• Title/Summary/Keyword: Anaerobic culture

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Neuroprotective Effects of Medicinal Herbs in Organotypic Hippocampal Slice Cultures (뇌해마의 장기양 조직배양을 이용한 한약물의 뇌신경세포손상 보호효능 연구)

  • Jung, Hyuk-Sang;Sohn, Nak-Won;Lee, Won-Chul
    • The Journal of Internal Korean Medicine
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    • v.25 no.3
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    • pp.461-472
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    • 2004
  • Objectives : For the screening of neuroprotective effects of medicinal herbs, the complex system of animal models suffer some disadvantages in controlling critical parameters such as blood pressure and body temperature. Additionally, application of drugs to the appropriate brain area sometimes is difficult, due to poor permeability though the blood brain barrier, and so potential protective effects might be masked. Methods : Organotypic hippocampal slice culture (OHSC) method has the advantages of being relatively easy to prepare and of maintaining the general structure, including tissue integrity and the connections between cells. Drugs can easily be applied and neuronal damage can easily be quantified by using tissues and culture media. This study demonstrates neuroprotective effects of Puerariae radix (葛根, PR), Salviae miltiorrhizae radix (丹蔘, SR), Rhei rhizoma (大黃, RR), and Bupleuri radix (柴胡, BR). These were screenedand compared to MK-801, antagonist of NMDA receptors, by using OHSC of 1 week-old Sprague-Dawley rats. Oxygen/glucose deprivation (OGD) were conducted in an anaerobic chamber $(85%\;N_2,\;10%\;CO_2\;and\;5%\;H_2)$ in a deoxygenated glucose-free medium for 60 minutes. Water extracts of each herbs were treated to culture media with $5\;{\mu}g/ml$ for 48 hours. Results : Neuronal cell death in the cultures was monitored by densitometric measurements of the cellular uptake of propidium iodide (PI). PI fluorescence images were obtained at 48 hours after the OGD and medicinal herb treatment. Also TUNEL-positive cells in the CAI and DG regions and LDH concentrations in culture media were measured at 48 hours after the OGD. According to measured data, MK-801, PR, SR and BR demonstrated significant neuroprotective effect against excessive neuronal cell death and apoptosis induced by the OGD insult. Especially, PR revealed similar neuroprotective effect to MK-801 and RR demonstrated weak neuroprotective effect. Conclusions : These results suggest that OHSC can be a suitable method for screening of neuroprotective effects of medicinal herbs. (This work was supported by the research program of Dongguk University and Grant 01-PJ9-PG1-01CO03-0003 from Ministry of Health & Welfare.)

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Culture Conditions for Hydrogen Production of Enterobacter cloacae YJ-1 (수소생산을 위한 Enterobacter cloacae YJ-1의 배양조건)

  • Lee Ki-Seok;Kang Chang-Min;Chung Seon-Yong
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.446-450
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    • 2004
  • We investigated the effective culture conditions of anaerobic bacteria, Enterobacter cloacae YJ-1 on hydrogen production. It was cultured with 60 mL of working volume at $35^{\circ}C$, 120 rpm for 40 h. With culture time, hydrogen production and cell growth increased, but residual glucose and pH decreased. When the $2\%$ of glucose was used as single carbon source, hydrogen production was 975.1 mL/L. To enhance hydrogen productivity, mixed carbon sources of glucose and sucrose were added. The maximum hydrogen production was earned at the mixing ratio of 25:75, and it was 1319.5 mL/L. When we added 50 mM of phosphate to protect the pH drop in culture broth, hydrogen production increased 1.3 times more than that of initial concentration. The organic nitrogen sources were more effective than inorganic nitrogen for hydrogen production. Among organic nitrogen, yeast extract was the most effective and its hydrogen production was 1691.3 mL/L. Among 9 of mineral sources, Ferric citrate and $NaMoO_4$ were especially effective, and their productions were 1782.3 mL/L and 1784.8 mL/L, respectively.

Study on Low Temperature Tolerant Methane-Producing Bacteria for the Treatment of Agricultural and Livestock Wastes (농축산(農畜産) 폐기물(廢棄物) 처리(處理)를 위(爲)한 저온내성(低溫耐性) 메탄 생성균(生成菌)의 특성(特性)에 관(關)한 연구(硏究) II. 저온내성(低溫耐性) Clostridia 의 분리(分離))

  • Jung, Kwang-Yong;Kim, Jai-Joung;Daniels, Lacy
    • Korean Journal of Environmental Agriculture
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    • v.13 no.3
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    • pp.311-320
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    • 1994
  • This study was conducted to investigate the biochemical properties of isolated bacteria, low temperature tolerant methane-producing clostridia which were selected for using them as inoculum to anaerobic fermentation of agricultural and livestock wastes at low temperature. The results were; 1. Low temperature tolerant methane-producing clostridia were isolated from the samples which showed the high methanogenesis rate by enrichment culture at low temperature in cellulose medium. These clostridia, Clostridium botulinum SRC-64, Clostridium scatologens SRC-91 and Clostridium tyrobutyricum SRC-100, were isolated from swampy sediment at latitude $56.9^{\circ}N$, lake sediment IV at latitude $55.0^{\circ}N$, and tidal land soil II at latitude $37.0^{\circ}N$, respectively. The optimum growth temperature for these isolates was $37^{\circ}C$ and the minimum, around $10^{\circ}C$. They all had detectable amount of $F_{420}$, specific coenzyme of methanogens. 2. As anaerobic fermentation products of glucose SRC-64 produced $H_2$, acetic, isovaleric and caproic acid, SRC-91 produced $H_2$, propionic, butyric, valeric, and caproic acid, and SRC-100 produced only acetic and propionic acid. The isolates were produced $CH_4$ ranged from 2.6 to 8.68 n moles/ml for 2 days at $13^{\circ}C$.

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The Bacterial Community Structure in Biofilms of the RABC Process for Swine Butchery Wastewater Treatment (돼지 도축폐수 처리를 위한 RABC 공정의 생물막 세균군집 구조)

  • Sung, Gi-Moon;Lee, Dong-Geun;Park, Seong-Joo
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.56-65
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    • 2011
  • Culture-independent microscopic observations and 16S rDNA analyses were applied to describe the bacterial community inherent to the biofilm structure of the RABC (Rotating Activated Bacillus Contactors) process for swine butchery wastewater treatment. The ratios of Gram-positive bacterial counts to total bacterial counts of the RABC process were significantly increased in the last aeration tank as well as returned sludge, while those of the existing A2O (Anaerobic-Anoxic-Oxic) process maintained constant from aeration tanks to returned sludge. Totally nine phyla were recovered by 16S rDNA analysis, two of which were major groups: the Proteobacteria (64.1%) and the Actinobacteria (18.4%). The third major group was the endospore-forming Firmicutes (5.4%). The remaining six minor groups are the Bacteroidetes (3.3%), the Chlorobi (2.2%), the Nitrospirae (1.1%), the Chlorofleix (1.1%), the Acidobacteria (1.1%), and the Fusobacteria (1.1%). The ratio of endospore-forming bacteria was 19.4%, which was composed of the members of the Firmicutes phylum (5.4%) and the Intrasporangiaceae family (14.0%) of the Actinobacteria phylum. Nitrifying and denitrifying related- and phosphorus accumulating related-sequences were composed of 6.5% and 5.4% of total community, respectively, these could mean the high capacity of the RABC process to remove odor compounds and reduce eutrophication by efficient removing inorganic nutrients.

The Study of TCE Dechlorination using Geobacter lovleyi with Slow Release Substrate Applied (Slow Release Substrate를 이용한 Geobacter lovleyi의 TCE 탈염소화 연구)

  • Cha, Jae Hun;An, Sang Woo;Park, Jae Woo;Chang, Soon Woong
    • Journal of the Korean GEO-environmental Society
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    • v.13 no.9
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    • pp.53-59
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    • 2012
  • This study investigated characteristics of decomposition of tetrabutoxysilane (TBOS) as a slow release substrate (SRS) and on effect of TBOS decompostion compounds (acetate and butylate) for anaerobic dechlorination of trichloroethylene (TCE). In the batch experiment, TCE, cis-dichloroethene (cis-DCE), 1-butanol and TBOS were analysed by GC/FID and acetate and butylate were measured by HPLC. 1M of TBOS transferred and accumulated 4M of 1-butanol by abiotically hydrolysis reaction. The hydrolysis rate was in a range of 0.186 ${\mu}M/day$. On other hand, 1-butanol fermented to butyrate and acetate with indigenous culture from natural sediments. This results showed that TBOS could be used a slow release substrate in the natural sites. The dechlorinated potential of TCE with acetate and butyrate was increased with a decreasing initial TCE concentrations. In addition, first order coefficients of dechlorination with acetate as electron donor was higher then that with butyrate. It is because that dechlorination of Geobacter lovleyi was affected by substrate affinity, biodegradability and microbial acclimation on various substrates. However, dechlorinated potential of Geobacter lovleyi was decreased with accumulation cis-DCE in the anaerobic decholoronation process. The overall results indicated that SRS with Geobacter lovleyi might be a promising material for enhancing dechlorination of TCE on natural site and cis-DCE should be treated by ZVI as reductive material or by coexisting other dechlorinated bacteria.

Effects on Chemical Compositions and Digestibilities of the Bulking Agents as a Moisture Control and fermentation Methods of food Waste (음식찌꺼기의 발효사료화시 수분조절제와 발효방법이 화학적 조성분 및 소화율에 미치는 영향)

  • Bae, Dong-Ho
    • Journal of the Korea Organic Resources Recycling Association
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    • v.8 no.4
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    • pp.100-110
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    • 2000
  • Studies were conducted to know effects of the bulking agents (saw dusts, mushroom waste, wheat bran coconut meal, rice hulls) adding o moisture control, fermentation methods (aerobic and anaerobic) and periods (1 to 20 days) of food waste fermentation for animal feeds on chemical compositions and in vitro DDM (digestibility of dry matter). Experiment designs were focussed basically to obtain extension service data. The NDF (neutral detergent fiber) composition in the oak and pine saw dust were 93.5% and 95.4% (DM basis) in respectively. Thus, the fermented food waste feeds using saw dust (50%) increased NDF(12%), and decreased in vitro DDM(48%) compared to those of raw materials before aerobic fermentation. The oak saw dust showed higher DDM compared to pine. Mushroom wastes which is a residues of mushroom culture mixed originally willow saw dust (80%) and wheat bran (20%) showed quite higher feed value compared to both saw dusts. It was found that an in vitro DDM and NDF composition in fermented feeds appeared highly dependent or the NDF composition in bulking agents. With an increase wheat bran ratio substitute mushroom waste showed linearly decreased NDF, and increased in vitro DDM in the fermented food waste feeds. The fermented feeds added bottling agents composed higher NDF resulted in higher NDF and lower in vitro DDM with prolonged fermentation time. The feeds from anaerobic fermentation appeared lower NDF and higher in vitro DDM compared to those of aerobic fermentation.

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Bacterial Flora of the Intestine in Normal Captive Oriental White Storks (정상적인 사육 황새의 장내 세균총)

  • Han, Jae-Ik;Jang, Hye-Jin;Lee, Sook-Jin;Kang, Hyo-Min;Kim, Su-Kyung;Park, Shi-Ryoung;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.28 no.5
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    • pp.516-518
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    • 2011
  • A survey was conducted to examine the normal intestinal bacterial flora of captive Oriental white storks (Ciconia boyciana) maintained at the Korea Institute of Oriental White Stork Rehabilitation Research, Cheongwon, South Korea. From the cloaca of 44 healthy storks, 44 fecal samples were collected and cultured under aerobic and anaerobic conditions. The 16S ribosomal RNA gene sequences and the heat shock protein 60 gene were cloned and sequenced for bacterial identification. Under aerobic conditions, Enterococcus faecalis, Escherichia coli, Bacillus spp., Enterococcus avium, Enterococcus gallinarum, Pseudomonas spp., Alcaligenes spp., Enterobacter spp., Corynebacterium spp., and Proteus mirabilis were identified. Under anaerobic conditions, E. coli, Clostridium tertium, En. faecalis, and P. mirabilis were identified. E. coli, En. faecalis, or both were isolated from all samples. These results will add to the information available on this stork species and help for the interpretation of fecal culture results.

The Interactive Effect of These Bacterial Substrates on the Growth of Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis (Streptococcus gordonii, Fusobacterium nucleatum 및 Porphyromonas gingivalis의 상호작용이 성장에 미치는 영향)

  • Kim, A-Reum;Jeong, Moon-Jin;Ahn, Yong-Soon;Kim, Mi-Na;Kim, Sung-Im;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.15 no.2
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    • pp.209-219
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    • 2015
  • In order to explore an effect of interaction of Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis that are bacteria relevant to periodontal disease on its growth, the bacteria were incubated in trypticase soy hemin menadione broth at $37^{\circ}C$ $CO_2$ incubator for 7 days through anaerobic jar by single and co-culture with heat treated dead bacteria under anaerobic gas pack. In order to confirm growth level, absorbance was measured and for confirming colony structure and form, it was observed with scanning electron microscope. In order to confirm an effect on pathogenicity of P. gingivalis, real time reverse transcriptase polymerase chain reaction was implemented for expression analysis for rgpA gene that produces HRgpA which is gingipain. As a result, the following conclusion was obtained. Colony formation of S. gordonii and P. gingivalis was increased by other dead bacteria and in case of F. nucleatum, its colony formation was showed an aspect of being increased by dead bacterium of P. gingivalis but decreased by dead bacterium of S. gordonii. Therefore, it is considered that the strains being used for this study would affect interactively through bacterial cell itself as well as their interaction factor at the time of colony formation.

Correlation between Characteristics of SOD in Coastal Sewage and Predictive Factor (연안 저질 SOD의 특성과 유발 영향인자에 대한 상관관계)

  • Kim, Beom-Geun;Khirul, Md Akhte;Kwon, Sung-Hyun;Cho, Dae-Chul
    • Korean Journal of Environment and Ecology
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    • v.33 no.5
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    • pp.596-604
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    • 2019
  • This study conducted a sediment culture experiment to investigate the effects of sediment oxygen demand (SOD) and environmental factors on sediment and water quality. We installed a leaching tank in the laboratory, cultured it for 20 days, and analyzed the relationship between P and Fe in the sediment. As a result, the dissolved oxygen of the water layer decreased with time, while the oxidation-reduction potential of the sediment progressed in the negative direction to form an anaerobic reducing environment. The SOD was measured to be 0.05 mg/g at the initial stage of cultivation and increased to 0.09 mg/g on the 20th day, indicating the tendency of increasing consumption of oxygen by the sediment. The change is likely to have caused by oxygen consumption from biological-SOD, which is the decomposition of organic matter accumulated on the sediment surface due to the increase of chl-a, and chemical-SOD consumed when the metal-reducing product produced by the reduction reaction is reoxidized. The correlation between SOD and causality for sediment-extracted sediments was positive for Ex-P and Org-P and negative for Fe-P. The analysis of the microbial community in the sediment on the 20th day showed that anaerobic iron-reducing bacteria (FeRB) were the dominant species. Therefore, when the phosphate bonded to the iron oxide is separated by the reduction reaction, the phosphate is eluted into the water to increase the primary productivity. The reduced substance is reoxidized and contributes to the oxygen consumption of the sediment. The results of this study would be useful as the reference information to improve oxygen resin.

Microenvironments and Cellular Proliferation Affected by Oxygen Concentration in Non-Small Cell Lung Cancer Cell Line (비소세포폐암주에서 산소 농도에 따른 미세 배양 환경과 세포 증식능)

  • Shin, Jong Wook;Jeon, Eun Ju;Kwak, Hee Won;Song, Ju Han;Lee, Young Woo;Jeong, Jae Woo;Choi, Jae Cheol;Kim, Jae-Yeol;Park, In Won;Choi, Byoung Whui
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.3
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    • pp.242-250
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    • 2007
  • Background: Abnormal angiogenesis can induce hypoxia within a highly proliferating tumor mass, and these hypoxic conditions can in turn create clinical problems, such as resistance to chemotherapy. However, the mechanism by which hypoxia induces these changes has not yet been determined. Therefore, this study was conducted to determine how hypoxia induces changes in cell viability and extracellular microenvironments in an in vitro culture system using non-small cell lung cancer cells. Methods: The non-small cell lung cancer cell line, A549 was cultured in DMEM or RPMI-1640 media that contained fetal bovine serum. A decrease in the oxygen tension of the media that contained the culture was then induced in a hypoxia microchamber using a $CO_2-N_2$ gas mixture. A gas analysis and an MTT assay were then conducted. Results: (1) The decrease in oxygen tension was checked the anaerobic gas mixture for 30 min and then reoxygenation was induced by adding a 5% $CO_2-room$ air gas mixture to the chamber. (2) Purging with the anaerobic gas mixture was found to decrease the further oxygen tension of cell culture media. (3) The low oxygen tension resulted in a low pH, lactic acidosis and a decreased glucose concentration in the media. (4) The decrease in glucose concentration that was observed as a result of hypoxia was markedly different when different types of media were evaluated. (5) The decrease in oxygen tension inhibited proliferation of A549 cells. Conclusion: These data suggests that tumor hypoxia is associated with acidosis and hypoglycemia, which have been implicated in the development of resistance to chemotherapy and radiotherapy.