It is widely known that pulsed magnetic field (PMF) is very useful tool to manipulate chemical and physiological processes in human body. The purpose of our study is to observe dynamics of rouleaux patterns of red blood cells (RBC) under PMF. The aggregation of RBCs or rouleaux formation is caused by fibrinogen in blood plasma. The maximum magnetic field intensity is 0.27 T and pulse time of 0.102 msec and pulse repetition rate was 1 Hz. PMF stimulus was applied to the palm of left hand for 5, 10, 15 and 20 min. Live blood analysis was used in vitro in order to quantitatively estimate the velocity of RBC exposed to PMF stimulus. The velocity of stacked-RBC of 10 minute PMF stimulus was increased up to $8{\times}10^{-4}m/sec$, but it decreased rapidly as the time passed. The results of present study have adduced that PMF stimulus on hand provide the improvement of RBC rouleaux formation, increase of RBC's moving velocity as well as low blood viscosity.
Among the environmental pollutants, cadmium and lead compounds may impair human health. These compounds may inhibit the biological metabolic function of human body and may furthermore cause the disease directly or indirectly. This study was undertaken to investigate the effects of the immune response by intoxication of cadmium chloride and lead acetate. Cadmium chloride (8.8mg/kg, in saline 10ml) and lead acetate (15mg/kg, in saline 10ml) were administered by intraperitoneal injection. After 3 weeks, the rats were intoxicated with the above chemicals and immunized with sheep RBC. After 4 weeks, the immune response of rat spleen cells was measured by the Jerne's technique. The results were obtained as follows; 1. There was no change in leukocyte counts by the intoxication of cadmium chloride or lead acetate. 2. Cadmium chloride or lead acetate reduced hemoglobin contents for most intoxicated and immunized groups. 3. Hematocrits were decreased by the intoxication of cadmium chloride or lead acetate significantly. 4. It was determined that total protein, A/G (Albumin/Globulin), ${\alpha}-,\;{\beta}-\;and\;{\gamma}$-globulins in rat serum were not changed. 5. Intoxication by cadmium chloride or lead acetate reduced the number of hemolytic plaque to the sheep RBC in rat spleen cells. Therefore, antibody producing of rat spleen cells was suppressed by the intoxication of cadmium chloride and lead acetate.
This study was conducted to evaluate the effects of duck-meat consumption on adult disease risk factors, including body compositions, hematological variables, and serum metabolic and lipid profiles in adult human males. To obtain results, 20 adult males aged 20 to 25 were subjected to a diet of 600 g/day of duck-meat for 4 wk, after which body composition, hematological variables, and serum metabolic and lipid profiles were investigated to determine if there was a relationship between duck-meat consumption and adult disease risk factors. The results revealed that high amounts of duck-meat intake did not negatively alter body indices such as body weight, fat mass, body mass index (BMI), % body fat or waste-to-hip ratio (WHR). Furthermore, there were statistically insignificant changes in the number of blood cells, although this number did increase significantly following intake of duck-meat. Moreover, general decreases in serum metabolic parameters were observed, but none of these changes were significant with the exception of the concentration of blood urea nitrogen (BUN). The serum concentrations of LDL-cholesterol also showed a statistically significant decrease by 5.86%. Therefore, this study suggests that the ingestion of duck-meat not only significantly increased the RBC count but also decreased BUN and LDL-cholesterol concentrations in adult males.
Among the environmental pollutants, parathion, chloroform and 2, 6-dichlorophenol may impair human health; they may inhibit or reduce the metabolic function of human body and may furthermore cause diseases directly or indirectly. This study was undertaken to investigate the effects on the immune response by intoxication of parathion, chloroform or 2, 6-dichlorophenol. Parathion(1.3mg/kg, olive oil 10ml), chloroform (100mg/kg, olive oil 10ml) were administered via intraperitoneal injection to rats. And 2, 6-dichlorophenol (13mg/kg, olive oil 10ml) was administered via oral injection. After 3 weeks, the rats were intoxicated with the above chemicals and immunized with sheep RBC. After 4 weeks the immune response of rat spleen cells was measured by the Jerne's technique. The results were obtained as follows. 1. There was no change of leukocyte counts by the intoxication of parathion, chloroform and 2, 6-dichlorophenol. 2. Parathion, chloroform and 2, 6-dichlorophenol reduced hemoglobin contents for most intoxicated and immunized groups. 3. Hematocrits were decreased by the intoxication of parathion, chloroform or 2, 6-dichlorophenol significantly. 4. It was determined that total protein, A/G (albumin/globulin), .alpha.-, .betha.-and .gamma.-globulins in rat serum were not changed. 5. Intoxication by parathion, chloroform or 2, 6-dichorophenol reduced the number of hemolytic plaque to the sheep RBC in rat spleen cells. Therefore, the capacity of erythrocyte production and the immune response of rat spleen cells were decreased by the intoxication of parathion, chloroform, or 2, 6-dichlorophenol.
Membranes obtained from the normal human RBC population were separated by continuous sucrose density gradient centrifugation and the density-fractionated membranes were then examined for changes in molecular markers. This study focuses on changes of (i) the membrane protein profile, (ii) differences in membrane-associsted enzyme activities, and (iii) the amount of autologous IgG bound. The following observations were made: (i) ratios for band 4. la over the sum of bands (4. la + 4.Ib) ranged from 0.58 to 0.79 for membranes of lowest density; (ii) significant changes in bound glyceraldehyde-3-phosphate dehydrogenase and acetvlcholinesterase activities were found; (iii) the amounts of autolosous IgG's attached to the red blood cells was highest in the membrane fraction of lowest density.
Proceedings of the Korean Society of Embryo Transfer Conference
/
2002.11a
/
pp.97-97
/
2002
Human eryhropoietin (EPO) is acidic glycoprotein hormone that plays key role in hematopoiesis by facilitating differentiation of erythrocyte and formation of hemoglobin (Hb) and is used for the treatment of anemia. Human EPO is consist of 166 amino acids which is modified by three N-glycosylations (24, 38, 83) and single O-glycosylation (126). N-glycosylation is reported to be related to the cellular secretion and activity of EPO. In this study, we examined effects of mutagenesis in glycosylation site of recombinat hEPO for the cellular secretion during production from cultured CHO cell. We produced rhEpo which was cloned by PCR from human liver cDNA (TaKaRa) in cultured CHO cell. Using supernatant of the culture, ELISA assay and western analysis were performed. To estimate biological activity, 20IU of rhuEpo was subcutaneously injected into four ICR mice. After 8 days, HCT level was increased average 13 per cent, RBC was increased ca. 2${\times}$10$\^$6//${\mu}\ell$. In disease model Rat (anemia c-kit, WSRC-WS/WS), HCT was increased ca. 12%, RBC was increased ca. 1.6${\times}$10$\^$6//${\mu}\ell$. These results suggests that rhEpo we produced has biological activity. To remove glycosylation site by substituting 24, 38, 83, and 126th asparagine (or serine) with glutamic acid, overlapping -extension site-directed mutagenesis was performed. To add novel glycosylation sites, 69, 105th leucine was mutated to asparagine. Mutant EPO construct was transfected into CHO cell. Supernatant of the cell culture was analyzed using ELISA assay with monoclonal anti-EPO antibody (Medac, Germany). Since, several reports for mutagenesis of glycosylation sites showed case-by-case results, we examined both transient expression and stable expression. Addition of novel glycosylation sites resulted no secretion while deletion mutants had little effect except some double deletion mutants (24/83 and 38/83) and triple mutant. We suggest that not single but combination of glycosyl group affect secretion of EPO.
Attempts were to produce porcine leukocyte interferon(PorLeIF) and porcine immune interferon (PorIIF) in the culture of porcine leukocytes. The interferons produced were tested for antiviral activity against vesicular stomatitis virus on poreine-derived PK(15) cells, human-derived FL cells, and Korean native black goat-derived BGK cells. The results were summarized as follws: 1. In the isolation of porcine leukocytes, the mean isolation rate by the buffy coat separation method (28.7%) was higher than that by the hydroxyethyl starch-RBC sedimentation method (9.2%). 2. When NDV(BI)-induced PorLeIFs were assyed on PK(15) cells and FL cells, the mean titers were 129 IU/ml and 72 IU/ml respectively, being 55.8% of the activity in homologous species system expressed in heterologous system. 3. The activities of PHA P-induced PorIIFs were 197 IU/ml on PK (15) cells and no activity on human FL cells. The mean antiviral activity of PorIIF was 1.5 times that of PorLeIF in PK (15) cells. 4. The cytopathic effect of vesicular stomatitis virus was observed in BGK cells derived from Korean native black goat kidney permitting interferon assay on the cells. While the cross-species antiviral activity of reference human ${\alpha},\;{\beta}-interferon$ was observed on the cells, PorLeIF and PorIIF did not show any activity.
The relative state of human iron storage may be ascertained more reliably through determination of the serum iron, iron binding capacity, transferrin saturation and absorption of radioactive iron in conjunction with studies of red cell morphology than from the study of red cell morphology alone. Recent investigations have shown that there is an increase in red cell protoporphyrin concentration in iron deficiency anemia. The significance of the red cell protoporphyrin has been discussed greatly during the years since its discovery. Two of the main factors which appear to influence the amaunt of protoporphyrin are increased erythropoiesis and factors interfering with the utilization of iron in the synthesis of hemoglobin, and iron deficiency. Recently Heller et al. have described a simplified method for blood protoporphyrin assay and this technique could be used assess nutritional iron status, wherein even minor insufficiencies are detectable as increased protoporphyrin concentrations. Based on the evaluation of the relationship between nutritional iron status and red cell protoporphyrin as an index suitable for the detection of the iron deficiency is described in this paper. RESULTS 1. Hemoglobin Concentrations and Anthropometric Measurements. The mean and standard deviations of the various anthropometric measurements of different age and sex groups are shown in table 1. There measurements have been compared with the Korean Standard. In the absence of local standards for arm circumference and skin-fold thickness over triceps, they have been compared with the standard from Jelliffe. Table 2,3, and 4 give anthropometric measurements and frequency (%) of anemia in children surveyed. The mean height of the children studid was 10 to 20 percent; below the Korean Standard. The distribution of height below 80 percent of the Standard was 21.2 percent, however, among anemic group this percentage was 27.7 percent. In general, the mean weight of the children was 10 to 15 percent below the Korean Standard. The percentage of children with weight less than 80 percent of the Standard was about 35 percent. But in the anemic group of the children, this percentage was 44 percent. The mean arm circumference was about 15 percent lower than the Jelliffe's standard. 61.2 percent of the children had values of arm circumference below 80 percent of the standard. Children with low hemoglobin levels, this percentage was 80 percent. The mean skinfold thickness over the triceps of the children studied was about 25 Percent lower than the Jelliffe's standard and 61.2 percent of the children had the value less than 80 percent of the standard. Among anemic children, this percentage was 70.8%. As may be seen from table 5, the mean hemoglobin concentration of the total group was 11.3g/100ml. Hemoglobin concentration was less than 11.0g/100ml. in 65(36.5%) of the 178 children. The degree of anemia in most of these children was mild with a hemoglobin level of less than 8.0g/100ml. found in only one child. In general, the prevalence of anemia was high in female children than male and decreased its frequency with increasing age. Relatively close relationship was observed between hemoglobin level and anthrophometric measurements especially high between arm circumference and skinfold thickness and hemoglobin but very low in height and low in weight and hemoglobin level, estimated by chi-square value. II. Serum iron, Transferrin saturation (1) Serum iron, and transferrin saturation Serum iron, transferrin saturation and red cell protoporphyrin concentrations were estimated in sub-sample of 84 children from 1 to 6 years and 24 older children between 7 and 13 years of age. The findings are presented in table 6. The mean serum iron concentration of the total group was 59ug/100ml. However, the level incrased with age from 36.6ug/100ml. (1-3years) to 80.8ug/100ml. (7-13 years). 60 percent of these children had a serum iron level less than 50ug/10ml. in the 1-3 years age group and 31.4 percent for 4-6 years group. These contrast with the finding of 12.5 percent anemic children in the 7-13 years age group. The mean transferrin saturation for the total group was 18.1 percent and frequency of anemia by transferrin saturation was observed same pattern as serum iron concentration. (2) Red cell protoporphyrin concentrations. (a) Red cell protoporphrin levels of children: Red cell protoporphyrin and other biochemical data are shown in table 4. The mean concentration in red cell of all children was fround 46.3ug/100ml. RBC. and differences with age groups were observed; in the age group 1-3 years, the mean concentration was $59.5{\pm}32.14$ ug/100ml. RBC; 4-6 years $44.1{\pm}22.57$ ug/100ml. RBC. and 7-13 years, $39.0{\pm}13.56$ ug/100ml. RBC. (b) Normal protoporphyrin values in adults: It was observed that in 10 normal adult males studied here the level of protoporphyrin in red cell ranged from 18 to 54 ug/100ml. RBC. and the mean concentration was $47.5{\sim}14.47$ ug/100ml. RBC. Other biochemical determination made on the same subjects are presented in table 8. (c) Red tell protoporphyrin concentration of occupational blood donors: The results of analyses for red cell protoporphyrin as well as serum iron, transferrin saturation and hemoglobin in the 76 blood donors are presented in table 7 and 8. In this experiment, donors were selected at random, however, most of them bled repeatedly because of poor economic situation, I doubt. Table 9 shows the distribution of red cell protoporphyrin concentration and hemoglobin concentration of occupational donors. The mean hemoglobin value for the total was 11.9 g/100 ml. When iron deficiency anemia is defined as a transferrin saturation below 15%, prevalence of anemia was 47.4 percent and the mean serum iron was 27.1ug/100ml. and red cell protoporphyrin, 168.3ug/100ml. RBC. However, mean serum iron and protoporphyrin concentration of above 15% transferrin saturation were 11.6 ug/100 ml. and 58.8 ug/100 ml. RBC. respectively. The mean Protoporphyrin concentration of non-anemic (above 15% transferrin saturation) donors was slightly higher than the results of normal adult males.
Effects of oral taurine supplementation (6g/day)for 2-4 weeks on activities of red blood cell(RBC)total superoxide dismutase (SOD) and plasma glutathione peroxidase(GSH-Px) and the level of malondialdehyde(MDA) were evaluated in healthy female adults (23.6$\pm$0.3 years old). Compared to the value for 0 week plasma GSH-Px activity of the subjects was significantly lower after 2 weeks of taurine supplementation(p<0.05) and recovered to the value similar to 0 week after 4 weeks of taurine supplementation. RBC total SOD activity tended to be decreased after 2 weeks of taurine supplmentation compared to the values for 0 week although the difference between the means of the two group was not statistically significant. Plasma MDA level was not significantly decreased by taurine supplementation most probably due to the fact that the subjects participated in the present study were healthy and their antioxidant defense system had been in the 'normal' range. Plasma MDA concentration was negatively correlated with plasma taurine concentration(r=-0.2003m p<0.05) but tended to be positively correlated with plasma cholesterol concentration(r=0.2465, p=0.0645) as expected Plasma GSH-Px activity was positively associated with the percentage of 22:0 (r=0.2892, p<0.05) or 20:4w6(r=0.2939, p<0.05). On the other hand plasma MDA concentration was positively correlated with the percentage of 20:5w3 in plasma total lipids(r=0.2635 p<0.05) and negatively correlated with $\Delta$5 desaturation index of w6 fatty acids(20:3w6⇒20:4w6) in plamsa total lipids(r=-0.2714, p<0.05) as well as in phospholipids(r=-0.2864, p<0.05). From these results protective effect of taurine supplementation against lipid peroxidation and antioxidant defense system in humans appears to be minimal when the subjects are in a relatively healthy state. Further studies concerning the antioxidant efficacy of taurine should be conducted in human subjects under various disease states related to oxidative stress such as diabetes and artheroxclerosis.
The present study were performed to analysis the hematocrit and the red blood cells content into the blood plasma of the transgenic pigs harboring recombinent human erythropoietin gene (rhEPO). Mouse whey acidic protein (mWAP) linked to rhEPO gene was microinjected into pronuclei of porcine one-cell zygotes. After delivered of offspring, PCR analyses identified one mWAP-rhEPO transgenic founder offspring(F$_{0}$). The first generation of transgenic pig (F$_{0}$) harboring mWAP-hEPO appeared to be a male, and the second generation (F$_1$) pigs were made by natural mating of F$_{0}$ with domestic swine, and male and female transgenic pigs (F$_1$) were identified by PCR. The blood samples from transgenic and normal pigs were collected for 50 days during lactation and were counted the red blood cell (RBC) numbers and Hematocrit (HCT) content into the blood. The transgenic pigs expressing rhEPO in their blood gave rise to higher RBC numbers and HCT contents than control animals. rhEPO was secreted both in the blood and milk of genetically engineered pigs harboring rhEPO gene. Therefore, this study provides a model regarding the production of transgenic pig carrying hEPO transgene for biomedical research.earch.
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