• Title/Summary/Keyword: G Proteins

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Immune reactions between excretory-secretory antigens and specific antibodies of Clonorchis sinensis before and after praziquantel treatment in experimentally infected rabbits (간흡충 감염 토끼에서 프라지콴텔 치료 전후의 특이항체의 간흡충 분비배설항원에 대한 면역반응양상)

  • 김석일
    • Parasites, Hosts and Diseases
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    • v.32 no.1
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    • pp.35-42
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    • 1994
  • This study was designed to evaluate the humoral immune reactions in clonorchiasis before and after praziquantel treatment. Rabbits were infected with 150 or 450 metacercariae, treated on 4 and 8111 months after infection, and observed for 13 months of posttreatment. Infection controls were maintained for 22 months. Antigen was the metabolic product of worms incubated in physiologic saline. The immune reactions of anti-clonorchis IgG were observed using SDS-PAGE/immunoblot. During the Infection and Posttreatment, the antigenic Proteins of 66, 63, 54, 52, 50,47,42, 40, 38, 34,33,30, 27, 25, 23, 20, 19, 18, 17, 16, 15, 14, 13, 12.5 and 11.5 kDa were detected. Of them, 33,27, 13, and 12.5-kDa antigens were highly antigenic and observed predominently in infection controls. After the treatment, 13 and 12.5-kDa antigens faded in 6 months after the second treatment, but 33 and 27-kDa antigens were detected until 13 months of posttreatment. The results clearly demonstrate that 13 and 12.5-kDa antigens represent attenuated host immune reactions by praziquantel treatment. As the 12.5-kDa antigen had a large amount of protein in SDS-PAGE, it was designated as'K2-Ag'of C. sinenis.

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A New Histone Deacetylase Inhibitor, MHY4381, Induces Apoptosis via Generation of Reactive Oxygen Species in Human Prostate Cancer Cells

  • Richa, Sachan;Dey, Prasanta;Park, Chaeun;Yang, Jungho;Son, Ji Yeon;Park, Jae Hyeon;Lee, Su Hyun;Ahn, Mee-Young;Kim, In Su;Moon, Hyung Ryong;Kim, Hyung Sik
    • Biomolecules & Therapeutics
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    • v.28 no.2
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    • pp.184-194
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    • 2020
  • Histone deacetylase (HDAC) inhibitors represent a novel class of anticancer agents, which can be used to inhibit cell proliferation and induce apoptosis in several types of cancer cells. In this study, we investigated the anticancer activity of MHY4381, a newly synthesized HDAC inhibitor, against human prostate cancer cell lines and compared its efficacy with that of suberoylanilide hydroxamic acid (SAHA), a well-known HDAC inhibitor. We assessed cell viability, apoptosis, cell cycle regulation, and other biological effects in the prostate cancer cells. We also evaluated a possible mechanism of MHY4381 on the apoptotic cell death pathway. The IC50 value of MHY4381 was lower in DU145 cells (IC50=0.31 µM) than in LNCaP (IC50=0.85 µM) and PC-3 cells (IC50=5.23 µM). In addition, the IC50 values of MHY4381 measured in this assay were significantly lower than those of SAHA against prostate cancer cell lines. MHY4381 increased the levels of acetylated histones H3 and H4 and reduced the expression of HDAC proteins in the prostate cancer cell lines. MHY4381 increased G2/M phase arrest in DU145 cells, and G1 arrest in LNCaP cells. It also activated reactive oxygen species (ROS) generation, which induced apoptosis in the DU145 and LNCaP cells by increasing the ratio of Bax/Bcl-2 and releasing cytochrome c into the cytoplasm. Our results indicated that MHY4381 preferentially results in antitumor effects in DU145 and LNCaP cells via mitochondria-mediated apoptosis and ROS-facilitated cell death pathway, and therefore can be used as a promising prostate cancer therapeutic.

Effect of Sphingosine-1-Phosphate on Intracellular Free Ca2+ in Cat Esophageal Smooth Muscle Cells

  • Lee, Dong Kyu;Min, Young Sil;Yoo, Seong Su;Shim, Hyun Sub;Park, Sun Young;Sohn, Uy Dong
    • Biomolecules & Therapeutics
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    • v.26 no.6
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    • pp.546-552
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    • 2018
  • A comprehensive collection of proteins senses local changes in intracellular $Ca^{2+}$ concentrations ($[Ca^{2+}]_i$) and transduces these signals into responses to agonists. In the present study, we examined the effect of sphingosine-1-phosphate (S1P) on modulation of intracellular $Ca^{2+}$ concentrations in cat esophageal smooth muscle cells. To measure $[Ca^{2+}]_i$ levels in cat esophageal smooth muscle cells, we used a fluorescence microscopy with the Fura-2 loading method. S1P produced a concentration-dependent increase in $[Ca^{2+}]_i$ in the cells. Pretreatment with EGTA, an extracellular $Ca^{2+}$ chelator, decreased the S1P-induced increase in $[Ca^{2+}]_i$, and an L-type $Ca^{2+}$-channel blocker, nimodipine, decreased the effect of S1P. This indicates that $Ca^{2+}$ influx may be required for muscle contraction by S1P. When stimulated with thapsigargin, an intracellular calcium chelator, or 2-Aminoethoxydiphenyl borate (2-APB), an $InsP_3$ receptor blocker, the S1P-evoked increase in $[Ca^{2+}]_i$ was significantly decreased. Treatment with pertussis toxin (PTX), an inhibitor of $G_i$-protein, suppressed the increase in $[Ca^{2+}]_i$ evoked by S1P. These results suggest that the S1P-induced increase in $[Ca^{2+}]_i$ in cat esophageal smooth muscle cells occurs upon the activation of phospholipase C and subsequent release of $Ca^{2+}$ from the $InsP_3$-sensitive $Ca^{2+}$ pool in the sarcoplasmic reticulum. These results suggest that S1P utilized extracellular $Ca^{2+}$ via the L type $Ca^{2+}$ channel, which was dependent on activation of the $S1P_4$ receptor coupled to PTX-sensitive $G_i$ protein, via phospholipase C-mediated $Ca^{2+}$ release from the $InsP_3$-sensitive $Ca^{2+}$ pool in cat esophageal smooth muscle cells.

The Roles of Immune Regulatory Factors FoxP3, PD-1, and CTLA-4 in Chronic Viral Infection (만성 바이러스 감염에서 면역조절인자 FoxP3, PD-1 및 CTLA-4의 역할)

  • Cho, Hyosun
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.221-227
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    • 2013
  • Human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV) cause viral infections that lead to chronic diseases. When they invade human body, virus specific T cells play an important role in antiviral effector functions including killing virus-infected cells and helping B cells to produce specific antibodies against viral proteins. The antiviral activity of T cells is usually affected by immune-regulatory factors that express on surface of T cells. Recently, many researchers have investigated the relationship between effector functions of virus specific T cells and characteristics of immune regulatory factors (e.g., CD28, CD25, CD45RO, FoxP3, PD-1, CTLA-4). In particular, Immune inhibitory molecules such as forkhead box P3 (FoxP3), programmed death-1 (PD-1), and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) are associated with T-cell dysfunction. They are shown to be up-regulated in chronic viral diseases such as hepatitis B, hepatitis C or human immunodeficiency virus infection. Therefore, the positive correlation between viral persistence and expression of immune regulatory factors (FoxP3, PD-1, and CTLA-4) has been suggested. In this review, the roles of immune regulatory factors FoxP3, PD-1, and CTLA-4 were discussed in chronic viral diseases such as HIV, HBV, or HCV.

Studies on Changes of Isoflavone and Nutrients during Germination of Soybean Varieties (발아기간에 따른 대두 품종별 이소플라본 및 영양성분 변화)

  • Jeon, Su-Hyeon;Lee, Kyong-Ae;Byoun, Kwang-Eui
    • Korean Journal of Human Ecology
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    • v.14 no.3
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    • pp.485-489
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    • 2005
  • The purpose of this study is to investigate changes of isoflavone and nutrients during germination of soybean varieties and to determine the optimal germination condition for soybean processing. After 5 days' germination, water content of the soybeans increased in the order of Poongsannamoolkong> Sinpaldal#2> Black#1> Jinpumkong. Ash content was higher in the order of Poongsannamoolkong> Sinpaldal#2> Jinpumkong> black#1 soybean. All varieties of the soybean samples increased in ash content during germination. Crude fat content was higher in the order of Poongsannamoolkong> Black#1> Jinpumkong> Shinpaldalkong. Crude protein content was higher in the order of Shinpaldalkong> Black#1> Jinpumkong> Poongsannamoolkong. Polypeptide subunits separated from the SDS-PAGE of the entire proteins were showing comparatively high level in the range of 2-16KD in Jinpumkong and 2-21KD in Poongsannamoolkong till the first day of germination, 2-25KD in Sinpaldal#2 till the second day, and 6-27KD in Black#l till the third day. Changes of total isoflavone contents during germination showed different patterns in soybean varieties: Jinpumkong and Black#1 had the highest total isoflavone content on the fourth day of germination, Poongsannamoolkong and Sinpaldal#2 on the first and the second day of germination, respectively.

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Action and Signaling of Lysophosphatidylethanolamine in MDA-MB-231 Breast Cancer Cells

  • Park, Soo-Jin;Lee, Kyoung-Pil;Im, Dong-Soon
    • Biomolecules & Therapeutics
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    • v.22 no.2
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    • pp.129-135
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    • 2014
  • Previously, we reported that lysophosphatidylethanolamine (LPE), a lyso-type metabolite of phosphatidylethanolamine, can increase intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) via type 1 lysophosphatidic acid (LPA) receptor ($LPA_1$) and CD97, an adhesion G-protein-coupled receptor (GPCR), in MDA-MB-231 breast cancer cells. Furthermore, LPE signaling was suggested as like $LPA_1/CD97-G_{i/o}$ proteins-phospholipase $C-IP_3-Ca^{2+}$ increase in these cells. In the present study, we further investigated actions of LPE not only in the $[Ca^{2+}]_i$ increasing effect but also in cell proliferation and migration in MDA-MB-231 breast cancer cells. We utilized chemically different LPEs and a specific inhibitor of $LPA_1$, AM-095 in comparison with responses in SK-OV3 ovarian cancer cells. It was found that LPE-induced $Ca^{2+}$ response in MDA-MB-231 cells was evoked in a different manner to that in SK-OV3 cells in terms of structural requirements. AM-095 inhibited LPE-induced $Ca^{2+}$ response and cell proliferation in MDA-MB-231 cells, but not in SK-OV3 cells, supporting $LPA_1$ involvement only in MDA-MB-231 cells. LPA had significant effects on cell proliferation and migration in MDA-MB-231 cells, whereas LPE had less or no significant effect. However, LPE modulations of MAPKs (ERK1/2, JNK and p38 MAPK) was not different to those by LPA in the cells. These data support the involvement of LPA1 in LPE-induced $Ca^{2+}$ response and cell proliferation in breast MDA-MB-231 cells but unknown GPCRs (not $LPA_1$) in LPE-induced responses in SK-OV3 cells. Furthermore, although LPE and LPA utilized $LPA_1$, LPA utilized more signaling cascades than LPE, resulting in stronger responses by LPA in proliferation and migration than LPE in MDA-MB-231 cells.

Effects of Water-Extracts of Bambusae concretio silicae(BCS, 天竺黃) on the Expression of Neurotransmitter Receptors (천죽황(Bambusae concretio silicae, 天竺黃) 물추출물이 신경전달물질 수용체의 표현에 미치는 영향)

  • Yoon, Jae-Hong;Kim, Kyung-Min;Kim, Kyung-Hun;Shin, Gil-Cho;Jeong, Seung-Hyun
    • Journal of Oriental Neuropsychiatry
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    • v.22 no.2
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    • pp.163-176
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    • 2011
  • Objectives : BCS(Bambusae concretio silicae) is used as a traditional medicine in Korea for the incipient stroke. Recent reports indicated that BCS has a neuroprotective effect by anti-convulsion effect. However, it's mechanism is not well studied. The purpose of this study was to investigate into the molecular mechanism of BCS for neuroprotection in normoxia of cultured rat hippocampal neurons. Methods : BCS (1.0, 2.5, 5.0, and $10.0\;{\mu}g/m{\ell}$) was added to culture media (Neurobasal supplemented with B27) on DIV 0, given a normoxia, and the cell viability was measured by typical phase-contrast images of the cultures with 1.0, 2.5, 5.0, and $10.0\;{\mu}g/m{\ell}$ on DIV 21. Effects of BCS on the expression of various synaptic proteins ($GABA_B$ R1, $GABA_B$ R2, GlyR, PSD95) were observed by immunocytochemistry showing on DIV 3, 7 and 21. Results : Typical phase-contrast images of the cultures indicated that BCS has a protective effect of rat hippocampal cells in normoxia. The BCS upregulated $GABA_B$ R1 after normoxia on DIV 7, $GABA_A$ ${\beta}2/3$ on DIV 21 and $GABA_B$ R2 on DIV 21. And the BCS downregulated PSD95 after normoxia on DIV 7. Conclusions : The present study showed evidence for the efficacy of BCS in Typical phase-contrast images, upregulation of inhibitory neurotransmitter receptors($GABA_B$ R1) and downregulation of PSD95 which eventually protected neuronal cell death in normoxia.

Effect on the Production of Toxin Protein of Bacillus thuringiesis by Culture Conditions (배양 조건이 Bacillus thuringiesis 독소단백질의 생산에 미치는 영향)

  • Kim, Tae Wan;Kim, Tae Hwan;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.358-363
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    • 2014
  • We was investigated parasporal inclusion proteins change to use industrial medium of new strain Bacillus thuringiensis CAB 565, CAB 566. To confirm medium's oxygen efficient consist of glucose and yeast extract, we was conducted oxygen transfer coefficients (KLa) of medium's concentration and impeller in 20 l-Jar fermentor. When to increase air flow rate and medium concentration, KLa rate is rise. Also it is effective on agitation rate 200 rpm, but KLa rate is decrease when to rise agitation rate. To hold dissolved oxygen rate (upper 50%), Air flow rate is steadily increase on culture to use microsparger. When 16 hour of culture stage, B.t. CAB 565 and B.t. CAB 566 harvested respectively $2.3{\times}10^{10}$, $1.8{\times}10^{10}$ viable cell/ml. When 54 hour, B.t. CAB565, 566 harvested respectively $1.9{\times}10^{10}$, $1.4{\times}10^{10}spore/ml$. To resulting carbon's concentration, It is the most effective that glucose concentration is contained 5% in medium.

Identifying Genes Related with Radiation Resistance in Oral Squamous Cell Carcinoma Cell Lines (두경부 편평상피세포암주에서 방사선 내성 관련 유전인자의 확인)

  • Lee, Sei-Young;Seol, Jeong-Hun;Park, Haeng-Ran;Cho, Nam-Hoon;Choi, Yoon-Pyo;Rha, Sun-Young;Baek, Seung-Jae;Hwang, Jun-Yeon;Kim, Se-Heon
    • Korean Journal of Head & Neck Oncology
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    • v.27 no.2
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    • pp.190-197
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    • 2011
  • Background and Objective : Radiation resistance(RR) is one of main determinants of treatment outcome in oral squamous cell carcinoma(OSCC), but accurate prediction of RR is difficult. We aim to establish RR OSCC cell lines and identify genes related with RR by a measurement of altered gene expression after inducing RR. Material and Methods : OSCC cell lines, SCC15, SCC25 and QLL1, were treated with 2Gy radiation per session, and parts of them were alive in finally accumulated dosage of 60Gy through 30 times repetition of radiotherapy for inducing RR cell lines. We compared results of cDNA array and proteomics in non-radiated cell lines and RR cell lines to detect changes of gene expression. Western blot was used for the validation of results. Results : cDNA array revealed 265 commonly up-regulated genes and 268 commonly down-regulated genes in 3 RR cell lines comparing their non-radiated counterpart. Among them, 30 cancer related genes were obtained. Proteomics showed 51 commonly altered protein expressions in 3 RR cell lines and 18 cancer related proteins were obtained. Among the detected genes, we found NM23-H1 and PA2G4 were over-expressed in both cDNA array and proteomics. Western blot showed increased expression of NME1 in RR cell lines but not in PA2G4. Conclusion: We concluded that NM23-H1 may be a candidate of RR related gene and over-expression of NM23-H1 could be a biomarker to predict RR in OSCC.

The Change in the Amounts of Proteins and Lipids Deposited on Soft Contact Lens Caused by Drinking (음주로 인한 소프트콘택트렌즈의 단백질 및 지질 침착양 변화)

  • Kim, So Ra;Lim, Shin Gyu;Bae, Seok Chun;Choi, Jung Hyun;Park, Sang Hee;Park, Mijung
    • Journal of Korean Ophthalmic Optics Society
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    • v.17 no.2
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    • pp.233-239
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    • 2012
  • Purpose: In the study, the change of protein and lipid deposits on soft contact lens by drinking was investigated. Methods: Fifty male subjects wearing soft contact lens were surveyed whether they felt any discomfort induced by drinking or not. Further, 32 male subjects who has no ocular disease drank 190 mL alcohol. The protein and lipid deposits on soft contact lens (etafilcon A material) of subjects were measured after 4 hours later and compared with those of non-drinking subject. Results: When subjects drink alcohol with soft contact lens on, 58% of subjects answered they experienced the change of lens awareness such as stiffness, blurry sight, dryness and so on. The protein deposit on soft contact lens increased an average of $59.3{\mu}g/lens$ by drinking and the case of more than double in protein deposit was reached in 9 eyes. However, the protein deposited on soft contact lens was lysozyme which was unchanged by drinking. The amounts of cholesterol and methyl oleate after drinking were 85.5% (p=0.25) and 52.6% (p=0.002) of non-drinking's indicating some change of lipid deposit on soft contact lens by drinking. Conclusions: The results showed the composition of protein and lipid deposited on soft contact lens was changed due to drinking. Thus, it is suggested that wearing soft contact lens when drinking might be one of the reasons to feel discomfort.