• Title/Summary/Keyword: binding treatment

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Effect of trichostatin A on NF-κB DNA binding activity and myogenesis in C2Cl2 skeletal muscle Precursor cell (C2C12 근육아세포에서 trichostatin A에 의한 NF-κB DNA 결합 활성과 근육발생에 미치는 영향)

  • 임운기;김경창;신혜자
    • Journal of Life Science
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    • v.12 no.1
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    • pp.55-60
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    • 2002
  • The differentiation of skeletal muscle precursor cells in culture is marked by the transcriptional activation of muscle-specific genes and the morphological differentiation of myoblast into multinucleate myotube. In this study, we examined the effect of TSA (Trichostatin A) on WF-kB DNA binding activity and muscle cell fusion in the process of myogenesis. Under TSA treatment, C2C12 myoblast could not fuse to myotube and its NF-kB DNA binding activity was also blocked. To investigate whether these phenomenons were affected by TSA in direct or not, differentiation media (DM) used to differentiate cells without TSA was concentrated and added to C2C12 myoblast with TSA simultaneously. C2C12 myoblast was fused to myotube and NF-kB DNA binding activity was recovered. These results suggest that TSA affects on the differentiation of myoblast, perhaps through several factors, by inhibiting myoblst fusion and blocking NF-kB DNA binding activity.

Study on the Detection of Anti-Sperm Antibodies in Zygote Intra Fallopian Transfer (ZIFT) Patients with Fertilization Failure or Low Fertilization Rate and Retreatment of Oocyte and Sperm (접합자 난관내 이식 환자에 있어서 수정 실패와 항정자 항체와의 관계 및 난자와 정자의 재처리에 관한 연구)

  • Chung, M.K.;Ko, J.J.;Do, B.R.;Koo, J.J.;Han, S.Y.;Cha, K.Y.
    • Clinical and Experimental Reproductive Medicine
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    • v.19 no.2
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    • pp.169-174
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    • 1992
  • Previous studies have indicated that immunological factor is responsible for the infertility. We have detected sperm antibodies in ZIFT patients which grouped as fertilization failure (A; n=18) and low fertilization rate (${\leq}50%$)(B; n=20). Patients, however, had normal oocytes and sperms. We collected serum from wives and semen from husbands and donors (fertile sperm), if it was needed. We examined class, binding patterns and amounts of antisperm antibodies(ASA) by direct and indirect immunobead binding assay. In group A, 11 husbands were ASA positive showing 62.2% and 61.1% binding with IgA and IgG, respectively, and two wives were ASA positive showing 70.0% and 71.0% binding with IgA and IgG, respectively. Binding sites were mainly at the head of sperms (84%). In group B, 8 husbands were ASA positive showing 37.5% and 40.0% binding with IgA and IgG, respectively, and two wives were ASA positive showing 41.3% and 42.0% binding with IgA and IgG, respectively. Binding sites were also mainly at the head of sperms (78%). For the treatment of ZIFT patients who had fertilization failure at the first trial, we used albumin fractionation method and dilution method with 30% fetal cord serum (FCS) to reduce the titer of ASA. We used partial zona dissection (P.Z.D.) method for wives who have antisperm antibodies in their serum. According to represented method, we could inhance the fertilization rate to 60.0% by albumin fractionation and 20.0% by P.Z.D., respectively. We concluded that the use of micromanipulation like P.Z.D. or the other sperm processing methods is required to increase a chance of fertilization. This result suggested that it should be a prerequisite to test antisperm antibodies prior to entering assisted reproductive technologies (ART) programs.

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A Study on the Mechanism of Calcium Binding Inhibition of Cardiac Sarcoplasmic Reticulum by Oxygen Free Radicals (산소대사물에 의한 심장근 Sarcoplasmic reticulum의 칼슘운반 억제 기전에 관한연구)

  • Kim, Hae-Won;Chung, Myung-Hee;Kim, Myung-Suk;Park, Chan-Woong
    • The Korean Journal of Pharmacology
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    • v.21 no.2
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    • pp.79-89
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    • 1985
  • Mechanism of calcium transport inhibition of cardiac sarcoplasmic reticulum (SR) by oxygen free radicals was examined. Effects of oxygen free radicals generated by xanthine/xanthine oxidase (X/XO) system on isolated porcine ventricle SR were studied with respect to its calcium binding, lipid peroxidation, SH-group content and alteration of membrane protein components. The results are as follows. 1) Calcium binding of isolated SR was markedly inhibited by X/XO. 2) During the incubation of sarcoplasmic reticulum with xanthine/xanthine oxidase, there were marked inclose in lipid peroxidation and reduction of SH-group content. 3) An antioxidant, p-phenylenediamine effectively prevented the lipid peroxidation but partially prevented the calcium binding inhibition of X/XO treated SR. 4) The reduction of SH-group content of SR treated with X/XO was partially prevented by p-phenylendiamine. 5) When modifying SH-group of SR by treatment with DTNB, the inhibition of calcium binding activity was partially prevented. 6) On gel-permeation chromatography of X/XO-treated sarcoplasmic reticulum, there was an increase of small molecular weight products, probably protein degradation products. 7) Semicarbazide, which prevents the cross-linking reaction of protein components, did not affect the calcium binding inhibition of X/XO-treated SR. From these results, it is suggested that the inhibition of calcium binding of SR by oxygen free radicals results from the consequence of multiple changes of SR components, which are lipid peroxidation, SH-group oxidation and degradation of protein components.

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Effects of Salt, Glucono-${\delta}$-Lactone and High Pressure Treatment on Physico-Chemical Properties of Restructured Pork (소금과 Glucono-${\delta}$-Lactone의 첨가 및 초고압 처리가 재구성 돈육의 이화학적 특성에 미치는 효과)

  • Bong Geun-Pyo;Park Sung-Hee;Kim Jee-Yeon;Ko Se-Hee;Min Sang-Gi
    • Food Science of Animal Resources
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    • v.26 no.2
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    • pp.204-211
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    • 2006
  • This study investigated the effect of salt and glucono-${\delta}$-lactone (GdL) on the cold-set binding of restructured pork washed and pressurized at 200 MPa. Binding strength, PH, water holding capacity (WHC) and color were determined. NaCl improved pH, WHC and binding strength. GdL also increased binding strength while decreased WHC and pH significantly (p<0.05). However, low GdL level combined with NaCl showed high pH and WHC, compared to control. In color, NaCl decreased $L^*$-value with increasing $a^*$-value significantly (p<0.05). In contrast to NaCl, GdL increased $L^*$-value and decreased $a^*$-value. GdL tended to decrease $b^*$-value and significant differences were found when GdL was added above 1%. Pearson's correlation coefficients presented that NaCl had a significant effect on binding strength (0.6632) and lightness (-0.7330) while GdL had a significant correlation with all parameters barring binding strength. The results indicated that under washing and pressure treatments, GdL had a potential effect on cold-set binding with reducing NaCl concentration, especially when low GdL concentration combined with NaCl was added.

Inhibition of Adipocyte Differentiation and Adipogenesis by Supercritical Fluid Extracts and Marc from Cinnamomum verum (초임계 추출 계피오일의 3T3-L1 지방전구세포의 분화 전사인자 억제에 의한 지방대사 조절)

  • Park, Sung-Jin;Lee, In-Seon;Lee, Sam-Pin;Yu, Mi-Hee
    • Journal of Life Science
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    • v.23 no.4
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    • pp.510-517
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    • 2013
  • This study was performed to evaluate the antiobesity effect of supercritical fluid extracts (SFC) and marc methanol extracts (SFM) from Cinnamomum verum in 3T3-L1 preadipocytes. In inducing the differentiation of 3T3-L1 preadipocytes in the presence of an adipogenic cocktail, iso-butylmethylanthine (IBMX), dexamathasone, and insulin, treatment with fraction residue SFC and SFM. SFC significantly reduced the mRNA expression of the transcription factor peroxisome proliferator-activate-dreceptor-${\gamma}$ ($PPAR{\gamma}$), the sterol regulatory-element-binding protein-1c (SREBP1c), and the CCAAT enhancer-binding-protein ${\alpha}$ ($C/EBP{\alpha}$) in a concentration-dependent manner. Moreover, SFC markedly down-regulated acyl-CoA synthetase-1 (ASC1), fatty acid synthesis (FAS), fatty acid transport-1 (FATP1), fatty acid binding protein 4 (FABP4), and perilipin. These findings suggest that SFC may be a potential therapeutic adjunct for obesity by targeting the differentiation of preadipocytes, as well as their functions.

A Study on the Diurnal Variation of Endorphin in Rat Brain (백서 뇌내 Endorphin의 일주기 변동에 관한 연구)

  • Jung, Chang-Young;Kim, Ki-Won;Cho, Kyu-Park
    • The Korean Journal of Pharmacology
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    • v.20 no.2
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    • pp.35-47
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    • 1984
  • Contents of immunoreactive ${\beta}-endorphin$ and maximum of $^3H-morphine$ binding was measured in the rat midbrain homogenates from different subgroups at 24 hour interval over 24 hours. Animals were adapted to the light-dark cycle(L : D, 12: 12) or constant darkness (D : D, 12 : 12) for 3 weeks. After the adaptation, 0.5ml of physiologic saline or drug was administered twice a day for 2 weeks. A highly significant circadian rhythm with the peak$(94.8{\pm}7.7\;fmole/mg\;protein)$ at 06:00 and the nadir $(27.6{\pm}2.4\;fmole/mg\;protein)$ at 18:00 was observed in constant of group. Constant dark or treatment of reserpine, pargyline, imipramine, amphetamine and chlorpromazine modified the diurnal rhythm in the time of peak and nadir, shape, phase amplitude and 24 hour mean of ${\beta}-endorphin$ contents. Opiate receptor binding by $^3H-morphine$ also showed highly significant diurnal change in control and constant dark adapted rats. Statistical analysis by one-way analysis of variance and two-way analysis of variance indicates that the·re are highly significant differences between the diurnal change of ${\beta}-endorphin$ in control and those constant dark adapted and drug treated groups. However diurnal change of Maximum $^3H-morphine$ binding is closely related to the change of ${\beta}-endorphin$ contents. The results are interpreted with regard to the circadian rhythm of beta-endorphin contents, its modification by psychoactive drugs and possible mechanism of diurnal change of opiate receptor in brain.

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Effects of Safflower Seeds on the Serum Levels of Insulin-like Growth Factors, Insulin-like Growth Factor Binding Protein-3 and BALP in Osteoporosis Induced-ovariectomized Rats (흰쥐의 난소제거로 유발한 골다공증에 대한 홍화씨의 IGFs, IGF binding protein-3 그리고 BALP에 대한 혈청내 효과)

  • Kim, Soo-mi;Park, In-hyuk;Kim, Nam-soo
    • Journal of Veterinary Clinics
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    • v.20 no.3
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    • pp.263-273
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    • 2003
  • This study was carried out to investigate the effects of the Korean Safflower (Carthamus inctorius L) seed powder on serum level of hormones and trabecula area during the recovery from osteoporosis induced ovariectomized rats. Four month-old rats were ovariectomized (OVX), remained untreated for 8 weeks, and were subsequently administered safflower seed (0.03 g/kg) every other day 30 for days. We examined the effects of treated safflower seed every 10 days on ovariectomy-related changes in Insulin-like Growth Factors, Insulin-like Growth Factor binding protein-3 (IGFBP-3), Estrogen, Bone-specific alkaline phosphotase, Calcium, and Phospotase in the serum, and also histomorphology of the proximal fibula metaphysis and femur/body weight rate. Ten and 20 days after safflower seed treatment in OVX rats, serum levels of IGF-I, -II and IGFBP-3 were not different from the Sham and OVX groups. In 30 days, serum levels of IGF-I,-II and IGFBP-3 were higher after safflower seed treatment in OVX rats as compared to the other two groups (p<0.05). Bone alkaline phosphatase levels were increased through safflower seed treatment in OVX rats compared to the other two groups in 30 days. There were no differences between OVX and safflower seed treated OVX rats in serum levels of estrogen and femur/body weight rate, but estrogen levels for the sham group were higher than for the other two groups. The safflower seed is increased to serum levels of IGFs, IGFBP-3 and BALP of osteoporosis induced by ovariectomized rats. Thus, we conclude that the safflower seed is a possible role for improvement of osteoporosis induced-ovariectomized rats.

Negative Regulation of Tumor Suppressor p53 at the Promoter Regions of Oncogenic SETDB1 and FosB Genes (암종양유전자 SETDB1과 FosB 발현에 대한 p53의 음성 조절기작)

  • Yun, Hyeon Ji;Na, Han-Heom;Kim, Keun-Cheol
    • Journal of Life Science
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    • v.30 no.12
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    • pp.1070-1077
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    • 2020
  • Treatment with anticancer drugs changes the expression of multiple genes related to cell proliferation, migration, and drug resistance. These changes in gene expression may be connected to regulatory networks for each other. This study showed that doxorubicin treatment induces the expression of oncogenic FosB and decreases the expression of oncogenic SETDB1 in A549 and H1299 human lung cancer cells, which are different in tumor suppressor p53 status. However, a small difference was detected in the quantitative expression of those proteins in the two kinds of cells. To examine the potential regulation of SETDB1 and FosB by p53, we predicted putative p53 binding sites on the genomic DNA of SETDB1 and FosB using a TF motif binding search program. These putative p53 binding sites were identified as 18 sites in the promoter regions of SETDB1 and 21 sites in the genomic DNA of FosB. A luciferase assay confirmed that p53 negatively regulated the promoter activities of SETDB1 and FosB. Furthermore, the results of RT-PCR, western blot, qPCR, and immunostaining experiments indicated that the transfection of exogenous p53 decreases the expression of SETDB1 and FosB in H1299 cells. This indicates that p53 negatively regulates the expression of SETDB1 and FosB at the transcriptional level. Collectively, the downregulation of SETDB1 and FosB by p53 may provide functional networks for apoptosis and for the survival of cancer cells during anticancer drug treatment.

Microarray analysis of gene expression in raw cells treated with scolopendrae corpus herbal-acupuncture solution (蜈蚣(오공) 약침액(藥鍼液)이 LPS로 처리된 RAW 세포주(細胞柱)의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響))

  • Bae, Eun-Hee;Lee, Kyung-Min;Lee, Bong-Hyo;Lim, Seong-Chul;Jung, Tae-Young;Seo, Jung-Chul
    • Korean Journal of Acupuncture
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    • v.23 no.3
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    • pp.133-160
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    • 2006
  • Objectives : Scolopendrae Corpus has a broad array of clinical applications in Korean medicine, including treatment of inflammatory conditions such as arthritis. To explore the global gene expression profiles in human Raw cell lines treated with Scolopendrae Corpus herbal-acupuncture solution (SCHAS), cDNA microarray analysis was performed. Methods : The Raw 264.7 cells were treated with lipopolysaccharide (LPS), SCHAS, or both. The primary data was normalized by the total spots of intensity between two groups, and then normalized by the intensity ratio of reference genes such as housekeeping genes in both groups. The expression ratio was converted to log2 ratio. Normalized spot intensities were calculated into gene expression ratios between the control and treatment groups. Greater than 2 fold changes between two groups were considered to be of significance. Results : Of the 8 K genes profiled in this study, with a cut-off level of two-fold change in the expression, 20 genes (BCL2-related protein A1, MARCKS-like 1, etc.) were upregulated and 5 genes (activated RNA polymerase II transcription cofactor 4, calcium binding atopy-related autoantigen 1, etc.) downregulated following LPS treatment. 139 genes (kell blood group precursor (McLeod phenotype), ribosomal protein S7, etc.) were upregulated and 42 genes (anterior gradient 2 homolog (xenopus laevis), phosphodiesterase 8B, etc.) were downregulated following SCHAS treatment. And 10 genes (yeast saccharomyces cerevisiae intergeneic sequence 4-1, mitogen-activated protein kinase 1, etc.) were upregulated and 8 genes (spermatid perinuclear RNA binding protein, nuclear receptor binding protein 2, etc.) were downregulated following co-stimulation of SCHAS and LPS. Discussions : It is thought that microarrays will play an ever-growing role in the advance of our understanding of the pharmacological actions of SCHAS in the treatment of arthritis. But further studies are required to concretely prove the effectiveness of SCHAS.

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Reevaluation of the Effect of Phenobarbital on the Response to Pain in Rat (백서에서 동통에 미치는 Phenobarbital 효과의 재평가)

  • So, Byoung-Gyoum;Kim, Kee-Won;Ko, Myoung-Kyu;Yang, Won-Mo;Cho, Kyu-Park
    • The Korean Journal of Pharmacology
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    • v.22 no.2
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    • pp.88-95
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    • 1986
  • Clinically, subhypnotic doses of barbiturates have been known to elicit hyperalgesia. In this experiment, effect of acute or chronic phenobarital treatment on the response to pain in rat was reevaluated by hot-plate method. To elucidate its mechanism, changes of ${\beta}-endorphin$ contents and [3H]-morphine binding of the rat midbrain as well as functional opiate receptor in vas deferens were also measured. Intraperitoneal injection of sub anesthetic dose phenobarbital induced initial hyperalgesia followed by successive analgesia, while chronic phenobarbital-treatment decreased reactivity to pain. Naloxone (10mg/kg, i.p.) markedly shortened hot plate latency period, and significantly inhibited the analgesic action of phenobarbital. Single dose of phenobarbital did not affect ${\beta}-endorphin$ contents and [3H]-morphine binding in rat mid brain, but in the chronic phenobarbital-treated groups, ${\beta}-endorphin$ contents was increased, while Bmax of opiate receptor binding was decreased. Moreover, very significant correlations among responses to pain, changes of ${\beta}-endorphin$ contents and opiate receptor binding were observed. However, Kd values of opiate receptor bindings were not changed in all preparations. In the chronic phenobarbital-treated vas deferens preparations, ID50 of morphine was increased witb concomittant decrease of maximum effect. But $pA_2 $, value for naloxone was not changed. From these results, it is suggested that phenobarbital can produce analgesia due to changes of ${\beta}-endorphin$ contents as well as functional opiate receptors by receptor regulation.

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