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Follicular fluid-derived extracellular vesicles improve in vitro maturation and embryonic development of porcine oocytes

  • Heejae Kang;Seonggyu Bang;Heyyoung Kim;Ayeong Han;Shuntaro Miura;Hye Sun Park;Islam M. Saadeldin;Sanghoon Lee;Jongki Cho
    • Korean Journal of Veterinary Research
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    • v.63 no.4
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    • pp.40.1-40.7
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    • 2023
  • To optimize the most efficient method for porcine in vitro maturation (IVM), we compared the effects of supplementing extracellular vesicles (EVs) derived from porcine follicular fluid (pFF). The cumulus oocyte complexes were grouped into 4 groups with different supplementations as following: pFF (G1), pFF-depleted EVs (G2), EVs (G3) and control (G4) groups. After IVM with different supplementations, maturation rates and the developmental competences of porcine oocytes and blastocyst development were investigated. Additionally, glutathione (GSH) and reactive oxygen species (ROS) levels were measured in mature oocytes. The EVs were isolated and characterized with cryo-TEM and nanoparticle tracking analysis. The pFF significantly affected the maturation rate, whereas the presence of EVs did not show notable difference in the maturation rates. Although there were numerical increases in the measured parameters in EV and pFF-depleted EVs groups, no significant differences were observed between them. The EV group showed similar oocyte maturation rate for both positive and negative control groups. The GSH was not different among the groups, but ROS levels were significantly lower in pFF-supplemented group when compared with other groups with the highest level in the control group. G2 group wasn't significantly different G1 and G3 group. G3 group wasn't significantly different from G2 and G4 group. This suggests that EVs in IVM medium which probably effected partially to protect against oxidative stress and potentially enhance the quality of oocytes. This study indicates that the EVs in pFF play a significant role in improving the efficiency of oocyte maturation in porcine.

Sr-Nd-Pb Isotopic Compositions of Lavas from Cheju Island, Korea (제주도 화산암류의 Sr-Nd-Pb 동위원소 연구)

  • 박준범;박계헌;정창식
    • The Journal of the Petrological Society of Korea
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    • v.5 no.1
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    • pp.89-107
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    • 1996
  • Sr, Nd and Pb isotopic characteristics of alkaline lavas and tholeiites in Cheju Island show that the isotopic compositions of the former slightly overlap, but have relatively more depleted than the latter. However, in viewpoint of the two eruptional stratigraphies of tholeiites, the isotopic compositon of the older one is similar to those of alkaline rocks in Lava Plateau Stage after Lee (1982). These suggest that the parental magmas of alkaline lavas and tholeiites might have originated from the homogenous mantle sourve and that the characteristics of the mantle source to be partially melted might be different between the eruption stages. The isotopic signatures of the bolcanic rocks in Cheju Island overlap with those in Samoa Islands and South China Basin, indicating the DMM-EM IImixing trend. This is distingushed from the DMM-EM I trend of the Cenozoic volcanic rocks in Korea except for cheju Island and Northeastern China. The modelled binary mixing calculation between MM and EM IImaterials indicates that the mantle source of the volcanic rocks in Cheju Island has been mixed about less than 10% of enriched mantle material (EM II) with depleted mantle material (DMM). Concerned with the indentation model between North China Block (NCB) and South China Block (SCB) after Yin an Nie (1993), we suggest that the distinct isotopic features of DMM-EM I and DMM-EM IIof the Cenozoic volcanic rock in Korea as well as China can be explained by the difference of the nature of subcontinental lithospheric mantle as enriched mantle materials, i.e. EM I of NCB, while EM II of SCB.

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The Effect of Dietary Calcium Level on Bone Mineral Density and Bone Mineral Content in Ovariectomized Female Rats (난소절제한 흰쥐에서 식이칼슘량이 골밀도에 미치는 영향)

  • 김경희
    • Journal of Nutrition and Health
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    • v.29 no.6
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    • pp.590-593
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    • 1996
  • This study was done to evaluate the effect of dietary calcium level (a diet which met 100% or twice the calcium level in AIN-76 diet) on preventing bone loss in ovariectomized rats. Forty Sprauge-Dawley female rats(body weight 200$\pm$5g)were divided into two groups. One group were ovariecotomized (Ovx) while the others received sham operation(Sham). Thereafter, each rat group was further divided into normal calcium diet(0.52%) and high calcium diet(1.04%) subgroups. All rats were fed on experimental diet and deionized water ad libitum for 8 weeks. The total body, spine and femur bone mineral densities and bone mineral contents were measured by Dual Energy X-ray Absorptiometry, Eight weeks following operation, ovariectomized rats fed a high calcium diet had a significantly higher total bone mineral content, total bone calcium content, spine bone mineral density, spine bone mineral content and femur bone mineral content than ovariectomized rats fed control calcium diet. The correlation between dietary calcium intake level and spine bone mineral density were positive, but there was no correlation between dietary calcium intake and femur bone mineral density. The findings from the present study demonstrated that bone loss due to ovarian hormonal deficiency can be partially prevented by a high calcium diet. Futhermore, these findings support the strategy of the use of a high calcium diet in the prevention of estrogen depleted bone loss(postmenopausal osteoporosis)

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Roles of Glutathione Reductase and $\gamma$-Glutamylcysteine Synthetase in Candida albicans

  • Baek, Yong-Un;Yim, Hyung-Soon;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.61-61
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    • 2003
  • We have cloned the CGR1 gene encoding glutathione reductase (GR) which catalyzes the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH) from Candida albicans. The cgr1/cgr1 mutants were not viable when CaMAL2 promoter repressed the CGR1 expression. The growth of the mutants could be partially overcome by thiol compounds such as GSH, dithiothreitol, cysteine, N-acetylcysteine and GSSG. Interestingly, C. albicans with CGR1 overexpressed showed defective hyphal growth on solid medium and attenuated virulence. We have also cloned the GCS1 gene encoding ${\gamma}$-glutamylcysteine synthetase which catalyzes the first step of glutathione biosynthesis. The gcs1/gcs1 mutants were nonviable in minimal defined medium. The growth of the mutants could be resumed by supplementing with GSH, GSSG and ${\gamma}$-glutamylcysteine in the medium. The mutants had increased intracellular D-erythroascorbic acid level up to 2.25-fold when transferred to GSH-free medium. When the mutants were depleted of GSH, they showed typical markers of apoptosis. In conclusion, these results suggest that glutathione is an essential metabolite, and involved in hyphal growth, virulence and apoptosis in C. albicans.

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Methylglyoxal Induces Mitochondrial Dysfunction and Cell Death in Liver

  • Seo, Kyuhwa;Ki, Sung Hwan;Shin, Sang Mi
    • Toxicological Research
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    • v.30 no.3
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    • pp.193-198
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    • 2014
  • Degradation of glucose is aberrantly increased in hyperglycemia, which causes various harmful effects on the liver. Methylglyoxal is produced during glucose degradation and the levels of methylglyoxal are increased in diabetes patients. In this study we investigated whether methylglyoxal induces mitochondrial impairment and apoptosis in HepG2 cells and induces liver toxicity in vivo. Methylglyoxal caused apoptotic cell death in HepG2 cells. Moreover, methylglyoxal significantly promoted the production of reactive oxygen species (ROS) and depleted glutathione (GSH) content. Pretreatment with antioxidants caused a marked decrease in methylglyoxal-induced apoptosis, indicating that oxidant species are involved in the apoptotic process. Methylglyoxal treatment induced mitochondrial permeability transition, which represents mitochondrial impairment. However, pretreatment with cyclosporin A, an inhibitor of the formation of the permeability transition pore, partially inhibited methylglyoxal-induced cell death. Furthermore, acute treatment of mice with methylglyoxal increased the plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), indicating liver toxicity. Collectively, our results showed that methylglyoxal increases cell death and induces liver toxicity, which results from ROS-mediated mitochondrial dysfunction and oxidative stress.

Contractile Mechanisms of Serotonin in the Renal Arterial smooth muscle of a Rabbit (Serotonin에 의한 가토 신동맥 평활근 수축기전)

  • Lee, Woo-Young;Kim, Se-Hoon;Chang, Seok-Jong
    • The Korean Journal of Physiology
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    • v.24 no.1
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    • pp.67-76
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    • 1990
  • The contractile mechanisms of serotonin were investigated in the renal artery of a rabbit. The helical strips of isolated renal artery were immersed in the normal or $Ca^{2+}$-free tris-buffered Tyrode's solution, which was equilibrated with 100% $O_{2}$ at $35^{\circ}C$. The contraction by serotonin or norepinephrine (NE) began at $1{\times}10^{-7}\;M$ and reached the maximal contraction at $1{\times}10^{-5}\;M$. The maximal contraction by serotonin corresponded to $58.1{\pm}4.2%$ of maximal contraction by NE. Cyproheptadine, a serotonin receptor blocker, shifted the concentration-response curve to the right without any reduction in the maximum response but shifted that of NE to the right with reduction in maximum response. And phentolamine, an ${\alpha}-receptor$ blocker, shifted the concentration-response curve of serotonin or NE without any reduction in maximum responses. The $pA_{2}$ values for cyproheptadine against serotonin and NE were $10.35{\pm}0.04$ and $8.45{\pm}0.13$, respectively. The $pA_{2}$ values for phentolamine against serotonin and NE were $6.87{\pm}0.04$ and $8.14{\pm}0.08$, respectively. after the pretreatment with 6-hydroxydopamine, the contraction induced by 100 mM $K^{+}$, tyramine and serotonin reduced to $83.0{\pm}2.0$, $26.8{\pm}6.2$ and $82.0{\pm}3.5%$ of control, respectively. The contraction by serotonin in the $Ca^{2+}$-free Tyrode's solution was increased and sustained with the addition of $Ca^{2+}$ extracellulary. The serotonin-sensitive intracellular $Ca^{2+}$ pool was depleted completely by the pretreatment with NE, but the NE-sensitive intracellular $Ca^{2+}$ pool was depleted partially by the pretreatment with serotonin. From the above results, it is suggested that the contraction induced by serotonin in the renal artery of a rabbit may be due to mechanisms in which serotonin acts directly on specific serotonin receptors and also acts indirectly on ${\alpha}-adrenoceptors$ by displacing NE from neuronal stores.

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Relatoinship between Sarcoplasmic Reticular Calcium Release and $Na^+-Ca^{2+}$ Exchange in the Rat Myocardial Contraction

  • Kim, Eun-Gi;Kim, Soon-Jin;Ko, Chang-Mann
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.3
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    • pp.197-210
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    • 2000
  • Suppressive role of $Na^+-Ca^{2+}$ exchange in myocardial tension generation was examined in the negative frequency-force relationship (FFR) of electric field stimulated left atria (LA) from postnatal developing rat heart and in the whole-cell clamped adult rat ventricular myocytes with high concentration of intracellular $Ca^{2+}$ buffer (14 mM EGTA). LA twitch amplitudes, which were suppressed by cyclopiazonic acid in a postnatal age-dependent manner, elicited frequency-dependent and postnatal age-dependent enhancements after $Na^+-reduced,\;Ca^{2+}-depleted$ (26 Na-0 Ca) buffer application. These enhancements were blocked by caffeine pretreatment with postnatal age-dependent intensities. In the isolated rat ventricular myocytes, stimulation with the voltage protocol roughly mimicked action potential generated a large inward current which was partially blocked by nifedipine or $Na^+$ current inhibition. 0 Ca application suppressed the inward current by $39{\pm}4%$ while the current was further suppressed after 0 Na-0 Ca application by $53{\pm}3%.$ Caffeine increased this inward current by $44{\pm}3%$ in spite of 14 mM EGTA. Finally, the $Na^+$ current-dependent fraction of the inward current was increased in a stimulation frequency-dependent manner. From these results, it is concluded that the $Ca^{2+}$ exit-mode (forward-mode) $Na^+-Ca^{2+}$ exchange suppresses the LA tension by extruding $Ca^{2+}$ out of the cell right after its release from sarcoplasmic reticulum (SR) in a frequency-dependent manner during contraction, resulting in the negative frequency-force relationship in the rat LA.

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Measurement of Gas-Accessible PCE Saturation in Unsaturated Soil using Gas Tracers during the Removal of PCE (토양 내 PCE 제거과정에서 가스 분배추적자기법을 이용한 공기노출 PCE의 잔류량 검출)

  • Kim, Heon-Ki;Kwon, Han-Joon;Song, Young-Soo
    • Journal of Soil and Groundwater Environment
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    • v.16 no.5
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    • pp.42-52
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    • 2011
  • In this laboratory study, the changes in gas-exposed perchloroethene (PCE) saturation in sand during a PCE removal process were measured using gaseous tracers. The flux of fresh air through a glass column packed with PCEcontaminated, partially water-saturated sand drove the removal of PCE from the column. During the removal of PCE, methane, n-pentane, difluoromethane and chloroform were used as the non-reactive, PCE-partitioning, water-partitioning, and PCE and water-partitioning tracers, respectively. N-pentane was used to detect the PCE fraction exposed to the mobile gas. At water saturation of 0.11, only 65% of the PCE was found to be exposed to the mobile gas prior to the removal of PCE, as calculated from the n-pentane retardation factor. More PCE than that detected by n-pentane was depleted from the column due to volatilization through the aqueous phase. However, the ratio of gas-exposed to total PCE decreased on the removal of PCE, implying gas-exposed PCE was preferentially removed by vaporization. These results suggest that the water-insoluble, PCE-partitioning tracer (n-pentane in this study), along with other tracers, can be used to investigate the changes in fluid (including nonaqueous phase liquid) saturation and the removal mechanism during the remediation process.

Reliability Analysis of SiGe pMOSFETs Formed on PD-SOI (PD-SOI기판에 제작된 SiGe p-MOSFET의 신뢰성 분석)

  • Choi, Sang-Sik;Choi, A-Ram;Kim, Jae-Yeon;Yang, Jeon-Wook;Han, Tae-Hyun;Cho, Deok-Ho;Hwang, Young-Woo;Shim, Kyu-Hwan
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2007.06a
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    • pp.533-533
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    • 2007
  • The stress effect of SiGe p-type metal oxide semiconductors field effect transistors(MOSFETs) has been investigated to compare device properties using Si bulk and partially depleted silicon on insulator(PD SOI). The electrical properties in SiGe PD SOI presented enhancements in subthreshold slope and drain induced barrier lowering in comparison to SiGe bulk. The reliability of gate oxides on bulk Si and PD SOI has been evaluated using constant voltage stressing to investigate their breakdown (~ 8.5 V) characteristics. Gate leakage was monitored as a function of voltage stressing time to understand the breakdown phenomena for both structures. Stress induced leakage currents are obtained from I-V measurements at specified stress intervals. The 1/f noise was observed to follow the typical $1/f^{\gamma}$ (${\gamma}\;=\;1$) in SiGe bulk devices, but the abnormal behavior ${\gamma}\;=\;2$ in SiGe PD SOI. The difference of noise frequency exponent is mainly attributed to traps at silicon oxide interfaces. We will discuss stress induced instability in conjunction with the 1/f noise characteristics in detail.

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Inhibition Mechanism of Endothelin-l-induced $Ca^{2+}$ Mobilization of Antimelanogenic Ingredient: 1,2-Ο-Diferulylglycerol

  • Lee, K. M.;Park, J. B.
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.73-86
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    • 2003
  • Endothelins secreted from keratinocytes are intrinsic madiators for human melanocytes in UVB-induced pigmentation. Antimelanogenic ingredient, 1,2-Ο-diferulylglycerol(SM709) isolated from bamboo extract inhibited the melanin synthesis of Bl6F10 melanoma cells by 62%. To understand the cellular mechanism of antimelanogenic activity of SM709 in human melanocytes, the effects of SM709 on the ET-l-induced $Ca^{2+}$ mobilization were investigated. ET-l receptors in human melanocytes were characterized by using specific antagonist and found that ET-l increased intracellular $Ca^{2+}$ by activating ET-B receptor. SM709 completely blocked the ET-l-induced intracellular $Ca^{2+}$ increase and its inhibitory effect showed dose- and time- dependent manners. To investigate the role of SM709 on intracellular $Ca^{2+}$ store, when the $Ca^{2+}$ store was partially depleted by thapsigargin; a specific inhibitor of ER-type $Ca^{2+}$-ATPase, caffeine-induced $Ca^{2+}$ mobilization did not changed in the presence or absence of SM709, suggesting that SM709 has no effect on the $Ca^{2+}$ store. It is known that LPA receptor and P$_2$ receptor are linked to InsP$_3$ second messenger system. When these receptors in melanocytes were activated by LPA and ATP, the intracellular $Ca^{2+}$ signaling was observed even in the presence of SM709. From the above results, it can be suggested that SM709 has an antimelanogenic activity by antagonizing the ET-B receptor, resulting in subsequent intracellular $Ca^{2+}$ signaling, in UV induced pigmentation.nduced pigmentation.

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