• Clinical and Experimental Reproductive Medicine
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• v.42 no.1
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• pp.22-29
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• 2015

Objective: Sperm must be properly prepared in in vitro fertilization (IVF)-embryo transfer (ET) programs in order to control the fertilization rate and ensure that embryos are of high quality and have appropriate developmental abilities. The objective of this study was to determine the most optimal sperm preparation method for IVF. Methods: Patients less than 40 years of age who participated in a fresh IVF-ET cycle from November 2012 to March 2013 were included in this study. Poor responders with less than three mature oocytes were excluded. Ham's F-10 medium or sperm-washing medium (SWM) was used in combination with the density-gradient centrifugation/swim-up (DGC-SUP) or SUP methods for sperm preparation. A total of 429 fresh IVF-ET cycles were grouped according to the media and methods used for sperm preparation and retrospectively analyzed (DGC-SUP/Ham's F-10, n=82; DGC-SUP/SWM, n=43; SUP/Ham's F-10, n=181; SUP/SWM, n=123). Results: There were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes. We determined that both the DGC-SUP and SUP methods for sperm preparation from whole semen, using either Ham's F-10 or SWM media, result in comparable clinical outcomes, including fertilization and pregnancy rates. Conclusion: We suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.35 no.2
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• pp.111-116
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• 2009

Gymnema sylvestre, a traditional India medicine called a 'Gagaimogouip' in Korea, has been used to regulate sugar metabolism in patients with diabetes mellitus. First, we induced callus from Gymnema sylvestre leaf and mode extract from cultured callus. In this study, we investigated the anti-obesity effect of tissue-cultured Gymnema sylvestre leaf extract in 3T3-L1 cells. The tissue-cultured Gymnema sylvestre extract inhibited the cytoplasmic lipid accumulation as well as adipogenic differentiation of preadipocytes. The tissue-cultured Gymnema sylvestre extract inhibited insulin-induced predipocyte differentiation through the inhibition of expression of the early adipogenic transcription factor C/EBP-${\alpha}$ that regulate adipogenesis. These results indicate that tissue-cultured Gymnema sylvestre extract may be potential natural ingredient for slimming cosmetic products.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.825-830
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• 2001

Escherichia coli, which harbored the ars operon from a plasmid pMH12 of Klebsiella oxytoca D12, showed filamentation due to the expression of ars genes in the presence of arsenite. The continued DNA replication in the absence of cell division was revealed, since nucleoids abound with DAPI appeared to be arranged in chains. In contrast to overexpression of arsA, its frame-shift mutant and knock-out mutant lost filamentation in the presence of arsenite, which suggested that ars-induced division block was dependent on expression of arsA. ArsA-induced division inhibition was not a consequence of an inhibition of DNA replication, and the inability of arsenite to induce an SOS response indicated that arsA-mediated division inhibition was dependent on the expression of the gene product encoded by the minB operon. ArsA is a peripheral membrane protein with an ATP-binding domain, which is homologous to MinD that requires ATP-dependent efflux. These results suggested that ArsA could possibly recruit MinC to the membrane and modulate cytoplasmic FtsZ to block assembly at the middle of the cell.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.3
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• pp.313-322
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• 1998

In a myocyte freshly isolated from rabbit cerebral artery, the characteristics of $Ca^{2+}$ release by histamine or caffeine were studied by microspectrofluorimetry using a $Ca^{2+}-binding$ fluorescent dye, fura-2. Histamine (5 ${\mu}M$) or caffeine (10 mM) induced a phasic rise of cytoplasmic free $Ca^{2+}$ concentration $([Ca^{2+}]_C)$ which could occur repetitively with extracellular $Ca^{2+}$ but only once or twice in $Ca^{2+}-free$ bathing solution. Also, the treatment with inhibitor of sarcoplasmic reticulum $Ca^{2+}-ATPase$ suppressed the rise of $[Ca^{2+}]_C$ by histamine or caffeine. In $Ca^{2+}-free$ bathing solution, short application of caffeine in advance markedly attenuated the effect of histamine, and vice versa. In normal $Ca^{2+}-containing$ solution with ryanodine (2 ${\mu}M$), the caffeine-induced rise of $[Ca^{2+}]_C$ occurred only once and in this condition, the response to histamine was also suppressed. On the other hand, in the presence of ryanodine, histamine could induce repetitive rise of $[Ca^{2+}]_C$ while the amplitude of peak rise became stepwisely decreased and eventually disappeared. These results suggest that two different $Ca^{2+}-release$ mechanisms (caffeine-sensitive and histamine-sensitive) are present in rabbit cerebral artery myocyte and the corresponding pools overlap each other functionally. Increase of $[Ca^{2+}]_C$ by histamine seems to partially activate ryanodine receptors present in caffeine-sensitive pool.

• Journal of Korean Medicine for Obesity Research
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• v.17 no.2
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• pp.77-86
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• 2017

Objectives: This study examined the effects of salvianolic acid B (SAB) on exhaustive exercise-induced cardiac muscle damage to rats. Methods: The study was carried out with 12-week-old young adult male Sprague-Dawley rats. Thirty-six rats were divided into 3 groups; normal (n=12), exhaustive exercise group (ExS, n=12) and exhaustive exercise with SAB (ExS+SAB, n=12). Five days before exhaustive exercise, SAB were medicated for 5 days in ExS+SAB group. Rats in ExS and ExS+SAB group were forced to swim for 150 minutes and then they were sacrificed, while rats in normal group were sacrificed at rest. After that, blood was collected and cardiac muscle tissue damage indices were analyzed. Results: Serum aspartate transaminase activity and lactate dehydrogenase activity were significantly lower in ExS+SAB group than in ExS group. Serum creatine phosphokinase activity of ExS+SAB was significantly lower than ExS group. However, the content of serum creatinine had no difference between ExS and ExS+SAB group. In the H&E stained left ventricle myocardium, ExS group showed signs of myocardial damage such as sporadic fragmentation of myocardial fibers, interstitial edema, cytoplasmic eosinophilia and neutrophils infiltration. However, ExS+SAB group alleviated the severity of the signs of myocardial damage. In the myocardial dihydroethidium staining, optical density was remarkably decreased in ExS+SAB group compared to ExS group. Furthermore, the up-regulation of Bax/Bcl-2 ratio was observed in ExS+SAB group compared with ExS group. Conclusions: The above results suggest that SAB may protect cardiac muscle damage via antioxidant activity and prevention of apoptosis.

• The Korean Journal of Physiology and Pharmacology
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• v.25 no.3
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• pp.239-249
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• 2021

The present study explored the therapeutic potential of hydrogen sulfide (H2S) in restoring aging-induced loss of cardioprotective effect of remote ischemic preconditioning (RIPC) along with the involvement of signaling pathways. The left hind limb was subjected to four short cycles of ischemia and reperfusion (IR) in young and aged male rats to induce RIPC. The hearts were subjected to IR injury on the Langendorff apparatus after 24 h of RIPC. The measurement of lactate dehydrogenase, creatine kinase and cardiac troponin served to assess the myocardial injury. The levels of H2S, cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), nuclear factor erythroid 2-related factor 2 (Nrf2), and hypoxia-inducible factor (HIF-1α) were also measured. There was a decrease in cardioprotection in RIPC-subjected old rats in comparison to young rats along with a reduction in the myocardial levels of H2S, CBS, CSE, HIF-1α, and nuclear: cytoplasmic Nrf2 ratio. Supplementation with sodium hydrogen sulfide (NaHS, an H2S donor) and l-cysteine (H2S precursor) restored the cardioprotective actions of RIPC in old hearts. It increased the levels of H2S, HIF-1α, and Nrf2 ratio without affecting CBS and CSE. YC-1 (HIF-1α antagonist) abolished the effects of NaHS and l-cysteine in RIPC-subjected old rats by decreasing the Nrf2 ratio and HIF-1α levels, without altering H2S. The late phase of cardioprotection of RIPC involves an increase in the activity of H2S biosynthetic enzymes, which increases the levels of H2S to upregulate HIF-1α and Nrf2. H2S has the potential to restore aging-induced loss of cardioprotective effects of RIPC by upregulating HIF-1α/Nrf2 signaling.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.5
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• pp.357-366
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• 2019

$G{\alpha}_q$-coupled receptor stimulation was implied in the activation process of transient receptor potential canonical (TRPC)1/4 and TRPC1/5 heterotetrameric channels. The inactivation occurs due to phosphatidylinositol 4,5-biphosphate ($PI(4,5)P_2$) depletion. When $PI(4,5)P_2$ depletion was induced by muscarinic stimulation or inositol polyphosphate 5-phosphatase (Inp54p), however, the inactivation by muscarinic stimulation was greater compared to that by Inp54p. The aim of this study was to investigate the complete inactivation mechanism of the heteromeric channels upon $G{\alpha}_q$-phospholipase $C{\beta}$ ($G{\alpha}_q-PLC{\beta}$) activation. We evaluated the activity of heteromeric channels with electrophysiological recording in HEK293 cells expressing TRPC channels. TRPC1/4 and TRPC1/5 heteromers undergo further inhibition in $PLC{\beta}$ activation and calcium/protein kinase C (PKC) signaling. Nevertheless, the key factors differ. For TRPC1/4, the inactivation process was facilitated by $Ca^{2+}$ release from the endoplasmic reticulum, and for TRPC1/5, activation of PKC was concerned mostly. We conclude that the subsequent increase in cytoplasmic $Ca^{2+}$ due to $Ca^{2+}$ release from the endoplasmic reticulum and activation of PKC resulted in a second phase of channel inhibition following $PI(4,5)P_2$ depletion.

• Korean Journal of Veterinary Service
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• v.42 no.2
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• pp.43-51
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• 2019

The effects on the radical scavenging activities and hepatic lipid levels in rats fed a high-fat diet (HFD) in the 70% ethanolic extract from Acer tegmentosum Maxim (ATM) were evaluated. Total phenol content of ATM was 168.60 mg catechin/g in the 70% ethanolic extract of Acer tegmentosum. The DPPH and ABTS radical scavenging activities were 18.32 mM TE/g and 32.25 mM TE/g, respectively. Food efficiency ratio was lower significantly in supplemented group with 150 mg/kg BW/day during 5 weeks (HFD+ATM) compared to HFD. Total cholesterol and triglyceride contents in liver tissue of HFD+ATM were lower significantly compared to those of the HFD. Supplementation of ATM significantly decreased lipid peroxide contents and increased radical scavenging activity in the liver tissue compared with that of HFD group. Moreover, the hepatocytes of HFD rats showed a typical fatty liver morphology showing the presence of cytoplasmic lipid droplets, whereas administration of ATM attenuated the number and the size of lipid droplets. In the liver tissue of ATM administrated HFD group, the mRNA levels of SREBP-1c, $PPAR{\gamma}$, and FAS were decreased. Therefore, these results suggest that Acer tegmentosum extracts could have antioxidant activities and the hypolipidemic effects in liver tissue by its phenolic compounds.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.1
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• pp.34-41
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• 2022

In vivo oocytes grow and mature in ovarian follicles whereas oocytes are matured in vitro in plastic culture dishes with a hard surface. In vivo oocytes show a superior developmental ability to in vitro counterparts, indicating suboptimal environments of in vitro culture. This study aimed to evaluate the influence of an agarose matrix as a culture substrate during in vitro maturation (IVM) on the development of pig oocytes derived from small antral follicles (SAFs). Cumulus-oocyte complexes (COCs) retrieved from SAFs were grown in a plastic culture dish without an agarose matrix and then cultured for maturation in a plastic dish coated without (control) or with a 1% or 2% (w/v) agarose hydrogel. Then, the effect of the soft agarose matrix on oocyte maturation and embryonic development was assessed by analyzing intra-oocyte contents of glutathione (GSH) and reactive oxygen species (ROS), expression of VEGFA, HIF1A, and PFKP genes, and blastocyst formation after parthenogenesis. IVM of pig COCs on a 1% (w/v) agarose matrix showed a significantly higher blastocyst formation, intra-oocyte GSH contents, and transcript abundance of VEGFA. Moreover, a significantly lower intra-oocyte ROS content was detected in oocytes matured on the 1% and 2% (w/v) agarose matrices than in control. Our results demonstrated that IVM of SAFs-derived pig oocytes on a soft agarose matrix enhanced developmental ability by improving the cytoplasmic maturation of oocytes through redox balancing and regulation of gene expression.

• BMB Reports
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• v.54 no.11
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• pp.563-568
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• 2021

Cancer cells predominantly generate energy via glycolysis, even in the presence of oxygen, to support abnormal cell proliferation. Suppression of PDHA1 by PDK1 prevents the conversion of cytoplasmic pyruvate into Acetyl-CoA. Several PDK inhibitors have been identified, but their clinical applications have not been successful for unclear reasons. In this study, endogenous PDHA1 in A549 cells was silenced by the CRISPR/Cas9 system, and PDHA1^WT and PDHA1^3SD were transduced. Since PDHA1^3SD cannot be phosphorylated by PDKs, it was used to evaluate the specific activity of PDK inhibitors. This study highlights that PDHA1^WT and PDHA1^3SD A549 cells can be used as a cell-based PDK inhibitor-distinction system to examine the relationship between PDH activity and cell death by established PDK inhibitors. Leelamine, huzhangoside A and otobaphenol induced PDH activity-dependent apoptosis, whereas AZD7545, VER-246608 and DCA effectively enhanced PDHA1 activity but little toxic to cancer cells. Furthermore, the activity of phosphomimetic PDHA1 revealed the complexity of its regulation, which requires further in-depth investigation.