• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.2
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• pp.79-92
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• 1992

This study was carried out to develop new techniques useful for cryopreservation, thawing and artificial insemination, and ultrastructural changes of cryopreserved spermatozoa in rainbow trout(Oncorhynchus mykiss) . Two extenders, such as Tyrode solution and Whittingham's $T_6$ solution, were used to preserve rainbow trout sperm in refrigerator $(-20,\;-40\;and\;-70^{\circ}C)$ or liquid nitrogen $%(-196^{\circ})$. Hand-stripped semen was diluted to 1:16 with two extenders, an then the semen were frozen after mixing semen and each extender containing 1M or 1.5M DMSO solution to 1:1. After 60 days cryopreserved semen was thawed in a $13^{\circ}$ water bath, and subsequently centrifugated. After centrifugation at 1,000 rpm for 5 min thawed semen was washed with extenders, and then fertilized with fresh eggs. The results obtained in these experiments were summarized as follows: After cryopreservation, over 75% of spermatozoa were appeared motile and the survival rate was high. Following cryopreservation by the addition of cryoprotectant such as DMSO, methanol and glycerol, the fertilization rate of the thawed spermatozoa appeared over $99\%$ compared with the control having $99\%$ of fertilization rate. There was no difference between the control and experimental groups such as $(-20^{\circ}C\;-40^{\circ}C\;and\;-70^{\circ}C)$ and $-196^{\circ}$ in fertilization rate. Following cryopreservation at $-196^{\circ}$ by the addition of 1M DMSO of cryoprotectant, each fertilization rate following 24 hours and hatching rate following 24 days showed $96\%$ and $8\%$ by the addition of BSA, but showed $98\%\;and\;10%$ by no addition of BSA. Following 2 months of cryopreservation by the addition of 1M DMSO of cryoprotectant, there were $10%$ of hatching rate at $-196^{\circ}\;and\;10\%\;and\;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1M methanol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C,\;and\;28\%,\;at\;-70^{\circ}C$ Following 2 months of cryopreservation by the addition of 1M glycerol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C$, and $33\%,\;at\;-70^{\circ}C$. pollowing 2 months of cryopreservation by the addition of 1.5M DMSO of cryoprotectant, there were $27\%$ of fertilization rate at $-20^{\circ}C,\;an\;36\%\;and \;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1.5M glycerol of cryoprotectant, there were $34\% \;of\;fertilization\;rate\;at\;-20^{\circ}C, \;and\;31\%\;and\;31\%,\;respectively,\;at \;-40^{\circ}C\;and\;-70^{\circ}$. Following 2 months of cryopreservation by the addition of 1.5M methanol of cryoprotectant, there were $28\%$ of fertilization rate at $-20^{\circ}C,\;and\;29\%\;and\;28\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C.$ From 10 days and 15 days following fertilization at $13^{\circ}C\;and\;10^{\circ}C$, respectively, the mortality rate of fertilized ova was markedly increased. The middle piece of spermatozoa had two set of central doublets, nine set of outer coarse fibres, and mitochondrial sheath. Spermatozoa went through morphological changes during storage, e.g. winding of flagella, detachment of the nuclear envelope and the plasma membrane from the nucleus of the sperm head. There were $1\%$ abnormal spermatozoa in fresh sperm and about $15\%$ during storage.

• Journal of Chest Surgery
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• v.37 no.10
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• pp.817-826
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• 2004

Background: Several studies have demonstrated that conventional hypothermic cardiopulmonary bypass (CPB) causes cellular injury, abnormal responses in peripheral vascular beds and increased postoperative bleeding, whereas normothermic CPB provides protection of the hypothermic-induced effects and better cardiac recovery. The present study was prospectively performed to compare the effects of normothermic CPB to those of hypothermic CPB on the inflammatory and hematologic responses during cardiac surgery. Material and Method: Thirty-four adult patients scheduled for elective cardiac surgery were randomly assigned to hypothermic CPB (nasopharyngeal temperature $26～28^{\circ}C,$ n=17) or normothermic CPB (nasopharyngeal $temperature>35.5^{\circ}C,$ n=17) group. In both groups, cold $(4^{\circ}C)$ crystalloid cardioplegia was applied for myocardial protection. Blood samples were drawn from radial artery before (Pre-CPB), 10 minutes after starting (CPB-10) and immediately after ending (CPB-OFF) CPB. Total leukocyte and platelet counts, interleukin-6 (IL-6) level(expressed as percent to the baseline of Pre-CPB), D-dimer level, protein C and protein S activity were measured with the blood samples. The amount of bleeding for postoperative 24 hours and blood transfusion after operation were also assessed. All parameters were compared between the two groups. Result: The total leukocyte counts $(10,032\pm65/mm^3)$ and the increased ratio of IL-6 $(353\pm7.0%)$ at CPB-OFF in the normothermic group were higher than that $(7,254\pm48/mm^3$ and $298\pm7.3%)$ of the hypothermic group(p=0.02 and p=0.03). In the normothermic group, protein C activity $(32\pm3.8%)$ and protein S activity $(35\pm4.1%)$ at CPB-OFF were significantly lower than that $(45\pm4.3%$ and $51\pm3.8%)$ of the hypothermic group (p=0.04 and p=0.009). However, there were no differences in platelet counts and D-dimer concentration. In the normothermic group, the amount of bleeding for postoperative 24 hours $(850\pm23.2$ mL) and requirements for blood transfusion after operation such as packed cell $(1,402\pm20.5$ mL), fresh frozen plasma $(970\pm20.8$ mL) and platelet $(252\pm6.4$ mL) were higher than that $(530\pm21.5$ mL, $696\pm15.7$ mL, $603\pm18.2$ mL and $50\pm0.0$ mL) of the hypothermic group. Conclusion: These results indicate that normothermic CPB with cold crystalloid cardioplegia was associated with higher increase in inflammatory response, hemostatic abnormalities and postoperative bleeding problem than moderate hypothermic CPB.