Interstrain differences in red cell enzyme activities in mice and rats.

1. Interstrain differences in red blood cell enzyme activities were studied in mice (BALB/c, C57BL/6, C3H/He, DBA/2 and ddY) and rats (Donryu, F344/N, SD, Wistar and Wistar/ST), and were also compared with hamster, guinea-pig and rabbit. 2. The enzyme activities measured were: glutathione S-transferase (GST), glucose-6-phosphate dehydrogenase (G-6-PD), 6-phosphogluconate dehydrogenase (6-PGD), NADPH-diaphorase (ND), hexokinase (Hx), glutamate oxaloacetate transaminase (GOT), lactate dehydrogenase (LDH) and acetylcholinesterase (AChE). 3. There were marked variations in the activities of some red cell enzymes (e.g. GST, Hx, ND), while others (e.g. G-6-PD, 6-PGD) were much less variable both within different strains and species.     

Blood genetic markers in the Chinese of two eastern provinces

A total of 205 Han Chinese from two eastern provinces (155 from Fujien and 50 from Hopeh) were tested for the distribution of six blood groups--A1A2BO, MN, Rhesus (CcDEe), Lewisa, Kell (Kk) and Fya--four serum proteins--albumin and haptoglobin types; transferrin and group-specific component subtypes--haemoglobin, and twelve red cell enzyme systems--glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, lactate and malate dehydrogenases; acid phosphatase, esterate-D, glyoxalase I, adenylate kinase, glucose-phosphate isomerase, phosphoglucomutase (locus 2), and superoxide dismutase types; and phosphoglucomutase (locus 1) subtypes. The frequencies of blood groups were more or less within the reported frequencies in the Chinese. However the frequency of le was much lower in the present series. The Chinese are characterized by low p1, Ro, k, le, and a high Fya in general. P2 was lacking in the Chinese. There were some differences in the blood group frequencies in the two provinces. The frequencies of Hp alleles; Tf and Gc subtypes show characteristic mongoloid features with high Hp1, TfD, and GcIF. The frequency of TFC2 was higher in the Fujien province than that in Hopeh. At the hemoglobin locus only one Hb AD was detected, while the frequency of the beta-thalassemia trait was 0.03. No red cell G6PD deficiency or variant was detected. The distribution of red cell enzymes showed Mongoloid characteristics with low PGDC, AK2, ESD1, GLO1, and higher pa. PGM1 subtypes also had Mongoloid characteristics with lower PGM2+ and higher PGM2-. The phenotypic distribution of all the fifteen polymorphic loci was at Hardy-Weinberg equilibrium in both the Chinese populations.     

Effect of polychlorinated biphenyls (Delor 103) on haematological and enzyme parameters of the rainbow trout Oncorhynchus mykiss

Rainbow trout at a weight of 223+/-12 g (mean+/-SD) were experimentally injected with a technical mixture of Delor 103 to evaluate the red blood cell indices (red blood cell count, haematocrit, haemoglobin, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration) and some biochemical and enzyme parameters of the blood plasma (total protein, glucose, inorganic phosphate, total calcium, sodium, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase). Delor 103, administered by the i.p. route at a concentration of 0.24 g kg(-1) 120 h(-1), caused an increase in the red blood cell counts, haematocrit values, haemoglobin concentrations, inorganic phosphate, alanine aminotransferase and lactate dehydrogenase. The sodium level fell. The fish injected with Delor 103 showed a relative decrease in the lymphocyte count and a relative increase in the count of neutrophile band forms.     

Association of red cell glucose-6-phosphate dehydrogenase with haemoglobinopathies

A total of 1,112 randomly selected Saudi Arabs, of both sexes, living in Jeddah and the surrounding areas were screened for the phenotypic distribution of red cell glucose-6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD). They were also investigated for haemoglobin and for thalassaemia. Phenotyping of the haemoglobins and the red cell enzymes was carried out by starch gel electrophoresis and the dye-decolouration screening test, while the investigation for thalassaemia was carried out by globin-chain biosynthesis, followed by column chromatography. The red cell Gd- alleles were significantly associated with the sickle-cell gene in both the males (chi 2(1): AS-28.80; SS-4.89) and females (chi 2(1): AS-10.99; SS-13.16). A similar association was also observed between G6PD deficiency and thalassaemias in males (chi 2(1): alpha-thalassaemia - 3.13; beta-thalassaemia - 11.06) and females (chi 2(1): alpha-thalassaemia - 6.63). However, no such association was detected between red cell 6PGD types and haemoglobin genes. The results suggest that the red cell G6PD deficiency, sickle-cell and thalassaemia genes might have evolved as a result of the same ecological factor, probably malaria.     

The relationship between red cell aging and enzyme activities in experimental animals.

1. The relationship between red cell aging and enzyme activities was studied in rabbit, guinea-pig, hamster, rats (F344/N and SD), and mice (BALB/c and DBA/2). 2. The activities of six enzymes: glucose-6-phosphate dehydrogenase (G-6-PD), 6-phosphogluconate dehydrogenase (6-PGD), hexokinase (Hx), glutamate oxaloacetate transminase (GOT), lactate dehydrogenase (LDH) and acetylcholinesterase (AChE), were measured in the red cells of different ages which were obtained either by centrifugation or experimental anaemia. 3. Hx, AChE and GOT activities were much higher in younger red cells than in older cells, hence the activities of these enzymes may be used as an indicator of age of the cells.     

Localization of human gene loci using spontaneous chromosome rearrangements in human-Chinese hamster somatic cell hybrids.

Analysis of human-Chinese hamster somatic cell hybrids with spontaneously derived chromosome structural changes has provided data for the regional and subregional localization of gene loci which have previously been assigned to human chromosomes 2, 12, and X. Correlation of the expression of human gene loci with the human chromosome complements present in somatic cell hybrids indicates that the cytoplasmic malate dehydrogenase (MDH1) locus is in the 2p23yields2pter region, and red cell acid phosphatase (AcP1) is at or adjacent to 2p23. The cytoplasmic isocitrate dehydrogenase (IDH1) locus is at or adjacent to 2q11, peptidase B (Pep B) is at or adjacent to 12q21, lactate dehydrogenase B (LDH B) is in the 12q21yiedls12pter region, glucose-6-phosphate dehydrogenase (G6PD) is in the Xq24yieldsXqter region, and the gene loci for phosphoglycerate kinase (PGK), alpha-galactosidase (alpha-gal), and hypoxanthine guanine phosphoribosyltransferase (GPRT) are in the Xp21yieldsXq24 region.     

Red cell sequestration during high dose intravenous immunoglobulin in idiopathic thrombocytopenic purpura

The cellular interactions involved in the platelet response after immunoglobulin infusion in acute and chronic idiopathic thrombocytopenic purpura (ITP) are unknown. There have been a number of theories including the competitive inhibition of platelet-binding to macrophages by the preferential sequestration of immunoglobulin coated red cells. We report a study to examine this hypothesis. Adult acute and chronic patients were given infusions of immunoglobulin at a rate of 0.4 g/kg body weight, daily for 5 days. Serum haptoglobin, lactate dehydrogenase and the absolute reticulocyte counts were monitored and no significant change in any value was seen during the period of study. A red cell survival was performed on four of the patients and no increase in the rate of red cell clearance occurred during the infusion period. We conclude from this that in these patients no significant degree of haemolysis was provoked by the infusion although this does not preclude this as a mechanism of action in some individuals.     

Study of the bovine erythrocyte. Enzymatic activities of the cell and its membrane

In bovine red cells, haemolysed and extensively washed, ten different enzyme activities were found to be present. The cells easily release glucose 6-phosphate dehydrogenase, glucose phosphate isomerase, fructose bisphosphate aldolase, and aspartate aminotransferase into the haemolysis medium. An important part of the last two enzymes and all the isocitrate dehydrogenase (NADP linked) are retained in the membrane. The levels of these enzymes in the membrane are strongly dependent on the age of the preparation. The optimal assay conditions have been defined for some of these enzymes. These findings are discussed in relation to red cell and membrane structure.     

Stress response and cytoskeletal proteins involved in erythrocyte membrane remodeling upon Plasmodium falciparum invasion are differentially carbonylated in G6PD A- deficiency

Multiple glucose-6-phosphate dehydrogenase (G6PD)-deficient alleles have reached polymorphic frequencies because of the protection they confer against malaria infection. A protection mechanism based on enhanced phagocytosis of parasitized G6PD-deficient erythrocytes that are oxidatively damaged is well accepted. Although an association of this phenotype with the impairment of the antioxidant defense in G6PD deficiency has been demonstrated, the dysfunctional pathway leading to membrane damage and modified exposure of the malaria-infected red cell to the host is not known. Thus, in this study, erythrocytes from the common African variant G6PD A- were used to analyze by redox proteomics the major oxidative changes occurring in the host membrane proteins during the intraerythrocytic development of Plasmodium falciparum, the most lethal malaria parasite. Fifteen carbonylated membrane proteins exclusively identified in infected G6PD A- red blood cells revealed selective oxidation of host proteins upon malarial infection. As a result, three pathways in the host erythrocyte were oxidatively damaged in G6PD A-: (1) traffic/assembly of exported parasite proteins in red cell cytoskeleton and surface, (2) oxidative stress defense proteins, and (3) stress response proteins. Additional identification of hemichromes associated with membrane proteins also supports a role for specific oxidative modifications in protection against malaria by G6PD polymorphisms.     

Haematology of the Australian eastern quoll, Dasyurus viverrinus--II. Red cell enzymes and metabolic intermediates

1. The activity of 21 red cell enzymes and three red cell metabolic intermediates were measured in adult Dasyurus viverrinus and compared with published data on other marsupials. 2. Phosphofructokinase (PFK), glyceraldehyde dehydrogenase (GAPD) and phosphoglycerate kinase (PGK) were elevated in comparison to other marsupials. 3. Enolase (ENO) and 2,3-diphosphoglycerate (2,3 DPG) were lower than in other marsupials.     

Genetic polymorphisms in the Kuwaiti Arabs

Blood samples were collected from 162 Kuwaiti Arabs. These samples were typed for the ABO, MNSs, Rh, Kell and Duffy blood group systems, serum protein haptoglobins, the red cell isoenzymes acid phosphatase, phosphoglucomutase (locus 1), adenylate kinase, 6-phosphogluconate dehydrogenase, and the lactate and malate dehydrogenase variants. Comparisons were made with serological findings for other Arab populations in the Arabian peninsula.     

四川南江两种水青冈种群遗传多样性初步研究

利用凝胶电泳法研究米心水青冈（Ｆａｇｕｓｅｎｇｌｅｒｉａｎａ）和巴山水青冈（Ｆ．ｐａｓｈａｎｉｃａ）２个种４个种群的遗传多样性。所测定的酶系统包括：过氧化物酶（ＰＸ１和ＰＸ２）,磷酸葡萄糖脱氢酶（ＰＧＤ）,酸性磷酸化酶（ＡＣＰ）,超氧物歧化酶（ＳＯＤ）,谷氨酸草酰乙酸转氨酶（ＧＯＴ１和ＧＯＴ２）,异柠檬酸脱氢酶（ＩＤＨ）,磷酸果糖异构酶（ＰＧＩ）,甲基萘醌还原酶（ＭＮＲ）,葡萄糖磷酸变位酶（ＰＧＭ１和ＰＧＭ２）和苹果酸脱氢酶（ＭＤＨ２）１０种酶系统。测定和分析了水青冈等位基因频率、遗传多样性、固定指数、Ｈａｒｄｙ－Ｗｅｉｎｂｅｒｇ平衡和遗传距离指标,为进一步研究水青冈属各种间的亲缘关系和进化提供了科学依据     

Multiple forms of formaldehyde dehydrogenase from human red blood cells

Red cell hemolysates from nonrelated Finns were analyzed by electrofocusing on polyacrylamide gel, and formaldehyde dehydrogenase (EC 1.2.1.1) was located by an activity-staining method. Three forms of the enzyme were constantly found for all the individuals studied but no variants were observed in this population (n = 217). Human liver also had three formaldehyde dehydrogenase forms with locations identical to those of the red cell formaldehyde dehydrogenase. Population genetic studies of formaldehyde dehydrogenase can easily be performed with red cell hemolysates with the techniques described here, and there is no need to use liver biopsy samples.     

十一个少数民族红细胞酸性磷酸酶、酯酶D、6-磷酸葡萄糖酸脱氢酶及谷丙转氨酶的遗传多态性

用淀粉凝胶电泳法对我国十一个少数民族红细胞酸性磷酸酶(AcP_1)、酯酶D(EsD)、6-磷酸葡萄糖酸脱氢酶(6-PGD)及谷丙转氨酶(GPT)的遗传多态性进行了研究,共调查了2272人。研究结果表明:侗、回、白、土家、苗、彝、藏、满、瑶、哈尼和布依等民族AcP_1~B基因频率依次为0.7835、0.7958、0.8137、0.7750、0.7624、0.8038、0.8075、0.8035、0.7725、0.6488和0.6896;EsD~1基因频率依次为0.6418、0.7315、0.6005、0.6025、0.6411、0.6411、0.6558、0.6305、0.6020、0.6023和0.6368;6-PGD~A基因频率依次为0.9279、0.9381、0.9387、0.9150、0.9356、0.9014、0.7764.0.8818、0.9851.0.9233和0.9410;GPT~1基因频率依次为0.4075、0.5367、0.5049、0.4824、0.5322、0.6106、0.6313、0.6400、0.3985、0.4930和0.3976。并对发现的变异型进行了讨论。     

Red cell lactic acid dehydrogenase in the family Macropodidae

Lactic acid dehydrogenase isoenzyme patterns from red cells of members of the family Macropodidae are described as they appear in disc acrylamide gels. The reproducibility of patterns, which are determined by the relative intensities of LDH isoenzymes, is noted. Of the 473 animals tested, 471 had red cell LDH patterns belonging to one of five patterns. The distribution of these five patterns among the different genera and species is discussed, and it is suggested that investigation of LDH patterns by this method might assist in the classification of members of the family Macropodidae.     

Glyceraldehyde-3-phosphate dehydrogenase of rat erythrocytes has no membrane component

In the course of studying mammalian erythrocytes we noted prominent differences in the red cells of the rat. Analysis of ghosts by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis showed that membranes of rat red cells were devoid of band 6 or the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (D-glyceraldehyde-3-phosphate: NAD+ oxidoreductase (phosphorylating), EC 1.2.1.12). Direct measurements of this enzyme showed that glyceraldehyde-3-phosphate dehydrogenase activity in rat erythrocytes was about 25% of that in human cells; all of the glyceraldehyde-3-phosphate dehydrogenase activity in rat erythrocytes was within the cytoplasm and none was membrane bound; and in the human red cell, about 1/3 of the enzyme activity was within the cytoplasm and 2/3 membrane bound. The release of glyceraldehyde-3-phosphate dehydrogenase from fresh rat erythrocytes immediately following saponin lysis was also determined using the rapid filtration technique recently described. The extrapolated zero-time intercepts of these reactions confirmed that, in the rat erythrocyte, none of the cellular glyceraldehyde-3-phosphate dehydrogenase was membrane bound. Failure of rat glyceraldehyde-3-phosphate dehydrogenase to bind to the membranes of the intact rat erythrocyte seems to be due to cytoplasmic metabolites which interact with the enzyme and render it incapable of binding to the membrane.     

Contribution of hydrogen peroxide to the cytotoxicity of green tea and red wines

Green tea and red wine are claimed to have health benefits because of their high content of polyphenolic compounds, but they have also been reported as mutagenic in some test systems. In this paper, we show that a commonly used cell culture medium, Dulbecco's modified Eagle's medium (DMEM), catalyses oxidation of green tea and red wines to generate H(2)O(2). The level of H(2)O(2) produced from green tea accounted for all of the cytotoxic effects of this beverage on PCl2 cells. By contrast, H(2)O(2) was only responsible for part of the cytotoxicity of the red wines examined. Our data illustrate the danger of extrapolating from cell culture studies to predict the effects of complex beverages in vivo.     

Characterization of a new continuous cell line from the flood water mosquito,Aedes vexans

A new cell line, UM-AVE1, was established from embryos of the mosquito Aedes vexans. Banding patterns for the isozymes lactate dehydrogenase (LDH), malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH), xanthine dehydrogenase (XDH), and esterases were compared with those of larval Aedes vexans tissues as well as those of four other mosquito cell lines and one moth cell line. Karyotype analyses confirmed that the dipteran cell lines were not contaminated with lepidopteran cells, because in all mosquito lines the modal number of chromosomes was 6 (=2n) or 7. Isozyme electrophoresis established a specific profile for each cell line. Two isozymes present in UM-AVE1 (LDH, IDH) were not detected in larvae; this could be a reflection of the different stages used for cell line isolation and enzyme analysis, or lability of sample preparations. It is significant that extracts from UM-AVE1 cells and Aedes vexans larvae had an identical double band for XDH, while all other cell lines examined exhibited only a single band.     

Genetic regulation of glucose 6-phosphate dehydrogenase activity in the inbred mouse

The erythrocyte glucose 6-phosphate dehydrogenase activity characteristic of each of 16 inbred mouse strains falls into one of three distinct classes. Strains C57L/J and C57BR/cdJ represent the low activity class: strains A/J and A/HeJ represent the high activity class; other strains have intermediate activities. There is no evidence that structural variation is responsible for the variation in G6PD activity, since partially purified enzyme from each class has the same thermal stability, pH-activity profile, Michaelis constants for G6P and NADP, electrophoretic mobility, and activity using 2-deoxy d-glucose 6-phosphate as substrate. The activities of 6-phosphogluconate dehydrogenase and glucose phosphate isomerase do not differ in erythrocytes of the three G6PD activity classes. Young red cells have higher G6PD activities than old red cells and there is evidence that the intracellular stability of the enzyme is reduced in red cells of strain C57L/J. G6PD activities in kidney and skeletal and cardiac muscle from animals with low red cell G6PD are slightly lower than the activities in kidney and muscle from animals with high red cell G6PD activity. The quantitative differences in red cell G6PD activity are not regulated by X-linked genes, but by alleles at two or more autosomal loci. A simple genetic model is proposed in which alleles at two unlinked, autosomal loci, called Gdr-1 and Gdr-2 regulate G6PD activity in the mouse erythrocyte.