• Title/Summary/Keyword: KIT-6

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Protease Activity of Lactic Acid Bacteria Isolated from Korean Traditional Fermented Food (전통 발효식품으로부터 Protease 활성을 보유한 유산균의 분리 및 동정)

  • Kook, Moo Chang;Cho, Seok Cheol;Park, Hoon;Kim, Seung Seop;Pyun, Yu Ryang;Choi, Woon Yong;Lee, Hyeon Yong
    • Food Engineering Progress
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    • v.15 no.2
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    • pp.182-187
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    • 2011
  • A proteolytic lactic acid bacterium was isolated from Korean traditional fermented foods. The isolate BV-26, which had a protease activity (24 U/mg-crude protein), was identified as Lactobacillus plantarum by the API 50CHL kit and 16S rDNA analysis (99.9% of homology), and named as L. plantarum BV-26. Cell growth and protease activity of L. plantarum BV-26 was determined in MRS broth using 5L jar fermentor at $30^{\circ}C$. The maximum growth of L. plantarum BV-26 was reached at 18 hr in MRS broth, while protease activity of BV-26 was detectable at 12 hr and the highest activity was obtained after 16 hr cultivation. Therefore, we expect that the proteolytic lactic acid bacteria, L. plantarum BV-26, may be used as a starter for the fermentation of animal feed. Especially, the fermentation of soybean meal with the strain can be applied for improving feed utilization.

Studies on the regulation of Hematopoietic enhancement of Brassica campestris var narinosa., Canavalia gladiata DC semen and their combinational prescription via Jak2/STAT5/GATA1 Pathway in Sca-1+ hematopoietic stem cells (Sca-1+골수조혈세포에서 JAK2/STAT5/GATA-1 신호전달 경로를 통한 다채, 도두 그리고 두 조합물에 의한 조혈증진 조절에 관한 연구)

  • Kim, Kunhoae;Kim, Seung-Hyung;Cho, In-Sik;Kim, Han-Young;Kim, Dong-Seon;Lee, Young-Cheol
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.7-16
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    • 2013
  • Objectives : Brassica campestris var narinosa (BCN), Canavalia gladiata DC semen (CGD) and their combinational prescription (BCN+CGD) have been use to demonstrate to regulate hematopoiesis. In the current study, we investigated whether Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription is related to hemato-potentiating function using Sca-$1^+$ hematopoietic stem cells (Sca-$1^+HSCs$) as a testing system. Methods : Sca-$1^+HSCs$ isolated from femur in C57bl/6 mice with leukopenia and thrombocytopenia induced by cyclophosphamide (CTX). Then, Real-time PCR was performed to measure the mRNA expression, ELISA and haematopoiesis-related gene (EPO, TPO, IL-3, SCF, c-kit, GM-CSF), the phosphorylation of JAK2, GATA-1 and STAT-5a/b were observed by western blot, and the numbers of $CD117^+/Sca-1^+$ cell and the number of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E), semisolid clonogenic assay was performed. Result : When Sca-$1^+HSCs$ were treated with Brassica campestris var narinosa, Canavalia gladiata DC semen and their combinational prescription with rIL-3/rSCF, the expression of haematopoiesis-related (EPO, TPO, IL-3, SCF, c-kit, and GM-CSF) were significantly increased at the levels of mRNA as well as production in Sca-$1^+HSCs$. Additionally, CGS enhanced phosphorylation of JAK2, GATA-1, and signal transducer and activator of transcription-5a/b (STAT-5a/b) in Sca-$1^+HSCs$. Furthermore, their combinational prescription (BCN+CGD) significantly enhanced the growth rate of granulocyte erythrocyte monocyte macrophage colony-forming units (CFU-GEMM) and erythroid burst forming units (BFU-E) in vitro. Conclusion : These result suggest that Brassica campestris var narinosa (BCN) and Canavalia gladiata DC have hematopoietic enhancement via hematopoietic cytokine-mediated JAK2/GATA-1/STAT-5a/b pathway, and their combinational prescription (BCN+CGD) has superior hematopoietic enhancement to those of individual extracts.

Seroepidemiological study of Toxoplasma gondii infection in the rural area Okcheon-gun, Korea

  • Lee, Young-Ha;Hoh, Hyung-Jun;Hwang, Ok-Sun;Lee, Sang-Keol;Shin, Dae-Whan
    • Parasites, Hosts and Diseases
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    • v.38 no.4
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    • pp.251-256
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    • 2000
  • There have been some reports about the prevalence of anti-Toxoplasma gondii antibody among Koreans, and most of all data were taken from patients visiting hospitals. However, the epidemiological data of the community-based study in Korea are rare. This study was performed to evaluate the seroprevalence of toxoplasmosis among the inhabitants of the rural area Okcheon-gun, Korea. A total of 1,109 serum samples (499 males, 610 females) were examined for the IgG antibodies by ELISA. To set up the cut-off point for ELISA, we used a commercial latex agglutination (LA) kit. The sensitivity and specificity of ELISA against LA test were 89.5%, and 98.6% respectively. Among 1,109 sera, 6.9% showed seropositivity by ELISA. The positive rates of males and females were 6.0% and 7.2%, respectively. However, there were no significant differences between sexes. Comparing the age groups, the highest seropositive rate showed in the seventies or higher, and their rates had a tendency to increase with age (0.05 < p < 0.3). These results revealed that the seroprevalence of toxoplasmosis in rural inhabitants is similar to previous reports in Korea; however we need further investigation to clarify the prevalence of toxoplasmosis in the general population.

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A study on Toxoplasmosis in dogs (견의 Toxoplasmosis에 관한 연구)

  • Lee, Cheong-San;Kwak, Hak-Koo;Park, Kyung-Jae;Jang, Hwan
    • Korean Journal of Veterinary Service
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    • v.21 no.1
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    • pp.29-39
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    • 1998
  • This study was carried out to investigate the prevalence of antibody against Toxoplasma gondii in the dog by Latex agglutination test and Indirect fluorescent antibody test. Two month-old dogs were infected intraperitoneally with T gondii to observe histopathological and immunohisto-chemical changes. Results obtained through this experiment were summarized as follows ; 1. Among the serum samples of 163 heads of the dog, 10 samples(6.1%) were positive. 2. In the sex, 6 heads (7.1%) out of 84 female dogs and 4 heads(5.1%) out of 79 male dogs were positive. However, there were no significant differences between the male and female. 3. Overall proportion of agreement between indirect fluorescent antibody and Latex agglutination test in 163 sera of dogs was 97.5%. 4. When 2 month-old dogs were infected intraperitoneally with T gondii, main clinical signs were intermittent fever, dyspnea, diarrhea. In general, the infected dogs recovered closely on the 11th day of post-inoculation. 5. At necropsy, petechial and ecchymotic hemorrages and swelling in small intestine, lung, spleen, liver and kidney were observed. 6. In histopathological observation, interstitial pneumonia, hyperemia and hemorrhages in lung were observed. Focal necrosis of hepatocytes, the neutrophil and basophil in the renal tubular epithelium were observed. 7. By immunohistochemical staining using Vectorstain ABC kit, the positive cells were recognized in the lung and the liver. 8. By indirect fluorescent antibody test, the Toxoplasma antibodies in the infected dogs were detected on the 15th day of postinoculation.

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A serological survey of Dirofilaria immitis infection in pet dogs of Busan, Korea, and effects of chemoprophylaxis

  • Byeon, Kang-Hyun;Kim, Bong-Jin;Kim, Sun-Mi;Yu, Hak-Sun;Jeong, Hae-Jin;Ock, Mee-Sun
    • Parasites, Hosts and Diseases
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    • v.45 no.1 s.141
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    • pp.27-32
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    • 2007
  • The status of Dirofilaria immitis infection was assessed in pet dogs of Busan, Korea, and chemoprophylactic effects of microfilaricidal medication were evaluated. A total of 294 pet dogs older than 6 mo were examined, 217 of which had been maintained indoors, and 77 had been kept outdoors. The $Snap^R$ kit and direct microscopic examinations of the peripheral blood were used. The mean overall parasite positive rates were 10.2% and 6.5%, respectively. Outdoor dogs evidenced adult worm infection rate of 31.2% and microfilaria infection rate of 18.2%. The indoor dogs, however, evidenced adult worm infection rate of 2.8% and microfilaria infection rate of 2.3%. The prevalence in males was more than 2 times that of females. The changing pattern of infection rates by age evidenced a gradual increase, from 2- to 6-year-old dogs, after which, a decrease in infection rates was noted. With regard to chemoprophylaxis, the infection rates of complete and incomplete chemoprophylaxis groups were found to be 2-3 times lower than that of the non-chemoprophylaxis group. The results of the present study indicate that the risk of exposure to D. immitis in pet dogs is quite high, particularly in male outdoor dogs, and chemoprophylactic measures were quite effective.

Application of Multiparametric Flow Cytometry (FCM) to Enumerate the Diagnosis of Pseudomonas aeruginosa and Escherichia coli

  • Hwang, Myoung-Goo;Oh, Jung-Woo;Katayama, Hiroyuki;Ohgaki, Shinichiro;Cho, Jin-Kyu
    • Environmental Engineering Research
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    • v.17 no.1
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    • pp.35-39
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    • 2012
  • In this study, multiparametric flow cytometry (FCM) was installed to enumerate the diagnosis of Pseudomonas aeruginosa ATCC 10145 and Escherichia coli K12 (IFO 3301). The nucleic acids (DNA/RNA) were double stained by a LIVE/DEAD bacLight viability kit, involving green SYTO 9 and red propidium iodide (PI), based on the permeability of two chemicals according to the integrity of plasma membrane. As the results showed, the gate for dead bacteria was defined as the range of $0.2{\times}10^0$ to $6.0{\times}10^1$ photo multiplier tube (PMT) 2 fluorescence (X-axis) and $2.0{\times}10^0$ to $2.0{\times}10^2$ PMT 4 fluorescence (Y-axis), and the gate for live bacteria was defined as the range of $6.0{\times}10^0$ to $6.0{\times}10^2$ PMT 2 fluorescence (X-axis) and $2.0{\times}10^0$ to $4.0{\times}10^2$ PMT 4 fluorescence (Y-axis). In the comparison of the number of the tested bacteria detected by FCM (viability assessment) and plate culture (cultivability assessment), the number of bacteria detected by FCM well represented the number of bacteria that was detected by the colony forming unit (CFU) counting method when bacteria were exposed to isopropyl alcohol and silver/copper cations. Consequently, it is concluded that the application of FCM to monitor the functional effect of disinfectants on the physiological status of target bacteria can offer more rapid and reliable data than the plate culture colony counting method.

Changing Proteins in Granulosa Cells during Follicular Development in Pig (돼지 난포 발달 시 과립막 세포에서 발현되는 단백질의 변화)

  • Chae, In-Soon;Jang, Dong-Min;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.33 no.3
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    • pp.183-187
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    • 2009
  • This study analyzed change of proteins in granulosa cells during the porcine follicuar development by proteomics techniques. Granulosa cells of the follicles, of which the diameter is $2{\sim}4\;mm$ and $6{\sim}10\;mm$, were collected from ovary of slaughtered pig that each follicle of diameter $1{\sim}4\;mm$ and $6{\sim}10\;mm$. We extracted glanulosa cell proteins by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $200{\mu}l$. Immobilized pH gradient(IPG) strip used 18 cm, $3{\sim}10\;NL$. SDS-PAGE used 10% acrylamide gel. After silver staining, Melanie 7 and naked eye test were used for spot analyzation. Increasing proteins in glanulosa cell of $6{\sim}10\;mm$ follicle were 7 spots. This spots were analyzed by MALDI-TOF MS and searched on NCBInr. In results, 7 spots were similar to zinc/ling finger protein 3 precursor (RING finger protein 203), angiomotin, heat shock 60 kDa protein 1 (chaperonin) isoform 1 (HSP60), similar to transducin-like enhancer protein 1 (TLE 1), SH3 and PX domains 2A (SH3PXD2A). Those proteins were related with transfer between cells. Increase of proteins has an effect on follicular development.

Cardamonin Inhibited IL-1β Induced Injury by Inhibition of NLRP3 Inflammasome via Activating Nrf2/NQO-1 Signaling Pathway in Chondrocyte

  • Jiang, Jianqing;Cai, Mingsong
    • Journal of Microbiology and Biotechnology
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    • v.31 no.6
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    • pp.794-802
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    • 2021
  • In this study we investigated the role and mechanism of cardamonin on IL-1β induced injury in OA. CHON-001 cells were treated with cardamonin and IL-1β and transfected with silencing nuclear factor erythroid 2-related factor 2 (siNrf2). Cell viability was detected by Cell Counting Kit-8 assay and flow cytometer assay was utilized for cell apoptosis assessment. IL-6, IL-8, TNF-α and Nrf2 mRNA expression was tested by qRT-PCR. Western blot was employed to evaluate MMP-3, MMP-13, Collagen II, Nrf2, NQO-1, NLRP3, Caspase 1 and apoptosis-associated speck-like protein containing a caspase-1 recruitment domain (ASC) protein levels. In CHON-001 cells, IL-1β suppressed cell viability and Collagen II level while promoting cell apoptosis and expression of pro-inflammatory cytokines (IL-6, IL-8, TNF-α), MMPs (MMP-3, MMP-13), NQO-1, and NLRP3 inflammasome (NLRP3, Caspase 1 and ASC), with no significant influence on Nrf2. Cardamonin reversed the effect of IL-1β on cell viability, cell apoptosis, pro-inflammatory cytokines, MMPs, Collagen II, and NLRP3 inflammasome levels. In addition, cardamonin advanced Nrf2 and NQO-1 expression of CHON-001 cells. SiNrf2 reversed the function of cardamonin on IL-1β-induced cell apoptosis and expression of pro-inflammatory cytokines, Nrf2, NQO-1, and NLRP3 inflammasome in chondrocytes. Taken together Cardamonin inhibited IL-1β induced injury by inhibition of NLRP3 inflammasome via activating Nrf2/NQO1 signaling pathway in chondrocyte.

Plasma concentration of dopamine varies depending on breed, sex, and the genotype of DRD4 in horses

  • Junyoung, Kim;Heejun, Jung;Jae-Young, Choi;Jang-Won, Lee;Minjung, Yoon
    • Journal of Animal Science and Technology
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    • v.64 no.4
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    • pp.792-799
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    • 2022
  • Dopamine (DA) is known to be a key modulator of animal behaviors. Thus, the plasma concentration of DA might be used as a biomarker for the behavioral characteristics of horses. The behavioral characteristics of horses vary depending on the breed, age, and sex. Moreover, the DA receptor genotypes are also related to horse behaviors. Thus, the aim of this study was to investigate the DA concentration variations of horse plasma by breed, age, sex, or genotype of its receptor. The horses were divided by breed into Thoroughbred (n = 13), Pony (n = 9), Warmblood (n = 4), and Haflinger (n = 5). The age variable was divided into three different groups: post-pubertal (2-5 years, n = 6), adult (6-13 years, n = 19), and aged horses (15-24 years, n = 6). The sex variable was divided into geldings (n = 8) and mares (n = 23). Approximately 10 mL of blood was collected, and an ELISA kit was used to measure the plasma concentration of DA. Polymerase chain reaction analysis was performed to identify the genetic variation in the DA D4 receptor gene (DRD4). SPSS statistical software was used for statistical analysis. The DA concentrations in geldings were significantly lower than those in mares. There was no significant difference in DA concentrations among breed and age groups. Horses with the GG and GA genotypes had significantly higher plasma concentrations of DA compared to horses with the AA genotype for the G292A gene. Briefly, the plasma concentration of DA varied depending on the sex and genotype of G292A. These factors should be considered when the concentration of DA is used as a biomarker for the behavioral characteristics of horses. In conclusion, the DA concentration or DRD4 genotype of horse plasma has the potential to be used as a biomarker that can predict the behavioral characteristics of horses.

Amygdalin Reverses Macrophage PANoptosis Induced by Drug-Resistant Escherichia coli

  • Xue Yan;Liang Jin;Huifen Zhou;Haofang Wan;Haitong Wan;Jiehong Yang
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1281-1291
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    • 2023
  • Infectious diseases caused by drug-resistant Escherichia coli (E. coli) pose a critical concern for medical institutions as they can lead to high morbidity and mortality rates. In this study, amygdalin exhibited anti-inflammatory and antioxidant activities, as well as other potentials. However, whether it could influence the drug-resistant E. coli-infected cells remained unanswered. Amygdalin was therefore tested in a cellular model in which human macrophages were exposed to resistant E. coli. Apoptosis was measured by flow cytometry and the lactate dehydrogenase (LDH) assay. Western immunoblotting and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) were used to quantify interleukin-18 (IL-18), interleukin-1β (IL-1β), and interleukin-6 (IL-6). The production of reactive oxygen species (ROS) in macrophages was detected by ROS kit. The expression of pan-apoptotic proteins in macrophages was measured by qRT-PCR and Western immunoblotting. Drug-Resistant E. coli inhibited cell viability and enhanced apoptosis in the cellular model. In cells treated with amygdalin, this compound can inhibit cell apoptosis and reduce the expression of pro - inflammatory cytokines such as IL-1β, IL-18 and IL-6. Additionally, it decreases the production of PANoptosis proteins, Furthermore, amygdalin lowered the levels of reactive oxygen species induced by drug-resistant E. coli, in cells, demonstrating its antioxidant effects. Amygdalin, a drug with a protective role, alleviated cell damage caused by drug-resistant E. coli in human macrophages by inhibiting the PANoptosis signaling pathway.