• Title/Summary/Keyword: BFT

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Bacteroides fragilis Toxin Induces IL-8 Secretion in HT29/C1 Cells through Disruption of E-cadherin Junctions

  • Hwang, Soonjae;Gwon, Sun-Yeong;Kim, Myung Sook;Lee, Seunghyung;Rhee, Ki-Jong
    • IMMUNE NETWORK
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    • v.13 no.5
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    • pp.213-217
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    • 2013
  • Enterotoxigenic Bacteroides fragilis (ETBF) is a human gut commensal bacteria that causes inflammatory diarrhea and colitis. ETBF also promotes colorectal tumorigenesis in the Min mouse model. The key virulence factor is a secreted metalloprotease called B. fragilis toxin (BFT). BFT induces E-cadherin cleavage, cell rounding, activation of the ${\beta}$-catenin pathway and secretion of IL-8 in colonic epithelial cells. However, the precise mechanism by which these processes occur and how these processes are interrelated is still unclear. E-cadherin form homophilic interactions which tethers adjacent cells. Loss of E-cadherin results in detachment of adjacent cells. Prior studies have suggested that BFT induces IL-8 expression by inducing E-cadherin cleavage; cells that do not express E-cadherin do not secrete IL-8 in response to BFT. In the current study, we found that HT29/C1cells treated with dilute trypsin solution induced E-cadherin degradation and IL-8 secretion, consistent with the hypothesis that E-cadherin cleavage causes IL-8 secretion. However, physical damage to the cell monolayer did not induce IL-8 secretion. We also show that EDTA-mediated disruption of E-cadherin interactions without E-cadherin degradation was sufficient to induce IL-8 secretion. Finally, we determined that HT29/C1 cells treated with LiCl (${\beta}$-catenin activator) induced IL-8 secretion in a dose-dependent and time-dependent manner. Taken together, our results suggest that BFT induced IL-8 secretion may occur by the following process: E-cadherin cleavage, disruption of cellular interactions, activation of the ${\beta}$-catenin pathway and IL-8 expression. However, we further propose that E-cadherin cleavage per se may not be required for BFT induced IL-8 secretion.

Study on Growth Curves of Longissimus dorsi Muscle Area, Backfat Thickness and Body Conformation for Hanwoo (Korean Native) Cows

  • Lee, J.H.;Oh, S.H.;Lee, Y.M.;Kim, Y.S.;Son, H.J.;Jeong, D.J.;Whitley, N.C.;Kim, J.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.9
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    • pp.1250-1253
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    • 2014
  • The objective of this study was to estimate the parameters of Gompertz growth curves with the measurements of body conformation, real-time ultrasound longissimus dorsi muscle area (LMA) and backfat thickness (BFT) in Hanwoo cows. The Hanwoo cows (n = 3,373) were born in 97 Hanwoo commercial farms in the 17 cities or counties of Gyeongbuk province, Korea, between 2000 and 2007. A total of 5,504 ultrasound measurements were collected for the cows at the age of 13 to 165 months in 2007 and 2008. Wither height (HW), rump height (HR), the horizontal distance between the top of the hips (WH), and girth of chest (GC) were also measured. Analysis of variance was conducted to investigate variables affecting LMA and BFT. The effect of farm nested in location was included in the statistical model, as well as the effects of HW, HR, WH, and GC as covariates. All of the effects were significant in the analysis of variance for LMA and BFT (p<0.01), except for the HR effect for LMA. The two ultrasound measures and the four body conformation traits were fitted to a Gompertz growth curve function to estimate parameters. Upper asymptotic weights were estimated as $54.0cm^2$, 7.67 mm, 125.6 cm, 126.4 cm, 29.3 cm, and 184.1 cm, for LMA, BFT, HW, HR, WH, and GC, respectively. Results of ultrasound measurements showed that Hanwoo cows had smaller LMA and greater BFT than other western cattle breeds, suggesting that care must be taken to select for thick BFT rather than an increase of only beef yield. More ultrasound records per cow are needed to get accurate estimates of growth curve, which, thus, helps producers select animals with high accuracy.

The Relationships of Plasma Leptin, Backfat Thickness and TDN Intake across Finishing Stage of Holstein Steers

  • Vega, R.A.;Hidari, H.;Kuwayama, H.;Suzuki, M.;Manalo, D.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.3
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    • pp.330-336
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    • 2004
  • Six 16 months old Holstein steers were offered ad libitum feed for 7 months, to determine the (1) relationships of backfat thickness (BFT) to plasma leptin, and insulin; and (2) associations of TDN intake/kg body weight (BW) to plasma leptin, BFT and insulin. Feed intake, body weight and BFT were measured on selected monthly ages from day 1 to 8, day 1 and 8, and day 8, respectively. Blood was sampled on day 8 and the plasma was analyzed for leptin, insulin, glucose, NEFA, total cholesterol and triglyceride. Body weight and BFT increased, while TDN intake per kg BW decreased from 16 to 23 months old. Plasma leptin increased and mimicked the level of insulin, resulting to significant correlation (r=0.54; p<0.002). TDN intake was negatively related to plasma leptin (r=0.49; p<0.004), insulin (r=0.41; p<0.02) and BFT at 12 to 13th rib (r=0.48; p<0.005). Backfat thickness at 12 to 13th rib was positively related to plasma leptin (r=0.45; p<0.01). Negative associations of TDN intake with plasma leptin and BFT during finishing period suggest long-term involvement of adipose tissues in the feed intake regulation of steers fed high concentrate diet.

Cloning and Expression of the Gene Encoding Glucose Permease of the Phosphotransferase System from Brevibacterium flavum in Escherichia coli

  • Kwon, Il;Lee, Kyu-Nam;Lee, Jung-Kee;Pan, Jae-Gu;Oh, Tae-Kwang;Lee, Hyung-Hoan;Yoon, Ki-Hong
    • Journal of Microbiology and Biotechnology
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    • v.5 no.4
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    • pp.188-193
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    • 1995
  • A Brevibacterium flavum gene coding for glucose permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing the Escherichia coli ZSCl13 mutations affecting a ptsG gene with the B. flavum genomic library. From the E. coli clone grown as red colony on a MacConkey plate supplemented with glucose as an additional carbon source, a recombinant plasmid was isolated and named pBFT93. The plasmid pBFT93 was identified as carrying a 3.6-kb fragment of B. flavum chromosomal DNA which enables the E. coli transformant to use glucose or man nose as a sole carbon source in an M9 minimal medium. The non-metabolizable sugar analogues, 2-deoxy-D-glucose (2-DG) and methyl-$\alpha$-D-glucopyranoside (MeGlc) affected the growth of ZSCl13 cells carrying the plasmid pBFT93 on minimal medium supplemented with non-PTS carbohydrate, glycerol, as a sole cabon source, while the analogues did not repress the growth of ZSCl13 cells without pBFT93. It was also found that both $2-deoxy-D-[U-^{14}C]glucose{\;}and{\;}methyl-{\alpha}-D-[U-^{14}C]glucopyranoside$ could be effectively transported into ZSCl13 cells transformed with plasmid pBFT93. Several in vivo complementation studies suggested that the B. flavum DNA in pBFT93 encodes a glucose permease specific for glucose and mannose.

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Monitoring of Pacific Whiteleg Shrimp Litopenaeus vannamei Pathogens Cultured with Biofloc Technology on the West Coast of Korea, 2021 (2021년 서해권역 실내 바이오플락 양식기술(Bioflocs Technology)로 사육한 흰다리새우(Litopenaeus vannamei) 병원체 모니터링)

  • Hyun Jung Gye;Su-kyoung Kim;Hee Woong Kang;Hyun Mi Jung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.1
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    • pp.133-139
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    • 2023
  • The advantage of biofloc technology (BFT) in aquaculture is in the prevention of pathogenic transmission. In this study, we performed an investigation on viral, bacterial, and microsporidian parasite infections targeting a total of 194 whiteleg shrimp Litopenaeus vannamei reared in seven BFT-farms on the west coast of Korea in 2021. Hepatopancreatic and cuticular epithelium and pereiopods tissues of shrimp were tested for the four pathogens, Enterocytozoon hepatopenaei (EHP), Vibrio parahaemolyticus causing Acute Hepatopancreatic necrosis disease (VPAHPND), white spot syndrome virus (WSSV), and hepatopancreatic parvovirus (HPV). The microsporidian parasite EHP was detected in the hepatopancreatic tissue of BFT whiteleg shrimp in the Ganghwa region, whereas no other pathogenic bacteria or virus was detected on the shrimp in the seven BFT-farms. As a result of bacterial flora in the rearing water of BFT whiteleg shrimp using DNA microbiome technology, V. chemaguriensis and V. alfacsensis were contained at 0.05% and 0.01%, respectively, but no VPAHPND was detected. These findings will serve as a basis for supporting safe BFT-aquaculture of whiteleg shrimp.

Genetic Analyses of Carcass Characteristics in Crossbred Pigs: Cross between Landrace Sows and Korean Wild Boars

  • Choy, Y.H.;Jeon, G.J.;Kim, T.H.;Choi, B.H.;Cheong, I.C.;Lee, H.K.;Seo, K.S.;Kim, S.D.;Park, Y.I.;Chung, H.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.8
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    • pp.1080-1084
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    • 2002
  • Carcass characteristics of 241 crossbred pigs (Korean wild boars ${\times}$ Landrace sows) were analyzed to examine variations in fasted body weight (FASTWT), carcass weight (CARCWT), dressing percentage (DP), back fat thickness (BFT) and longissimus muscle weight (LMW), and to estimate genetic and phenotypic parameters using three different slaughter-end points. Covariates in the least squares full sib model were slaughter age, fasted body weight and back fat thickness of the carcass. Coefficient of variation was highest for BFT followed by LMW, CARCWT, FASTWT and DP in magnitude. Regressions of three covariates on traits were all linear. However, slaughter age was not significant as a linear covariate for five traits while FASTWT was significant for CARCWT and LMW and BFT was significant for all remaining traits. Genetic and phenotypic variation was considerably reduced by regressing FASTWT or BFT in the model. Heritability estimates of FASTWT, CARCWT, DP and BFT were 0.68, 0.61, 0.11 and 0.49, respectively, using slaughter age as covariate (model 1). Those of CARCWT, DP, BFT and LMW were 0.15, 0.15, 0.30 and 0.11, respectively, using FASTWT as covariate (model 2). Heritability estimates of the traits using LMW as covariate (model 3) were similar to the estimates from Model 1 except that the estimate of CARCWT was reduced to 0.39. Genetic or phenotypic correlations among FASTWT, CARCWT and BFT were all positive and moderate to high. Those between BFT and LMW were also positive and low to moderate. However, genetic and phenotypic correlations between DP and CARCWT were positive while those between DP and FASTWT were negative. It was suggested from this study that differences in carcass yield traits be determined using slaughter age or back fat thickness as slaughter-end point and carcass quality traits using fasted body weight as slaughter-end point.

Bacteroides fragilis Toxin Induces Cleavage and Proteasome Degradation of E-cadherin in Human Breast Cancer Cell Lines BT-474 and MCF7 (인간 유방암 세포주 BT-474와 MCF7에서 Bacteroides fragilis Toxin에 의한 E-cadherin 분절과 프로테아좀에 의한 분해)

  • Da-Hye KANG;Sang-Hyeon YOO;Ju-Eun HONG;Ki-Jong RHEE
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.1
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    • pp.37-44
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    • 2023
  • Enterotoxigenic Bacteroides fragilis (ETBF) has been reported to promote colitis and colon cancer through the secretion of B. fragilis toxin (BFT), a zinc-dependent metalloprotease. In colonic epithelial cells, BFT induces the cleavage of E-cadherin into the 80 kDa ectodomain and the 33 kDa membrane-bound intracellular domain. The resulting membrane-tethered fragment is then cleaved by γ-secretase forming the 28 kDa E-cadherin intracellular fragment. The 28 kDa cytoplasmic fragment is then degraded by an unknown mechanism. In this study, we found that the 28 kDa E-cadherin intracellular fragment was degraded by the proteasome complex. In addition, we found that this sequential E-cadherin cleavage mechanism is found not only in colonic epithelial cells but also in the human breast cancer cell line, BT-474. Finally, we report that staurosporine also induces E-cadherin cleavage in the human breast cancer cell line, MCF7, through γ-secretase. However, further degradation of the 28 kDa E-cadherin intracellular domain is not dependent on the proteasome complex. These results suggest that the BFT-induced E-cadherin cleavage mechanism is conserved in both colonic and breast cancer cells. This observation indicates that ETBF may also play a role in the carcinogenesis of tissues other than the colon.

Biceps Femoris Tendon and Lateral Collateral Ligament: Analysis of Insertion Pattern Using MRI (대퇴이두건과 외측 측부인대: 자기공명영상을 이용한 부착형태 유형의 분석)

  • Shin, Yun Kyung;Ryu, Kyung Nam;Park, Ji Seon;Lee, Jung Eun;Jin, Wook;Park, So Young;Yoon, So Hee;Lee, Kyung Ryeol
    • Investigative Magnetic Resonance Imaging
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    • v.18 no.3
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    • pp.225-231
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    • 2014
  • Purpose : The biceps femoris tendon (BFT) and lateral collateral ligament (LCL) in the knee were formerly known to form a conjoined tendon at the fibular attachment site. However, the BFT and LCL are attached into the fibular head in various patterns. We classified insertion patterns of the BFT and LCL using MR imaging, and analyzed whether the LCL attaches to the fibular head or not. Materials and Methods: A total of 494 consecutive knee MRIs of 470 patients taken between July 2012 and December 2012 were retrospectively reviewed. There were 224 males and 246 females, and patient age varied from 10 to 88 (mean, 48.6). The exclusion criteria were previous surgery and poor image quality. Using 3T fat-suppressed proton density-weighted axial images, the fibular insertion patterns of the BFT and LCL were classified into following types: type I (the LCL passes between the anterior arm and direct arm of the BFT's long head), type II (the LCL joins with anterior arm of the long head of the BFT), type III (the BFT and LCL join to form a conjoined tendon), type IV (the LCL passes laterally around the anterior margin of the BFT), and type V (the LCL passes posteriorly to the direct arm of the BFT's long head). Results: Among the 494 cases of the knee MRI, there were 433 (87.65%) type I cases, 21 (4.25%) type II cases, 2 (0.4%) type III cases, 16 (3.23%) type IV cases, and 22 (4.45%) type V cases. There were 26 cases (5.26%) in which the LCL and BFT were not attached into the fibular head. Conclusion: The fibular attachment pattern of the BFT and LCL shows diverse types in MR imaging. The LCL does not adhere to the head in some patients.

Enterotoxigenic Bacteroides fragilis-Associated Diseases and Detection (Enterotoxigenic Bacteroides fragilis에 의한 질환과 검출)

  • Gwon, Sun-Yeong;Jang, In-Ho;Rhee, Ki-Jong
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.161-167
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    • 2015
  • These commensal intestinal bacteria can enhance the immune system and aid in nutrient absorption but can also act as opportunistic pathogens. Among these intestinal bacteria, the anaerobic Bacteroides fragilis are divided into enterotoxigenic B. fragilis (ETBF) which secrete the B. fragilis toxin (BFT) and non-enterotoxigenic B. fragilis (NTBF) which do not secrete BFT. ETBF can cause diarrhea and colitis in both humans and livestock but can also be found in asymptomatic individuals. ETBF is predominantly found in patients with inflammatory diarrheal diseases and traveller's diarrhea. Several clinical studies have also reported an increased prevalence of ETBF in human patients with inflammatory bowel disease (IBD), colitis and colorectal cancer. In small animal models (C57BL/6 wild-type mice, germ-free mice, multiple intestinal neoplasia (Min) mice, rabbits and Mongolian gerbils), ETBF have been found to initiate and/or aggravate IBD, colitis and colorectal cancer. BFT induces E-cadherin cleavage in intestinal epithelial cells resulting in loss of epithelial cell integrity. Subsequent activation of the ${\beta}$-catenin pathway leads to increased cellular proliferation. In addition, ETBF causes acute and chronic colitis in wild-type mice as well as enhances tumorigenesis in Min mice via activation of the Stat3/Th17 pathway. Currently, ETBF can be detected using a BFT toxin bioassay and by PCR. Advances in molecular biological techniques such as real-time PCR have allowed both researchers as well as clinicians to rapidly detect ETBF in clinical samples. The emergence of more sensitive techniques will likely advance molecular insight into the role of ETBF in colitis and cancer.

Cloning, Sequencing, and Characterization of Enterotoxin Pathogenicity Islet from Bacteroides fragilis 419

  • Rhie, Gi-Eun;Chung, Gyung-Tae;Lee, Yong-Jin;Sung, Won-Keun;Oh, Hee-Bok
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.86-90
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    • 2000
  • We have earlier reported on the cloning and identification of bft-k from an enterotoxigenic strain of Bacteroides fragilis 419, which was isolated from the blood of a Korean patient who suffered from systemic infections [4,5]. The bft-k gene encodes a 397-amino-acids metalloprotease enterotoxin, and the protein has been identified as a new isoform of B. fragilis enterotoxins (BFTs), which are cytopathic to intestinal epithelial cells to induce fluid secretion and tissue damage in ligated intestinal loops [4, 6, 18, 20]. This report describes the cloning and sequencing of the enterotoxin pahogenicity islet of B. fragilis 419 which contains the bft-k gene. the cloned enterotoxin pathogenicity islet was found to have 6,045 bp in length and to contain 120bp direct repeats near its end. In the pathogenicity islet, in addition to the BFR-K, two putative open reading frames (ORFs) were identified; (1) the t-3 gene encoding a 396-amino-acids protein of a putative metalloprotease; (2) the third gene encoding an ORF of a 59-amino-acids protein, whose function has not yet beenn characterized. The expression of the t-3 gene in B. fragilis 419 was verified by western blot analysis.

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