 대한 피부반응 검사상 음성으로 나타나는 이유를 알아보고자 하였다. 방법 : 11명의 정상대조군, 20명의 활동성 폐결핵 환자를 대상으로 PPD에 대한 피부 반응 검사 및 말초 혈액 및 기관지 폐포세척액의 임파구의 아형을 분석하였다. 결과 : 1) 기관지 폐포세척액내의 임파구는 환자군에서 정상대조군에 비하여 유의하게 증가되어 있었고($25.2{\pm}4.8$ vs $6.6{\pm}1.3%$, p<0.01), 단구세포(monocyte)는 환자군에서 정상 대조군에 비하여 유의하게 감소되어 있었다($69.6{\pm}5.7$ vs $89.2{\pm}1.4%$, p<0.05). 2) PPD에 대한 양성 반응을 보인 환자군과 음성 반응을 보인 환자군 사이의 기관지 폐포세척 세포 조성의 비는 차이가 없었다. 3) 환자군에서 기관지 폐포세척액 내의 $CD3^+$, $CD4^+$ 임파구의 조성비는 말초혈액보다 유의하게 증가되어 있었고($CD3^+$: $76.56{\pm}2.18$ vs $57.59{\pm}2.17%$, p<0.001; $CD4^+$: $51.24{\pm}2.33$ vs $35.20{\pm}2.32%$, p<0.005), $CD3^+IL-2R^+$, $CD3^+HLA-DR^+$ 임파구의 조성비도 말초혈액보다 증가되어 있었다($CD3^+IL-2R^+$:$2.41{\pm}0.57$ vs $0.93{\pm}0.16%$, p<0.005; $CD3^+$ HLA-$DR^+$: $16.92{\pm}3.89$ vs $3.94{\pm}0.70%$, p<0.005). 4) 환자군중 PPD에 대한 양성반응을 보인 군과 음성반응을 보인 군 사이에는 기관지 폐포세척액내의 임파구의 조성의 차이 및 수에서 차이가 없었다. 5) 환자군에서 PPD에 대한 피부반응과 기관지 폐포세척 세포의 조성비 사이에는 모두 상관관계가 없었으며, PPD에 대한 피부반응과 기관지 폐포세척액 및 말초 혈액내의 임파구아형의 조성과도 상관관계가 없었다. 결론 : 결론적으로 기관지 폐포세척액내의 염증 세포와 세포매개성 면역 반응과는 직접적인 관계가 없음을 알수 있었으며, 활동성 폐결핵 환자에서의 지연성 피부 반응의 저하는 임파구의 구획에 의한 결과라는 것을 배제할 수 없었다.

• Nutrition Research and Practice
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• 제8권4호
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• pp.368-376
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• 2014

BACKGROUND/OBJECTIVES: The objectives of this study were to investigate the effects of lycopene on the migration, adhesion, tube formation capacity, and p38 mitogen-activated protein kinase (p38 MAPK) activity of endothelial progenitor cells (EPCs) cultivated with high glucose (HG) and as well as explore the mechanism behind the protective effects of lycopene on peripheral blood EPCs. MATERIALS/METHODS: Mononuclear cells were isolated from human peripheral blood by Ficoll density gradient centrifugation. EPCs were identified after induction of cellular differentiation. Third generation EPCs were incubated with HG (33 mmol/L) or 10, 30, and $50{\mu}g/mL$ of lycopene plus HG. MTT assay and flow cytometry were performed to assess proliferation and apoptosis of EPCs. EPC migration was assessed by MTT assay with a modified boyden chamber. Adhesion assay was performed by replating EPCs on fibronectin-coated dishes, after which adherent cells were counted. In vitro vasculogenesis activity was assayed by Madrigal network formation assay. Western blotting was performed to analyze protein expression of both phosphorylated and non-phosphorylated p38 MAPK. RESULTS: The proliferation, migration, adhesion, and in vitro vasculogenesis capacity of EPCs treated with 10, 30, and $50{\mu}g/mL$ of lycopene plus HG were all significantly higher comapred to the HG group (P < 0.05). Rates of apoptosis were also significantly lower than that of the HG group. Moreover, lycopene blocked phosphorylation of p38 MAPK in EPCs (P < 0.05). To confirm the causal relationship between MAPK inhibition and the protective effects of lycopene against HG-induced cellular injury, we treated cells with SB203580, a phosphorylation inhibitor. The inhibitor significantly inhibited HG-induced EPC injury. CONCLUSIONS: Lycopene promotes proliferation, migration, adhesion, and in vitro vasculogenesis capacity as well as reduces apoptosis of EPCs. Further, the underlying molecular mechanism of the protective effects of lycopene against HG-induced EPC injury may involve the p38 MAPK signal transduction pathway. Specifically, lycopene was shown to inhibit HG-induced EPC injury by inhibiting p38 MAPKs.

• Journal of Nutrition and Health
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• 제39권4호
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• pp.323-330
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• 2006

Peripheral insulin resistance in obese/type II diabetes animals results from an impairment of insulin-stimulated glucose uptake into skeletal muscle. Insulin stimulate the translocation of GLUT4 from intracellular location to the plasma membrane. Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) is implicated in mediation of fusion of GLUT4-containing vesicle with the plasma membrane. Present study investigated regulatory effects of Rhodiola sachalinensis administration and exercise training on the expression of GLUT4 protein and SNAREs protein in skeletal muscles of obese Zucker rats. Experimental animals were randomly assigned into one of five groups ; lean control(LN), obese control(OB), exercise-treated(EXE), Rhodiola sachalinensis-treated(Rho), combine of Rho & EXE (Rho-EXE). All animals of exercise training (EXE, Rho-EXE) performed treadmill running for 8 weeks, and animals of Rho groups (Rho, Rho-EXE) were dosed daily by gastric gavage during the same period. After experiment, blood were taken for analyses of glucose, insulin, and lipids levels. Mitochondrial oxidative enzyme (citrate synthase, CS ; $\beta$-hydroxyacyl-CoA dehydrogenase, $\beta$-HAD) activity were analysed. Skeletal muscles were dissected out for analyses of proteins (GLUT4, VAMP2, syntaxin4, SNAP23). Results are as follows. Exercise and/or Rhodiola sachalinensis administration significantly reduced body weight and improved blood lipids (TG, FFA), and increased insulin sensitivity. Endurance exercise significantly increased the activity of mitochondrial enzymes and the expression of GLUT4 protein, however, administration of Rhodiola sachalinensis did not affect them. The effect of exercise and/or Rhodiola sachalinensis administration on the expression of SNARE proteins was unclear. Our study suggested that improvement insulin sensitivity by exercise and/or Rhodiola sachalinensis administration in obese Zucker rats is independent of expression of SNARE proteins.