• Microbiology and Biotechnology Letters
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• v.35 no.2
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• pp.118-127
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• 2007

Germanium-fortified yeast (GY) is a organic germanium-fortified yeast with potent immune modulating activities including anti-inflammatory effect. Through cell line studies, we observed that GY can modulate the diverse immune activity but little evidence was provided on the mechanism of GY in modulating immune activities in other higher animals. In this study, we investigated the effect of GY on modulation of immune function in mice. GY was administered in normal mice or tumor-bearing mice and then effect of GY on modulation of host immune system was analyzed by using ex vivo isolated macrophages, B cells, NK cells. Admistration of GY in mice induced macrophage activation thereby increased effector function of macrophage such as increased phagocytosis, chemotaxis, adherence, $O_2-release$, NO, $TNF-{\alpha}$ production. In addition, GY administration Increased B lymphocyte activation and plaque forming cells. Furthermore, GY administration increased NK-cell mediated cytotoxicity. Furthermore, GY administration suppressed progression of tumor in mice by increasing $TNF-{\alpha}$ production and effector function of NK cells. Our results showed that GY has a potent immunostimulatory function in vivo mice model. Proper modulation and administration of GY in human could be helpful to maintaining immunological homeostasis by modulating host immune system.

• Journal of Microbiology and Biotechnology
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• v.28 no.6
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• pp.997-1006
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• 2018

As shown during the 2009 pandemic H1N1 (A(H1N1)pdm09) outbreak, egg-based influenza vaccine production technology is insufficient to meet global demands during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high-growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). In addition, we confirmed that in the context of the high-growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines that are cheap and can be grown promptly and vigorously as a substitute for egg-based vaccines. Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

• Journal of Microbiology and Biotechnology
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• v.28 no.6
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• pp.849-859
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• 2018

Herpes simplex virus type 2 (HSV-2) infection has been a public health concern worldwide. It is the leading cause of genital herpes and a contributing factor to cervical cancer and human immunodeficiency virus (HIV) infection. No vaccine is available yet for the treatment of HSV-2 infection, and routinely used synthetic nucleoside analogs have led to the emergence of drug resistance. The small molecule $Retro-2^{cycl}$ has been reported to be active against several pathogens by acting on intracellular vesicle transport, which also participates in the HSV-2 lifecycle. Here, we showed that Retro-2.1, which is an optimized, more potent derivative of $Retro-2^{cycl}$, could inhibit HSV-2 infection, with 50% inhibitory concentrations of $5.58{\mu}M$ and $6.35{\mu}M$ in cytopathic effect inhibition and plaque reduction assays, respectively. The cytotoxicity of Retro-2.1 was relatively low, with a 50% cytotoxicity concentration of $116.5{\mu}M$. We also preliminarily identified that Retro-2.1 exerted the antiviral effect against HSV-2 by a dual mechanism of action on virus entry and late stages of infection. Therefore, our study for the first time demonstrated Retro-2.1 as an effective antiviral agent against HSV-2 in vitro with targets distinct from those of nucleoside analogs.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.121-126
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• 1999

Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.295-303
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• 1997

To investigate the response on feeder cell cultures, protoplasts isolated from cell suspensions initiated from mature seed scutellum-derived callus of the Japonica rice variety Taipei 309, were cultured on filter membranes under various conditions. The effects of various factors, such as gelling agents, feeder cell and protoplast densities, species of feeder cells and heat shock treatment, have been investigated to improve protoplast plating efficiencies on filter membranes. Maximum protoplast plating efficiencies were obtained when protoplasts were cultured on KPR medium semi-solidified with Sea Plaque agarose at a density of $5\;\times\;10^{5}\;ml^{-1}$ protoplasts in the presence of Lolium multiflorum as feeder cells (0.5 ml pcv per 10 ml of protoplast culture medium). Pre-culture heat shock treatments for 1 min. and 5 min. to the protoplasts did not give any appreciable increase on the plating efficiency of protoplasts in the presence of feeder cells. Maltose-supplemented medium was superior for plant regeneration from protoplast-derived colonies compared with medium containing only sucrose. The plants were transferred to the glasshouse, flowered and were fertile.

• Journal of Ginseng Research
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• v.42 no.4
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• pp.401-411
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• 2018

Longevity in medicine can be defined as a long life without mental or physical deficits. This can be prevented by Alzheimer's disease (AD). Current conventional AD treatments only alleviate the symptoms without reversing AD progression. Recent studies demonstrated that Panax ginseng extract improves AD symptoms in patients with AD, and the two main components of ginseng might contribute to AD amelioration. Ginsenosides show various AD-related neuroprotective effects. Gintonin is a newly identified ginseng constituent that contains lysophosphatidic acids and attenuates AD-related brain neuropathies. Ginsenosides decrease amyloid ${\beta}$-protein ($A{\beta}$) formation by inhibiting ${\beta}$- and ${\gamma}$-secretase activity or by activating the nonamyloidogenic pathway, inhibit acetylcholinesterase activity and $A{\beta}$-induced neurotoxicity, and decrease $A{\beta}$-induced production of reactive oxygen species and neuro-inflammatory reactions. Oral administration of ginsenosides increases the expression levels of enzymes involved in acetylcholine synthesis in the brain and alleviates $A{\beta}$-induced cholinergic deficits in AD models. Similarly, gintonin inhibits $A{\beta}$-induced neurotoxicity and activates the nonamyloidogenic pathway to reduce $A{\beta}$ formation and to increase acetylcholine and choline acetyltransferase expression in the brain through lysophosphatidic acid receptors. Oral administration of gintonin attenuates brain amyloid plaque deposits, boosting hippocampal cholinergic systems and neurogenesis, thereby ameliorating learning and memory impairments. It also improves cognitive functions in patients with AD. Ginsenosides and gintonin attenuate AD-related neuropathology through multiple routes. This review focuses research demonstrating that ginseng constituents could be a candidate as an adjuvant for AD treatment. However, clinical investigations including efficacy and tolerability analyses may be necessary for the clinical acceptance of ginseng components in combination with conventional AD drugs.

• IMMUNE NETWORK
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• v.6 no.2
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• pp.86-92
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• 2006

Background: Germanium compounds are increased to use in nutrient foods and medicines in terms of antibiotics to microbes, anticancer, modulation of immune system and neutralizing heavy metal toxins. Geranti Bio-Ge Yeast, containing stable organic germanium and bound to the yeast protein was developed by Geranti Pharm. LTD. and the modulation effect in the immune system was examined in vivo and in vitro. Methods: The compound, Geranti Bio-Ge Yeast, was fed to female Balb/c mice (each group has 10 mice) for 4 weeks and the yeast powder and steamed red ginseng powder were used as control during the same feeding time points. During 4 weeks there was no symptom to be considered, and after 4 weeks feeding all mice were sacrificed to check the changes of related immune cells and subsidiary responses (i.e. cell counting, FACS, MTT, LDH, PFC assay). Results: In pre-post comparison, B cell population was increased in the group of Geranti Bio-Ge Yeast in a dose dependent manner (100 to 800 mg/kg). However, the population of T cell, dendritic cell and macrophage was not comparably changed in all doses. The ability of cytokine production and proliferation was almost same level as shown in control group. In contrast, PFC assay informed that the compound increase the antibody production ability when fed over 200 mg/kg implying that the increase of PFC number might be due to the increase of B cells. Conclusion: Over the entire study, we concluded that the compound, Geranti Bio-Ge Yeast has better potential in immune response in terms of B cell proliferation than that of positive control, red ginseng, and the compound can be one of the future candidates for a new supplementary source improving immune system activity.

• The korean journal of orthodontics
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• v.27 no.4 s.63
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• pp.539-548
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• 1997

The purpose of this study was to evaluate the effectiveness of gargling solution with 0.05% NaF and 10% Xylitol in orthodontic patients with fixed appliance. The sample consisted of 20 patients who were classified into an experimental group and a control group, 10 patients each. Experimental group was used experimental gargling solution and the control group was used with placebo solution. The change of S. mutans was analysed by culture on MSB and BHI agar plate pre and post 3, 6, 9 weeks. The results were as follows. 1 There were significant reduction in the number of S. mutans C.F.U. between pre and post 3 weeks(p<0.01), 9 weeks(p<0.05) in experimental group 2. There were significant reduction in the ratio of S. mutans C.F.U. to total C.F.U. between pre and post 3, 6, 9 weeks(p<0.01) in experimental group. 3. S. mutans, which were reduced until 3 weeks, did not show significant change after 3, 6, 9 weeks. 4. S. mutans were strongly suppressed until 3 weeks after gargling solution with 0.05% NaF and 10% Xylitol.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.13-23
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• 2002

The ultimate goal of periodontal therapy is directed to arresting the progression of the disease, and regenerating the fibrous attachment. In order to achieve such treatment aim, the plaque and calculus must be eliminated and the physiological conditions of the root surface must be changed to facilitate the attachment and migration of the new fibroblasts, The method of changing the proper root surface conditions to promote the healing of periodontal tissue involves mechanical procedures, such as scaling and root planing, and chemical procedures such as tetracycline-HCl. However, the formation of a long junctional epithelium was most frequently observed type of healing. Thus, the aim of this study was to examine in vitro the influence of surface conditioning of dentin by TC-HCl on human gingival epithelial cell attachment. Human gingival epithelial cells were obtained from healthy retromolar pad area(under the age 23 years). Seventy two teeth extracted from severe periodontitis were used as study material. To evaluate the epithelial cell attachment to dentin, the prepared specimen was divided to four groups. For the control group, only scaling and root planing were carried out, and for the test group, 1 to 3, the concentration of the TC-HCl was 50, 125 and 250mg/ml respectively. After cell cultivation time of 1-, 3-. 24 hour, for the indirect quantitative assessment of gingival epithelial cell attached to dentin sample, the absorbance of epithelial cell unattached to dentin was measured. The results were as follows; 1. There was no statistically significant difference between scaling and root planing group and TC-HCl 50mg/ml 125mg/ml and 250mg/ml group about absorbance of unattached epithelial cell to dentin sample(p>0.5). 2. As time passes, the absorbance of unattached gingival epithelial cell to dentin sample was decreased statistically significant(p<0.05). 3. There was no statistically significant difference among the TC-HCl group(p>0.05) We concluded that there was similar effect on gingival epithelial cell attachment between TC-HCl conditioning on root surface and only scaling and root planing treatment

• Journal of Periodontal and Implant Science
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• v.30 no.4
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• pp.707-727
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• 2000

매식된 인공치아의 성공을 위해서는 적절한 교합과 수동적 적합성을 갖는 보철물의 제작과 구강내 노출 직후부터의 세균성 치태조절이 요구된다. 본 연구는 전처리(passivation과 tridodecyl - methyl - ammonium chloride(TDMAC) 처리)가 다른 타이타늄 표 면에 chlorhexidine varnish와 테트라사이클린 을 도포시 약제의 방출역학을 알아보고 구강내 치태형성의 억제정도를 평가하기 위하여 시행 되었다. 이를 위해 방출용액으로 인산완충액 성분의 인조타액을 1일${\sim}$1개월간 매일 교환하여 약제농도를 측정하고 타이타늄 박막에 잔류한 약제 활성을 측정하였으며 항균제 도포한 타이타늄 원판을 부착한 장치를 구강내 위치시킨 1일${\sim}$3주 후 원판을 제거하여 주사전자현미경으로 세균 부착상을 관찰하였다. 테트라사이클린은 TDMAC 처리된 표면에서 $10{\sim}18$일까지 유효농도로 방출되었고 표면의 유효 항균 활성은 $3{\sim}4$주간 유지되었으며, chlorhexidine varnish 도포 시에는 TDMAC 전처리시 초기에 $3{\sim}7$일 간 증가한 유효 항균 활성을 방출하여 매식지대치 등에 이러한 항균제도포 시 매식치 주위환경에 항균활성 공급원으로 작용할 수 있음을 보였다. 주사현미경적 관찰시 모든 타이타늄 표면에서 구강내 위치 30분 후에는 세균이 부착되어 있지 않고 타액 단백질 성분에서 유래한 것으로 보이는 피막물질이 표면을 부분 또는 전면에 걸쳐 덮고 있었다. 구강내 노출 2시간 후 항균제 미도포 표본들에는 약간의 구균이 단층으로, $1{\sim}3$일 후에는 부분적으로 두꺼운 세균층을 형성하였고 7일 후에는 표면전체에 걸쳐 세균층이 덮여있었으며 주로 구균과 약간의 간균이 주종을 이루었다. 항균제 도포시 구강내 노출 1주일 이전까지는 미도포군에 비해 치태형성이 지연되는 경향을 보였지만 2주 이후에는 세균 수나 치태형성 양상이 유사하였다. 이 연구로부터 항균제 도포시 1주일 이전의 초기 치태형성을 감소시킬 수 있음을 알 수 있었으며 이러한 연구결과는 타이타늄 임프란트 지대치 표면에 항균제의 도포가 임상적으로 유용할 수 있음을 시사하였다.