• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1583-1588
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• 2015

The large intestine is a home for trillions of microbiota, which confer many benefits on the host, including production of vitamins, absorption of nutrients, pathogen displacement, and development of the immune system. For several decades, germ-free animals have been used to study the interaction between the host and its microbiota. This minireview describes the technical aspects for establishing and maintaining germ-free animals and highlights the advantages and disadvantages for germ-free animals as experimental models.

• Journal of The Korea Institute of Healthcare Architecture
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• v.12 no.3
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• pp.35-41
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• 2006

At the moment, a lot of interest in the research on Gnotobiotic Pigs are increasing in order to produce alternative human organs. So, it is very important to design and build proper housing facilities for Germ Free Pigs. Among the design issues related to Gnotobiotic Pigs' farms, circulation system takes a high position because it carries an important role in keeping the pig's housing environment aseptic. Considering those, this study aims to propose the guidelines for the design of circulation system in Germ Free Pig's facilities. The results of this study are as follows. At first, functional areas of Germ Free Pigs' facilities have been divided into three categories according to the clean level; aseptic area, semi-aseptic area, and non-aseptic area. Secondly, the basic principles of circulation system have been proposed. Finally, circulation system of Gnotobiotic Pigs' facility has been explored as a form of diagram according to the circulating subjects. These include human circulation, pig's circulation, and goods' circulation. This study has some limitations in that it is transcendent and lacks empirical evidence. Despite of some weaknesses, it is expected to give some useful guidelines for the design of circulation system in Germ Free Pigs' facilities.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.7
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• pp.958-963
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• 2006

The effect of ginsenosides (GS) on germ cell proliferation was evaluated with a chicken ovarian germ-somatic cell coculture model and the mechanism involving protein kinase C (PKC) pathway was investigated. Ovarian cells were cultured in serum-free McCoy's 5A medium and challenged with GS alone or in combinations with PKC activator (phorbol 12-myristate 13-acetate, PMA) or inhibitor ($H_7$) for 48 h. The number of germ cells was counted and the proliferating cells were identified by immunocytochemistry of proliferating cell nuclear antigen (PCNA). Results showed that GS significantly increased germ cell proliferation and this stimulating effect was further increased by PMA, but inhibited by H7, in a dose-dependent manner. Moreover, GS-elevated PCNA expression and the PCNA -labeling index of germ cells displayed similar changes with the increased numbers of germ cells. These results indicated that GS stimulated proliferation of ovarian germ cells with involvement of the PKC-mediated system.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.10
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• pp.1364-1368
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• 2010

Gnotobiotic piglets (n = 10) were hand-reared in conventional germ-free facilities. Piglet body temperatures were measured with rectal and non-contact infrared thermometry (NIFT) on the lower eyelid, auricular center and margin, parietal regions, axilla, central abdomen and dorsum, and the perianal region. Body temperature measurements at central abdomen, cranial dorsum, and perianal regions had NIFT values which had a significant linear relationship (p<0.0001) with rectal thermometry. The predicted equations of between-subject formulas were calculated as follows: rectal temperature, 28.07489+0.30372${\times}$central abdominal surface temperature; rectal temperature, 34.02799+0.15197${\times}$central dorsum surface temperature; and rectal temperature, 33.87937+0.15676${\times}$perianal temperature. These results suggested that NIFT could serve as a valid alternative to rectal thermometry in a portable germ-free facility without disturbing experimental animals. The development of a NIFT body temperature evaluation that does not require animal restraint is clinically advantageous, particularly in gnotobiotic piglets, and would be significantly less stressful for experimental procedures in germ-free facilities.

• Molecules and Cells
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• v.37 no.6
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• pp.473-479
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• 2014

Spermatogonial stem cells (SSCs, also called germline stem cells) are self-renewing unipotent stem cells that produce differentiating germ cells in the testis. SSCs can be isolated from the testis and cultured in vitro for long-term periods in the presence of feeder cells (often mouse embryonic fibroblasts). However, the maintenance of SSC feeder culture systems is tedious because preparation of feeder cells is needed at each subculture. In this study, we developed a Matrigel-based feeder-free culture system for long-term propagation of SSCs. Although several in vitro SSC culture systems without feeder cells have been previously described, our Matrigel-based feeder-free culture system is time- and cost-effective, and preserves self-renewability of SSCs. In addition, the growth rate of SSCs cultured using our newly developed system is equivalent to that in feeder cultures. We confirmed that the feeder-free cultured SSCs expressed germ cell markers both at the mRNA and protein levels. Furthermore, the functionality of feeder-free cultured SSCs was confirmed by their transplantation into germ cell-depleted mice. These results suggest that our newly developed feeder-free culture system provides a simple approach to maintaining SSCs in vitro and studying the basic biology of SSCs, including determination of their fate.

• Natural Product Sciences
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• v.8 no.2
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• pp.35-43
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• 2002

Glycosides of herbal medicines, such as glycyrrhizin, ginsenosides, kalopanaxsaponins, rutin and ponicirin, were studied regarding their metabolic fates and pharmacological actions in relation to intestinal bacteria using germ-free, gnotobiotic and conventional animals. When glycyrrhizin (GL) was orally administered, $18{\beta}-glycyrrhetinic\;acid\;(GA)$, not GL, was detected in plasma and intestinal contents of gnotobiotic and conventional rats. However, GA could not be detected in germ-free rats. When GL was incubated with human intestinal bacteria, it was directly metabolized to GA (>95%) or via $18{\beta}-glycyrrhetinic\;acid-3-{\beta}-D-glucuronide$(>5%). Orally administered GL was effective in gnotobiotic and conventional rats for liver injury induced by carbon tetrachloride, but was not effective in germ-free rats. When ginseng saponins were orally administered to human beings, compound K in the plasma was detected, but the other protopanxadiol saponins were not detected. The compound K was active for tumor metastasis and allergy. When kalopanaxsaponins were incubated with human intestinal microflora, they were metabolized to kalopanaxsaponin A, kalopanaxsaponin I and hederagenin. These metabolites were active for rheumatoid arthritis and diabetic mellitus while the other kalopanxsaponins were not. When flavonoid glycosides were orally administered to animals, aglycones and/or phenolic acids were detected in the urine. The metabolic pathways proceeded by intestinal bacteria rather than by liver or blood enzymes. These metabolites, aglycones and phenolic acids, showed antitumor, antiinflammatory and antiplatelet aggregation activities. These findings suggest that glycosides of herbal medicines are prodrugs.

• Journal of The Korea Institute of Healthcare Architecture
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• v.14 no.4
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• pp.39-46
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• 2008

Construction and operation of Germ Free Pigs' facilities are very expensive because pigs' rooms and other major rooms of the facility require germfree environments. Especially, running the HVAC system of aseptic facilities requires a lot of expenses. However, proper location and efficient shape of outlets/inlets for the ventilation of the room can reduce the excessive running cost. In order to do that, this study proposes alternative location and shape of ventilation outlets/inlets to the existing design pattern in germfree pigs' room. The design condition of this study is the maintenance of adequate temperature(24$^{\circ}C$, $NH_3$concentration level(below 1.5 ppm), and air stream speed(below .25m/sec) in the pigs' room for the summer and the winter together. As the Software Program, FLUENT(Ver. 6.2) has been used for the analysis of proposed ventilation patterns. In conclusion, wall inlets and ceiling inlet/outlet are advisable in summer, wall inlets and ceiling outlets is advisable in winter. As far as the shape is concerned, diffuser type for the ceiling outlet is desirable.

• Development and Reproduction
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• v.26 no.4
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• pp.145-153
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• 2022

The gut microbiota is involved in the maintenance of physiological homeostasis and is now recognized as a regulator of many diseases. Although germ-free mouse models are the standard for microbiome studies, mice with antibiotic-induced sterile intestines are often chosen as a fast and inexpensive alternative. Pathophysiological changes in the gut microbiome have been demonstrated, but there are no reports so far on how such alterations affect the bacterial composition of the uterus. Here we examined changes in uterine microbiota as a result of gut microbiome disruption in an antibiotics-based sterile-uterus mouse model. Sterility was induced in 6-week-old female mice by administration of a combination of antibiotics, and amplicons of a bacteria marker gene (16S rRNA) were sequenced to decipher bacterial community structures in the uterus. At the phylum-level, Proteobacteria, Firmicutes, and Actinobacteria were found to be dominant, while Ralstonia, Escherichia, and Prauserella were the major genera. Quantitative comparisons of the microbial contents of an antibiotic-fed and a control group revealed that the treatment resulted in the reduction of bacterial population density. Although there was no significant difference in bacterial community structures between the two animal groups, β-diversity analysis showed a converged profile of uterus microbiotain the germ-free model. These findings suggest that the induction of sterility does not result in changes in the levels of specific taxa but in a reduction of individual variations in the mouse uterus microbiota, accompanied by a decrease in overall bacterial population density.

• Korean Journal of Food Science and Technology
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• v.51 no.2
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• pp.97-102
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• 2019

Wheat germ, which is rich in nutrients and phytochemicals, is a by-product during the milling process of wheat kernel. In this study, we aimed to increase the amount of bioactive 2,6-dimethoxy-1,4-benzoquinone (2,6-DMBQ) in wheat germ using the cell-wall-degrading enzyme cellulase (Celluclast 1.5L). The amounts of organic acids, free sugars, and 2,6-DMBQ in wheat germ treated with Celluclast 1.5L were evaluated at various reaction times and temperatures. The results of reversed-phase high-performance liquid chromatography of Celluclast 1.5L-treated wheat germ revealed 2,6-DMBQ, four organic acids (tartaric, acetic, lactic, and succinic acids), and three free sugars (sucrose, fructose, and glucose). As reaction time and temperature of the mixture of wheat germ and Celluclast 1.5L increased, the contents of four organic acids, glucose, fructose, and 2,6-DMBQ increased, but that of sucrose decreased. Taken together, these results suggest that Celluclast 1.5L-treated wheat germ containing increased amounts of 2,6-DMBQ serves as a source of functional ingredients in food industry.

• Korean Journal of Food Science and Technology
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• v.27 no.4
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• pp.478-482
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• 1995

The changes of the lipid composition and of the contents of carotenoid and tocopherol in wheat germ were studied during the storage at $30^{\circ}C$. The contents of triglyceride and free fatty acid were changed from 66% and 7% to 49% and 24% respectively after 30 days. The predominant free fatty acids were lauric acid (29%), palmitic acid (21%) and linoleic acid (20%), however, linoleic acid increased to 30%, lauric acid reduced to 21% after storage of 30 days. The carotenoids in the wheat germ were ${\beta}-carotene,\;{\alpha}-carotene$, lutein and taraxanthin, and the contents of these were 306, 59, 383 and 356 ng/g wheat germ, respectively. Their contents, however, were reduced to 36, 4, 203 and 149 ng respectively after 20 storage days. Especially, degradation rate of ${\beta}-carotene$ was 22.5 ng/day. The tocopherol isomers in wheat germ were ${\alpha}-,\;{\beta}-\;and\;{\gamma}-tocopherol$, and they reduced from $55,\;48\;and\;38\;{\mu}g/g$ wheat germ to 35, 32 and $32\;{\mu}g$ respectively after 20 storage days. The ${\alpha}-tocopherol$ was degraded by $1.26\;{\mu}g/day$ at this storage condition.