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Glucose-6-Phosphate dehydrogenase (G6PD) is an enzyme present in the cytoplasm of all cells in the body and protects red blood cells from certain attacks. The anomaly is genetically inherited in a recessive manner, linked to the X chromosome. Currently, an estimated 420 million individuals lack G6PD worldwide and its incidence varies between 1-25% in some African populations. This was an analytical cross-sectional study with prospective data collection that took place from May to November 2018, i.e. six (6) months. Our study included 108 blood donors, 88 men and 20 women. The samples were stored at 2-8 ° C for 24 hours while awaiting handling. The determination of the enzymatic activity of G6PD was carried out by the spectrophotometric method using a commercially available kit. 02/108 subjects studied (ie a prevalence of 1.85%) were deficient in our study. These two deficit were mostly young and between 18-25 years old only male. They were originally from the plateau and observed in the blood transfusion station (PTS) of the Talangai referral hospital. The mean hemoglobin value in normal subjects was 12.53 g / dl in men and 11.3 g / dl in women. In contrast, the average hemoglobin value was 10.45 g / dl in the deficient patients. The mean value of G6PD enzyme activity in normal subjects was 11.7 IU / g hemoglobin in men, 11.54 IU / g hemoglobin in women and in deficient subjects was 2.83 IU / g hemoglobin. As the results of our study show, G6PD deficiency affects hemoglobin levels.

Accurate diagnosis of glucose-6-phosphate dehydrogenase (G6PD) deficiency is required to avoid the risk of acute hemolysis associated with 8-aminoquinoline treatment. The performance of the BinaxNOW G6PD test compared with the quantitative spectrophotometric analysis of G6PD activity was assessed in 356 Plasmodium vivax-infected subjects in Brazil, Peru, Thailand, and India. In the quantitative assay, the median G6PD activity was 8.81 U/g hemoglobin (range = 0.05-20.19), with 11 (3%) subjects identified as deficient. Sensitivity of the BinaxNOW G6PD to detect deficient subjects was 54.5% (6 of 11), and specificity was 100% (345 of 345). Room temperatures inadvertently falling outside the range required to perform the rapid test (18-25 degrees C) together with subtlety of color change and insufficient training could partially explain the low sensitivity found. Ensuring safe use of 8-aminoquinolines depends on additional development of simple, highly sensitive G6PD deficiency diagnostic tests suitable for routine use in malaria-endemic areas.

Equine carbonic anhydrase isozymes (CA-I and CA-II) were purified from erythrocytes by several column chromatography. Polyclonal anti-CA-I and anti-CA-II sera were produced in rabbits. Sensitive competitive enzyme-linked immunosorbent assays (ELISA) were established to determine the developmental changes in CA-I and CA-II levels in equine erythrocytes. Concentrations of CA-I and CA-II in erythrocytes from 150 clinically normal thoroughbreds (123 racehorses and 27 riding horses) were determined by ELISA. Mean (+/- SD) concentrations of CA-I and CA-II in racehorses were 1.70 +/- 0.48 and 0.94 +/- 0.13 mg/g hemoglobin (Hb), respectively. Mean concentrations of CA-I and CA-II in riding horses were 2.34 +/- 0.52 and 0.76 +/- 0.08 mg/g Hb, respectively. When the CA levels in racehorses and riding horses were compared, the CA-I level in riding horses was higher than that in racehorses (p=0.01). The CA-II level in racehorses was higher than that in riding horses (p=0.02). These data suggest that the levels of CA isozymes in erythrocytes of racehorses were influenced by chronic physical stress. The CA-I concentration in erythrocytes of 2-month-old horses was approximately 0.25 mg/g Hb. The CA-I level noticeably increased during the first year of life and approached normal adult levels by 2 years. The CA-II level decreased slightly with age, indicating different regulation of CA-I and CA-II expression during development.

An additive solution for the storage of red blood cells was evaluated for use in dogs. Blood collected from 6 dogs was processed into packed red blood cells and stored for 35 days in the additive solution Nutricel (Miles, Inc, Pharmaceutical Division, West Haven, CT). Packed red blood cells stored in citrate-phosphate-dextrose-adenine (CPDA-1; Fenwal Laboratories, Baxter Health Care Corp, Deerfield, IL) also were evaluated for comparison. Red blood cell 2,3-diphosphoglycerate (2,3-DPG) concentration, adenosine triphosphate (ATP) concentration, percentage hemolysis, and pH were determined. The red blood cell post-transfusion viability (PTV) after 35 days of storage was assessed with both single-labeled chromium 51 (51Cr) and double-labeled technetium 99m/chromium 51 (99mTc/51Cr) techniques. Mean ATP concentration and percentage hemolysis of the cells stored in Nutricel were 1.1 mumol/g hemoglobin (Hb) and 0.28% respectively and did not differ significantly (P < .05) from the values of 1.0 mumol/g Hb and 0.33% from the CPDA.1-stored red blood cells. The mean pH of red blood cells stored in Nutricel was 6.19, which was significantly lower than the pH of 6.47 for cells stored in CPDA-1. The mean 2,3-DPG concentration of red blood cells stored in Nutricel was significantly higher at 10.1 mumol/g Hb than the 2,3-DPG concentration of 3.4 mumol/g Hb for cells stored in CPDA-1. The mean PTV of canine red blood cells stored in Nutricel for 35 days was 85% with 51Cr and 90% with 99mTc/51Cr. This was significantly higher than the mean PTVs of 38% and 36% for the CPDA-1 stored cells as assessed with 51Cr and 99mTc/51Cr techniques, respectively. It was concluded that 35-day-old canine red blood cells stored in Nutricel are of acceptable quality for transfusion purposes.

Aim: Our purpose was to determine the frequency of glucose-6-phosphate dehydrogenase (G6PD) deficiency in blood donors of Jahrom, Iran. Methods: Blood samples were obtained from the 706 blood donors who referred to Jahrom Blood Transfusion Organization between June and September 2008. Enzyme assays were performed three times, on day 1 (day of blood sampling), 7 and 45 days after blood storage. G6PD activity was measured using a quantitative assay. Results: A total of 706 samples were examined. 97.7% males and 2.3% females, mean age 32.6 years (18–64 years). Based on enzyme activity less than 1.62 IU/g hemoglobin (Hb), prevalence of G6PD deficiency in three separate measurements was 16.3, 19.1 and 33.3% respectively. Four percent of donor had an enzyme level of zero on day 1 which increased to 7.4 and 10.7% on the seventh and the forty-fifth days. Conclusion: The results of this study support the screening for G6PD as part of the routine workup of blood donors in areas with a high prevalence of G6PD deficiency.

Abstract 3437 Each unit of blood donated is processed and stored individually resulting in wide variability in the amount of red blood cells (RBCs) collected, RBC properties and in the 24 hour post-transfusion RBC survivability. As a result, each unit differs in its RBC content, its ability to deliver oxygen and potentially its effects on the recipient. The goal of this study was to investigate the effect of pooling and the use of novel additive solutions in producing a more uniform, standardized RBC product. RBCs were either stored in an approved additive solution with a 42 day shelf life (AS-3) or a solution which potentially enables an extended 50 to 100 day shelf life (EAS-81). The storage characteristics of individual RBC units were compared to pooled RBC products prepared by the same units and suspended in AS-3 and EAS-81 with comparable final hematocrits. Two sets of 5 individual leukoreduced group A, D-positive blood group RBC units were divided and stored as individual RBC products or as pooled products. Individual and pooled units tested as group A, D-positive and had a negative antibody screen. Quality indices were measured (ATP, 2,3 DPG and hemolysis) for all individual and pooled units weekly for 5 weeks. After 5 weeks of storage RBC units stored in EAS-81 showed higher ATP concentrations (2.53–4.20 μmole/g hemoglobin) compared to units stored in AS-3 solution (1.53–1.98 μmole/g hemoglobin). ATP concentration values of pooled units of EAS81:AS-3 (3.35:1.77 μmole/g hemoglobin) were close to average (3.24:1.64 μmole/g hemoglobin) values obtained from 5 individual unit concentrations (Figure). Next, after 5 weeks hemolysis was higher in EAS-81 (0.57–0.94% hemolysis) as compared to AS-3 solution (0.18–0.35% hemolysis); possibly due to cells being passed through an additional leukoreduction filter. The hemolysis values of pooled units of EAS-81: AS-3 (0.51:0.27%) was lower for EAS-81 but the same for AS-3 as the average values obtained from individual units (0.80:0.27%). Lastly, after 2 weeks of storage 2,3 DPG values obtained were lower for AS-3 units (2.71–3.84 μmole/g hemoglobin) as compared to EAS-81 units (3.40–7.10 μmole/g hemoglobin). 2,3-DPG concentration of pooled unit EAS-81:AS-3 (7.10:3.38 μmole/g hemoglobin) were higher compared to average of individual unit (4.74:3.19 μmole/g hemoglobin). 2,3 DPG concentration was negligible for 3 week measurement for all the units. In conclusion, RBCs stored in EAS-81 have higher ATP and 2,3-DPG levels than units stored in AS-3; pooled units maintain ABO/D typing without new antibody or antigen formation, and appear to have slightly better storage characteristics than individually stored units. Thus, our data support pooling RBC units is a valid method to provide more uniform RBC products with better storage characteristics, especially if stored in EAS-81, in the future. Disclosures: Stassinopoulos: Cerus: Employment, Equity Ownership, Patents & Royalties. Hess:U. S. Army, Hemerus Medical, LLC: Membership on an entity's Board of Directors or advisory committees, Patents & Royalties.

The relationship between the blood NAD and NADP levels and the niacin equivalent intake was investigated using women students. The daily niacin equivalent intake was 151±64μmol (18.5±7.5mg in terms of nicotinamide, mean±SD). The blood NAD and NADP levels were 34.7±7.0 and 19.3±2.3nmol/ml whole blood, respectively. The blood NAD level was proportional to the blood NADP level. Both blood levels were also proportional to the hemoglobin content, and the NAD and NADP levels per g hemoglobin were 233.7±44.7 and 156.3±25.5nmol. The blood NAD and NADP levels increased with an increase in the niacin equivalent intake. From these results, it is concluded that the blood total pyridine nucleotide level (NAD+NADP) reflects the niacin equivalent intake. Accordingly, the blood pyridine nucleotide level may be useful as an index for niacin nutrition in humans.

A sensitive and reproducible method for quantitation of polyamines in erythrocytes by high-performance liquid chromatography (HPLC) is described. Spermidine and spermine are first converted to fluorescent dansyl derivatives and are then separated in less than 15 minutes on a C18 reverse-phase cartridge using methanol in water mobile phase. Sensitivity of the method is 20 pmol with recoveries that average 96% and 95% for spermidine and spermine, respectively. Precision study revealed an average mean coefficient of variation of 5.42 ± 2.95 (mean ± S.D.) from four levels of the polyamines (8.2 to 354.9 nmol/g hemoglobin [Hgb]). The normal ranges for 30 apparently healthy men and women, aged 20–60 years, were 2.9–33.7 nmol/g Hgb (± 2 S.D. from mean) for spermidine and 0-20.9 nmol/g Hgb for spermine. The quantity of putrescine was negligible. Feasibility of this method was evaluated in serial specimens from an advanced-stage patient with prostate cancer who was receiving multiple-modality therapy. Results revealed that this HPLC method can be used in quantifying circulating polyamines in clinical specimens.

Extract: We have found a genetic trait of arginase deficiency in erythrocytes (RBC) of Macaca fascicularis (crabeater macaque). Measurements of arginase activities and arginine concentrations in RBC, and information derived from breeding experiments indicated that arginase activities in RBC are 12.59 mmoles/g hemoglobin (Hb)/hr (range: 8.36–15.24) in normal monkeys, 4.40 mmoles/g Hb/hr (sd ± 2.37) in hetero-zygotes for this trait, and less than 0.05 mmoles/g Hb/hr in homozygotes for this trait. Concentrations of arginine in erythrocytes are less than 0.01 $mUmoles/ml packed cells in both normal and heterozygous monkeys, and 0.975 $mUmoles/ml packed cells (sd ± 0.556) in homozygous monkeys. There are no significant differences in plasma amino acids and hepatic arginase activity in any animal. Speculation: Study of arginase activity in RBC will be expanded to include a large number of monkeys in order to determine the incidence of arginase deficiency in RBC in this species. The possibility of using these monkeys as an animal model for study of human disease arginase deficiency (e.g., genetic engineering) will be explored.

A modification is described of the automated determination of 2,3-diphosphoglycerate (DPG) in blood [Grisolia, S., et al., Anal. Biochem. 31, 235 (1969)]. Modifications in the manifold result in a sensitive, noise-free, rapid system and the modifications in the preparations of the reagents ensure stability of the diluted standards and blood samples for at least three weeks. Samples are run at the rate of 60/h and sample size can be as small as 5 µl of whole blood. The coefficient of variation of the overall determination of automated DPG and manual hemoglobin is 3.6% and the SD is ±0.77 µmol/g Hb. The normal range is 14.6 ± 2.2 (SD) µmol/g hemoglobin.

Plasma selenium concentration and glutathione peroxidase activity in red blood cells were determined in subjects from different age groups. The selenium level (mean +/- SD) found in infancy (0 to 6 months) was 2 +/- 0.6 micrograms/dl, with the lowest value of 1 microgram/dl observed in a 4-month-old infant. These levels were significantly lower (p less than 0.001) than the value of 9.5 +/- 1.1 micrograms/dl found in the adult group and 7.7 +/- 1.3 micrograms/dl found in the group of older children (2 to 15 yr). Younger children (6 to 24 months of age) had intermediate levels of 5 +/- 1.2 micrograms/dl. When the data were plotted on a logarithmic scale as a function of age, the figure shows clearly that the plasma selenium levels increase steadily with age throughout life after an initial drop at 60 to 90 days. There was a satisfactory correlation between the plasma selenium concentration and the enzyme glutathione peroxidase activity in the red blood cells (Spearman's p = 0.45, p less than 0.005). Although very low selenium values were observed, the enzyme glutathione peroxidase activity remained above 10 U/g hemoglobin (with only one exception) in all patients.

OBJECTIVES. We conducted a clinic-based study of erythrocyte cholinesterase levels, pesticide exposures, and health effects among farmworkers and nonfarmworkers to determine risks for exposure and associated morbidity. METHODS. Two hundred two farmworkers and 42 nonfarmworkers were recruited sequentially at two community health centers. Erythrocyte cholinesterase levels were measured colorimetrically. Questionnaires obtained data on demographics, occupational history, exposures, and symptoms. RESULTS. Cholinesterase levels were significantly lower among farmworkers (30.28 U/g hemoglobin) than among nonfarmworkers (32.3 U/g hemoglobin). Twelve percent of farmworkers, but no nonfarmworkers, had very low levels. Farmworkers applying pesticides also had lower cholinesterase levels. One half of farmworkers reported being sprayed by pesticides and working in fields with an obvious chemical smell. Of reported symptoms, only diarrhea was associated with cholinesterase levels. Reported exposures, however, were strongly associated with symptoms. CONCLUSIONS. Farmworkers reported many pesticide exposures that violate state and federal regulations. Farmworkers had cholinesterase levels significantly lower than those of nonfarmworkers, although only spraying pesticides was associated with very low levels.

The deficiency was first found by Ogasawara et al /BBRC 1984, 122, 1344 - 1349, Hum.Genet. 1987, 75, 15 - 18/ to occur in Japan, Seoul and Taipei as one heterozygote in about 30 of the population. In order to find the frequency of the defect in the european area, 410 blood samples have been collected from the blood donors in the county blood center in Gdansk /Poland/. The mean AMP deaminase activity in the erythrocytes was found to be 8.3 μmole ammonia/min per g hemoglobin, which is about 30% less than found in Japan but exactly the same which was found in U.S.A. by Campbell et al. /Clin.Chim Acta 1977, 79, 379 - 385/. Among the 410 individuals investigated 26 displayed half or less of the mean erythrocyte AMP deaminase activity. This could mean that the frequency of the mutant gene is one heterozygote in about 16 of the population in Poland. One case of a complete deficiency of the erythrocyte AMP deaminase activity was found in the family member of one of the heterozygotes. All the deficient subjects appeared to be perfectly healthy, there were no evidence of either anaemia or hemolysis.

Peroxide-induced erythrocyte malonyldialdehyde formation (RBC-MDA) has been used as a measure of polyunsaturated fat autooxidation, a process believed to alter membrane transport and enzyme function. Evidence for this process was sought in erythrocytes of RS patients using the method of Bidder (Life Sciences 30:1021, 1980). For 7 children with stage II-IV RS, RBC-MDA was 432 ± 36 nmoles/g hemoglobin on admission, 323 ± 25 at the nadir, and 478 ± 67 1-4 months after RS. Nadir values differed significantly from admission and from values for 7 siblings(431 ± 28), 5 pediatric intensive care unit (ICU) patients(517 ± 32)and 5 adults (530 ± 40). Corresponding values for plasma vitamin E in RS patients were 556 ± 37 mcg/dl at admission, 473±36 at the nadir, and 905±30 after RS. Nadir values were significantly different from recovery, siblings 1001±43, and ICU patients 740±98. Conclusion: The observed decrease in RBC-MDA in RS could be explained by decreased substrate (if RBC polyunsaturated fatty acids had undergone peroxidative degeneration) or, less likely, given the low plasma vitamin E, increased antioxidant activity.