• Title/Summary/Keyword: Gal2

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Increased α2-6 sialylation of endometrial cells contributes to the development of endometriosis

  • Choi, Hee-Jin;Chung, Tae-Wook;Choi, Hee-Jung;Han, Jung Ho;Choi, Jung-Hye;Kim, Cheorl-Ho;Ha, Ki-Tae
    • Experimental and Molecular Medicine
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    • v.50 no.12
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    • pp.9.1-9.12
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    • 2018
  • Endometriosis is a disease characterized by implants of endometrial tissue outside the uterine cavity and is strongly associated with infertility. Focal adhesion of endometrial tissue to the peritoneum is an indication of incipient endometriosis. In this study, we examined the effect of various cytokines that are known to be involved in the pathology of endometriosis on endometrial cell adhesion. Among the investigated cytokines, transforming growth factor-${\beta}1$ ($TGF-{\beta}1$) increased adhesion of endometrial cells to the mesothelium through induction of ${\alpha}2-6$ sialylation. The expression levels of ${\beta}$-galactoside ${\alpha}2-6$ sialyltransferase (ST6Gal) 1 and ST6Gal2 were increased through activation of $TGF-{\beta}RI/SMAD2/3$ signaling in endometrial cells. In addition, we discovered that terminal sialic acid glycan epitopes of endometrial cells engage with sialic acid-binding immunoglobulin-like lectin-9 expressed on mesothelial cell surfaces. Interestingly, in an in vivo mouse endometriosis model, inhibition of endogenous sialic acid binding by a $NeuAc{\alpha}2-6Gal{\beta}1$-4GlcNAc injection diminished $TGF-{\beta}1$-induced formation of endometriosis lesions. Based on these results, we suggest that increased sialylation of endometrial cells by $TGF-{\beta}1$ promotes the attachment of endometrium to the peritoneum, encouraging endometriosis outbreaks.

Lactosylceramide Mediates the Expression of Adhesion Molecules in TNF-${\alpha}$ and IFN ${\gamma}$-stimulated Primary Cultured Astrocytes

  • Lee, Jin-Koo;Kim, Jin-Kyu;Park, Soo-Hyun;Sim, Yun-Beom;Jung, Jun-Sub;Suh, Hong-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.5
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    • pp.251-258
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    • 2011
  • Here we have investigated how lactosylceramide (LacCer) modulates gene expression of adhesion molecules in TNF-${\alpha}$ and IFN ${\gamma}$ (CM)-stimulated astrocytes. We have observed that stimulation of astrocytes with CM increased the gene expression of ICAM-1 and VCAM-1. D-Threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) and N-butyldeoxynojirimycin (NBDNJ), inhibitors of glucosylceramide synthase (GLS) and LacCer synthase (galactosyltransferase, GalT-2), inhibited the gene expression of ICAM-1 and VCAM-1 and activation of their gene promoter induced by CM, which were reversed by exogenously supplied LacCer. Silencing of GalT-2 gene using its antisense oligonucleotides also attenuated CM-induced ICAM-1 and VCAM-1 expression, which were reversed by LacCer. PDMP treatment and silencing of GalT-2 gene significantly reduced CM-induced luciferase activities in NF-${\kappa}B$, AP-1, GAS, and STAT-3 luciferase vectors-transfected cells. In addition, LacCer reversed the inhibition of NF-${\kappa}B$ and STAT-1 luciferase activities by PDMP. Taken together, our results suggest that LacCer may play a crucial role in the expression of ICAM-1 and VCAM-1 via modulating transcription factors, such as NF-${\kappa}B$, AP-1, STAT-1, and STAT-3 in CM-stimulated astrocytes.

Preselection of Bovine Blastocysts Expressing Exogeneous Gene Following Microinjection (외래유전자를 주입한 소 수정란에서 형질전환가능 수정란의 선발)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.167-176
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    • 1997
  • This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

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Characterization of enzymatic activity of galactose epimerase-less mutant of Salmonella pullorum (Galactose epimerase결손 Salmonella pullorum 변이주의 효소활성)

  • Kim, Jong-bae
    • Korean Journal of Veterinary Research
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    • v.34 no.4
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    • pp.781-785
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    • 1994
  • Uridine diphosphate(UDP)-galactose-4-epimerase-less mutants of Salmonella pullorum were isolated after mutagenic treatment with ethidium bromide. When isolated gal E mutants of S. pullorum A2 and D1 were grown in the presence of galactose(0.1 W/V), they exhibited marked bacteriolysis in heart infusion broth. The mutant strains were further investigated the characteristics of enzymatic activities in the Leoloir galactose pathway. Isolated A2 and D1 strains were completely deficient in UDP-galactose-4-epimerase activity. And the activity of other enzymes involved in galactose metabolism were reduced significantly.

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Specificity of Pichia guilliermondii $\alpha$-Galactosidase toward Galactomannans (Galactomannan 이용에 관한 연구 I. Galactomannan에 대한 Pichia guilliermondii유래 $\alpha$-Galactosidase의 특이성)

  • 박귀근
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.5
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    • pp.844-850
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    • 1997
  • $\alpha$-Galactosidase was partially purified from the culture filtrate of pichia guilliermondii by Mannobiose-Sepharose affinity column chromatography. The galactosidase exhibited maximum activity at pH 4.5 and 4$0^{\circ}C$, and was stable in the pH and temperature ranges of 4 to 5.5 and 30 to 6$0^{\circ}C$, respectively. The enzyme was inhibited by $Hg^{2+}$ and $Ag^{2+}$. The enzyme activity was ot affected considerably by treatment with other metal compounds. The enzyme hydrolyzed melibiose to galactose and glucose, raffinose to galactose and sucrose, and $Gal^{3}Man_{3}(6^{3}-$\alpha$-galactosyl-1,4-mannotriose)$ to galactose and mannotriose. On the contrary, it could not hydrolyze $Gal^{3}Man_{4}(6^{3}-galactosyl-1,4-$\alpha$-mannotetraose)$.

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Taxonomical Studies on Discomycetes in Korea(III) (한국산(韓國産) 반균강(盤菌綱)의 분류학적(分類學的) 연구(硏究)(III))

  • Park, Pyoung-Jae;Lee, Ji-Yul;Otani, Yoshio
    • The Korean Journal of Mycology
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    • v.15 no.4
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    • pp.247-249
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    • 1987
  • Twenty three specimens of Discomycetes were collected at Gwangneung, Mt. chiri, Mt. Halla and Mt. Odae from June, 1986 to October, 1987. These Discomycetes were identified and classified into 2 orders, 5 families, 10 genera and 10 species. Among them, Pachyella clypeata(Schw.) Le Gal, Microstoma floccosa(Schw.) Raitviir and Microglossum viride(Pers. ex Fr.) Gill. were newly recorded in Korea. These species were described and their Korean names were designated.

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The effects of manual acupuncture, invasive laser acupuncture and laser skin irradiation at Liver Seunggyeok(肝勝格) on the repair of D-galN-induced Liver Injury in Rats (간승격(肝勝格) 혈위(穴位)에 시술된 침자(鍼刺)와 침습(侵襲) 및 비침습(非侵襲) 레이저침요법(鍼療法)이 간손상(肝損傷) 회복에 미치는 영향)

  • Shin, Hyung-Jin;Kim, Young-Sun;Youn, Dae-Hwan;Lee, Suk-Hee;Oh, Gwang-Hwan;Jeong, Sung-Ho;Na, Chang-Su
    • Korean Journal of Acupuncture
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    • v.27 no.1
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    • pp.49-62
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    • 2010
  • Objective: This study was performed to investigate the effect of manual acupuncture, invasive laser acupuncture and laser skin irradiation at LU8, LR4, HT8 and LR2(Liver Seunggyeok) on D-galN-induced liver injury in rats. Method: Liver injury was induced with D-galN. The experimental rats were divided four groups(Control group, EXP-1, EXP-2, EXP-3). In the Control group, liver injury-induced and not treated. EXP-1 group was liver injury-induced and carried out manual acupuncture with Young-Su(against the meridian course and following the course of the meridian) & Won-Bang(by twisting and rotating the needle) acupuncture method at Liver Seunggyeok. EXP-2 group was liver injury-induced and carried out invasive laser acupuncture at Liver Seunggyeok. EXP-3 group was liver injury-induced and carried out laser skin irradiation at Liver Seunggyeok. Result: In the change of body weight(in 1 week), EXP-1, EXP-2 and EXP-3 groups were significantly increased as compared with control group. In the change AST & ALT, EXP-1 and EXP-2 groups were significantly decreased as compared with control group. In the change of SOD, EXP-1, EXP-2 and EXP-3 groups were significantly increased as compared with control group. In the change of WBC, EXP-2 group was significantly increased as compared with control group. Conclusion: Manual acupuncture, invasive laser acupuncture and laser skin irradiation at Liver Seunggyeok had hepatotherapeutic effect on the treatment of hepatocytotoxity. Invasive laser acupuncture was as effective as manual acupuncture on the treatment of hepatocytotocity.