Predictive value of cord blood hematological indices and hemoglobin Barts for the detection of heterozygous alpha-thalassemia-2 in an African-Caribbean population. (9/584)

BACKGROUND: Cord blood hemoglobin Barts (HbBarts) and hemocytometric indices may be used for classification of newborns into those without alpha-thalassemia-2 (alphaalpha/alphaalpha) and with heterozygous alpha-thalassemia-2 (-alpha(3.7)/alphaalpha). We investigated by logistic regression analysis whether the combination of HbBarts and hemocytometric indices improves classification compared with classification based on a single analyte. METHODS: HbBarts percentages and hemocytometric indices were determined in cord blood of 208 consecutive newborns in Curacao (Netherlands Antilles). Of these, 157 had alphaalpha/alphaalpha and 51 had -alpha(3.7)/alphaalpha, as established by DNA analysis. RESULTS: Between-group differences were significant for erythrocytes, mean cell volume, mean cell hemoglobin (MCH), mean cell hemoglobin concentration, platelets, hemoglobin F(0) (HbF(0)), and HbBarts. The Logit equation of the logistic regression model, using MCH (pg) and HbBarts (%), was: 42.7164 + 5.7916(HbBarts) - 1.3110(MCH). A sensitivity of 100% was reached at a Logit value of -3.70. The corresponding specificity was 62.2%, and the predictive value of a positive test (PV+) was 46.3% (95% confidence interval, 37.0-55.7%). The relative information gains were as follows: 88% for the HbBarts-MCH combination, 26% for MCH (not significant), and 0% for HbBarts compared with the 24.6% -alpha(3.7)/alphaalpha prevalence. CONCLUSION: Combined use of cord blood HbBarts and MCH improves classification compared with classification based on single hemocytometric indices.  (+info)

Hb Ube-2 in a diabetic case with an abnormally low HbA1C value. (10/584)

A 69-year-old male diabetic patient had an abnormally low HbA1C value of 2.8%, which was inconsistent with his elevated fasting plasma glucose of 8.2 mmol/l. Hb analysis disclosed that the abnormal Hb was Hb Ube-2 [alpha68 (E17) Asn --> Asp] and it accounted for 21.5% of the total Hb. Since the glycated abnormal Hb emerged at the same position as did HbF on high performance liquid chromatography, the HbA1C value was falsely low. The present case demonstrates that Hb Ube-2 is one of the abnormal Hbs in which caution should be exercised when monitoring diabetic control.  (+info)

The role of amino acid alpha38 in the control of oxygen binding to human adult and embryonic haemoglobin Portland. (11/584)

The role of the amino acid at position alpha(38) in haemoglobin has been probed using site-directed mutagenesis. When the Thr residue at position alpha(38) (which is totally conserved in all mammals) is changed to a Gln, the equilibrium properties of the protein are significantly altered. Equilibrium and kinetic data show that the R-state properties of the protein are essentially unaffected by the mutation whilst the allosteric equilibrium and T-state properties are changed. Mutation of the naturally occurring Gln(38) of the human embryonic haemoglobin zeta-chain (the only known non-Thr containing globin) to a Thr residue shows the converse change in properties produced by the adult mutation, although in this case the situation is complicated by significant chain heterogeneity in the T state. An extension of the two-state model of co-operativity is presented to describe quantitatively the equilibrium ligand binding in the presence of T-state chain heterogeneity. A molecular model is described in which the putative interaction of alphaGln(38) and betaTyr(145) is identified which make a significant contribution to the previously reported unusual ligand-binding properties of the zeta-chain containing human embryonic haemoglobins.  (+info)

Hemoglobin interaction in sickle cell fibers. I: Theoretical approaches to the molecular contacts. (12/584)

Computerized molecular model building has been used to deduce the arrangement of sickle cell hemoglobin molecules (Hb-S) in the tubular fibers which form within sickling cells and in concentrated cell-free solutions of deoxygenated Hb-S. A "best" solution has been found which satisfies all of the reported properties of these fibers. In the proposed arrangement the contact between adjacent Hb-S molecules in the direction parallel to the fiber axis is primarily hydrophobic and in addition contains two salt bridges between the molecules. This contact would be disrupted with the Glu of Hb-A at the beta6 position instead of the Val of Hb-S, and it would not make a long fiber with oxygenated Hb-S. Residues in the A helix and the GH corner of the beta2 chain of one molecule are in contact with residues of the A, B, and E helices and the GH corner of the alpha1 chain of its neighbor. The intermolecular contact in the direction perpendicular to the fiber axis is mainly between the end of the E helix and the EF corner of the beta1 chain on the first molecule and the F helix and FG corner of the alpha2 chain of its neighbor. Some of the implications of these contacts are reported here, and others will be presented in subsequent papers.  (+info)

Identification of the Hb Lepore phenotype by HPLC. (13/584)

BACKGROUND AND OBJECTIVE: Hb Lepore is a structurally abnormal hemoglobin in which the abnormal globin chain is a hybrid or fused globin chain (db). Three different Lepore hemoglobins have been identified, differing from each other in the point at which the db fusion occurs; Hb Lepore Hollandia (d22/b50), Hb Lepore Baltimore (d59/b86) and Hb Lepore Boston (d87/b116). In Spain only Hb Lepore Boston and Hb Lepore Baltimore have been identified. Hb Lepore is easily detected by electrophoretic and chromatographic studies, whereas the type of Hb Lepore is distinguished by chromatography of tryptic peptides of abnormal db chain. In this work, we show an easier chromatography technique for identifying the Hb Lepore phenotype. DESIGN AND METHODS: Thirteen different unrelated families (23 patients) from different parts of Spain were studied. The existence of Hb Lepore was diagnosed by standard methodology and quantified by ionic exchange HPLC. The globin chains were studied by reversed phase HPLC, which showed us the phenotype of Hb Lepore; this phenotype was corroborated by a gold standard test using molecular biology techniques. The statistical analysis was designed to determine the behavior of the quantitative (hematologic) variables using the independent variable (Hb Lepore Baltimore or Hb Lepore Boston) categorized by Student's t-test for independent groups. The distribution of the variable was established using theoretical models and the variance homogeneity hypothesis was tested. The validity of the hematologic data was estimated by creating a receiver operating characteristic (ROC) curve. RESULTS: In the study of globin chains by reversed phase HPLC, in 14 patients (8 families) three peaks were eluted; they corresponded to a, b and db globin chains. In these cases when DNA was studied by PCR followed by digestion with the restriction enzyme Pvu II, the phenotype of Hb Lepore was identified as being the Boston variant, whereas in the rest of patients (9 in total), the peak identified with hybrid chain globin (db) was not present and the molecular study showed that these patients were heterozygotes for Hb Lepore Baltimore. INTERPRETATION AND CONCLUSIONS: The study of globin chains by reversed phase HPLC is sufficient to know the phenotype of Hb Lepore and thus tedious techniques such as chromatography of tryptic digestion product of db abnormal chains are not essential, a particularly important consideration in those laboratories that do not have the possibility of carrying out molecular biology studies. Neverteheless, we should continue to use a gold standard molecular biology test in cases of prenatal diagnosis because this technique is the most exact and reproducible that we have.  (+info)

Hemoglobin Cranston, an unstable variant having an elongated beta chain due to nonhomologous crossover between two normal beta chain genes. (14/584)

An asymptomatic woman was found to have a compensated hemolytic state due to an unstable hemoglobin variant, comprising 35% of the total. Peptide maps of tryptic digests of the abnormal beta chain were identical to those of beta A except that tryptic peptide 15 (Tyr-His-COOH) was absent and a new peptide was detected, containing equivalent amounts of Ser, Ile, Thr, and Lys. Its sequence was determined by manual Edman degradation. An additional hydrophobic peptide isolated by counter-current distribution contained: Asx, Ser, Ala, Tyr, 2 Phe, and 3 Leu. Its sequence was determined on an automatic solid phase sequencer. Digestion with carboxypeptidase A confirmed the C-terminal sequence. Hemoglobin Cranston has an elongated beta chain with the following structure: (see article). This variant is the first "adult" human hemoglobin known to contain isoleucine. It is likely that hemoglobin Cranston arose because of a nonhomologous crossover of two normal beta chain genes, probably during meiosis, with the insertion of two nucleotides (AG) at position 144, resulting in a frame shift. Hemoglobin Cranston provides new information on the structure of the beta chain gene as well as an explanation of both the structure and genetic basis of hemoglobin Tak, the only other elongated beta chain variant that has been described.  (+info)

Nucleotide sequences of the 3'-terminal untranslated region of messenger RNA for human beta globin chain. (15/584)

In normal messenger RNA for the human beta-globin chain, nucleotide sequences have been identified which can be matched to the amino-acid sequence of the abnormally long segment of the beta-chain of hemoglobin Cranston. The finding of these sequences strengthens the hypothesis that the betaCranston chain arose by a frameshift mutation allowing the "readthrough" of the normal termination codon and translation of usually untranslated portions of the messenger RNA for the beta-globin chain. The oligonucleotides which match the amino-acid sequence of hemoglobin Cranston provide a sequence of 36 nucleotides which follows the normal beta-chain termination codon UAA.  (+info)

Synthesis of hemoglobin AIc in normal and diabetic mice: potential model of basement membrane thickening. (16/584)

Adult diabetic mice (C57Bl/KsJ--db/db) have increased amounts of a minor hemoglobin in their peripheral blood compared to wild-type (+/+) mice. This increase is analogous to the 2-fold increase of a glycohemoglobin with similar chromatographic mobility (Hb AIc) seen in the blood of patients with diabetes mellitus. Although the exact chemical nature of human or mouse Hb AIc is unknown, both contain a sodium-borohydride-reducible linkage on the beta chain which is a presumed Schiff base between a sugar moiety and the protein. The db/db animals, which have normal amounts of mouse Hb AIc at weaning, show the increase approximately 4 weeks after the onset of the signs of diabetes. This rise is brought about by an increase in a circulating factor that determines directly or indirectly the synthesis of mouse Hb AIc as a post-synthetic modification of Hb A. Evidence for this was obtained by showing that the rate of synthesis of the modified Hb is linear for at least the first 50 days of the life of the red cell and that the rate of synthesis is dependent on the environment in which the cells circulate. Thus the rate of mouse Hb AIc synthesis in +/+ cells is greater when those cells circulate in a db/db host than when they circulate in a +/+ host. The nature of the humoral factor is unknown. If glycosylations of basement membrane proteins and hemoglobin proceed via a common mechanism, then the monitoring of Hb AIc could provide a useful model for studying the early events of basement membrane thickening.  (+info)