• Title/Summary/Keyword: Membrane damage

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Effect of Alpha-Linolenic Acid with Bovine Serum Albumin or Methyl-Beta-Cyclodextrin on Membrane Integrity and Oxidative Stress of Frozen-Thawed Boar Sperm

  • Lee, Won-Hee;Kim, Wook-Hwan;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.23 no.1
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    • pp.11-19
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    • 2019
  • The study was conducted to investigate the effects of alpha-linolenic acid (ALA) combined with bovine serum albumin (BSA) or methyl-beta-cyclodextrin (MBCD) on plasma and acrosomal membrane damages, mitochondrial activity, morphological abnormality, motility, and oxidative stress in frozen-thawed boar sperm. In previous our study, 3 ng/mL ALA had been shown protective effect during freezing process of boar sperm. Therefore, we used 3 ng/mL ALA in present study and ALA was combined with same molar ratio of BSA or MBCD (ALA+BSA and ALA+MBCD, respectively). To confirm the effect of two carrier proteins, same volume of BSA and MBCD without ALA were added during cryopreservation. Membrane damage, mitochondrial activity, reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were measured using flow cytometry, and movement of sperm tail as motility parameter and morphological abnormality were observed under light microscope. In results, all of sperm parameters were enhanced by ALA combined with BSA or MBCD compared to control groups (p<0.05). Mitochondrial activity, morphological abnormality, ROS and LPO levels in ALA+BSA or MBCD groups were no significant difference compared with ALA, BSA and MBCD treatment groups. On the other hand, plasma and acrosomal membrane intact, and sperm motility in ALA+MBCD group were higher than single treatment groups (p<0.05), whereas ALA+BSA did not differ. Our findings indicate that carrier proteins such as BSA and MBCD could improve the effect of ALA during cryopreservation of boar sperm, and treatment of ALA with carrier proteins enhance membrane integrity, mitochondrial activity through reduction of ROS-induced LPO.

Cysteine improves boar sperm quality via glutathione biosynthesis during the liquid storage

  • Zhu, Zhendong;Zeng, Yao;Zeng, Wenxian
    • Animal Bioscience
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    • v.35 no.2
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    • pp.166-176
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    • 2022
  • Objective: Sperm is particularly susceptible to reactive oxygen species (ROS) stress. Glutathione (GSH) is an endogenous antioxidant that regulates sperm redox homeostasis. However, it is not clear whether boar sperm could utilize cysteine for synthesis GSH to protect sperm quality from ROS damage. Therefore, the present study was undertaken to elucidate the mechanism of how cysteine is involved in protecting boar sperm quality during liquid storage. Methods: Sperm motility, membrane integrity, lipid peroxidation, 4-hydroxyIlonenal (4-HNE) modifications, mitochondrial membrane potential, as well as the levels of ROS, GSH, and, ATP were evaluated. Moreover, the enzymes (GCLC: glutamate cysteine ligase; GSS: glutathione synthetase) that are involved in glutathione synthesis from cysteine precursor were detected by western blotting. Results: Compared to the control, addition of 1.25 mM cysteine to the liquid storage significantly increased boar sperm progressive motility, straight-line velocity, curvilinear velocity, beat-cross frequency, membrane integrity, mitochondrial membrane potential, ATP level, acrosome integrity, activities of superoxide dismutase and catalase, and GSH level, while reducing the ROS level, lipid peroxidation and 4-HNE modifications. It was also observed that the GCLC and GSS were expressed in boar sperm. Interestingly, when we used menadione to induce sperm with ROS stress, the menadione associated damages were observed to be reduced by the cysteine supplementation. Moreover, compared to the cysteine treatment, the γ-glutamylcysteine synthetase (γ-GCS) activity, GSH level, mitochondrial membrane potential, ATP level, membrane integrity and progressive motility in boar sperm were decreased by supplementing with an inhibitor of GSH synthesis, buthionine sulfoximine. Conclusion: These data suggest that boar sperm could biosynthesize the GSH from cysteine in vitro. Therefore, during storage, addition of cysteine improves boar sperm quality via enhancing the GSH synthesis to resist ROS stress.

Fucodiphlorethol G Purified from Ecklonia cava Suppresses Ultraviolet B Radiation-Induced Oxidative Stress and Cellular Damage

  • Kim, Ki Cheon;Piao, Mei Jing;Zheng, Jian;Yao, Cheng Wen;Cha, Ji Won;Kumara, Madduma Hewage Susara Ruwan;Han, Xia;Kang, Hee Kyoung;Lee, Nam Ho;Hyun, Jin Won
    • Biomolecules & Therapeutics
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    • v.22 no.4
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    • pp.301-307
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    • 2014
  • Fucodiphlorethol G (6'-[2,4-dihydroxy-6-(2,4,6-trihydroxyphenoxy)phenoxy]biphenyl-2,2',4,4',6-pentol) is a compound purified from Ecklonia cava, a brown alga that is widely distributed offshore of Jeju Island. This study investigated the protective effects of fucodiphlorethol G against oxidative damage-mediated apoptosis induced by ultraviolet B (UVB) irradiation. Fucodiphlorethol G attenuated the generation of 2, 2-diphenyl-1-picrylhydrazyl radicals and intracellular reactive oxygen species in response to UVB irradiation. Fucodiphlorethol G suppressed the inhibition of human keratinocyte growth by UVB irradiation. Additionally, the wavelength of light absorbed by fucodiphlorethol G was close to the UVB spectrum. Fucodiphlorethol G reduced UVB radiation-induced 8-isoprostane generation and DNA fragmentation in human keratinocytes. Moreover, fucodiphlorethol G reduced UVB radiation-induced loss of mitochondrial membrane potential, generation of apoptotic cells, and active caspase-9 expression. Taken together, fucodiphlorethol G protected human keratinocytes against UVB radiation-induced cell damage and apoptosis by absorbing UVB radiation and scavenging reactive oxygen species.

The correlation of Septin4 gene expression with sperm quality, DNA damage, and oxidative stress level in infertile patients

  • Rahil Jannatifar;Hamid Piroozmanesh;Fahimeh Naghi Jalalabadi;Hamid Reza Momeni
    • Anatomy and Cell Biology
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    • v.56 no.4
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    • pp.518-525
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    • 2023
  • Septin4 belong to a family of polymerizing GTP-binding proteins that are required for many cellular functions, such as membrane compartmentalization, vesicular trafficking, mitosis, and cytoskeletal remodeling. Since, Septin4 is expressed specifically in the testis, we aimed to determine the association between Septin4 gene expression with sperm quality, DNA damage, and stress oxidative level in infertile patients. The present study included 60 semen samples that grouped into three groups: normozoospermia (n=20), asthenozoospermia (n=20), astheno-teratozoospermia (n=20). Initially, semen parameters were analyzed by using the World Health Organization protocol. The mRNA expression of Septin4 in sperm was examined using reverse transcription-polymerase chain reaction. Oxidative stress markers, i.e., total antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde, were determined by ELISA kit. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm mitochondrial membrane potential (MMP). However, it showed significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels. In conclusion, Septin4 gene expression provides clinical useful information for the diagnosis of male infertility. It might be a marker for discrimination between fertile and infertile patients. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm MMP. However, it shows significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels.

Cytotoxicity and DNA Damage Induced by Magnetic Nanoparticle Silica in L5178Y Cell

  • Kang, Jin-Seok;Yum, Young-Na;Park, Sue-Nie
    • Biomolecules & Therapeutics
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    • v.19 no.2
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    • pp.261-266
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    • 2011
  • As recent reports suggest that nanoparticles may penetrate into cell membrane and effect DNA condition, it is necessary to assay possible cytotoxic and genotoxic risk. Three different sizes of magnetic nanoparticle silica (MNP@$SiO_2$) (50, 100 and 200 nm diameter) were tested for cytotoxicity and DNA damage using L5178Y cell. MNP@$SiO_2$ had constant physicochemical characteristics confirmed by transmission electron microscope, electron spin resonance spectrometer and inductively coupled plasma-atomic emission spectrometer for 48 h. Treatment of MNP@$SiO_2$ induced dose and time dependent cytotoxicity. At 6 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$ compared to vehicle control (p<0.05 or p<0.01). Moreover, at 24 h, 50 or 100 nm MNP@$SiO_2$ decreased significantly cell viability over the concentration of 125 ${\mu}g/ml$(p<0.01). And treatment of 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Furthermore, at 48 h, 50, 100 or 200 nm MNP@$SiO_2$ decreased significantly cell viability at the concentration of 62.5 ${\mu}g/ml$ (p<0.05) and of 125, 250, 500 ${\mu}g/ml$ (p<0.01, respectively). Cellular location detected by confocal microscope represented they were existed in cytoplasm, mainly around cell membrane at 2 h after treatment of MNP@$SiO_2$. Treatment of 50 nm MNP@$SiO_2$ significantly increased DNA damage at middle and high dose (p<0.01), and treatment of 100 nm or 200 nm significantly increased DNA damage in all dose compared to control (p<0.01). Taken together, treatment of MNP@$SiO_2$ induced cytotoxicity and enhanced DNA damage in L5178Y cell.

Electrochemical Characteristics and Damage Behavior in Cathode Operating Conditions of 316L Stainless Steel with Test Time and Applied Potential in Metallic Bipolar Plates for PEMFC (고분자 전해질 연료전지 양극 작동 환경에서 실험 시간 및 작동 전압 변수에 따른 316L 스테인리스강의 전기화학적 특성과 손상 거동)

  • Shin, Dong-Ho;Kim, Seong-Jong
    • Corrosion Science and Technology
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    • v.20 no.6
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    • pp.451-465
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    • 2021
  • In this investigation, electrochemical characteristics and damage behavior of 316L stainless steel polymer electrolyte membrane fuel cell(PEMFC) were analyzed by potentiodynamic and potentiostatic tests in cathode operating condition of PEMFC. As the result of potentiodynamic polarization test, range of passive region was larger than range of active region. In the result of potentiostatic test, damage depth and width, pit volume, and surface roughness were increased 1.57, 1.27, 2.48, and 1.34 times, respectively, at 1.2 V compared to 0.6 V at 24 hours. Also, as a result of linear regression analysis of damage depth and width graph, trend lines of damage depth and width according to applied potentials were 16.6 and 14.3 times larger, respectively. This demonstrated that applied potential had a greater effect on pitting damage depth of 316L stainless steel. The damage tendency values were 0.329 at 6 hours and 0.633 at 24 hours with applied potentials, representing rapid growth in depth direction according to the test times and applied potentials. Scanning electron microscopy images revealed that surface of specimen exhibited clear pitting damage with test times and applied potentials, which was thought to be because a stable oxide film was formed by Cr and Mo.

The involvement of oxygen free radicals in the onset of aging (노화에 미치는 산소 유리라디칼에 관한 연구동향)

  • Kim, Jung-Sang;Na, Chang-Su;Kim, Young-Kon
    • Korean Journal of Oriental Medicine
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    • v.3 no.1
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    • pp.229-239
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    • 1997
  • The superoxide anion radical$(O_2)$ poses a threat to macromocules and cell organelles of the living cells. This toxicity damage to all groups of proteins results in loss of enzyme function concerned with metabolism and ion transport, and peroxidation of unsaturated fatty acids and cholesterol results in a change of permeability characteristics of the membrane, and oxidative of nucleic acids results in genomic damage and thereby cause mutation, potential carcinogenesis and somatic damage that produce cellular aging Superoxide dismutase(SOD) has received substantial attention as a potential therapeutic agent. It has been investigated as a possible agent for the prevention of ontogenesis, the reduction of cytotoxic effect of anticancer drugs, and protection against damage in ischemic tissue. It is suggest that $O_2$ is concerned with cellular aging, thereafter we need to investigate herb that activated to SOD.

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Evaluation of Creep-Fatigue Damage of KALIMER Reactor Internals Using the Elastic Analysis Method in RCC-MR

  • Koo, Gyeong-Hoi;Bong Yoo
    • Nuclear Engineering and Technology
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    • v.33 no.6
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    • pp.566-584
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    • 2001
  • In this paper, the progressive deformation and the creep-fatigue damage for the conceptually designed reactor internals of KALIMER(Korea Advanced Liquid MEtal Reactor) are carried out by using the elastic analysis method in the RCC-MR code for normal operating conditions including the thermal load, seismic load (OBE) and dead weight. The maximum operating temperature of this reactor is 53$0^{\circ}C$ and the total service lifetime is 30 years. Thus, the time- dependent creep and stress-rupture effects become quite important in the structural design. The effects of the thermal induced membrane stress on the creep-fatigue damage are investigated with the risk of the elastic follow-up. To calculate the thermal stress, detailed thermal analyses considering conduction, convection and radiation heat transfer mechanisms are carried out with the ANSYS program. Using the results of the elastic analysis, the progressive deformation and creep-fatigue damages are calculated step by step using the RCC-MR in detail. This paper ill be a very useful guide for an actual application of the high temperature structural design of the nuclear power plant accounting for the time-dependent creep and stress-rupture effects.

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Peroxidase and Photoprotective Activities of Magnesium Protoporphyrin IX

  • Kim, Eui-Jin;Oh, Eun-Kyoung;Lee, Jeong K.
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.36-43
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    • 2014
  • Magnesium-protoporphyrin IX (Mg-PPn), which is formed through chelation of protoporphyrin IX (PPn) with Mg ion by Mg chelatase, is the first intermediate for the (bacterio)chlorophyll biosynthetic pathway. Interestingly, Mg-PPn provides peroxidase activity (approximately $4{\times}10^{-2}units/{\mu}M$) detoxifying $H_2O_2$ in the presence of electron donor(s). The peroxidase activity was not detected unless PPn was chelated with Mg ion. Mg-PPn was found freely diffusible through the membrane of Escherichia coli and Vibrio vulnificus, protecting the cells from $H_2O_2$. Furthermore, unlike photosensitizers such as tetracycline and PPn, Mg-PPn did not show any phototoxicity, but rather it protected cell from ultraviolet (UV)-A-induced stress. Thus, the exogenous Mg-PPn could be used as an antioxidant and a UV block to protect cells from $H_2O_2$ stress and UV-induced damage.

Effects of Ethylacetate Fraction of Persimmon Leaves on Experimentally-induced Gastric Mucosal Damage and Gastric Ulcers in Rats

  • Choo, Myung-Hee;Park, Hyun-Suk;Shin, Kil-Man;Jung, Soon-Teck;Kim, Kyong-Su;Lee, Myung-Yul
    • Preventive Nutrition and Food Science
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    • v.5 no.1
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    • pp.37-41
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    • 2000
  • The protective effects of the ethylacetate fraction of persimmon leaves(PEF) against experimentally induced gastric mucosal damage and gastric ulcers were evaluated in ratss. In prophylatic study, 100 mg/kg ethylacetate fraction of persimmon leaves (PEFH) exhibited a total protection of 73.8% and 65.7% against HCl-ethanol and 0.2N NaOH-induced gastric mucosal membrane lesions, respectively, which was superior to cimetidine 50 mg/kg, a commonly used anti-ulcer drug. PEFH showed excellent anti-ulcer effects against pylorus ligation induced gastric ulcers, compared to the control group, however, 50 mg/kg ethylacetate fraction of persimmon leaves (PEFL) and PEFH did not affect ulcers induced by water immersion stress, and that is inferior to cimetidine 50 mg/kg. In conclusion, the results suggest that the ethylacetate fraction of persimmon leaves can be used both in prevention and treatment of experimentally induced gastric mucosal damage and ulcers.

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