• Journal of Veterinary Clinics
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• v.16 no.1
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• pp.31-36
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• 1999

The aim of this study is to determine the phagocytosis-promoting factor(s) for feline peripheral blood polymorphonuclear cells (PMN) by culture supernatant from mono-nuclear cells (MNC) treated with egg white derivatives (EWD). The phagocytic activity of PMN was analyzed by a flow cytometry system. The EWD did not show direct effect on the phagocytic response of PMN. The phagocytic activity of PMN was enhanced by culture supernatant from MNC but not PMN treated with EWD. Therefore, it was suggested that the enhanced phagocytic activity of feline PMN could be mediated by humoral factor(s) released from MNC treated with EWD. Thus, the phagocytosis-promoting factor(s) in supernatant fraction from MNC culture treated with EWD were isolated by reverse phase high pressure liquid chromatography. The resulting supernatant fraction on 29.02 minutes of retention time showed high phagocytic activity of PMN. The molecular weight of this supernatant fraction was 16 to 18 kDa when analyzed by capillary electrophoresis. The isoelectric point was pH 5.76 when assessed by ion-exchange chromatography. These results suggest that EWD stimulates feline MNC to elaborate a phagocytosis-promoting factor, 16 to 18 kDa of molecular weight, which could be an important mediator for the enhancement of phagocytic activity of feline peripheral blood phagocytes. Further study will be needed to elucidate this phagocytic factor.

• Journal of Veterinary Clinics
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• v.29 no.3
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• pp.207-212
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• 2012

Fucoidan has been shown to enhance immune function. The objective of this study was to examine the in vitro effect of fucoidan on the chamotactic activity of canine peripheral blood polymorphonuclear cells (PMNs). The chemotactic activity of PMNs was evaluated by method of a modified Boyden chamber assay. The amount of interleukin (IL)-8 in the culture supernatants from peripheral blood mononuclear cells (PBMCs) treated with fucoidan was determined by means of ELISA. Fucoidan itself could not have chemoattract effects for PMNs. However, the chemotaxis of PMNs was remarkably enhanced by culture supernatant from PBMCs treated with fucoidan. Similarly, it was also increased by recombinant canine (rc) IL-8. These chemotactic activities of PMNs were inhibited by addition of anti-rcIL-8 polyclonal antibody (pAb). The amount of IL-8 in the culture supernatant from PBMCs was shown to increase upon treatment of fucoidan as compared with that of untreated PBMCs culture supernatant. These results suggest that fucoidan upregulates the chemotaxis of PMNs, which is mainly mediated by IL-8 released from fucoidanstimulated PBMCs.

• Journal of Veterinary Clinics
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• v.28 no.2
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• pp.183-189
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• 2011

The objective of this study was to examine the effect of fucoidan on the phagocytic capapcity and oxidative burst activity (OBA) of canine peripheral blood polymorphonuclear cells (PMNs). The phagocytic capacity and OBA of PMNs were evaluated simultaneously by using a flow cytometer. Fucoidan itself did not cause any direct effect on the phagocytic capacity and OBA of PMNs. However, the phagocytic capacity and OBA of PMNs were enhanced by the culture supernatant from PBMCs treated with fucoidan. The phagocytic capacity and OBA of PMNs were also increased by treatment with recombinant canine (rc) tumor necrosis factor (TNF)-${\alpha}$. The ability of the culture supernatant from fucoidan-treated PBMCs to stimulate the phagocytic capacity and OBA of PMNs was inhibited by addition of anti-rc TNF-${\alpha}$ polyclonal antibody (PAb) prior to the culture. The amount of TNF-${\alpha}$ in the culture supematant from PBMCs was shown to increase upon treatment of fucoidan as compared with that of vehicle-treated PBMCs culture supematant. The level of TNF-${\alpha}$ mRNA expression in PBMCs was also up-regulated by the fucoidan treatment. These results suggest that fucoidan has an immunoenhancing effect on the phagocytic capacity and OBA of canine PMNs, which is mainly mediated by TNF-${\alpha}$ released from fucoidan-stimulated PBMCs.

• Tuberculosis and Respiratory Diseases
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• v.52 no.2
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• pp.107-116
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• 2002

Background: Matrix metalloproteinases(MMP) are essential enzymes for tumor invasion and metastasis. Among the MMP family, elevated MMP-9 and stromelysin-3(STR-3) expression have been reported to be poor prognostic factors in lung cancer patients. To evaluate the possibility of a molecular diagnosis of lung cancer using peripheral blood, the mRNA expression level of MMP-9 and STR-3 was measured using a reverse transcriptase-polymerase chain reaction (RT-PCR) in patients with lung cancer. Methods : Ninety six patients(44 patients with lung cancer, 19 pulmonary infection, and 33 control) were included. To detect MMP-9 and STR-3 mRNA expression, RT-PCR was performed in peripheral blood mononuclear cells. ELISA was also used to measure the serum level of MMP-9. Results : MMP-9 was expressed more frequently in patients with a pulmonary infection(18/19, 94.7%) compared to lung cancer patients(26/44, 59.1%) or the controls (23/33, 69.7%) (p=0.018). On the other hand, STR-3 expression was observed more frequently in patients with lung cancer(37/44, 84.1%) compared to the lung infection patients(8/19, 42.1%) or control(20/33, 60.6%) (p=0.003). Among the lung cancer patients, MMP-9 was expressed more frequently when a tumor invaded the lymph nodes(17/24, 70.8%) compared to when a tumor did not(3/13, 23.1%) (p=0.005). The MMP-9 and STR-3 expression levels had no relationship with age, sex, tumor size, distant metastasis, or tumor histology. The serum MMP-9 concentration was not higher in lung cancer patients compared to patients with a pulmonary infection or the control subjects. Conclusion : STR-3 may be used as a diagnostic marker in the peripheral blood of lung cancer patients using RT-PCR. Further studies to evaluate the clinical significance of elevated STR-3 expression in lung cancer patients is recommended.

• Biomedical Science Letters
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• v.6 no.1
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• pp.65-72
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• 2000

In order to improve the early diagnosis of Johne's disease in ruminants, duplex polymerase chain reaction system for the detection of the etiologic agent of M. paratuberculosis and for the differentiation of other mycobacterial animal pathogens, such as M. bovis and M. avium, was applied. Genomic DNAs were purified from peripheral blood monocytes or milk macrophages and were used as templates in the duplex PCR. Detection of Mycobacterium spp. in the specimen was carried out by PCR using primer set specific to the mycobacterial 16S rDNA. And then, mycobacterial DNA-positive specimens were further differentiated with duplex PCR system which was composed of primer sets specific to 16S rDNA of M. avium complex and Is900 gene of M. paratuberculosis. The results were re-confirmed by Southern blot hybridization with oligonucleotide specific to the internal sequence of IS900 PCR amplicons. The applicability of this duplex PCR system was evaluated with DNAs extracted from clinical specimens of peripheral blood monocytes and milk macrophages. In summary, the duplex PCR amplification system described in this experiment is promising molecular technique for the early diagnosis of Johne's disease in ruminants.

• Journal of Animal Environmental Science
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• v.18 no.sup
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• pp.55-64
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• 2012

We investigated the effect of the clay mineral fortified with complex trace elements by specific fermentation microbes on growth performance and a quality of pork in this study. For the declared experimental animal, 80 heads of crossbred with Large White and Yorkshire were randomly splitted into a control group and a test group. Each group were assigned with 4 replicates and 10 heads of each replicate. When 0.3% of the fermentation product of the clay mineral (FCM) was added into feed stuff and fed for 9 weeks of experimental feeding period, the results of the feed intake, weight gains, pH of carcass, lightness of meat color, and brightness of the muscle semimembranosus were similar to those of the control group. However, dressing percentage, reddishness of carcass, and water holding capacity were improved, respectively, as much as 2.7%, 12%, and 10%, and reduced by 6% in cooking loss. In order to investigate the immune modulatory effect of fermented clay mineral, pheripheral blood mononuclear cells (PBMC) were isolated and Immune modulatory parameters were measured. The proliferation activity of PBMC from pigs fed the fermented clay mineral were significantly increased compared to control group pigs, and also those results were more clearly observed as activated with lipopolysaccharide and concanavaline A. The secretion of TNF-${\alpha}$ of the FCM group pigs showed an increasing tendency. Therefore it was suggested that the feeding of FCM which was high in cation metathesis and the value of infrared ray, activated the immune responses, and thus the production of the environment-friendly high quality pork without the use of antibiotics would be possible.

• Journal of Veterinary Clinics
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• v.28 no.2
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• pp.190-195
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• 2011

Nuclear factor ${\kappa}B$ (NF-${\kappa}B$) is a nuclear transcription factor that modulates the expression of inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$. trans-10, cis-12 (t10c12)-conjugated linoleic acid (CLA) participates in the inhibition of TNF-${\alpha}$ production upon lipopolysaccharide (LPS)-stimulation. However, in our previous study, t10c12-CLA enhanced the production of TNF-${\alpha}$ by LPS-unstimulated porcine peripheral blood mononuclear cells (PBMCs) and RAW 264.7 macrophages in vitro. To resolve this apparent contradiction, we hypothesized that the effect of t10c12-CLA on TNF-${\alpha}$ production depends on NF-${\kappa}B$ activation induced by LPS stimulation. To test this hypothesis, we assessed the in vitro effect of t10c12-CLA on TNF-${\alpha}$ production and NF-${\kappa}B$ p65 activity in LPS-stimulated and LPS-unstimulated porcine PBMCs. t10c12-CLA treatment resulted in increased TNF-${\alpha}$ production by LPS-unstimulated PBMCs but decreased TNF-${\alpha}$ production by LPS-stimulated PBMCs. t10c12-CLA increased the degradation of inhibitory ${\kappa}B$ ($I{\kappa}B$)-${\alpha}$ protein and activated NF-${\kappa}B$ p65 in LPS-unstimulated PBMCs, but had the opposite effect in LPS-stimulated PBMCs. Notably, t10c12-CLA enhanced NF-${\kappa}B$ p65 binding activity in LPS-unstimulated PBMCs exposed to caffeic acid phenethyl ester (CAPE), a NF-${\kappa}B$ inhibitor. Conversely, it inhibited NF-${\kappa}B$ p65 binding activity in LPS-stimulated PBMCs exposed to CAPE. These results suggest that t10c12-CLA may have different actions under different physiological conditions, and that its effect may be associated with a change in NF-${\kappa}B$ p65 activity.

• Tuberculosis and Respiratory Diseases
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• v.54 no.5
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• pp.542-550
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• 2003

Background : Synthetic glucocorticoids are widely used in many chronic inflammatory diseases because of their excellent anti-inflammatory activity. Enhancing the transcription of $I{\kappa}B$ and preventing activated NF-${\kappa}B$ from binding to ${\kappa}B$ sites are thought to be the underlying mechanisms. But these data are largely derived from in vitro studies using cell lines. In this study, after administrating a steroid to volunteers, we evaluated the effect on the NF-${\kappa}B$ system. Methods : Prednisolone(0.5mg/kg/d) was orally administered to 5 healthy volunteers for 7 days. Before and after the administration, we sampled their peripheral blood monocytes, and performed western blot analysis both with stimulation, using IL-$1{\beta}$, LPS, TNF, and without stimulation(baseline). We also performed EMSA after stimulation with LPS. Results : After ingestion of the steroid, baseline expressions of $I{\kappa}B{\alpha}$ were increased in two of the subjects, while suppressed degradations of $I{\kappa}B{\alpha}$ to stimulations were observed in all five. In addition, the binding capacity of NF-${\kappa}B$ after the administration was decreased. Conclusion : Steroid plays such roles as enhancing the transcription of $I{\kappa}B{\alpha}$, suppressing the DNA binding capacity of NF-${\kappa}B$, and suppressing the degradation of $I{\kappa}B{\alpha}$.

• Korean Journal of Veterinary Research
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• v.32 no.2
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• pp.175-179
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• 1992

To develope the methods for isolation and enumeration of lymphocyte subpopulations in pigs, we carried out the rosette-assay using sheep erythrocytes(SRBC) and Korean native goat erythrocytes(GRBC) as a target cells. To enumerate T lymphocytes, E-rosette methods were applied with RBC treated with various concentration of polymers such as Aet and Dex, singly or in combination. And to enumerate B lymphocytes, EAand EAC-rosette assay was adopted. The results were as follows; 1. E-RFR with polymer-untreated SRBC and GRBC was $32.9{\pm}7.9%$ and $31.3{\pm}9.4%$, respectively. On the other hand, RFR with 0.1M Aet plus 8% Dex treated SRBC and GRBC was increased about two-fold($67.8{\pm}7.4%$ and $69.8{\pm}8.5%$), respectively. 2. EA-RFR with SRBC and GRBC were $ 39.1{\pm}10.2%$ and $32.6{\pm}6.1%$, respectively. 3. EAC-RFR with SRBC and GRBC were $27.6{\pm}7.0%$ arld $21.0{\pm}3.2%$, respectively. These results showed that both SRBC and GRBC could be recommanded as a binding cells for rosetteassay to isolate of lymphocyte-subpopulations in pigs.

• Journal of Korean Medicine Rehabilitation
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• v.15 no.1
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• pp.67-75
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• 2005

목 적 : 면역 세포에서 주로 생성되는 사이토카인인 인터루킨-1 (interleukin-1)은 다양한 신경내분비계와 대사 기능에 영향을 미치며, 비만인의 말초혈액 단핵구에서 증가하는 경향이 있는 것으로 연구보고 되고 있다. 본 연구의 목적은 $IL-1{\alpha}$ 다형성과 사상체질이 비만과 관련이 있는지를 조사하는데 있다. 연구방법 : 본 연구에서는 현저한 체질량지수 (body mass index, BMI) 다양성을 보이는 건강한 한국 여성 182명을 대상으로 $IL-1{\alpha}$ 유전자형을 분석하였다. 결 과 : T 대립 유전자는 BMI 감소와 관련이 있었다. BMI $27-30kg/m^2$ 그룹(group III)에서 T 대립유전자의 빈도는 $25kg/m^2$ 미만의 그룹(group I)에 비하여 유의하게 감소하였다 (odds ratio, OR, 0.141; 0.25% CI, 0.04-0.54). 더욱이 태음인 여성에서 $IL-1{\alpha}$ T 대립유전자의 빈도가 normal group에 비하여 overweight group (BMI $27-29kg/m^2$)에서 명확하게 감소하였다 (OR, 0.139; 95% CI, 0.04-0.54). 결 론 : 본 연구에서 175명의 체질분석결과 $BMI{\geq}25$인 군들의 태음인의 빈도가 높았으며, BMI는 CC genotype를 가진 사람에서보다 CT+TT genotype를 가진 사람에게서 더 낮았다. 태음인에서 동형접합 또는 이형접합 T 대립유전자가 $BMI{\geq}25$인 군에서 유의하게 감소하였다. 이상의 결과는 한국 여성에서 $IL-1{\alpha}$ 유전자 조절부위에 위치한 -889C/T 다형성과 비만 및 사상체질의 관련되는 것을 의미한다.