Accurate Measurements of Fetal Hemoglobin for Neonates with Different Gestational Ages

The purposes of this study were to examine the accuracy of fetal hemoglobin (Hb F, α₂γ₂) as quickly measured by a hemoximeter but verified by high performance liquid chromatography [HPLC, including Hb F total (Hb Ft), acetylated Hb F (Hb F₁), and non acetylated Hb F ( Hb F*)], and to predict the Hb F levels for different gestational weeks of neonates. Thirty-nine neonates of predominantly Hispanic and African American ethnicity, with gestational ages ranging from 25 to 38 weeks, were investigated. Analyses were performed on 163 blood samples that were pure neonates’ blood before the transfusion of any adult blood. Two neonates had increased Hb C [β6(A3)Glu→Lys, GAC→AAG] levels (1.67–2.79%) and one neonate whose mother drank alcohol during pregnancy, had elevated Hb A₂ levels (0.12–0.14%). After excluding these data points, the mean Hb F were overestimated by hemoximeter, 118.4 ± 8.77% vs. 92.6 ± 2.77% by HPLC (mean difference: 25.8 ± 7.71%, p = <0.001). Mean Hb F₁ was 10.5 ± 2.28%. Hb F levels decreased as gestational age increased (p <0.001 for Hb Ft and Hb F*; p = <0.05 for Hb F₁). A multivariate regression model for Hb F prediction was established with the best R². The gestational age and post birth hours in the prediction of Hb Ft was included when Hb F could be determined at the clinical settings. Future studies may be needed to account for Hb F₁ when measuring Hb F levels to assess oxygenation status in (pre term) neonates.     

Hematological findings in 375 Sicilians with Hb S trait

We evaluated hematological parameters in 375 Sicilian adults with Hb S trait: Hb S levels were 41.91 +/- 2.65% in males and 40.92 +/- 2.8% in females. RBC, MCV, PCV, MCH, MCHC and total hemoglobin levels were within the normal range. Only mean Hb A2 and Hb F levels were increased (Hb A2 = 2.78 +/- 0.2%; Hb F = 1.05 +/- 0.18%), although they remained inside the normal ranges when compared to healthy controls (Hb A2 = 2.48 +/- 0.19%; Hb F = 0.93 +/- 0.14%) (p less than 0.0005). We conclude that our population does not show the hematological abnormalities such as microcytosis and decreased Hb levels, observed in the Black, Indian, Saudi Arabian carriers, and that the presence of those abnormalities is probably related to the coexistence of alpha-thalassemia, rarely observed in Sicily.     

Pattern of HB F Level Rise During Normal Pregnancies

Fetal hemoglobin (Hb F) is the normal hemoglobin (Hb) that is present in the fetus and usually almost absent in adults. The objective of this study was to assess the changes in Hb F levels during normal pregnancy. The level of Hb F was determined in serial blood samples from women at different stages of pregnancy using cation exchange high performance liquid chromatography (HPLC) and compared to age and sex-matched controls. A significant increase (p <0.001) was observed in the level of maternal Hb F; the mean Hb F level during pregnancy was 0.71 ± 0.51%, while in the non pregnant control group it was 0.28 ± 0.35%. There was no significant difference in Hb F levels in the three trimester groups using the ANOVA test (F = 0.25). Correlation studies between the gestational age and level of Hb F showed no significant increase of Hb F with advancing pregnancy (R = ?0.053, p >0.05). The cause of the rise in Hb F is yet to be elucidated.     

Hemoglobin A2 in hyperthyroidism

Erythrocyte hemoglobin A2 (Hb A2) was quantitated in 28 hyperthyroid patients prior to antithyroid therapy and at intervals during therapy. Fetal hemoglobin (Hb F) levels and red cell mean corpuscular volume (MCV) were also monitored. Before therapy, Hb A2 was significantly elevated (mean +/- SD, 3.3 +/- 0.5%: normals, 2.5 +/- 0.3%: p less than 0.001). Hb F levels exceeded 1.0% in 10 cases. A tendency to red cell microcytosis was also observed (mean MCV 81.1 fl). To-date 10 of these subjects have had adequate studies following appropriate and sustained responses to antithyroid therapy. All showed a fall in Hb A2, usually accompanied by a rise in red cell MCV. A hematologic profile for hyperthyroidism is proposed.     

Glycosylated and acetylated hemoglobins in relation to gestational age and red cell age

Cord blood samples were obtained at delivery from women at gestational periods of 30 to 36 weeks (preterm) and from a control group at 40 weeks of gestational age (term). Total glycosylated hemoglobin (GHb) was determined by affinity chromatography and the percentages of the minor Hbs FIa+b and FIc (of total Hb F) were determined by Bio-Rex 70 chromatography in whole blood samples and in red cell populations separated by Dextran 40 density gradient centrifugation. The absolute levels and the increase due to red cell aging of GHb, Hb FIa+b and Hb FIc were significantly reduced in the preterm samples compared to the term samples indicating decreased glycosylation in preterm fetus. When the nonglycosylated Hbs from term and preterm newborns were separated by Bio-Rex 70 chromatography after removal of the GHb by affinity chromatography, the acetylated form of Hb FIc also showed an increase with red cell aging indicating posttranslational enzymatic or nonenzymatic acetylation of Hb F during the entire life span of red cells. Moreover, as with GHb, the acetylated Hb was also decreased in the preterm newborns.     

Haemoglobin A/F ratio in neonates at 7 days correlated with birth weight and estimated gestational age

Haemoglobin (Hb) A and Hb F has been determined in neonates of Afro-Caribbean and North European origin with gestational ages varying from 32 to 42 weeks. There was no difference in the distribution of Hb A/F ratios between the two groups. Only weak correlations could be established between the Hb A/F ratio and the estimated gestational age or birth weight. This would indicate that there is a considerable interindividual variation in the timing of the switching of haemoglobin synthesis from Hb F to Hb A and erythrocyte production from liver to bone marrow and of oxygen affinity of fetal blood. Thus, intra-uterine adjustments of the oxygen release capacity of haemoglobin would have to rest on biochemical mechanisms during the third trimester.     

Hydroxyurea increases hemoglobin F levels and improves the effectiveness of erythropoiesis in beta-thalassemia/hemoglobin E disease

Hydroxyurea (HU) is one of several agents that have been shown to enhance hemoglobin (Hb) F levels in patients with sickle cell disease and may be useful as a therapy for beta-globinopathies. However, limited information exists on the effects of HU in patients with thalassemia. Accordingly, we examined the hematologic effects of orally administered HU in 13 patients with beta-thalassemia/Hb E, including four patients who had been splenectomized. These patients were treated with escalating doses (final range, 10 to 20 mg/kg/d) for 5 months and were observed in the outpatient hematology clinic every 2 to 4 weeks. Complete blood counts including reticulocyte counts, amounts of Hb E and Hb F, G gamma:A gamma and alpha:non-alpha globin biosynthetic ratios were evaluated before and during treatment. Almost all patients responded with an average increase of 33% in Hb F levels, from a mean (+/- SD) of 42% +/- 11% to 56% +/- 8% (P < .0001), and a reciprocal decline in the percentage of Hb E from 59% +/- 9% to 49% +/- 8% (P < .001). Reticulocytosis was decreased from a mean (+/- SD) of 18.0% +/- 15.6% to 11.7% +/- 9.1% (P < .05); there was also a slight (10%) but statistically significant increase in hemoglobin levels and an improved balance in alpha:non-alpha globin chains ratios. The side effects were minimal in most patients, although these patients tended to tolerate a lower dose of HU before significant myelosuppression than has been our previous experience in sickle cell disease. One splenectomized patient died of sepsis during the trial. We conclude that increased Hb F production in beta-thalassemia/Hb E patients, with an improvement in the alpha:non-alpha globin ratios and, probably, the effectiveness of erythropoiesis, can be achieved using HU. Longer trials of HU in this population, including at other doses and in combination with other agents, appear warranted.     

Multicenter validation of fully automated capillary electrophoresis method for diagnosis of thalassemias and hemoglobinopathies in Thailand

Thalassemias and hemoglobinopathies are highly prevalent in Thailand and other Southeast Asian countries. Accurate and precise separation of hemoglobin types, together with reliable quantitation, are essential for differential diagnosis of these diseases. Presented in this study is a multicenter validation of a fully automated capillary electrophoresis (CE) method for hemoglobin separation and quantitation involving four reference laboratories in Thailand. Analytical performance characteristics, including precision and accuracy were compared with existing validated HPLC and LPLC methods using 412 blood samples from unrelated subjects. Coefficient of variance of Hb A2 quantitation was 1.80-2.86, 1.26-5.13 and 1.08-6.66% for within run, between run and interlaboratory comparison, respectively. Results of Hb A2 and Hb F quantitated by the CE method correlates well with those of the two comparative methods (r = 0.98-0.99). The CE method correctly determined the genotypes (thalassemias and hemoglobin variants) of all blood samples tested. The major advantage of the CE system is its ability to separate and quantitate Hb A2, Hb E, Hb F, Hb H and Hb Bart's, which are important parameters required for diagnosis of thalassemias and hemoglobinopathies.     

Prenatal diagnosis of beta-thalassemia major by high-performance liquid chromatography analysis of hemoglobins in fetal blood samples

In Thailand and adjacent countries, most of the beta-thalassemia genes are beta(0)-thalassemia mutations that prevent the production of Hb A. We propose the quantitation of the Hb A fraction in fetal blood in the mid-trimester of pregnancy by automated high performance liquid chromatography as a reasonable prenatal diagnostic method to be applied in areas with limited laboratory facilities. Forty pregnant women at risk of delivering a child with beta-thalassemia major were identified using an erythrocyte osmotic fragility test and quantitation of Hb A2. Cordocentesis was performed at the gestational age of 18-22 weeks and fetal blood was analyzed for hemoglobin fractions by automated high performance liquid chromatography. The beta-globin gene mutations were characterized by beta-globin gene sequencing. The 4 bp deletion at codons 41/42 (-TTCT) was the most frequent of the 40 beta-thalassemia mutations observed (20/40 = 50%), followed by the splice site mutation IVS-I-1 (G-->T) (7/40 = 17.5%), the nonsense mutation at codon 17 (A-->T) (7/40 = 17.5%), the nonsense mutation at codon 35 (C-->A) (3/40 = 7.5%), and the beta(+)-thalassemia promoter mutation at -28 (A-->G) (3/40 = 7.5%). High performance liquid chromatography revealed nine fetuses which had only Hb F and no Hb A. All were homozygotes or compound heterozygotes for beta(0)-thalassemia mutations. In the remaining 31 fetuses, a Hb A peak was present in the chromatograms. One fetus with 0.5% Hb A was a compound heterozygote for the -28 (A-->G) and codons 41/42 (-TTCT) mutations. In the remaining 30 fetuses, the Hb A values ranged between 0.8 and 7.4%. Twenty of these, with a Hb A concentration of 1.82 +/- 0.49% (range 0.8-2.8%), were beta-thalassemia heterozygotes. The remaining 10 fetuses had Hb A values of 4.89 +/- 1.47% (range 2.9-7.4%) and normal beta-globin genes. The absence of Hb A in homozygotes or compound heterozygotes for beta(0)-thalassemia mutations and the presence of measurable amounts of Hb A in heterozygotes and normal homozygotes, permits the diagnosis of fetuses expected to develop postnatal beta-thalassemia major.     

Hb Kenya among Luo adults and young children in malaria holoendemic Western Kenya: screened by high performance liquid chromatography and confirmed by polymerase chain reaction

Hb Kenya, a fusion hemoglobin (Hb) resulting from a crossover between the (A)gamma- and beta-globin genes, is accompanied by an increased level of fetal Hb (Hb F) in adult life. This study describes the use of cation exchange high performance liquid chromatography (HPLC) in the identification of Hb Kenya and of a polymerase chain reaction (PCR) method for confirmatory diagnosis. Data came from 584 children and 406 adults who were screened for eligibility for malaria vaccine trials at Kombewa, Western Kenya. Sixteen subjects (13 children and three adults) had elevated Hb F (5.0-28.4%; normal <5.0%). Of these, 11 had an apparent markedly elevated Hb A(2) (9.2-22.9%) and were confirmed by gap-PCR as having the 22.7 kb deletion characteristic of Hb Kenya. Of the five cases with elevated Hb F but normal A(2), none had Hb Kenya. We propose that in this population, the finding by cation exchange HPLC of an elevated Hb F (>9.0%) and of an apparently increased Hb A(2) (>9.2%), may suggest the presence of Hb Kenya. However, given the inability of differentiating Hb Kenya from a truly elevated Hb A(2) by routine cation exchange HPLC, it is imperative to confirm the Hb Kenya mutation by gap-PCR.     

Elevated umbilical cord ghrelin concentrations in small for gestational age neonates

Ghrelin has orexigenic effects. It is present in umbilical cord plasma in full-term neonates, raising the prospect that ghrelin plays a role in fetal and neonatal energy balance. We measured ghrelin in small (SGA), appropriate (AGA), and large (LGA) for gestational age neonates and evaluated whether ghrelin levels are modulated by neonatal insulin and glucose concentrations. Plasma concentrations of ghrelin, insulin, and glucose were measured in cord blood sampled at birth in 123 SGA, AGA, and LGA neonates (gestational age, 24-41 wk) born to mothers with and without diabetes. Ghrelin was detected in samples from all infants. Its concentration was 40% higher in SGA neonates (mean +/- SD, 2436 +/- 657 pg/ml) compared with AGA (1738 +/- 380) and LGA (1723 +/- 269) neonates. There was a positive correlation between ghrelin and gestational age in AGA/LGA (r = 0.23; P < 0.05) and a negative correlation in SGA (r = -0.67; P < 0.005) neonates. Therefore, the difference in ghrelin between SGA and AGA/LGA neonates decreases with advancing gestational age. Birth weight z-score, maternal hypertension, and glucose concentrations were significant determinants of ghrelin concentrations. In conclusion, SGA neonates present with higher umbilical cord ghrelin plasma concentrations than AGA/LGA neonates. Ghrelin may play a physiological role in fetal adaptation to intrauterine malnutrition.     

Neonatal platelets from cord blood and peripheral blood

The haemostatic system in neonates is different from that of adults, possibly contributing to an increased incidence of bleeding disorders, such as intracranial hemorrhage. In this study, we analyzed platelets from cord blood and peripheral blood, collected at three time points after delivery from 20 term and 37 preterm neonates as well as blood from 20 healthy adults. Platelet membrane glycoproteins (GP) were quantified and P-selectin expression and PAC-1 binding ability before and after stimulation with TRAP were analyzed by whole blood flow cytometry. We found no significant differences in neonatal platelets from cord blood and peripheral blood within the first 24h of life. Platelets from infants less than 30 weeks of gestation expressed lower levels of GP (33271+/-9381 vs. 44085+/-17287 for GPIIIa, P<0.05) and were less reactive than platelets from term newborns (4.3+/-3.3 vs. 20.1+/-11.8% PAC-1 positive platelets after stimulation with TRAP, P<0.05). A significantly lower level of GPIIb/IIIa expression on platelets from peripheral blood was seen in term newborns as well as preterm infants, compared to adults. There was only a partial enhancement in the degranulation ability (alpha-granules) (13.4+/-12.3 vs. 50.3+/-16.1% P-selectin positive platelets, P<0.05) and no significant increase for PAC-1 binding (13.6+/-10.9 vs. 15.3+/-5.9% PAC-1 positive platelets, P=0.8) during the first 12 days of life. In conclusion, we could demonstrate that neonatal platelet reactivity increases with gestational age.     

The influence of uridine diphosphate glucuronosyl transferase 1A promoter polymorphisms, beta-globin gene haplotype, co-inherited alpha-thalassemia trait and Hb F on steady-state serum bilirubin levels in sickle cell anemia

PURPOSE: Homozygosity for the (AT)7 allele of uridine diphosphate glucuronosyl transferase 1A (UGT1A1) gene polymorphism is associated with increased bilirubin levels in sickle cell anemia (SCA). In the present study, in addition to UGT1A1 promoter genotype, serum bilirubin level was related to other genetic modifiers -beta(S)-globin gene haplotype, Hb F, co-inherited alpha-thal trait, age and gender. METHODS: The patients were randomly selected from the sickle cell clinic, Medical College of Georgia. UGT1A1 promoter polymorphisms were determined using automated sequencing. Other investigations were with standard techniques. RESULTS: There were 67 SCA patients (41 males and 26 females), aged 2-44 yr (mean of 20.6 +/- 10.7). Ten (14.9%) patients were homozygous for the (AT)6 UGT1A1 allele, 35 (52.2%) were heterozygous for (AT)6 and (AT)7 alleles while 22 (32.8%) were homozygous for (AT)7. Serum bilirubin was significantly higher in the homozygous (AT)7 group (3.7 +/- 1.5, 3.8 +/- 2.3 and 5.6 +/- 2.4 mg/dL, respectively). It was also significantly higher in males than females and in patients aged >10 yr. There was a significant negative linear correlation (r = -0.304, P = 0.016) of serum bilirubin with Hb F. The beta-globin haplotype and co-existing alpha-thal trait did not have any significant influence on serum bilirubin levels. Patients on hydroxyurea were older, had lower Hb F, but higher mean serum bilirubin. The latter also was signifcantly higher among those with UGT1A1 (AT)7 homozygosity. CONCLUSIONS: Apart from UGT1A1 (AT)7 homozygosity, Hb F, age and gender are the other factors that significantly influence serum bilirubin level in SCA.     

The intrauterine environment is a strong determinant of glucose tolerance during the neonatal period,even in prematurity

The aim of this study was to determine the contribution of birth weight and gestational age to glucose tolerance in premature neonates. The study group consisted of 100 premature and/or small-for-gestational age infants. Anthropometric measurements were performed both at birth and at the time of a standardized milk feed carried out at 19.6 +/- 12.1 d (range, 1-65 d) after birth. Fasting and postprandial glucose and insulin levels were measured. Birth weight, as a proxy mirror of the intrauterine environment, was found to influence the glucose concentration following a standardized milk feed (beta = -0.46; P = 0.01 for birth weight z-score with 60-min glucose level), whereas gestational age did not. Small-for-gestational age neonates had higher 60-min insulin levels than appropriate-for-gestational age neonates (115.4 +/- 9.5 vs. 68.4 +/- 14.2; P < 0.05) despite similar glucose levels. Neonates born of mothers who were on antihypertensive treatment were smaller and had a higher insulin secretory response than neonates from normotensive mothers. Postnatal growth velocity (kilograms per day) correlated with birth weight (beta = -0.65; P < 0.0001) and insulin resistance (beta = -0.31; P = 0.0004), independently of each other. This study shows that glucose tolerance of the neonate is determined by weight attained at birth irrespective of gestational age and that maternal blood pressure may influence insulin sensitivity of the newborn. Furthermore, catch-up growth in neonates is determined by birth weight and insulin sensitivity.     

The usefulness of a rapid method for total fast hemoglobins determination in screening for diabetes control

We have used a simple and rapid method for the determination of total fast hemoglobins (HbA1a+b+c) in 102 diabetics and 36 normal controls. The method was described by Kynoch and marketed by Isolab. It proved to be useful in screening for patients with inadequate metabolic control in whom as a rule, total fast Hb values were higher than 8.5%. Mean Hb A1a+b+c value was significantly higher in the group of diabetics in comparison with normals (9.9 +/- 0.2 versus 6.9 +/- 0.8%). The diabetic patients were separated into four groups according to predetermined criteria of recent metabolic control. Even the patients considered to have a very good diabetes control during the past eight weeks, had supranormal total fast Hb values (7.7 +/- 0.2%). In the patients with good, poor and bad diabetes control, mean total fast Hb levels were respectively 9.3 +/- 0.3, 10.1 +/- 0.3 and 12.5 +/- 0.4%. In normals, there was a positive correlation between individual fasting blood glucose and total fast Hb values and in diabetics, mean blood glucose values correlated with total fast Hb levels. Hb A1a+b+c determinations also correlated with triglyceride values. We could find no significant association between high total fast Hb levels (greater than 8.5%) and the prevalence of retinopathy.     

Diagnosis of Thalassemia on Dried Blood Spot Samples by High Performance Liquid Chromatography

High performance liquid chromatography (HPLC) on fresh lysates is the standard test for identification of thalassemia. Samples in the form of dried blood spot(s) (DBS) mailed to reference laboratories where HPLC is available could be an alternative. Hemoglobin (Hb) on DBS at day 1, 7, 15 and 30 were analyzed by HPLC and compared to those analyzed from fresh liquid whole blood at day 0. A 100% consistent interpretation of β-thalassemia (β-thal) trait and β-thal/Hb E disease between liquid whole blood and DBS was observed when analyzing Hb A₂ on DBS at a level of 2.7–9.9% in conjunction with a lower MCV (<80 fL) and MCH (<27 pg) and analyzing β-thal/Hb E by using Hb E and Hb F at a level of 30–48% and >10%, respectively. Therefore, our results show that detection of thalassemia carriers using DBS is possible and is the alternative of choice in a low resource setting.     

Beta 0 thalassemia trait in Sardinia.

The red cell indices and results of globin chain synthesis in peripheral blood of obligate beta 0 thalassemia (beta 0 thal) carriers (parents of homozygous beta 0 thal children) and beta thalassemia (beta thal) carriers identified during mass screening are reported. Red cell indices were similar in obligate beta 0 carriers and in carriers diagnosed during mass screening. However there was a higher incidence of anemia in female obligate beta 0 thal carriers. In Sardinia the beta 0 thal carrier showed the usual hematological characteristics of the high Hb A2 beta thal carrier with microcytosis, hypochromia, reduced osmotic fragility; Hb F greater than 1% was found in 30% of the carriers. With MCV, MCH, osmotic fragility test (OFT) and Shine and Lal discriminant function we found 3.5%, 1.5%, 3.5% and 4.0% respectively false negatives in carrier identification. A part from one subject, all obligate carriers had elevated Hb A2 levels. The alpha/beta ratio in obligate carriers (mean +/- SD) was 1.83 +/- 0.26 (N = 30).     

Oral glucose tolerance: relationship with hemoglobin A1c

Fifty-two normal non-obese males and 77 male offspring of two diabetic parents, aged 15-72 years, were studied to identify possible reasons for discordance between the oral glucose tolerance test (OGTT) and hemoglobin A1c (Hb A1c). Subjects were classified into four study groups: group 1 (n = 83), normal OGTT and normal Hb A1c; group 2 (n = 19), normal OGTT and abnormal Hb A1c; group 3 (n = 9), abnormal OGTT and normal Hb A1c; and group 4 (n = 18), abnormal OGTT and abnormal Hb A1c. Glucose and insulin response were analyzed in each study group. Group 2 showed slightly higher mean glucose areas during the first 60 min of OGTT testing when compared with group 1 (P less than 0.01). Insulin levels, insulin areas and insulin/glucose regression coefficients on group 2 did not differ significantly from group 1 during OGTT. Group 3 showed significantly higher mean blood glucose levels than subjects in group 1 (P less than 0.0001) or group 2 (P less than 0.001), but significantly lower mean blood glucose levels than subjects in group 4 (P less than 0.04) throughout the OGTT. During the OGTT, group 3 showed marked absolute hyperinsulinism when compared with all other groups (P less than or equal to 0.002). Also, relative hyperinsulinism in group 3 was suggested by the elevated insulin/glucose regression coefficient (1.86 +/- 0.50) when compared with group 1 (1.17 +/- 0.09), group 2 (0.92 +/- 0.011) or group 4 (0.55 +/- 0.17) (P less than or equal to 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

G6PD enzyme activity in normal term Malaysian neonates and adults using a OSMMR2000-D kit with Hb normalization

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the commonest causes of neonatal jaundice in Malaysia. Screening of cord blood for G6PD deficiency by the semiquantitative fluorescent spot test (FST) is performed in Malaysia but this test can miss cases of partial G6PD deficiency. The OSMMR-D kit assay measures G6PD activity and hemoglobin (Hb) concentration, allowing direct expression of results in U/gHb. We evaluated this method and established the normal range for G6PD activity in normal term neonates and adults. EDTA blood from 94 neonates and 295 adults (age 15-59 years old) with normal Hb and FST were selected. The normal means for G6PD activity for neonates and adults were 12.43 +/- 2.28 U/gHb and 9.21 +/- 2.6 U/gHb, respectively; the reference ranges for normal G6PD activity in neonates and adults were 10.15-14.71 U/gHb and 6.61-11.81 U/gHb respectively. There were no significant differences in mean normal G6PD activity between the Malays and Chinese racial groups or between genders. The upper and lower limit cut-off points for partial deficiency in neonates were 7.4 U/gHb (60% of the normal mean) and 2.5 U/gHb (20% of the normal mean), respectively. For adults, the upper and lower limit cut-off points for partial deficiency in adults were 5.52 U/gHb (60% of the normal mean) and 1.84 U/gHb (20% of the normal mean), respectively. The quantitation of G6PD enzymes using this OSMMR-D kit with Hb normalization was simple since the Hb was analyzed simultaneously and the results were reproducible with a CV of less than 5%.     

Clinical, hematological, and molecular features in Sicilians with sickle cell disease.

We report the clinical, hematological, and molecular findings observed in 32 Sicilian patients with sickle cell disease. None of our patients received regular blood transfusions and careful infectious disease prophylaxis was carried out for all. Haplotyping of beta S chromosomes was performed in all patients; all were homozygous for haplotype #19 (Benin). Gene mapping excluded the presence of an alpha-thalassemia in 13 of our patients; none of the relatives showed any evidence of the presence of alpha-thalassemia. Hb F levels were 11.8 +/- 5.9% with G gamma representing 39.6 +/- 3.6% of total gamma chain. Hb F levels were higher in females than in males (12.5 +/- 5.9% versus 9.7 +/- 6.5%) but the difference was not statistically significant. All patients, regardless of age and sex, were anemic with normal mean corpuscular hemoglobin concentration, high mean corpuscular volume and mean corpuscular hemoglobin, and mild reticulocytosis. Analysis of clinical manifestations suggests that our patients have a disease of moderate severity.