Jong Weon Choi

Does Helicobacter pylori infection relate to iron deficiency anaemia in prepubescent children under 12 years of age

Aim: To investigate the association between Helicobacter pylori (H. pylori) infection and iron deficiency anaemia (IDA). Methods: Haemoglobin levels, iron parameters and serum IgG antibodies to H. pylori were measured in 693 children aged 9 to 12 y. Results: No significant differences in the seroprevalence of H. pylori infection and antibody titres to H. pylori were found between the IDA group and the non‐anaemic controls.

Erythropoietic activity and soluble transferrin receptor level in neonates and maternal blood

Serum transferrin receptor (sTfR) concentration reflects functional iron status and erythropoietic activity. The aims of this study were to examine gender differences of erythropoiesis in newborns and to evaluate the influences of maternal anaemia or iron deficiency on foetal cord blood parameters for iron status and sTfR. In total, 527 newborns and their mothers were examined. Reticulocytes were analysed by flow cytometry and sTfR was measured by an immunoenzymo‐metric method. There were no sex differences in haematological or iron parameters. However, the reticulocyte maturity index (RMI) of male neonates was 37.45%, significantly higher than the 26.81% in female neonates (p < 0.01). The high fluorescence reticulocytes (HFR) and middle fluorescence reticulocytes (MFR) of male neonates were 4.91% and 22.36%, respectively, while those of female neonates were 3.31% and 17.83%, respectively (p < 0.01 for each gender). The sTfR concentrations of male and female neonates were 6.27 mg/l and 5.09 mg/l, respectively (p < 0.01). Values for serum iron, ferritin and reticulocyte subpopulations were significantly lower in the newborns of anaemic mothers. However, newborns of iron‐deficient mothers showed no differences in iron parameters from those of non‐iron‐deficient mothers.

Nitric oxide production is increased in patients with rheumatoid arthritis but does not correlate with laboratory parameters of disease activity

BACKGROUND To investigate whether nitric oxide (NO) production correlates with laboratory parameters of disease activity in patients with rheumatoid arthritis (RA), we measured serum nitrates and nitrites (NOx) concentrations, rheumatoid factor (RF), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and serum iron markers in 115 RA patients. METHODS Serum NOx concentrations were determined after reduction of nitrates to nitrites using the Griess reaction. Serum transferrin receptor (sTfR) and serum ferritin were assayed using an enzyme immunoassay and a chemiluminescence method. Correlation coefficients of CRP, ESR, and RF vs. NOx concentrations were assessed. RESULTS Serum NOx concentrations averaged 208.3+/-126.8 micromol/l in RA patients without iron deficiency (n=72) and 267.9+/-148.4 micromol/l in RA patients with iron deficiency (n=43), which were significantly higher than those of healthy controls (32.7+/-14.3 micromol/l, p<0.01, respectively). There were no significant differences in mean NOx concentrations between the patients with CRP<0.7 mg/dl and with CRP>/=3.0 mg/dl, nor between the patients with ESR<15.0 mm/h and with ESR>/=40.0 mm/h. Serum NOx concentrations showed no significant correlations with CRP, ESR, and RF but exhibited an inverse correlation with serum iron markers. CONCLUSION NO production markedly increases in patients with RA; however, serum NOx concentration does not seem to be associated with disease activity of RA, at least on the basis of laboratory data of CRP, ESR, and RF.

Association between blood lead concentrations and body iron status in children

Blood lead concentrations and body iron status were investigated in 279 children. Blood lead concentrations showed no increase during iron depletion phase (stage I) but markedly increased from the phase of iron deficient erythropoiesis (stage II). Increased blood lead concentrations in anaemic subjects significantly decreased after iron supplementation.

Change in erythropoiesis with gestational age during pregnancy.

Abstract To investigate the changes in erythropoiesis during pregnancy, 342 pregnant and postpartum women were examined for reticulocyte subpopulations, reticulocyte maturity index (RMI), and serum transferrin receptor (sTfR) levels. The reticulocyte subpopulations were analyzed by flow cytometry, and the sTfR concentration was measured by an immunoenzymometric method. There were no significant differences in the values of reticulocyte subpopulations, RMI, and sTfR between first trimester and non-pregnant women. However, the RMI and sTfR concentration increased gradually from the second trimester of pregnancy, during which time values were twofold higher than those in the first trimester, and peaked in the third trimester. Then the RMI and sTfR values decreased abruptly 1–4 weeks postpartum and finally dropped to the levels of non-pregnant women 5 weeks after delivery. The maternal sTfR concentration correlated significantly with the corrected reticulocyte count (r=0.52, P<0.01), RMI (r=0.67, P<0.01), and high fluorescence reticulocytes (HFR) (r=0.62, P<0.01); however, the serum ferritin level correlated poorly with the reticulocyte subpopulations during pregnancy. In conclusion, erythropoiesis increases with gestational age during pregnancy and returns to normal 5 weeks after delivery. Measurement of sTfR in combination with RMI may improve the assessment of erythropoietic activity during pregnancy.

Serum lipid concentrations correlate more strongly with total body fat than with body mass index in obese humans.

BACKGROUND To investigate the associations of body mass index (BMI) and total body fat (TBF) vs. blood lipid concentrations, we measured six anthropometric parameters, body fat mass, and serum lipid profiles in 1529 apparently healthy adults. METHODS TBF was assessed using a body fat analyzer. Serum concentrations of triglyceride, total cholesterol, and low- or high-density lipoprotein cholesterol (LDL-C or HDL-C) were determined by standard enzymatic procedures. RESULTS Serum lipid concentrations were more strongly correlated with TBF than with BMI or waist circumference in both men and women. The mean concentrations of total cholesterol in the subjects with high fatness (TBF>95th percentile) were significantly higher than those for the subjects with low fatness (TBF<5th percentile; p<0.01), but no significant differences were observed in serum lipid levels between overweights (BMI>95th percentile) and underweights (BMI<5th percentile). The incidence of hypercholesterolemia was significantly higher in the subjects with high fatness (TBF>95th percentile) than the corresponding overweight subjects (BMI>95th percentile; p<0.01). CONCLUSION TBF is more strongly associated with serum lipid concentrations in adults, at least as compared to BMI.

Significance of fetal hemoglobin-containing erythroblasts (F blasts) and the F blast/F cell ratio in myelodysplastic syndromes

To investigate the relationship between the fetal hemoglobin-containing erythroblasts (F blasts) and apoptosis in myelodysplastic syndromes (MDS), we immunohistochemically assessed F blasts, F cells, and apoptosis in 137 patients with MDS. A marked increase in the number of F blasts in the bone marrow was identified in 116 of 137 patients (84.7%), and the number of F cells was elevated in 54 patients (39.4%). Among the erythroblasts stained by anti-glycophorin C antibody, the mean percentage of F blasts was 14.63 ± 9.17% in MDS, which was significantly higher than that in non-MDS patients with stress erythropoiesis (4.82 ± 3.35%, P < 0.01), although there were no significant differences in the number of F cells between these groups. In particular, 62 of the 137 MDS patients (45.3%) had an apparent increase in F blasts but no elevation of F cells. The apoptotic rate was significantly higher in the patients with a F blast/F cell (Fb/Fc) ratio ⩾5.0 than in those with a Fb/Fc ratio <1.0 (P < 0.01). The results indicate that F cell precursors are incapable of maturing into functioning end-stage F cells, presumably owing to apoptotic cell death. The measurement of F blasts in the bone marrow is needed for the precise evaluation of fetal-type erythropoiesis in MDS.

Bone mineral density correlates strongly with basal metabolic rate in postmenopausal women.

BACKGROUND This study investigated the relationships of bone mineral density (BMD) with body composition, basal metabolic rate (BMR), and fat distribution. METHODS We measured body mass index (BMI), anthropometrics, and BMD in 345 postmenopausal women and 224 elderly men. Total body fat (TBF), fat distribution, and BMR were assessed using a body composition analyzer. Lumbar spine and proximal femur BMDs were measured with dual-energy X-ray absorptiometry. RESULTS Lumbar spine BMD was more strongly correlated with BMR (r=0.51, p<0.01) than with lean body mass (r=0.39, p<0.01) and waist hip ratio (r=-0.28, p<0.01) in postmenopausal women. The mean values of BMR in osteoporotic women were significantly lower than those for non-osteoporotic women (p<0.01). The prevalences of osteoporosis at the sites of lumbar spine and proximal femur were 32.1% and 23.3% in the women with BMR<1230 kcal, which were significantly higher than those of osteoporosis (5.4% and 7.7%) at the corresponding sites in the women with BMR> or =1230 kcal (p<0.01). In elderly men, the incidence of osteoporosis at the proximal femur was 29.5% in the subjects with BMR<1390 kcal, significantly higher than that (2.2%) in the subjects with BMR> or =1390 kcal (p<0.01). CONCLUSION BMR is more closely associated with bone density in elderly persons, at least as compared to TBF, BMI, or lean body mass.

F-Blast Is a Useful Marker for Differentiating Hypocellular Refractory Anemia From Aplastic Anemia

Making a morphologic distinction between hypocellular refractory anemia (hypo RA) and aplastic anemia (AA) is difficult. To investigate the significance of hemoglobin F-containing erythroblasts (F-blasts) and p53 expression in making the distinction between hypo RA and acquired AA, we immunohistochemically assessed F-blasts and p53 in bone marrow specimens from 16 patients with hypo RA, 31 patients with acquired AA, and 15 hematologically normal individuals. F-blast production was elevated in 87.5% (14/16) of patients with hypo RA, but in only 3.2% (1/31) of patients with AA (P < .01). p53 was over-expressed in 75.0% (12/16) of hypo RA patients and in 6.4% (2/31) of AA patients (P < .01). The mean contents of F-blasts and p53-positive cells in patients with hypo RA were 6.31% ± 3.27% and 7.54% ± 4.36%, respectively, of all bone marrow cells, which were significantly higher than for patients with AA (0.35% ± 0.46% and 0.58% ± 1.29%, P < .01). In conclusion, a high prevalence of elevated F-blast production is noted in hypo RA, suggesting that the assessment of F-blasts in bone marrow can be used as an additional tool for differentiating hypo RA from acquired AA.

Apoptotic rate varies with detection methods in myelodysplastic syndromes: impact of method selection.

To the Editor: Although several studies have confirmed the presence of higher apoptotic rates in myelodysplastic syndromes (MDS) compared with controls, these rates varied widely from 2.5% to 56.3% in reported studies (1–5). Some questions have thus been raised as to whether apoptosis is indeed a massive process in MDS; however, the question regarding the discrepancy of the apoptotic rate in MDS is still unanswered. In this study I investigated the difference in apoptotic rates from patients with MDS using three different detection methods. Apoptosis was examined in paraffin-embedded bone marrow (BM) biopsies from 152 patients (range 46–81 yr; 83 males and 69 females) with MDS. BM samples from 20 ageand gendermatched individuals (range 42–79 yr; 12 males and eight females) with no haematologic disorders were used as controls. None of the patients had received specific therapy prior to the diagnosis. For the detection of apoptosis, a TdT-mediated dUTP nick-end labeling method (TUNEL; ApopTag kit, Oncor, MD, USA), an in situ end-labeling method (ISEL) as described by Wijsman et al. (6), and a polyclonal antibody against single-stranded DNA (ssDNA; Dako, Glostrup, Denmark) were used. An immunohistochemical assay using a streptavidin– biotin complex method, based on the binding of a polyclonal antibody to ssDNA, has been used to detect cells at an early stage of apoptosis, which preceded all detachment and DNA fragmentation. Positive controls were prepared by nicking DNA with DNAase. The proportion of apoptotic cells was defined as the apoptotic rate, which represented the percentage of apoptotic cells among 1000 bone marrow cells. Statistical differences were analyzed using the Mann–Whitney U-test. P<0.01 was considered statistically significant. There were no statistically significant differences in apoptotic rates among MDS subtypes, although refractory anemia with an excess of blasts (RAEB) showed a slightly higher apoptotic rate compared with RAEB in transformation (RAEB-t). The patients with RAEB were subdivided into two groups by the percentage of blasts: RAEB-I (defined by 5–10% blasts) and RAEB-II (defined by 11–19% blasts). No significant differences in apoptotic rates were observed between the RAEB-I and RAEB-II subgroups. The percentage of apoptotic cells was greater than 1.0% in 16.4% (25/152) and 25.7% (39/152) of patients by TUNEL and ssDNA, which was taken as a provisional cut-off value based on the mean±2SD for controls. An increase of ISELpositive cells was noted in 51.9% (79/152) of patients by more than 7.0%, the cut-off value also based on the mean±2SD for controls (Table 1). My data for positive rates by TUNEL were in accordance with the results of other investigators (2) who found that a moderate increase in apoptotic cells was observed in 15.0% of MDS patients. However, Parchardiou et al. (3) reported that more than 15% of marrow cells were ISEL-positive in 85.7% of MDS patients. Sensitivity of detection methods to apoptotic cells may have differed among the studies. It thus appears that not all cells positive for apoptosis necessarily reflect apoptotic cell death. Using the ISEL technique, Raza et al. (4) found that more than 75% of hematopoietic cells in BM biopsies showed apoptosis in half the MDS patients. Lepelley et al. (2) demonstrated that the apoptotic rate was 2.5–5% in MDS patients using the TUNEL method. In this study, apoptotic rates in MDS patients were threeor five-fold higher in ISEL than in TUNEL or ssDNA assay (P<0.01). Similarly, apoptotic rates in the control group were fouror six-fold higher in ISEL than in the TUNEL or ssDNA method (P<0.01). These results suggest that the apoptotic rate varies with detection methods and should be compared only within studies using the same techniques. In conclusion, selection of detection methods is an important factor in studying apoptosis in BM from patients Eur J Haematol 2002: 69: 62–63 Printed in UK. All rights reserved Copyright Blackwell Munksgaard 2002