• Parasites, Hosts and Diseases
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• v.32 no.1
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• pp.19-26
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• 1994

An in uipo culturing to examine the cyst stage of ToxopLQsma gondii (ME49 strain) was Investigated using murine peritoneal macrophages, and we also examined the effect of CAMP or DHFR Inhibitors on the growth of bradyzoltes. For experiments ICR mice were Injected 1.p. with 1,500 brain cysts. At 1, 3, 5 and 7 days, peritoneal exudates were Isolated and then adherent peritoneal macrophages were cultured for 1,3,5 and 10 days. Growth pattern of bradyzoltes was measured by (3H)-uracil uptake assay and morphological pattern of pseudocysts formed in macrophages was observed Uth Glemsa stain. Mostly bradyzoites were observed In the macrophages extracted at 3 and S days post Infection. After 3 days in vitro, a number of pseudocysts were formed in the macrophages and the size of pseudocysts was increased during further 5 and 10 days in vitro culture. CAMP stimulated the growth of bradyzoltes when in uiuo 3 and 5 days and then in vitro 5 and 10 days conditions were applied. In case of.DHFR Inhibitors, pynmethamlne produced a linearly decremental effect with a cont.-dependent mode but methotrexate was not effective against Intracellular bradyzoltes or pseudocysts In this system. It was suggested that cyst-forming strain of T gondii (ME49 strain) could be maintained and cultivated in uitro by use of murine peritoneal macrophages. In uivo 3 and 5 days and then in uiko 5 and 10 days conditions appeared to be suitable for culturing of bradyzoltes. CAMP and pyrimethamine had an effect of stimulation and inhibition on the growth of bradyzolte, respectively.

• Nutrition Research and Practice
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• v.11 no.6
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• pp.461-469
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• 2017

BACKGROUND/OBSECTIVE: Airway inflammation by eosinophils, neutrophils and alveolar macrophages is a characteristic feature of asthma that leads to pathological subepithelial thickening and remodeling. Our previous study showed that oxidative stress in airways resulted in eosinophilia and epithelial apoptosis. The current study investigated whether glutathione-containing dry yeast extract (dry-YE) ameliorated eosinophilia, goblet cell hyperplasia and mucus overproduction. MATERIALS/METHOD: This study employed $2{\mu}g$/mL lipopolysaccharide (LPS)- or 20 ng/mL eotaxin-1-exposed human bronchial epithelial cells and ovalbumin (OVA)-challenged mice. Dry-YE employed in this study contained a significant amount of glutathione (140 mg in 100 g dry yeast). RESULTS: Human bronchial epithelial cell eotaxin-1 and mucin 5AC (MUC5AC) were markedly induced by the endotoxin LPS, which was dose-dependently attenuated by nontoxic dry-YE at 10-50 ${\mu}g$/mL. Moreover, dry-YE inhibited the MUC5AC induction enhanced by eotaxin-1, indicating that eotaxin-1-mediated eosinophilia may prompt the MUC5AC induction. Oral supplementation with 10-100 mg/kg dry-YE inhibited inflammatory cell accumulation in airway subepithelial regions with a reduction of lung tissue level of intracellular adhesion molecule-1. In addition, ${\geq}50$ mg/kg dry-YE diminished the lung tissue levels of eotaxin-1, eosinophil major basic protein and MUC5AC in OVA-exposed mice. Alcian blue/periodic acid schiff staining revealed that the dry-YE supplementation inhibited goblet cell hyperplasia and mucus overproduction in the trachea and bronchiolar airways of OVA-challenged mice. CONCLUSIONS: Oxidative stress may be involved in the induction of eotaxin-1 and MUC5AC by endotoxin episode and OVA challenge. Dry-YE effectively ameliorated oxidative stress-responsive epithelial eosinophilia and mucus-secreting goblet cell hyperplasia in cellular and murine models of asthma.

• Journal of Acupuncture Research
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• v.23 no.2
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• pp.73-90
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• 2006

Objectives : The effects of water extract of deer antler herbal-acupunture solution(DHS), prepared from the pilose antler of Cervus korea TEMMINCK var. mantchuricus Swinhoe (Nokyong), a traditional immunosuppressive and immune-activating Korean herbal- acupuncture, on collagen-induced arthritis(CIA:RA model) in mice was studied. Destruction of cartilage and bone are hallmarks of human rheumatoid arthritis, and controlling these erosive processes is the most challenging objective in the treatment of RA. Methods : We investigated the tissue protective effects of deer antler treatment using established murine collagen-induced arthritis(CIA) as a model. Potential synergy of low dosages of anti-inflammatory glucocorticosteroids and deer antler was also evaluated. Results : Treatment of established murine CIA with deer antler herbal-acupunture solution(DHS) $(10-50{\mu}g/day)$ suppressed disease activity and protected against cartilage and bone destruction. Although $10-50{\mu}g/day$ DHS had only a moderate effect on the inflammatory component of the disease activity, it strongly reduced cartilage pathology, as determined by histological examination. Serum cartilage oligomeric matrix protein(COMP) levels were significantly reduced, confirming decreased cartilage involvement. Histological analysis showed that bone destruction was prevented. DHS administration increased serum IL-1Ra levels and reduced anticollagen type II antibody levels. Treatment with low-dose $DHS(1{\mu}g/day)$ was ineffective in suppressing disease score, serum COMP or joint destruction. Synergistic suppression of both arthritis oseverity and COMP levels was noted when low-dose DHS was combined with prednisolone(0.05mg/kg/day), however, which in itself was not effective. Conclusion : DHS was shown to have the inhibiting effects against $IL-1{\alpha}-$ and $IL-1{\beta}-stimulated$ bone resorption. These results indicated that the DAS is not only highly stable and applicable to clinical uses in bone resorption, but also it will be served as a potent anti-inflammatory and anti-arthritic agents for treatment of human RA.

• Toxicological Research
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• v.35 no.1
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• pp.45-63
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• 2019

In view of the growing industrial use of Bacterial cellulose (BC), and taking into account that it might become airborne and be inhaled after industrial processing, assessing its potential pulmonary toxic effects assumes high relevance. In this work, the murine model was used to assess the effects of exposure to respirable BC nanofibrils (nBC), obtained by disintegration of BC produced by Komagataeibacter hansenii. Murine bone marrow-derived macrophages ($BMM{\Phi}$) were treated with different doses of nBC (0.02 and 0.2 mg/mL, respectively 1 and $10{\mu}g$ of fibrils) in absence or presence of 0.2% Carboxymethyl Cellulose (nBCMC). Furthermore, mice were instilled intratracheally with nBC or nBCMC at different concentrations and at different time-points and analyzed up to 6 months after treatments. Microcrystaline $Avicel-plus^{(R)}$ CM 2159, a plant-derived cellulose, was used for comparison. Markers of cellular damage (lactate dehydrogenase release and total protein) and oxidative stress (hydrogen peroxidase, reduced glutathione, lipid peroxidation and glutathione peroxidase activity) as well presence of inflammatory cells were evaluated in brochoalveolar lavage (BAL) fluids. Histological analysis of lungs, heart and liver tissues was also performed. BAL analysis showed that exposure to nBCMC or CMC did not induce major alterations in the assessed markers of cell damage, oxidative stress or inflammatory cell numbers in BAL fluid over time, even following cumulative treatments. $Avicel-plus^{(R)}$ CM 2159 significantly increased LDH release, detected 3 months after 4 weekly administrations. However, histological results revealed a chronic inflammatory response and tissue alterations, being hypertrophy of pulmonary arteries (observed 3 months after nBCMC treatment) of particular concern. These histological alterations remained after 6 months in animals treated with nBC, possibly due to foreign body reaction and the organism's inability to remove the fibers. Overall, despite being a safe and biocompatible biomaterial, BC-derived nanofibrils inhalation may lead to lung pathology and pose significant health risks.

• Reproductive and Developmental Biology
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• v.36 no.1
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• pp.33-37
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• 2012

Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of $Tuj1$ increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous $Igf2$ may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.

• Parasites, Hosts and Diseases
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• v.36 no.1
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• pp.23-32
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• 1998

The pathogenic potential of Acnnthamoebc strains was evaluated by experimental infection of murine AIDS (MAIDS) model. C57BL/6 mice were induced to immunocompromized state by intraperitoneal injection of LP-BM5 MuLV and revealed the typical splenomegalty and Iymphatic enlargement of axillar and inguinal regios on necropsy 4 weeks after viral infection. Although there was no significant difference in the mortality rate of MAIDS mouse according to the culture temperature, it was very different in the mortality rate from strain to strain of Accnthnmoebc. A. henIHi OC-3A strain isolated from the brain of a GAE patient showed !he highest mortality rate and A. culbertsoni A-1 strain from tissue culture was the second. KA/S3 and KA/S2 strains isolated from soil revealed very low virulence. The mice infected by intranasal inoculation of Acanthnmoebc showed relatively chronic course than intravenous inoculation. The gross findings of lungs and brains from infected mice were variable among mice. On the microscopic observations, the lungs showed much more severe inflammation and necrosis than the brains microscopically. This MAIDS model would be useful to study the opportunistic protozoan infections of AIDS patients. In the light of these results. the pathogenic potential and the virulence of Acnnthamoebo may be determined genetically.

• Journal of Periodontal and Implant Science
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• v.45 no.3
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• pp.120-126
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• 2015

Purpose: With the significance of stable adhesion of alveolar bone and peri-implant soft tissue on the surface of titanium for successful dental implantation procedure, the purpose of this study was to apply microgrooves on the titanium surface and investigate their effects on peri-implant cells and tissues. Methods: Three types of commercially pure titanium discs were prepared; machined-surface discs (A), sandblasted, large-grit, acid-etched (SLA)-treated discs (B), SLA and microgroove-formed discs (C). After surface topography of the discs was examined by confocal laser scanning electron microscopy, water contact angle and surface energy were measured. Human gingival fibroblasts (hGFs) and murine osteoblastic cells (MC3T3-E1) were seeded onto the titanium discs for immunofluorescence assay of adhesion proteins. Commercially pure titanium implants with microgrooves on the coronal microthreads design were inserted into the edentulous mandible of beagle dogs. After 2 weeks and 6 weeks of implant insertion, the animal subjects were euthanized to confirm peri-implant tissue healing pattern in histologic specimens. Results: Group C presented the lowest water contact angle ($62.89{\pm}5.66{\theta}$), highest surface energy ($45{\pm}1.2mN/m$), and highest surface roughness ($Ra=22.351{\pm}2.766{\mu}m$). The expression of adhesion molecules of hGFs and MC3T30E1 cells was prominent in group C. Titanium implants with microgrooves on the coronal portion showed firm adhesion to peri-implant soft tissue. Conclusions: Microgrooves on the titanium surface promoted the adhesion of gingival fibroblasts and osteoblastic cells, as well as favorable peri-implant soft tissue sealing.

• IMMUNE NETWORK
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• v.9 no.4
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• pp.138-146
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• 2009

Background: The MHC region of the chromosome contains a lot of genes involved in immune responses. Here we have investigated the mouse NG29/Cd320 gene in the centrometrically extended MHC region of chromosome 17. Methods: We cloned the NG29 gene by RT-PCR and confirmed the tissue distribution of its gene expression by northern blot hybridization. We generated the NG29 gene expression constructs and polyclonal antibody against the NG29 protein to perform the immunofluorescence, immunoprecipitation and flow cytometric analysis. Results: The murine NG29 gene and its human homologue, the CD320/8D6 gene, were similar in the gene structure and tissue expression patterns. We cloned the NG29 gene and confirmed its expression in plasma membrane and intracellular compartments by transfecting its expresssion constructs into HEK 293T cells. The immunoprecipitation studies with rabbit polyclonal antibody raised against the NG29-NusA fusion protein indicated that NG29 protein was a glycoprotein of about 45 kDa size. A flow cytometric analysis also showed the NG29 expression on the surface of Raw 264.7 macrophage cell line. Conclusion: These findings suggested that NG29 gene in mouse extended MHC class II region was the orthologue of human CD320 gene even though human CD320/8D6 gene was located in non-MHC region, chromosome 19p13.

• Archives of Plastic Surgery
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• v.45 no.2
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• pp.111-117
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• 2018

Background Fat grafting, or lipofilling, represent frequent clinically used entities. The fate of these transplants is still not predictable, whereas only few animal models are available for further research. Quantum dots (QDs) are semiconductor nanocrystals which can be conveniently tracked in vivo due to photoluminescence. Methods Fat grafts in cluster form were labeled with cadmium-telluride (CdTe)-QD 770 and transplanted subcutaneously in a murine in vivo model. Photoluminescence levels were serially followed in vivo. Results Tracing of fat grafts was possible for 50 days with CdTe-QD 770. The remaining photoluminescence was $4.9%{\pm}2.5%$ for the QDs marked fat grafts after 30 days and $4.2%{\pm}1.7%$ after 50 days. There was no significant correlation in the relative course of the tracking signal, when vital fat transplants were compared to non-vital graft controls. Conclusions For the first-time fat grafts were tracked in vivo with CdTe-QDs. CdTe-QDs could offer a new option for in vivo tracking of fat grafts for at least 50 days, but do not document vitality of the grafts.

• Journal of Embryo Transfer
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• v.13 no.2
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• pp.179-190
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• 1998

Prolactin (PRL) surge in cycling rats at proestrous afternoon has previously been reported as an inducer of apoptotic cell death of luteal cells. This death-inducing action of PRL seeins unusual, because PRL can he categorized as a cell-survival factor, if other known physiological functions of PRL are taken into account. In this study, the apoptotic action of PRL was assessed in cultured cells prepared from rat luteal tissue and underlying molecular /cellular mechanism of PRL-induced luteolysis was analyzed. The latest crop of corpora lutea (CLs) were enucleated from rat ovaries at 18:00 h on the proestrous day before the next ovulation. Donor rats were pretreated with CB154, a dopamine agonist, in order to he exempted from the endogenous PRL surge. The harvested GLs were dispersed and cultured with or without PRL (2$\mu$g /ml) for 24 or 48 h. An addition of PRL to the culture medium changed the parameters indicative of cell death via apoptosis: a decrease in cell viability (MTT) and an increase in chromatin condensation. Most of the DNA breakdown in nuclei induced by PRL occurred in steroidogenic cells which were identified by 3$\beta$-HSD activity staining, and the number of 3$\beta$-HSD-positivecells were significantly decreased. Interestingly, most of the cells with an apoptotic nucleus adhered to one or more intact and seemingly non-steroidogenic cells. Because the expression of Fas has heen shown to be abundant in murine ovary, and Fas is known to have an exact physiological role in occurrence of apoptotic cell death, the membrane form-Fas ligand (rnFasL) was quantified in the cell lysate. An addition of PRL increased expression of mFasL. Moreover, an addition of concanavalin A (ConA), a T-cell specific activator, in place of PRL, enhanced the apoptotic parameters. Cumulatively, the apoptotic PRL action was addressed to cells unknown than steroidogenic lute~ cells. The most prohable candidate for the direct target cells is Tcells in the luteal tissue that can express mFasL in response to PRL.