• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.769-773
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• 2007

With the aim to produce ascorbic acid-2-phosphate(AsA-2-P) from L-ascorbic acid(AsA, Vitamin C), nine bacteria conferring the ability to transform AsA to AsA-2-P were isolated from soil samples alongside known strains from culture collections. Most isolates were classified to the genus Brevundimonas by 16S phylogenetic analysis. Among them, Brevundimonas diminuta KACC 10306 was selected as the experimental strain because of its the highest productivity of AsA-2-P. The optimum set of conditions for the AsA-2-P production from AsA using resting cells as the source of the enzyme was also investigated. The optimum cultivation time was 16 h and the cell concentration was 120g/l(wet weight). The optimum concentrations of AsA and pyrophosphate were 550mM and 450mM, respectively. The most effective buffer was 50mM sodium formate. The optimum pH was 4.5 and temperature was $40^{\circ}C$. Under the above conditions, 27.5g/l of AsA-2-P was produced from AsA after 36 h of incubation, which corresponded to a 19.7% conversion efficiency based on the initial concentration of AsA.

• Biomolecules & Therapeutics
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• v.19 no.4
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• pp.425-430
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• 2011

In this study, we found that Forsythiae fructus (FF) and one of its main compounds, arctigenin, significantly inhibited nitric oxide production in lipopolysaccharide (LPS)-stimulated BV-2 microglial cells. Arctigenin also suppressed the expression of inducible nitric oxide synthase and cyclooxygenase-2, and inhibited the activation of extracellular signal-regulated kinase, c-Jun N-terminal kinase and p38. Moreover, it also reduced levels of proinflammatory cytokines, interleukin $1{\beta}$, tumor necrosis factor ${\alpha}$ and prostaglandin E2, and inhibited neuronal death in LPS-treated organotypic hippocampal cultures. Therefore, we suggest that arctigenin may confer a neuroprotective effect via the inhibition of neuroinflammation.

• The Korea Journal of Herbology
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• v.25 no.3
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• pp.19-25
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• 2010

Objective : The aim of this study was to investigate anti-inflammatory activity of SD-01 methanol extract in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods : Cytotoxic activity of SD-01 methanol extract on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines and $PGE_2$ were measured by ELISA method. The levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), $I{\kappa}$-B-alpha and nuclear NF-${\kappa}$ B p65 expression were detected by western blot. Results : Our results indicated that methanol extract of SD-01 significantly inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-$\alpha$, IL-$1\beta$, IL-6 and MCP-1 production in RAW 264.7 cells. Moreover, methanol extract of SD-01 treatment also blocked LPS-induced NF-kB activation. Conclusion : These findings indicate that methanol extract of SD-01 inhibits the production of pro-inflammatory mediators and cytokines via suppression of NF-${\kappa}$ B activation. Take together, these results indicate that methanol extract of SD-01 has the potential for use as an agent of anti-chronic inflammatory diseases.

• The Journal of Internal Korean Medicine
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• v.19 no.2
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• pp.261-277
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• 1998

The purpose of this research was to investigate cytotoxicity of DangkwiYonghoe-Hwan(DYH) and the constitutive crude drugs on several cancer cell-lines, thymocytes, splenocytes and 3T3 cells. The DYH consists of Coptidis Rhizoma, Scutellariae Radix, Phellodendri Cortex, Gardeniae Fructus, Gentianae scabrae Radix, Indigo pulverata Levis, Aloe, Rhei Rhizoma, Moschus, Saussureae Radix and Angelicae Gigantis Radix. The cytotoxicity was determined by MTT method. The DYH inhibited the proliferation of MOLT-4, K562, HL-60, Jurkat, L1210, P815, S180 and Yac-1, thymocyte, splenocyte and 3T3 cells. The cytotoxicity of Coptidis Rhizoma on the cancer cell-lines was the most potent in the constitutive crude drugs. The proliferation of cancer cell-lines was partly inhibition and partly increase by the treatment of Scutellariae Radix, Gardeniae Fructus, Gentianae scabrae Radix, Indigo pulverata Levis, Aloe, Rhei Rhizoma, Moschus and Angelicae Gigantis Radix. Phellodendri Cortex and Saussureae Radix had a poor cytotoxicity on cancer cell-lines. Coptidis Rhizoma and Phellodendri Cortex inhibited the proliferation of thymocyte, splenocyte and 3T3 cells.

• Nuclear Medicine and Molecular Imaging
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• v.42 no.5
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• pp.394-400
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• 2008

Purpose: Quantitative comparison of transgene expression within stem cells between lentivirus and adenovirus-mediated delivery systems has not been reported. Here, we evaluated the human sodium iodide symporter (hNIS) gene expression in rat mesenchymal stem cell (rMSC) transduced by lentivirus or adenovirus, and compared the hNIS expression quantitatively between the two delivery systems. Materials and Methods: Lentiviral-mediated hNIS expressing rMSC (lenti-hNIS-rMSC) was constructed by cloning hNIS gene into pLenti6/UbC/V5-DEST (Invitrogen) to obtain pLenti-hNIS, transducing rMSC with the pLenti-hNIS, and selecting with blasticidin for 3 weeks. Recombinant adenovirus expressing hNIS gene (Rad-hNIS) was produced by homologous recombination and transduction efficiency of Rad-hNIS into rMSC evaluated by Rad-GFP was $19.1{\pm}4.7%$, $54.0{\pm}6.4%$, $85.7{\pm}8.7%$, and $98.4{\pm}1.3%$ at MOI 1, 5, 20, and 100, respectively. The hNIS expressions in lenti-hNIS-rMSC or adeno-hNIS-rMSC were assessed by immunocytochemistry, western blot, and 1-125 uptake. Results: Immunocytochemistry and western blot analyses revealed that hNIS expressions in lenti-hNIS-rMSC were greater than those in adeno-hNIS-rMSC at MOI 20 but lower than at MOI 50. However in vitro 1-125 uptake test demonstrated that iodide uptake in lenti-hNIS-rMSC ($29,704{\pm}6,659\; picomole/10^6\;cells$) was greater than that in adeno-hNIS-rMSC at MOI 100 ($6,168{\pm}2,134\;picomole/10^6\;cells$). Conclusion: Despite lower amount of expressed protein, hNIS function in rMSC was greater by lentivirus than by adenovirus mediated expression. Stem cell tracking using hNIS as a reporter gene should be conducted in consideration of relative vector efficiency for transgene expression.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.3
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• pp.232-238
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• 2020

This study used adult wistar-based rats to observe the sexual cycle as a morphological characteristic of vaginal epithelial cells by vaginal smearing, and investigated the fetal number through mating with male rats of the same strain. The target animal was a 12 to 13-week-old Wistar-based mature unlighted rat (weight 220 g to 240 g), room temperature 23 ± 2℃, 14 hours artificial lighting (05:00 to 19:00 hours), 10 hours Adapted individuals were used for rearing for at least 2 weeks under the conditions of the darkroom (19:00 to 05:00). The feed was managed for free feeding of pellet feed for animals and water. The vaginal smearing method was used for the experiments by observing the sexual cycle every morning and confirming that the normal sexual cycle of 4 or 5 days was repeated at least 2 cycles or more. As a result, the proestrus was found to have few red blood cells, the cells and nuclei were rather large and round, and many nucleated cells were identified. In the case of the estrus, the cells were large and the nuclei were not stained, and most of the keratinocytes were found. In addition, in the metestrus and diestrus, there were many white blood cells, and it was confirmed that nucleated epithelial cells and keratinocytes were significantly reduced. The pregnancy period was 21 ± 1.8 days, and the number of live births per delivery was 11.9 on average. The number of fetuses on the 8th and 10th days of pregnancy were 15.2 ± 0.4 and 15.4 ± 0.3, respectively. On the contrary, the number of fetuses on the 12th day of pregnancy was 12.9 ± 0.6, which was significantly (p < 0.05) decreased compared to the 10th day of pregnancy, and the number of fetuses was similar until delivery. As a result of investigating the change of body weight according to the birth weight and growth stage after delivery, the birth weight of female and male was 9.2 ± 2.0 g and 9.8 ± 2.5 g, respectively. After that, until the 16th day, the female and the male showed similarly moderate weight gain, and then showed a rapid weight gain until the 21st day of lactation. With reference to the results of this study, it is expected to be used as basic data for determining the mating time of rodents and controlling pregnancy and fetal number.

• Journal of Gastric Cancer
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• v.5 no.2
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• pp.106-112
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• 2005

Purpose: Considering that nutritional state correlates to immunity, we performed this study to evaluate the correlation by assessing the numerical changes of the levels of serum albumin and lymphocyte subsets. Materials and Methods: The study was performed on patients who were diagnosed as having gastric cancer and who underwent curative surgery from August 1998 to August 2004 in the Gyeongsang National University Hospital and whose peripheral blood lymphocyte subsets were tested prior to surgery. The study population was a total of 150 cases. Results: The change in the lymphocyte subsets in relation to the change in the level of serum albumin in all patients with gastric cancer was determined, and was compared to disease stages. When patients were classified by using the level of serum albumin with 3.2 mg/dl as the cut-off point (low group: serum albumin <3.2 mg/dl, normal group = serum albumin $\geq$ 3.2 mg/dl), the number of peripheral blood lymphocytes, CD3+ cells, CD4+ cells, CD8+ cells, and CD16+ 56 cells were, significantly lower in the group with the level of serum albumin below 3.2 mg/dl (low group) than it was in the group with a serum albumin level above 3.2 mg/dl (normal group) (P<0.05). In stage I (n=59), CD16+56 cells were significantly lower in the low group. In stage II (n=29), the number of CD16+56 cells was lower and the ratio of CD4+/CD8+ was higher in the low group than in the normal group significantly. In stage IV (n=33), except for CD19+ cells, the number of all lymphocyte subsets was significantly lower and the ratio of CD4+/CD8+ was significantly higher in the low group. Conclusion: The group with a low level of serum albumin had a low absolute number of lymphocyte subsets. Based on this, we reconfirmed that the nutritional state is closely related with the immune state in patients with gastric cancer.

• Biomolecules & Therapeutics
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• v.19 no.3
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• pp.288-295
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• 2011

Amyloid beta ($A{\beta}$)-induced neurotoxicity is a major pathological mechanism of Alzheimer's disease (AD). In this study, we investigated the inhibitory effect of L-theanine, a component of green tea (Camellia sinensis) on $A{\beta}_{1-42}$-induced neurotoxicity and oxidative damages of macromolecules. L-theanine inhibited $A{\beta}_{1-42}$-induced generation of reactive oxygen species, and activation of extracellular signal-regulated kinase and p38 mitogenic activated protein kinase as well as the activity of nuclear factor kappa-B. L-theanine also signifi cantly reduced oxidative protein and lipid damage, and elevated glutathione level. Consistent with the reduced neurotoxic signals, L-theanine (10-50 ${\mu}g$/ml) concomitantly attenuated $A{\beta}_{1-42}$ (5 ${\mu}M$)-induced neurotoxicity in SK-N-MC and SK-N-SH human neuroblastoma cells. These data indicate that L-theanine on $A{\beta}$-induced neurotoxicity prevented oxidative damages of neuronal cells, and may be useful in the prevention and treatment of neurodegenerative disease like AD.

• Molecules and Cells
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• v.44 no.1
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• pp.13-25
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• 2021

Apoptosis and compensatory proliferation, two intertwined cellular processes essential for both development and adult homeostasis, are often initiated by the mis-regulation of centrosomal proteins, damaged DNA, and defects in mitosis. Fly Anastral spindle 3 (Ana3) is a member of the pericentriolar matrix proteins and known as a key component of centriolar cohesion and basal body formation. We report here that ana3m19 is a suppressor of lethality induced by the overexpression of Sol narae (Sona), a metalloprotease in a disintegrin and metalloprotease with thrombospondin motif (ADAMTS) family. ana3m19 has a nonsense mutation that truncates the highly conserved carboxyl terminal region containing multiple Armadillo repeats. Lethality induced by Sona overexpression was completely rescued by knockdown of Ana3, and the small and malformed wing and hinge phenotype induced by the knockdown of Ana3 was also normalized by Sona overexpression, establishing a mutually positive genetic interaction between ana3 and sona. p35 inhibited apoptosis and rescued the small wing and hinge phenotype induced by knockdown of ana3. Furthermore, overexpression of Ana3 increased the survival rate of irradiated flies and reduced the number of dying cells, demonstrating that Ana3 actively promotes cell survival. Knockdown of Ana3 decreased the levels of both intra- and extracellular Sona in wing discs, while overexpression of Ana3 in S2 cells dramatically increased the levels of both cytoplasmic and exosomal Sona due to the stabilization of Sona in the lysosomal degradation pathway. We propose that one of the main functions of Ana3 is to stabilize Sona for cell survival and proliferation.

• The Korea Journal of Herbology
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• v.33 no.3
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• pp.19-24
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• 2018

Objectives : Hedgehog skin is one of the animal medicines in Traditional Korean Medicine for hematochezia and hemorrhoids. In this study, we examined cytotoxicity and anti-inflammatory effects. Methods : Cytotoxicity of hedgehog skin extracts was measured by MTT assay in vitro. We investigated the inhibition of lipopolysaccharide (LPS) stimulated nitric oxide (NO) production in RAW 264.7 cells. The phosphorylation of mitogen-activated protein kinases (MAPKs) was measured by western blot. And we observed the effect of hedgehog skin extracts on the expression of IL-6 genes using real time PCR. Results : As a result of MTT assay for cytotoxicity, there were no significant differences between non-treatment group and hedgehog skin extracts treatment groups. $500{\mu}g/m{\ell}$ of hedgehog skin extracts treatment significantly decreased nitric oxide production in comparison with non-treatment in LPS-induced RAW 264.7 cells. In measurement of the phosphorylation of MAPKs using western blot analysis, LPS stimulation increased the phosphorylation of MAPKs and $500{\mu}g/m{\ell}$ of hedgehog skin extracts treatment decreased the phosphorylation of ERK1, ERK2 and p38 significantly. But there were no significant differences the phosphorylation of JNK1 and JNK2. As a result of confirmation of the IL-6 mRNA gene expression using real time PCR, IL-6 mRNA gene expressions were significantly decreased in $50{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$ and $500{\mu}g/m{\ell}$ hedgehog skin extracts treated groups by comparison with non-treatment group. Conclusion : These results could provide a mechanistic explanation for the anti-inflammatory effects of the hedgehog skin.