• Journal of Veterinary Clinics
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• v.21 no.3
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• pp.236-242
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• 2004

1,2-benzopyrone can stimulate macrophages to increase the ability of phagocytosis. Peripheral blood polymorphonuclear cells (PMN) and macrophages destroy microbial organisms with reactive oxygen species (ROS), called oxidative burst activity (OBA). This study was undertaken to determine whether 1,2-benzopyrone affects the OBA on the phagocytic response of canine peripheral blood phagocytes. The OBA of phagocytes in the addition or absence of latex beads was analyzed by flow cytometry system using dihydrorhodamine 123 (DHR). The direct treatments of 1,2-benzopyrone have no effect on the OBA of peripheral blood mononuclear cells (PBMC), PMN and monocyte-rich cells regardless of addition of latex beads. When latex beads are added to PMN, the OBA of PMN was remarkably enhanced by culture supernatant from PBMC but not PMN treated with 1,2-benzopyrone. Similary, it was also enhanced by human recombinant (hr) $TNF-\alpha.$ However, when latex beads were not added to PMN, its OBA was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. The OBA of latex beads-phagocytized PBMC and monocyte-rich cells was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. These results strongly suggested that 1,2-benzopyrone has an immunoenhancing effect on the OBA of PMN when phagocytic response occurred only. This enhanced OBA may be mediated through active humoral substance(s), such as $TNF-\alpha,$ produced by PBMC stimulated with 1,2-benzopyrone.

• The Korean Journal of Cytopathology
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• v.4 no.2
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• pp.87-92
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• 1993

The effect of Roentgen rays on carcinoma of the cervix has long been of great interest to both radiologists and gynecologists. Since most cervical carcinomas are treated by irradiation, any additional knowledge either concerning the radiosensitivity of cervical tumors or their ultimate prognosis would be of value. The vaginal smear is considered to be one of convenient and rapid methods to study the effects of radiation on cervical malignancy. We observed morphologic changes in 297 cytologic preparations obtained from 60 patients who had underwent irradiation for cancer of the cervix. With the morphologic parameters such as cytoplasmic vacuolization, cytoplasmic basophilia, multinucleated giant cell formation, polymorphonuclear leucocytes (PMNL) sticking and postradiation dysplasia, we analyzed the findings in relation to the follow up time interval. The most common effect was the cytoplasmic vacuolization with basophilia of basaloid cells, which were noted in more than 90% of followed patients. The multinucleated giant ceil formation and PMNL stickering were noted in 38 cases(63%) and 48 cases(80% ) respectively. The differential diagnosis of postradiation dysplasia from recurrent or persistent carcinoma, reparative atypical cells, and regressing tumor cells was difficult and further study seems to be needed to clarify the more accurate morphologic features and biologic behavior.

• 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.15 no.1
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• pp.110-115
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• 1998

인삼 PD saponin(GPD)으로 배양한 고양이 말초혈액 단핵구세포(afNC) 배양상충 액에서 말초혈액 다형핵백혈구(PMNC)에 대한 interleukin(IL) 8 양 유주활성에 잔하여 검토 하였다. PMNC의 유주활성은 Hoyden chamber 변법으로 측정하였다. GPD를 첨가하여 배양 한 MNC배양상층액중에는 rMNC에 대한유주활성이 인정되었다. PMNC에 대하여 GPD 로 배양한 MNC 배양상층액중에 존재하는 유주활성이 IL 8 양 물질인지를 알아보기 위해 human recombinant IL 8을 이용하여 고양이 PMNC에 대해 유주팔성을 측정한 결과, GPD 로 배양한 MNC 배양상층액의 경우와 동등한 활성이 나타났다. Human IL 8 mAb를 사용하 여 GPD로 배양한 MNC 배양상층액중의 유주활성에 대한 중화반응을 살펴본 결과, GPD로 배양잔 MNC 배양상충액 및 human IL 8에 의해 증가되었던 PMNC의 유주활성은 IL 8 cAb의 첨가농토가 증가함에 따라 활성이 완전히 억제되었다. 또한 GPD로 배양한 고양이 MNC 배양상충액중의 유주활성은 열처리(4, 30, 37, 60 및 $100{\circ}C$) 및 산(pH 3.0)과 알카리 (pH 9.0)처리에도 안정성을 보여 human IL 8의 물리화학적 성상과 매우 유사하였다. 따라서 GPD로 배양한 MNC 배양상충액중에 존재하는 고양이 PMNC에 대한 유주활성은 feline IL 8 양 물질임을 강하게 시사하였다.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.2
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• pp.183-189
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• 1999

This investigation is to determine whether the surface complexation of iron influence acute pulmonary responses induced by silica. For this study, three varieties of cation complexed silica were used: $silica-H^+,\;-Zn^{2+},\;and\;-Fe^{3+},$ since the first two are not active in the transport of electrons and generate little free radicals relative to the dust with the surface iron. Rats (270 to 280 g) were intratracheally (IT) instilled with saline, $silica-H^+,\;-Zn^{2+},\;or\;-Fe^{3+}$(5 mg in 0.5 ml saline). After 4 h, cell number, type, and differentiation were analysed in the bronchoalveolar lavage cells, and the levels of lactate dehydrogenase (LDH) and total protein were determined in the lavage fluid. In addition, bronchoalveolar lavage cells were cultured, and nitric oxide production was measured using nitrate assay. Inducible nitric oxide synthase (iNOS) mRNA in the bronchoalveolar lavage cells was also determined by northern blot analysis. Differential counts of the lavage cells showed that red blood cells were increased by 9-, 8-, and 13-fold and total leukocytes (lymphocytes plus polymorphonuclear neutrophils) by 48-, 36-, and 33-fold, following IT $silica-H^+,\;-Zn^{2+},\;and\;-Fe^{3+},$ respectively compared with the saline group. Meanwhile, there were no significant differences in red blood cells and total leukocytes among any of the cation complexed silica groups. The levels of LDH and total protein in the lavage fluid were significantly increased by 3- to 4-fold. However, compared among these silica groups, $Fe^{3+}$? complexation did not significantly change the LDH activity and total protein. NO production in cultured bronchoalveolar lavage cells was elevated by 2-fold, following IT any of the silica treatments compared with the saline group. Furthermore, the steady-state levels of iNOS mRNA in the lavage cells were greatly increased. There were any differences in iNOS mRNA expression among the silica-treated groups as with NO production. These findings suggest that surface complexed iron may not influence the acute pulmonary responses resulted from 4h exposure to silica.

• Biomolecules & Therapeutics
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• v.1 no.2
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• pp.131-136
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• 1993

In the present study, the effect of $HgCl_2$on the function of human peripheral polymorphonuclear leukocytes(PMNs) was examined. PMNs were isolated from human peripheral blood with density centrifugation in Ficoll-Paque. The cells were then incubated with $0.5{\sim}5{\mu}M\;HgCl_2$and glass adherence, chemotactic activity and erythrocyte-antibody rosette forming activity were measured. $HgCl_2$ decreased the function of PMNs in all three aspects tested. $HgCl_2$significantly diminished glass adherence(40.5 {\mu}M: 92{\pm}12%$ (percentage of control, $mean{\pm}$ S.D.); 41 {\mu}M: 46{\pm}11%,$ P<0.01; $3{\mu}M: 35{\pm}7%,$P<0.01;$5{\mu}M:49{\pm}10%,$ P<0.01). Similarly, significant differences were observed in chemotactic activity after $HgCl_2$treatment compared with control (control: $0.95{\pm}0.14mm; 0.5 {\mu}M: 0.91{\pm}0.11 mm; 1 {\mu}M: 0.77{\pm}0.16mm, P<0.05; 3{\mu}M: 0.61{\pm} 0.06mm, P<0.01; 5{\mu}M: 0.15{\pm}0.03 mm, P<0.01).$ Also, 4HgCl_2$decreased the percentage of rosette-forming PMNs, indicating diminished phagocytic activity of PMNs upon $HgCl_2$ exposure compared with control (control: $58{\pm}4%; 1{\mu}M: 53{\pm}4%, p<0.05; 3{\mu}M: 49{\pm}3%, P<0.01; 5{\mu}M: 46{\pm}3%, P<0.01).$ Cell viability was not antered after $HgCl_2$treatment (483{\pm}5%$ viability in control PMNs versus $81{\pm}8%$ viability in $5{\mu}M$ Hg-treated PMNs), suggesting that the impaired PMN function after $HgCl_2$treatment was not due to nonspecific cytotoxicity induced by $HgCl_2$. $HgCl_2$-induced decrease in the function of PMNs may have some implications in depressed host susceptibilityupon bacterial challenge after mercury exposure.

• BMB Reports
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• v.35 no.5
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• pp.482-487
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• 2002

Early activation of human peripheral blood polymorphonuclear neutrophils is characterized by their morphological changes from spherical to polarized shapes. The endotoxins from enteric pathogens (S. dysenteriae type 1, V. cholerae Inaba 569B, S. typhimurium, and K. pneumoniae) were assessed by their ability to induce morphological polarization of the neutrophils as measures of early activation. Phagocytic activity, adhesion, chemokinetic locomotion, and nitroblue tetrazolium (NBT) dye-reduction ability measured the later activation of the cells. Neutrophils showed distinct morphological polarization in suspension over a wide range of concentrations of these endotoxins when were compared with those that were induced by the standard chemotactic factor, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP). It was discovered that all of the endotoxins induced locomotor responses in neutrophils in suspension that were dose- and time-dependent. The optimum concentration for the endotoxins of S. dysenteriae, V. cholerae, and K. pneumoniae was 1 mg/ml in which 71, 69, and 66% of the neutrophils were polarized. However, the S. typhimurium dose was 2 mg/ml in which 50% of the cells responded. Neutrophils that were stimulated with endotoxins also showed increased random locomotion (p<0.005) through cellulose nitrate filters, but an enhanced adhesion of the cells to glass surfaces (p<0.03). These are important functions of these cells to reach and phagocytose damaged cells, as well as invading microorganisms. Interestingly, the endotoxins had a highly-significant inhibitory effect upon the proportions of neutrophils phagocytosing opsonized yeast (p<0.01) with a small number of yeast that were engulfed by the cells (p<0.02). Further, endotoxin-treated cells showed an enhanced ability to reduce NBT dye (p<0.03). Therefore, we concluded that endotoxins of enteric pathogens are neutrophil chemotactic factors.

• The Journal of the Korean Society for Microbiology
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• v.35 no.2
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• pp.159-169
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• 2000

Scrub typhus is caused by Orientia tsutsugamushi characterized by fever, headache, lymphadenopathy and eschar formation. Infiltration of inflammatory cells around blood vessels and within the affected organs isS known to be pathologic hallmark of the scrub typhus. Recently, expression of adhesion molecules on vascular endothelial cells was implicated as an important pathogenic mechanism in rickettsial disease. This study was performed to examine the expression of adhesion molecules and to investigate its role in the pathogenesis of O. tsutsugamushi infection. The expression of adhesion molecules on human umbilical vein endothelial cells (HUVEC) was measured by flow cytometry and indirect immunofluorescence. Expression of E-selectin, ICAM-1 and VCAM-1 was significantly increased 4 hours after the infection and persisted at least for 24 hours. Expression of those molecules was not induced by killed O. tsutsugamushi. Adhesion of polymorphonuclear cells and mononuclear cells to HUVEC was increased after the infection with O. tsutsugamushi. In conclusion, adhesion molecules are expressed on HUVEC during the infection of live O. tsutsugamushi and those molecules can contribute to the infiltration of inflammatory cells during the infection.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.10
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• pp.1319-1324
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• 2010

The objectives of this study were to compare the effectiveness of different indicators of mammary inflammation in buffalo and to evaluate the association of the indicators with buffalo milk yield, composition, and rennet coagulation properties. This study was carried out at four buffalo farms in central Italy using a total of 50 lactating buffalo. Milk from each buffalo was tested at the beginning, middle, and end of lactation. To evaluate the relationship between mastitis markers and milk components, three classes were defined for each of the following markers: total somatic cell count (TSCC), differential somatic cell count (DSCC), and bacteriological results The regression coefficient for the reference method and the alternative method of determining TSCC was 0.81, indicating that the method routinely used to analyze buffalo milk consistently underestimated actual TSCC. The milk samples positive for udder-specific bacteria also had higher TSCC values than the samples that were negative for bacteria ($872{\times}10^3$/ml vs. $191{\times}10^3$/ml). In samples that were positive for udder-specific bacteria, polymorphonuclear leukocytes (PMN) made up greater than 50% of the cells. Moreover, only 1% of the samples in the lowest TSCC class were positive for bacteria. The correlation between TSCC and PMN was stronger (0.70), and PMN values in buffalo milk increased significantly when the TSCC class changed from low (38%) to medium and high (56% and 64%). Milk yield was negatively related to TSCC. Significant changes in lactose (4.87%, 4.80% and 4.64%) and chloride content (0.650 mg/ml, 0.862 mg/ml and 0.882 mg/ml) were also observed with increasing TSCC values. Higher TSCC was associated with impaired rennet coagulation properties: the clotting time increased, while the curd firming time ($p{\leq}0.05$) and firmness decreased. We concluded that in buffalo as in dairy cows, TSCC is a valid indicator of udder inflammation; we also confirmed that a value of $ 200{\times}10^3 cells/ml should be used as the threshold value for early identification of an animal affected by subclinical mastitis. In addition to its association with significantly decreased milk yield, a TSCC value above this threshold value was associated with changes in milk composition and coagulating properties.

• 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.