Increase In MCV Research Articles

English

This study was designed to evaluate the ameliorative effect of dietary supplementation of Cnidoscolus aconitifolius leaf on anaemia and changes in erythrocyte osmotic fragility in protein energy malnourished rats. Protein energy malnutrition has been associated with anaemia and changes in osmotic fragility, deformability and lifespan of erythrocytes. In this study, protein-energy malnutrition induced in weanling male Wister rats by feeding them low protein diet for 3 weeks was associated with significantly reduced (P<0.01) haematological indices: packed cell volume (PCV), red blood cell count (RBC), white blood cell count (WBC), platelet counts and mean corpuscular count (MCV). It was also associated with increased erythrocyte osmotic fragility at 0.5- 0.7% NaCl concentration. Upon introduction of recovery diets containing 20% soya protein or 20% C. aconitifolius in place of soya protein or 10% soya proteins with 10% C. aconitifolius or commercial rat feed for 4 weeks, the recovery diet containing 10% soya and 10% C. aconitifolius caused the most significant (P<0.01) increase in platelet count, WBC, MCV, MCH and MCHC when compared with all the other treated groups and the malnourished group. The group fed with 20% C. aconitifolius in place of 20% soya protein also caused the most significant elevation in RBC, PCV and Hb compared with the malnourished group. The effects of diet containing 20% soya protein and commercial feed on PCV, Hb, platelet and all other haematological indices were not significantly different from each other (p<0.05). On osmotic fragility, the recovery diet containing a mixture of 10% C. aconitifolius and 10% soya protein produced the highest reduction of osmotic fragility, followed closely by the diet containing 20% C. aconitifolius which produced greater effects than the feed containing only soya meal as the protein source or commercial rat feed which produced the lowest reduction in osmotic fragility. From the results of this study, it can be deduced that C. aconitifolius has haematopoetic property and by reducing osmotic fragility in protein energy malnutrition, it can increase the life span of erythrocytes. Key words: Cnidoscolus aconitifolius, anaemia, osmotic fragility, protein energy malnutrition.

Prevalence of anemia is lower in post‐folic acid fortification period compared to pre‐fortification period

It is not known whether improved folate status from folic acid fortification has had any impact on the prevalence of anemia in the US. Trends in indices and prevalence of anemia and macrocytosis with a focus on comparing pre‐fortification data (1988–1994) with post‐fortification data (1999–2004) were investigated (n=26596). In men, from 1988–1994 to 2003–2004, hemoglobin and hematocrit values slightly but significantly improved from 15.1 to 15.5 g/dL (2.6%) (P&lt;.0001) and from 44.5 to 45.7% (2.6%) (P&lt;.0001), respectively. In women, the improvement was from 13.3 to 13.7 g/dL in hemoglobin (3%) (P&lt;.0001) and from 39.5 to 40.4% in hematocrit (2.3%) (P&lt;.0001). There was a modest but significant increase in MCV from 1998–1994 to 2003–2004 in men (P=0.0001). MCV increased significantly in both older (&gt;60 years) men (P=.0105) and women (P=.0141). In women but not in men, prevalence of anemia was significantly lower in 1999–2004 (10.4%) relative to 1988–1994 (7.5%, a 27.9% reduction, P=.0005). In general, prevalence of macrocytosis did not change significantly from 1988–1994 to 2003–2004. In US women, folic acid fortification may be indirectly responsible for the reduction of prevalence of anemia from 1988–1994 to 1999–2004. In the post‐fortification period, increased MCV in non‐target populations such as men and older persons may suggest an adverse effect of folic acid fortification.

Effect of Hydroxyurea in Children With Sickle Cell Disease in Saudi Arabia

The objective of this study is to demonstrate the effects of hydroxyurea in children with sickle cell disease at Madina Maternity and Children's Hospital and to evaluate its short term safety. This was a retrospective review over two years period from 2004–2006. The inclusion criteria were: 1. Children with sickle cell disease who had three or more attacks of painful crisis per year 2. Children with sickle cell disease who had two or more episodes of acute chest syndrome per year. The dose range of Hydroxyurea was 15–30 mg/kg/day. The clinical episodes and the laboratory investigations were monitored monthly. The total patients included initially were 14; 4 patients were excluded because of poor compliance to treatment. 10 patients were eligible for the study. 6 were male and 4 were female. 8 patients were Saudi and 2 patients were non Saudis. The age range was 5–15 years. The attacks of painful crisis and acute chest syndrome were significantly reduced after Hydroxyurea treatment, also laboratory investigation showed significant increase in MCV and Hemoglobin F values after Hydroxyurea. We conclude that Hydroxyurea is effective in children with sickle cell disease and had no major short term adverse effect. However long terms follow up is required to evaluate long term adverse effect.

Prevalence of hematinics deficiency amongst female students and its correction

Nutritional anemia (NA) is common in India. While iron deficiency (ID) is a well recognized cause of NA, prevalence of deficiencies of other hematinics is not systematically investigated. Seventy students of a junior class of a polytechnic and 202 inmates of girl students home were taken up for study. Students were given a questionnaire to elicit anemia related symptoms. Blood was collected for complete blood count (CBC), serum ferritin, folic acid and vitamin B12. Students of polytechnic received hematinic at bed time during their menstrual periods whereas inmates of students home received hematinic at bed time, 3 days in a week. After 6 months blood tests were repeated in those who completed the treatment. CBC was done on Coulter counter and ferritin, folic acid and vitamin B12 were assayed by chemiluminescence. Students were divided into three groups-(1) Control group with Hb 12.0 g/dl or more and ferritin 15.0 ng/ml or more; (2) ID Group with Hb 12.0 g/dl or more and ferritin less than 15.0 ng/ml; and (3) Iron Deficiency Anemia (IDA) group with Hb less tha 12.0 g/dl and ferritin less than 15.0 ng/ml. Basal parameters of three groups were compared using students t test. Change in parameters with treatment was compared using paired students t test. Median age-16 years (range 10-25). Anemia ( Hb < 12.0 g/dl)-94 (34.6%); MCV < 80 fl-153 (56.3%); MCH < 27 pg-167 (61.4%); Ferritin < 15.0 ng/ml-161 (59.2%); Folic acid < 3.5 ng/ml-34 (12.5%); Vitamin B12 < 258 pg/ml-133 (48.9%) Pre-therapy: (1) Hb, MCV, MCH and ferritin significantly lower in ID and IDA Groups compared to control group. (2) Hb, MCV, MCH and Ferritin significantly lower in IDA Group as compared to ID Group. POST-THERAPY: (1) IDA group showed significant increase in Hb, MCV, MCH, ferritin, folic acid and vitamin B12. (2) final Hb (11.26+1.07) and ferritin (7.46+4.81) in IDA Group were subnormal. (3) MCV, MCH, ferritin, folic acid and vitamin B12 increased significantly in ID Group and control group. (1) Nutritional anemia is common amongst asymptomatic young female students. (2) Deficiencies of iron, folic acid and vitamin B12 are common and coexist. (3) 105 mg elemental iron for 3 days in a week for 6 months is not adequate to correct IDA. (4) 105 mg iron for 3 days in a week is enough to correct ID. (5) Non-anemic individuals with ID have iron deficient erythropoiesis. (6) Non-anemic individuals without ID, in this cohort, also had iron deficient eryhtropoiesis.

Animal Model of Mitochondrial Dysfunction Generating Macrocytic Anemia and Myelodysplastic Bone Marrow Failure.

Studies on the etiology of MDS have been hampered by the lack of relevant animal model systems. Numerous murine models of MDS/MPD overlap syndromes have been described, but MDS/MPD overlap syndromes are now felt to be clinically distinct from MDS. Likewise, a number of murine models of myeloid leukemia have been developed in which dysplastic features arise during conversion to frank leukemia. Here we describe the hematopoietic phenotype of the PolgD257A mitochondrial mutator mouse and demonstrate that this model system fulfills the myeloid dysplasia criteria outlined in the Bethesda Proposals (Blood , 100, 238, 2002). The PolgD257A allele eliminates the proofreading activity of Polg, the sole mitochondrial DNA (mtDNA) polymerase, and PolgA/A mice rapidly accumulate mtDNA mutations. Starting at 9 months of age, PolgA/A mice display multiple features of accelerated aging and develop a progressive and ultimately lethal anemia (Science , 309, 481, 2005). In data obtained at time of necropsy for other studies, the hemoglobin of young PolgA/A mice is identical to that of controls but by 10 months of age begins to fall below 10 g/dL. Overall, while Polg+/+ mice maintain a hemoglobin of greater than 10 g/dL through 30 months of age, the hemoglobin of PolgA/A drops below 5 g/dL as the mice succumb to their anemia at 13–16 months of age. As the hemoglobin of aging PolgA/A mice falls the MCV demonstrates a striking increase, from 53.6 fL to 69.8 fL. In order to confirm these intial findings, complete blood counts have been assessed prospectively in mice of all three genotypes (Polg+/+, Polg+/A, and PolgA/A; 5 mice per genotype) every other month up to 8 months of age and monthly thereafter. Full data up to 11 months of age, as well as 12 month data from all 5 PolgA/A mice, will be presented. As expected, up to eight months of age no significant difference in hemoglobin levels are apparent, but by eight months PolgA/A mice demonstrate a significant increase in MCV (51.5 fL) compared to either Polg+/A (44.9, p=0.0015) or Polg+/+ (44.5, p=0.0010) littermate controls. The disparity in MCV between PolgA/A mice and the other genotypes is progressive, with the MCV at 9 months increasing to 53.4 fL in PolgA/A mice (N=5), compared to 44.8 (p<0.0001) in the available 9 month data from the other two genotypes (N=4). In order to determine if this macrocytic anemia is associated with defects in hematopoietic maturation, morphologic examinations were performed on the marrow of old (12–13 month old) mice of each genotype. Despite the profound anemia of PolgA/A mice, their marrow is hypercellular (95–100%) compared to littermate Polg+/+ mice (85–90%), displaying megakaryocyte dysplasia (paucity of cytoplasm and abnormal nuclear lobation) and an increase in immature MPO+ myeloid precursors. In sum, these feature fulfill the diagnostic criteria for a myeloid dysplasia without evidence of myeloproliferation. In order to determine if the macrocytic anemia with myelodysplastic bone marrow morphology seen in PolgA/A mice is the result of cell-intrinsic effects at the level of the hematopoietic stem cell, as opposed to systemic effects of mtDNA mutations, both competitive and non-competitive marrow transplants have been performed. Data at 16-weeks post-transplant will be presented.

Resolving the Riddle of Obscure Iron Deficiency Anemia.

Conventional endoscopic and radiographic methods fail to identify a source of gastrointestinal blood loss in about one third of males and post-menopausal females and in most women of reproductive age with iron deficiency anemia (IDA). Such patients are often referred for hematologic evaluation. In recent years there is increasing awareness of non-bleeding conditions with abnormal iron absorption as possible causes of unexplained IDA. Both H pylori gastritis and autoimmune atrophic gastritis are associated with impaired gastric acidity interfering with iron absorption. In a prospective study performed in a community hematology clinic, 300 consecutive IDA patients were screened for non-bleeding GI conditions including celiac disease (antiendomysial antibodies), autoimmune atrophic gastritis (hypergastrinemia with strongly positive antiparietal cell antibodies) and H pylori gastritis (IgG antibodies confirmed by urease breath test). Their mean age was 39±18 y, and 251 of 300 (84%) were women. A likely cause of IDA was identified in 93% of patients. As expected in a population of females of reproductive age, only 10% had a source of gastrointestinal bleeding identified. There were 13 new cases of adult celiac disease (4%). A history suggestive of menorrhagia was present in 32%. Autoimmune atrophic gastritis was documented in 77 IDA patients (26%) of whom 39 had coexistent H pylori infection. H pylori infection was the only positive finding in 57 patients (19%). To exclude the compounding effect of menstrual blood loss in evaluating the role of H pylori in the pathogenesis of IDA, we have focused on the 29 male IDA patients with negative gastrointestinal workup, poor initial response to oral iron treatment, and a high prevalence of H pylori infection (25 of 29). Following H pylori eradication, all patients achieved normal hemoglobin levels with follow-up periods ranging from 4 to 69 months (38±15 months mean± 1SD). This was accompanied by a significant decrease in H pylori IgG antibodies and serum gastrin. Sixteen patients discontinued iron treatment, maintaining normal hemoglobin and ferritin and may be considered cured. Remarkably, 4 of the 16 achieved normal hemoglobin without ever having received oral iron after H pylori eradication. To define the relation between IDA-associated autoimmune gastritis, and pernicious anemia, we studied 160 patients with autoimmune gastritis including 83 subjects presenting with IDA, 48 presenting with normocytic indices, and 29 with macrocytic anemia. Stratification by age cohorts of autoimmune gastritis from <20 to >60 y showed coexistent H pylori infection in 87.5% at age <20 y, 47% at 20–40 y, 37.5% at 41–60 y, but only 12.5% at age > 60y. With increasing age cohorts, there was a regular and progressive increase in MCV from 68±9 to 95±16 fl, serum gastrin from 349±247 to 800±627 u/mL, and a decrease in cobalamin from 392±179 to 108±65 pg/mL. Conclusions: The favorable long-term clinical results of H pylori eradication offer strong evidence for a cause-and-effect relation between H pylori and IDA. A proportion of H pylori patients will develop autoimmune gastritis. Our findings in autoimmune gastritis imply a disease presenting as IDA many years before the establishment of clinical cobalamin deficiency. It is likely caused by an autoimmune process triggered by antigenic mimicry between H pylori epitopes and major autoantigens of the gastric mucosa. Recognition of H pylori and autoimmune gastritis as common causes of obscure iron deficiency anemia should have a strong impact on the efficacy of diagnostic workup and management of unexplained, or refractory iron deficiency anemia.

Effect of storage on microcytosis observed in dogs with portosystemic vascular anomalies

Microcytosis is a common laboratory finding in dogs with iron deficiency and congenital portosystemic vascular anomalies (PSVA), however artefactual changes due to blood storage may occur which could mask this feature. This study evaluated the effects of storage on microcytosis in dogs with congenital PSVA. Full haematological parameters were measured on the day of sampling and following 24 h storage at room temperature, in unaffected dogs ( n = 13) and in dogs affected with PSVA ( n = 24). Storage for 24 h resulted in significantly higher MCV values in both groups of dogs ( P < 0.01). The percentage increase in MCV was greater in the control dogs (median 8.07%, range 5.64–9.31%) compared to affected dogs (median 6.05%, range 3.12–15.21%) ( P < 0.02). Storage of 1 ml EDTA blood samples at ambient temperature for 24 h prior to analysis, as occurs when samples are posted to external laboratories, will have significant effects on MCV and may mask microcytosis in dogs with PSVA.

The effects of excessive disodium ethylene diamine tetraacetic acid (Na&lt;sub&gt;2&lt;/sub&gt;EDTA) anticoagulant concentration toward hematology profile and morphology of erythrocytes in peripheral blood examination

The purpose of this study is to know whether there are differences between hematology profile and morphology of erythrocytes of blood specimens which are prepared with excessive Na2EDTA anticoagulant in different concentration. This study was conducted in Faculty of Medicine Gadjah Mada University. The criteria of subject were male, age from 18 until 22 years old and healthy, ascertained from history taking and vital sign examination. Blood samples from 33 subjects were taken using vein puncture. Two millimeters blood was divided into 4 Na2EDTA-containing tubes. Before that, one drop of blood without Na2EDTA anticoagulant was used for making control blood film right after vein puncture. Each tubes contained different concentration of anticoagulant. The first tube contained Na2EDTA in standard concentration 2 mg/dl; the remaining tubes contained consecutively, 4 mg/dl, 6 mg/dl, and 8 mg/dl. Those samples were immediately examined using SYSMEX SE-9500 automatic analyzer for measuring erythrocytes hematological profile and were stained with Wright staining for morphological examination. These procedures were done before 20 minutes of vein puncture. There were significant decrease of RBC count, HGB, HCT, and MCHC and also significant increase of MCV and RDW between different concentrations of excessive Na2EDTA anticoagulant. MCH did not have significant result. Morphological examination showed significant morphological changes in the form of echinocytes and appearance of ghost cells in the sample treated with excessive Na2EDTA anticoagulant concentration. In conclusion, there are differences in hematological profile and morphology of erythrocytes among blood specimen which are prepared with excessive Na2EDTA anticoagulant in different concentration, except for MCH. Excessive Na2EDTA anticoagulant concentration will affect the blood specimen for peripheral blood examination of erythrocytes by interfering morphology and some of hematological parameters. (Med J Indones 2006; 15:157-64) Keywords: Na2EDTA, erythrocytes, hematology parameters, echinocytes, ghost cells

The Hidden Face of Pernicious Anemia: Age-Related Progression of Autoimmune Gastritis from Iron Deficiency to Cobalamin Depletion.

Because gastric acid secretion is critical for food iron absorption, iron deficiency is a known complication of the achlorhydria associated with pernicious anemia at presentation or following cobalamin (Cbl) treatment. In a previous study employing high serum gastrin and strongly positive antiparietal cell antibodies as a screening method for autoimmune atrophic gastritis, we have found a very high (27%) prevalence of atrophic gastritis among young subjects with iron deficiency anemia (IDA). The objectives of the present study were to explore the degree of overlap between patients presenting with atrophic gastritis and microcytic (MVC<80 fl) IDA on one hand, and those presenting with Cbl deficiency and normocytic (MCV 80–100 fl) or macrocytic (MCV>100 fl) anemia typical of pernicious anemia. Of 160 patients diagnosed with autoimmune gastritis presenting with iron, Cbl, or combined deficiency over the years 2001–2005, 83 presented with microcytic IDA, 48 with normocytic and 29 with macrocytic indices. Serum Cbl was abnormal in 100% of macrocytic, 92% of normocytic and 46% of microcytic subjects. IDA patients were 21 y younger (41 ±15 vs 62±15 y), predominantly female (78 vs 41%) and with a higher proportion of active H pylori infection (42 vs 21%). However, there were also marked similarities between all subgroups including a high prevalence of thyroid disease (20%) and diabetes (8%) known for their association with the autoimmune polyendocrine syndrome, and the rate of anti-intrinsic factor positivity was the highest (42% vs 31%) among IDA patients. Stratification by age cohorts from <20 to >60 y showed a very regular correlation, with progressive increase in MCV from 68±9 to 119±8 fl, serum ferritin from 4±2 to 37±41 μg/L, hypergastrinemia from 349±247 to 800±627 u/mL (normal 61±17), and a progressive decrease of Cbl from 392±179 in the youngest, to 108±65 pg/mL in the oldest age cohort. The prevalence of H pylori infection was 87.5% at age <20 y, 47% at 20–40 y, 37.5 % at 41–60 y and 12.5% at age > 60y implying a spontaneous elimination of H pylori by achlorhydria of increasing severity and duration. H pylori eradication by triple therapy in 24 patients resulted in a decrease in serum gastrin from 476±391 to 218±220 u/mL (paired t-test=0.00086) within 19± 12 months and complete remission of pernicious anemia in 2 patients. These findings challenge the common notion that pernicious anemia is a disease of the elderly manifested in megaloblastic anemia, and imply a disease starting many years before the establishment of clinical Cbl deficiency through an autoimmune mechanism directed against gastric parietal cells, likely triggered by H pylori by means of antigenic mimicry. Because of the added strain of young age and fertility on iron requirements, IDA may precede Cbl deficiency by many years until the crucial loss of remaining intrinsic factor in a proportion of patients terminating in typical pernicious anemia.

Hydroxyurea for the Prevention of Stroke and Other Vasoocclusive Complications in Children with Sickle Cell Disease.

Due to the immigration of families of Mediterranean, Asian and African origins to Switzerland an increasing number of children with Sickle Cell Disease (SCD) are being followed in our centers. The Swiss Pediatric Sickle Cell Disease Registry is aimed to study prognostic aspects and the outcome of different therapeutic approaches. Particularly, indications for hydroxyurea treatment and the effects of its long-term use are assessed.The risk of stroke in children with sickle cell disease (SCD) is approximately 200–300-fold greater compared to healthy children. 10% of children with SCD have a specially high risk for stroke with an incidence of 1 in 100 children per year. The US stroke prevention (STOP) trial has demonstrated the benefit of regular red cell transfusions for stroke prevention; however hemochromatosis is frequent and blood-borne infections can occur. Hydroxyurea (HU) can be a therapeutic alternative for the prevention of stroke in children with SCD and other severe vasoocclusive complications (VOC).From 52 children in our centers with SCD or with compound hemoglobinopathies 27 (median age 10.5, range 3.8–19 y) are treated with HU and were followed for 9–101 (mean 47) months. 16 received HU for recurrent VOC, including 4 children with chest syndrome. Two children recieved HU for silent cerebral infarctions and after stroke, respectively (Jehovas Witness). In 9 children HU was commenced for pathological transcranial dopppler ultrasonography (TCD) results according to the STOP trial criteria. TCDs were performed in all SCD patients once a year, in children with conditional TCDs every 6 months, and every 3 months in patients with pathological TCD results. Upon observation of two pathological TCDs at 2–3 months interval, HU was initiated with 10 to maximally 30 mg/kg/day.HU was well tolerated in all patients and no leuko- or neutropenia was observed. No stroke, stroke recurrence, chest syndrome or splenic sequestration was observed during HU treatment and the average number of hospital days/y before (9d) and after 12 months of HU (3d) were reduced significantly (P=0.002). However after 12–24 months, abnormal lung function and growth retardation did not improve significantly. Laboratory studies documented a significant increase in total Hb (mean 79 to 93 g/l; P=0.03), HbF (9 to 23%; P=0.0001) and MCV (83 to 94 fl; P=0.001) and a significant decrease of HbS (86 to 71%; P<0.0001), WBC (7,8 to 5.4; P<0.05) and bilirubin (54 to 36 umol/l; P<0.05) after 12 months of HU. In children that had received HU for pathological TCDs the increase of HbF resulted in a reduction of the middle cerebral arteries blood flow velocities as soon as after 5–7 months.We conclude that HU is a promising therapeutic alternative for the prevention of stroke and other VOC in SCD. Safety and efficacy of this treatment in children has to be assessed by further long-term studies.

Effect of triploidy on turbot haematology

This study was carried out to compare key haematological features of diploid (2n) and triploid (3n) turbot as a first step towards the assessment of the ability of 3n turbot to withstand sub-optimal culture conditions. Morphometric indices of erythrocytes were determined on blood smears by light microscopy. Triploidy significantly ( P < 0.001) increased all morphometric indices measured in the erythrocytes, including size, surface, and volume, except for the size of minor nuclear axis. The increase in cell size was larger for the major (31.0%) than for the minor (8.3%) axis, thus rendering erythrocytes of 3n turbot more ellipsoidal. The increase in erythrocyte volume (45.9%) was close to the theoretical expected 50% increase as a result of one extra chromosome set. Haematological indices were measured automatically by a haematological Coulter Counter. Triploid turbot had lower numbers of red blood cells (RBC: 1.84 cells pL −1 in 2n vs. 1.27 cells pL −1 in 3n; P < 0.001) but they were of a larger size (Mean corpuscular volume [MCV]: 145.51 fL in 2n vs. 181.78 fL in 3n; P < 0.001). However, the decrease in RBC was not compensated by the increase in MCV, and thus, triploidy decreased the haematocrit (Hct: 26.80% in 2n vs. 23.11% in 3n; P < 0.001) and total blood haemoglobin concentration (Hb: 73.74 g l −1 in 2n vs. 67.54 g l −1 in 3n; P < 0.05). In contrast, mean corpuscular hemoglobin (MCH: 40.27 pg in 2n vs. 53.28 pg in 3n; P < 0.001) was higher for 3n turbot as a result of their larger erythrocytes although MCH concentration (MCHC: 0.28 pg fL −1 in 2n vs. 0.29 pg fL −1 in 3n did not significantly differ. RBC, Hct and MCV were also determined manually using light microscopy. In general, discrepancies between the two methods were small (overall ∼7%) but the Coulter Counter tended to overestimate RBC and Hct (and thus to underestimate MCV). Nevertheless, relative differences between ploidies were very similar, thus verifying triploidy-associated changes in hematological features. These changes, as determined in the present study, are essential when evaluating the feasibility of triploid turbot for intensive aquaculture systems in which unfavorable situations may occur.

Hematological and Biochemical Studies on Workers Occupationally Exposed to Pesticides

Pesticides can cause many problems in exposed individuals. Twenty-four workers exposed to pesticides have been investigated. They were divided into two groups. The first group is the applicators of pesticides [14 workers] and the second group is the salesmen who sale pesticides in the market [10 workers]. Ten non-exposed individuals were used as control. Hemoglobin content [Hb], hematocrit value [Hct], red blood cells [RBCs], white blood cells counts [WBCs], mean corpuscular volume [MCV], mean corpuscular hemoglobin [MCH], and mean corpuscular hemoglobin concentration [MCHC] were measured. Biochemical liver functions: Serum aspartate aminotransferases [AST], and alanine aminotransferases [ALT], renal functions: creatinine [CR] and uric acid [UA], acetyl cholinesterase [AChE] activity, and total protein [TP] were estimated in the occupationally and non-occupationally exposed groups. Significant decrease was observed in the hematological parameters [Hb, RBCs, WBCs], plasma AChE activity, and the total protein. In contrast, significant increase in serum AST activity, creatinine, and uric acid concentrations were record in the exposed groups when compared with the non-exposed groups. Also, significant increase in MCV and MCH values of the two exposed groups was found. Salesmen were more affected than the applicators group. These results suggested that salesmen group were exposed to various pesticides and for a long time during day by different routes of exposure.

High Susceptibility of Djungarian Hamsters (Phodopus sungorus) to the Infection with Babesia microti Supported by Hemodynamics

As the comparative study was carried out on the susceptibility by the pursuit of parasitemia among the Djungarian, Syrian, and Chinese hamsters as well as BALB/c mice infected with the Syrian hamster-adapted Babesia microti strain, and Djungarian hamsters showed the highest parasitemia among them. Then, the other hematological parameters were pursued in the Djungarian hamsters infected with the hamster-adapted B. microti strain. Remarkable symptoms observed were hemoglobinuria clinically, anemia hematologically, and splenomegaly macroscopically during all over the observation period for 24 weeks post infection (PI). Parasitemia began to rise at 2 weeks and peaked at 4 weeks PI. After that, parasitemia decreased gradually but was maintained with a level of about 10% on average until 24 weeks PI at the end of the experiment. A decrease in the RBC count, Hb, and PCV, and an increase in the reticulocyte and WBC counts due to the development of immature neutrophils, lymphocytes and monocytes were recognized together with a rise of parasitemia. The hamsters had macrocytic hypochromic anemia due to the increase of MCV and the decrease of MCHC in the growth phase of the parasite. It was considered that the Djungarian hamsters will be useful for the infection examination, isolation, maintenance, and passage of B. microti in laboratory.

Prevelence of α-Globin Deletions in β-Thalassemia Minor Patients and Differences in Erythrocyte Indices.

Introduction: β-thalassemia (thal) minor is caused by multiple β-globin (gb) gene mutations, most being located in the promotor and IVS-1. Large variations in erythrocyte indices between β-thal minor patients have been reported and are thought to be caused by the type of β-gb gene mutation. α-thal minor is caused by partial or total α-gb gene deletions. α-gb gene deletions have been reported in all racial origins. It is therefore likely that α-gb gene deletions occur in β-thal minor patients.This combination of gb genes mutations could also have an impact on erythrocyte indices.Objectives: The trial objectives were to determine the prevalence of α-gb gene deletions in β-thal minor patients and to compare erythrocyte indices between 3 groups. Group 1: no α-gb gene deletion, Group 2: deletion of 1 α-gb gene, Group 3: deletion of 2 α-gb genes.Methods: Diagnosis of β-thal minor was established using hemoglobin HPLC analysis (Variant II, Bio-Rad) with increased HbA2 +/− increased HbF without mutant Hb. The DNA of consecutive cases with newly diagnosed β-thal minor was extracted from leucocytes. A multiplex PCR assay was used to detect the presence of 7 α-globin gene deletions: −α3.7,−α4.2, −−SEA, −−FIL, −−MED, −−THAI, −α20.5. Data on age, sex and erythrocyte indices (Hb, MCV, RBC and RDW) was recorded. An ANOVA was used to compare groups. P significance was established at 0.01 to adjust for multiple analysis.Results: 300 specimen were collected in 9 months. 34 were excluded because of poor DNA quality or presence of a mutant Hb. 25 patients (9.4%) had at least 1 α-globin gene deletion. Group1 included 241 patients, Group 2: 20 patients and Group 3: 5 patients. No differences for age or sex were present between the groups. Differences in erythrocyte indices are reported in Table 1. LogMCV was used to adjust for variance heterogeneity.Conclusions: 9.4% of β-thal minor have α-gb gene deletions. The only parameter that is significantly different between groups is MCV (Hb not reaching the pre-specified level of 0.01). It is therefore possible that differences in MCV between patients with β-thal minor can be explained by α-gb gene deletions. The concommittant presence of β-gb and α-gb mutations might improve the β-gb/α-gb imbalance implicated in ineffective erythropoiesis and a significant increase in MCV (and Hb but not reaching statistical significance in this study due to the sample size). However, this parameter does not have sufficient power to differenciate or identify β-thal minor patients with α-globin gene deletions. Other factors certainly influence MVC variability in β-thal minor patients and the independant influence of this variable (α-gb gene deletions) will require further investigation.Erythrocyte indices between groupsParameterGroupMeanStd Deviation95% CIPMCV165.86573.984565.3580–66.3734<.0001270.06007.557966.5228–73.5972373.26003.601168.7886–77.7314Log MCV11.8176.026371.8142–1.8209<.000121.8432.045041.8221–1.864331.8644.021521.8377–1.8912RDW116.00041.280015.8380–16.1628.068215.45001.895814.5627–16.3373315.02002.090912.4238–17.6162RBC15.7081.73695.6144–5.8018.88525.6835.67295.3686–5.998435.5500.61814.7826–6.3174Hemoglobin1119.775914.9010117.8851–121.6668.0382127.300013.6501120.9115–133.68853129.400012.3410114.0766–144.7234

Treatment with Hydroxyurea in Sickle Cell/ ß Thalassemia: A Long-Term Experience.

Sickle cell/ß-Thalassemia is a common disease in areas where ß-Thal and ßS genes are endemic, like in Sicily. In the current study we evaluated clinical and hematological data of Sicilian patients with Sickle cell/ß-Thalassemia treated with Hydroxyurea (HU). The endpoint of the study was to evaluate the efficacy of HU in terms of reduction of sickle cell crises after 2 years of treatment in comparison with the 2 years before. Moreover, we evaluated the outcome after long-term treatment. Fortytwo patients (18 males, mean age 36, range 18–53) were treated with HU (mean dosage 15 mg/kg, range 10–30) for an average 6.6 years follow-up (range 3–9 years). Twentytwo were ß0/ßS and 20 ß+/ßS genotype. All had 3 or more crises in the year before starting HU. We observed a significant reduction in sickle cell crises (7.8 ± 6.9 crises per year versus 0.9 ± 1.8 per year, P < 0.0001), hospitalizations (2.5 ± 2.9 per year versus 0.3 ± 1.5, P < 0.0001), and days in hospital (22.4 ± 21.9 per year versus 1.2 ± 2.3, P < 0.0001). Altogether, there was a 86% reduction in vasoocclusive events in comparison with the 2 years before (P < 0.001). Moreover, there was a significant increase of MCV (71.4 versus 97.5fl, P < 0.0001), HbF (7.5 versus 25.2 %, P < 0.0001), and decrease of WBC (11.4 versus 9.2 109/L P < 0.01) and reticulocytes (14.1 versus 10.2%, P< 0.01). Finally, redution of hyperdense cells and increase of erithropoyetin were seen. After a mean follow-up of 6.6 years, 39 patients are alive. Three died (2 end stage HCV related liver disesases, 1 bleeding after ERCP). Nine of the 40 alive patients developed complications: 1 acute chest syndrome, 2 strokes, 2 myocardial infarctions, 4 bone necrosis. Brain MRI of 15 patients after and during a mean of 6.9 years of HU treatment showed 2 new onset strokes, 1 of which in a patient with a previous stroke. Moreover, 4 patients developed new onset asymptomatic ischemic brain lesions. In every case there had been a significant reduction of sickle cell crises. There were two cases of cancer, occurring in two patients who were brother and sister: lung cancer in the former, a proeviously heavy smoker, breast cancer in the latter. No further serious adverse events were seen. Five patients with iron overload were treated with Deferiprone: no drug interaction with HU was noticed. Our study confirms that HU is effective in reducing clinical relevant crises of patients with Sickle cell/ß-Thalassemia. However, our preliminary data suggest that chronic organ damages are not prevented by HU. Safety has to be assessed by more prolonged studies.

Clinico-haematological and mineral studies on experimental maduramicin toxicity in chickens

Clinico-haematological and mineral studies were carried out in experimental chickens given maduramicin medicated feed at 5 and 10 ppm for 21 days. Maduramicin medication in both medicated groups caused growth retardation. Clinical signs namely watery diarrhoea, depression, dullness and ruffled feathers were noticed in chickens from second week of the medication at 10 ppm but this effect was seen from third week in the birds given maduramicin at 5 ppm. Maduramicin medication caused significant reduction in haemoglobin in both the medicated group from day 14 and total erythrocyte count and packed cell volume in 10 ppm group on day 21. There was an increase in MCV in 10 ppm group on day 21 indicating macrocytic anaemia and decrease in mean corpuscular haemoglobin concentration (MCHC) in both the medicated groups indicating hypochromic anaemia. The leucopenia due to lymphopenia was observed in 10 ppm group on day 21. Maduramicin medication caused significant increase in serum Zn in 10 ppm group and decrease in Cu concentration in both the medicated groups from day 14. It is concluded that maduramicin caused toxic effects from day 14 in both the medicated groups.

Clinico-haematological and mineral studies on experimental maduramicin toxicity in chickens

Clinico-haematological and mineral studies were carried out in experimental chickens given maduramicin medicated feed at 5 and 10 ppm for 21 days. Maduramicin medication in both medicated groups caused growth retardation. Clinical signs namely watery diarrhoea, depression, dullness and ruffled feathers were noticed in chickens from second week of the medication at 10 ppm but this effect was seen from third week in the birds given maduramicin at 5 ppm. Maduramicin medication caused significant reduction in haemoglobin in both the medicated group from day 14 and total erythrocyte count and packed cell volume in 10 ppm group on day 21. There was an increase in MCV in 10 ppm group on day 21 indicating macrocytic anaemia and decrease in mean corpuscular haemoglobin concentration (MCHC) in both the medicated groups indicating hypochromic anaemia. The leucopenia due to lymphopenia was observed in 10 ppm group on day 21. Maduramicin medication caused significant increase in serum Zn in 10 ppm group and decrease in Cu concentration in both the medicated groups from day 14. It is concluded that maduramicin caused toxic effects from day 14 in both the medicated groups.

Relationship between red cell mean corpuscular volume and 6-thioguanine nucleotides in patients treated with azathioprine

Azathioprine (AZA) is characterized by high interindividual differences in bioavailability and metabolization. The aim of the present study was to analyze, in patients treated with AZA for various immune system disorders, whether the variation in red blood cell mean corpuscular volume (ΔMCV) could be used as an indirect estimation of the level of the active immune modifier metabolite 6-thioguanine nucleotides (6-TGN). In 43 consecutive patients treated with a stable dose of AZA for at least 6 months who were not initially anemic, the erythrocyte 6-TGN levels with routine hematologic parameters were determined two to four times at 1-month intervals. In most patients MCV significantly increased after 3 months of therapy and stabilized after 6 months. The correlation between the daily dose of AZA and the 6-TGN level was mild (r = 0.51; P <.001). A weak correlation was also found between the dose of AZA and the ΔMCV after at least 6 months of therapy (r = 0.36; P <.05). The correlation between ΔMCV and 6-TGN level, however, was much better (r = 0.74; P <.001). The lack of a significant increase in MCV after 3 to 4 months of AZA therapy reflects low 6-TGN levels, sometimes a result of undertreatment. A determination of the 6-TGN level during the first months after AZA therapy is begun will allow more accurate adaptation of the effective dose. We observed that ΔMCV could be used as an indicator of 6-TGN levels after 6 months of AZA treatment. An increase in MCV of at least 6 fL is expected to reflect a 6-TGN level of about 175 pmol/8 × 108 red blood cells (probably being within a therapeutic value). (J Lab Clin Med 2000;135:256-62)

A CASE OF MACROCYTIC ANEMIA AS A PROBABLE ADVERSE SIDE EFFECT OF 5'-DFUR

We have experienced a case of macrocytic anemia probably caused by administration of 5'-DFUR after operation for gastric cancer. A 69-year-old man who underwent a distal gastrectomy for gastic cancer in Stage IB started to receive oral administration of 5'-DFUR since about one month after the operation. On the 20th week after starting the regimen, RBC count was 172×104/mm3, Hb was 7.3g/dl, and MCV was 127.7fl, showing prominent macrocytic anemia. Blood analysis revealed that vitamin B12 and folic acid levels were over the normal levels. We thought that the anemia might be side effect of 5'-DFUR and the drug was withdrawn. After withdrawal of the drug regimen, a remission in the anemia was attained. In addition, changes in peripheral blood anlyses were studied in 20 patients administered pyrimidine antineoplasms orally. Significant increase in MCV was observed after administration compared to that before administration in them. However, those changes were chronical in all cases except ours presented here that was only one case revealing abrupt and severe anemia. Oral pyrimidine antineoplasms are often used as postoperative adjuvant chemotherapy for gastrointestinal cancers. When they are used, periodic blood analyses are mandatory. And the regimen should be withdrawn as soon as possible if macrocytic anemia is noted.

Effect of different anticoagulant, underfilling of blood sample and storage stability on selected hemogram.

We collected blood samples from 94 adult non-hematological outpatients and inpatients for complete blood count (CBC) without any flagging at Kaohsiung Medical College Hospital in order to investigate the effect of (1) different anticoagulants with Na2 EDTA vs K3 EDTA (2) the underfilling of blood collection volume (2 ml, 3. 5 ml vs standard 5 ml) (3) the difference in storage stability between 1 hour, 4 hours, 8 hours and 12 hours after venesection at room temperature on some selected hemogram parameters (WBC, RBC, hemoglobin, hematocrit, MCV, MCH, MCHC, platelet, percentage of neutrophil and lymphocyte). The automated hematology analyzer we used was SYSMEX NE-8000, (TOA, Japan). All the EDTA collection vacutainer tubes were supplied by Becton-Dickinson (New Jersey, U. S. A.) with the same lot number. Paired t- test was used for statistics. We found that values of hemoglobin, hematocrit, MCV and lymphocyte percentage collected in Na2 EDTA tubes were significantly higher than those collected in K3 EDTA (P < 0.05 for hemoglobin and lymphocyte percentage, and P all < 0.01 for others), while values of MCHC collected in Na2 EDTA were significantly lower than those collected in K3 EDTA (P < 0.05). For underfilling of blood sample, values of hematocrit and MCV with 2 ml blood volume were significantly lower than those with 5 ml blood volume (both P < 0.01), while values of MCHC with 2 ml blood volume were significantly higher than those with 5 ml blood volume (P < 0.01). When the collection blood volume was increased to 3.5 ml, there were no significant difference between values for 3.5 ml and 5ml blood volume (P all > 0.05). In the storage stability study, there was a significant sequential increase of hematocrit and MCV between 1 hour, 8 hours and 12 hours (P < 0.05 and < 0.01, respectively, for 8 hours, P all < 0.01 for 12 hours). There was also a significant sequential decrease of neutrophil percentage between 1 hour and 4, 8, 12 hours' storage at room temperature (P all < 0.01).