• Journal of Veterinary Clinics
• /
• v.34 no.2
• /
• pp.156-160
• /
• 2017

We investigated the effect of ethylene glycol and antioxidants such as taurine, hypotaurine and trehalose with extenders during cryopreservation of Korean Jeju Black Bull spermatozoa. The cryopreservation of freshly collected spermatozoa was conducted with four different conditions. As a control, spermatozoa were cryopreserved with Tris egg-yolk extenders added 5% ethylene glycol (EG). Taurine (20 mM), hypotaurine (20 mM) and trehalose (20 mM) were individually added into tris egg-yolk extenders with 5% EG. After thawing of frozen spermatozoa with four different conditions, sperm viability, motility, acrosomal integrity, and membrane integrity were investigated. The significant (p < 0.05) improvement of sperm viability showed in all antioxidant treated thawed spermatozoa (taurine; $68.1%{\pm}4.4$, hypotaurine; $69.2%{\pm}6.7$ and trehalose; $68.0%{\pm}4.4$) when compared to control ($63.4%{\pm}5.6$). Neither positive nor detrimental effects of three antioxidants were shown sperm motility after thawing. The results of hypo-osmotic swelling test showed that the membrane integrity of taurine, hypotaurine or trehalose treated thawed spermatozoa ($64.1%{\pm}5.4$, $61.5%{\pm}3.7$ and $59.0%{\pm}4.0$, respectively) had significantly (p < 0.05) higher rate of the swollen sperm compared to control ($53.7%{\pm}9.7$). Hypotaurine treated frozen-thawed spermatozoa had siginificantly higher (p < 0.05) F pattern ratio than taurine, trehalose and control treated frozen-thawed spermatozoa. Trehalose added frozen-thawed spermatozoa had significantly higher (p < 0.05) acrosome reaction pattern ratio than taurine and hypotaurine added frozen-thawd spermatozoa. In this study, we found that antioxidants such taurine, hypotaurine and trehalose treatments during cryopreservation process could reduce damage of spermatozoa of Korean Jeju Black Bull and improved sperm capability of fertilization.

• Toxicological Research
• /
• v.27 no.4
• /
• pp.247-251
• /
• 2011

To clarify whether inhibitory effect of estrogen on liver tumor is associated with cell proliferation, we investigated its role in diethylnitrosamine (DEN)-induced rat preneoplastic lesions, with time sequenced manners. F344 male rats (n = 90) were divided into three groups at 5 weeks of age. The mini-osmotic pumps providing a continuous infusion of DEN was implanted into the abdominal cavity of each animal in group 1, 2 and 3 at 6 weeks of age. To see the effect of estrogen, pellet containing 1 or 10 ${\mu}g$ of estradiol-3-benzoate (EB) was implanted subcutaneously in the animals of groups 2 or 3, respectively, one week prior to DEN treatment. Ten animals of each group were euthanized at 10, 14 and 18 weeks after DEN treatment. Liver tissues at each time point were fixed in 10% phosphate-buffered formalin and were processed and embedded in paraffin and 5 ${\mu}g$ sections mounted on a silanized slide. Glutathione S-transferase placental form (GST-P) positive foci and 5-bromo-2-deoxyuridine (BrdU) labeling cells were detected at each time point. Area of GST-P positive foci in DEN+EB 1 or 10 ${\mu}g$ group was significantly decreased compared to DEN alone at 14 weeks (p < 0.01 or p < 0.05, respectively) an at 18 weeks (p < 0.05 or p < 0.01, respectively). BrdU index in DEN+EB 1 or 10 ${\mu}g$ groups was significantly decreased compared to DEN alone at 14 weeks and at 18 weeks (p < 0.01). Taken together, we conclude that EB treatment decrease the DEN-induced liver preneoplastic lesions and this may be associated with decrease of cellular proliferation.

• Journal of Chest Surgery
• /
• v.41 no.6
• /
• pp.679-686
• /
• 2008

Background: Cellular remnants in the bioprosthetic heart valve are known to be related to a host's immunologic response and they can form the nidus for calcification. The extracellular matrix of the decellularized valve tissue can also be used as a biological scaffold for cell attachment, endothelialization and tissue reconstitution. Thus, decellularization is the most important part in making a bioprosthetic valve and biological caffold. Many protocols and agents have been suggested for decellularization, yet there ave been few reports about the effect of a treatment with hypotonic solution prior to chemical or enzymatic treatment. This study investigated the effect of a treatment with hypotonic solution and the appropriate environments such as temperature, the treatment duration and the concentration of sodium dodecylsulfate (SDS) for achieving proper decellularization. Material and Method: Porcine aortic valves were decellularized with odium dodecylsulfate at various concentrations (0.25%, 0.5%), time durations (6, 12, 24 hours) and temperatures ($4^{\circ}C$, $20^{\circ}C$)(Group B). Same the number of porcine aortic valves (group A) was treated with hypotonic solution prior to SDS treatment at the same conditions. The duration of exposure to the hypotonic solution was 4, 7 and 14 hours and he temperature was $4^{\circ}C$ and $20^{\circ}C$, respectively. The degree of decellularization was analyzed by performing hematoxylin and eosin staining. Result: There were no differences in the degree of decellularization between the two concentrations (0.25% 0.5%) of SDS. Twenty four hours treatment with SDS revealed the best decellularization effect for both roups A and B at the temperature of $4^{\circ}C$, but there was no differences between the roups at $20^{\circ}C$. Treatment with hypotonic solution (group A) showed a better ecellularization effect at all the matched conditions. Fourteen hours treatment at $4^{\circ}C$ ith ypotonic solution prior to 80S treatment revealed the best decellularization effect. The treatment with hypotonic solution at $20^{\circ}C$ revealed a good decellularization effect, but his showed significant extracellular matrix destruction. Conclusion: The exposure of porcine heart valves to hypotonic solution prior to SDS treatment is highly effective for achieving decellularization. Osmotic treatment with hypotonic solution should be considered or achieving decellularization of porcine aortic valves. Further study should be carried out to see whether the treatment with hypotonic solution could reduce the exposure duration and concentration of chemical detergents, and also to evaluate how the structure of the extracellular matrix of the porcine valve is affected by the exposure to hypotonic solution.

• Korean Journal of Environmental Biology
• /
• v.18 no.2
• /
• pp.263-268
• /
• 2000

The effect of $Cd^{2+}$ on stomatal apertures of epidermal strips and intact leaves in Commelina communis L. was investigated. Cadmium stimulated stomatal opening. The stomata, treated with 100 $\mu$M $Cd^{2+}$ opened to a degree of about 8.38 fm, but the stomata, treated with no cadmium opened to 3.74 ${\mu}{\textrm}{m}$. In order to show that there was no mechanical or osmotic impediment preventing the stomata in the epidermal strips, salicylic acid (SA) and abscisic acid (ABA) were used. The treatment of 100 $\mu$M SA and 1 $\mu$M ABA inhibited 14% and 28% of stomatal opening, respectively. Other heavy metals such as $Al^{3+} and Ag^{2+}$ were also used to investigate the effect of the stomatal apertures. The treatment of $Al^{3+} and Ag^{2+}$ also stimulated 19% and 41% of stomatal opening. To understand how cadmium open stomata, the effect of cadmium on the K+ influx into the epidermal strips was investigated. $Cd^{2+}$, SA, ABA inhibited 98%, 28%, 34% of K+ uptake respectively. 3-weeks old Commelina was transferred to and grown in Hoagland solution (0,5 mM $Cd^{2+}$, 10 mM $Cd^{2+}$) for 4 days and stomatal conductance were measured. The treatment of 5 mM $Cd^{2+}$ and 10 mM $Cd^{2+}$ showed about 51% and 70% inhibition of stomatal conductance, respectively Therefore, it could be concluded that stomata in epidermal strips and intact leaves behaved differently and cadmium- stimulated stomatal opening was due to the result of cadmium uptake into the epidermal strips instead of K+. [cadmium, stomata, stomatal conductance]

• Journal of Marine Life Science
• /
• v.7 no.2
• /
• pp.205-212
• /
• 2022

Transport is essential in the farming process of farmed fish and is one of the physical stress factors such as sorting. The effect of water temperature and anesthesia during low salinity transport was confirmed. In the experimental group, the water temperature was set to 20℃ (natural water temperature, NWT), 15℃ (cooling water temperature, CWT) respectively, in water with a salinity concentration of 35‰, 15‰ and an anesthetic (anesthesia, Anes., Sigma USA) was diluted and mixed to 50 ppm. A styrofoam box (66×42×20 cm) was used as a transport container, and 8 flounder were accommodated and transported in a plastic bag injected with 3 ℓ of seawater and liquid oxygen. As a result of the study, the concentration of cortisol before transport increased significantly from 2.4±0.1 ng ml^-1 in the experimental groups except for the CWT+35‰ group (16.7±12.8 ng ml-1). The K⁺ concentration slightly increased from 3.1±0.0 mEq l^-1 before transport to 4.5±1.1 mEq l^-1 in the NWT+15‰ group, showing no difference, and significantly increased in all other experimental groups. There was no effect on changes in blood characteristics, and water temperature and anesthetic had a negative effect on osmotic control due to stress. AST and ALT were not affected.

• 한국생물공학회:학술대회논문집
• /
• 2003.10a
• /
• pp.374-377
• /
• 2003

Hybridoma cells were adapted in media containing up to 80 mM $K^+$. The adapted cells obtained tolerance at high osmotic pressure and low pH. The adapted cells showed a maximum viable cell density of $1.1{\times}10^6$ cells/ml when a batch culture was progressed in a nutrient-fortified medium with Erlenmeyer flask at 450 $mOsm/kgH_2O,$ as compared to $4.8{\times}10^5$ cells/ml for non-adapted cells grown under the same conditions. The adapted cells also showed a maximum viable cell density of $7.8{\times}10^5$ cells/ml with the same method at initial pH 7.0 as compared to $5.3{\times}10^5$ cells/ml for non-adapted cells. Adaptation of animal cells at high $K^+$ levels may therefore lead to an improvement of their performance at limited conditions.

• BMB Reports
• /
• v.30 no.6
• /
• pp.379-384
• /
• 1997

The effects of several ions on the specific activity and isozyme patterns of cellular and secretory isoperoxidases were studied in suspension-cultured cells of rice (Oryza sativa L.). Peroxidase release into the culture medium occurred in the absence of added calcium. The addition of calcium ion greatly stimulated the secretion of cationic isoperoxidases such as C2 and C3 into the medium: a maximum 11 fold increase of secretions occurred in the presence of 5 mM $CaCl_2$, and the secretion was accomplished within 1 hour after the addition of $CaCl_2$. About a 10 fold increase of the peroxidase secretion into the medium did occur with 0. 5% NaCl, whereas cellular isoperoxidase levels were reduced notably. About a 6 fold increase of the specific activity of cellular isoperoxidase was found in 5 mM $NiCl_2$-treated cell, while $NiCl_2$ had no effect on the secretion of peroxidase into the medium. Various concentrations of KCl did not change peroxidase secretion, but 5 mM $ZnCl_2$ reduced peroxidase secretion greatly. The major secretory isoperoxidases stimulated by $CaCl_2$, NaCl and cellulase were composed of cationic isoperoxidases C2 and C3, which were found to be localized in the cell wall of rice by examination of the enzyme in the protoplast. Furthermore, the secretion rates of secretory isoperoxidases were increased rapidly when cellulase was treated in the absence of the osmotic stabilizer of 0.4 M mannitol. These results suggest that the stimulations of secretory isoperoxidase levels seem to be due to the stimulation of secretion into the culture medium of rice.

• Reproductive and Developmental Biology
• /
• v.33 no.1
• /
• pp.49-54
• /
• 2009

The current study was designed to evaluate the effects of the reactive oxygen species (ROS) generated with a xanthine (X) and xanthine oxidase system (XO) on sperm function and DNA fragmentation in porcine spermatozoa. ROS were produced by a combination of $1,000{\mu}M$ X and 50 mU/ml XO. The ROS scavengers such as superoxide dismutase (SOD) (200 U/ml) and catalase (CAT) (500 U/ml) were also tested. Spermatozoa were incubated for 2 hours in BWW medium with a combination of X-XO supplemented with or without antioxidants at $37^{\circ}C$ under 5% $CO_2$ incubator. Ca-ionophore-induced acrosome reaction, the proportion of swollen spermatozoa under hypo-osmotic condition, malondialdehyde formation for the analysis of lipid peroxidation, and the proportion of DNA fragmentation were determined after 2 hours incubation. The action of ROS on porcine spermatozoa resulted in decreased Ca-ionophore-induced acrosome reaction and membrane integrity, increased the formation of malondialdehyde, and the proportion of sperm with DNA fragmentation(p<0.05). The toxic effects caused by ROS were completely alleviated by CAT in terms of sperm function and characteristics, however SOD did not serve the same scavenger effect as CAT. To conclude, the ROS can cause significant damage to porcine sperm functions and characteristics, which can be minimized by the use of antioxidants.

• Nutrition Research and Practice
• /
• v.11 no.5
• /
• pp.430-434
• /
• 2017

BACKGROUND/OBJECTIVE: Chronic hyperglycemia induces oxidative stress via accumulation of reactive oxygen species (ROS) and contributes to diabetic complications. Hyperglycemia induces mitochondrial superoxide anion production through the increased activity of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. This study aimed to determine whether fisetin and luteolin treatments suppress the oxidative stress by modulating the expression of sirtuins (SIRTs) and forkhead box O3a (FOXO3a) under hyperglycemic conditions in human monocytes. MATERIALS/METHODS: Human monocytic cells (THP-1) were cultured under osmotic control (14.5 mmol/L mannitol), normoglycemic (NG, 5.5 mmol/L glucose), or hyperglycemic (HG, 20 mmol/L glucose) conditions, in the absence or presence of fisetin and luteolin for 48 h. To determine the effect of fisetin and luteolin treatments on high glucose-induced oxidative stress, western blotting and intracellular staining were performed. RESULTS: Hyperglycemic conditions increased the ROS production, as compared to normoglycemic condition. However, fisetin and luteolin treatments inhibited ROS production under hyperglycemia. To obtain further insight into ROS production in hyperglycemic conditions, evaluation of p47phox expression revealed that fisetin and luteolin treatments inhibited p47phox expression under hyperglycemic conditions. Conversely, the expression levels of SIRT1, SIRT3, SIRT6, and FOXO3a were decreased under high glucose conditions compared to normal glucose conditions, but exposure to fisetin and luteolin induced the expression of SIRT1, SIRT3, SIRT6, and FOXO3a. The above findings suggest that fisetin and luteolin inhibited high glucose-induced ROS production in monocytes through the activation of SIRTs and FOXO3a. CONCLUSIONS: The results of our study supports current researches that state fisetin and luteolin as potential agents for the development of novel strategies for diabetes.

• Journal of the Korean Applied Science and Technology
• /
• v.32 no.3
• /
• pp.394-404
• /
• 2015

This study investigated the pharmacodynamics of betaine on the blood profile and short chain fatty acid levels in meat ducks exposed to heat wave. 400 heads of Cherry valley (Anasplatyrhynchos) meat ducks were completely randomized to 5 treatments (4 repetitions each), and were raised for 42 days. They were grouped into T1 (heat wave control group without betaine), T2 (betaine 400 ppm), T3 (betaine 800 ppm), T4 (betaine 1200 ppm), and T5 (normal control group without betaine). Compared to T1, the betaine addition groups showed higher body weight gain at shipment, with T3 showing the highest significant difference. For hematological indictors measured (red blood cells and platelets), the betaine addition groups showed significantly higher values than the heat wave control group. The pH of the former was lower but their electrolytes ($K^+$, $P^+$, and $Cl^-$) were significantly higher compared to the latter. For blood gas concentration, the former showed a significantly higher value than the latter. For the total short chain fatty acids, acetic acid, and propionic acid, the betaine addition groups and group fed broiler-high temperature diet showed higher values than the heat wave control group. On the other hand, the former showed significantly lower values in butyric acid, isobutyric acid, valeric acid, and isovaleric acid than the latter group. These results suggest that betaine has the pharmacodynamics that mediate heat stress, via the maintenance and control of the blood profile, osmotic pressure, gas concentration, and short chain fatty acid, of meat ducks under heat wave.