Alkali-resistant Hemoglobin Research Articles

Haematology of the australian eastern quoll, Dasyurus viverrinus

1. 1. A variety of haematological parameters were determined in adult Dasyurus viverrinus. 2. 2. Haemoglobin and red cell counts were high with a very low mean cell volume. 3. 3. Basophils are absent but the eosinophils contain small numbers of basophilic granules which may indicate a dual role for this cell. 4. 4. “Ring Form” leucocytes are present. 5. 5. Three types of red cell picture could be identified, some animals showing large numbers of spherocytes, spicule cells, and inclusion bodies. 6. 6. These cells resemble those found in some inherited human haemolytic anaemias but there was no evidence of haemolysis in the animals. 7. 7. An alkali resistant haemoglobin component is present.

595 BONE MARROW PROGENITOR CELLS IN ANEMIC AND NON-ANEMIC PATIENTS WITH FANCONI'S ANEMIA

Decreased bone marrow progenitor cells have been reported in patients with Fanconi's anemia. It is not known whether such a decrease occurs in forme fruste non-anemic Fanconi patients. Two patients with clinical Fanconi's anemia and their siblings with similar clinical features and chromosomal fragility but without anemia were studied. While one of the siblings had mild thrombo-cytopenia and slight increase in alkali resistant hemoglobin, the other showed no hematological abnormalities. The two patients with Fanconi's anemia and their non-anemic siblings, all showed marked decrease in myeloid and erythroid colonies: An unrelated non-Fanconi patient with increased chromosomal fragility showed normal myeloid and erythroid colonies (120/2× 105 cells and 51/105 cells respectively). The above findings suggest that in Fanconi's anemia, a deficiency of bone marrow progenitor cells is present despite normal hemogram and this test can be used for early detection of non-anemic patients.

Sickle cell syndromes: II. The sickle cell anemia-α-thalassemia syndrome

Five American black patients, ages 1 to 16 years, with the sickle cell anemia-alpha-thalassemia syndrome are described. Each patient had persistent microcytosis not explained by iron deficiency, and in each family the presence of alpha-thalassemia in combination with sickle cell trait was demonstrated in one of the parents. In one patient, in whom the diagnosis of sickle cell anemia was established at birth, an elevated level of Barts (gamma4) hemoglobin was also found. In these patients levels of alkali-resistant hemoglobin and reticulocyte counts were similar to those of sickle cell anemia patients of comparable age; however, stained smears of their peripheral blood rarely showed the presence of irreversibly sickled cells. No major ameliorative effect of the alpha-thalassemia on the clinical expression of the sickle cell disease of these patients was evident.

A new alkali-resistant hemoglobin alpha2J Oxford gammaF2 in a Sicilian baby girl with homozygous beta0 thalassemia

A 10-mo-old baby girl with homozygous beta0 thalassemia and alphaJOxford, presenting the clinical picture of homozygous beta thalassemia is described. Hemoglobin electrophoresis showed three bands: the first two with the mobilities of hemoglobin Hb A2 (1%) and Hb F (69%), respectively, the third migrating a little faster than Hb A (30%). About 30% of her alpha chains were J Oxford which, bound to her gamma chains, produced a new alkali-resistant hemoglobin, alpha2 J Oxford gamma F2, which has not been described previously. Hemoglobin synthesis in vitro showed the absence of beta chain synthesis and an alpha/non-alpha ratio of 2. The patient's father was heterozygous for both the Hb J Oxford and beta0 thalassemia genes, the mother a carrier of beta0 thalassemia; four other relatives were carriers of Hb J Oxford, and one was a carrier of beta thalassemia.

Studies on Camel Hemoglobin: I. Physico-chemical properties and some structural aspects of camel hemoglobin ( Camelus dromedarius)

Hemoglobin from an adult camel ( Camelus dromedarius) was prepared from the red cell lysate by CM-and DEAE-cellulose chromatography. The purified hemoglobin showed a lesser mobility on starch gel electrophoresis at pH 8.5 than that of human hemoglobin C. Native camel hemoglobin contains 95–99% alkali-resistant hemoglobin and is soluble in2.94 M K 2HPO 4/KH 2PO 4 buffer. Different forms of camel hemoglobin show similar ammonium sulfate precipitation curves. Indirect evidence for the stability of camel hemoglobin solutions was obtained from several sources. Spontaneous methemoglobin formation is extremely slow and minimal quantities of degradation products appear on starch gel electrophoresis and on chromatographic separation. The α and β chains of camel hemoglobin A were separated on a CM-23 column by the use of a pyridine formate gradient. Large peptide fragments were obtained by tryptic digestion of maleylated α and β chains. The N-terminal structure of the α and β chains and of tryptic maleylated peptides derived from α and β chains are presented. Between adult camel hemoglobin and adult human hemoglobin six amino acid differences were found in the N-terminal 20 amino acid residues of the α chain, at residues: 4, 5, 12, 14, 17, and 19; eight amino acid substitutions were found in the β chain at positions:4, 5, 6, 9, 12, 13, 16, and 19. Substitutions at α5 Ala → Lys, and β19 Asn → Lys, increase the net positive charge of camel hemoglobin by two, while other substitutions result in no charge differences. The molecular basis of the stability of camel adult hemoglobin is discussed.

Sickle Hemoglobin in Combination With

Abstract Several members of a black family from Southern Illinois were found to be heterozygous for HB JBangkok (αA2β56 gly→asp2), a hemoglobin abnormality previously described only in individuals of Thai, Chinese, or Indonesian ancestry. In two children (ages 3 and 8) Hb JBangkok was present in combination with sickle hemoglobin. Neither of these children demonstrated evidence of hemolytic disease, enlargement of the liver or spleen, or symptomatic sickle crises. The Hb J comprised 54%-59% of the total in all family members having this variant, in common with previous reports of this hemoglobin. Hb A2 and alkali-resistant hemoglobin were present in normal concentrations in all of the family members studied. Deoxygenated mixtures of Hbs S and JBangkok exhibited minimum gelation concentrations similar to those of equivalent mixtures of Hbs S and A.

Alkali resistant haemoglobin F level from birth to 12 years.

Blood samples from 652 randomly selected infants and children at different ages were examined for their HbF count by alkali denaturation method. The average foetal haemoglobin in the newborns was 61.62 per cent range (40.0 to 80.30 per cent), 40.66 per cent at the end of the 1st month of age. Less than 2 per cent was found beyond one year of age. Only traces of HbF were observed after 2 years of age. Alkali denaturation method was found to be satisfactory for the detection of HbF.

The prevalence of the rarer inherited haemoglobin defects in adult Jamaicans.

Summary. The prevalence of several haemoglobin defects, including the traits for β‐thalassaemia (0.8%), hereditary persistence of foetal haemoglobin (0.2%) and the abnormal delta chain haemoglobin, Hb B2 (2.4%), were determined from the combined results of surveys conducted on adults in a suburban and a rural community. Mean Hb A2 levels of 2.6 ± 0.4%, 5.3 ± 0.5% and 2.0 ± 0.2% were found in 639 Hb A homozygotes, seven β‐thalassaemia traits and three traits for hereditary persistence of foetal haemoglobin (HPFH), respectively. Levels of alkali resistant haemoglobin (A.R. Hb) ranged from 0.6 to 7.3% in the β‐thalassaemia traits and were 21.0, 19.0 and 16.0% in the three HPFH traits; the remaining 770 subjects in whom A.R. Hb was measured had a mean value of 0.6 ± 0.6%.

Cholelithiasis in sickle cell anemia in a caucasian population

Sixty-two Saudi Arab oasis dwellers with sickle cell anemia, aged 10 to 64 years, were evaluated for cholelithiasis. Five of them had multiple gallstones, the youngest of whom was 17 years old. Only one of these had symptoms and findings strongly suggestive of cholelithiasis and obstructive jaundice, and his serum bilirubin level was higher than is usually found in patients with the hepatic crises of sickle cell anemia. The remaining four had less severe abdominal pain and jaundice but still presented potential problems in differential diagnosis. The evaluation led to the finding of gallstones in three patients. The incidence of cholelithiasis in Saudi Arabs was found to be one-sixth to one-fourth as great as that in American Negroes. Fewer patients had impaired liver function, and over-all clinical findings were less severe. Anemia was milder and reticulocytosis was less striking. The lower incidence of cholelithiasis and the milder clinical manifestations in these patients were related to an unusually high level of alkali-resistant hemoglobin which averaged more than 20 per cent. Recommendations regarding appropriate routine evaluation of patients with sickle cell anemia by cholecystography may need to be altered for populations such as these Saudi Arabs in whom the disease is of a relatively mild type.

Blood clam (Anadara inflata) hemoglobins. Partition in aqueous two-polymer phase systems and alkali denaturation.

Anadara inflata is a clam which not only has red blood cells in its hemolymph but these nucleated cells also contain two hemoglobins. Clam hemoglobins were subjected to partition in aqueous dextran-polyethylene glycol two-phase systems and their denaturation by alkali was studied. 1. The two hemoglobins, I and II, in the clam, which have, respectively, mol. wts. 34000 and 68000, but are structurally distinct, give slightly higher partition coefficients (K) than human hemoglobin A (which has the highest K of all mammalian hemoglobins studied so far). They are not, however, too different from each other at thier isoelectric points. These data reinforce the finding that hemeproteins (unlike nonhemeprotiens) give K values in two-polymer phase systems that are relatively independent of their molecular weights. 2. Isoelectric points of the two hemoglobins were determined by the simple, recently described method of ‘cross-partition’ in the two-polymer phases and found to be pH 9.0 for hemoglobin I and 5.9 for hemoglobin II. 3. Hemoglobins I and II were found to differ in their resistance to alkalai. The alkali resistant hemoglobin constitutes 60–70% of the total hemoglobin; the remainder is non-resistant to alkalai. Column chromatography of clam hemoglobins (which yields a similar 65/35 ratio) suggests that the alkalai resistant component is hemoglobin II. Hemoglobin I is, however, more resistant to heat denaturation than is hemoglobin II.

Comparative studies on the blood of monotremes and marsupials—I. Haematology

1.1. Blood from seven species of Tasmanian marsupials and one monotreme was subjected to a haematological examination and it was found that most parameters were within normal mammalian values, but considerable variation in WBC counts was noted.2.2. The haemoglobin upon electrophoresis showed only one haemoglobin band in all species but biochemically two haemoglobins were identified, one being alkali resistant.3.3. The alkali-resistant haemoglobin is considered as foetal in origin in Eutheria but it is found in high proportions in adult marsupials, up to 78 per cent and cannot be considered as foetal haemoglobin. The blood of Tachyglossus aculeatus (Monotremata) has 90 per cent alkali-resistant haemoglobin.

Incidence of abnormal haemoglobins in different ethnic groups of Indians.

1. 2455 unrelated Indian subjects of both sexes comprising of 651 North Indians and 1804 South Indians were investigated for the incidence of abnormal haemoglobins. 2. Out of 651 North Indian subjects 11 (1.7%) had abnormal haemoglobins. The genotypes of these 11 were as follows: AD-7, DD-1, AE-1, AH-1, AF-1. 3. Out of 1804 South Indian subjects 16 (0.9%) had abnormal haemoglobins. The genotypes of these 16 were as follows: AE-10, EE-2, AH-3, AD-1. 4. Altogether 7 families — 2 for D haemoglobin and 5 for E haemoglobin were investigated. There were 13 instances (including propositi) of D haemoglobin in 2 families and there were 21 instances (including propositi) of E haemoglobin in 5 families. 5. Neither the presence of D and E-haemoglobin nor haemoglobin H-trait was associated with any haematological disorder. 6. There was a case of severe anaemia (Hb — 6.2 gm%) in one Punjabi boy of 3 months with haemoglobin AF and it was associated with abnormal red cell morphology and this boy had 48% alkali resistant foetal haemoglobin.

Studies on the blood of the tasmanian devil

1. 1. Blood from male and one lactating female Sarcophilus harrisii was subjected to haematological and biochemical examination. 2. 2. High acid phosphatase and blood urea levels were found. 3. 3. Alkali resistant haemoglobin formed a substantial proportion (27–37 per cent) of the haemoglobin, and the blood exhibited remarkable clotting power. 4. 4. α 1-globulin was not identified in the blood and γ-globulin levels were high in all individuals.

Renal artery stenosis with hypertension in Fanconi's cytopenia.

A case is described of Fanconi's cytopenia associated with thrombocytopenia, skin pigmentation and leucoderma, microcephaly, narrow palpebral fissures, abnormal thumb with absent scaphoid and hypoplastic first metacarpal, impalpable radial artery, and single pelvic kidney with renal artery stenosis and hypertension. A paternal uncle of the patient had died of leukaemia. One sibling without obvious congenital anomalies had slightly lowered platelet counts. The unusual features were renal artery stenosis to a solitary kidney, thrombocytopenia alone at the time of initial presentation and normal levels of alkali-resistant haemoglobin. This is the first report of renal artery stenosis in Fanconl's anaemia or cytopenia, and only the third reported case of stenosis of the artery to a solitary kidney. The stenosis was corrected by a renal to right-common-iliac artery anastomosis, with return of the blood pressure to normal levels.

Irreversibly sickled erythrocytes: a consequence of the heterogeneous distribution of hemoglobin types in sickle-cell anemia.

THE AMOUNT OF FETAL HEMOGLOBIN (HB F) IN ERYTHROCYTES OF PATIENTS WITH SICKLE CELL ANEMIA (HB SS DISEASE) WAS MEASURED BY TWO METHODS: (a) photometry of individual cells stained for Hb F by the Kleihauer-Betke technique; and (b) chemical assay of alkali-resistant hemoglobin in cells distributed according to specific gravity by ultracentrifugation. Irreversibly sickled cells (ISC), which could be identified directly during photometry and which were found to gather in high concentration at the bottom of ultracentrifuged cell columns, contained significantly less Hb F than non-ISC. Cell content of total Hb was constant regardless of cell size, shape, or ultracentrifugal behavior: thus absolute amounts of Hb F and S varied reciprocally from cell to cell. In experiments designed to estimate age, at formation, and rate of destruction of ISC, Hb SS blood was incubated with selenomethionine-(75)Se (which labels reticulocytes) or (51)Cr (which labels erythrocytes at random) and reinfused. Sequential blood samples were separated by ultracentrifugation into fractions rich in reticulocytes, non-ISC, and ISC; and chronological changes in the specific activity of each fraction were determined. Analogous information was obtained from radioautography of sequential blood samples after reinfusion of whole blood labeled with amino acids-(3)H: this technique permitted direct visual characterization of labeled erythrocytes as ISC or non-ISC, all of which had been reticulocytes at the time of reinfusion. The transformation of non-ISC into ISC, presumably a manifestation of membrane damage, proved to begin soon after cell release from the marrow; and ISC subsequently underwent rapid removal from the circulating blood. IT IS THEREFORE APPARENT FROM THESE STUDIES THAT, IN HB SS DISEASE, RELATIVELY SMALL RECIPROCAL CHANGES IN THE AMOUNTS OF THE TWO MAJOR HEMOGLOBINS CARRY PREDICTIVE IMPORTANCE: (a) net synthesis of Hb F is least in erythroid cells destined to become ISC; and (b) these irreversibly deformed erythrocytes suffer preferential destruction.

Haemoglobin Bart's hydrops foetalis in Thailand.

SummaryHaemoglobin Bart's hydrops foetalis is prevalent in Thailand. A total of fifteen such cases were investigated. The parents were mostly Thais and Chinese. The gestation periods varied between 7 and 10 months. Thirteen were dead in utero and the other two died within 15 min. of delivery. Ten were female and five were male. All were anaemic and oedematous with enlarged and friable placentae, and had massive hepatomegaly. The average values of haematological findings are: haemoglobin concentration, 6.77 g. %; red‐cell count, 2.93 millions/mm.3; packed red‐cell volume, 40.5%; mean corpuscular volume, 140.6 micra3; mean corpuscular haemoglobin, 24.8μμg.; mean corpuscular haemoglobin concentration, 18%. Blood smears showed hypochromia, microcytosis, macrocytosis, target cells, anisocytosis, poikilocytosis, and numerous nucleated red cells. Intraerythrocytic inclusion bodies were found in a few red cells in most cases. Sickling was positive in five out of eight cases. In every case haemoglobin Bart's comprised almost all of the haemoglobin, only trace amounts of haemoglobins A+F being present. A small amount of haemoglobin H was detectable by starch gel in three cases. Average alkali‐resistant haemoglobin was 42.3%.

Alkali-resistant Hemoglobin in Sickle Cell Disease

Article1 May 1966Alkali-resistant Hemoglobin in Sickle Cell DiseaseJOHN N. BICKERS, M.D.JOHN N. BICKERS, M.D.Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-64-5-1028 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptA substantial percentage of patients with homozygous hemoglobin S disease produce detectable amounts of alkali-resistant hemoglobin* during their adult life. The alkali-resistant hemoglobin lacks the pathologic physical properties of hemoglobin S; thus, the capacity to produce alkali-resistant hemoglobin is of potential importance as a source of functionally normal hemoglobin in sickle cell disease. The recognition of factors that affect alkali-resistant hemoglobin production may provide insight into the basic mechanisms that control its synthesis.The literature contains remarkably little information on the natural history of alkali-resistant hemoglobin in sickle cell disease and even less concerning the variables that affect its production....

Comparative studies of the respiratory functions of mammalian blood. I. Gorilla, chimpanzee and orangutan

Blood samples from a chimpanzee, two gorillas and an orangutan were used for the construction of oxygen dissociation curves at pH 7.4 and 37 °C. The morphology of the erythrocytes was also studied. Hemoglobin from each species was subjected to starch block electrophoresis. The rate of hemoglobin oxidation by potassium ferricyanide was determined as was the amount of alkaliresistant hemoglobin. The glucose-6-phosphate dehydrogenase activity in the erythrocytes was measured. With regard to all these parameters the blood of these members of the family Pongidae was essentially similar to that of the human adult except that the hemoglobin of each of these animals showed a slow rate of methemoglobin formation when exposed to potassium ferricyanide. The orangutan has large red blood cells and hemoglobin which travels slowly when subjected to starch block electrophoresis.

THE RELATION OF ALKALI-RESISTANT HAEMOGLOBIN IN THALASSAEMIA AND ABNORMAL HAEMOGLOBIN SYNDROMES TO FOETAL HAEMOGLOBIN.

Alkali‐resistant haemoglobin (haemoglobin F or foetal haemoglobin) normally constitutes the major haemoglobin component of the foetus; it is also present in gradually decreasing amounts through the first year of life. In the older individual, less than I per cent of the haemoglobin is of the foetal type; normally, most of the remaining haemoglobin is the adult variety—haemoglobin A.Alkali‐resistant haemoglobin is found in greater than normal concentrations in several blood dyscrasias. In patients with thalassaemia major, it is invariably increased and frequently constitutes the major haemoglobin component (Fessas, 1959; Beaven, Ellis and White, 1961). On the other hand, in those who are homozygous for an abnormal haemoglobin, 20 per cent or less of the haemoglobin, as a rule, is alkali‐resistant and increased percentages are not invariably found (Beaven et al., 1961; Rucknagel and Neel, 1961). The abnormal haemoglobin syndromes are most characterized by the presence of electrophoretically abnormal haemoglobin(S), although alkali‐resistant haemoglobin may be present. Neither in thalassaemia major nor in thalassaemia minor, despite the frequently increased quantity of haemoglobin A2 (Fessas, 1959), have electrophoretically abnormal components been found. Both the thalassaemia and abnormal haemoglobin syndromes, have common clinical, haematological, biochemical, genetic and anthropological features. It has not seemed unlikely that thalassaemia is based on a genetically determined abnormality similar to that resulting in the abnormal haemoglobin syndromes; it is assumed, however, that a structurally modified haemoglobin is electrophoretically undetectable (Itano and Pauling, 1961).Because an electrophoretically abnormal haemoglobin is absent in thalassaemia, one might consider that the alkali‐resistant haemoglobin is, in fact, the abnormal haemoglobin. This subject has been reviewed by Beaven and Gratzer (1959) who cite abundant biochemical and immunological evidence to the contrary. Nevertheless, recent publications report biochemical and immunological differences in the foetal haemoglobin of thalassaemia compared to that from the normal newborn (Derrien, 1959; Diacono and Castay, 1959). Studies on the structure of alkali resistant haemoglobin in thalassaemia and abnormal haemoglobin syndromes, however, have not been available.Structurally, haemoglobin A is composed of a pair of α chains and a pair of β chains. In haemoglobin F, γ chains occur instead of β chains. From genetic evidence, the production of the three types of chain appears to be under the control of sets of allelic genes at separate loci; this subject has been reviewed recently (Rucknagel and Neel, 1961). The abnormal haemoglobins are products of mutant genes at the α and β chain loci. A single amino acid

Hereditary Persistence of Fetal Hemoglobin: A Study of 79 Affected Persons in 15 Negro Families in Baltimore

AbstractHereditary persistence of fetal hemoglobin is an anomaly of hemoglobin production apparently caused by a mutant gene that inhibits synthesis of hemoglobins A and A2. Alkali-resistant hemoglobin indistinguishable from hemoglobin F of umbilical cord blood is produced, presumably as a compensatory phenomenon, so that neither anemia nor hypochromia of the red cells occurs. The data summarized are compatible with the hypothesis that function of the loci of the β and δ chains of globin is wholly suppressed, quite possibly by a mutant "operator" gene affecting linked structural loci. Heterozygotes for the anomaly have high concentrations of hemoglobin F in the erythrocytes, with a remarkably uniform distribution of fetal hemoglobin throughout the red cell population. Erythrocytes of persons with the anomaly resemble adult red cells with respect to the non-hemoglobin proteins and the oxygen dissociation curve. Experience with 79 affected Negroes in Baltimore is compared with that reported by other investigators. The occasional difficulty in differentiating the anomaly from other conditions, particularly thalassemia, is emphasized.