In this study, poikilocyte rate and histopathological observation of flounder, Paralichthys olivaceus, treated with formalin, neutral-formalin(0, 50, 100, 150, 200, 300 ppm) at two different temperatures (15 and $25^{\circ}C$) were conducted to determine the time need for recovery. In all test chamber, formalin was more toxic than neutral-formalin and moderate lesions were not observed at low concentration of formalin and neutral-formalin(50~100 ppm). As expectedly, time need for recovery of gill and kidney tissues was lengthening as increasing chemical concentration and temperature. Treatments caused edema, winding of secondary gill lamella, seperation of epithelial layer and thrombosis of secondary gill lamella in the gill; edema, hydropic and hyaline droplet degeneration in the kidney. Representatively, recovery period of fish gill that had been exposed to 300 ppm formalin and neutral-formalin was about 120 and 72 hr($25^{\circ}C$). 72 and 48 hr($15^{\circ}C$). Recovery period of kidney was about 72 and 48 hr($25^{\circ}C$), 48 and 48 hr($15^{\circ}C$) respectively. Maximum value of poikilocyte rate(27.84%) was shown in formalin 300 ppm treated fish at $25^{\circ}C$. Tendency of decreasing poikilocyte rate was similiar to recovery of tissues in treated fish.
For the preparation of $^{99m}Tc$-labeled RBC, $10{\sim}20\;{\mu}g/kg$ of Stannous(II) chloride and $10{\sim}40\;min$ of preparation was used. For finding out the effect of contrast agent, the blood samples were collected in three days, seven days, and 1 months after the diagnostic procedure. In the normal volunteer, the concentration of reducing agent and preparation time did not effect on the radiochemical yield. But in the patients, 10 mg of Stannous(II) chloride and 60 min incubation times was shown high radiochemical yield. Contrast agent has a significant effect on the radiochemical yield. Although the blood samples which were collected after seven days of diagnostic procedure did not effect on the radiochemical yield of $^{99m}Tc$-labeled RBC, but the radiochemical yield of $^{99m}Tc$-labeled RBC which was prepared with a sample of high concentration of contrast agent in blood led to low radiochemical yield. For these samples, the modified method showed high radiochemical yield than previous in vivo preparation method. The recommended method is followed. Blood collecting was performed at 30 minutes after injection of reducing agent, and it is centrifuged for removal of plasma. After addition of $^{99m}TcO^-_4$, sample reservoir was rotated. After addition of normal saline, and it is centrifuged for separation of saline. Then $^{99m}Tc$-labeled RBC was obtained after removal of saline.
In cardiovascular disease the flow adaptation of erythrocytes can be affected by reduced shear stresses and metabolic influences on red cell fluidity as a consequence of tissue hypoxia. In addition there are indications that risk factors of cardiovascular diseases are able to decrease the intrinsic red cell deformability. Erythrocyte deformability was studied by the filtration technique of Reid et al. to investigate the relationship between blood pressure chances and erythrocyte deformability. In this experiment normotensive rats, spontaneously and DOCA-salt treated hypertensive rats were used. Erythrocyte deformability was significantly reduced by blood pressure elevation in hypertensive rats but was not fully recovered by normalization of blood pressure after antihypertensive drug treatment. Therefore other factors than blood pressure may be involved in erythrocyte deformability reduction during blood pressure elevation.
Histopathological changes and methemoglobinemia of the eels (Anguilla japonica) reared at six culture farms in the vicinity of Y$\check{o}$su city were investigated under the conditions of $22.0{\sim}29^{\circ}C$, 4.33~7.33ppm of D.O., pH 6.1~8.2 and 0.03~10.1ppm of nitrite from December, 1989 to June, 1990. All of the eels showed no any abnormal state in action, color and blood, but the eels reared in pH 6.0 had planty of mucus and stickiness on their body surface. Methemoglobin levels were 1.03~9.86% of total erythrocyes in Dec. and 5.05~25.69% in June, and poikilocyte levels were 5.8~30.17% in Dec. and 8.67~74.45% in June. These results indicate the fact that the water management of the culture farms was worse in summer than in winter, even though not good in winter. Hypertrophy was found in the gill, hepatic and kidney tissue of the eels in all the culture farms. In March and June, Fish of all culture farms appeared the detachment of gill lamella epithelium cells, pyknosis and sinusoids of nucleus in the hepatic tissue and hemosiderosis in the kidney tissue. The trend of increase in methemoglobin levels appeared in the fish of culture farms with high nitrite concentration, which didn't appear equally in all the culture farms. However, in the culture farms of pH 6.0, methemoglobin and poikilocyte levels were increased independently of concentration of nitrite, and degeneration in the tissue of the gill, liver and kidney was also appeared.
Seo, Han-Kyung;Kim, Min-Woo;Lim, Seok-Tae;Sohn, Myung-Hee
The Korean Journal of Nuclear Medicine
/
v.38
no.4
/
pp.300-305
/
2004
Purpose: We underwent this study to evaluate the factors which influence labeling efficiency when modified in vivo erythrocyte labeling technique was used. Materials and methods: Thirty healthy volunteers (M:F=19:11, age:$25{\pm}2$ yrs) were enrolled in this study. Totally, two hundred ten samples were obtained from them. The 1 mg of stannous pyrophosphate was injected intravenously at the beginning of labeling. After suitable tinning time (5 min, 20 min, 35 min) passed by, blood (5 mL, 3 mL or 1 mL) was withdrawn into 10 mL syringe previously containing Tc-99m (740 MBq) and anticoagulant (heparin, ACD or CPDA) through 19-gauged scalp needle. The generator ingrowth time of Tc-99m was within 24 hrs in each case. The blood samples were placed on rotating invertor during incubation (10 min, 25 min, 40 min) but some of them were not. Immediately after the conclusion of incubation, the labeled blood specimens to analyze were centrifuged. and then %Unbound Tc-99m was calculated. Statical analysis was used paired T-test and one way ANOVA with SPSS 10.0. Results: The binding efficiency at 1 mL of blood volume was $73{\pm}32%,\;91{\pm}10%$ at 3 mL and $96{\pm}7%$ at 5 mL (p<0.01). The binding efficiency at 5 min of tinning time was $45{\pm}23%,\;98{\pm}6%$, at 20 min and $97{\pm}8%$ at 35 min (p<0.001). The binding efficiency at 10 min of incubation time was $96{\pm}7%,\;95{\pm}12%$ at 25 min and $98{\pm}3%$ at 40 min (p>0.05). The binding efficiency in case of using rotating invertor was $96{\pm}7%$ and the binding efficiency in case of not using it was $87{\pm}18%$ (p>0.05). There was no significant difference between them. In binding efficiency according to kinds of anticoagulants, ACD was $98{\pm}4%$, CPDA was $97{\pm}6%$ and heparin was $89{\pm}20%$ (p<0.001). Conclusion: When modified in vivo erythrocyte labeling technique is used with Tc-99m, the methods to obtain the highest labeling efficiency are as follow. The withdrawing blood volume should be over 3 mL, tinning time should be kept between 20 min and 35 min, and incubation time should be kept between 10 min and 40 min. ACD or CPDA have to be used as a anticoagulant except heparin and the blood samples should be placed on rotating invertor during incubation.
Background: Retrograde autologous priming (RAP) is known to be useful in decreasing the need of transfusions in cardiac surgery because it prevents excessive hemodilution due to the crystalloid priming of cardiopulmonary bypass circuit. However, there are also negative side effects in terms of blood conservation. We analyzed the intraoperative blood-conserving effect of RAP and also investigated the efficacy of autotransfusion and ultrafiltration as a supplemental method for RAP. Material and Method: From January 2005 to December 2007, 117 patients who underwent isolated coronary artery bypass operations using cardiopulmonary bypass (CPB) were enrolled. Mean age was 63.9$\pm$9.1 years (range 36$\sim$83 years) and 34 patients were female. There were 62 patients in the RAP group and 55 patients in he control group. Intraoperative autotransfusion was performed via the arterial line. RAP was done just before initiating CPB using retrograde drainage of the crystalloid priming solution. Both conventional (CUF) and modified (MUF) ultrafiltrations were done during and after CPB, respectively. The transfusion threshold was less than 20% in hematocrit. Result: Autotransfusions were done in 79 patients (67.5%) and the average amount was 142.5$\pm$65.4 mL (range 30$\sim$320 mL). Homologous red blood cell (RBC) transfusion was done in 47 patients (40.2%) and mean amount of transfused RBC was 404.3$\pm$222.6 mL. Risk factors for transfusions were body surface area (OR 0.01, 95% CI 0.00 $\sim$ 0.63, p=0.030) and cardiopulmonary bypass time (OR 1.04, 95% CI 1.01 $\sim$ 1.08, p=0.019). RAP was not effective in terms of the rate of transfusion (34.5% vs 45.2%, p=0.24). However, the amount of transfused RBC was significantly decreased (526.3$\pm$242.3ml vs 321.4$\pm$166.3 mL, p=0.001). Autotransfusion and ultrafiltration revealed additive and cumulative effects decreasing transfusion amount (one; 600.0$\pm$231.0 mL, two; 533.3$\pm$264.6 mL, three; 346.7$\pm$176.7 mL, four; 300.0$\pm$146.1 mL, p=0.002). Conclusion: Even though RAP did not appear to be effective in terms of the number of patients receiving intraoperative RBC transfusions, it could conserve blood in terms of the amount transfused and with the additive effects of autotransfusion and ultrafiltration. If we want to maximize the blood conserving effect of RAP, more aggressive control will be necessary - such as high threshold of transfusion trigger or strict regulation of crystalloid infusion, and so forth.
Cardiopulmonary bypass in children is associated with capillary leak which results in an increase in total body water after open heart surgery The purpose of these studies was to assess the cardiopulmonary effects of modified ultrafiltration after pediatric open heart surgery Study h: Twenty-six consecutive children aged 0.1 ~ 10 years(median 7 months) underwent cardiac operation inc rporating modified ultrafiltration. After completion of cardiopulmonary bypass, modified ultrafiltration was commenced at the flow rate of 100~ 15011min for 3 ~ 14 min. After modified ultrafiltration, elevation of hematocrit(28.3% $\pm$ 3.6% vs. 33.8olo $\pm$ 4.Ooloi p < 0.001), increased systolic 1)loots Pressure(66.7 $\pm$ 11.2mmHg vs. 76.2$\pm$ 11.BmmHg, p < 0.02), and decreased central venous pressure(7.8 $\pm$ 3.7mmHg vs. 6.9$\pm$ 2.gmmHg, p<0.001) were observed. Study B: Twenty-six children who underwent cardiac operation with the diagnosis of VSD under 2 years were assigned to control(n= 14) or modified ultrafiltration(n= 12). Peak inspiratory pressure checked immediately after operation was significantly lower in modified ultrafiltration group than in control group(20.0$\pm$ 2.4 cmH20 vs.22.4$\pm$ 2.3cmH20, p < 0.03). Modified ultrafiltration after cardiopillmonary bypass in children improves early homodynamics and pulmonary mechanics, and represents an excellent option for perioperative managemen of accumulation of fluid in the tissues. We will continually employ the modified ultrafiltration technique in pediatric cardiac operations.
Journal of the Korea Academia-Industrial cooperation Society
/
v.20
no.8
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pp.417-424
/
2019
Despite its superior ability to show distinct cellular morphology and for long-term storage, conventional tissue fixation by formalin has many drawback, including slower fixation, the exposure to harmful chemicals and extensive protein modification. Herein, we assessed the effects of rapid microwave-assisted tissue fixation on histological examination and on protein integrity by comparing these microwave irradiation fixated tissues with the formalin-fixed tissues. One of the paired mouse tissues (liver and kidney) was fixed in formalin and the other was fixed by using microwave irradiation in phosphate buffered saline. Each slide from the paraffin-embedded tissues was examined by H & E staining for the adequacy of fixation and by immunohistochemical staining for antigenicity in a blinded fashion. Evaluation of protein recovery and the protein quality from the fixed tissues were analyzed by the BCA method and Western blotting, respectively. The results from H & E staining and immunohistochemical staining showed that the sections obtained from microwave-fixed tissues under our experimental conditions were comparable to those of the formalin-fixed tissues except for the integrity of RBCs. Furthermore, proteins were effectively extracted from the microwave-fixed tissues with acceptable preservation of the proteins' quality. Taken together, this microwave-assisted tissue processing yields a quick fixation and better protein recovery in higher amounts, as well as the adequacy of fixation and the antigenicity being comparable to formalin-fixed tissues, and this all suggests that this new fixation technique can be applied in an environment where rapid tissue fixation is required.
Objectives : The aim of this study was to assess the relationship between metabolic syndrome and erythrocyte deform ability in acute stroke patients. Methods : Among 88 of the recruited patients, 52 were diagnosed as metabolic syndrome. We assessed their general characteristics, risk factors. We compared the assessed variables between metabolic syndrome and control group. We analyzed the relationship between metabolic syndrome and erythrocyte deform ability. We analyzed relationship between cardiovascular risk factors and erythrocyte deformability. Results : The general characteristics waist and hip circumference, waist/hip ratio were higher in metabolic syndrome group. The metabolic syndrome group was also diagnosed with hypertension, DM, and hyperlipidemia more often than the control group. The blood test metabolic syndrome group showed higher triglycerides, total lipids, fasting blood sugar, and 2 hours postprandial plasma glucose level and lower HDL-cholesterol than the control group. There were more patients diagnosed with Dampness-Phlegm in the metabolic syndrome group. There were more patients showing lower erythrocyte deform ability in the metabolic syndrome group. The plasma homocysteine level was negatively correlated with erythrocyte deform ability. Conclusion : The results reconfirmed that the risk factors are more in metabolic syndrome group. The results indicated that metabolic syndrome lead to a lower erythrocyte deform ability in small vessel disease stroke patients. The Plasma homocysteine level was negatively correlated with erythrocyte deform ability.
Objectives : Coldness of hands and feet has been regarded to be associated with blood circulation. This study was performed to assess the correlation between erythrocyte deformability and coldness of hands and feet. Methods : 145 patients who had taken an erythrocyte deformability test were selected for this study among hospitalized patients within 4 weeks after their cerebral infarction at the Internal Medical Department at Kyunghee Oriental Medical Center from August 2008 to August 2009. We divided the selected patients into two groups, with and without of coldness of hands and feet groups. The two groups were compared on risk factors for atherosclerosis and examined for variables including erythrocyte deformability. Results : 1. On demographic variables of the patients, only body mass index was significantly lower in the coldness of hands and feet group than the control group. There was no significant difference of the rate of hypertension, diabetes mellitus, hyperlipidemia, ischemic heart disease, smoking, drinking and carotid artery stenosis between the two groups. 2. According to blood test, the coldness of hands and feet group showed significantly lower erythrocyte deformability index than the control group. 3. Body mass index and erythrocyte deformability index showed a close relationship with coldness of hands and feet in multivariate analysis. Conclusions : The correlation between the erythrocyte deformability and coldness of hands and feet in cerebral infarction patients were decreased erythrocyte deformability participated in the mechanism of coldness of hands and feet.
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