• Title/Summary/Keyword: biochemical pathway

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Effect of Ketoconazole, a Cytochrome P450 Inhibitor, on the Efficacy of Quinine and Halofantrine against Schistosoma mansoni in Mice

  • Seif el-Din, Sayed Hassan;Abdel-Aal Sabra, Abdel-Nasser;Hammam, Olfat Ali;El-Lakkany, Naglaa Mohamed
    • Parasites, Hosts and Diseases
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    • v.51 no.2
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    • pp.165-175
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    • 2013
  • The fear that schistosomes will become resistant to praziquantel (PZQ) motivates the search for alternatives to treat schistosomiasis. The antimalarials quinine (QN) and halofantrine (HF) possess moderate antischistosomal properties. The major metabolic pathway of QN and HF is through cytochrome P450 (CYP) 3A4. Accordingly, this study investigates the effects of CYP3A4 inhibitor, ketoconazole (KTZ), on the antischistosomal potential of these quinolines against Schistosoma mansoni infection by evaluating parasitological, histopathological, and biochemical parameters. Mice were classified into 7 groups: uninfected untreated (I), infected untreated (II), infected treated orally with PZQ (1,000 mg/kg) (III), QN (400 mg/kg) (IV), KTZ (10 mg/kg)+QN as group IV (V), HF (400 mg/kg) (VI), and KTZ (as group V)+HF (as group VI) (VII). KTZ plus QN or HF produced more inhibition (P<0.05) in hepatic CYP450 (85.7% and 83.8%) and CYT b5 (75.5% and 73.5%) activities, respectively, than in groups treated with QN or HF alone. This was accompanied with more reduction in female (89.0% and 79.3%), total worms (81.4% and 70.3%), and eggs burden (hepatic; 83.8%, 66.0% and intestinal; 68%, 64.5%), respectively, and encountering the granulomatous reaction to parasite eggs trapped in the liver. QN and HF significantly (P<0.05) elevated malondialdehyde levels when used alone or with KTZ. Meanwhile, KTZ plus QN or HF restored serum levels of ALT, albumin, and reduced hepatic glutathione (KTZ+HF) to their control values. KTZ enhanced the therapeutic antischistosomal potential of QN and HF over each drug alone. Moreover, the effect of KTZ+QN was more evident than KTZ+HF.

Effect of Dietary Folate on Hyperhomocysteinemia and Cellular Toxicity Induced Alcohol Administration in Rat Liver

  • Kim, Chung-Hyeon;Kim, Ki-Nam;Kim, Yeon-Soo;Chang, Nam-Soo
    • Molecular & Cellular Toxicology
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    • v.1 no.2
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    • pp.137-141
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    • 2005
  • The critical role of folate in the remethylation pathway for methionine synthesis from homocysteine has been well documented. Hyperhomocysteinemia resulting from inadequate folate nutrition has been implicated in increased incidence of macrovascular diseases, colorectal cancer, neural tube defects, etc. Chronic exposure to ethanol impairs folate nutrition and one-carbon metabolism in the liver, which often results in fatty liver due to a defective remetylation process. This study was carried out to investigate the chronic effects of moderate levels of alcohol and dietary folate on plasma homocysteine levels, and on histopathology and biochemical functions of the liver. Rats were raised on experimental diets with three levels of folate (0, 2, 8 mg/kg diet), and 50% ethanol (1.8 ml/kg body weight) was administered intragastically by intubation tubes three times a week for 10 weeks. Plasma homocysteine concentrations were found to be significantly influenced by dietary folate intake and alcohol administration. Among all treatment groups, plasma homocysteine levels were the highest in the animals receiving a combined treatment of folate deficient diet and alcohol administration. Plasma homocysteine concentrations were negatively correlated with folate concentration in the plasma (p<0.01) and liver (p<0.05). Among alcohol treated rats, increase in plasma homocysteine values due to macrovascular and microvascular fatty changes and spotted necrosis were observed more frequently in folate-deficient animals diet than those on folate-adequate and folate supplemented diets in alcohol-treated rats. These results indicate that folate supplementation above the recommended level might be beneficial in the prevention of alcohol-related hyperhomocysteinemia and abnormal histologic changes in the liver.

Cell Death Induction Mechanism of Non-small Cell Lung Cancer Cell Line, NCI-H1703 by Docetaxel (Docetaxel에 의한 비소세포폐암세포주 NCI-H1703의 세포사멸 유도기전)

  • Ha Hyeon-Cheol;Song Seung-Hwan;Park Chin-Su;Kim Jong-Won;Kim Yeong-Dae
    • Journal of Chest Surgery
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    • v.39 no.9 s.266
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    • pp.668-673
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    • 2006
  • Background: Docetaxel has been effectively used as an anti-cancer chemotherapuetic agent for various tumor treatments including lung cancer. However, the cell death induction mechanism(s) involved with docetaxel treatment in lung cancer cells has not been known yet. Material and Method: In the present study, the cellular and biochemical changes of NCI-H1703 cells (non-small cell lung cancer cell line, p53-mutant) after docetaxel treatment have been monitored by flow cytometry, fluorescence microscopy and western blot. Result: Docetaxel treatment significantly resulted in decrease of S phase as well as increase of G2 phase, and consequently evoked an increase of cell death in NCI-H1703 cells. After docetaxel exposure the activations of caspase-3 and caspase-9 were detected. Conclusion: Take together, it is suggested that the docetaxel induces NCI-H1703 cell death by caspase-9 and caspase-3 dependent mitochondrial apoptotic pathway.

DIFFERENTIATION OF ADULT STEM CELL DERIVED FROM BUCCAL FAT PAD INTO OSTEOBLAST (협부지방에서 성체 줄기세포의 분리와 골모 세포로의 분화)

  • Pyo, Sung-Woon;Park, Jang-Woo;Lee, Il-Kyu;Kim, Chang-Hyen
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.6
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    • pp.524-529
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    • 2006
  • For the repairing of bone defect, autogenous or allogenic bone grafting remains the standard. However, these methods have numerous disadvantages including limited amount, donor site morbidity and spread of diseases. Tissue engineering technique by culturing stem cells may allow for a smart solution for this problem. Adipose tissue contains mesenchymal stem cells that can be differentiate into bone, cartilage, fat or muscle by exposing them to specific growth conditions. In this study, the authors procured the stem cell from buccal fat pad and differentiate them into osteoblast and are to examine the bone induction capacity. Buccal fat-derived cells (BFDC) were obtained from human buccal fat pad and cultured. BFDC were analyzed for presence of stem cell by immunofluorescent staining against CD-34, CD-105 and STRO-1. After BFDC were differentiated in osteogenic medium for three passages, their ability to differentiate into osteogenic pathway were checked by alkaline phosphatase (ALP) staining, Alizarin red staining and RT-PCR for osteocalcin (OC) gene expression. Immunofluorescent and biochemical assays demonstrated that BFDC might be a distinguished stem cells and mineralization was accompanied by increased activity or expression of ALP and OC. And calcium phosphate deposition was also detected in their extracelluar matrix. The current study supports the presence of stem cells within the buccal fat pad and the potential implications for human bone tissue engineering for maxillofacial reconstruction.

Mode of Action on EcoRI Restriction Endonuclease: EcoRI and EcoRI Variant N199H have Active Monomeric Forms

  • Kim, Jae-Jong;Koh, Suk-Hoon;Kim, Joong-Su;Lee, Dae-Sil
    • BMB Reports
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    • v.31 no.2
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    • pp.149-155
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    • 1998
  • The N199H variant of the EcoRI endonuclease has about twice the catalytic activity of the wild-type. A comparison of their biochemical characteristics, using synthetic oligonucleotides 5'-dAAAACTTAAGAAAAAAAAAAA-3' (KA) and 5'-dTTTTTGAATTCTTTTTTTTTT-3' (KT), helps to define the cleavage reaction pathway of these enzymes. Both EcoRI and EcoRI variant N199H were found to cleave singlestranded KA or KT about three times faster than the double-stranded forms, although the KT oligonucleotide was more susceptible. Using the ssDNA substrate in kinetic analyses, lower $K_m$ values were obtained for the N199H variant than for the wild-type at low (50 mM), as well as high (200 mM), sodium chloride concentrations. This difference between the endonucleases is attributed to a grealter accessibility for tbe substrate by the variant, and also a higher affinity for the DNA backbone. It also appears that the relative activities of the two enzymes, particularly at high ionic strength, are proportional to their populations in the monomeric enzyme form. That is, according to gel filtration data, half of the N199H molecules exist as monomers in 200 mM NaCl, whereas those of the wild-type are mainly dimeric. Consequently, the Asp199 residue of the EcoRI endonuclease may be implicated in the protein-protein interaction leading to dimerization, as well as in coupling to DNA substrates. In summary, it is proposed that active monomeric endonuclease molecules, derived from the dimeric enzyme, recognize and form a complex with a single stranded form of the DNA substrate, which then undergoes nucleophilic substitution and cleavage.

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Effect of cold stress on infanticide by female Swiss albino mice Mus musculus: a pilot study

  • Zafar, Tabassum;Naik, Ab Qayoom;Shrivastava, Vinoy K.
    • Journal of Animal Science and Technology
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    • v.60 no.4
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    • pp.7.1-7.5
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    • 2018
  • Background: Mice are widely accepted research models of great clinical significance. Maintenance of laboratory mice breed is an essential aspect for performing research activities in various fields of science. Infanticide is one of the prominent causes of litter loss during maintenance of laboratory mice stock. The present study is an effort to monitor the effect of change in ambient temperature of female mice below the normal range on cannibalism and infanticide during early postparturition phase. Adult female Swiss albino mice have been divided into two groups of control and treatment. On the day of litter group one was maintained under controlled temperature conditions (minimum $20^{\circ}C$ to maximum $23^{\circ}C$) throughout, while female mice belong to group two have been exposed to variation of room temperature (maximum $15^{\circ}C$ to minimum $10^{\circ}C$ for two nights and one day) until 36 h postparturition. Results: The effects of temperature changes were observed on the infanticide behaviour of dams along with the survival of pups in early postparturition phase till 36 h after delivery. The significant statistical difference (P < 0.05) was reported in infanticide behaviour of dams when control and treatment group was compared. It is observed that decrement in surrounding temperature promotes decrement in the ambient body temperature of dams during early postparturition. It is proposed that alteration of hypothalamic homeostasis due to temperature change induces cannibalism and infanticide behaviour. Lack of thermoregulation during early postparturition creates the sense of insecurity, in-satiety, anxiety and stress. Conclusions: Authors strongly recommend the maintenance of body and surrounding temperature to prevent infanticidal behaviour and cannibalism within Swiss albino mice population. Further investigations are advisable to authenticate the active behavioural and biochemical pathway behind the phenomena.

Potential for Dependence on Lisdexamfetamine - In vivo and In vitro Aspects

  • Yun, Jaesuk;Lee, Kwang-Wook;Eom, Jang-Hyeon;Kim, Young-Hoon;Shin, Jisoon;Han, Kyoungmoon;Park, Hye-Kyung;Kim, Hyung Soo;Cha, Hye Jin
    • Biomolecules & Therapeutics
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    • v.25 no.6
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    • pp.659-664
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    • 2017
  • Although lisdexamfetamine is used as a recreational drug, little research exists regarding its potential for dependence or its precise mechanisms of action. This study aims to evaluate the psychoactivity and dependence profile of lisdexamfetamine using conditioned place preference and self-administration paradigms in rodents. Additionally, biochemical techniques are used to assess alterations in the dopamine levels in striatal synaptosomes following administration of lisdexamfetamine. Lisdexamfetamine increased both conditioned place preference and self-administration. Moreover, after administration of the lisdexamfetamine, dopamine levels in the striatal synaptosomes were significantly increased. Although some modifications should be made to the analytical methods, performing high performance liquid chromatography studies on synaptosomes can aid in predicting dependence liability when studying new psychoactive substances in the future. Collectively, lisdexamfetamine has potential for dependence possible via dopaminergic pathway.

Isolation of Pseudomonas putida Z104 and Degra-dation Characteristics of Benzoate and Catechol (Benzoate와 Catechol을 분해하는 Pseudomonas putida Z104의 분리 및 분해특성)

  • 김기필;김준호;김민옥;박정아;정원화;김치경
    • Korean Journal of Environmental Biology
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    • v.18 no.3
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    • pp.307-313
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    • 2000
  • Aromatic hydrocarbons are known to be recalcitrant, so that they have been concerned as pollutant chemicals. Microorganisms play a major role in the breakdown and mineralization of these compounds. However, the kinetics of the biodegradation process may be much slower than desired from environmental consideration. The biodegradation of aromatic hydrocarbons is conducted by oxidation to produce catechol as a common intermediate which is metabolized for carbon and energy sources. In this study, a bacterial isolate capable of degrading several aromatic hydrocarbons was isolated from the contaminated wastewater of Yeocheon industrial complex. On the basis of biochemical characteristics and major cellular fatty acids, the isolate was identified as Pseudomonas putida Z104. The strain Z104 can utilize benzoate and catechol as the sole carbon and energy sources via a serial degradative pathway. The strain degraded actively 0.5 mM catechol in MM2 medium at pH 7.0 and 3$0^{\circ}C$.

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Imperatorin Suppresses Degranulation and Eicosanoid Generation in Activated Bone Marrow-Derived Mast Cells

  • Jeong, Kyu-Tae;Lee, Eujin;Park, Na-Young;Kim, Sun-Gun;Park, Hyo-Hyun;Lee, Jiean;Lee, Youn Ju;Lee, Eunkyung
    • Biomolecules & Therapeutics
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    • v.23 no.5
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    • pp.421-427
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    • 2015
  • Imperatorin has been known to exert many biological functions including anti-inflammatory activity. In this study, we investigated the inhibitory effects of imperatorin on the production of inflammatory mediators in mouse bone marrow-derived mast cells (BMMC). Imperatorin inhibited degranulation and the generation of eicosanoids (leukotriene $C_4$ ($LTC_4$) and prostaglandin $D_2$ ($PGD_2$) in IgE/antigen (Ag)-stimulated BMMC. To elucidate the molecular mechanism involved in this process, we investigated the effect of imperatorin on intracellular signaling in BMMC. Biochemical analyses of the IgE/Ag-mediated signaling pathway demonstrated that imperatorin dramatically attenuated degranulation and the production of 5-lipoxygenase-dependent $LTC_4$ and cyclooxygenase-2-dependent $PGD_2$ through the inhibition of intracellular calcium influx/phospholipase $C{\gamma}1$, cytosolic phospholipase $A_2$/mitogen-activated protein kinases and/or nuclear factor-${\kappa}B$ pathways in BMMC. These results suggest that the effects of imperatorin on inhibition of degranulation and eicosanoid generation through the suppression of multiple steps of IgE/Ag-mediated signaling pathways would be beneficial for the prevention of allergic inflammation.

Cripto Enhances Proliferation and Survival of Mesenchymal Stem Cells by Up-Regulating JAK2/STAT3 Pathway in a GRP78-Dependent Manner

  • Yun, SeungPil;Yun, Chul Won;Lee, Jun Hee;Kim, SangMin;Lee, Sang Hun
    • Biomolecules & Therapeutics
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    • v.26 no.5
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    • pp.464-473
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    • 2018
  • Cripto is a small glycosylphosphatidylinositol-anchored signaling protein that can detach from the anchored membrane and stimulate proliferation, migration, differentiation, vascularization, and angiogenesis. In the present study, we demonstrated that Cripto positively affected proliferation and survival of mesenchymal stem cells (MSCs) without affecting multipotency. Cripto also increased expression of phosphorylated janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), 78 kDa glucose-regulated protein (GRP78), c-Myc, and cyclin D1. Notably, treatment with an anti-GRP78 antibody blocked these effects. In addition, pretreatment with STAT3 short interfering RNA (siRNA) inhibited the increase in p-JAK2, c-Myc, cyclin D1, and BCL3 levels caused by Cripto and attenuated the pro-survival action of Cripto on MSCs. We also found that incubation with Cripto protected MSCs from apoptosis caused by hypoxia or $H_2O_2$ exposure, and the level of caspase-3 decreased by the Cripto-induced expression of B-cell lymphoma 3-encoded protein (BCL3). These effects were sensitive to down-regulation of BCL3 expression by BCL3 siRNA. Finally, we showed that Cripto enhanced expression levels of vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), and hepatocyte growth factor (HGF). In summary, our results demonstrated that Cripto activated a novel biochemical cascade that potentiated MSC proliferation and survival. This cascade relied on phosphorylation of JAK2 and STAT3 and was regulated by GRP78. Our findings may facilitate clinical applications of MSCs, as these cells may benefit from positive effects of Cripto on their survival and biological properties.