• Archives of Plastic Surgery
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• v.36 no.2
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• pp.127-134
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• 2009

Purpose: Adipose tissue injection as a free graft for the correction of soft - tissue deficiency or depression deformity is a widespread procedure in plastic surgery. This study is to analyze the changes and viability of cryopreserved adipose tissue and to find out efficient long - term storage period. Methods: After centrifugation of aspirated abdominal tissues, $10m{\ell}$ of packed Adipose tissue were freezed at $-20^{\circ}C$. For 2, 4, 6, 8 months, each frozen samples were taken and injected into scalp of SCID mice. After 15 weeks, injected Adipose tissue were sampled and analyzed at 2 months interval. We compared and analyzed each group about the weight of the injected fat, histologic impressions, activity of mitochondria, size of a fat cell and rate of survival. Results: Significant weight changes were observed in cryopreservation for 2 months(p<0.05). Histologic changes were observed, independent of the freezing period with H - E stain. Among cryopreservations for 2, 4, 6 months, no significant change were observed. The reduction of mitochondrial enzymatic activity was observed independent of time interval but activity of mitochondrial dehydrogenase was reduced less than 50% in MTT assay. Conclusion: Freezing in $-20^{\circ}C$ for 6 months has no adverse effect to Adipose tissue, but fragile adipocytes, damaged cell membrane during harvesting procedure, were disrupted within 1 - 2 month and the maximum volume reduction were followed less than 2 months. These results demonstrate that tissue preparation cells without membrane damage have the greatest viability level and cryopreservation less than 2 months has great volume effect and cryopreservation for 6 months has stable volume effect.

• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.273-281
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• 2009

Antioxidants partially ameliorated the detrimental effects of reactive oxygen species (ROS) on sperm characteristics during in vitro storage. The objective of the present study was to investigate the single or synergetic antioxidative effect of curcumin and Vit. E on the characteristics of fresh boar sperm during in vitro storage. The sperm viability in curcumin, Vit. E supplementation and curcumin+Vit. $E+H_2O_2$ groups remained over 85.0% in 3 hr incubation period, but in 6 hr incubation period, curcumin+Vit. $E+H_2O_2$ groups was sharply dropped than those of curcumin and Vit. E group. The membrane intergrity in all evaluated groups except for $H_2O_2$ group did not significantly difference in 3 hr incubation period. The viability in curcumin or Vit. E supplementation were significantly increased than in curcumin+$H_2O_2$ and Vit. $E+H_2O_2$ group in 6 hr incubation period. The percentage of mitochondrial activity and acrosome intergrity obtained similar trends within same incubation periods irrespective of treatment. The lipid peroxidation of spermatozoal plasma membrane ranged from $11.6{\sim}17.5\;nM/l{\times}10^6$ and $14.0{\sim}19.0\;nM/l{\times}10^6$ in 3 hr and 6 hr incubation periods. In conclusion, curcumin or Vit. E surpplementation alone or cooperatively improved sperm viability index (motility, membrane intergrity, viability and survival rates) and fertility index (mitochondria activity, acrosome intergrity and lipid peroxidation) of fresh boar sperm, indicating that curcumin and Vit. E have a antioxidative properties through its scavenging activity against hydrogen peroxide.

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.4
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• pp.320-328
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• 1991

The present study was designed to evaluate the effects of dietary vitamin E and coenzyme $Q_{10}$ supplementation on adriamycin (ADR) -induced lipid petoxidation in rats. After feeding the experimental diets for e weeks. Ann treatment significantly decreased growth performance of rats. But this decrement was not modified by supplementation of vitamin E or coenzyme $Q_{10}$ . Lipid peroxide values of plasma and heart mitochondria were elevated by Ann treatment. But these values were significantly decreased according to vitamin E or coenzyme $Q_{10}$ supplementation. Adriamycin treatment elevated glutathione peroxidase (GSH-Px) activity of rats, but this increment was modified by vitamin E supplementation. There was a tendency of higher superoxide dismutase (SOD) activity in ADR-treated rats. However, vitamin E or coenzyme $Q_{10}$ administration reduced this enzyme activity. With ADR treatment, arachidonic acid (20 : 4) was greatly increased, but docosahexaenoic acid (22 : 6) was not detected. Arachidonic acid was decreased and docosahexaenoic acid increased by supplementation of higher level of vitamin E or coenzyme $Q_{10}$ . Present data showed that dietary vitamin E and coenzyme $Q_{10}$ influenced on ADR-induced lipid peroxidation in rats, and also the degree of antioxidative effect was greater in vitamin E-supplemented rats.

• Journal of Pharmacopuncture
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• v.18 no.2
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• pp.26-32
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• 2015

Objectives: Rheum undulatum L. has traditionally been used for the treatment of many diseases in Asia. However, its anti-proliferative activity in cancer has still not been studied. In the present study, we investigated the anti-cancer effects of methanol extract of Rheum undulatum L. (MERL) on human adenocarcinoma gastric cell lines (AGS). Methods: To investigate the anti-cancer effect of MERL on AGS cells, we treated the AGS cells with varying concentrations of MERL and performed 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assays. Cell cycle analyses, measurements of the mitochondrial membrane potential (MMP), caspase activity assays and Western blots were conducted to determine whether AGS cell death occurred by apoptosis. Results: Treatment with MERL significantly inhibited growth of AGS cells in a concentration dependent manner. MERL treatment in AGS cells leaded to increased accumulation of apoptotic sub G1 phase cells in a concentration dependent manner. In control cultures, 5.38% of the cells were in the sub G1 phase. In MERL treated cells, however, this percentage was significantly increased (9.95% at $70{\mu}g/mL$, 15.94% at $140{\mu}g/mL$, 26.56% at $210{\mu}g/mL$ and 38.08% at $280{\mu}g/mL$). MERL treatment induced the decreased expression of pro-caspase-8 and -9 in a concentration dependent manner, whereas the expression of the active form of caspase-3 was increased. A subsequent Western blot analysis revealed increased cleaved levels of poly (ADP-ribose) polymerase (PARP) protein. Also, treatment with MERL increased the activities of caspase-3 and -9 compared with the control. MERL treatment increased the levels of the pro-apoptotic truncated Bid (tBid) and Bcl2 Antagonist X (Bax) proteins and decreased the levels of the anti-apoptotic B-cell lymphoma 2 (Bcl-2) protein, whose is the stabilization of mitochondria. However, inhibitions of p38, extracellular signal regulated kinases (ERKs) and C-Jun N-terminal kinases (JNK) by MERL treatment did not affect cell death. Conclusion: These results suggest that MERL mediated cell death is associated with an intrinsic apoptotic pathway in AGS cells.

• Molecules and Cells
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• v.26 no.2
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• pp.193-199
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• 2008

The pro-apoptotic Bcl-2 family member Bim acts as a sensor for apoptotic stimuli and initiates apoptosis through the mitochondrial pathway. To identify novel regulators of Bim, we employed the yeast two-hybrid system and isolated the human gene encoding macrophage migration inhibitory factor (MIF), a ubiquitously expressed proinflammatory mediator that has also been implicated in cell proliferation, the cell cycle and carcinogenesis. The interaction between MIF and Bim was confirmed by both in vitro and in vivo protein interaction assays. Intriguingly, protein complexes between MIF and the three major Bim isoforms (BimEL/BimL/BimS) could be detected in HEK293 and K562 cells, especially in cells undergoing apoptosis. Moreover, exogenous expression of MIF partially inhibited Bim-induced apoptosis in HEK293 cells. SiRNA-mediated knockdown of MIF increased apoptosis in K562 cells exposed to the chemical oxidant diamide. Endogenous MIF may regulate the pro-apoptotic activity of Bim and inhibit the release of cytochrome c from mitochondria.

• Applied Microscopy
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• v.19 no.1
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• pp.27-33
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• 1989

Six cases of ovarian fibrous stromal neoplasm were studied clinically, light microscopically and ultrastructurally for the clinico-pathological evidences of hormone production. Of the six cases, two cases were fibroma, three cases were fibrothecoma, and one case was thecoma. Two cases of fibroma and one fibrothecoma were associated with clinical history of menstrual abnormality, however fat staining of the tumor was negative or weakly positive. Two cases of fibrothecoma and one thecoma were negative for the clinical history of hormone imbalance. Fat stain of those cases revealed positive in varying intensity. Ultrastructural examination of fibroma-thecoma group revealed dark and pale cells by their nuclear characteristics. The dark cells had indented nucleus and abundant cytoplasmic organelles of rough ER, Golgi apparatus and mitochondria. Intracytoplasmic cisternal spaces were seen in the dark cell cytoplasm and some lipid droplets were seen around the cisternae. Pale cells had pale swollen nucleus and fine chromatins. Their cytoplasm showed scanty amount of organelles. Fibroma-thecoma spectrum showed varying degree of population of dark cells, light cells and intervening collagenous stroma. Lipid droplet was structurally associated with intracytoplasmic cisterna and they were frequently seen in thecoma and two of the fibrothecoma. But clinical history of hormone imbalance was poorly related to the light microscopic morphology and ultrastructural organization.

• Applied Microscopy
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• v.33 no.3
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• pp.205-214
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• 2003

To investigate the effect of the squalene against the lead toxicity. A healthy male of ICR mice were used for experiment. The SOD was observed after the intraperitoneal injection in mice. The ultrastructural changes of the liver were observed after the intraperitoneal injection in mice. The experimental groups were divided into two groups. Group A was control group that squalene was not treated after intraperitoneal injection of lead acetate. Group B was squalene treatment group that squalene solution was injected after intraperitoneal injection of lead acetate. All groups were used to 10 mice. The results were as follow: SOD activity in the liver, Group A was lower than in normal. But, Group B was higher than in Group A (P<0.05). In the histological observation, Group A were showed that the inner cavity of mitochondria swellen and development of cristae weakened. Swelling of lamellae of rough endoplasmic reticulum (rER) was showed. It was concluded that the SQ will be effective for the recovery of hepatic cell at lead intoxication.

• Journal of Embryo Transfer
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• v.25 no.1
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• pp.35-42
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• 2010

Many studies propose that dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I) is associated with neurodegenerative disorders, such as Parkinson's disease and Huntington's disease. Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. With a recombinant adeno-associated virus vector carrying the NDI1 gene (rAAV-NDI1) as the gene delivery method, we were able to attain high transduction efficiencies even in the human epithelial cervical cancer cells that are difficult to transfect by lipofection or calcium phosphate precipitation methods. Using a rAAV-NDI1, we demonstrated that the Ndi1 enzyme is successfully expressed in HeLa cells. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced HeLa cells were not affected by rotenone which is inhibitor of complex I, but was inhibited by flavone and antimycin A. The NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. In particular, in the NDI1-transduced cells, the yeast enzyme becomes integrated into the human respiratory chain. It is concluded that the NDI1 gene provides a potentially useful tool for gene therapy of mitochondrial diseases caused by complex I deficiency.

• Journal of Food Hygiene and Safety
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• v.30 no.4
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• pp.383-389
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• 2015

This study was to examine the antioxidative activity of fermented Rhynchosia nulubilis (FRN) in obese rats. Oxidative stress due to reactive oxygen species (ROS) can cause oxidative damage to cells. Mitochondria are especially important in the oxidative stress as ROS have been found to be constantly generated as an endogen threat. Mitochondrial defense depends mainly on superoxide dismutase whereas microsomal defense depends on catalase, which is an enzyme abundant in microsomes. Seven weeks-aged female Sprague-Dawley rats were divided into four groups and fed high fat diets for 44 days. Also fermented Rhynchosia nulubilis was administered orally for 44 days at 7.5 ml/kg of body weight of rats. The antioxidative activities of fermented Rhynchosia nulubilis were measured by the superoxide dismutase, catalase, malondialdehyde levels in liver homogenate. The levels of malondialdehyde in FRN-treated groups were lower than those in obese groups. Superoxide dismutase and catalase levels were significantly increased. These results demonstrated that fermented Rhynchosia nulubilis had the inhibitive effects of oxidative stress in obese rats, suggesting that fermented Rhynchosia nulubilis would be used as an ingredient of the useful functional products.

• The Journal of Internal Korean Medicine
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• v.35 no.3
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• pp.317-331
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• 2014

Objectives: The root of Platycodon grandiflorum (PG) has been known to possess a range of pharmacological activities including anti-cancer, anti-inflammatory, and anti-oxidant effects. The present study was designed to investigate whether or not PG-induced cell death was connected with autophagy and apoptosis in NCI-H460 human lung cancer cells. Methods: Effects on the cell viability and apoptotic activity were quantified using MTT assays and flow cytometry analysis, respectively. Protein activation was measured by immunoblotting. Autophagy was measured by LC3 immunofluorescence and immunoblotting. ROS production and loss of mitochondria membrane potential (MMP) were checked with flow cytometry analysis. Results: Following exposure to PG, NCI-H460 cell proliferation decreased simultaneously inducing autophagic vacuoles and up-regulation of microtubule-associated protein 1 light chain 3 and beclin-1 protein expressions. Interestingly, pre-treated with autophagy inhibitors, 3-methyladenin or bafilomycin A1 further triggered reduction of cell viability. PG treatment also induced apoptosis that was related modulation of Bcl-2 family proteins, death receptors and activation of caspases. In addition, PG stimulation clearly enhanced loss of MMP and reactive oxygen species (ROS) generation. Conclusions: Our results suggest that PG elicited both autophagy and apoptosis by increasing loss of MMP and ROS production. PG induced-autophagy may play a cell protective role.