Protection against ischemia-reperfusion injury is crucial for successful transplantation of the lung. It has been known that nitric oxide has many favorable effects on the donor lungs but at the same time, has some potential side effects of cytotoxicity. In this regards, we investigated whether the administration of nitroglycerin could decrease ischemia-reperfusion injury in isolated rat lung reperfusion model for the confirmation of the effect of nitroglycerin, a donor of nitric oxide, on lung transplantation. Material and Method: 35 Sprague-Dawley species male white rats were used for this experiment. For nitroglycerin group (n=18), nitroglycerin was administered intravenously followed by mixed in flushing solution for preservation. As a control group (n=17), we used the same amount of normal saline. To evaluate the effect of nitroglycerin on the lung, heart-lung block was obtained, weighed and stored in University of Wisconsin Solution at 1$0^{\circ}C$ for 24 hours. In each group of the isolated lungs, reperfusion was carried out with Krebs-Hensleit-diluted human blood for 60 minutes. As parameters of the state of the isolated lung, peak inspiratory and pulmonary arterial pressures were continuously recorded. Oxygen and carbon dioxide tension of reperfusing blood were measured before and after 30, 60 minutes of reperfusion. After sixty minutes of reperfusion, protein content in bronchoalveolar lavage fluid was measured also for the evaluation of the degree of alveolar flooding. Lung myeloperoxidase activity was determined to verify the accumulation of neutrophils. Results: Although statistically significant differences were not noted in peak inspiratory and pulmonary arterial pressure between control and nitroglycerin group, latter group showed lowering tendency of pulmonary arterial pressure during the entire reperfusion period. Oxygen tension was higher (p<0.05) in nitroglycerin group compared with that of the control group, in contrast, there were no differences in carbon dioxide tension, protein content in bronchoalveolar lavage fluid and myeloperoxidase activity between the groups. In the examination of ultrastructural changes, nitroglycerin denoted the protective effect on the pulmonary architecture compared with that of control group. Conclusion: Collectively, on the bases of these experimental results, prior treatment of donor lung with nitroglycerin could result in better preservation of the lung. Consequently, these nitroglycerin preserved lungs are thought to be more suitable for successful transplantation of the lung.
Journal of the Korean Society of Food Science and Nutrition
/
v.40
no.10
/
pp.1353-1360
/
2011
This study was performed to investigate the antimicrobial effects against food-borne pathogens and antioxidant activity of Rhododendron brachycarpum ethanol-extract. The antimicrobial activity of the extract was determined using a paper disc-diffusion method, and the diameter of the clear zone was measured. The diameter of the clear zone in the presence of 10 mg of extract was maximal against Bacillus cereus among the three tested Gram-positive bacteria and against Escherichia coli O157:H7 among the five tested Gram-negative bacteria. Analysis of the minimum inhibitory concentration (MIC) showed that the extract exhibited a similar efficacy as that of sorbic acid, a well-known chemical preservative. The growth inhibitory effects of the extract at concentrations of 250, 500, 1,000, and 2,000 mg/L on food-borne pathogens were determined against Staphylococcus aureus, Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7. Growth of the microorganisms was not affected by the extract at concentrations up to 250 mg/L, but it was significantly (p<0.05) inhibited by the extract at concentrations higher than 1,000 mg/L. The antioxidant effects of the extract were examined via measurement of DPPH radical scavenging activity, inhibition of reactive oxygen species (ROS) generation using fluorescent dichlorofluorescien (DCF) assay, and prevention of peroxyl radical- and hydroxyl radical-induced supercoiled DNA breakage. The $IC_{50}$ of the extract for DPPH radical scavenging activity was about half that of ${\alpha}$-tocopherol, which was used as a positive control. DCF fluorescence intensity decreased as the concentration of the extract increased, demonstrating that ROS generation was inhibited in a concentration-dependent manner. The ROS inhibitory effect of the extract was higher than that of ascorbic acid. The extract prevented supercoiled DNA strand breakage induced by peroxyl radical and hydroxyl radical. Thus, the results of the present study demonstrate that the extract exhibits antimicrobial effects against food-borne pathogens as well as potent antioxidant capacity, suggesting that R. brachycarpum could be used as a natural antibacterial agent and effective antioxidant in food.
Purpose: To evaluate clinical results of a single percutaneous injection of platelet-rich plasma in patients with refractory lateral epicondylitis. Materials and Methods: Between Jan and Dec 2009, fifteen patients (5 male, 10 female) received a diagnosis of lateral epicondylitis of the elbow and were evaluated in this study. Their average age was 43.5 years. All patients were initially given a variety of non-surgical treatments for more than 1year. All patients were considering surgery. These patients were given a single percutaneous injection of 3cc of platelet-rich plasma. To assess pain, we used a visual analogue scale (VAS) at rest and during work & the Patient-Rated Tennis Elbow Evaluation (PRTEE) score. We compared the score before treatment with scores 4 and 12 weeks after treatment. Results: Average VAS scores at rest improved from 4.6 before treatment to 2.5 at week 4, and 1.8 at week 12. The average VAS score while working also improved from 7.8 before treatment to 6.2 at week 4, and 4.25 at week 12. The average PRTEE score improved from 60.13 before treatment to 46.12 at week 4 and 24.6 at week 12. Conclusion: Treatment using a single percutaneous injection of platelet-rich plasma in patients with refractory lateral epicondylitis appears to be an effective treatment modality. Platelet-rich plasma should be considered before surgical intervention.
Acute respiratory distress syndrome (ARDS), also known as an acute inflammatory lung disease is developed by various factors that is originated from the destruction of alveolar-capillary barrier, and neutrophils plays an important role in the destruction. The study intended to confirm, the anti-inflammatory effect of germanium, whether a lung injury has been mitigated with the reduction of injury in alveolar-capillary barrier resulting from inhibition of neutrophils migration in lung tissue. Test groups were divided in saline administered CON, 5 hours of endotoxin administered LPS and 5 hours of endotoxin administered Ge+LPS following 1 hours of pre-processed germanium. $100{\mu}g$ endotoxin was melted in 0.5 mL saline and sprayed into airway and 26 mg germanium per 100 g weight was administered into abdominal cavity. The endotoxin group which induced an acute lung injury with administered endotoxin showed dramatic increase of pulmonary edema (p<0.001), protein contents in bronchoalveolar lavage fluid, BALF (p<0.05) and neutrophils of infiltration in BALF (p<0.001) comparing with a control group, while a pre-treated germanium group showed significant decrease in all categories comparing to the endotoxin administerd group. In the result of a microscopic observation, the structure of alveolar-capillary barrier which is constructed with basal lamina, alveolar type I cells and endothelial cell were preserved of the pre-treated germanium group relatively well compare to the endotoxin administered group. And the construction of lamellar body, microvilli and basal lamina of alveolar type II cells were also preserved relatively well. Hence, germanium activates as an anti-Inflammatory mediator in other words, it interfered neutrophils migration into lung tissue, thereby reduced injury of alveolar-capillary barrier from toxic substances of activated neutrophils. Consequently, the study has determined that the acute lung injury induced by endotoxin has been decreased by the pre-treated germanium.
This study was to investigate the effect of chitosan-ascorbate (CA) prepared with different molecular weight (223, 746, 1110 and 2025 kDa) on the fermentation and quality of mul-kimchi, and also, the effect of the mul-kimchi juice fermented with 0.1% chitosan-ascorbate prepared with 2025 kDa chitosan (CA2025) on the serum lipids of rae fed with high cholesterol diet (HC). The mul-kimchi fermented in 0.1% CA2025 has lower turbidity, longer shelf-life, higher glucosamine content and SOD activity than those of control and the other treatments. In the animal experiment the HC-CA2025 and HC-2CA2025 groups which were administered 0.5 mL per 100 g body weight of mul-kimchi juice fermented with 0.1% CA2025 and administered the same volume of the juice concentrated 112 per nay fed for 5 week, respectively. The average body weight of the HC-CA2025 and HC-2CA2025 group was lower 6.9% and 8.4% than that of HC control group respectively. Total serum cholesterol contents of these groups were 88.33 and 85.00 mg/uL, which were 17.45% and 20.56% lower than HC control group respectively. While, HDL-cholesterol content of these groups were 7% and 23%, higher the LDL-cholesterol content were 13% and 26% lower than those of HC control group respectively. And also, though the atherogenic index, AST and ALT activities were not reached to normal control group, the values were remarkably lower than those of HC control group. This study indicates the possibility of utility for kimchi industry by adding chitosan-ascorbate, especially CA2025 showing not only quality enhancing and shelf-life prolongation but also improving serum lipids and atherogenic index in rats fed with high cholesterol diet.
Preservative effects of natural preservatives, citric acid and salt on chopped garlic were investigated. Bacterial multiplying and browning of chopped garlic were very effectively repressed by 0.5-1% citric acid. Salt had an effect on the repression of bacteria multiplying and browning of chopped garlic except for 1% NaCl. Synergistic effect between citric acid and NaCl was also very good for decreasing bacteria multiplying and maintaining Hunter color of chopped garlic. Compounded effect of the GF. citric acid. and ascorbic acid was somewhat proper in the sensory evaluation of chopped garlic. And the sensory evaluation score was the highest in chopping size 3mm(diameter) and viscosity 4500cp. of chopped garlic.
The carrier materials used for the development of bacterial inoculants to be effective in field were made with various carrier materials of two major forms, alginate bead and powder inoculants. Inoculants were prepared after mixing those carrier materials with Pseudomonas fluorescens SSL3 and Bacillus subtilis B5, and the treatment effects of each inoculants was investigated on cucumber, tomato, pepper and potato. Survival density of SSL3 and B5 in various carrier materials for duration of storage and the bead inoculants were better than the powder. In the powders, survival rate increased in carrier materials treated 5% skimilk. The growth condition of microorganisms in carrier materials is good at powder. When they were preserved in the long period, contamination is problem. Scanning(200 to 600nm) of the P. fluorescens SSL3 supernatant in centrifuged MKB broth incubated for 48h had two main peaks, pyochelin(300nm) and pyoverdin(400nm). The potato yield in field experiments of spring, treated with bead formulas showed increase of 22~29% in whole potato breeds as compared with control, because the bead formulas degraded, and released the antibiotic microorganisms in slow and constant rate. In the pot experiment, there were significant difference in soil, wheatbran, and bead formed wheatbran.
The present study was carried out to investigate the effects of maturation media on penetrability of pig oocytes by liquid boar sperm coincubated with different sperm concentrations in a modified Trisbuffered medium (mTBM). Follicular oocytes collected from ovaries of prepubertal gilts were matured in a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. The spermich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ far 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes in 5% $CO_2$in air at 38.5$^{\circ}C$, cumulus cells were removed and oocytes (30~40) were coincubated for 6 h in 0.5 $m\ell$ mTBM fertilization medium with five different (1$\times$10$^{6}$ , 2$\times$10$^{6}$ , 4$\times$10$^{6}$ , 6$\times$10$^{6}$, 10$\times$10$^{6}$$m\ell$) sperm concentrations. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ NCSU-23 culture medium fur further culture of 6 h. At 12 h after IVF, sperm penetration, polyspermy and male pronuclear formation of oocytes were evaluated. Oocytes of NCSU-23 maturation medium decreased polyspermy and increased male pronuclear formation compared to those of mTCM199 and mWaymouth MB 752/1 maturation media. Of oocytes matured in NCSU-23 medium and inseminated in mTBM medium with 2~4$\times$10$^{6}$$m\ell$ sperm concentrations, 50.8~50.9% showed sperm penetration, 13.3~19.5% polyspermy and 43.9~45.4% male pronuclear formation. In conclusion, we found out that oocytes matured in NCSU23 medium and inseminated in mTBM medium showed superior invitro fertilization compared to those matured in mTCM199 and mWaymouth MB 752/1 maturation media and inseminated in mTBM medium. The optimum sperm concentrations for in-vitro fertilization of oocytes matured in NCSU-23 medium by liquid boar semen stored at 17$^{\circ}C$ for 5 days were 2~4$\times$10$^{6}$$m\ell$.
The present study was carried out to investigate the effects of the maturation media such as a modified TCM-199 (mTCM-199) medium, modified Waymouth MB 752/1 (mWaymouth MB 752/1) medium or NCSU-23 medium on penetrability of pig oocytes by liquid boar sperm. Oocytes (30~40) were transferred into each well of a Nunc 4-well multidish containing 0.5 $m\ell$ maturation medium. When immature pig oocytes were cultured in mTCM-199, mWaymouth MB 752/1 and NCSU-23 maturation media for 44 h in 5% $CO_2$, in air at 38.5$^{\circ}C$, the germinal vesicle breakdown (CVBD) rates of the oocytes were 95.6, 94.1 and 94.9%, respectively, and the maturation rates (metaphase II) of oocytes were 92.5, 90.1 and 91.1%, respectively. No differences were observed among the maturation media. The spermrich portion of ejaculates with greater than 90% motile sperm were used in the experiment. The semen was cooled 22 to 24$^{\circ}C$ over 2 h period. The semen was diluted with Beltsville Thawing Solution (BTS) extender at room temperature to give 2$\times$10$^{8}$ sperm/$m\ell$ in 100 $m\ell$ plastic bottle. Liquid boar semen of 30 $m\ell$ in 100 $m\ell$ plastic bottle was kept at 17$^{\circ}C$ for 5 days. The sperm with greater than 70% motility after day 5 of storage were used for in-vitro fertilization (IVF). After 44 h maturation of immature oocytes, cumulus cells were removed and oocytes (30~40) coincubated far 6 h in 0.5 $m\ell$ mTCM-199 and mTBM fertilization media with 2$\times$1061$m\ell$ sperm concentration. At 6 h after IVF, oocytes were transferred into 0.5 $m\ell$ mTCM-199 and NCSU-23 culture media for further culture 6 or 42 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF, and developmental ability of oocytes at 48 h after IVF were evaluated. The oocytes in combination with NCSU-23 medium for maturation and mTBM medium for IVF increased male pronuclear formation (48.0%) compared to those in combination with mTCM-199 media for maturation and IVF, and mWaymouth MB 752il medium for maturation and mTCM-199 medium far IVF. The rates of cleaved embryos (2~4 cell stage) at 48 h after IVF were 24.1% in combination with mTCM-199 media for maturation, IVF and culture, 43.6% in combination with mWaymouth MB 75211 medium fur maturation and mTCM-199 media for IVF and culture, and 71.2% in combination with NCSU-23 medium for maturation, mTBM medium for IVF and NCSU-23 medium for culture. In conclusion, we found out the oocytes matured in vitro were fertilized by liquid boar sperm stored in BTS extender at 17$^{\circ}C$ for 5 days. We recommend the simple defined NCSU-23 medium for nuclear maturation, mTBM medium and liquid boar sperm for IVF, and NCSU-23 medium for embryo culture.
This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS + 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P<0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P<0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).
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