• Title/Summary/Keyword: Detoxification.

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The Beneficial Effects of Ferulic Acid supplementation during In Vitro Maturation of Porcine Oocytes on Their Parthenogenetic Development

  • Lee, Kyung-Mi;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.32 no.4
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    • pp.257-265
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    • 2017
  • Ferulic Acid (FA) is a metabolite of phenylalanine and tyrosine, a phenolic compound commonly found in fruits and vegetables. Several studies have shown that FA has various functions such as antioxidant effect, prevention of cell damage from irradiation, protection from cell damage caused by oxygen deficiency, anti-inflammatory action, anti-aging action, liver protective effect and anti-cancer action. In this study, we investigated the maturation rate, intracellular glutathione (GSH) and reactive oxygen species (ROS) of porcine oocytes by adding FA to the in vitro maturation (IVM) medium and examined subsequent embryonic developmental competence at 5% oxygen through parthenogenesis. There is no significant difference between the control group ($0{\mu}M$) and treatment groups ($5{\mu}M$, $10{\mu}M$, $20{\mu}M$) on maturation rates. Intracellular GSH levels in oocyte treated with $5{\mu}M$ of FA significantly increased (P < 0.05), and $20{\mu}M$ of FA revealed significant decrease (P < 0.05) in intracellular ROS levels compared with the control group. Oocytes treated with FA exhibited significantly higher cleavage rates (79.01% vs 89.19%, 92.20%, 90.89%, respectively) than the control group. Oocytes treated with $10{\mu}M$ showed significantly higher blastocyst formation rates (28.3% vs 40.3%, respectively) after PA than the control group. Total cell numbers in blastocyst of $10{\mu}M$ FA displayed significantly higher (39.4 vs 51.9, respectively) than the control group. In conclusion, these results suggested that treatment with FA during IVM improved the developmental potential of porcine embryos by increasing intracellular GSH synthesis and reducing ROS levels. Also, there was an improvement of cleavage rate, blastocyst formation and total cell numbers in blastocysts. It might be associated with Keap1-Nrf2 pathway as an antioxidant regulate pathway that plays a crucial role in determining the sensitivity of cells to oxidative damages by regulating the basal and inducible expression of enzymes which is related to detoxification and anti-oxidative effects, stress response enzymes and/or proteins and ABC transporters.

Association Study of Glutathione-S-Transferase M1/T1 Gene Polymorphism with Deficiency-Excess Differentiation-syndrome in Korean Bronchial Asthmatics (한국인 기관지 천식 환자에서 허설변증과 Glutathione-S-Transferase 유전자의 다형성 연구)

  • Yu, Seung-Ryeol;Jeong, Seung-Yeon;Jung, Ju-Ho;Kim, Jin-Ju;Jung, Sung-Ki
    • The Journal of Internal Korean Medicine
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    • v.28 no.3
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    • pp.453-463
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    • 2007
  • Backgrounds : Glutathione-s-transferase (GST) is a kind of phase II metabolism enzyme and plays an important role in the detoxification of various toxic chemicals. It was reported that the genetic polymorphism of GSTM1 and GSTT1 genes may be responsible for asthma development and susceptibility to allergy. Traditional oriental medicine uses a unique diagnostic technique. differentiation-syndrome. to analyze signs and symptoms of patients synthetically. Through differentiation-syndrome. asthma patients can be divided into two groups: the deficiency syndrome group (DSG) and the excess syndrome group (ESG). Objectives : The purpose of this study was to investigate the possible association of GST gene polymorphism with clinical phenotype by differentiation-syndrome of bronchial asthma patients. Materials and Methods : One hundred and ten participants were evaluated by pulmonary function test. Patients with 53 DSG and 31 ESG by differentiation-syndrome were assessed for genetic analysis. GSTM1 and GSTT1 deletion polymorphism was performed by polymerase chain reaction (PCR). Results : GSTM1 gene deletion was detected in 43.4% of individuals in the DSG and in 38.71 % in the ESG. The distribution of GSTM1 polymorphism between DSG and ESG was not significantly different [$x^2$=0.1767, p=0.6742; OR(95% CI)=1.2139(0.4915-2.9979)]. The proportion of GSTT1 null genotypes was 41.51% in the DGS and 45.16% in the ESG. The distribution of GSTT1 polymorphism between DSG and ESG was also not significantly different [$x^2$=0.1065, p=0.7442; OR(95% CI)=0.8618(0.3525-2.1065)]. In the combined analysis of GSTM1 and GSTT1 genes, the frequency of both null type of GSTM1/GSTT1 genes was not significantly different from both positive type of GSTM1/GSTT1 genes[$x^2$=0.0768, p=0.7817; OR(95% CI)=1.2000(0.3303-4.3602)] Conclusions : These results indicate that polymorphism of the GST gene might not be associated with the symptomatic classification of DSG and ESG by differentiation-syndrome in Korean asthmatics.

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Arsenic Concentrations of Groundwater and Rice Grains in Bangladesh and Phytoremediation (방글라데시의 지하수와 쌀의 비소오염 및 식물정화법)

  • Islam, Jahidul Mohammad;Kim, Bomchul;Laiju, Nahida;Nasirullah, Tarek;Miah, Mohammad Nuruddin
    • Journal of Korean Society on Water Environment
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    • v.26 no.1
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    • pp.116-124
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    • 2010
  • While groundwater is the major source for drinking and irrigation purposes, arsenic (As) contamination of groundwater is a serious issue in Bangladesh. With a view to reduce As contamination in drinking water the guideline value recommended for Bangladesh is 0.05 mg/L. We assessed groundwater As in an As-affected Sadar Upazilla (small administrative unit) in the District (administrative unit) of Chapai Nabwabganj during 2006, where 50% hand tube well water were above the recommended limit (0.05 mg/L) during dry season. Almost 20% tube well waters were above the recommended limit during rainy season, perhaps due to the dilution of water table. The groundwater in Bangladesh contaminates surface soils and plants thereby As entering the food chain. In 2005, we examined the As levels in different rice varieties grown in different Districts of Bangladesh and the As concentrations in rice grain ranged from 0.07~1.12 mg/kg while the concentrations in 3 rice varieties were above the recommended limit (1 mg/kg rice grain) and the maximum concentration was 1.12 mg/kg rice grain in the rice variety BR 11. With few exceptions, the As content of rice grain in Bangladesh is not considered to be concentration of greater health concern as yet. We also observed enhanced root uptake, efficient root-to shoot translocation, and a much elevated tolerance through internal detoxification all contribute to As hyperaccumulation in a plant, ladder brake fern (Pteris vittata L.). But the phytoremediation technique might not be an appropriate tool to reduce the As calamity in the vast areas of Bangladesh. To mitigate the As problem of Bangladesh, better coordination among governmental agencies and many other organizations will be required to combat the disaster.

Detoxification of Glutaraldehyde Treated Porcine Pericardium Using L-arginine & $NABH_4$

  • Kim, Kwan-Chang;Kim, Soo-Hwan;Kim, Yong-Jin
    • Journal of Chest Surgery
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    • v.44 no.2
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    • pp.99-107
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    • 2011
  • Background: Calcification is the most frequent cause of clinical failure of bioprosthetic tissues fabricated from GA-fixed porcine valves or bovine pericardium. A multi-factorial approach using different mechanisms was recently developed to reduce the calcification of bioprosthetic tissues. The purpose of the present study was to evaluate the synchronized synergism of using L-arginine and $NaBH_4$, compared with ethanol and L-lysine, in glutaraldehyde treated porcine pericardium from the standpoint of calcification and tissue elasticity. Materials and Methods: Porcine pericardium was fixed at 0.625% GA (7 days at room temperature after 2 days at $4^{\circ}C$). An interim step of ethanol (80%; 1 day at room temperature) or L-lysine (0.1 M; 2 days at $37^{\circ}C$) or L-arginine (0.1 M; 2 days at $37^{\circ}C$) was followed by completion of the GA fixation. A final step of NaBH4 (0.1 M; 2 days at room temperature) was followed. Their tensile strength, thickness, and thermal stability were measured. Treated pericardia were implanted subcutaneously into three-week-old Sprague-Dawley rats for 8 weeks. Calcium content was assessed by atomic absorption spectroscopy and histology. Results: L-arginine and $NaBH_4$ pretreatment ($1.81{\pm}0.39$ kgf/5 mm p=0.001, $0.30{\pm}0.08$ mm p<0.001) significantly increased tensile strength and thickness compared with the control ($0.53{\pm}0.34$ kgf/5 mm, $0.10{\pm}0.02$ mm). In a thermal stability test, L-arginine and $NaBH_4$ pretreatment ($84.25{\pm}1.12^{\circ}C$, p=0.023) caused a significant difference from the control ($86.25{\pm}0.00^{\circ}C$). L-lysine and $NaBH_4$ pretreatment ($183.8{\pm}42.6$ ug/mg, p=0.804), and L-arginine and $NaBH_4$ pretreatment ($163.3{\pm}27.5$ ug/mg, p=0.621) did not significantly inhibit calcification compared to the control ($175.5{\pm}45.3$ ug/mg), but ethanol and $NaBH_4$ pretreatment did ($38.5{\pm}37.3$ ug/mg, p=0.003). Conclusion: The combined pretreatment using L-arginine and $NaBH_4$ after GA fixation seemed to increase the tensile strength and thickness of porcine pericardium, fixed with GA. Additionally, it seemed to keep thermal stability. However it could not decrease the calcification of porcine pericardium fixed with GA. $NaBH_4$ pretreatment seemed to decrease the calcification of porcine pericardium fixed with GA, but only with ethanol.

In vivo Metabolism of Endosulfan in Carp (cyprinus carpio L.) (In vivo 시험에 의한 잉어(cyprinus carpio L.)체내 endosulfan의 대사)

  • Lee, K.B.;Shim, J.H.;Suh, Y.T.
    • Applied Biological Chemistry
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    • v.37 no.3
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    • pp.194-202
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    • 1994
  • To study the metabolism and absorption of endosulfan in carp, $^{14}C-{\alpha}-endosulfan$ was treated with the $LC_{10}$ concentration $(4.5\;{\mu}g/L)$. In an in vivo test, endosulfan was metabolized $(65{\sim}80%)$ in tissues and endosulfan ether, endosulfan alcohol, endosulfan ${\alpha}-hydroxyether$, and endosulfan lactone were identified, indicating that those are the main metabolites of detoxification in carp. The maximum levels of $^{14}C-endosulfan$ in the head, muscle, and gut occurred after 8 hr exposure. However, the maxima reached in the liver and kidneys after 30 min and 4 hr, respectively. Radioactivity in the tissue decreased rapidly 8 hr after treatment. The total amount of $^{14}C-endosulfan$ recovered in the liver, kidneys and gut of fish was about $80{\sim}90%$ during the 8 hr treatment. The non-extractable radioactivity increased after 8 hr exposure $(27{\sim}31%)$. Endosulfan sulfate, the main degradation product in plant and mouse, was not detected during the test interval from tissues of the carp.

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Response of Antioxidative Enzymes of Two Rice Cultivars to Ozone Exposure and Nutrient Supply

  • Lee, Sang-Chul;Hwan, Cho-Jeong;Park, Shin-Young;Son, Tae-Kwon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.1
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    • pp.40-46
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    • 2001
  • Ozone ($O_3$)-induced changes in chlorophyll content and specific activities of antioxidant enzymes, such as superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) were investigated in two rice cultivars (Oryza sativa L.) grown under variable nutrient treatments. For this study, two rice cultivars of Ilpumbyeo (IL) and Keumobyeo#l (KM), which were known as resistant and susceptible to $O_3$, respectively, were exposed to $O_3$at 0.15ppm for 30 days and investigated with 10 days interval. The available nutrient regimes were varied by doubling the supply of nitrogen (N), phosphorus (P) and potassium (K) Within a basic fertilizer status (N, P, K; 15, 12, 12kg/l0a$^{-1}$ ). In both cultivars and at all nutrient status, chlorophyll content in $O_3$-treated plants decreased with prolonged treatment period, although higher N, P and K supply with $O_3$ treatment alleviated the decrease in chlorophyll content. The activities of almost all enzymes investigated for this study were decreased during initial stages of $O_3$- exposure except GPX which maintained higher activity throughout the exposure period than the non-treated plant. However, the antioxidant enzymes in $O_3$-treated plants showed almost the same or higher activities on 30 days after $O_3$ - exposure. The most significant changes in activities were observed in GR of the $O_3$-treated leaves. With the prolonged treatment period, the activity of GR at 30 days was increased by 3-8 times compared to those in 10 days. Most of the investigated enzymes showed very similar tendency to $O_3$ treatment in all fertilizer status. There was no observed evidence for enhanced detoxification of $O_3$-derived activated oxygen species in plants grown under higher fertilizer status compared with that in plants grown under basic fertilizer status. The increase in the activities of SOD, APX and GR in rice leaves by relatively long-term treatment with $O_3$ at low concentration is considered to indicate that the plant became adapted to the $O_3$ stress and the protection system increased its capacity to scavenge toxic oxygen species. Our results in two rice cultivars indicated that there was little difference in the activities of antioxidant enzymes between IL and KM, which were known as resistant and susceptible cultivar to $O_3$

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Anti-proliferative Effects of the Isothiocyanate Sulforaphane on the Growth of Human Cervical Carcinoma HeLa Cells (Sulforaphane에 의한 HeLa 인체자궁경부함세포의 증식 억제 기전 연구)

  • Park Soung Young;Bae Song-Ja;Choi Yung Hyun
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.397-405
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    • 2005
  • Sulforaphane, an isothiocyanate derived from hydrolysis of glucoraphanin in broccoli and other cruciferous vegetables, was shown to induce phase II detoxification enzymes and inhibit chemically induced mammary tumors in rodents. Recently, sulforaphane is known to induce cell cycle arrest and apoptosis in human cancer cells, however its molecular mechanisms are poorly understood. In the present study, we demonstrated that sulforaphane acted to inhibit proliferation and induce morphological changes of human cervical carcinoma HeLa cells. Treatment of HeLa cells with $10{\mu}M\;or\;15{\mu}M$ sulforaphane resulted in significant G2/M cell cycle arrest as determined by flow cytometry. Moreover, $20{\mu}M$ sulforaphane significantly induced the population of sub-G1 cells (9.83 fold of control). This anti-proliferative effect of sulforaphane was accompanied by a marked inhibition of cyclin A and cyclin-dependent kinase (Cdk)4 protein and concomitant induction of Cdc2, Cdk inhibitor p16 and p21. However, sulforaphane did not affect the levels of cyelooxygenases and telomere-regulatory gene products. Although further studies are needed, the present work suggests that sulforaphane may be a potential chemoprevetive/ chemotherapeutic agent for the treatment of human cancer cells.

The XPS and SEM Evaluation of Various Technique for Cleansing and Decontamination of The Rough Surface Titanium Implants (수종의 방법으로 임프란트 표면 처치후 표면의 형태 및 성분 변화 분석에 관한 연구)

  • Kim, Sun-bong;Yim, Sung-Bin;Chung, Chin-Hyung
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.749-763
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    • 2001
  • Osseointegrated titanium implants have become an integral therapy for the replacement of teeth lost. For dental implant materials, titanium, hydroxyapatite and alumina oxide have been used, which of them, titanium implants are in wide use today. Titanium is known for its high corrosion resistance and biocompatability, because of the high stability of oxide layer mainly consists of $TiO_2$. With the development of peri-implantitis, the implant surface is changed in surface topography and element composition. None of the treatments for cleaning and detoxification of implant surface is efficient to remove surface contamination from contaminated titanium implants to such extent that the original surface elemental composition. In this sights, the purpose of this study was to evaluate rough surface titanium implants by means of scanning electron microscopy(SEM) and X-ray photoelectron spectroscopy(XPS) with respect to surface appearance and surface elemental composition. Moreover, it was also the aim to get the base for treatments of peri-implantitis. For the SEM and XPS study, rough surface titanium models were fabricated for control group. Six experimental groups were evaluated: 1) long-time room exposure, 2 ) air-powder abrasive cleaning for 1min, 3) burnishing in citric acid(pH1) for 1min, 4) burnishing in citric acid for 3min, 5) burnishing in tetracycline for 1min, 6) burnishing in tetracycline for 3min. All experimental treatments were followed by 1min of rinsing with distilled water. The results were as follows: 1. SEM observations of all experimental groups showed that any changes in surface topography were not detected when compared with control group. (750 X magnification) 2. XPS analysis showed that in all experimental groups, titanium and oxygen were increased and carbon was decreased, when compared with control group. 3. XPS analysis showed that the level of titanium, oxygen and carbon in the experimental group 3(citric acid treatment for 1min, followed by 1min of distilled water irrigation) reached to the level of control group. 4. XPS analysis showed that significant differences were not detected between the experimental group 1 and the other experimental groups except of experimental group 3. The Ti. level of experimental group 2, airpowder abrasive treatment for lmin followed by 1min of saline irrigation, was lower than the Ti. level of tetracycline treated groups, experimental group 5 and 6. From the result of this study, it may be concluded that the 1min of citric acid treatment followed by same time of rinsing with distilled water gave the best results from elemental points of view, and can be used safely to treat peri-implantitis.

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Glucose-6-phosphate dehydrogenase deficiency does not increase the susceptibility of sperm to oxidative stress induced by H2O2

  • Roshankhah, Shiva;Rostami-Far, Zahra;Shaveisi-Zadeh, Farhad;Movafagh, Abolfazl;Bakhtiari, Mitra;Shaveisi-Zadeh, Jila
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.4
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    • pp.193-198
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    • 2016
  • Objective: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzyme defect. G6PD plays a key role in the pentose phosphate pathway, which is a major source of nicotinamide adenine dinucleotide phosphate (NADPH). NADPH provides the reducing equivalents for oxidation-reduction reductions involved in protecting against the toxicity of reactive oxygen species such as $H_2O_2$. We hypothesized that G6PD deficiency may reduce the amount of NADPH in sperms, thereby inhibiting the detoxification of $H_2O_2$, which could potentially affect their motility and viability, resulting in an increased susceptibility to infertility. Methods: Semen samples were obtained from four males with G6PD deficiency and eight healthy males as a control. In both groups, motile sperms were isolated from the seminal fluid and incubated with 0, 10, 20, 40, 60, 80, and $120{\mu}M$ concentrations of $H_2O_2$. After 1 hour incubation at $37^{\circ}C$, sperms were evaluated for motility and viability. Results: Incubation of sperms with 10 and $20{\mu}M\;H_2O_2$ led to very little decrease in motility and viability, but motility decreased notably in both groups in 40, 60, and $80{\mu}M\;H_2O_2$, and viability decreased in both groups in 40, 60, 80, and $120{\mu}M\;H_2O_2$. However, no statistically significant differences were found between the G6PD-deficient group and controls. Conclusion: G6PD deficiency does not increase the susceptibility of sperm to oxidative stress induced by $H_2O_2$, and the reducing equivalents necessary for protection against $H_2O_2$ are most likely produced by other pathways. Therefore, G6PD deficiency cannot be considered as major risk factor for male infertility.

Molecular genetic analysis of phytochelatin synthase genes in Arabidopsis

  • Ha, Suk-Bong
    • Proceedings of the Botanical Society of Korea Conference
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    • 2002.04a
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    • pp.62-72
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    • 2002
  • This study has investigated the biosynthesis and function of the heavy metal binding peptides, the phytochelatins, in plants. PCs are synthesised enzymatically from glutathione by the enzyme PC synthase in the presence of heavy metal ions. Using Arabidopsis thaliana as a model organism cadmium-sensitive, phytochelatin-deficient mutants have been isolated and characterised in previous studies. The cadl mutants have wildtype levels of glutathione, are PC deficient and lack PC synthase activity. Thus, the CADl gene has been proposed to encode PC synthase. The CADl gene was isolated by a positional cloning strategy The gene was mapped and a candidate identified. Each of four cadl mutants had a single base pair change in the candidate gene and the cadmium-sensitive, cadl phenotype was complemented by the candidate gene. This demonstrated the CADl gene had been cloned. A homologous gene in the fission yeast, Schizosaccharomyces pombe was identified through database searches. A targeted-deletion mutation of this gene was constructed and the mutant, like cadl mutants of Arabidopsis, was cadmium-sensitive and PC-deficient. A comparison of the redicted amino acid sequences reveals a highly conserved N-terminal region Presumed to be the catalytic domain and a variable C-terminal region containing multiple Cys residues proposed to be involved in activation of the enzyme by metal ions. Similar genes were also identified in animal species. The Arabidopsis CADl/AtPCSl and S. pombe SpbPCS genes were expressed in E. coli and were shown to be sufficient for glutathione-dependent, heavy metal activate PC synthesis in vitro, thus demonstrating these genes encode PC synthase enzymes. Using RT-PCR, AtPCSl expression appeared to be independent of Cd exposure. However, at higher levels of Cd exposure a AtPCSl-CUS reporter gene construct appeared to be more highly expressed. Using the reporter gene construct, AtPCSl was expressed most tissues. Expression appeared to be greater in younger tissues and same higher levels of expression was observed in some regions, including carpels and the base of siliques. AtPCS2 was a functional gene encoding an active PC synthase. However, its Pattern of expression and the phenotype of a mutant (or antisense line) have not been determined. Assuming the gene is functional then it has clearly been maintained through evolution and must provide some selective advantage. This implies that, at least in some cells or tissue, it is likely to be the dominant PC synthase expressed. This remains to be determined

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