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Iron-fortified recombinant Saccharomyces cerevisiae producing Sus scrofa ferritin heavy-chain recovers iron deficiency in mice

  • Lim, Hwan;Kim, Jong-Taek;Kim, Myoung-Dong;Rhee, Ki-Jong;Jung, Bae Dong
    • Korean Journal of Veterinary Research
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    • v.52 no.4
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    • pp.263-268
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    • 2012
  • In this study, we produced iron-fortified yeast (Saccharomyces cerevisiae) producing Sus scrofa ferritin heavy-chain to provide iron supplementation in anemic piglets. We determined whether iron-ferritin accumulated in recombinant yeasts could improve iron deficiency in mice. C57BL/6 male mice exposed to Fe-deficient diet for 2 weeks were given a single dose of ferrous ammonium sulfate (FAS), ferritin-producing recombinant yeast (APO), or APO reacted with iron ($Fe^{2+}$) (FER). The bioavailability of recombinant yeasts was examined by measuring body weight gain, hemoglobin concentration and hematocrit value 1 week later. In addition, ferritin protein levels were evaluated by western blot analysis and iron stores in tissues were measured by inductively coupled plasma spectrometer. We found that anemic mice treated with FER exhibited increased levels of ferritin heavy-chain in spleen and liver. Consistently, this treatment restored the iron concentration in these tissues. In addition, this treatment significantly increased hemoglobin value and the hematocrit ratio. Furthermore, FER treatment significantly enhanced body weight gain. These results suggest that the iron-fortified recombinant yeast strain is bioavailable.

Aronia melanocarpa reduced adiposity via enhanced lipolysis in high-fat diet-induced obese mice

  • Kim, Hyun Kyung;Jung, Jiyeon;Kang, Eun Young;Gang, Gyoungok;Kim, Wooki;Go, Gwang-woong
    • Korean Journal of Food Science and Technology
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    • v.52 no.3
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    • pp.255-262
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    • 2020
  • Obesity is a critical health issue in Korea, where half of all adults are overweight and a third obese. Aronia melanocarpa -rich in flavonoids and phenolics- with antioxidant and anti-inflammatory activities, could have anti-obesity activity and reduce body fat mass by upregulating lipolysis and β-oxidation in obese mice. Male C57BL/6J mice (n=12) were assigned into four groups: normal chow (18% kcal from fat); high-fat diet control (HFD, 45% kcal from fat); HFD+A. melanocarpa (200 mg/kg diet); HFD+Xenical (500 mg/kg diet, positive control). Antioxidant capacity of A. melanocarpa was established in vitro and in vivo. Weight loss was induced as decreased adiposity and lowered respiratory quotient at rest suggested oxidation of stored fat. Adiposity reduction, accompanied with elevated fat utilization, was owing to enhanced activity of hormone-sensitive lipase. Thus, A. melanocarpa lowered adiposity by enhancing lipolysis and utilization of fatty acids in visceral fat.

Clinical Review of the Effects of Hominis Placental Pharmacopuncture in the Treatment of Facial Spasm Patients

  • Jo, Na-Young;Kim, Jeong-Hyun;Roh, Jeong-Du
    • Journal of Pharmacopuncture
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    • v.16 no.3
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    • pp.52-57
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    • 2013
  • Objectives: The main purpose of this research is to investigate the effect of treatment with Hominis Placental pharmacopuncture (HPP) for 32 patients with hemifacial spasm. Methods: We treated facial spasm patients with acupuncture and HPP at Sabaek (ST2), Seung-eup (ST1), Gwallyeo (SI18), Chanjuk (BL2), Sajukgong (TE23), Hagwan (ST7), Hyeopgeo (ST6), Jichang (ST4), Wan-gol (SI4) and Yepung (TE17), and we investigated the effect by using Scott's scale. The data were analyzed by using the SPSS/10.0 for windows program with descriptive statistics, the paired t-test, and the Shapiro-Wilk normality test. Results: After treatment, the grade of the spasm's intensity based on Scott's description were decreased significantly. About 72% of the patients felt that the combination treatment had produced excellent results. Conclusion: These data suggested that HPP can be useful for treating facial spasm patients.

Studies on the Mechanism of the Ameliorative Activities on Hyperglycemia and Dyslipidemia of Red Ginseng Herbal Acupuncture in C57BL/KsJ db/db Mice (홍삼약침(藥鍼)이 제2형 당뇨병 동물모델의 항고혈당(抗高血糖)및 항고지질(抗高脂質) 기전(機轉)에 미치는 영향(影響))

  • Kim, Jong-Deok;Kim, Jong-In;Koh, Hyung-Kyun;Lee, Yun-Ho;Kang, Sung-Keel
    • Journal of Acupuncture Research
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    • v.25 no.2
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    • pp.11-26
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    • 2008
  • 목적 : 홍삼약침(藥鍼)이 고혈당 및 지질대사장애에 미치는 개선효과와 그 기전을 조사하고자 한다. 방법 : 홍삼약침(藥鍼)의 anti-diabetic 활성과 그 기전을 C57BL/KsJ db/db mice를 이용하여 관찰하였다. 실험 동물은 대조군(DC), 홍삼약침(藥鍼)군(RGL, RGH) 및 양성대조군(MET, GPZ, PIO)의 6군으로 나누었다. 홍삼약침(藥鍼)군은 $0.2m{\ell}$의 홍삼약침멸(藥鍼滅)을 각각 100mg/kg(RGL) 및 200mg/kg(RGH)씩 인체의 간유(肝兪)($BL_{18}$)에 상응하는 혈위에 1일 1회 10주간 좌우 혈을 번갈아가며 약침 시술하였다. 양성대조군은 metformin 300mg/kg(MET), glipizide 15mg/kg(GPZ) 및 pioglitazone 30mg/kg(PIO)을 각각 1일 1회 10주간 경구투여 하였다. 체중과 혈당은 매주 측정하였다. 실험 10주 후에는 혈액채취로 혈중 glucose, 당화혈색소(HbAlc), insulin, 중성지방(TG), adiponectin, leptin, non-esterified fatty acid(NEFA)를 측정 하였고, 간 조직을 채취하여 조직학적 검사 및 gene expression 분석을 시행하였다. 결과 : 홍삼약침(藥鍼)(RGL, RGH)은 10주 동안 C57BL/KsJ db/db mice의 체중을 증가시키는 부작용은 나타나지 않았다. 홍삼약침(藥鍼)군(RGL, RGH)의 사료섭취량은 대조군과 비슷하였으나 음수량은 증가하였다. 홍삼약침(藥鍼)(RGL, RGH)은 대조군에 비하여 각각 19.8% 및 18.3% 혈당을 낮추었고, 홍삼약침(藥鍼)(RGL)은 insulin resistance를 27.7% 감소시켰으며, 경구내당능 검사의 혈중 glucose에서는 대조군에 비해 홍삼약침(藥鍼)군(RGL, RGH)과 양성대조군(MET, GPZ, PIO)에서 각각 19.8%, 18.3%, 67.7%, 52.3% 및 56.9% 감소시켰다. 당화혈색소(HbAlc)는 홍삼약침(藥鍼)(RGL, RGH), MET, GPZ 및 PIO군에서 대조군에 비하여 각각 11.0%, 6.4%, 18.9%, 16.1% 및 27.9% 감소시켰으며, 혈중 glucose감소와 유사한 경향을 나타내었다. 홍삼약침(藥鍼)(RGL)은 대조군에 비해 TG와 NEFA를 각각 18.8% 및 16.8% 감소시켰고, adiponectin과 leptin을 각각 20.6% 및 12.1% 증가시켰다. 홍삼약침(藥鍼)(RGL, RGH)은 중성지방의 침착으로 인한 간의 질량비 증가를 억제하지 못하였으나, 지방구를 감소시겼음을 관찰할 수 있었다. Microarray 분석에서는 홍삼약침(藥鍼)(RGL, RGH)이 간에서 glycolysis, gluconeogenesis 및 fatty acid beta-oxidation과 관련된 유전자 발현에 영향을 미치는 것으로 나타나 양성대조군 metformin과 유사한 기전을 나타내었다. 요약 : 홍삼약침(藥鍼)은 T2DM동물모델(C57BL/KsJ db/db mice)에서 항당뇨 및 지질대사 개선활성이 있었다. 홍삼약침(藥鍼)은 C57BL/KsJ db/db mice의 간조직에서 lipogenesis억제 및 fatty acid beta-oxidation활성을 통해 혈당 이용을 높이고, insulin sensitivity를 향상시켰다. 또한 유전자 발현분석을 통해 그 기전이 metformin과 유사함을 확인할 수 있었으므로 향후 홍삼약침(藥鍼)의 새로운 약침 기술 개발 근거가 될 수 있을 것으로 사료된다.

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Xylanase Production by Mixed Culture Using Crude Hemicellulose from Rice Straw Black Liquor and Peat Moss as an Inert Support

  • Shata, Hoda Mohamed Abdel Halim;El-Deen, Azza Mohmed Noor;Nawwar, Galal Abdel Moen;Farid, Mohmed Abdel Fattah
    • Journal of Applied Biological Chemistry
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    • v.57 no.4
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    • pp.313-320
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    • 2014
  • Black liquor (BL) is a by-product of rice straw pulping process. It is a low costs raw material for production value-adding proteins and enzymes, which has been paid more and more attention to reduce its environmental pollution. Mixed cultures of micelial fungi, Trichoderma reesei Northern Regional Research Laboratory (NRRL)11236, Trichoderma reesei NRRL 6165 and Aspergillus niger strains NRC 5A, NRC 7A, and NRC 9A were evaluated for their ability to produce xylanase using crude hemicellulose (CHC) prepared from BL and peat moss as an inert support under solid state fermentation (SSF). The most potent strains, A. niger NRC 9A (818.26 U/g CHC) and T. reesei NRRL 6165 ($100.9{\pm}57.14$ U/g CHC), were used in a mixed culture to enhance xylanase production by co-culturing under SSF. In the mixed culture, xylanase production ($1070.52{\pm}12.57$ U/g CHC) was nearly1.3 and 10.6-fold increases over the activities attained in their monocultures, A. niger NRC 9A and T. reesei NRRL 6165, respectively. Optimization of the culture parameters of the mixed culture SSF process, concentration of ammonium sulfate and corn steep liquor, CHC/peat moss ratio, inoculum size and ratios of the two strains, initial pH value, initial moisture content and incubation time, exhibited a significant increase ($2414.98{\pm}84.02$ U/g CHC) in xylanase production than before optimization.

Cloning and Biochemical Characterization of Aspartate Aminotransferase from Xanthomonas oryzae pv. oryzae (Xanthomonas oryzae pv. oryzae로 부터 aspartate aminotransferase 유전자의 분리 및 생화학 특성)

  • Kang, Han-Chul;Yoon, Sang-Hong;Lee, Chang-Mook
    • Journal of Applied Biological Chemistry
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    • v.52 no.3
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    • pp.109-115
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    • 2009
  • The gene encoding a putative aspartate aminotransferase in Xanthomonas oryzae pv. oryzae (Xoo) was cloned using PCR technique. The gene was ligated with pET-21(a) vector containing His6 tag and expressed in E. coli BL21(DE3). Affinity purification of the recombinant aspartate aminotransferase with Ni-NTA resin resulted in one band by SDS-PAGE analysis. The purified enzyme showed a molecular weight of 43 kDa, as expected. The enzyme was the most active toward L-aspartate as an amino donor, indicating that the purified enzyme is one of aspartate aminotrans-ferases exist in Xoo. Optimal activity of the enzyme was observed at around pH 7.5 and stability was much higher at alkaline pH rather than acidic pH values. The enzyme was considerably activated by the presence of manganese ion, showing about 157% of control activity at 1.0 mM.

Anti-diabetic Mechannism Study of Korean Red Ginseng by Transcriptomics (전사체 프로파일을 이용한 고려 홍삼의 항당뇨 기전 연구)

  • Yuan, Hai-Dan;Shin, En-Jung;Chung, Sung-Hyun
    • YAKHAK HOEJI
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    • v.52 no.5
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    • pp.345-354
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    • 2008
  • This study was designed to investigate the anti-diabetic effect and mechanism of Korean red ginseng extract through transcriptomics in C57BL/KsJ db/db mice. The db/db mice were randomly divided into six groups: diabetic control group (DC), red ginseng extract low dose group (RGL, 100 mg/kg), red ginseng extract high dose group (RGH, 200 mg/kg), metformin group (MET, 300 mg/kg), glipizide group (GPZ, 15 mg/kg) and pioglitazone group (PIO, 30 mg/kg), and treated with drugs once per day for 10 weeks. At the end of treatment, we measured blood glucose, insulin, hemoglobin A1c (HbA1c), triglyceride (TG), adiponectin, leptin, non-esterified fatty acid (NEFA). RGL-treated group lowered the blood glucose and HbA1c levels by 19.6% and 11.4% compared to those in diabetic control group. In addition, plasma adiponectin and leptin levels in RGL-treated groups were increased by 20% and 12%, respectively, compared to those in diabetic control. Morphological analyses of liver, pancreas and epidydimal adipose tissue were done by hematoxylin-eosin staining, and pancreatic islet insulin and glucagon levels were detected by double-immunofluorescence staining. RGL-treated group revealed higher insulin contents and lower glucagon contents compared to diabetic control. To elucidate an action mechanism of Korean red ginseng, DNA microarray analyses were performed in liver and fat tissues, and western blot and RT-PCR were conducted in liver for validation. According to hierarchical clustering and principal component analysis of gene expression Korean red ginseng treated groups were close to metformin treated group. In summary, Korean red ginseng lowered the blood glucose level through protecting destruction of islet cells and shifting glucose metabolism from hepatic glucose production to glucose utilization and improving insulin sensitivity through enhancing plasma adiponectin and leptin levels.

Syphacia obvelata: A New Hope to Induction of Intestinal Immunological Tolerance in C57BL/6 Mice

  • Taghipour, Niloofar;Mosaffa, Nariman;Rostami-Nejad, Mohammad;Homayoni, Mohamad Mohsen;Mortaz, Esmaeil;Aghdaei, Hamid Asadzadeh;Zali, Mohammad Reza
    • Parasites, Hosts and Diseases
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    • v.55 no.4
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    • pp.439-444
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    • 2017
  • The ability of nematodes to manipulate the immune system of their host towards a Th2 and T regulatory responses has been proposed to suppress the inflammatory response. Clinical trials have proposed a useful effect of helminth infections on improvement of inflammatory disorders. In this study, we investigated the immunomodulatory effect of Syphacia obvelata infection to induce intestinal tolerance in C57BL/6 mice. Mice were infected through the cagemates with self-infected BALB/c mice. Four weeks post-infection, expression levels of $IFN-{\gamma}$, $TNF-{\alpha}$, IL-17, and IL-10 were assessed in the supernatant of mesenteric lymph node (MLN) culture. $Foxp3^+Treg$ were measured in MLN cells by flow cytometry. In the S. obvelata-infected group, the percentage of Tregs ($5.2{\pm}0.4$) was significantly higher than the control ($3.6{\pm}0.5$) (P<0.05). The levels of IL-10 ($55.3{\pm}2.2$ vs $35.2{\pm}3.2$), IL-17 ($52.9{\pm}3.8$ vs $41{\pm}1.8$), $IFN-{\gamma}$ ($44.8{\pm}4.8$ vs $22.3{\pm}2.3$) and $TNF-{\alpha}$ ($71.1{\pm}5.8$ vs $60.1{\pm}3.3$) were significantly increased in infected mice compared to the control group (P<0.05). The above results showed the potential effects of S. obvelata to induce intestinal tolerance. Therefore, it seems that S. obvelata may increase the immunological suppressive function in the intestinal tract.

Antitumor and Immunomodulatory Effects of Chungsangbohahwan on Murine Melanoma-induced Lung Metastasis (청상보하환(淸上補下丸)이 흑색종(黑色腫)의 폐전이(肺轉移) 억제(抑制) 및 면역조절작용(免疫調節作用)에 미치는 영향(影響))

  • Ha, Jee-Yong;You, Byeong-Gil
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.159-175
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    • 1998
  • In order to investigate the antitumor effect by Chungsangbohahwan after B-16 cells were transplanted in C57BL/6 mice, and the immune responses in mice induced by methotrexate, the extract of Chungsangbohahwan was orally administered to the mice for 21 days. Experimental studies were performed for measurance of metastasis, cell cytotoxicity in vitro, natural killer cell activity, productivity of interleukin-2. The results were summarized as follows: 1. Inhibition of metastasis in Chungsangbohahwan-treated group was higher than control group with significance on 7th day and 14th day. 2. On the MTT assay, cell viability was significantly inhibited by $5{\mu}g/well$, $2.5{\mu}g/well$, $1.25{\mu}g/well$, and $0.625{\mu}g/well$ of Chungsangbohahwan concentration inhibited cell viability significantly(P<0.05). $IC_{50}$ for cell viability was $2.17{\mu}g/well$. 3. Natural killer cell activity in Chungsangbohahwan-treated group was significantly increased on 100:1, 50:1 E/T(effect cell/target cell) ratio(P<0.05). 4. Production of interleukin-2 in Chungsangbohahwan-treated group was significantly increased(P<0.05).

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Lack of EGCG Effects on Radiation-Induced Apoptosis of Mice Splenocytes (마우스 비장세포에서 방사선유도성 아포토시스에 대한 EGCG 효과의 부족)

  • Jang, Seong-Soon;Lee, Heui-Kwan
    • Radiation Oncology Journal
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    • v.26 no.3
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    • pp.173-180
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    • 2008
  • Purpose: The modification of radiation-induced apoptosis by EGCG, known as antioxidants or oxidants, was studied in mice spleens irradiated with a lethal dose. Materials and Methods: Male C57BL/6 mice were divided into control, irradiation-only, and EGCG (100 mg/kg i.p. 1 h before irradiation) pretreatment groups. The mice were irradiated with a single whole-body dose of 7 Gy. The apoptosis in the spleens after irradiation of the lethal dose were analyzed by TUNEL assay. In addition, the expression levels of the Bax and Bcl-2 proteins were quantified using a Western blotting method. Results: The induction of apoptosis was detected in the splenic white pulp. The highest level of apoptosis was detected at 8 hours after irradiation. No significant difference was identified by the apoptotic index (53.9% vs. 52.1%, p=0.328) and relative Bax protein expression (0.86 vs. 0.81, p=0.335), between the irradiation-only and EGCG pretreatment group, respectively. However, a lower Bax/Bcl-2 ratio (1.64 vs. 0.97, p=0.037) and higher relative expression level of Bcl-2 protein (0.57 vs. 0.82, p=0.037) was measured in the EGCG pretreatment group. Conclusion: The EGCG pretreatment neither decreased the radiation-induced apoptosis in mice splenocytes, nor induced additional apoptosis.