A behavioral and neuroanatomical investigation of the lethality caused by gestational day 11-13 retinoic acid exposure

A reversed passive latex agglutination test (RPLA) using anti-canine haemoglobin (Hb) antibody was developed for detecting bleeding in the lower digestive organs in dogs, and its applicability as a simple test for faecal occult blood was assessed. In Ouchterlony's gel immunodiffusion test, the anti-canine Hb antibody used to sensitize the latex reacted with canine Hb but not with Hbs, plasmas or meat extracts from pigs, goats, sheep, cattle, horses or chickens, or with fish extracts. Using latex sensitized with 50 micrograms/mg of anti-canine Hb IgG antibody, the lowest limit of detection for canine Hb was 21 micrograms/ml, and the latex reacted negatively with all test specimens other than canine Hb. In an in vitro experiment with a mixture of canine faeces and erythrocytes, the antigenicity of the Hb was found to undergo only very slight changes even when the specimens were allowed to stand for 12 h at room temperature. Hb could not be detected by RPLA in any of four successive faecal samples from three experimental dogs after infusion of autologous blood (5, 3 or 1 ml) into the stomach. In 3 other experimental dogs given an infusion of autologous blood (5, 3 or 1 ml) into the ascending colon, the presence of Hb was confirmed by RPLA in all four successive faecal samples obtained from those which received 5 or 3 ml of blood and in all except that obtained following the first defecation from the animal which had received 1 ml of blood.     

Effects of gamma irradiation on red cells from donors with sickle cell trait

BACKGROUND: Transfusion-associated graft-versus-host disease can be prevented by gamma irradiation of blood components. Red cells (RBCs) from sickle cell disease patients may exhibit oxidative changes of RBC membranes due to the instability of hemoglobin (Hb) S. Persons with sickle cell trait are eligible to donate blood, and 35 to 45 percent of their total Hb is Hb S. The effect of gamma irradiation on RBCs from such persons is of interest. STUDY DESIGN AND METHODS: RBCs from 12 donors with sickle cell trait (Hb AS) and from 12 with normal Hb (Hb AA) were studied. Each of the 24 RBC units was divided equally into two transfer bags via a sterile connecting device. One bag from each RBC unit received a 2500-cGy dose of gamma irradiation at its mid-plane and was stored at 4 degrees C; the second set of bags was stored without irradiation. For RBCs from 6 donors with Hb AS and 6 donors with Hb AA, units were irradiated on Day 7 and studied on Day 35 of storage (Group 1). For the RBCs from the other 6 donors with Hb AS and the other 6 donors with Hb AA, units were irradiated on Day 28 and studied on Day 42 of storage (Group 2). RESULTS: For Group 1 and Group 2, plasma potassium and plasma Hb concentrations were significantly higher and RBC ATP concentrations were slightly lower in the irradiated units than in the nonirradiated units. In Group 1 and Group 2, there were no significant differences in the plasma potassium or RBC ATP concentrations in either the irradiated or the nonirradiated units of RBCs from donors with Hb AS and donors with Hb AA. Plasma Hb concentrations were consistently lower in the units from donors with Hb AS, whether or not they were irradiated. However, in both groups, proportionally similar changes in plasma Hb concentration were detected when the irradiated Hb AS and Hb AA units were compared to nonirradiated Hb AS and Hb AA units. CONCLUSION: Gamma irradiation of RBCs from donors with Hb AS or with Hb AA resulted in comparable changes in plasma potassium, RBC ATP, and plasma Hb concentrations, although donors with Hb AS had lower plasma Hb. RBCs from donors with Hb AS subjected to 2500 cGy of gamma irradiation did not evidence a storage lesion greater than that seen in RBCs from donors with Hb AA.     

Internalization of G protein-coupled receptors in single olfactory receptor neurons

The cardiovascular effects of human albumin (Alb) and three human hemoglobin (Hb) solutions, dextran-benzene-tetracarboxylate Hb, alphaalpha-crosslinked Hb, and o-raffinose-polymerized Hb were compared in anesthetized rabbits undergoing acute isovolemic hemodilution with Hct reduction from 41.4 +/- 2.7 to 28.8 +/- 1.6%. The impact of the vasoconstricting properties of Hb was examined by measuring heart rate (HR), mean arterial pressure (MAP), abdominal aortic, and femoral arterial blood flow, vascular resistance (VR), and aortic distension during the first 3 h after hemodilution. The impact of the hemorheological parameters was assessed by measurements of hemodiluted blood viscosity. In contrast to Alb, the Hb solutions elicited an immediate increase in MAP (20-38%). The effects of Alb and Hb solutions on HR, as well as on aortic and femoral arterial blood flow, were similar. VR decreased with Alb (20-28%) and increased with all three Hb solutions (30-90%), but the MAP and VR rising trends were different with each Hb solution. Aortic distension decreased in Hb groups compared with the Alb group for the first 60 min. The viscosity of hemodiluted blood was similar for all groups at high shear rates but was dependent on the viscosity of the solutions at low shear rates. We conclude that the vasoconstriction elicited by the Hb solutions overrides the vasodilation associated with viscosity changes due to hemodilution and would be the major factor responsible to the cardiovascular changes.     

Hemoglobin and albumin adducts of benzene oxide among workers exposed to high levels of benzene

Benzene oxide (BO) reacts with cysteinyl residues in hemoglobin (Hb) and albumin (Alb) to form protein adducts (BO-Hb and BO-Alb), which are presumed to be specific biomarkers of exposure to benzene. We analyzed BO-Hb in 43 exposed workers and 42 unexposed controls, and BO-Alb in a subsample consisting of 19 workers and 19 controls from Shanghai, China, as part of a larger cross-sectional study of benzene biomarkers. The adducts were analyzed by gas chromatography-mass spectrometry following reaction of the protein with trifluoroacetic anhydride and methanesulfonic acid. When subjects were divided into controls (n = 42) and workers exposed to < or =31 (n = 21) and >31 p.p.m. (n = 22) benzene, median BO-Hb levels were 32.0, 46.7 and 129 pmol/g globin, respectively (correlation with exposure: Spearman r = 0.67, P < 0.0001). To our knowledge, these results represent the first observation in humans that BO-Hb levels are significantly correlated with benzene exposure. Median BO-Alb levels in these 3 groups were 103 (n = 19), 351 (n = 7) and 2010 (n = 12) pmol/g Alb, respectively, also reflecting a significant correlation with exposure (Spearman r = 0.90, P < 0.0001). The blood dose of BO predicted from both Hb and Alb adducts was very similar. These results clearly affirm the use of both Hb and Alb adducts of BO as biomarkers of exposure to high levels of benzene. As part of our investigation of the background levels of BO-Hb and BO-Alb found in unexposed persons, we analyzed recombinant human Hb and Alb for BO adducts. Significant levels of both BO-Hb (19.7 pmol/g) and BO-Alb (41.9 pmol/g) were detected, suggesting that portions of the observed background adducts reflect an artifact of the assay, while other portions are indicative of either unknown exposures or endogenous production of adducts.     

Evaluation of automated methods of measuring hemoglobin and hematocrit in horses

OBJECTIVE: To evaluate the accuracy of 3 automated methods of determining Hct and hemoglobin (Hb) concentration, compared with manual methods. Animals-22 clinically normal adult horses of various breeds. PROCEDURE: A blood sample was obtained from each horse. Six dilutions (representing Hct of 0, 10, 20, 40, 60, or 70%) were prepared from each sample and analyzed, using 1 of 2 blood gas analyzers or a hemoximeter (for automated determinations) or the Wintrobe macrohematocrit and cyanmethemoglobin methods (for manual determinations). Regression analysis was used to determine mean slope relationships between Hct and Hb measurements obtained by use of manual versus automated methods. Slopes were compared, using Student's t-test. RESULTS: Of the 3 automated methods examined, only 1 blood gas analyzer reported Hct and Hb values that were not significantly different from those determined by use of manual methods; however, this analyzer could not report Hb concentrations below 2.5 g/dl. The other blood gas analyzer reported values for Hct and Hb concentrations that were consistently higher than those obtained by use of manual methods at Hct < or = 20% and Hb < or = 6.6 g/dl. The hemoximeter yielded more accurate results if the Hb concentration was between 6.6 and 20 g/dl. CONCLUSION: Although there were some limitations in measuring at low Hb concentrations, the method of determining Hb concentration and Hct with blood gas analyzer 2 was more accurate than that with blood gas analyzer 1 (Hct and Hb concentration) or the hemoximeter (Hb only).     

Work capacity after iron treatment as a function of hemoglobin and iron deficiency

The relative importance of hemoglobin (Hb) and non-Hb iron for physical work capacity was studied in 45 adult male and female subjects, with a range of Hb and serum iron levels. Maximal work capacity, heart rate, venous blood lactate and serum protein were measured before and after 1 week of treatment with Imferon, i.v. Even though some non-Hb related effects on parameters indicative of maximal work capacity were found, the main factor was Hb related. Subjects with low Hb-high serum iron worked longer than ones with low Hb-low iron. When work performed was similar, the marginal Hb-low iron group had a higher blood lactate concentration than the high Hb-high iron and marginal Hb-high iron groups. The coefficient of correlation between serum iron and post-exercise lactate levels was -0.41 (p less than 0.05). Even though neither of these groups showed a Hb response within 1 week of iron treatment, the initial low serum iron groups had significantly lower heart rates at a given work load relative to subjects with high iron but with a similar Hb level. This occurred both at rest and during light to heavy exercise. These results suggest that a rather rapid benefit of iron treatment is gained in iron-deficient subjects with severe and moderate anemia which cannot be accounted for by Hb changes. Although the primary factor which affects the physical work capacity of iron-deficient anemic subjects seems to be the Hb level, there also seems to be a significant non-Hb related effect of iron treatment as well.     

Effect of hemoglobin status on humoral immune response of weanling pigs differing in coping styles

The effects of hemoglobin (Hb) status and coping style of pigs on performance and humoral immune response were studied. Twenty-four, 4-wk-old crossbred barrows were assigned to groups of three pigs based on weight and litter origin. Groups were allotted according to a 2 x 2 factorial treatment arrangement: two blood Hb concentration classes (low vs high) and two immunization procedures (control vs immunized). Immunized pigs received an antigen cocktail containing keyhole limpet hemocyanin (KLH), ovalbumin (OA), and tetanus toxoid (TT) at weaning. Additionally, pigs were stratified according to behavioral coping style in response to exposure to a stressor. During 41 d after weaning (approximate time of immunization), blood Hb concentration, ADG, and ADFI were measured weekly and serum antibody titers to KLH, OA, and TT twice weekly. Average Hb concentration differed between low and high Hb pigs (P < .001; 10.0 vs 12.0 g/dL), but this difference declined with time after weaning. Neither immunization procedure nor coping style affected Hb concentrations. In addition, ADG and ADFI were unaffected by any of the treatments. However, ADG was slightly greater in high Hb status pigs (586 vs 633 g/d) and was paralleled by a slightly greater ADFI in high Hb status pigs (812 vs 899 g/d). Antibody responses were negatively or not related to Hb status at weaning. Antibody responses (depending on isotype and antigen) were or tended to be lower in pigs with high blood Hb concentrations. Behavioral coping style strongly affected humoral immune responsiveness; enhanced or accelerated antibody responses were found in pigs that had a passive coping style.     

Polymorphisms of the human beta-defensin-1 gene

Erythropoietin (EPO) plasma levels were monitored during the perioperative period in 61 consecutive patients (22 males - 39 females), aged 62.5 +/- 9.5 years, scheduled for hip arthroplasty. All patients underwent intraoperative blood salvage (IOBS) and were subdivided into three different groups according to their hemoglobin levels (Hb) 24 hours postoperatively (group A: Hb < 8 g/dl; group B: Hb between 8-9 g/dl; group C: HB > or = 9 g/dl). Seventy-two hours after surgery EPO levels were significantly different in group A (135 +/- 68) compared to group C (54.3 +/- 32), with a positive correlation (p < 0.01) between Hb and EPO levels. On the basis of these results we suggest that a programmed autologous red blood cell collection aimed at obtaining the lowest hemoglobin values during the first 24 hours after surgery, may be of clinical utility in preventing homologous blood needs.     

Passing of gallstones via the urethra: a complication of laparoscopic cholecystectomy

The phenotype of high K (HK) red blood cells, which is an autosomal recessive, was found in dog groups from 10 of 13 breeds or populations in Japan. The incidence of HK was 26 to 38% in the San'in-Shiba, Shinshu-Shiba and Akita breeds, and the gene frequencies of HK ranged from 0.513 to 0.612. The highest incidence (42%) was found in the Jindo breed from Korea, and the gene frequency was 0.652. Two other groups from Korea also possessed this HK variation. However, although HK cells were not found in dogs from Taiwan, Indonesia, Mongolia and Sakhalin, Russia, the HK phenotype is clearly distributed now throughout Japan and Korea.     

Hb Malm? [beta-97(FG-4)His-->Gln] leading to polycythemia in a Dutch family

We have examined six individuals from a two-generation Dutch family for a suspected hemoglobin (Hb) abnormality. The propositus presented with polycythemia and complained of persistent weakness, headache, and epistaxis. All family members initially showed a normal Hb-electrophoretic pattern, but on isoelectric focusing, three of them displayed a fast-moving band associated with high packed red cell volumes (PCV) and increased red blood cell count. The Hb mutant was analyzed at the DNA level by specific gene fragment amplification (PCR), followed by direct DNA sequencing, and the mutation was confirmed by restriction enzyme analysis. We found a C-->G transversion (CAC-->CAG) at codon 97 of the beta-chain, which corresponded to the His-->Gln amino acid substitution previously described as Hb Malm?. We report here the clinical history of the patient, the effects of phlebotomy treatment, and the effect of subnormal iron conditions on the erythropoietic recovery after phlebotomy. The mechanism responsible for the induction of the higher oxygen affinity is discussed, as are some aspects concerning the occurrence, pathology treatment, and the genetic risk of Hb variants with high O2 affinity.     

Insulin-like growth factor I and insulin-like growth factor binding protein 3 as determinants of blood hemoglobin concentration in healthy subjects

We studied the serum concentrations of IGF-I, IGF-binding protein 3 (IGFBP-3), and testosterone in relation to blood Hb in 60 healthy prepubertal or early pubertal boys twice, with a 9-mo interval. Serum IGF-I and testosterone levels were measured by RIA, and serum IGFBP-3 was measured by monoclonal immunofluorometric assay. Positive correlations were observed between the concentrations of blood Hb and serum IGF-I at the first examination (r = 0.36, p = 0.008) and Hb and IGFBP-3 at both examinations (r = 0.53, p < 0.001, and r = 0.39, p = 0.003). No association between Hb and testosterone concentrations was found. Our results show that blood Hb is positively correlated to serum IGF-I and IGFBP-3 levels, indicating indirectly the involvement of growth hormone in the regulation of physiologic Hb concentration. Because no association was found between Hb and testosterone concentrations, this may indicate that the role of androgens in erythropoiesis may be different at different stages of puberty. It is concluded that the IGF system may be involved in the rise of Hb level during early puberty.     

Genetic variation within the Hereford breed of cattle

Genetic differentiation among Hereford populations from Britain, Ireland, Sweden, Canada and New Zealand together with six other beef breeds was assessed using blood type polymorphisms. Changes in the genetic structure of the British Hereford population over time were also examined. Loci surveyed were seven red cell antigen systems (A, B, C, F, L, S, Z), and two serum protein loci (transferrin and albumin). Within group variation was measured by the average expected heterozygosity, and between group relationships by genetic distance. There was significant genetic differentiation among Hereford populations from different countries. Differences between Hereford groups, however, were not as large as differences between breeds. There were also significance differences among British herds. The proportion of Canadian genes in the British 'hybrid' population was estimated to have increased from 0.42 (+/- 0.34) in the 1970s to 0.98 (+/- 0.11) in the 1990s. Canadian Hereford groups were found to be less heterozygous than other groups, and replacement of the British population with Canadian animals may lead to loss of variation. Breeding strategies that preserve original native genes in British Hereford populations should be considered by commercial breeders, in order to prevent the long-term loss of genetic variation within the breed.     

The i-STAT analyzer. A new, hand-held device for the bedside determination of hematocrit, blood gases, and electrolytes

Exact and quick measurements of basic laboratory parameters are important in selected patients in the perioperative period. Depending on the capabilities of a hospital's central laboratory, the anaesthesiologist may only obtain such laboratory tests after unacceptable delays. This problem may be overcome by a new bedside measurement device that has become available from i-STAT Corporation, Princeton, USA. The hand-held, battery-driven analyser accepts blood specimens that are injected into a disposable cartridge (EG7+) and measures acidity, blood gas tensions, haematocrit, and electrolytes. The aim of this study was to determine the accuracy of such measurements by comparing them with measurements obtained by conventional laboratory test methods. METHODS: Heparinised arterial blood specimens were collected in duplicate from 49 surgical patients. Measurements of ionised calcium (Ca), sodium (Na), potassium (K), pH, pCO2, pO2, base excess (BE), haematocrit (Hct), and haemoglobin (Hb) obtained by the i-STAT analyser were compared with measurements from the calibrated analysers ABL 615 and EML 100 (Radiometer, Copenhagen). Because the i-STAT analyser calculates the Hb concentration from a conductometrically measured Hct, 19 blood specimens were centrifuged in order to compare test results with conventionally obtained Hct and Hb values. As the Hct test sensitivity with the i-STAT changes with diluted blood due to its low albumin concentration, Hct and Hb measurements during cardio-pulmonary bypass (CPB) must be corrected by activating an analyser-implemented correction algorithm (Hct/CPB and Hb/CPB). Correlation analysis was performed between conventional measurements and i-STAT values (Ca, Na, K, Hct, pCO2, pO2), between values that the i-STAT analyser derives (Hb, HCO3, BE) and conventionally obtained results, and between normal and CPB-corrected Hct and Hb values. Accuracy was judged according to the national quality standard, which requires test results to lie within the 95% confidence interval of conventional tests. RESULTS: Each blood specimen was analysed: erroneous results or technical failures did not occur. Measurement of one set of i-STAT values required 2.5 min. Correlation coefficients (r) between conventional and i-STAT results were: 0.85 for CA, 1.0 for K; 0.86 for Na; 0.99 for pH; 0.98 for pCO2; 0.99 for pO2; 0.93 for HCO3; 0.93 for BE; 0.46 for Hb values not corrected for CPB and 0.95 for CPB-corrected Hb; and 0.74 for Hct values not corrected for CPB and 0.98 for CPB-corrected Hct. The correlation coefficient for Hct between centrifuged and CPB-uncorrected i-STAT values was 0.81 and that for CPB-corrected values was 0.98. National accuracy requirements were not met for tests of: Ca (by 0.02 mmol/l); pH (by 0.01); pO2 including hyperoxic values (by 26.7 mmHg, but were met for pO2 values < 200 mmHg); Hb (by 1.6 g/dl); Hb/CPB (by 0.8 g/dl); and Hct (by 6.5%, but were met for Hct/CPB values). All other tests fulfilled the required standards. CONCLUSION: This analyser is easy to use, reliable, and portable, and therefore suitable for the operating room, for analyses during emergencies, on peripheral wards, for preclinical screening, or at times when availability of lab tests is time-consuming or limited. The test accuracy for electrolytes, blood gases, and Hb is high enough to justify routine use of the i-STAT analyser in clinical practice. That the nationally required quality standards for Ca, pH, and Hb were not met is not of importance because the measured deviation was too small to have clinical relevance. When analysing diluted blood with a low Hct and low oncotic pressure, it is important to activate the analyser's correction algorithm "CPB", because the obtained results will then comply with the required accuracy.     

Hb Helsinki: a variant with a high oxygen affinity and a substitution at a 2,3-DPG binding site (beta82[EF6] Lys replaced by Met)

A new haemoglobin, Hb Helsinki, in which beta 82-Lys (EF6) is replaced by Met, was found in a Finnish family. It was associated with familial erythrocytosis, and the oxygen affinity of the blood was higher than normal. The oxygen equilibrium curves of purified Hb Helsinki and HbA from the same haemolysate have been determined under vaious conditions. "Stripped' Hb Helsinki was found to show normal cooperativity, slightly low oxygen affinity and a reduced Bohr effect at physiological pH. However, the organic phosphates, 2,3-diphosphoglycerate (2,3-DPG) and inositol hexaphosphate (IHP) had a very small effect on Hb Helsinki, and the 2,3-DPG binding constant of deoxygenated Hb Helsinki is close to that of oxyhaemoglobin A. Thus, the replacement of Lys by Met at position 82 dramatically changes the nature of the central cavity of the tetramer and the effect of 2,3-DPG on the respiratory function of the molecule.     

Identifying the conformational state of bi-liganded haemoglobin

While most researchers agree on the global features of cooperative ligand binding to haemoglobin (Hb), the internal mechanisms remain open to debate. This is not due to inaccurate measurements, but is rather a consequence of the cooperative ligand binding that decreases the equilibrium populations of the partially liganded states and makes observation of the transitions between these substates more difficult. For example, the equilibrium population of the doubly liganded tetramers is typically less than 5% of the total Hb. As a result many models with widely varying mechanisms may fit the oxygen equilibrium curve, but may not be consistent with observations of other parameters, such as ligand-binding kinetics or subunit association equilibria. The wide range of methods and models has led to divergent conclusions about the properties of specific substates. One notable debate concerns the properties of the doubly liganded forms. The simple two-state model predicts a shift in the allosteric equilibrium based on the number of ligands bound, but not on their distribution within the tetramer. From studies of dimer-tetramer equilibria of various pure and hybrid forms, it was concluded that a tetramer with two ligands bound on the same alpha beta dimer (species 21, an asymmetric hybrid) shows an enhanced tetramer stability, similar to singly liganded Hb, relative to the other three types of doubly liganded tetramers which resemble the triply liganded forms [Ackers et al. (1992). Science 255: 54-63]. The implications of this model and the relevant experiments will be reviewed here.     

Effect of alpha(alpha)-cross-linked hemoglobin and pyridoxalated hemoglobin polyoxyethylene conjugate solutions on gastrointestinal regional perfusion in hemorrhagic shock

BACKGROUND: Hemoglobin-based blood substitutes may cause vasoconstriction, which could limit organ perfusion during trauma resuscitation. We investigated the effect of two hemoglobin solutions on regional blood flow and mucosal perfusion in the gastrointestinal tract in a hemorrhagic shock model. METHODS: Twenty-four swine were bled 30% of blood volume over 1 hour. Six additional animals were anesthetized and monitored but did not undergo hemorrhage. Bled animals were resuscitated with alpha(alpha)-hemoglobin (alpha(alpha)Hb), pyridoxalated hemoglobin polyoxyethylene conjugate (PHP), shed blood, or lactated Ringer's solution. Regional blood flow was measured by radiolabeled microspheres. Gastric mucosal perfusion was estimated by measuring intramucosal pH (pHi) by tonometry. RESULTS: PHP and shed blood restored small-bowel flows to sham values, whereas lactated Ringer's solution and alpha(alpha)Hb did not. Shed blood and PHP, but not alpha(alpha)Hb, restored cardiac index (CI) to baseline (p < 0.05). Mean pulmonary artery pressure was elevated over baseline with alpha(alpha)Hb and PHP and remained elevated with alpha(alpha)Hb (p < 0.05). pHi was significantly lower after resuscitation with PHP than with other fluids. CONCLUSION: PHP was efficacious in restoring CI and small-bowel flow, but the pHi remained low, indicating possible continued mucosal ischemia. Alpha(alpha)Hb led to limited recovery of CI and small-bowel blood flow but restored pHi close to baseline. Shed blood was efficacious in restoration of pHi, gastrointestinal blood flows, and systemic hemodynamics.     

Spanish study of quality of life in migraine (I). Profile of the patient with migraine attending neurology clinics

This study reports our experience with 67 intrauterine transfusions (IUTs) carried out for 27 cases of severe Rh alloimmunization, which could be useful to other developing countries with similar situations. Most of the mothers were from sections of India other than Mumbai, their socioeconomic status was low, and they were referred during the second or third trimester. The mean gestation age at first IUT was 27+/-2.9 weeks and maternal anti-D titer ranged from 1:32 to 1:512. Ultrasonography (USG) was normal in eight cases, but showed minimal or gross ascites in 8 and 11 cases, respectively. The mean +/- SD hematocrit (HCT) in three groups defined by USG was 23.5+/-1.7, 15.9+/-4, and 12+/-5.9, respectively. Amniotic fluid analysis, which proved to be an important investigation, indicated IUT in eight cases having normal USG. Six cases were severely anemic (Hb deficit >7 g/dl). By fetal cell staining, the percentage of the donor's red cells in the fetal circulation was determined. Besides Hb, blood group, direct antiglobulin test, and mean cell volume, this parameter was also useful in assessing efficacy of IUT and the need for an exchange transfusion after birth. Of 11 fetuses having gross ascites, eight and one each from the remaining two groups, were stillborn. One death may be procedure related. Two neonates died due to hemorrhagic disorder and prematurity. The overall survival rate was 55.6%. Late referral, severe Rh alloimmunization, volume overload, delay in IUT because of nonavailability of blood and use of nonirradiated blood could be the reasons for the poor outcome. Strategies for improving results are discussed.     

Serum p53 antibodies in patients with oral lesions: correlation with p53/HSP70 complexes

In order to evaluate the role of insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding protein-3 (IGFBP-3) in the maintenance of blood Hb concentration in infants, we studied the serum concentrations of IGF-I and IGFBP-3 in relation to blood hemoglobin values in 25 healthy term infants at birth and two months of age. The mean concentration of IGF-I was 18.6+/-7.1 ng/ml and IGFBP-3 was 1240+/-498 ng/ml at birth. Positive correlation was observed between the blood Hb concentrations and both IGF-I (r = 0.56, p = 0.004) and IGFBP-3 levels (r = 0.38, p = 0.04) at the first examination. Our results show that blood Hb is positively correlated to serum IGF-I levels indicating indirectly the involvement of mediators of growth hormone in the regulation of physiologic Hb concentrations at birth. As no relationship was found between IGF-I, IGFBP-3 and Hb levels at the second examination, the same association could not be demonstrated at two months of age.     

Hemoglobin Rambam (beta69[E13]Gly-->Asp), a pitfall in the assessment of diabetic control: characterization by electrospray mass spectrometry and HPLC

Hemoglobin (Hb) Rambam, or beta69[E13]Gly-->Asp, has been identified in a German woman also suffering from non-insulin-dependent diabetes mellitus and chronic obstructive pulmonary disease. This is the first observation of this Hb variant in a German family thus far. The detailed evaluation of its structure using electrospray mass spectrometry revealed new minor glycohemoglobin components and showed that the attachment of glucose to the beta NH2 terminus occurred at an almost identical rate in both wild-type and mutant beta-chains. However, the introduction of a carboxyl group at beta69 seems to increase the glycation of epsilon-amino groups of lysine residues. The glycemic state in the propositus was well reflected by the total glycohemoglobin concentrations but not by the Hb A1c values, which did not reflect hemoglobin glycation in this patient. This case demonstrates that Hb A1c cannot be used reliably in the management of diabetic patients carrying Hb variants such as Hb Rambam. Functional studies of the whole blood of the heterozygous carrier demonstrated extremely low oxygen affinity, which may have been caused by increased 2,3-diphosphoglycerate related to chronic obstructive pulmonary disease and hyperthyroidism. None of the clinical symptoms could be directly associated to Hb Rambam.     

Association of hemoglobin Saint Etienne (alpha2beta295F8 His replaced by G1n) with hemoglobins A and F. Synthesis and subunit exchange in vitro

The unstable hemoglobin (Hb) Saint Etienne (alpha2beta295F8 His replaced by G1n) (betaSE) was found in the red blood cells of an 8-year-old boy. The composition of this hemoglobin was 26% Saint Etienne, 52% A, 3% A2 and 19% HbF. Studies of hemoglobin synthesis indicate: a) a balanced synthesis of alpha and non-alpha chains (alpha=betaA + betaSE + gamma), b) an increased pool of free alpha hemoglobin chains, and c) a rapid exchange of alpha chains between this pool and HbSE. The alpha chain pool resulted from the dissociation of HbSE and the greater instability of betaSE chains than alpha chains upon heating. Hemoglobin F is of the fetal type and is heterogeneously distributed among the red cells. Furthermore, two populations of red blood cells could be separated according to their i antigen content. Analysis of the hemoglobins revealed a heterogeneous distribution. Thus, F hemoglobin was preferentially associated with cells having low i antigen level, while Saint Etienne hemoglobin was increased in cells having a high i antigen level. HbF and HbSE were not present in the parents of the propositus. Study of the genetic markers confirmed the filiation. The parents were normal upon clinical and hematological examination; they exhibited a normal pattern and synthesis of hemoglobin. The Hb Saint Etienne case is compared with Hb Istanbul, which in spite of the same amino acid substitution is not associated with increased HbF level.