• Journal of Veterinary Science
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• 제21권3호
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• pp.36.1-36.9
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• 2020

Background: Pseudorabies, also known as Aujeszky's disease, is caused by the pseudorabies virus (PRV) and has been recognized as a critical disease affecting the pig industry and a wide range of animals around the world, resulting in great economic losses each year. Shandong province, one of the most vital food animal-breeding regions in China, has a very dense pig population, within which pseudorabies infections were detected in recent years. The data, however, on PRV epidemiology and coinfection rates of PRV with other major swine diseases is sparse. Objectives: This study aimed to investigate the PRV epidemiology in Shandong and analyze the current control measures. Methods: In this study, a total number of 16,457 serum samples and 1,638 tissue samples, which were collected from 362 intensive pig farms (≥ 300 sows/farm) covered all cities in Shandong, were tested by performing enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Results: Overall, 52.7% and 91.5% of the serum samples were positive for PRV-gE and -gB, respectively, based on ELISA results. In addition, 15.7% of the tissue samples were PCR positive for PRV. The coinfection rates of PRV with porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus, and classical swine fever virus were measured; coinfection with PCV2 was 35.0%, higher than those of the other two viruses. Macroscopic and microscopic lesions were observed in various tissues during histopathological examination. Conclusions: The results demonstrate the PRV prevalence and its coinfection rates in Shandong province and indicate that pseudorabies is endemic in pig farms in this region. This study provides epidemiological data that can be useful in the prevention and control of pseudorabies in Shandong, China.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.52-52
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• 2001

Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic reaction follows a ping-pong mechanism in which the enzyme is transiently phosphorylated on a histidine residue conserved in all nucleoside diphosphate kinases. Beside their role in nucleotide synthesis, these enzymes present additional functions, possibly independent of catalysis, in processes such as differentiation, cell growth, tumor progression, metastasis and development. To clone murine nm23-M5, several expressed sequence tags (ESTs) of the GenBank data base, selected according to their homology to nm23-H5 cDNA, reconstituted a complete open reading frame (GenBank AF222750). To test whether murine NDPKs (1, 2, 3, 4, 5, and 6) can inhibit Bax-mediated toxicity in yeast, co-transformation was performed respectively. The yeast S.cerevisiae was transformed with a copy expression plasmid containing the histidine selection marker and expressing murine Bax under the control of a galactose-inducible promoter. Several clones were selected and found to be growth inhibited when Bax expression was induced with galactose. A representative clone was transformed again with a copy expression plasmid containing the tryptophane selection marker and expressing either murine Bcl-xL or NDPK under the control of a galactose-inducible promoter. Several subclones of the double-transformants were selected and characterized. The ability of Bcl-xL and NDPKs to suppress Bax-mediated toxicity was determined by growing yeast cells overnight in galactose media and spot-testing on galactose plates starting with an equal number of yeast cells as determined by taking the OD$_{600}$. Ten-fold serial dilutions were used in the spot-test. Plates were grown at 3$0^{\circ}C$ for 2-3 days. All murine NDPKs suppressed Bax dependent apoptosis. Futher study will be peformed whether Bax-toxicity inhibition was caused by NDP kinase activity or additional function.n.

• Journal of Ginseng Research
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• 제47권1호
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• pp.97-105
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• 2023

Background: Hyperactivated airway mucosa cells overproduce mucin and cause severe breathing complications. Here, we aimed to identify the effects of saponins derived from Panax ginseng on inflammation and mucin overproduction. Methods: NCI-H292 cells were pre-incubated with 16 saponins derived from P. ginseng, and mucin overproduction was induced by treatment with phorbol 12-myristate 13-acetate (PMA). Mucin protein MUC5AC was quantified by enzyme-linked immunosorbent assay, and mRNA levels were analyzed using quantitative polymerase chain reaction (qPCR). Moreover, we performed a transcriptome analysis of PMA-treated NCI-H292 cells in the absence or presence of Rg5, and differential gene expression was confirmed using qPCR. Phosphorylation levels of signaling molecules, and the abundance of lipid droplets, were measured by western blotting, flow cytometry, and confocal microscopy. Results: Ginsenoside Rg5 effectively reduced MUC5AC secretion and decreased MUC5AC mRNA levels. A systematic functional network analysis revealed that Rg5 upregulated cholesterol and glycerolipid metabolism, resulting in the production of lipid droplets to clear reactive oxygen species (ROS), and modulated the mitogen-activated protein kinase and nuclear factor (NF)-kB signaling pathways to regulate inflammatory responses. Rg5 induced the accumulation of lipid droplets and decreased cellular ROS levels, and N-acetyl-ⳑ-cysteine, a ROS inhibitor, reduced MUC5AC secretion via Rg5. Furthermore, Rg5 hampered the phosphorylation of extracellular signal-regulated kinase and p38 proteins, affecting the NF-kB signaling pathway and pro-inflammatory responses. Conclusion: Rg5 alleviated inflammatory responses by reducing mucin secretion and promoting lipid droplet-mediated ROS clearance. Therefore, Rg5 may have potential as a therapeutic agent to alleviate respiratory disorders caused by hyperactivation of mucosa cells.

• Animal Bioscience
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• 제36권7호
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• pp.1022-1033
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• 2023

Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.

• Journal of Microbiology and Biotechnology
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• 제33권4호
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• pp.430-440
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• 2023

The type three secretion system (T3SS) is a major virulence system of Pseudomonas aeruginosa (P. aeruginosa). The effector protein Exotoxin S (ExoS) produced by P. aeruginosa is secreted into the host cells via the T3SS. For the purpose of an experiment on inhibitors with regard to ExoS secretion, we developed a sandwich-type enzyme-linked immunosorbent assay (ELISA) system. Quercetin was selected because it has a prominent ExoS inhibition effect and also is known to have anti-inflammatory and antioxidant effects on mammalian cells. In this study, we investigated the effects of quercetin on the expression and secretion of ExoS using ELISA and Western blot analysis methods. The results showed that the secretion of ExoS was significantly decreased by 10 and 20 µM of quercetin. Also, popB, popD, pscF, and pcrV which are composed of the T3SS needle, are reduced by quercetin at the mRNA level. We also confirmed the inhibitory effect of quercetin on cytokines (IL-6, IL-1β, and IL-18) in P. aeruginosa-infected H292 cells by real-time polymerase chain reaction (PCR) and ELISA. Collectively, quercetin inhibits the secretion of ExoS by reducing both ExoS production and the expression of the needle protein of T3SS. Furthermore, these results suggest that quercetin has the potential to be used as an anti-toxic treatment for the inflammatory disease caused by P. aeruginosa infection.

• 한국식생활문화학회지
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• 제23권4호
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• pp.507-513
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• 2008

The development of soy cutlets containing textured soy protein (TSP) as a meat analog was studied. In order to decrease the beany flavor and to increase the texture, TSP was treated with 0.3% Flavourzyme or 0.1% Protamex for 10 or 20 min, respectively. The degree of hydrolysis for TSP treated with Protamex was higher than that treated with Flavourzyme. Hydrolysis was observed to increase as the reaction time was increased for both Flavourzyme and Protamex. The water holding capacity of TSP treated with Protamex for 10 min was the highest, and that treated with Flavourzyme for 20 min was similar to that of Protamex treatment for 20 min. The oil binding capacity of TSP treated with Protamex for 20 min was the highest. The hardness of the soy cutlets using TSP treated with Flavourzyme for 10 min was higher than that treated for 20 min, while that of Protamex treated for 20 min was higher than that treated for 10 min. The cohesiveness of the soy cutlets using TSP treated with Flavourzyme or Protamex for 10 min was higher than those treated for 20 min. The chewiness of the soy cutlets treated with Flavourzyme for 10 min was higher than for those treated for 20 min, while those treated with Protamex for 20 min was higher than those treated for 10 min. The springiness of TSP treated with Flavourzyme for 20 min was higher than those treated for 10 min, and higher than those treated with Protamex for 10 or 20 min. For sensory evaluation, the beany flavor of the soy cutlets treated with Protamex for 20 min was the weakest. The flavor and chewiness of both a pork cutlet and a soy cutlet treated with Protamex for 20 min were the best. In the overall quality, soy cutlets treated with Protamex for 20 min was the most desirable. In conclusion, soy cutlets treated with 0.1% Protamex for 20 min could be a reasonable substitute of pork cutlets.

• Journal of Veterinary Science
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• 제24권4호
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• pp.52.1-52.13
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• 2023

Background: Mesenchymal stem cells (MSCs) have been investigated as therapeutic agents for inflammatory bowel disease (IBD). Stimulation of MSCs with pro-inflammatory cytokines is an approach to enhance their immunomodulatory effects. However, further investigation is required to support their application in immune-mediated disorders and companion animals. Objectives: This study aimed to assess the therapeutic effect of tumor necrosis factor (TNF)-α-stimulated feline adipose tissue-derived MSCs (fAT-MSCs) in a dextran sulfate sodium (DSS)-induced colitis mouse model. Methods: Colitis mice was made by drinking water with 3% DSS and fAT-MSCs were injected intraperitoneally. Colons were collected on day 10. The severity of the disease was evaluated and compared. Raw 264.7 cells were cultured with the conditioned medium to determine the mechanism, using quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Results: TNF-α-stimulated fAT-MSCs more improved severity of DSS-induced colitis in disease activity, colon length, histologic score, and inflammatory cytokine. In sectionized colon tissues, the group comprising TNF-α-stimulated fAT-MSCs had higher proportion of CD11b⁺CD206⁺ macrophages than in the other groups. In vitro, TNF-α-stimulation increased cyclooxygenase-2 (COX-2) expression and prostaglandin E₂ (PGE₂) secretion from fAT-MSCs. The conditioned medium from TNF-α-stimulated fAT-MSCs enhanced the expression of interleukin-10 and arginase-1 in LPS-activated Raw 264.7 cells. Conclusions: These results represent that TNF-α-stimulated fat-mscs ameliorate the inflamed colon more effectively. Furthermore, we demonstrated that the effectiveness was interlinked with the COX-2/PGE₂ pathway.

• Animal Bioscience
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• 제37권3호
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• pp.509-521
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• 2024

Objective: It was shown that microRNAs (miRNAs) play an important role in milk protein synthesis. However, the post-transcriptional regulation of casein expression by exogenous miRNA (xeno-miRNAs) in ruminants remains unclear. This study explores the regulatory roles of alfalfa xeno-miR162 on casein synthesis in bovine mammary epithelial cells (bMECs). Methods: The effects of alfalfa xenomiR-162 and G protein subunit gamma 11 (GNG11) on proliferation and milk protein metabolism of bMECs were detected by 5-Ethynyl-2'-Deoxyuridine (EdU) staining, flow cytometry, cell counting kit-8 (CCK-8), enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot. Dual-luciferase reporter assay was used to verify the targeting relationship between GNG11 and xenomiR-162. Results: Results showed that over-expression of xenomiR-162 inhibited cell proliferation but promoted apoptosis, which also up-regulated the expression of several casein coding genes, including CSN1S1, CSN1S2, and CSN3, while decreasing the expression of CSN2. Furthermore, the targeting relationship between GNG11 and xenomiR-162 was determined, and it was confirmed that GNG11 silencing also inhibited cell proliferation but promoted apoptosis and reduced the expression of casein coding genes and genes related to the mammalian target of rapamycin (mTOR) pathway. Conclusion: Alfalfa xenomiR-162 appears to regulate bMECs proliferation and milk protein synthesis via GNG11 in the mTOR pathway, suggesting that this xeno-miRNA could be harnessed to modulate CSN3 expression in dairy cows, and increase κ-casein contents in milk.

• Parasites, Hosts and Diseases
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• 제24권1호
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• pp.25-41
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• 1986

뇌낭미충증환자 혈청 및 뇌척수액의 특이 IgG 항체가를 면역효소측정 법으로 측정하였을 때에 이 혈청학적 진단법이 환자진단에 얼마나 유용한지를 평가하였다. 1984년 1월부터 1986년 1월까지 주로 신경학적 중상을 나타낸 환자 355명에서 혈청 및 뇌척수액을 검사하였다. 면역효소측정법에 사용한 항원은 돼지에 자연감염된 유구낭미충의 낭액이며 단백질 농도 $2.5{\mu}g/ml$로 희석하여 사용하였고, 혈청은 1 : 100으로 희석하여, 뇌척수액은 희석하지 않고 반응시켰고 Perozidase-Conjugated Antihuman IgG goat serum(Cappel 회사제품)을 1 : 5,000으로 사용하여 혈청 및 뇌척수액내 유구낭미충 특이 IgG 항체가를 흡광도로 표시하였다. 흡광도 0.18 또는 그 이상을 양성으로 판정하였다. 그 결과를 요약하면 다음과 같다. 1. 대상자 355명중 신경외과 수술 및 병리학적 소견으로 확진된 뇌낭미충증 환자 26명, 피하결절 생검에서 낭미충증을 진단하였고 뇌전산화 단충촬영으로 확진한 환자 24명, 뇌전산화 단층환영으로 확진한 21명등 71명에서 면역효소측정법에 의한 특이 IgG항체가 양성자는 64명으로 민감도는 90.1%이었다. 그 중 혈청의 검사에 의한 민감도는 77.5%, 뇌척수액 검사에 의한 민감도는 83.1%로서 뇌척수액 검사가 더 민감한 소견이었다. 뇌낭미충증으로 확진된 환자중 위음성자는 특이 IgG 항체가가 대단히 낮은 예가 대부분이었다. 2. 대상자 355명중 뇌낭미충중 이외의 질환으로 확진된 환자는 52명으로서 그중 7례는 신경외과 수술 및 병리학적 소견에 근거하여 기타 질환으로 확진된 예이며 45례는 세균학적, 방사선학적 소견등을 근거로 기타 질환으로 확진된 예이었다. 이들 중 뇌낭미충 특이항체 검사에서 음성을 보인 예는 46례로서 이 검사의 특이도는 88.5%이었다. 혈청 및 뇌척수액검사에 의한 특이도는 카각 94.2%이었다. 위 양성반응을 보인 예 중에서 혈청 및 뇌척수액에서 모두 양성인 예는 없었다. 3. 혈청내 특이 IgG 항체 검사에 의한 기타 기생충감염자에서의 교차반응의 정도는 다음과 같았다. 무구조충증 18례중 2례, 스파르가눔증 20례중 2례, 폐흡충증 56례중 1례, 간흡충증 15례중 1례, 간질(간질)증 1례중 1례등이 교차반응을 나타내었다. 뇌척수액내 특이 IgG 함체 검사에 의한 교차반응을 뇌폐흡충증 환자 9례 중에는 없었으나 뇌스파르가눔중 환자 10례중 2례는 교차반응을 보였다. 뇌낭미충증으로 확진된 71례중 폐흡충항원에 대해 교차반응을 보인 예는 없었으나 스파르가눔 항원에 대해서는 혈청으로 검사했을 경우 6례, 뇌척수액의 경우 11례에서 교차반응을 나타내었다. 4. 뇌압상승이 있는 뇌낭미충증환자 예에서 뇌실조영술이나 뇌실복막강 단락술 도중 얻은 측뇌실 뇌척수액으로 낭미충 특이 IgG 항체가를 측정하면 측뇌실에 병변이 있지 않는 한 음성 또는 낮은 양성림위의 홉광도를 보이고 있었다. 5. 포도낭미충증 환자 4례중 혈청검사로는 4례중 3례가, 뇌척수액검사로는 검사한 3례 모두가 양성반응을 보였다. 이상의 결과는 혈청 및 뇌척수액내 특이 IgG 항체를 면역효소 측정 법으로 측정하는 혈청학적 진단법이 뇌낭미충증 환자의 감별진단에 매우 유용함을 보이고 있다.

• Korean Chemical Engineering Research
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• 제49권3호
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• pp.330-337
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• 2011

본 연구에서는 자기조립단분자막(SAM)의 연구에 유용한 일련의 $\omega$-mercapto alkyl amine 1 및 $\omega$-mercapto alkanoic acid 2를 만드는 제조공정을 서술하였다. 먼저 첫 번째 목표물질인 $\omega$-mercapto alkyl amine 1의 제조방법은 다음과 같다: 먼저 시중에서 시판되는 potassium phthalimide와 $\omega$-bromo alcohol을 $80{^{\circ}C}$, DMF 용매에서 치환반응으로 화합물 3을 합성하였다. 아민기와 알코올기를 포함하는 화합물 4을 합성하기 위하여 먼저 화합물 3를 hydrazine hydrate로 환류시킨 후, 이어서 c-HCl로 처리하여 부반응물 5를 수반하여 생성물 4를 76-98% 수율로 만들었다. $\omega$-aminoakanol 4의 hydroxyl기를 HBr로 브로민화반응을 하여 $\omega$-bromoamine 화합물 6을 34-97% 수율로 만들었다. 티올기를 도입하기 위하여 화합물 6을 95% 에탄올 속에서 thiourea와 치환 반응하여 $\omega$-aminoalkanthiuronium 7을 만든 다음, 이 화합물을 센 염기(KOH)와 센 산으로 처리하여 목표화합물, $\omega$-mercapto alkylamines 1을 총 5단계를 걸쳐서 제조하였다. 덧붙여 두 번째 목표물질인 $\omega$-mercapto alkanoic acid 2는 다음과 같이 2단계를 통하여 제조하였다: $\omega$-bromo alkanoic acid를 95% 에탄올 속에서 thiourea로 처리하여 화합물 7을 만든 다음, 센 염기(KOH)를 처리하여 thiuronium bromide 를 제거한 후, 다시 센 산(HCl) 수용액으로 처리하여 두 번째 목표화합물 $\omega$-mercapto alkanoic acid 2을 얻었다. 제조한 긴 사슬 알킬티올 1과 2 유도체들은 금속(Au, Pt, Ti)에 자기조립단분자 막을 형성함으로써 단백질, 효소 및 다양한 생체분자를 고정하는 연결자로 사용될 수 있으며, 그 밖에 금속의 표면개질을 이용하여 다양한 응용 연구를 위한 화학 도구로 사용될 수 있다.