• Title/Summary/Keyword: pH stimulation

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Interaction of $17{\beta}-Estradiol$ with EGF and IGF-I on Proliferation and $P_i$ Uptake in Primary Cultured Rabbit Renal Proximal Tubular Cells

  • Han, Ho-Jae;Lee, Yeun-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.4
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    • pp.493-501
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    • 1998
  • The most significant direct role of estrogen in vivo is its ability to elicit receptor-mediated cellular proliferation in mammalian target tissues. However, the mechanism by which exogenously added estrogen causes the neoplastic transformation of renal cortical cells is yet to be uncovered. The present study was designed to evaluate interaction of $17{\beta}-estradiol\;(E_2)$ with epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) on proliferation and $P_i$ uptake in primary cultured rabbit renal proximal tubular cells in phenol red-free, hormonally defined-medium. $[^3H]-thymidine$ incorporation increased markedly by about 133% and 141% more in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$, respectively, than that of control. Cell count was 162% and 143% greater in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$ , respectively, compared with control. Among all time points examined, there was an increase in $[^3H]-thymidine$ incorporation in the presence of $10^{-9}\;M\;E_2$ at day 9 or 13, respectively. However, $E_2$ ($10^{-9}\;M$) significantly drove up cell count to 160% of that of control at day 13, while it had a slight but statistically insignificant effect at day 9. $E_2-induced$ stimulation of $[^3H]-thymidine$ incorporation was completely reversed by $E_2$ antagonists (progesterone or tamoxifen). $E_2$ ($10^{-9}\;M$) or EGF ($10^{-8}\;M$) significantly stimulated $[^3H]-thymidine$ incorporation by 144% and 154% of control. $E_2$ plus EGF was synergistic on $[^3H]-thymidine$ incorporation (204% of control), while $E_2$ plus IGF-I showed a slight but no significant synergistic effect. Cell number also displayed similar pattern. $E_2$ ($10^{-9}\;M$) significantly stimulated $P_i$ uptake to 134% of control. $E_2$-induced stimulation of $P_i$ uptake was partially reversed by $E_2$ antagonists. EGF or IGF-I ($10^{-8}\;M$) significantly also increased $P_i$ uptake to 132% or 129% of control. $E_2$ plus EGF had synergistic effect on $P_i$ uptake, while $E_2$ plus IGF-I did not. In conclusion, $E_2$ may act not only directly interaction with its receptors but also indirectly as a modulator of EGF in proliferation and $P_i$ uptake of primary cultured rabbit renal proximal tubular cells.

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Differential Signaling via Tumor Necrosis Factor-Associated Factors (TRAFs) by CD27 and CD40 in Mouse B Cells

  • Woo, So-Youn;Park, Hae-Kyung;Bishop, Gail A.
    • IMMUNE NETWORK
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    • v.4 no.3
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    • pp.143-154
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    • 2004
  • Background: CD27 is recently known as a memory B cell marker and is mainly expressed in activated T cells, some B cell population and NK cells. CD27 is a member of tumor necrosis factor receptor family. Like CD40 molecule, CD27 has (P/S/T/A) X(Q/E)E motif for interacting with TNF receptor-associated factors (TRAFs), and TRAF2 and TRAF5 bindings to CD27 in 293T cells were reported. Methods: To investigate the CD27 signaling effect in B cells, human CD40 extracellular domain containing mouse CD27 cytoplamic domain construct (hCD40-mCD27) was transfected into mouse B cell line CH12.LX and M12.4.1. Results: Through the stimulation of hCD40-mCD27 molecule via anti-human CD40 antibody or CD154 ligation, expression of CD11a, CD23, CD54, CD70 and CD80 were increased and secretion of IgM was induced, which were comparable to the effect of CD40 stimulation. TRAF2 and TRAF3 were recruited into lipid-enriched membrane raft and were bound to CD27 in M12.4.1 cells. CD27 stimulation, however, did not increase TRAF2 or TRAF3 degradation. Conclusion: In contrast to CD40 signaling pathway, TRAF2 and TRAF3 degradation was not observed after CD27 stimulation and it might contribute to prolonged B cell activation through CD27 signaling.

Analysis of Kinematic Motions of First Metatarsophalangeal Joint during Electrical Stimulation of Abductor Hallucis Muscle in Subjects with Hallux Valgus (엄지발가락가쪽휨증의 엄지벌림근 전기자극 시 첫 번째 발허리발가락관절의 운동형상학적 움직임 분석)

  • Kim, Moon-Hwan;Koh, Eun-Kyung;Jung, Do-Young
    • The Journal of Korean Physical Therapy
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    • v.24 no.4
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    • pp.276-281
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    • 2012
  • Purpose: The purpose of this study is to compare the kinematic motion of the first metatarsophalangeal (MTP) joint during an electrical stimulation of abductor hallucis (AbdH) muscle, between the normal group and the hallux valgus (HV) group. Methods: A total of twenty subjects (normal group=10 and HV group=10) participated in this study. The kinematic motions of first MTP joint was measured by using 3-dimensional motion analysis during an electrical stimulation in the sitting position. The intensity of an electrical stimulation was set to be tolerated in each subject, and the data of kinematic motions were collected in three trials of 5 seconds. An independent t-test was used to compare the angle of flexion and abduction of the first MTP joint and proximal phalanx in frontal plane, between the normal and HV groups. Results: Participants showed that the angle of flexion was significantly greater in the HV group ($13.12{\pm}10.61^{\circ}$), compared to that of the normal group ($10.17{\pm}2.31^{\circ}$); and the angle of abduction was significantly smaller in the HV group ($10.61{\pm}4.99^{\circ}$) than that of the normal group. Also, the angle of the proximal phalanx in frontal plane was significantly smaller, compared to the normal group ($53.42{\pm}10.70^{\circ}$) (p<0.05). Conclusion: These findings suggest that dysfunction of AbdH muscle is apparent in HV deformity and provide insight into potential risk factors for the development of HV deformity.

Proinflammatory Effects of High Mobility Group B1 (HMGB1) Versus LPS and the Mechanism of IL-8 Promoter Stimulation by HMGB1 (High mobility group B1(HMGB1)과 LPS의 염증유발효과 차이의 비교 및 HMGB1에 의한 IL-8 promoter 자극 기전의 규명)

  • Jeon, Eun Ju;Kwak, Hee Won;Song, Ju Han;Lee, Young Woo;Chung, Jae Woo;Choi, Jae Chul;Shin, Jong Wook;Park, In Won;Choi, Byoung Whui;Kim, Jae Yeol
    • Tuberculosis and Respiratory Diseases
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    • v.62 no.4
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    • pp.299-307
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    • 2007
  • Background: High mobility group box 1 (HMGB1) is a novel, late mediator of inflammation. This study compared the pro-inflammatory effects of LPS and HMGB1. The transcriptional factors that play an important role in mediating the HMGB1-induced stimulation of IL-8 were also evaluated. Methods: RAW264.7 cells were stimulated with either LPS (100 ng/ml) or HMGB1 (500 ng/ml). The $TNF-{\alpha}$, MIP-2 and $IL-1{\beta}$ levels in the supernatant were evaluated by ELISA at 0, 2, 4, 8, 12 and 24h after stimulation. An acute lung injury was induced by an injection of LPS (5 mg/kg) or HMGB1 (2.5 mg/kg) into the peritoneum of the Balb/c mice. The lung cytokines and MPO activity were measured at 4h (for LPS) or 24h (for HMGB1) after the injection. The transcriptional factor binding sites for NF-IL6, $NF-{\kappa}B$ and AP-1 in the IL-8 promoter region were artificially mutated. Each mutant was ligated with pIL-6luc and transfected into the RAW264.7 cells. One hour after stimulation with HMGB1 (500 ng/ml), the cell lysate was analyzed for the luciferase activity. Results: The expression of MIP-2, which peaked at 8h with LPS stimulation, increased sequentially until 24h after HMGB1 stimulation. An intraperitoneal injection of HMGB1, which induced a minimal increased in $IL-1{\beta}$ expression, provoked the accumulation of neutrophils the lung. A mutation of AP-1 as well as $NF-{\kappa}B$ in the IL-8 promoter region resulted in a lower luciferase activity after HMGB1 stimulation. Conclusion: The proinflammatory effects of HMGB1, particularly on IL-8, are mediated by both $NF-{\kappa}B$ and AP-1.

Dental Caries Risk Can Be Predicted by Simply Measuring the pH and Buffering Capacity of Saliva (치아우식과 연관된 타액의 pH와 완충력)

  • Jeong, Soon-Jeong;Apostolska, Sonja;Jankulovska, Mira;Angelova, Dragana;Nares, Salvador;Yoon, Mi-Sook;Lim, Do-Seon;Angelov, Nikola;Jeong, Moon-Jin
    • Journal of dental hygiene science
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    • v.6 no.3
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    • pp.159-162
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    • 2006
  • This study examined the relationship between the quantity of Streptococcus mutans and Lactobacillus spp. related to dental caries and the degree of acidity in saliva. A total of 240 saliva samples were taken from 80 subjects at the faculty of dentistry in Skopje, Macedonia. The saliva samples were taken by stimulating saliva production stimulation with paraffin chewing. However, no stimulation was applied when obtaining the samples used for measuring the pH. The data showed that in the caries group, S. mutans in 1 ml of saliva formed colonies with confluent growth (CFU > $10^6$ and $10^4-10^5$) in 100% of samples, whereas the Lactobacillus spp. formed colonies with confluent growth in 78.3%. In contrast, no colonies with confluent growth (CFU > $10^6$ and $10^5$) were found in the control group (with healthy intact teeth). In the caries group, the pH of the saliva was slightly acidic (pH = 5.90 - 6.50) and the buffering capacity was very low (below 0.7 ml of saliva per min). On the other hand, the pH of the saliva in the control group was neutral (pH 7.01 - 7.7) and the buffering capacity was high (over 1 ml of saliva per min). The increased number of S. mutans and Lactobacillus spp. in 1 ml of saliva (above $10^5$ CFU or more) from the CRT (Caries Risk Test, Vivadent, Liechtenstein) bacteria test can indicate an increased caries risk as well as slightly higher acidity of the saliva. Overall, these results reveal that the caries risk can be predicted by simply measuring the pH and buffering capacity of saliva, and can be used to monitor the effect of dental hygiene practices with the aim of preventing dental caries.

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Activation of NF-${\kappa}B$ in Lung Cancer Cell Lines in Basal and TNF-${\alpha}$ Stimulated States (폐암 세포에서 기저 상태와 TNF-${\alpha}$ 자극 시 NF-${\kappa}B$의 활성화)

  • HwangBo, Bin;Lee, Seung-Hee;Lee, Choon-Taek;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Kim, Young-Whan
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.5
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    • pp.485-496
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    • 2002
  • Background : The NF-${\kappa}B$ transcription factors control various biological processes including the immune response, acute phase reaction and cell cycle regulation. NF-${\kappa}B$ complexes are retained in the cytoplasm in the basal state and various stimuli cause a translocation of the NF-${\kappa}B$ complexes into the nucleus where they bind to the ${\kappa}B$ elements and regulate the transcription of the target genes. Recent reports also suggest that NF-${\kappa}B$ proteins are involved in oncogenesis, tumor growth and metastasis. High expression of NF-${\kappa}B$ expression was reported in many cancer cell lines and tissues. The constitutive activation of NF-${\kappa}B$ was also reported in several cancer cell lines supporting its role in cancer development and survival. The anti-apoptotic action of NF-${\kappa}B$ is important for cancer survival. NF-${\kappa}B$ also controls the expression of several proteins that are important for cellular adhesion (ICAM-1, VCAM-1) suggesting a role in cancer metastasis. In lung cancer, high expression levels of the NF-${\kappa}B$ subunit p50 and c-Rel were reported. In fact, high expression does not mean a high activity, and the activation pattern of NF-${\kappa}B$ in lung cancer has not been reported. Materials and Methods : In this study, the NF-${\kappa}B$ nuclear binding activity in the basal and TNF-${\alpha}$ stimulated states were exmined in various lung cancer cell lines and compared with the normal bronchial epithelial cell line. Twelve lung cancer cell lines including the non-small cell and small cell lung cancer cell lines (A549, NCI-H358, NCI-H441, NCI-H552, NCI-H2009, NCI-H460, NCI-H1229, NCI-H1703, NCI-H157, NCI-H187, NCI-H417, NCI-H526) and BEAS-2B bronchial epithelial cell line were used. To evaluate the NF-${\kappa}B$ expression and DNA binding activity, western blot analysis and an electrophoretic mobility shift assay with the nuclear protein extracts. Results : The basal expressions of the p65 and p50 subunits were observed in the BEAS-2B cell line and all lung cancer cell lines except for NCI-H358 and NCI-H460. The expression levels of p65 and p50 were increased 30 minutes after stimulation with TNF-${\alpha}$ in BEAS-2B and in 10 lung cancer cell lines. In the NCI-H358 and NCI-H460 cell lines, p65 expression was not observed in the basal and stimulated states and the two p50 related protein levels were higher after stimulation with TNF-${\alpha}$ These new proteins were smaller than p50 and are thought to be variants of p50. In the basal state, NF-${\kappa}B$ was nearly activated in the BEAS-2B and all lung cancer cell lines. The DNA binding activity of the NF-${\kappa}B$ complexes was markedly higher after stimulation with TNF-${\alpha}$ In the BEAS-2B and all lung cancer cell line except for NCI-H358 and NCI-H460, the activated NF-${\kappa}B$ complex was a p65/p50 heterodimer. In the NCI-H358 and NCI-H460 lung cancer cell lines, the NF-${\kappa}B$ complex was variant of a p50/p50 homodimer. Conclusion : The NF-${\kappa}B$ activation pattern in the lung cancer cell lines and the normal bronchial epithelial cell lines was similar except for the activation of a variant of the p50/p50 homodimer in some lung cancer cell linse.

Sono-electrochemical Determination of Uric Acid (요산의 초음파 전기화학적 정량)

  • Cho Hyung-hwa;Bae Zun-ung
    • Journal of the Korean Electrochemical Society
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    • v.3 no.4
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    • pp.232-234
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    • 2000
  • Determination of uric acid by electrochemical method using ultrasonic stimulation has been investigated. Effects of sonication power, sonication time, pH of the solution and temperature were studied to obtain the optimal analytical conditions. The stability of the electrode was also examined. The optimal conditions for the sonovoltammetric determination of uric acid were as follows: temperature, $25.0^{\circ}C$ pH 7.0; sonication power, $20W/cm^2$. The calibration curve for the determination of uric acid by sono-LSV was linear over the range of$8.0{\times}10^{-6}\~5.0\times10^{-4}M$ and the limit of detection was $6.5\times10^{-6}M$.

A high response to controlled ovarian stimulation induces premature luteinization with a negative impact on pregnancy outcomes in a gonadotropin-releasing hormone antagonist cycle

  • Koo, Hwa Seon;Cha, Sun Hwa;Kim, Hye Ok;Song, In Ok;Min, Eung Gi;Yang, Kwang Moon;Park, Chan Woo
    • Clinical and Experimental Reproductive Medicine
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    • v.42 no.4
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    • pp.149-155
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    • 2015
  • Objective: The goal of this study was to investigate the relationship between serum progesterone (P4) levels on the day of human chorionic gonadotropin (hCG) administration and the pregnancy rate among women undergoing controlled ovarian stimulation for in vitro fertilization (IVF) or intracytoplasmic sperm injection-embryo transfer (ICSI-ET) using a flexible antagonist protocol. Methods: This prospective study included 200 IVF and ICSI-ET cycles in which a flexible antagonist protocol was used. The patients were divided into five distinct groups according to their serum P4 levels at the time of hCG administration (0.80, 0.85, 0.90, 0.95, and 1.00 ng/mL). The clinical pregnancy rate (CPR) was calculated for each P4 interval. Statistically significant differences were observed at a serum P4 level of 0.9 ng/mL. These data suggest that a serum P4 concentration of 0.9 ng/mL may represent the optimal threshold level for defining premature luteinization (PL) based on the presence of a significant negative impact on the CPR. Results: The CPR for each round of ET was significantly lower in the PL group defined using this threshold (25.8% vs. 41.8%; p=0.019), and the number of oocytes retrieved was significantly higher than in the non-PL group ($17.3{\pm}7.2$ vs. $11.0{\pm}7.2$; p=0.001). Elevated serum P4 levels on the day of hCG administration were associated with a reduced CPR, despite the retrieval of many oocytes. Conclusion: Measuring serum P4 values at the time of hCG administration is necessary in order to determine the optimal strategy for embryo transfer.

In Vitro Developments of Porcine Embryos Produced by In Vitro Fertilization and Parthenogenetic Activation (체외수정과 단위발생된 돼지 난자의 체외발달 양상의 비교)

  • Uhm, S.J.;Kim, N.H.;Kim, E.Y.;Kim, M.K.;Cha, S.K.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.1
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    • pp.43-49
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    • 1997
  • The objective of this study is to determine developmental pattern and cell allocation to the ICM and TE in haploid and diploid of embryos following parthenogenetic activation and in vitro fertilization. The incidence of development to blastocyst was lower in the combined treatments of ethanol stimulation and cytochalasin B as compared to the control. However, the combined ethanol stimulation and cytochalasin B treatment (diploid) accelerated development to the blastocyst as compared to the ethanol treatment alone (haploid). Significantly reduction in the average number of total cells and ICM was observed in the parthenotes alone as compared to fetilized embryos, but those of combined ethanol stimulation and cytochalasin B treatment embryos were significantly increased as compared to ethanol alone embryos. These results suggested that the ploidy affects preimplantation developmental patters and cell allocation to the ICM and TE in the porcine.

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Cytokines Expression and Nitric Oxide Production under Induced Infection to Salmonella Typhimurium in Chicken Lines Divergently Selected for Cutaneous Hypersensitivity

  • Singh, Rani;Jain, Preeti;Pandey, N.K.;Saxena, V.K.;Saxena, M.;Singh, K.B.;Ahmed, K.A.;Singh, R.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.7
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    • pp.1038-1044
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    • 2012
  • In the present study, the impact of Salmonella Typhimurium on cell-mediated immunity (CMI) was investigated in 5 week-old immuno divergent broiler lines selected for the high and low response to phytohemagglutinin-P. The immune response was assessed in peripheral-blood mononuclear cells (PBMCs) induced with Salmonella Typhimurium at different time intervals (0 h, 0.5 h, 2 h, 4 h, 6 h, 12 h and 24 h). The differential mRNA expression patterns of IFN-${\gamma}$, IL-2 and iNOS were evaluated by quantitative real time PCR. In-vitro production of nitric oxide (NO) was also estimated in the culture supernatant and correlated with iNOS mRNA expression. Present study showed higher production of NO in the high cell-mediated line (HCMI) as compared to the low cell-mediated line (LCMI) upon stimulation with Salmonella Typhimurium. Correspondingly, higher mRNA expression of iNOS and IFN-${\gamma}$ were observed in high response birds (HCMI); but IL-2 was down regulated in this line compared to the low response birds (LCMI). Significantly (p<0.05) higher expression of iNOS, IFN-${\gamma}$ and higher production of NO in high line indicated that the selection for PHA-P response might be employed for increasing the immune competence against Salmonella Typhimurium in chicken flocks.