• The Korean Journal of Food And Nutrition
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• v.27 no.2
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• pp.267-273
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• 2014

This study found antibacterial activity of $DMfree^{(R)}$ [green tea extract] on facultative bacteria by direct petri dish method and gene array of obligatory M. leprae infected mesenchymal stem cells (MSC). While DMfree showed DPPH radical scavenging effect and high contents of polyphenol, it did not inhibit growth of facultative bacteria such as E. coli and S. aureus on the petri dish. The result does not exclude a possible antibacterial effect of organic solvent extract of green tea rather than DMfree which comes from the water extract of green tea. Pre-treatment of DMfree appeared to have no effect on copy number of 14 genes compared with control MSC by real-time RT-PCR. However pre-treatment of DMfree on M. leprae infected MSC revealed a significant decrease of anti-inflammatory cytokine (IL-6), (P<0.038) and sharp down-regulation of pro-inflammatory cytokine (IL-1). Enhanced expression of VEGFR-1 mRNA was noted in DMfree pretreated M. leprae infected MSC group (P<0.003). These results show that DMfree would stabilize M. leprae infected MSC from further inflammation by down-regulating anti-inflammatory cytokine (IL-6) and pro-inflammatory cytokine (IL-$1{\beta}$). This is the first report on DMfree inhibition of IL-6 and IL-$1{\beta}$ expression in M. leprae infected MSC. Further experiments that detect protein levels of IL-$1{\beta}$ and IL-6 may support the result of this gene array.

• Parasites, Hosts and Diseases
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• v.56 no.3
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• pp.229-236
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• 2018

Cutaneous leishmaniasis (CL) has been one of the most common parasitic diseases in Saudi Arabia. This study exhibits the clinical features, diagnosis, cytokine profile and treatment of CL patients in Al-Taif province. Ninety CL suspects at a tertiary care general hospital were enrolled in one-year study. Patients were interviewed, clinically-examined, and subjected to laboratory tests: skin scraping smear microscopy, OligoC-TesT commercial PCR (Coris BioConcept) and kinetoplast DNA (kDNA) PCR for Leishmania diagnosis. Interferon-gamma (RayBio; Human $IFN-{\gamma}$ ) and nitric oxide (NO) levels in patients' sera were evaluated before treatment with sodium stibogluconate (pentostam) with 20-day intramuscular drug regimen. Positive rates of microscopy, commercial PCR and kDNA PCR were 74.4%, 95.5% and 100%, respectively. Patients came to hospital mostly in winter (45.0%). CL was frequently exhibited in Saudi patients (78.8%), male gender (70.7%), age <20 years (50.0%), rural-dwellers (75.5%) and patients with travel history (86.6%). Lesion was mostly single ulcer (93.3%), occurred in the face (67.7%). Upon pentostam treatment, 85.1% of ulcers showed rapid healing signs. Levels of $IFN-{\gamma}$ and NO were significantly higher in the healing than the non-healing cases (P<0.001). The kDNA PCR proved more sensitive than microscopy and OligoC-TesT commercial PCR. Our results open perspectives for $IFN-{\gamma}$ use as a biomarker predicting treatment response.

• Archives of Pharmacal Research
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• v.29 no.10
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• pp.890-897
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• 2006

In inflammatory responses, induction of cytokines and other immune regulator genes in macrophages by pathogen-associated signal such as lipopolysaccharide (LPS) plays a crucial role. In this study, the gene expression profile changes by LPS treatment in the macrophage/monocyte lineage cell line RAW264.7 was investigated. A 60-mer oligonucleotide microarray of which probes target 32381 mouse genes was used. A reverse transcription-in vitro translation labeling protocol and a chemileuminescence detection system were employed. The mRNA expression levels in RAW264.7 cells treated for 6 h with LPS and the control vehicle were compared. 747 genes were up-regulated and 523 genes were down-regulated by more than 2 folds. 320 genes showing more than 4-fold change by LPS treatment were further classified for the biological process, molecular function, and signaling pathway. The biological process categories that showed high number of increased genes include the immunity and defense, the nucleic acid metabolism, the protein metabolism and modification, and the signal transduction process. The chemokine-cytokine signaling, interleukin signaling, Toll receptor signaling, and apoptosis signaling pathways involved high number of genes differentially expressed in response to LPS. These expression profile data provide more comprehensive information on LPS-target genes in RAW264.7 cells, which will be useful in comparing gene expression changes induced by extracts and compounds from anti-inflammatory medicinal herbs.

• International Journal of Stem Cells
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• v.16 no.2
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• pp.244-249
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• 2023

Background and Objectives: To examine whether ischemic retinal ganglion cells (RGCs) will be salvaged from cell death by human adipose-derived mesenchymal stem cells (ADSCs) in an organotypic retina model. Methods and Results: Deprived of arterial oxygen supply, whole mice retinas were cultured as an ex vivo organotypic cultures on an insert membrane in a 24-well plate. The therapeutic potential of ADSCs was examined by co-culture with organotypic retinas. ADSCs were seeded on top of the RGCs allowing direct contact, or at the bottom of the well, sharing the same culture media and allowing a paracrine activity. The number of surviving RGCs was assessed using Brn3a staining and confocal microscopy. Cytokine secretion of ADSCs to medium was analyzed by cytokine array. When co-cultured with ADSCs, the number of surviving RGCs was similarly significantly higher in both treatment groups compared to controls. Analysis of ADSCs cytokines secretion profile, showed secretion of anti-apoptotic and pro-proliferative cytokines (threshold>1.4). Transplantation of ADSCs in a co-culture system with organotypic ischemic retinas resulted in RGCs recovery. Since there was no advantage to direct contact of ADSCs with RGCs, the beneficial effect seen may be related to paracrine activity of ADSCs. Conclusions: These data correlated with secretion profile of ADSCs' anti-apoptotic and pro-proliferative cytokines.

• IMMUNE NETWORK
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• v.7 no.2
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• pp.66-74
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• 2007

Background: The genus Flavivirus consists of many emerging arboviruses, including Dengue virus (DV), Japanese encephalitis virus (JEV) and West Nile virus (WNV). Effective preventive vaccines remain elusive for these diseases. Mice are being increasingly used as the animal model for vaccine studies. However, the pathogenic mechanisms of these viruses are not clearly understood. Here, we investigated the interaction of DV and JEV with murine bone marrow-derived dendritic cells (bmDC). Methods: ELISA and FACS analysis were employed to investigate cytokine production and phenotypic changes of DCs obtained from bone marrow following flavivirus infection. Results: We observed that these viruses altered the cytokine profile and phenotypic markers. Although both viruses belong to the same family, JEV-infected bmDC produced anti-inflammatory cytokine (IL-10) along with pro-inflammatory cytokines, whereas DV infection induced production of large amounts of pro-inflammatory cytokines (IL-6 and TNF-${\alpha}$) and no IL-10 from murine bmDCs. Both flaviviruses also up-regulated the expression of co-stimulatory molecules such as CD40, CD80 and CD86. JEV infection led to down-regulation of MHC II expression on infected bmDCs. We also found that cytokine production induced by JEV and DV is MyD88-dependent. This dependence was complete for DV, as cytokine production was completely abolished in the absence of MyD88. With regard to JEV, the absence of MyD88 led to a partial reduction in cytokine levels. Conclusion: Here, we demonstrate that MyD88 plays an important role in the pathogenesis of flaviviruses. Our study provides insight into the pathogenesis of JEV and DV in the murine model.

• Biomedical Science Letters
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• v.19 no.2
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• pp.142-146
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• 2013

Enterotoxigenic Bacteroides fragilis (ETBF) causes inflammatory diarrhea in humans and animals and is also implicated in colorectal cancer. ETBF-infected mice exhibit a prominent large intestinal inflammation characterized by neutrophil infiltration and induction of the Th17 response. In this study, we examined differences in the secreted cytokine profile of the cecum and proximal colon of ETBF-infected mice using an antibody array. Of the cytokines examined, we found that the cecal tissues from ETBF-infected mice secreted elevated levels of G-CSF, IL-6, IL-17 and LIX compared to non-toxigenic Bacteroides fragilis (NTBF) and Mock infected mice. The proximal colon tissues from ETBF-infected mice secreted higher levels of G-CSF, IL-6, KC, LIX, MIP-1g and MCP-1. This study demonstrates that the cecum and colon should be considered separately when assays are used to determine immune responsiveness to enteric infections.

• YAKHAK HOEJI
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• v.59 no.4
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• pp.170-176
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• 2015

In vaccine development, the major points may be induction of effective and increased levels of antibody production. This is especially the case when the antigenic sources are carbohydrates. For many years, thus, we have researched various types of formulations such as liposomal and conjugate vaccines. However, the fastidious formulation process and high costs are a problem. For this reason, there is currently a focus on utilizing immunoadjuvants. In this present study, we tested if platycodin D (PLD) from Platycodon Radix have immunoadjuvant activity against the cell wall of Candida albicans (CACW). The resulting data showed that in the murine model of antibody production, CACW combined with PLD [CACW/PLD/IFA] increased the production of antibodies specific to C. albicans when compared to the antibody production by [CACW/IFA]-induction, which was used as a negative control (P<0.05). In the case of [CACW/PLD/IFA], the antibody production was 1.4 times as that of the CFA. In addition, formulations containing either had a prolonged antibody inducing activity maintaining the initial titers of antibody as compared to the CFA formula. Cytokine profiling with the antisera displayed that the PLD produced both Th1 and Th2 immunoresponses, but Th1 dominant was much greater (P<0.05). Furthermore, [CACW/PLD/IFA] formula enhanced resistance of mice against disseminated candidiasis, whereas the CFA had no such effect. In conclusion, PLD has an immunologic activity, which is protective against the disease. Thus, PLD can be a goof candidate for a new immunoadjuvant in development of the fungal vaccine.

• Nutrition Research and Practice
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• v.12 no.6
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• pp.479-485
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• 2018

BACKGROUND/OBJECTIVES: Gelidium amansii (GA) contains plenty of agars and various biological substances, which make them a popular functional food to control body weight in previous studies. Unlike previous studies focused on agar in GA, objectives of this study were to investigate the effects of agar-free GA extract (AfGAE) on preventive and treatment models by using diets-induced obese (DIO) C57BL/6J mice. MATERIALS/METHODS: AfGAE were used to test their effects on the prevention (Exp-1) and treatment (Exp-2) against obesity after pilot study in DIO mice. The weight changes of the body and fat tissues and protein expression related to lipid metabolism and inflammation as well as plasma lipid profile and insulin were detected. RESULTS: Although AfGAE did not prevent long-term DIO, it did increase the levels of anti-inflammatory cytokine production and lipolysis protein. We further evaluated various doses of AfGAE in preventive and treatment models. As a result, our findings suggested that an AfGAE administration as a preventive model might be a better approach to achieve its anti-inflammatory and lipolysis-promoting effects in DIO mice. CONCLUSION: Although future studies to investigate the target materials such as polyphenols in AfGAE are required, the result suggests that GA without agar might be a therapeutic tool to improve health conditions related to inflammation.

• The Korea Journal of Herbology
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• v.27 no.3
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• pp.75-82
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• 2012

Objective : In the present study, we investigated the pharmacological profile of the aqueous extract of $Patrinia$ $scabiosaefolia$ Fischer ex Link (EPS) and $Patrinia$ $villosa$ Jussieu (EPV) for its anti-asthmatic activities. The purpose of this study is to ascertain if EPS result in better anti-asthmatic activities and functional outcome as compared with EPV. Methods : In this study, BALB/c mice were systemically sensitized to ovalbumin (OVA) followed intratracheally, intraperitoneally, and by aerosol allergen challenges. We investigated the effect of EPS, EPV on the recruitment of pulmonary inflammatory cells, various immune cell phenotypes, Th1/Th2 cytokine gene expression and production and histamine production in serum. Results : In BALB/c mice, we found that EPV-treated groups had more effectively suppressed inflammatory cell infiltration of lung and BALF, B220+IgE+, CD11b+Gr-1+ cell population in lung and these occurred by suppressing the gene expression of IL-4, IL-5 and IL-4 cytokine production in BALF and serum. Conclusions : These results suggest that EPV may play an important role in the control of anti-asthmatic activities by down-regulation of Th2 cytokine (especially IL-4, IL-5). In general, EPV has shown a better anti-asthmatic activities compared to EPS.

• Biomolecules & Therapeutics
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• v.20 no.2
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• pp.171-176
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• 2012

HaCaT cells are the immortalized human keratinocytes and have been extensively used to study the epidermal homeostasis and its pathophysiology. T helper cells play a role in various chronic dermatological conditions and they can affect skin barrier homeostasis. To evaluate whether HaCaT cells can be used as a model cell system to study abnormal skin barrier development in various dermatologic diseases, we analyzed the gene expression profile of epidermal differentiation markers of HaCaT cells in response to major T helper (Th) cell cytokines, such as $IFN{\gamma}$, IL-4, IL-17A and IL-22. The gene transcriptional profile of cornified envelope-associated proteins, such as filaggrin, loricrin, involucrin and keratin 10 (KRT10), in HaCaT cells was generally different from that in normal human keratinocytes (NHKs). This suggests that HaCaT cells have a limitation as a model system to study the pathophysiological mechanism associated with the Th cell cytokine-dependent changes in cornified envelope-associated proteins which are essential for normal skin barrier development. In contrast, the gene transcription profile change of human ${\beta}2$-defensin (HBD2) in response to $IFN{\gamma}$, IL-4 or IL-17A in HaCaT cells was consistent with the expression pattern of NHKs. $IFN{\gamma}$ also up-regulated transglutaminase 2 (TGM2) gene transcription in both HaCaT cells and NHKs. As an alternative cell culture system for NHKs, HaCaT cells can be used to study molecular mechanisms associated with abnormal HBD2 and TGM2 expression in response to $IFN{\gamma}$, IL-4 or IL-17A.