The effect of cardiac disease on hemoglobin-oxygen binding.

The relation between degree of cardiac functional impairment and changes in hemoglobin-oxygen affinity and 2,3-diphosphoglycerate (2,3-DPG) has been studied in 39 patients with noncyanotic heart disease. A progressive decline in hemoglobin-oxygen affinity was found with worsening cardiac function as assessed by cardiac index, arteriovenous oxygen (A-V O(2)) difference, and cardiac symptoms; this alteration in hemoglobin-oxygen binding represents a significant mechanism for adaptation to the limited oxygen supply imposed by the cardiac lesion. The highly significant correlation of mixed venous blood oxygen saturation (S[unk]V(VO2)) with 2,3-DPG and the position of the oxygen dissociation curve suggests that the level of deoxygenated hemoglobin is an important in vivo regulator of hemoglobin-oxygen affinity.

Changing patterns of dietary iron overload in black South Africans.

Over the last 17 years there has been a significant reduction in the prevalence and severity of dietary iron overload in urban blacks of Johannesburg. This is attributed to a decrease in the consumption of traditionally brewed beer of high iron content over this period. A 40% reduction was found in hepatic iron concentrations measured in necropsy specimens from 248 male patients who died in 1976 as compared with 220 who died in 1959 and 1960. While hepatic iron concentrations rose with age in both groups there was no evidence of iron accumulation during the period between the two studies. Hepatic iron concentrations measured in 345 female subjects were many fold less than those of males and the 1976 group did not differ significantly from the 1959 to 1960 group. A paradoxical increase in the prevalence of portal fibrosis and cirrhosis was seen and may be due to the effects of increased ingestion of spirits and fortified wine in recent years. Iron overload was significantly greater in males with carcinoma of the esophagus and in those with idiopathic heart failure when compared to subjects who died of other causes. This suggests excessive exposure of these subjects to traditionally brewed beverages and the adulterants present in them.

The relative effect of ascorbic acid on iron absorption from soy-based and milk-based infant formulas.

The effect of varying concentrations of ascorbic acid on the absorption of iron from a soy-based infant milk formula containing 6 mg iron/100 g was examined in 64 adult Indian females using the extrinsic radioactive tag method. The corrected geometric mean absorption from the basic soy formula was only 1.8%. Addition of ascorbic acid in a concentration of 40 mg/100 g, did not significantly increase absorption (3.3%; t = 1.8, p greater than 0.07) but raising the concentration to 80 mg/100 g did so (6.9%; t = 2.4, p less than 0.02). No further significant increase was noted when the concentration of ascorbic acid was increased to 160 mg/100 g (7.7%; t = 0.4, p greater than 0.7). The inhibitory effect of soy on iron absorption was further demonstrated by a direct comparison between the soy-based formula and a similar product based on cows' milk. The comparison was made at two concentrations of ascorbic acid. At 40 mg/100 g the geometric mean iron absorption from the soy formula was 2.4% compared with 5.3% from the milk formula (t = 2.8, p less than 0.02), while the corresponding values at 80 mg ascorbic acid/100 g were 7.2 and 19.5%, respectively (t = 3.4, p less than 0.02). The present results confirm the marked inhibitory effect of soy protein on iron absorption and calculations from the absorption figures suggest that such formulas should contain at least 12 mg/100 g iron together with ascorbic acid in a molar ratio of approximately 4:1 if they are to be adequate in terms of iron nutrition.

Storage iron in "muscle".

In Bantu subjects with iron overload iron is visible in skeletal muscle cells and in the tissue histiocytes which lie between these cells. In the present study the concentrations of ;muscle' iron were measured chemically in subjects with varying hepatic storage iron concentrations. The results indicate that the concentrations of storage iron in ;muscle' are much lower than those in the liver. However, the muscle mass is so large that the total amount of iron present is at least equal to that in the liver in subjects with normal body stores. The concentrations of iron in ;muscle' are raised in subjects with iron overload but the degree to which they rise is far less than occurs in the liver; a thirtyfold increase in hepatic iron concentrations is associated with only a sixfold increase in ;muscle' iron. Experiments in rats revealed that storage iron in ;muscle' represents a relatively non-miscible pool which responds very little to acute changes in the iron environment.

Changes in oxygen delivery during hemodialysis.

The effect of hemodialysis on oxygen delivery has been assessed in nine patients. During hemodialysis the arterial PO2 decreased from 76.89 mm Hg to 69.03 mm Hg. There was also an increase in pH from 7.39 to 7.49 which resulted in an increase in the affinity of the blood for oxygen. In the absence of compensatory changes this could result in a 17.9% decrease in oxygen delivery. This could be hazardous in patients who are already anemic particularly in the presence of cardiac or respiratory insufficiency.

Distribution of erythrocyte nucleotides in pyrimidine 5'-nucleotidase deficiency.

In pyrimidine 5'-nucleotidase deficiency, erythrocytes contain elevated levels of pyrimidine nucleotides. The composition of this nucleotide pool was examined by ion exchange chromatography on Dowex formate columns using a linear ammonium formate elution gradient. In contradistinction to normal erythrocytes, adenine nucleotides accounted for only 32% of the nucleotide pool. The remainder consisted of 50% cytidine and 16% uridine nucleotides. The remaining 2% was not identified. The most abundant compound appeared to be UDP glucose whilst high levels of CTP, CMP and an unidentified cytidine compound less polar than CMP accounted for most of the cytidine nucleotide pool. The possibility that the abnormal nucleotides were due to an elevated reticulocyte count was excluded and it was also shown that erythrocytes from subjects heterozygous for pyrimidine 5'-nucleotidase deficiency did not have detectable levels of the abnormal nucleotides.

Some properties of human erythrocyte pyrimidine 5'-nucleotidase.

In haemolysates human erythrocyte pyrimidine 5'-nucleotidase had a single optimum at pH 7.2 with CMP and 6.75 with UMP as substrate. The purified enzyme showed two pH optima at pH 6.25 and 7.2 with UMP as substrate. The enzyme was inhibited by both its products - inorganic phosphate and pyrimidine nucleoside. The inhibition by inorganic phosphate appeared to be non-competitive with Ki = 1.5 mM. Contrary to previous reports adenosine and inosine did not inhibit the enzyme.

Erythrocyte pyrimidine 5'-nucleotidase.

In this study 31 family members of a patient with erythrocyte pyrimidine 5'-nucleotidase deficiency were studied. The activity of this enzyme in their erythrocytes is compared with levels in normal subjects and the problems surrounding heterozygote detection are discussed. The mean erythrocyte pyrimidine 5'-nucleotidase activity in 158 normal fresh blood samples was 130 +/- SD 29.8 mU/gHb. There was no significant difference between males and females. The enzyme level in a patient with non-spherocytic haemolytic anaemia was 6 mU/gHb. Of 31 relatives of the enzyme deficient patient examined five were clearly heterozygous for the enzyme defect. Their enzyme levels were below 50 mU/gHb. The father, who is an obligatory heterozygote, had an enzyme level of 87 mU/gHb which falls within the low values of the normal range. The distribution of enzyme activity in 26 family members having enzyme activities greater than 50 mU/gHb suggests that six of these may be carriers of the defective gene. We were unable to identify carriers by enzyme kinetic studies, electrophoresis, chromatographic examination of acid extractable nucleotides or measurement of enzyme levels in young and old erythrocyte populations. The last-mentioned technique showed that erythrocyte 5'-nucleotidase activity in reticulocytes may be as high as 1785 mU/gHb and declines rapidly as the cell ages reaching about 50 mU/gHb in the oldest cells. Blood samples which had been stored frozen were examined to see whether such samples were satisfactory for population studies. The mean enzyme activity 151 blood samples stored frozen for more than 12 months was 153+/-SD 44.7 mU/gHb. The increase in enzyme levels in the frozen samples appears to be greatest in samples showing haemolysis. In spite of the increased enzyme level frozen samples could be used to detect subjects with enzyme deficiency and some heterozygotes.

Oxygen affinity of avian blood.

Oxygen dissociation curves plotted directly on whole blood during oxygenation using a Radiometer DCA 1 gave P50 values of 39.2 +/- 2.2 mm Hg and 51.3 +/- 6.2 mm Hg for pigeon and chicken blood, respectively, at 41 degrees C, pH 7.5 and PCO2 29 mm Hg. These values agree well with earlier results obtained by a variety of methods; however, they do conflict with recent findings of a much higher oxygen affinity in avian blood which were obtained using a different plotting method. It was shown that with the method used in this paper the high metabolic rate of avian blood has little effect on the plotted curve at saturations below 50%. At higher saturations O2 consumption of red cells affects a progressive shift of the curve to the left so that to obtain accurate curves at high saturation it is necessary to correct the plotted curves for the oxygen utilization.

Purification and properties of human erythrocyte pyrimidine 5'-nucleotidase.

A 250,000-fold purification of pyrimidine 5'-nucleotidase from human erythrocytes has been achieved using a combination of DEAE-cellulose chromatography, ammonium sulfate fractionation, gel filtration, and isoelectric focusing. Polyacrylamide disc and starch gel electrophoresis of the purified material show two strong protein bands. On starch gel these bands exhibited pyrimidine 5'-nucleotidase activity. Two faint protein bands devoid of enzyme activity were also found in the case of polyacrylamide electrophoresis. The enzyme has a pH optimum at 7.5 and is most stable between pH 6 and 7.5. The enzyme has pI of 5.0 and a molecular weight of 28,000 by gel filtration. The Km of the purified enzyme was 10 muM, compared to 40 muM when measured in hemolysate. The higher Km in the hemolysate is due to the presence of an inhibitor. Inorganic phosphate was shown to be a competitive inhibitor of pyrimidine 5'-nucleotidase and inorganic phosphate in the hemolysate may be responsible for increasing the Km of the enzyme for the substrate cytidine monophosphate.

Increased erythrocytic diphosphoglycerate in megaloblastic anaemia. A compensatory mechanism?

The levels of erythrocytic 2,3-diphosphoglyceric acid (2,3-DPG) in a group of patients with megaloblastic anaemia were compared with levels in an iron-deficient subject. The megaloblastic cells appeared to have 2 mumol more 2,3-DPG per g Hb than the iron-deficient cells, at the average haemoglobin level found in the megaloblastic group. The anticipated adaptive decrease in the oxygen affinity of the blood was not apparent when the relationship of P50 (7,4) to haemoglobin level was examined. This was shown to be due to a decreased mean corpuscular haemoglobin concentration (MCHC), which counteracted the effect of the increased 2,3-DPG.

Isolation of ferritin from human hepatocellular carcinoma.

Ferritin was extracted from human hepatocellular carcinoma tissue and purified using column chromatography, gradient gel electrophoresis and cadmium sulphate crystallization. DEAE cellulose chromatography showed a difference between hepatoma and normal liver ferritin, indicative of a more acidic isoferritin profile in the tumour. Column-purified and crystalline ferritin and that remaining in the mother-liquor after crystallization was subjected to isoelectric focusing. Hepatoma ferritin showed higher concentrations of acidic isoferritins than liver ferritin. This was most obvious with mother-liquor ferritin, as crystallization tended to select out more basic isoferritins. Subunit analysis of hepatoma and liver ferritin showed a higher proportion of heavy subunits in the tumour ferritin, in keeping with the presence of acidic isoferritins. An antibody against hepatoma mother-liquor ferritin was raised in rabbits. However, hepatoma ferritin proved to be antigenically identical with normal liver ferritin, and we were thus unable to develop a specific radioimmunoassay for hepatoma ferritin.

Erythrocyte zinc protoporphyrin determination for screening for lead exposure.

Free erythrocyte protoporphyrin (FEP), erythrocyte zinc protoporphyrin (EZP) and blood lead concentrations were measured in 150 male subjects, 110 of whom were employed in industries where lead exposure was possible. Eighty-seven per cent of these 110 workers had blood lead levels above the maximum safe level of 40 micrograms/dl, which suggests that the factory control of lead toxicity is inadequate. The AVIV dedicated front-face fluorometer (AVIV instruments, New Jersey, USA), which measures EZP, was evaluated as a simple, rapid method of screening for lead accumulation. EZP concentrations agreed well with conventional estimations of FEP (r = 0,987, P less than 0,0001), and both correlated well with the blood lead concentration (r = 0,558, P less than 0,0001 for EZP; r = 0,633, P less than 0,0001 for FEP). The specificity of the EZP level as a screening test was found to be high (98%) and the sensitivity moderately good (83%). In view of the ease and speed of operation of this portable fluorometer, it should prove useful as an aid to the detection of early lead intoxication.

Does plasma transferrin regulate iron absorption?

It has been suggested that transferrin that has recently donated its iron to receptor sites is 'activated' to take up iron more avidly from donor tissues. The hypothesis was tested in vitro in a system in which use was made of the different electrophoretic mobilities of normal and desialated transferrin. Recently desaturated transferrin and native apotransferrin were added in equal amounts to a solution of radioactive ferric citrate to produce various end saturations. The resultant mixture was electrophoresed on 5.4% polyacrilamide gel, which was then sliced and counted for 59Fe counts. The size of the 2 radioactive peaks was then compared and expressed as a ratio. Using this in vitro system no supporting evidence could be found for the hypothesis that diferric transferrin which has just donated its iron is able to bind available iron more avidly than native apotransferrin.

Haemolytic anaemia due to erythrocyte pyrimidine 5'-nucleotidase deficiency. Report of the first South African family.

The first South African case of haemolytic anaemia due to erythrocyte pyrimidine 5'-nucleotidase deficiency is reported. The anaemia is characterized by the presence of high erythrocyte pyrimidine nucleotide levels and marked basophilic stippling. The enzyme levels in 20 family members confirm an autosomal recessive mode of inheritance and illustrate the difficulty of diagnosing the carrier state.

Erythrocyte pyrimidine-5'-nucleotidase activity as a sensitive indicator of lead exposure.

Since the red cell enzyme pyrimidine-5'-nucleotidase (PN) is inhibited by lead, we examined the feasibility of using the activity of this enzyme as a measure of lead exposure. Erythrocyte PN activity was measured in 110 blood samples obtained from subjects working in industries which utilize lead and 40 control subjects. The measurements were then compared with a number of traditional indices of lead poisoning. These included blood and urine lead concentrations and free erythrocyte protoporphyrin, erythrocyte zinc protoporphyrin, urinary coproporphyrin and urinary delta-aminolaevulinic acid levels. There was a highly significant negative correlation between erythrocyte PN activity and blood lead concentration (-0,83; P less than 0,0001), which was greater than that for any of the other measurements. It was therefore concluded that erythrocyte PN activity is an excellent indicator of lead exposure.

A preliminary study of the treatment of trochanteric fractures of the femur with the Kenwright nail.

We present a preliminary study of the results of treatment of 106 fractures of the trochanteric region of the femur, using a new expanding compression nail, the Kenwright nail. The device obtains a secure hold in the femoral head and compresses the fracture line, to give very firm fixation, enabling patients to bear weight immediately with much more confidence than with conventional methods of fixation. Earlier discharge from hospital has been possible, with the release of valuable beds. In no case has the nail cut out. A similar study of its use in intracapsular fractures will be reported.