• Research in Plant Disease
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• v.22 no.1
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• pp.32-37
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• 2016

In this work, major outer capsid protein (P10) encoded by genome segment S10 of Rice black-streaked dwarf virus (RBSDV) was expressed in Escherichia coli. Genomic dsRNA was extracted from RBSDV-miryang isolate infected rice plants. Based on the sequence of S10 (RBSDV-miryang, GenBank JX994211), a pair of S10 specific primers were designed and used to amplify the fragment encoding the N-part of P10. We amplified the partial gene (S10 1-834 nt) of RBSDV P10 (1-278 aa) by RT-PCR. Amplified RBSDV S10 (1-834 nt) was cloned into the expression vector pET32a (+). Recombinant RBSDV S10 (1-834 nt) was expressed in E. coli BL21(DE3) and purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column. We successfully obtained P10 partial protein of RBSDV and the purified protein was used to immunize rabbits. The resulting polyclonal antiserum specifically recognized RBSDV from infected plant in both Western blotting and enzyme-linked immunosorbent assay. In this study, we provide purified RBSDV P10 (1-278 aa), which would be good material for the serological study of RBSDV-miryang isolates.

• 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 agricultural medicine and community health
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• v.16 no.2
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• pp.165-171
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• 1991

Two patients with confirmed cerebral cysticercosis were treated with Albendazole(Zentel$^{(R)}$) at a daily dose of 1.200mg t.i.d. for 14 consecutive days and evaluated for tolerance and therapeutic effects. First case was 29 year old male, who had experience of 4 times of grand mal seizures during 1 year period before administration in Korea University Hospital. His chief complaints were seizure and moderate degree headache. He also had 4 subcutaneous nodules on the thorax, right and left upper arms. Among them one nodule was biopsied and confirmed microscopically as Cysticercus cellulosae hominis. Computed tomography of the brain showed four round low density lesions in right postero-frontal area, sylvian area, intra-occipital area and left parietal area. Second case was 48 year old male, who also had experience of seizures at 3 years, 5 months and 3 months before administration. In this case, no subcutaneous nodules and no headaches were noted. Brain CT showed four round low density lesions in right postero-parietal area and temporo-parietal area, and left temporo-parietal and parietal area. Serum antibody against cystic fluid antigen was detected by ELISA in both cases. The efficacy of the treatment of cerebral cysticercosis was assessed by the frequency of convulsions after treatment for 22 months follow-up. by the disappearance of the densities in cystic lesions at brain CT for 6 months follow-up, and disappearance of subcutaneous nodules, headache and so on. As the results, all low density lesions in both cases were disappeared in films of brain CT, and 4 nodules in first case were also disappeared. No more seizure and complain of headache occurred during the last 22 months after treatment in both cases. Post-treatment complete blood count and liver function test revealed no remarkable change compared to pre-treatment test. In the nations of Latin America, the physicians do not initially recommended the simultaneous administrations of steroids, reserving them only for patients whom the adverse reactions such as severe headache and/or seizures are occurred. According to them, in most patients these symptoms are controlled with aspirin and symptomatic drugs. But our experience using praziquantel is different, and most cerebral cysticercosis patients who takes PZQ had complaint of severe headache and sometimes seizure. So we simultaneously used dexamethasone as 6mg q.i.d. for 14 consecutive days and 6 days tapering thereafter in both cases for prevention of reactions produced by the host in response to the deaths of the parasites. As the conclusion, albendazole is effective in patients who presented cerebral cysticercosis and albendazole may help in the control of cysticercosis.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.3
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• pp.941-945
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• 2015

Semaphoring is a transmembrane receptor which participates in many cytokine-mediated signal pathways that are closely related to the angiogenesis, occurrence and development of carcinoma. The present study was designed to access the effect of mono-antibody (mAb) guided radioimmunotherapy (RIT) on skin carcinoma and investigate the potential mechanisms. Semaphoring mAb was acquired from mice (Balb/c), purified with rProtein A column; purity, concentration and activity were tested with SDS-PAGE and indirect ELISA; specificity and expression on the cutanuem carcinoma line and tissue were tested by Western blotting; morphology change was assessed by microscopy. MTT assay and colony inhibition tests were carried out to test the influence on the proliferation of tumor cells; Western blotting was also carried out for expression of apoptosis-associated (caspase-3, Bax, Bcl-2) and proliferation-related (PI3K, p-Akt, Akt, p-ERK1/2, ERK1/2) proteins and analyse the change in signal pathways (PI3K/Akt and MEK/ERK). The purity of purified semaphorin mAb was 96.5% and the titer is about $1{\times}10^6$. Western blotting showed semaphoring mAb to have specifically binding stripes with semaphoring b1b2 protein, B16F10, and A431 cells at 39KDa, 100KDa and 130KDa, respectively. Positive expression was detected both in cutanuem carcinoma line and tissue and it mostly located in cell membranes. MMT assay revealed dose-relate and time-relate inhibitory effect of semaphorin mAb on A431 and B16F10. Colony inhibition tests also showed dose-relate inhibitory effects. Western blotting demonstrated the expression of apoptosis and proliferation-related protein and changes in signal pathway. In conclusion, we demonstrated that semaphorin is highly expressed on the tumor cell-surfaces and RIT with semaphorin mAb has effect in i nhibiting proliferation and accelerating apoptosis of tumor cells.

• Parasites, Hosts and Diseases
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• v.30 no.1
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• pp.33-42
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• 1992

Naegleria fowleri, a free-living amoeba commonly found in moist soil and fresh water, enters the body via the nasal mucosa and migrates along the olfactory nerve to t he brain, where it causes acute amoebic meningoencephalitis. In the present study 7 clones secreting monoclonal antibodies (McAbs) against N. fowleri were produced and the effector function of them was investigated. Their isotopes were IgGl (Nf 1, Nf 154), 19G3 (Nf 137) and 19A (Nf 1, Nf 2, Nf 256, Nf 279). Five McAbs (McAb Nf 2, Nf 279, Nf 27, Nf 154, Nf 137) were specific for N. fowleri by ELISA and recognized the antigenic determinants located on the trophoBoite surface by IFAT and immunoperoxidase stain. These aye McAbs had capacity to agglutinate N. fowleri trophozoites and inhibited the growth of the amoeba in culture medium. McAb Nf 2 inhibited proliferation of trophozoites in vitro significantly. Also the cytotoxicity of JV. fowleri against CHO cell was reduced in the presence of McAb Nf 2 and McAb Nf 154. From these results McAb Nf 2 was confirmed to weaken the virulence of the amoeba among 7 screened McAbs.

• Parasites, Hosts and Diseases
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• v.26 no.3
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• pp.169-174
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• 1988

The role of passive cell-mediated transfer of immunity against primary amoebic meningoen- cephalitis(PAME) in mice was studied. Waegleria fowleri, ITMAP 359, were cultured in CGVS medium. The ICR mice used were six week-old males of average weight of 15 g. Immunization was done by three intraperitoneal injections of $1{\times}10^6$ N. fowleri trophozoites at the interval of one week. Splenocytes were obtained from normal and immune mice spleens, and Ix107 cells were administered intraperitoneally into mice 3 days before challenge infection. Mice were infected intranasally with $7{\times}10^4$ N. fowleri trophozoites in a $3{\;}{\mu}l$ suspension under secobarbiturate anesthesia. Transplants of normal or immune splenocytes seem to alter the pattern of the PAME level- opment. The splenocytcs transferred from immune mice reduced the mortality rate in the JV. fowleri infected mice, as compared with the mice transferred with the same number of normal splenocytes or without splenocyte, The blastogenic response of the splenocytes to both lipopoly- saccharide and concanavalin A was elevated on duty 7 after infection the mice transinoculated with immune splenocytes. The serum antibody titers in the mice transferred with immune spleno- cytes were increased gradually from day 7 up to day 20 after infections by mean of ELISA. It is suggested that the transfer of splenocytes from immuniged mice conferred immunity against N. fowleri infection.

• Journal of Plant Biotechnology
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• v.29 no.4
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• pp.287-293
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• 2002

We are trying to develop a transgenic carrot with aims of production and delivery of oral vaccine against microbial enteropathogen using a K88ac pilin gene. A K88ac antigen (pilin) gene was isolated by PCR from the K88ac genomic DNA. The pilin gene was constructed in pGA748 and introduced via Agrobacterium tumefaciens to the explants of carrot hypocotyl and then 494 transgenic lines were established. The amounts of the K88ac antigen produced in each of the cell lines were determined by western and two elite cell lines (M1-17, Y14-1) were selected based on higher levels of expression of the antigens as well as rate of cell growth and efficiency of embryogenesis. In order to test an immunization induced by oral administration of the transgenic carrot, serum of the mice fed with the carrot vaccine were tested in ELISA. It tumed out that the mice fed with 3 g of transgenic carrot showed a similar level of antibody compared to those applied with 10 $\mu\textrm{g}$ of the purified recombinant pilin protein. Besides, various clinical responses were measured after challenging with ETEC K88ac strain to the piglets experiencing an oral immunization with the transgenic carrot. The piglets fed with carrot vaccine showed a lower level of diarrhea in fecal score compared to those fed with non-transgenic carrot. A higher level of increase in weight of the piglets fed with the transgenic carrot vaccine was observed comparing to those fed with non-transgenic carrot as control.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.39
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• pp.8.1-8.8
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• 2017

Background: For an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor $(TGF)-{\beta}1$, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration. Methods: Cytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used. Results: The concentrations of IGF-1, VEGF, and $TGF-{\beta}1$ in MSC-CM were $1515.6{\pm}211.8pg/mL$, $465.8{\pm}108.8pg/mL$, and $339.8{\pm}14.4pg/mL$, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group. Conclusions: VEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.

• The Korea Journal of Herbology
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• v.25 no.2
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• pp.129-136
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• 2010

Objective : Sinapis alba L. (SA) is a korean traditional herbal medicine that is usually used to prevent or treat inflammatory diseases, such as respiratory infection and rheumatoid arthritis. However, the effects of SA supplementation in vitro on serum antibody levels, splenocyte and peritoneal macrophage immune responses have not yet been determined. In this study, we examined the effect of SA on the production of Th1/Th2 cytokines. Methods : Splenocytes were isolated from naive C57BL/6 mice. Cells were enriched for CD4+ cell populations by first staining the cells with anti-CD4 (BD PharMingen, Calif, USA). CD4+ T cells were selected on a (CS) column, and the flow-through was collected as CD4+ T cells. Isolated cells were activated by overnight incubation on 24-well plates coated with $1{\mu}g/mL$ anti-CD3, $1{\mu}g/mL$ anti-CD28 and with SA ($100{\mu}g/mL$). Primary macrophages were collected from the peritoneal cavities of mice (8-week-old female C57BL/6). The peritoneal macrophages were washed and plated with RPMI-1640 overnight for the experiments. After 48-hours cultures, samples were centrifuged at 2000 rpm for 10 minutes, and the supernatants were stored at $-80^{\circ}C$. Mouse IL-4, IFN-$\gamma$ and TNF-$\alpha$ were quantified using ELISA kits (BioSource International, Camarillo, Calif, USA) according to the manufacturer's protocols. Results : SA at 100ug/ml decreased the generation of Th1 cytokine (IFN-$\gamma$) by 0.5-fold. However, SA has no effect on Th2 (IL-4) production. Conclusions : These results suggest that SA may play an important role in the control of T-cell-mediated autoimmunity by down-regulation of Th1 cytokine (especially IFN-$\gamma$, TNF-$\alpha$). These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases.

• Journal of agricultural medicine and community health
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• v.44 no.2
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• pp.82-89
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• 2019

This study was undertaken to determine the seroprevalence and risk factors associated with contracting Lyme disease (LD) among Korean deer farmers. This cross-sectional study devised questionnaire that addressed farm activities, was devised and the blood samples of 516 Korean deer farmers were tested. LD seroprevalence was determined by Western blot test. Fisher's exact test, the chi-squared test, and the chi-squared test for trend analysis were performed to assess the risk associated with LD. Of total 516 study participants recruited, only 12 (2.3%) were seropositive for LD. The result of the study revealed that only deer farmers raising Elk (Cervus Canadensis) were found to be at significantly higher risk of contracting LD than other deer farmers (p=0.033). In addition, a male sex, an age of < 40 years, and those that had raised deer for < 19 years had higher rates of LD infection than their counterparts. Similarly, those that managed livestock sheds, fed deer, processed industry and prepared compost had higher rates of LD infection. In terms of protective factors, those who did not use protective measures such as wear glasses and masks, or protective gloves and aprons, and those that did not disinfect work instruments and did not shower after work had higher rates of LD than those that used protective measures. In conclusion, preventive health strategies should take into account the profiles of deer farmers at greater risk based on considerations of personal, type of work, and the use of personal protective measures.