• Title/Summary/Keyword: Dynamic cell culture

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Investigation of Nanofiber and Thermosensitive Scaffold for Intervertebral Disc through Organ Culture (기관배양을 통한 추간판 재생용 나노파이버 및 온도 감응성 지지체에 대한 검증)

  • Lee, Yong-Jae;Shin, Ji-Won;Shin, Ho-Jun;Kim, Chan-Hwan;Park, Ki-Dong;Bae, Jin-Woo;Seo, Hyoung-Yeon;Kim, Young-Jick;Shin, Jung-Woog
    • Journal of Biomedical Engineering Research
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    • v.28 no.4
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    • pp.512-519
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    • 2007
  • The purpose of this study is to investigate the potential of a novel tissue engineering approach to regenerate intervertebral disc. In this study, thermosensitive scaffold (chitosan-Pluronic hydrogel) and nanofiber were used to replace the nucleus pulposus (NP) and annulus fibrosus of a degenerated intervertebral disc, leading to an eventual regeneration of the disc using the minimally invasive surgical procedure and organ culture. In preliminary study, disc cells were seeded into the scaffolds and cellular responses were assessed by MTT assay and scanning electron microscopy (SEM). Based on these results, we could know that tissue engineered scaffolds might provide favorable environments for the regeneration of tissues. Organ culture was performed in fresh porcine spinal motion segments with endplates on both sides. These spinal motion segments were classified into three groups: control (Intact), injured NP (Defect), and inserting tissue engineered scaffolds (Insert). The specimens were cultivated for 7 days, subsequently structural stability, cell proliferation and morphological changes were evaluated by the relaxation time, quantity of DNA, GAG and histological examination. In these results, inserting group showed higher relaxation time, reduced decrement of DNA contents, and accumulated GAG amount. Consequently, the tissue engineered scaffolds used in this study seen to be a promising base scaffolds for regenerative intervertebral disc due to its capacity to absorb external dynamic loading and the possible ideal environment provided for disc cell growing.

Local protein synthesis in neuronal axons: why and how we study

  • Kim, Eunjin;Jung, Hosung
    • BMB Reports
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    • v.48 no.3
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    • pp.139-146
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    • 2015
  • Adaptive brain function and synaptic plasticity rely on dynamic regulation of local proteome. One way for the neuron to introduce new proteins to the axon terminal is to transport those from the cell body, which had long been thought as the only source of axonal proteins. Another way, which is the topic of this review, is synthesizing proteins on site by local mRNA translation. Recent evidence indicates that the axon stores a reservoir of translationally silent mRNAs and regulates their expression solely by translational control. Different stimuli to axons, such as guidance cues, growth factors, and nerve injury, promote translation of selective mRNAs, a process required for the axon's ability to respond to these cues. One of the critical questions in the field of axonal protein synthesis is how mRNA-specific local translation is regulated by extracellular cues. Here, we review current experimental techniques that can be used to answer this question. Furthermore, we discuss how new technologies can help us understand what biological processes are regulated by axonal protein synthesis in vivo.

Analysis of the Bacterial Composition During Kochujang, a Korean Traditional Fermented Hot Pepper-soybean Paste, Fermentation

  • Park, Sun-Jung;Chang, Jin-Hee;Cha, Seong-Kwan;Moon, Gi-Seong
    • Food Science and Biotechnology
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    • v.18 no.4
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    • pp.1035-1037
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    • 2009
  • In this study we analyzed the dynamic changes in microbiota composition during kochujang fermentation at $30^{\circ}C$. During fermentation, the viable cell counts slowly increased and reached $3.2{\times}10^7$ for aerobic bacteria, $8.3{\times}10^3$ for yeast, and $1.4{\times}10^3$ CFU/mL for fungi after 60 days. Bacilli were found to be the most dominant microorganisms throughout the fermentation process. Using the culture dependent method Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloquefaciens were found to be the main species during the early stages of fermentation; however, Bacillus pumilus and Bacillus stearothermophilus became the most dominant species during the late stage of fermentation. In contrast, when the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method was used Bacillus ehimensis was found to be the dominant species during the early stage of fermentation and Bacillus megaterium, B. pumilus, B. subtilis, and B. licheniformis were dominant in the ate stages. These results indicate various other Bacillus species rather than just B. subtilis and B. licheniformis might be involved in the fermentation of kochujang.

The Interkingdom Interaction with Staphylococcus Influences the Antifungal Susceptibility of the Cutaneous Fungus Malassezia

  • Juan Yang;Sungmin Park;Hyun Ju Kim;Sang Jun Lee;Won Hee Jung
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.180-187
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    • 2023
  • The skin is a dynamic ecosystem on which diverse microbes reside. The interkingdom interaction between microbial species in the skin microbiota is thought to influence the health and disease of the skin although the roles of the intra- and interkingdom interactions remain to be elucidated. In this context, the interactions between Malassezia and Staphylococcus, the most dominant microorganisms in the skin microbiota, have gained attention. This study investigated how the interaction between Malassezia and Staphylococcus affected the antifungal susceptibility of the fungus to the azole antifungal drug ketoconazole. The susceptibility was significantly decreased when Malassezia was co-cultured with Staphylococcus. We found that acidification of the environment by organic acids produced by Staphylococcus influenced the decrease of the ketoconazole susceptibility of M. restricta in the co-culturing condition. Furthermore, our data demonstrated that the significant increased ergosterol content and cell membrane and wall thickness of the M. restricta cells grown in the acidic environment may be the main cause of the altered azole susceptibility of the fungus. Overall, our study suggests that the interaction between Malassezia and Staphylococcus influences the antifungal susceptibility of the fungus and that pH has a critical role in the polymicrobial interaction in the skin environment.

Fermentative Water Purification based on Bio-hydrogen (생물학적 수소 발효를 통한 수처리 시스템)

  • Lee, Jung-Yeol;Chen, Xue-Jiao;Min, Kyung-Sok
    • Journal of Korean Society on Water Environment
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    • v.27 no.6
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    • pp.926-931
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    • 2011
  • Among various techniques for hydrogen production from organic wastewater, a dark fermentation is considered to be the most feasible process due to the rapid hydrogen production rate. However, the main drawback of it is the low hydrogen production yield due to intermediate products such as organic acids. To improve the hydrogen production yield, a co-culture system of dark and photo fermentation bacteria was applied to this research. The maximum specific growth rate of R. sphaeroides was determined to be $2.93h^{-1}$ when acetic acid was used as a carbon source. It was quite high compared to that of using a mixture of volatile fatty acids (VFAs). Acetic acid was the most attractive to the cell growth of R. sphaeroides, however, not less efficient in the hydrogen production. In the co-culture system with glucose, hydrogen could be steadily produced without any lag-phase. There were distinguishable inflection points in the accumulation of hydrogen production graph that resulted from the dynamic production of VFAs or consumption of it by the interaction between the dark and photo fermentation bacteria. Lastly, the hydrogen production rate of a repeated fed-batch run was $15.9mL-H_2/L/h$, which was achievable in the sustainable hydrogen production.

New surveillance concepts in food safety in meat producing animals: the advantage of high throughput 'omics' technologies - A review

  • Pfaffl, Michael W.;Riedmaier-Sprenzel, Irmgard
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.7
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    • pp.1062-1071
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    • 2018
  • The misuse of anabolic hormones or illegal drugs is a ubiquitous problem in animal husbandry and in food safety. The ban on growth promotants in food producing animals in the European Union is well controlled. However, application regimens that are difficult to detect persist, including newly designed anabolic drugs and complex hormone cocktails. Therefore identification of molecular endogenous biomarkers which are based on the physiological response after the illicit treatment has become a focus of detection methods. The analysis of the 'transcriptome' has been shown to have promise to discover the misuse of anabolic drugs, by indirect detection of their pharmacological action in organs or selected tissues. Various studies have measured gene expression changes after illegal drug or hormone application. So-called transcriptomic biomarkers were quantified at the mRNA and/or microRNA level by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) technology or by more modern 'omics' and high throughput technologies including RNA-sequencing (RNA-Seq). With the addition of advanced bioinformatical approaches such as hierarchical clustering analysis or dynamic principal components analysis, a valid 'biomarker signature' can be established to discriminate between treated and untreated individuals. It has been shown in numerous animal and cell culture studies, that identification of treated animals is possible via our transcriptional biomarker approach. The high throughput sequencing approach is also capable of discovering new biomarker candidates and, in combination with quantitative RT-qPCR, validation and confirmation of biomarkers has been possible. These results from animal production and food safety studies demonstrate that analysis of the transcriptome has high potential as a new screening method using transcriptional 'biomarker signatures' based on the physiological response triggered by illegal substances.

Chemical signalling within the rumen microbiome

  • Katie Lawther;Fernanda Godoy Santos;Linda B Oyama;Sharon A Huws
    • Animal Bioscience
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    • v.37 no.2_spc
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    • pp.337-345
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    • 2024
  • Ruminants possess a specialized four-compartment forestomach, consisting of the reticulum, rumen, omasum, and abomasum. The rumen, the primary fermentative chamber, harbours a dynamic ecosystem comprising bacteria, protozoa, fungi, archaea, and bacteriophages. These microorganisms engage in diverse ecological interactions within the rumen microbiome, primarily benefiting the host animal by deriving energy from plant material breakdown. These interactions encompass symbiosis, such as mutualism and commensalism, as well as parasitism, predation, and competition. These ecological interactions are dependent on many factors, including the production of diverse molecules, such as those involved in quorum sensing (QS). QS is a density-dependent signalling mechanism involving the release of autoinducer (AIs) compounds, when cell density increases AIs bind to receptors causing the altered expression of certain genes. These AIs are classified as mainly being N-acyl-homoserine lactones (AHL; commonly used by Gram-negative bacteria) or autoinducer-2 based systems (AI-2; used by Gram-positive and Gram-negative bacteria); although other less common AI systems exist. Most of our understanding of QS at a gene-level comes from pure culture in vitro studies using bacterial pathogens, with much being unknown on a commensal bacterial and ecosystem level, especially in the context of the rumen microbiome. A small number of studies have explored QS in the rumen using 'omic' technologies, revealing a prevalence of AI-2 QS systems among rumen bacteria. Nevertheless, the implications of these signalling systems on gene regulation, rumen ecology, and ruminant characteristics are largely uncharted territory. Metatranscriptome data tracking the colonization of perennial ryegrass by rumen microbes suggest that these chemicals may influence transitions in bacterial diversity during colonization. The likelihood of undiscovered chemicals within the rumen microbial arsenal is high, with the identified chemicals representing only the tip of the iceberg. A comprehensive grasp of rumen microbial chemical signalling is crucial for addressing the challenges of food security and climate targets.

The Beneficial Effects of Pectin on Obesity In vitro and In vivo (In vitro 및 In vivo에서 펙틴의 비만 억제 효과)

  • Kwon, Jin-Young;Ann, In-Sook;Park, Kun-Young;Cheigh, Hong-Sik;Song, Yeong-Ok
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.1
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    • pp.13-20
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    • 2005
  • The effects of pectin on obesity was studied using 3T3-L1 pre-adipocytes and rats fed 20% high fat diets. The concentration of leptin released from 3T3-L1 adipocytes in the presence of pectin was significantly decreased by 85% compared to that of the control (p<0.05), however, glycerol concentration was not changed. These data indicate that pectin seems to inhibit lipids accumulation in the adipocytes rather than enhance the lipolytic activity. Forty Sprague Dawley rats were fed 20% high fat diet for 8 weeks to induce obesity and then divided equally into four groups. Experimental groups were normal diet group (ND), high fat diet group (HFD), HDF with 10% pectin group (HFP10), and HDF with 20% pectin group (HFP20). Diet for the each group was prepared to be iso-caloric following AIN-76 guideline. After obesity was induced, rats were placed on an restricted diet for 9 weeks. The body weight of HFD increased 50% (p<0.05) compared to the ND, while it was decreased by 12% and 16% for HFP10 and HFP20, respectively (p<0.05). The relative amount of visceral fats for HFDl0 and HFD20 were decreased by 45% and 59% compared to that of HDF (130%), respectively (p<0.05). Pectin seems to have a greater effect on reducing visceral fats accumulation than weight reduction. Significantly increased level of triglyceride, total cholesterol or LDL-cholesterol in the plasma of HFD was returned to the normal or even below the normal by pectin diet, while the level of HDL-cholesterol increased. Lipid lowering effect was also observed in the liver and heart. These effects of pectin were dosedependent. In conclusion, the beneficial effect of pectin on the obesity was observed from cell culture experiment and animal study in terms of inhibiting the accumulation of lipids in the adipocytes.