• Korean Journal of Acupuncture
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• 제32권3호
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• pp.108-115
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• 2015

Objectives : Acupuncture is frequently used as an alternative therapy for Parkinson's disease(PD) in Korea. Using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced Parkinson's disease mouse model, the present study investigated a possible role of acupuncture stimulation at GB34 in suppressing dopaminergic neuronal death and regulating the phosphorylation of protein kinase B(Akt) in substantia nigra(SN) and striatum(ST). Methods : Eight-week-old male C57BL/6 mice were administered intraperitoneally with 30 mg/kg of MPTP at 24-h intervals for 5 days. Acupuncture stimulation at GB34 or SI3 was performed once a day for 12 days consecutively from the first MPTP injection. After the last acupuncture stimulation, pole test was performed to assess the effect of the acupuncture stimulations. Dopaminergic neuronal survival in the SN and the ST, dopamine transporter( DAT) and caspase-3 expression in the ST were evaluated by immunohistochemistry. The phosphorylations of Akt in the SN and the ST were measured by Western blotting. Results : MPTP administration caused behavioral impairment and dopaminergic neuronal death in the SN and the ST. It also decreased DAT expression and increased caspase-3 expression in the ST. Acupuncture stimulation at GB34 alleviated these MPTP-induced impairments. Moreover, MPTP suppressed Akt phosphorylation in the SN and the ST, whereas acupuncture stimulation at GB34 alleviated the phosphorylation in the SN. Conclusions : These results indicate that acupuncture stimulation at GB34 can inhibit MPTP-induced dopaminergic neuronal death and alleviate the Akt phosphorylation in the SN, suggesting a possible role for acupuncture in the treatment of PD.

• 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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• 제32권2호
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• pp.187-194
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• 2022

Two α-ʟ-arabinofuranosidases (BfdABF1 and BfdABF3) and a β-ᴅ-xylosidase (BfdXYL2) genes were cloned from Bifidobacterium dentium ATCC 27679, and functionally expressed in E. coli BL21(DE3). BfdABF1 showed the highest activity in 50 mM sodium acetate buffer at pH 5.0 and 25℃. This exo-enzyme could hydrolyze p-nitrophenyl arabinofuranoside, arabino-oligosaccharides (AOS), arabinoxylo-oligosaccharides (AXOS) such as 3²-α-ʟ-arabinofuranosyl-xylobiose (A³X), and 2³-α-ʟ-arabinofuranosyl-xylotriose (A²XX), whereas hardly hydrolyzed polymeric substrates such as debranched arabinan and arabinoxylans. BfdABF1 is a typical exo-ABF with the higher specific activity on the oligomeric substrates than the polymers. It prefers to α-(1,2)-ʟ-arabinofuranosidic linkages compared to α-(1,3)-linkages. Especially, BfdABF1 could slowly hydrolyze 2³,3³-di-α-ʟ-arabinofuranosyl-xylotriose (A²⁺³XX). Meanwhile, BfdABF3 showed the highest activity in sodium acetate at pH 6.0 and 50℃, and it has the exclusively high activities on AXOS such as A³X and A²XX. BfdABF3 mainly catalyzes the removal of ʟ-arabinose side chains from various AXOS. BfdXYL2 exhibited the highest activity in sodium citrate at pH 5.0 and 55℃, and it specifically hydrolyzed p-nitrophenyl xylopyranoside and xylo-oligosaccharides (XOS). Also, BfdXYL2 could slowly hydrolyze AOS and AXOS such as A³X. Based on the detailed hydrolytic modes of action of three exo-hydrolases (BfdABF1, BfdABF3, and BfdXYL2) from Bf. dentium, their probable roles in the hemiceullose-utilization system of Bf. dentium are proposed in the present study. These intracellular exo-hydrolases can synergistically produce ʟ-arabinose and ᴅ-xylose from various AOS, XOS, and AXOS.

• Journal of Microbiology and Biotechnology
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• 제32권4호
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• pp.405-418
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• 2022

Simotang oral liquid (SMT) is a traditional Chinese medicine (TCM) consisting of four natural plants and is used to alleviate gastrointestinal side effects after chemotherapy and functional dyspepsia (FD). However, the mechanism by which SMT helps cure these gastrointestinal diseases is still unknown. Here, we discovered that SMT could alleviate gastrointestinal side effects after chemotherapy by altering gut microbiota. C57BL/6J mice were treated with cisplatin (DDP) and SMT, and biological samples were collected. Pathological changes in the small intestine were observed, and the intestinal injury score was assessed. The expression levels of the inflammatory factors IL-1β and IL-6 and the adhesive factors Occludin and ZO-1 in mouse blood or small intestine tissue were also detected. Moreover, the gut microbiota was analyzed by high-throughput sequencing of 16S rRNA amplicons. SMT was found to effectively reduce gastrointestinal mucositis after DDP injection, which lowered inflammation and tightened the intestinal epithelial cells. Gut microbiota analysis showed that the abundance of the anti-inflammatory microbiota was downregulated and that the inflammatory microbiota was upregulated in DDP-treated mice. SMT upregulated anti-inflammatory and anticancer microbiota abundance, while the inflammatory microbiota was downregulated. An antibiotic cocktail (ABX) was also used to delete mice gut microbiota to test the importance of gut microbiota, and we found that SMT could not alleviate gastrointestinal mucositis after DDP injection, showing that gut microbiota might be an important mediator of SMT treatment. Our study provides evidence that SMT might moderate gastrointestinal mucositis after chemotherapy by altering gut microbiota.

• Nutrition Research and Practice
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• 제18권1호
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• pp.19-32
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• 2024

BACKGROUND/OBJECTIVES: Atherosclerosis is associated with increased inflammation in the visceral adipose tissue (VAT). Vitamin D has been reported to modulate the inflammatory responses of stromal vascular cells (SVCs) and adipocytes in adipose tissue, but the role of vitamin D in atherosclerosis biology is unclear. This study examined the effects of in vitro 1,25-dihydroxyvitamin D₃ (1,25[OH]₂D₃) treatment on the inflammatory responses of SVCs and adipocytes from atherosclerotic mice. MATERIALS/METHODS: C57BL/6J (B6) mice were divided randomly into 2 groups and fed a 10% kcal fat control diet (control group, CON) or 41% kcal fat, 0.21% cholesterol (high fat + cholesterol, HFC) diet (obese group, OB), and B6.129S7-Ldlr^tm1Her/J (Ldlr^-/-) mice were fed a HFC diet (obese with atherosclerosis group, OBA) for 16 weeks. SVCs and adipocytes isolated from VAT were pre-incubated with 1,25(OH)₂D₃ for 24 h and stimulated with lipopolysaccarides for the next 24 h. Proinflammatory cytokine production by adipocytes and SVCs, the immune cell population in SVCs, and the expression of the genes involved in the inflammatory signaling pathway in SVCs were determined. RESULTS: The numbers of total macrophages and SVCs per mouse were higher in OB and OBA groups than the CON group. The in vitro 1,25(OH)₂D₃ treatment significantly reduced macrophages/SVCs (%) in the OBA group. Consistent with this change, the production of interleukin-6 and monocyte chemoattractant protein 1 (MCP-1) by SVCs from the OBA group was decreased by 1,25(OH)₂D₃ treatment. The 1,25(OH)₂D₃ treatment significantly reduced the toll-like receptor 4 and dual-specificity protein phosphatase 1 (also known as mitogen-activated protein kinase phosphatase 1) mRNA levels in SVCs and MCP-1 production by adipocytes from all 3 groups. CONCLUSIONS: These findings suggest that vitamin D can attribute to the inhibition of the inflammatory response in VAT from atherosclerotic mice by reducing proinflammatory cytokine production.

• 생명과학회지
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• 제32권2호
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• pp.135-141
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• 2022

Mouse thymosin β-4 (TB4) 유전자를 Escherichia coli에서 intein-tag 융합 단백질로 발현시키고 정제하였다. 재조합 TB4-intein 융합 단백질의 분자량을 10% glycine SDS-PAGE로 확인한 결과, 세포내 soluble 분획에서 60 kDa의 단백질을 얻을 수 있었고, 유도발현 최적 조건은 0.1 mM IPTG, 25℃에서 3시간 동안 유도 발현할 때가 최적임을 확인하였다. TB4 만을 얻기 위해서 유도발현된 TB4-intein을 DTT를 이용해 self-cleavage를 일으킨 다음, chitin bead를 이용한 친화성 크로마토그라피로 정제 후 분자량을 확인 한 결과, 5 kDa으로 확인되었으며, 순도는 95%이상 이었다. 정제한 recombinant TB4가 생물학적 기능을 보유하고 있는지 확인을 하기 위하여, TB4를 농도 별(1~1,000 ng/ml)로 하여 HT1080 cell을 이용한 cell migration을 측정한 결과, 모든 농도에서 recombinant TB4가 화학합성한 TB4 보다 약 20% 이상 높은 활성을 보였으며, recombinant TB4 1 ng/ml의 농도에서 cell migration 활성이 가장 높게 나타났다. Recombinant TB4를 마우스 상처 부위에 5일 동안 매일 처리한 결과(최종 처리 농도 0.5 mg/ml), 화학합성 TB4 보다 recombinant TB4의 상처치유 활성이 약 35% 더 높음을 알 수 있었다. 이상의 결과는 recombinant TB4가 화학합성 TB4보다 cell migration과 상처치유에 훨씬 높은 활성을 나타냄을 보여주고 있다.

• 현장농수산연구지
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• 제26권3호
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• pp.32-44
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• 2024

지리산에 자생하는 국화수리취(S. palmatopinnatifidus(Makino) Kitam.)), 생강나무(L. obtusiloba BL.), 석창포(A. gramineus Sol.), 쓴풀(S. japonica (Schult.) Griseb.), 쥐오줌풀(V. fauriei Briq.) 등 5종을 2022년 4월부터 10월까지 자생지에서 채집하였다. 채집한 식물은 건조 후 전초는 17종의 폴리페놀성분을 분석하고, 잎과 뿌리는 부위별에 따른 DPPH, ABTS의 항산화 활성을 측정하였으며, total polyphenol, total flavonoid의 함량을 측정하였다. 이 결과를 바탕으로 지리산에 자생하는 5종의 식물중항산화 효능이 높은 것을 대상으로 화장품 원료 항산화 화장품 원료 개발을 위한 기초자료로 활용되었으면 한다. 1. 지리산에 자생하는 5종의 식물에 함유된 폴리페놀성분은 catechin, quercetin을 포함한 12종이었다. 이중 석창포의 폴리페놀 총 함량은 39.5ppm으로 나타났고, 가장 낮은 폴리페놀 함량을 보인 품종은 쓴풀로 5.9ppm이었다. 2. DPPH radical 소거활성을 EC₅₀ 값으로 확인한 결과, DPPH radical 소거활성이 높은 것은 생강나무 (L. obtusiloba) 잎으로 35.31㎍/mL이었고, 가장 낮은 소거활성을 보인 것은 쥐오줌풀로 잎은 262.69 ㎍/mL, 뿌리는 278.99㎍/mL이었다. 3. ABTS radical 소거활성을 EC₅₀ 값으로 확인한 결과, 생강나무 뿌리는 55.79㎍/mL, 잎은 58.85㎍/mL으로 높게 나타났다. 가장 낮은 소거활성을 보인 것은 쓴풀 잎으로 576.5㎍/mL이었다. 4. 총폴리페놀의 함량이 가장 많은 품종은 생강나무 잎으로 158.72mg/g이었고, 함량이 가장 낮은 것은 쥐오줌풀 잎으로 0.13mg/g이었으며, 이는 품종에 따라 총폴리페놀의 함량은 큰 차이가 있음을 알 수 있었다. 5. 총플라보노이드의 함량이 가장 많은 품종은 생강나무로 잎에 163.75mg GAE/g이었고, 다른 품종들은 17.67~41.38mg GAE/g으로 낮은 함유량을 보였다.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.25-32
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• 1996

본 연구는 Polynucleotide-specific 형광물질을 이용한 Differential labelling 기법으로 체외수정 후 배양 4일째 생산된 B6CBA Fl 생쥐 배반포의 Total, ICM, Trophectoderm의 세포수를 조사함으로서 생쥐의 착상전 후기 배발달에 대한 기초 자료를 얻고자 실시하였다. 공시 배반포는 과배란처리에 의해 얻어진 난자를 $1{\times}10^6cells/ml$의 정자로 수정시키고, 95시간동안 M16배양액과 $37^{\circ}C$, 5% $CO_2$배양기 내에서 배양하여 배반포강의 확대와 투명대 두께의 감소를 기준으로 early, middle, expanded와 hatching으로 구분하였다. 본 연구에서 얻어진 결과는 다음과 같다. 1) 체외수정 후 95시간째 배반포 발달율은 86.7%였으며, early, middle, expanded와 hatching으로 16.3%, 18.9%, 10.5%, 40.9% 였다. 2) Bisbenzirnide를 이용한 배반포의 총 세포수는 early, middle, expanded, hatching 각각 $35.6{\pm}10.4$, $49.4{\pm}8.6$, $60.8{\pm}10.7$ 과 $62.7{\pm}13.9$를 얻었다. 3) Polynucleotide-specific형광물질을 이용한 Differential labelling으로 배반포 ICM과 Trophectoderm의 세포수를 early, middle, expanded, hatching으로 나누어 조사한 결과, ICM세포수는 각각 $9.6{\pm}3.0$, $13.6{\pm}3.9$, $16.0{\pm}3.3$, $19.5{\pm}4.6$개 이었고, Trophectoderm세포수는 $30.6{\pm}5.1$, $39.9{\pm}5.8$, $42.2{\pm}8.1$, $43.7{\pm}11.1$ 개로 나타나 ICM과 Trophectoderm 모두 동일하게 발달의 진행정도에 따라 세포수의 증가양상을 나타내었다. 또한, Bisbenzimide와 Differential labelling에서 얻어진 총세포수의 비교에서도 동일하게 발달의 진행정도에 따라 세포수의 증가를 나타내었으며 그와 동시에 세포수도 거의 유사하였다. 이러한 결과로 미루어 볼때, Differential labelling을 이용한 빠르고도 간편한 세포수 계산법은 착상전 후기 배발달을 고찰하는데 유용하며, 배양조건에 따른 Embryo의 Quality를 반영하는 Indicator로서 이용될 수 있다는 것을 시사한다.

• 한국식품저장유통학회지
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• 제11권3호
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• pp.373-382
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• 2004

재래적인 방법으로 제조된 죽력의 생리활성 효능을 구명하기 위하여 in vitro에서 항산화활성 및 HMG-CoA reductase 저해활성과 in vivo에서 고콜레스테롤혈증 개선효능을 실험하여 다음과 같은 결과를 얻었다 1. In vitro에서, Rancimat로 측정한 항산화활성은 죽력 1.25희석액과 원액은 대조구보다 높았고, HMG-Co A reductase저해활성은 죽력 원액이 57.9$\%$이었고 1.25희석액은 36.0$\%$이 었다. 2. In vivo에서, 6주에서 죽력 투여로 체중증가율은 대조군에 비하여 유의성있게 둔화되었고, 식이효율은 감소되었으나 유의적인 차이는 아니었으며, 간장/체중 비율은 실험군간에 유의성있는 변화를 나타내지 않았다. 죽력 저용량투여군의 총콜레스테롤량은 대조군에 비하여유의적인 감소를 나타내지 않았으나 고용량투여로 대조군보다 약 17$\%$정도 감소되었고, 중성지질량은 죽력 투여로대조군에 비하여 각각 26$\%$, 39$\%$의 유의적인 감소를 나타내었는데, 특히 고용량 투여군은 정상군보다 낮았으며, 인지질량은죽력투여로 대조군에 비하여 증가는 되었으나 유의적인 차이는 아니 었다. LDL-콜레스테를 농도는 고콜레스테롤식이 급여로 정상군보다 약 41$\%$ 정도 증가되었으나 죽력투여로 대조군보다 각각 12$\%$, 20$\%$씩 유의한 감소를 나타내었는데, 특히 고용량 투여군은 정상군의 농도에 근접하였으며, 죽력 투여로 HDL-콜레스테롤농도는 대조군에 비하여 상승되었으나 유의적인 효과는 아니었고, 죽력투여로 저하된 HDL-콜레스테롤/총콜레스테롤비는 대조군보다 약 48$\%$가 높아졌으며 동맥경화지수는 약 26$\%$가 낮아졌으나 유의성있는 변화를 나타내지는않았다. 고콜레스테롤식이 급여로 증가된 유리콜레스테를 농도는 고용량투여로 26$\%$가 감소되어 유의성있는 변화를 나타냈으며, 고콜레스테롤식이 급여로 콜레스테릴 에스테르 농도는정상군에 비하여 약 96$\%$이상의 증가를 나타냈으나 죽력 고용량 투여로 대조군에 비하여 32$\%$정도 감소되었다. 고콜레스테롤식이 급여로 간 중 총콜레스테롤량은 정상군에 비하여 유의하게 증가되었고, 죽력 저용량 투여는 증가된 총 콜레스테롤량을 감소시키지 못했으나 고용량 투여로 대조군보다 유의성있게 저하시켰으며, 중성지방량에는 변화가 없었다. 고콜레스테롤식이 급여로 상승된 ALT활성은 죽력투여로대조군에 비하여 유의하게 저하되었으나 ALT 및 ALP활성은 차이는 보이지 않았다. 이상의 실험결과에서 죽력이 in vitro에서 높은 항산화활성과 HMG-Co A reductase 활성을 유의하게 저해시켰고, in vivo에서 고콜레스테롤식이를 급여한 군보다 총콜레스테롤,LDL-콜레스테를 및 중성지질농도 등은 감소시켰으며, HDL-콜레스테롤, 인지질 농도 등은 증가시킴으로써지방간 및 동맥경화의 예방과 치료에 효과적일 것으로 추정되었다.

• Journal of Nutrition and Health
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• 제6권3호
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• pp.47-63
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• 1973

This study concerns the nutritional status of food in a lunch-box and the school lunch program provided for Dong Duck Elementary School Pupils in Daegu. Two hundred andsix pupils were randomly selected as a sample. The nutrients contained in a lunch-box and school lunch menu were analyzed according to the Food Composition Table. Nutrients and cost of the Dong Duck Elementary School Lunch were compared to the average nutrient contents of food in a lunch-box of o pupil. In addition, comparisons between the Recommended Daily Allowances of nutrients for Korean children aged from 9 to 11 (4th, 5th, and 6th grade) and that of the contained nutrients in the average lunch-box were conducted. The average nutrients contained in the lunch-box compared with the Recommended Daily Allowances are: Calorie 77.41% (542.60ca1), Protein 73.60% (16.93gr), Fat 33.95% (3.95gr), Calcium 27.85% (111.39mg), Ferret 80.30% (2.65mg), Vitamin A 10.09% (100.93 I.U.), Vitamin Bl 70.27% (0.27mg), Vitamin B2 88.37% (0.38mg), Vitamin C 15.40% (3.45mg). All of the nutrients in the lunch-box fall far below the Recommended Daily Allowances for the 9-11 age group. The average nutrients of staple food in the lunch-box compared with the side-dishes were found to contain Calorie 90.80%, Protein 50.32%, Fat 58.27%, Calcium 37.07 %, Ferret 53.96%, Vitamin A 0.0028%, Vitamin B1 78.11%, Vitamin B2 21.09%. The nutrients of food contained in the Dong Duck Elementary School Lunch Program compared with the Recommended Daily Allowances were: 1. The Protein, Fat, Ferret, Vitamin B1 and Vitamin B2 content was generally satisfactory. 2. Calcuim, Vitamin A and Vitamin C were lower than the Recommended Daily Allowances. 3. The Calorie content was generally lower for the 9-11 age group. The per pupil cost of the lunch-box was 41.87 won which includes the cost of staple food 16.92 won and side-dishes 24.95 won, whereas the school lune Provided by Dong Duck Elementary School Costs 35 won per pupil. Questionnaires were sent to four hundred parents (through their children) to find out their attitude toward the school lunch program. 390(97.50%) of them were returned and 7 of them dia not answer at all. Therefore, these were eliminated from the samlpe. Among 185 parents whose children were eating the school lunch; 172 parents (92.97% ) would pay 40 won for the proposed new school lunch program whereas only 13 parents (7.03%) did not want it. Among 198 parents whose children were not having the school lunch, 58 parents (29.29%) would agree to have the lunch for their children at the cost of 35 won with the proposed new school lunch program equivalent to 40 won. 126 parents (63.63%) could not agree to have their children being the school lunch because of their financial difficulty (74 parents 58.72%). The rest of parents (52 parents 41.28%) raised other reasons which were minor problems. Among the tatal of 383 parents, 309 parents (80.68%) would agree to gave school lunch for their children if the cost of the proposed new school lunch program would be 35 won to 40 won and if minor problems concerning the school lunch program are solved. Sixty parents (15.66% ) raised questions about the school lunch program concerning sanitary problems, taste of bread, and the quantity of lunch. Among 383 parents, 358 parents answered and 25 parents did not. When school officer asked if the parents would help during the school lunch hour, 101 parents indicated they could not help at all, but 177 parents, replied they could help. Among 177 parents, 40 parents of them could share their time from one or two hours everyday except Saturday and 107 parents could serve 1 to 3 days per week. Through this study, it is evident that further investigation should be done of the school lunch program which can be provided 35 won with the proposed new school lunch program. The parents participation in the school lunch program would be improved the understanding between school and parents as well as insure better nutrition and sanitation for the children.