• Clinical and Experimental Pediatrics
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• v.45 no.2
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• pp.247-255
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• 2002

Purpose : This study was undertaken to obtain basic data about the megakaryocyte colony formation of fetal liver cells by using immunocytochemical staining and ex vivo culture with growth factors. Methods : The mononuclear cells were isolated from fetal liver and bone marrow with idiopathic thrombocytopenic purpura(ITP) and pancytopenia. These mononuclear cells were cultured in $MegaCult^{TM}-C$(Stem Cell Tech, Canada) media in the presence of growth factors and CFU-Megakaryocyte( CFU-Mk) colonies were counted on day 12. The expansion of CD34+ and CD41+ cell was analyzed by flow cytometry after 5 days incubation using flask culture. Results : The numbers of CFU-Mk colonies of mononuclear cells obtained from fetal liver in the 11th week gestational age were more than those in the 19th week specimens; growth factors could not enhance the colony expansion in all cases. Total numbers of CFU-Mk colony of fetal liver cells were higher than bone marrow from ITP or pancytopenia groups. The numbers of pure or large CFU-Mk colonies of fetal liver cells were also higher than bone marrow specimens. The rate of CD34+ cell expression of fetal liver was increased after flask culture and the enhancement effect of epression was seen only in cases which added thrombopoietin. The rate of CD41+ cell expression of fetal liver was increased after incubation, but the enhancement effect of growth factors was unclear. Conclusion : This study revealed good results about the megakaryocyte colony assay of fetal liver mononuclear cells using $MegaCult^{TM}-C$ media. This study suggests that the fetal liver could be a good source of megakaryocytic progenitor cells for clinical application in hematopoietic stem cell transplantation.

• Korean Journal of Veterinary Service
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• v.20 no.1
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• pp.11-18
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• 1997

Characteristics, pathogenicity and control of the causative organisms isolated front diseased cultured flounder, Paralichthys olivaceus were studied. The causative organisms were identified as E tarda by biochemical and biophysical characteristics. Also, it strains were named as E tarda KBF-1 and E tarda KMF-1, and optimal pH of E tarda KBF-1 and E tarda KMF-1 were 8.0, and optimal concentration of NaCl. E tarda KBF-1 was 0% and E tarda KMF-1 was 1%. In the pathogenicity test, 0~10 of the flounders of artificially infected group(E tarda KBF-1) with $1.0{\times}10^7$ cfu/fish were died within 60 hrs, but 0~9 flounders infected group with 41.0{\times}10^6$ cfu/fish were died within 60 hrs. Also, 0~10 flounders infected group(E tarda KMF-1) with $1.0{\times}10^7$ cfu/fish were died within 36 hrs, while 0~7 flounders infected with $1.0{\times}10^6$ cfu/fish were died within 60 hrs. Drug sensitivity of E tarda KBF-1 strain was resistant to AM, CF and N, and intermediate to E, K and S, and sensitivity to C, G, SxT and FF. But E tarda KMF-1 strain was resistant to CF, E and V, and intermediate to AM, C, N and SxT, and sensitivity to GM and FF.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.24 no.3
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• pp.300-309
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• 2014

Objectives: The objective of this study is to evaluate microbial exposure hazards in the metal-working fluids(MWF) handling industry. Methods: Air quality parameters(airborne bacteria, fungi, endotoxin and oil mist) and bulk MWF in storage tanks were evaluated at 54 points at nine sites in South Korea. Results: The geometric means(GM) of culturable airborne bacteria, fungi, endotoxin and oil mist concentration were $133CFU/m^3$(n=376, range $7{\sim}6,510CFU/m^3$), $159CFU/m^3$(n=381, range $7{\sim}8,469CFU/m^3$), $8.06EU/m^3$(n=103, range $0.34{\sim}280.4EU/m^3$) and $0.20mg/m^3$(n=104, range $0.01{\sim}2.87mg/m^3$), respectively. The ratio of indoor to outdoor concentration was 2.7 for bacteria, 6.1 for endotoxin, and 4.8 for oil mist. Even though average airborne bacteria concentration did not exceed recommended exposure limits($1,000CFU/m^3$), MWF in the storage tanks was highly contaminated with bacteria(arithmetic mean $2.1{\times}10^6CFU/ml$) and exceeded recommended bacteria limits($10^5CFU/ml$). Conclusions: It is necessary for MWF handling workplaces to conduct periodical biohazard inspection of MWF storage tanks. Additionally, further research may be necessary to establish biological occupational exposure limits.

• Journal of Environmental Health Sciences
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• v.37 no.6
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• pp.406-417
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• 2011

Objectives: Exposure to bioaerosols in the indoor environment could be associated with a variety adverse health effects, including allergic disease such atopy. The objectives of this study were to assess children's exposure to bioaerosol in home indoor environments and to evaluate the association between atopy and bioaerosol, environmental, and social factors in Ulsan, Korea. Methods: Samples of viable airborne bacteria and fungi were collected by impaction onto agar plates using a Quick Take TM 30 and were counted as colony forming units per cubic meter of air (CFU/$m^3$). Bioaerosols were identified using standard microbial techniques by differential stains and/or microscopy. The environmental factors and possible causes of atopy based on ISAAC (International Study of Allergy and Asthma in Childhood) were collected by questionnaire. Results: The bioaerosol concentrations in indoor environments showed log-normal distribution (p < 0.01). Geometric mean (GM) and geometric standard deviation (GSD) of airborne bacteria and fungi in homes were 189.0 (2.5), 346.1(2.0) CFU/$m^3$, respectively. Indoor fungal levels were significantly higher than those of bacteria (p < 0.001). The concentration of airborne bacteria exceeded the limit recommended by the Korean Ministry of Environment, 800 CFU/$m^3$, in three out of 92 samples (3.3%) from 52 homes. The means of indoor to outdoor ratio (I/O) for airborne bacteria and fungi were 8.15 and 1.13, respectively. The source of airborne bacteria was not outdoors but indoors. GM of airborne bacteria and fungi were 217.6, 291.8 CFU/$m^3$ in the case's home and 162.0, 415.2 CFU/$m^3$ in the control's home respectively. The difference in fungal distributions between case and control were significant (p = 0.004) and the odds ratio was 0.996 (p = 0.027). Atopy was significantly associated with type of house (odds ratio = 1.723, p = 0.047) and income (odds ratio = 1.891, p = 0.041). Some of the potential allergic fungal genera isolated in homes were Cladosporium spp., Botrytis spp., Aspergillus spp., Penicillium spp., and Alternatia spp. Conclusions: These results suggest that there this should be either 'was little' meaning 'basically no significant association was found' or 'was a small negative' mean that an association was found but it was minor. It's a very improtant distinction. Association between airborne fungal concentrations and atopy and certain socioeconomic factors may affect the prevalence of childhood atopy.

• Korean Journal of Agricultural Science
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• v.49 no.4
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• pp.795-807
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• 2022

The development and commercialization of industrial genetically modified (GM) organisms is actively progressing worldwide, highlighting an increased need for improved safety management protocols. We sought to establish an environmental monitoring method, using real-time polymerase chain reaction (PCR) and propidium monoazide (PMA) treatment to develop a quantitative detection protocol for living GM microorganisms. We developed a duplex TaqMan quantitative PCR (qPCR) assay to simultaneously detect the selectable antibiotic gene, ampicillin (AmpR), and the single-copy Escherichia coli taxon-specific gene, D-1-deoxyxylulose 5-phosphate synthase (dxs), using a direct cell suspension culture. We identified viable engineered E. coli cells by performing qPCR on PMA-treated cells. The theoretical cell density (true copy numbers) calculated from mean quantification cycle (Cq) values of PMA-qPCR showed a bias of 7.71% from the colony-forming unit (CFU), which was within ±25% of the acceptance criteria of the European Network of GMO Laboratories (ENGL). PMA-qPCR to detect AmpR and dxs was highly sensitive and was able to detect target genes from a 10,000-fold (10^-4) diluted cell suspension, with a limit of detection at 95% confidence (LOD_95%) of 134 viable E. coli cells. Compared to DNA-based qPCR methods, the cell suspension direct PMA-qPCR analysis provides reliable results and is a quick and accurate method to monitor living GM E. coli cells that can potentially be released into the environment.

• Tuberculosis and Respiratory Diseases
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• v.64 no.1
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• pp.22-27
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• 2008

Background: Staphylococcus aureus frequently colonizes and infects hospitalized patients. Respiratory infections with Staphylococcus aureus are common in patients with compromised airway defenses. However the mechanisms of S. aureus invasion from colonization to the epithelium are unclear. Cell invasion by S. aureus would require destruction of the extracellular matrix, which is believed to be the result of increased matrix metalloproteinases (MMP) activity. Methods: In this study, respiratory epithelial cells were infected with S. aureus. After removing the extracellular bacteria by washing, the internalized bacteria in the cells were assessed by counting the colonized forming units (CFUs). The cell adhesion proteins, dysadherin and E-cadherin, were evaluated by Western blotting. The MMPs in the bacterial invasion were evaluated by pretreating the cells with GM6001, a MMP inhibitor. Results: The internalization of S. aureus was found to be both time and dose dependent, and the increase in MMP 2 and 9 activity was also dependent on the incubation time and the initial amount of bacterial inoculation. The invasion of S. aureus was attenuated by GM6001 after 12 hours incubation with a multiply of infection (MOI)=50. The expression of dysadherin, a membrane protein, was increased in a time and dose dependent manner, while the expression of E-cadherin was decreased. Conclusion: MMPs may mediate the invasion of S. aureus into epithelial cells.

• Journal of Environmental Health Sciences
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• v.42 no.6
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• pp.438-449
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• 2016

Objectives: Exposure to airborne bacteria is associated with adverse health effects such as respiratory and infectious diseases. This study evaluated airborne bacterial concentrations in the living rooms, kitchens, and toilets of 30 homes. Methods: Bacteria were sampled with an MAS100 impactor in three spaces in the subject homes between April 2014 and February 2015. Bacteria were grown on TSA plates for 48 hours at $35^{\circ}C$. The bacterial strains were isolated and amplified by polymerase chain reaction. Results: The most culturable bacteria were found in toilets ($624.0CFU/m^3$, GM: $417.3CFU/m^3$), followed by in the kitchen ($503.8CFU/m^3$, GM: $324.9CFU/m^3$). The dominant genera identified were: Staphylococcus sp.(19%), Micrococcus sp.(16%), and Bacillus sp.(11%) in the indoor air and Bacillus sp. (30%) in the outdoor air. Gram-positive bacteria comprised more than half of all colonies. Conclusion: In this study, culturable bacteria concentrations were higher than those reported in other spaces. Therefore, it is important to control relative humidity and remove moisture to prevent bacteria from multiplying. Additionally, the dominant species in indoor air were Staphylococcus sp. and Micrococcus sp. These are found on the human skin, mucous membranes, and hair, so human activity can affect bacterial distribution. Therefore, cleaning and controlling moisture are important for reducing indoor bacterial concentrations.

• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1401-1407
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• 2009

Many Lactobacillus strains have been promoted as good probiotics for the prevention and treatment of diseases. We engineered recombinant Lactobacillus casei, producing biologically active canine granulocyte macrophage colony stimulating factor (cGM-CSF), and investigated its possibility as a good probiotic agent for dogs. Expression of the cGM-CSF protein in the recombinant Lactobacillus was confirmed by SDS-PAGE and Western blotting methods. For the in vivo study, 18 Beagle puppies of 7 weeks of age were divided into three groups; the control group was fed only on a regular diet and the two treatment groups were fed on a diet supplemented with either $1\times10^9$ colony forming units (CFU)/day of L. casei or L. casei expressing cGM-CSF protein for 7 weeks. Body weight was measured, and fecal and blood samples were collected from the dogs during the experiment for the measurement of hematology, fecal immunoglobulin (Ig)A and IgG, circulating IgA and IgG, and canine corona virus (CCV)-specific IgG. There were no differences in body weights among the groups, but monocyte counts in hematology and serum IgA were higher in the group receiving L. casei expressing cGM-CSF than in the other two groups. After the administration of CCV vaccine, CCV-specific IgG in serum increased more in the group supplemented with L. casei expressing cGM-CSF than the other two groups. This study shows that a dietary L. casei expressing cGM-CSF enhances specific immune functions at both the mucosal and systemic levels in puppies.

• The Korea Journal of Herbology
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• v.27 no.4
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• pp.9-16
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• 2012

Objective : Canavalia gladiata DC semen (CGS) have been used to improve hematopoietic activity. In the current study, we investigated whether CGS regulate hemato-potentiating function using hematopoietic stem cells (HSCs) as a testing system. Methods : HSCs isolated from femur in mice with leukopenia and thrombocytopenia induced induced by CTX. Then, Real-time PCR was performed to measure the mRNA expression and hematopoietic related gene (EPO, IL-3, SCF, c-kit, GM-CSF), the phoaphorylation of GATA-1 and STAT-5a/b were observed by ELISA method, and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When HSCs were treated with CGS, the expression of hematopoietic related genes (EPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in HSCs. Additionally, CGS enhanced phosphorylation of STAT-1 and signal transducer and activator of transcription-5a/b (STAT-5a/b) in HSCs. Furthermore, CGS significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that CGS has hematopoietic enhancement via hematopoietic cytokine-mediated GATA-1/STAT-5a/b pathway.

• The Journal of Korean Medicine
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• v.23 no.1
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• pp.133-144
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• 2002

Objectives : Korean traditional medicinal herbs have been used to improve the function of the hematopoietic system. The purpose of this study is to examine the hematopoietic effects of Juakium-derivative on aplastic anemia. Methods : In vitro, after bone marrow cells of mice with aplastic anemia induced by benzene were cultured with Juakium-derivative, we measured the gene expressions of hematopoietic cytokines, the intracellular TPO and SCF expression and the colony number of CFU-GEMM and BFU-E. Results : The results are summarized as follows : 1. The gene expressions of hematopoietic cytokines (TPO, c-mpl, SCF, c-kit, IL-3, EPO, EpoR, GM-CSF) and the TPO and SCF expression of the group treated with Juakium-derivative increased significantly more than those of the control group. 2. The colony number of CFU-GEMM and BFU-E in the rEPO plus rIL-3 plus Juakium-derivative group increased significantly compared with the only rEPO plus rIL-3 group. Conclusion : It was acknowledged that Juakium-derivative has hemato-potentiating effects on aplastic anemia induced by benzene and it is expected that Juakium-derivative can be used clinically for the patient with hematopoietic system disorder. This study could also be used further as important research data for Korean oriental medicine about hemato-potentiating. The exact mechanism explaining our study would need to be studied further.