This study was carried out to investigate levels of the components of microsomal mixed function oxidase (MFO) system and activities of the hepatic microsomal cytochrome P45O (P45O)-dependent monooxygenases of grass snake (Natrix tigrina Lateralis) and to compare with those of rat. The levels of P45O and cytochrome b$_{5}$, (b$_{5}$) of snake were much lower than those in rat. NADPH-cytochrome c reductase activity in the snake was also only 40% of that in the rat. Activities of 7-ethoxycoumarin 0-deethylase (ECOD) and benzphetamine N-demethylase (BPDM) of snake hepatic microsomes, when compared with those of rat, were markedly low. But, aryl hydrocarbon hydroxylase (AHH) and testosterone hydroxylase (TSH) activities were nearly the same or higher than those of the rat. Of the P45O-dependent TSHs measured, 7$\alpha$-hydroxylase activity was the highest in snake, whereas, 6$\beta$-hydroxylase activity was the highest in rat. However, stereoselectivity of the enzyme from the snake to C2 and C6 positions of testoste-rone was the same as rat. The result of radioimmunoassay (RIA) for the identification of five P45O isozymes with MAbs shows that relatively high content of ethanol-inducible P45O isozyme, CYP2El, exists in the rat, whereas MC-inducible P45O isozyme, CYP2A1/1A2, does in the snake. From the analyses of SDS-PAGE and RIA of partially pu-rified P45O, we suggest the possibility of the presence of a certain P45O isozyme(s) in hepatic microsomes of snake different from those of rat.
Type I allergic reaction and it's related clinical manifestations are known to occur by the effects of various chemical mediators. These chemical mediators are released from circulating basophils and tissue mast cells, which become 'sensitized' through the binding of antigens and antibodies of the IgE type to their cell surface receptors. Efforts to elucidate the mechanism of the release of these mediators, especially that of histamine, have been persued for years. The mechanism is not yet clarified at the present time. Recent reports of hyaluronidase, an enzyme known to be involved in the tissue inflammatory process, as possible participant in type I allergic reaction, initiated this study. Relationships between the hyaluronidase activity and histamine release from the sensitized rat peritoneal mast cells were investigated. Also anti-allergic agents, tranilast and disodium cromoglycate, along with known histamine releasers, morphine and compound 48/80, were used to observe the inhibitory and stimulatory effects of these substances on the hyaluronidase activity as well as histamine release from the rat mast cells. The results obtained are summarized as follows: 1) Hyaluronidase activity and histamine release from sensitiaed rat peritoneal mast cells started to increase on the 4th day of postsensitization. Hyaluronidase activity reached it's peak value on the 7th day of postsensitization and that of histamine release on the 14th day of postsensitization. 2) Hyaluronidase activity and histamine release from sensitized rat peritoneal mast cells, pre-treated with tranilast revealed significant decrease in comparison with those of non-treated cells. 3) Hyaluronidase activity and histamine release from sensitized rat peritoneal mast cells, pre-treated with tranilast, followed by morphine injection, revealed significant increase in comparison with those of tranilast treated cells. 4) In vitro study of hyaluronidase activity and histamine release from un-sensitized rat peritoneal mast cells, using morphine and compound 48/80 as activators, revealed significant increase compared to those of non-activator used cells. 5) In vitro study of hyaluronidase activity and histamine release from un-sensitized rat peritoneal mast cells, pre-treated with tranilast and disodium cromoglycate, using confound 48/80 and morphine as activators revealed significant decrease in comparison with those of tranilast and disodium cromoglycate treated cells. From above results, participation of enzyme hyaluronidase in the process of histamine release from sensitized rat pertioneal mast cells, could be suggested. It was also quite evident that the clinically used anti-allergic agents, tranilast and disodium cromoglycate, have significant inhibitory function on the hyaluronidase activity and histamine release from sensitized rat peritoneal mast cells, while morphine significantly increased the hyaluronidase activity and histamine release from sensitized rat peritoneal mast cells.
To see both $Sh{\grave{u}}nq{\grave{i}}daot{\acute{a}}nt{\bar{a}}ng$(dissipate phlegm and promote vital energy circulation) and $Hu{\grave{a}}y{\bar{u}}t{\bar{a}}ng$(blood circulation and disperse blood stasis) influencing on thrombosis, contusion-hyperemia, and hyperlipidemia, at first we measured the density of FDP, the quantity of fibrinogen, prothrombin time, and the number of platelet of rat taken thrombosis by endotoxin. Secondly we measured the increase-rate of "paw swelling", the number of platelet, the quantity of fibrinogen, and prothrombin time of rat taken contusion-hyperemia. And then we measured the quantity of total cholesterol in serum and of H.D.L-cholesterol and of triglyceride and of phospholipid and of ${\beta}-lipoprotein$, its weight, and the variation of the quantity of electrolyte of rat taken hyperlipidemia by the oral-injection of choleserol. As a result, we can conclude as follows : 1. Out of the test of thrombosis, we can recognize not only the noticeable increae of the number of platelet and the quantity of fibrinogen, but also the noticeable decrease of prothrombin time and the density of FDP in case of $Sh{\grave{u}}nq{\grave{i}}daot{\acute{a}}nt{\bar{a}}ng$-injected rat and $Hu{\grave{a}}y{\bar{u}}t{\bar{a}}ng$-injected rat. 2. Out of the test of contusion-hyperemia, we can recognize not only the noticeable increase of the number of platelet and the quantity of fibrinogen, but also the noticeable decrease of prothrombin time and "increase-rate of paw swelling" in case of $Sh{\grave{u}}nq{\grave{i}}daot{\acute{a}}nt{\bar{a}}ng$-injected rat and $Hu{\grave{a}}y{\bar{u}}t{\bar{a}}ng$-injected rat. 3. Out of the test of hyperlipidemia, at first we can recognize that test rat's weight increased as close as that of normal rat. And we can recognize the noticeable decrease of the triglyceride and phospholipid and ${\beta}-lipoprotein$." Also, in case of the variation of electrolyte we can recognize the decrease of calcium and potassium in $Sh{\grave{u}}nq{\grave{i}}daot{\acute{a}}nt{\bar{a}}ng$-injected rat, and of sodium and magnesium in $Hu{\grave{a}}y{\bar{u}}t{\bar{a}}ng$-injected rat. Thus, as the above-mentioned, in covering thrombosis, contusion-hypermia, and hyperlipidemia, the effect of $Sh{\grave{u}}nq{\grave{i}}daot{\acute{a}}nt{\bar{a}}ng$ and $Hu{\grave{a}}y{\bar{u}}t{\bar{a}}ng$ can be recognized. Granting that $Hu{\grave{a}}y{\bar{u}}t{\bar{a}}ng$ reveals its effectiveness in thrombosis and contusion-hyperemia, and $Sh{\grave{u}}nq{\grave{i}}daot{\acute{a}}nt{\bar{a}}ng$ in hyperlipidemia, it can be inferred that contusion-hyperemia is like "model of blood stasis form" as thrombosis and hyperlipidemia "phlegm-retention diseases form", and both phlegm-retention and blood stasis have correlation each other.
The present study was carried out to sex effectively mouse and rabbit embryos using rat H-Y antiserum and to improve the rate of rat producing H-Y antiserum with H-Y antibody. The H-Y antiserum was prepared from inbred SD strain female rat immunized by intrasplenic injection and subsequent intraperitioneal booster of testis cell of syngenic male newborn rat. the titer of antiserum was identified by in vitro cytotoxicity test of mouse embryos. The rabbit embryos exposed to the H-Y antiserum was classified to the developed (H-Y negative) or delayed (H-Y positive) group. The H-Y negative rabbit embryos were transferred to recipients and sex of offspring was examined. 1. When mouse embryos were exposed to the rat H-Y antisera, the ratio of embryos developed vs delayed was various. The H-Y antisera where the ratio of embryos developed vs delayed showed the range of 40~60% were recognized as having titer of H-Y antibody. 2. When the subsequent intraperitioneal boosters were followed after priming of intrasplenic injection of H-Y antigen, the rate of rat producting the H-Y antiserum with H-U antibody was 13, 27, 70 and 73% in control, 1st B, 2nd B and 3rd B, respectively. The rate in 2nd B and 3rd B was significantly(P<0.05) higher than that in control and 1st B. 3. When the rabbit morulae were exposed to the rat H-Y antiserum with H-Y antibody, the ratio of morulae developed versus delayed was 42:58% and it was close to the natural sex ratio 50:50%. It was confirmed that the rat H-Y antiserum was cross reactive to the rabbit morulae. 4. When the H-Y negative rabbit embryos were transferred to the recipients, the pregnancy rate was 50% and 83% of the newborns were females. In conclusion, the rat H-Y antiserum with high titer of H-Y antibody was able to be obtained from the female rat immunized by the intrasplenic injection followed by the second intrapent oneal booster of testis cells at a week interval. When the rabbit embryos negative to the rat H-Y antiserum were transferred, 83% of the newborns were females.
In order to examine if arsenic, one of environmental stresses, contributes to hypertension as one of cardiovas cular pathological factors, this study was perfarmed in vivo and in vitro, using intacted or pithed rats and aorta ring preparation, respectively. And also the relationship between expression of heat shock protein (HSP) 90 and vasoactives-induced contractile response was elucidated. To measure blood pressure, the carotid arterial pressure was recorded on physiograph(Grass Co. 79E) connected to strain gauge. On the other hand, contractile response of vascular ring preparation isolated from rat was determined in organ bath and was recorded on physiograph connected to isometric transducer. And HSP was detacted by Western blotting whole cell Iysis. Preganglionic nerve stimulation was increased by 26.0% in arterial pressure of rat treated with arsenic. Vascular contractile response was monitored and HSP were measured by Western blotting of whole Iysis prepared from samples exposed with 0, 0.5, 1, 2 and 4 mM of arsenic for 8 hours. The dose-vascular responses of potassium chloride were augmented by increasing dose of arsenic in the strips exposed to arsenic for 8 hours, and were not augmented for 1, 3, 5 hours. And the response of relaxation of rat aorta induced by histamine was not influenced by arsenic stress. The increase of HSP 90 expression in rat aorta was pronounced at 8 hours after 4 mM of arsenic treatment, but HSP 60 expression was not. Arsenic stress not only increased the expression of HSP 90 in the rat aorta, but also augmented contractions to potassium chloride. These results indicated that arsenic stress was sufficient to induce heat shock protein 90, resulting in increased vascular contractility in rat aorta.
Astrocytes generate free radicals including nitric oxide (NO) and reactive oxygen intermediates(ROI) which in turn play roles in the pathogenesis of degenerative diseases and sclerotic changes of the brain. This study was designed to evaluate the mechanism that free radicals contribute to the cytotoxicty of rat neonatal primary astrocytes. Treatment with NO donors alone including soldium nitroprusside(SNP), S-nitrosoglucathinoe (GSNO), and S-nitroso-n-acetylpenicillamine (SNAP) showed a little effect on the death of rat neonatal primary astrocytes, whereas SNP markedly induced the death of RAW 264.7 cells. ROI inculding H2O2 and O2 donor also slightly induced the death of rat primary astrocytes. However, 3-morpholinosydnonimine(SIN-1), a donor of peroxynitrite (ONOO), which is a reactive compound of NO with superoxide, significantly decreased the viability of rat primary astrocytes in a dose-dependent manner. Cells were retarded in outgrowth of viability of cellular processes with cell shrinkage and detachment from culture dishes. Hoechst staining demonstrated that SIN-1-induced cell death might be due to an apoptosis which was characterized by nuclear condensation and fragmentation. SIN-1-induced apoptosis was prevented by the pretreatment with superoxide dismutase (SOD) and catalase in rat primary astorocytes. Furthermore, prevention of the generation of reduced glutathione (GSH) by DL-buthionine-[S, R]-sulfoximine (BSO) aggravated the cytotoxic effects of SNP, benzene triol, and SIN-1 in rat primary astrocytes. Taken together, it is suggested that peroxynitrite may be a major effector of apoptosis and cellular antioxidant system is important for cell survival in rat prima교 astrocytes.
Journal of the Korean Society of Food Science and Nutrition
/
v.14
no.1
/
pp.13-22
/
1985
A study was undertaken to evaluate the nutritional quallity of protein from precooked seafoods. Procedures for evaluation included protein efficiency ratio(PER) using the rat, computed PER(C-PER) and discriminant computed PER(DC-PER) techniques. These procedures involve the determination of in vitro digestibility and amino acid composition of the sample prior to computation of C-PER and CD-PER value in laver was higher. For the oyster, the C-PER value was very close to the PER value obtained from the rat assay. The difference between DC-PER value and rat-PER or NPR was slightly lower than that between C-PER and rat-PER except oyster and laver. Seafood samples which posses a high in vitro protein digestibility may need the DC-PER procedure could offer more advantages in predicting the protein quality of seafood samples than the DC-PER procedure which showed poor in vitro digestibility.
Proceedings of the Korean Society of Applied Pharmacology
/
1997.04a
/
pp.94-94
/
1997
The objective of this study was to generate and characterize monoclonal antibodies against rat airway mucins, and therefore, should serve as a useful tool in studying the regulation of airway mucins using various in vivo rat models that are currently available. As an antigen, we used a high molecular mass mucin preparation purified from the spent media of rat tracheal surface epithelial cells in primary culture. Seven hybridomas were obtained which secrete monoclonal antibodies against the rat mucin among which mAbRT03 showed the highest immunoreactivity against the mucin based on ELISA. All of the antibodies secreted by these hybridomas recognized carbohydrate epitopes but not sialic acid residues since their immunoreactivity was completely abolished by treatment of the mucin with 20 mM periodate but not with neuraminidase. Further characterization of mAbRT03 showed that: (1) it belongs to the IgM type, (2) it binds to high molecular mass mucins based on both Western blot analysis and indirect immunoprecipitation, (3) it binds to the luminal side of tracheal epithelium as well as some goblet cells based on immunohistochemistry, and (4) it also recognizes in vive airway mucins from rats but not from human nor hamsters which have been purified from the airway lavage fluids. This is the first anti-rat airway mucin monoclonal antibody which has been developed against purified rat airway mucins. Therefore, mAbRT03 should be able to serve as an invaluable tool in studying the regulation of airway mucins using various intact rat models that are currently available.
Objective : The authors evaluated the accuracies and ease of use of several commonly used microanastomosis training models (synthetic tube, chicken wing, and living rat model). Methods : A survey was conducted among neurosurgeons and neurosurgery residents at a workshop held in 2009 at the authors' institute. Questions addressed model accuracy (similarity to real vessels and actual procedures) and practicality (availability of materials and ease of application in daily practice). Answers to each question were rated using a 5-point scale. Participants were also asked what types of training methods they would chose to improve their skills and to introduce the topic to other neurosurgeons or neurosurgery residents. Results : Of the 24 participants, 20 (83.3%) responded to the survey. The living rat model was favored for model accuracy (p<0.001; synthetic tube $-0.95{\pm}0.686$, chicken wing, $0.15{\pm}0.587$, and rat, $1.75{\pm}0.444$) and the chicken wing model for practicality (p<0.001; synthetic tube $-1.55{\pm}0.605$, chicken wing, $1.80{\pm}0.523$, and rat,$1.30{\pm}0.923$). All (100%) chose the living rat model for improving their skills, and for introducing the subject to other neurosurgeons or neurosurgery residents, the chicken wing and living rat models were selected by 18 (90%) and 20 (100%), respectively. Conclusion : Of 3 methods examined, the chicken wing model was found to be the most practical, but the living rat model was found to represent reality the best. We recommend the chicken wing model to train surgeons who have mastered basic techniques, and the living rat model for experienced surgeons to maintain skill levels.
These expriments were carried out to investigate existence of H-Y antibody in the rat serum immunized against H-Y antigen from rat spleen cells and effect of H-Y antiserum on development of mouse male embryos. The results obtained were summerized as follows : 1. When mouse embryos were cultured for 48∼72 hrs in the Ham's F10 containing 16% of FBS(fetal bovine serum) or RNS(rat normal serum), percentages of embryos developed from 2, 4, 8 and 16-cell embryo to morulae were 20, 27, 94 and 100%, respectively, in FBS and 8, 7, 94 and 100%, respectively, in RNS. Eight to 16-cell embryos showed no difference in development rate between FBS adn RNS. 2. When 8∼16-cell mouse embryos were cultured for 24∼48 hrs in the Ham's F10 containing FBS, RNS+GPC(guinea pig complement) and RAS(rat antiserum)+GPC, proportions of embryos developed to the expanded blastocyst stage were 100, 82.4 and 52.1∼53.6%(ave.52.9), respectively, so that it was suggested that rat antiserum suppressed development of male embryos. 3. When 8∼16-cell mouse embryos were cultured for 24∼48 hrs in the Ham's F10 containing FBS, RNS, RNS+GPC and RAS+GPC, proportions of embryos developed to the expanded blastocyst stage were 94.5, 90.9, 82.3 and 47%, respectively, and the embryos developed in the medium containing RAS+GPC seemed to be female. These results indicated that the antisera prepared through immunized against H-Y antigen from rat spleen cell, possessed H-Y antibody which supressed development of male embryos.
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