• 생명과학회지
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• 제15권4호
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• pp.626-632
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• 2005

폴리아민은 모든 진핵세포에서 발견되는 다가 양이온성의 저분자 물질이며 세포성장에 필수적인 것으로 알려져 있다. 본 논문에서는 폴리아민의 역할 중에서 산화적인 스트레스에 대한 세포보호 효과를 연구하였다. 쥐의 간세포주인 $Ac_2F$에 산화 스트레스를 유발하기 위하여 2,2'-azobis(2-amidinopropane)dehydrochloride (AAPH)를 처리하였을 때, 세포증식은 농도 의존적으로 감소하였다. 배지에 폴리아민을 첨가하였을 때 세포성장은 농도 의존적으로 증가하였으며 ROS 발생은 현저히 감소하였다. 폴리아민 가운데 특히 spermidine과 spermine이 뚜렷한 세포증식효과를 보였다. Spermine의 경우, $20{\mu}M$농도에서 AAPH에 의해 유도된 ROS발생을 $45\%$나 감소시켰다. 산화 스트레스에 관여하는 효소들 가운데 주된 효소인 superoxide dismutate (SOD)와 catalase (CAT)의 세포 내 단백질을 Western blotting으로 조사한 결과, AAPH는 이 두 가지 단백질의 생성을 억제한 것으로 나타났다. 그러나 spermine을 처리하였을 때 두 단백질의 생산은 모두 정상적으로 회복이 되었다. 또한 세포주기의 중요한 조절 단백질인 cyclin E 역시 AAPH에 의하여 생성이 억제되었다. 이는 AAPH에 의하여 생성된 ROS가 세포주기의 S phase의 진행을 억제한 것으로 생각된다. AAPH에 의한 cyclin E의 억제는 spermine에 의하여 정상적으로 회복되었다. 위와 같은 Spermine의 항산화 효과는 ethidium bromide와 acridine orange를 이용하여 형태학적으로도 증명되었다.

• 대한약침학회지
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• 제7권1호
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• pp.5-14
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• 2004

Acupuncture has fairly good weight-reducing effect in treating simple obesity due to the neuroendocrine regulation. In this study, the antiobesity effects of herbal acupuncture(HA) with Sang-hwang(phellinus linteus) at Fuai(SP16) were investigated in the rat fed on high-fat(HF) diet. Sang-hwang mushroom has been proven to have anti-carcinogenic effects and Sang-hwang extracts are highly effective in treatment and preventive treatment of AIDS, diabetes and high blood-pressure. To determine whether the Sang-hwang herbal acupuncture may have the anti-obesity effect, male Sprague-Dawley(4-wk-old) rats were fed a HF diet for 5 wk, which produced significant weight gain compared to rats were fed a normal diet, and then herbal acupuncture were treated for 3 wk in HF diet group. The body weight, food consumption, food effeciency ratio(FER), body fat mass, plasma nitric oxide(NO) were investigated in rats fed on normal diet, HF diet, and HF diet with HA(HF-diet-HA) groups. NO has been proposed to be involved in the regulation of food intake. In addition, the expression of appetite peptides such as orexigenic peptide neuropeptide Y(NPY) and the anorectic peptide cholecystokinin(CCK) were observed in the hypothalamus. HF-HA group reduced body weight gain, FER, body fat contents and NO concentration compared to HF diet group. The expression of NPY was reduced in arcuate nucleus(ARC), and CCK was increased in the paraventricular nucleus(PVN) after treatment of HA. In conclusion, Sang-hwang HA reduced adipocity, plasma NO and hypothalamic NPY, but increased CCK expression in the HF dietinduced obesity rat, therefore HA may have anti-obesity action through regulating body weight and appetite peptide of the central nervous system.

• Applied Microscopy
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• 제28권2호
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• pp.171-180
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• 1998

본 연구에서는 dehydrocholic acid를 투여한 흰쥐 간세포의 vacuolar apparatus를 세포화학적으로 관찰하고자 하였다. 박절편에서 Golgi 장치의 형성면 수조는 소낭상으로 팽대된 부분이 열지어 있었으며 어떤것은 돌출되어 있었다. 이러한 소낭 모양의 돌출부는 용해소체 표면에서도 관찰되었다. 소포들은 Golgi 장치의 형성면 수조와 용해소채 및 담세관 주위에서 볼 수 있었으며 소포가 담세관에 융합되어 있는 것도 관찰되었다. 이러한 소견들은 dehydrocholic acid 투여 후 20분이 경과된 흰쥐에서 현저하였다. 정상군이나 실험군에서 거의 모든 Golgi 장치의 형성면은 담세관을 향하고 있었다. 박절편에서 Golgi 장치의 형성면 수조와 소낭 모양의 돌출부 및 소포는 가시물질을 거의 함유하고 있지 않았다. Osmium은 이들구조에서 심하게 침착되어 있었다. 용해소체와 그 주위에 있는 소포에서는 acid phosphatase 활성이 나타났다. 그러나 담세관 주위에 있는 소포에서는 osmium의 침착이나 acid phosphatase 활성이 경하게 나타나거나 관찰되지 않았다. 이러한 증거들로 미루어 소포는 Golgi 장치의 형성면과 용해소체에서 유래되며 이들소포가 담세관에 융합됨으로서 담즙산이 분비될 것으로 생각된다. 그러나 소포가 담세관에 접근해 감에 따라 소포의 효소활성은 저하되는 것 같이 보인다.

• 한국수정란이식학회지
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• 제24권4호
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• pp.265-269
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• 2009

The purpose of this study was to determine the effect of different feeding ratios of whole crop barley silage on the embryo production in Hanwoo donors. All donors were basically fed 2.5 kg concentrate daily. Donors were divided into three groups according to the different feeding of forage; hay 70% and rice straw 30% (control, n = 21), whole crop barley silage 80% and rice straw 20% (T1, n = 25), and whole crop barley silage 60% and rice straw 40% (T2, n = 23) fed based on TDN 6.70/ BW 500 kg. All Hanwoo donors received a CIDR together with injections of 1 mg estradiol benzoate and 50 mg progesterone ($P_4$, Day 0). Four days later, they were superovulated with 28 mg FSH twice daily IM in decreasing doses over 4 days. Then donors received 2 doses of $PGF_2{\alpha}$ (25 and 15 mg) with the 5th and 6th injections of FSH on Day 6. CIDR were withdrawn at the 6th FSH injection and the donors received $100\;{\mu}g$ GnRH 36 h after the second $PGF_2{\alpha}$ injection. The donors were artificially inseminated twice, at 8 and 24 h after GnRH, and embryos were recovered 7 or 8 days after the 1st insemination. The flush rate of the donors following positive superovulation responses did not differ among groups (76.2~96.0%, p>0.05). The number of corpus luteum (CL) at embryo recovery also did not differ among groups (10.6~14.0, p>0.05). Furthermore, the mean numbers of total ova (9.4, 10.5 and 12.0) and transferable embryos (5.3, 12.0 and 6.5) did not significantly differ among the control, T1 and T2 groups, respectively (p>0.05). However, mean concentrations of serum $P_4$ of the T1 (64.2 ng/ml) and T2 groups (55.7 ng/ml) were higher than that of control group (43.3 ng/ml, p<0.01), while serum cholesterol concentrations in the control (105.8 mg/dl) and T2 groups ($96.9\;{\pm}\;mg/dl$) were significantly lower than in the T1 group (121.1 mg/dl, p<0.05). Conclusively, whole crop barley silage can be fed a good substitute for hay forage for Hanwoo donors. Furthermore the ratios of whole crop barley silage 60% and rice straw 40% might be more worthful for embryo production.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1529-1536
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• 2006

We cloned a gene of ompH(D:4) from pigs infected with P. multocida D:4 in Korea [16]. The gene is composed of 1,026 nucleotides coding 342 amino acids (aa) with a signal peptide of 20 aa (GenBank accession number AY603962). In this study, we analyzed the ability of the ompH(D:4) to induce protective immunity against a wild-type challenge in mice. To determine appropriate epitope(s) of the gene, one full and three different types of truncated genes of the ompH(D:4) were constructed by PCR using pET32a or pRSET B as vectors. They were named ompH(D:4)-F (1,026 bp [1-1026] encoding 342 aa), ompH(D:4)-t1 (693 bp [55-747] encoding 231 aa), ompH(D:4)-t2 (561 bp [187-747] encoding 187 aa), and ompH(D:4)-t3 (540 bp [487-1026] encoding 180 aa), respectively. The genes were successfully expressed in Escherichia coli BL21(DE3). Their gene products, polypeptides, OmpH(D:4)-F, -t1, -t2, and -t3, were purified individually using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Their $M_rs$ were determined to be 54.6, 29, 24, and 23.2 kDa, respectively, using SDS-PAGE. Antisera against the four kinds of polypeptides were generated in mice for protective immunity analyses. Some $50{\mu}g$ of the four kinds of polypeptides were individually provided intraperitoneally with mice (n=20) as immunogens. The titer of post-immunized antiserum revealed that it grew remarkably compared with pre-antiserum. The lethal dose of the wild-type pathogen was determined at $10{\mu}l$ of live P. multocida D:4 through direct intraperitoneal (IP) injection, into post-immune mice (n=5, three times). Some thirty days later, the lethal dose ($10{\mu}l$) of live pathogen was challenged into the immunized mouse groups [OmpH(D:4)-F, -t1, -t2, and -t3; n=20 each, two times] as well as positive and negative control groups. As compared within samples, the OmpH(D:4)-F-immunized groups showed lower immune ability than the OmpH(D:4)-t1, -t2, and -t3. The results show that the truncated-OmpH(D:4)-t1, -t2, and -t3 can be used for an effective vaccine candidate against swine atrophic rhinitis caused by pathogenic P. multocida (D:4) isolated in Korea.

• Journal of Microbiology and Biotechnology
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• 제26권6호
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• pp.1115-1123
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• 2016

_L-Asparaginase (E.C. 3.5.1.1) is an enzyme involved in asparagine hydrolysis and has the potential to effect leukemic cells and various other cancer cells. We identified the _L-asparaginase gene (_L-ASPG86) in the genus Mesoflavibacter, which consists of a 1,035 bp open reading frame encoding 344 amino acids. Following phylogenetic analysis, the deduced amino acid sequence of _L-ASPG86 (_L-ASPG86) was grouped as a type I asparaginase with respective homologs in Escherichia coli and Yersinia pseudotuberculosis. The _L-ASPG86 gene was cloned into the pET-16b vector to express the respective protein in E. coli BL21 (DE3) cells. Recombinant _L-asparaginase (r-_L-ASPG86) showed optimum conditions at 37-40℃, pH 9. Moreover, r-_L-ASPG86 did not exhibit glutaminase activity. In the metal ions test, its enzymatic activity was highly improved upon addition of 5 mM manganese (3.97-fold) and magnesium (3.35-fold) compared with the untreated control. The specific activity of r-_L-ASPG86 was 687.1 units/mg under optimum conditions (37℃, pH 9, and 5 mM MnSO₄).

• Journal of Microbiology and Biotechnology
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• 제22권10호
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• pp.1330-1335
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• 2012

Foot and mouth disease (FMD) is one of the acute infectious diseases in hoofed and even-toed mammals, including pigs, and it occurs via acute infection by Aphthovirus. When FMD is suspected, animals around the location of origin are typically slaughtered and buried. Other methods such as rendering, composting, and incineration have not been verified in practice in Korea. After the FMD incident, the regular monitoring of the microbial community is required, as microorganisms greatly modify the characteristics of the ecosystem in which they live. This is the result of their metabolic activities causing chemical changes to take place in the surrounding environment. In this study, we investigated changes in the microbial community during a 24 week period with DNA extracts from leachate, formed by the decomposition of buried pigs at a laboratory test site, using denaturing gradient gel electrophoresis (DGGE) with a genomic DNA. Our results revealed that Bacteroides coprosuis, which is common in pig excreta, and Sporanaerobacter acetigenes, which is a sulfur-reduced microbe, were continuously observed. During the early stages (0~2 weeks) of tissue decomposition, Clostridium cochlearium, Fusobacterium ulcerans, and Fusobacterium sp., which are involved in skin decomposition, were also observed. In addition, various microbes such as Turicibacter sanguinis, Clostridium haemolyticum, Bacteroides propionicifaciens, and Comamonas sp. were seen during the later stages (16~24 weeks). In particular, the number of existing microbial species gradually increased during the early stages, including the exponential phase, decreased during the middle stages, and then increased again during the later stages. Therefore, these results indicate that the decomposition of pigs continues for a long period of time and leachate is created continuously during this process. It is known that leachate can easily flow into the neighboring environment, so a long-term management plan is needed in burial locations for FMD-infected animals.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1665-1672
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• 2012

An 80% ethanol extract of Hizikia fusiforme was obtained and followed by successive fractionation using the organic solvents n-hexane, ethyl acetate, and n-butanol to identify the antioxidative substance. The aqueous part of the nbutanol fractionation step, showing high antioxidative activity, was subjected to reverse-phase liquid chromatography. As a result, a substance purified from a BB-2 fraction showed high antioxidative activity. The m/z 419 [M+H] molecular ion peak in the fraction was observed by the analysis of the ESI-LC/MS spectrum. By the analysis of 1H NMR (500 MHz, DMSO-$d_6$) and $^{13}C$ NMR (125 MHz, DMSO-$d_6$) spectra, a unique compound of the fraction was biochemically identified as a 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5HHMF). We also investigated the effect of 5HHMF on human gastric AGS carcinoma cells. Western blot analysis suggested that the flavone substantially increased the levels of the death receptor-associated apoptosis mediators Fas, Fas L, FADD, TRADD, and DR4 in a concentration-dependent manner. The levels of Fas, Fas L, TRADD, and DR4 in the cells treated with 5HHMF ($5{\mu}g/ml$) were approximately 26.4-, 12.8-, 6.7-, and 9.8-times higher than those of non-treated cells, respectively. Of note, the level of FADD protein in the cells exposed to 5HHMF ($1{\mu}g/ml$) increased approximately 9.6-times. In addition, the cleavage of caspase-3, -8, and -9 in cultured AGS cells treated with 5HHMF was significantly confirmed. Therefore, our results suggest that 5HHMF from H. fusiforme is involved in the induction of death receptor-associated apoptosis mediators in human gastric AGS carcinoma cells.

• Journal of Microbiology and Biotechnology
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• 제30권9호
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• pp.1343-1354
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• 2020

Atopic dermatitis (AD) is a skin disorder that causes chronic itch. We investigated the inhibitory effects of a mixture of prebiotic short-chain galacto-oligosaccharides and long-chain fructooligosaccharides (scGOS/lcFOS), inulin, or β-glucan on AD development in 1-chloro-2,4-dinitrobenzene (DNCB)-treated NC/Nga mice. Mice were randomly assigned to six groups: untreated mice, AD control, positive control (DNCB-treated NC/Nga mice fed a dietary supplement of Zyrtec), and DNCB-treated NC/Nga mice fed a dietary supplement of prebiotics such as scGOS/lcFOS (T1), inulin (T2), or β-glucan (T3). The prebiotic treatment groups (T1, T2, and T3) showed suppression of AD symptoms, Th2 cell differentiation, and AD-like skin lesions induced by DNCB. In addition, prebiotic treatment also reduced the number of microorganisms such as Firmicutes, which is associated with AD symptoms, and increased the levels of Bacteroidetes and Ruminococcaceae, which are associated with alleviation of AD symptoms. Our findings demonstrate the inhibitory effects of prebiotics on AD development by improving the Th1/Th2 cytokine balance and beneficial symbiotic microorganisms in in vitro and in vivo models.

• Journal of Animal Science and Technology
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• 제60권1호
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• pp.29.1-29.6
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• 2018

Background: Dietary supplementation of zinc oxide (ZnO) to 2000 to 4000 mg/kg is known to be effective for the prevention and treatment of post-weaning diarrhea in the pig. Such a 'pharmacological' supplementation, however, can potentially result in environmental pollution of the heavy metal, because dietary ZnO is mostly excreted unabsorbed. Two experiments (Exp.) were performed in the present study to determine the effects of a lipid-coated ZnO supplement Shield Zn (SZ) compared with those of ZnO. Methods: In Exp. 1, a total of 240 21-day-old weanling pigs were fed a diet supplemented with 100 mg Zn/kg as ZnO (ZnO-100), ZnO-2500, SZ-100, or SZ-200 in 24 pens for 14 days on a farm with its post-weaning pigs exhibiting a low incidence of diarrhea. Exp. 2 was performed using 192 24-day-old piglets as in Exp. 1 on a different farm, which exhibited a high incidence of diarrhea. Results: In Exp. 1, fecal consistency (diarrhea) score (FCS) was less for the ZnO-2500 and SZ-200 groups than for the SZ-100 group (P < 0.05), with no difference between the SZ-100 and ZnO-100 groups. Both average daily gain (ADG) and gain:feed ratio were less for the SZ-200 group than for the ZnO-2500 group, with no difference between the ZnO-100 group and SZ-100 or SZ-200 group. The villus height (VH), crypt depth (CD), and VH:CD ratio of the intestinal mucosa were not influenced by the treatment. In Exp. 2, FCS was lowest for the ZnO-2500 group, with no difference among the other groups. However, neither the ADG nor gain:feed ratio was influenced by the treatment. Conclusion: Results suggest that physiological SZ supplementation has less beneficial effects than pharmacological ZnO for the alleviation of diarrhea irrespective of its severity and for promoting growth without influencing their integrity of the intestinal mucosal structures with little advantage over physiological ZnO in weanling pigs with a small pen size.