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The presence of erythrocyte inclusions may indicate the presence of disease. While performing a peripheral blood smear review, you observe the cells indicated by the arrows on this Wright-Giemsa-stained slide.

How would you cla In which of the following disorders would you probably observe coarse basophilic stippling on a Wright-stained peripheral blood smear? If present, how many Howell-Jolly bodies are usually seen within a given erythrocyte?

In which of the following conditions would you expect to find Howell-Jolly bodies? What abnormality is present in this slide? The descriptions refer to the appearance of th Which of the following is NOT present in normal bone marrow and would be associated with a disease or disorder? The inclusions that are indicated by the arrows on this Wright-Giemsa stained peripheral blood smear are most likely Pappenheimer bodies.

Also appropriate for medical laboratory students, and pathology residents.

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This course identifies, describes, and illustrates red blood cell inclusions as they are observed on Wright or Wright-Giemsa-stained peripheral smears.

Numerous practice and exam questions reinforce visual recognition skills. This revised course was originally part of a series of courses adapted for the web by MediaLab Inc.

Online laboratory continuing education for clinical laboratories and med techs. Sign up for LabCE's mailing list. White Blood Cell Differential Simulator. What are the effects of hemolysis on blood plasma? Intrinsic red cell defects. What are tests for HgB and purpose for them.

Lab or Diagnostic Findings: Lead Poisoning or Sideroblastic Anemia

K, NA bicarb, K ferricyancide and K cyanide reagent oxidizes Hgb to methemoglobin which combines with cyanide to form cyanomethemoglobin pigment. What are the 4 types of Microcytic anemias? What enhances Iron absorption? OJ, vitamin C, pickles, soy sauce, vinegar, alcohol.

What inhibits iron absorption? What are iron lab profile tests? How do you determine the transferring saturation of an iron level? Stage I iron deficiency. Stage II iron deficiency. Stage III iron deficiency.

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IDA in the peripheral circulation. Iron deficient anemia SX? When does an iron deficient anemia pt usually present with sx? What causes iron deficient anemia?

What is important lab result for iron deficiency vs anemia? Low reticulocyte count increased TIBC.

What would you see on a peripheral smear in iron deficiency? How do you diagnose iron overload?

Sideroblasts vs pappenheimer bodies. How do you treat HH?

  • Sideroblastic anemia or sideroachrestic anemia is a form of anemia in which the bone marrow produces ringed sideroblasts rather than healthy red blood cells ( erythrocytes). In sideroblastic anemia, the body has iron available but cannot incorporate it into hemoglobin, which red blood cells need in order to transport oxygen.
  • 15 Jan I have read in some sources that basophilic stippling is a result of sideoblastic anemia. Is this always the case? Or can one have stippling without.
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  • Although adequate iron is present, a mitochondrial defect (decreased activity of D -ALA or ferrochelatase) prevents the incorporation of iron into hemoglobin. Acquired sideroblastic anemia frequently demonstrates a "dimorphic" RBC population [microcytic and macrocytic RBCs (increased RDW)], basophilic stippling.

Thalassemia is the result of? Which alpha thalassemia is the most severe and incompatible with life?

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  1. Pappenheimer bodies. • contrast the morphologic features of Howell Jolly bodies, basophilic stippling, and Pappenheimer bodies. The nuclei may also appear oval or slightly indented. The nuclear chromatin is clumped, dense, and dark purple. Image BCI depicts an RBC with a specific inclusion called a Howell- Jolly.:
    The inclusions can be due to aggregates of iron in the cytoplasm or within mitochondria (the latter are called Pappenheimer bodies). A nucleated erythoid cell with the same inclusions (usually within mitochondria) is termed a sideroblast . Siderocytes should be distinguished from basophilic stippling (aggregates of RNA). basophilic stippling. erythrocytes display small dots at the periphery. These dots are the visualization of ribosomes. Fine stippling: normal -- reticulocyte. Coarse stippling: imagine you're in the casino surface charge on rbc is reduced as a result of increased serum proteins, particularly fibrinogen and globulins. sideroblast. Many of the company's stores have undergone major renovations or replacement since the s. Sears began to diversify in the s, adding Allstate Insurance Company in and placing Allstate representatives in its stores in Over the decades it established major national brands, such as Kenmore, Craftsman.
  2. These results suggest that red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling do not influence the automatic counting of reticulocytes. count, increased serum iron and bone marrow erythroid hyperplasia with numerous ringed sideroblasts, the diagnosis of sideroblastic anaemia was confirmed by.:
    Best Friend Gift - Personalized Christmas Gift for a Special Friend - BFF Birthday Gift - Bridesmaid or Maid of Honor Wedding Day Gift. Best Friend .. See More. basophilic stippling = ribosomes thalassemia, lead poisoning . Pappenheimer bodies = IRON granules in mitochondria = sideroblastic anemia and asplenism.
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American journal of hematology. Iron overload in mild sideroblastic anaemias" Lancet Diseases of red blood cells D50—69,74 , — Iron-deficiency anemia Plummer—Vinson syndrome Macro-: Megaloblastic anemia Pernicious anemia. Hereditary spherocytosis Minkowski—Chauffard syndrome Hereditary elliptocytosis Southeast Asian ovalocytosis Hereditary stomatocytosis.

Fanconi anemia Diamond—Blackfan anemia Acquired: Philadelphia chromosome Accelerated phase chronic myelogenous leukemia. Juvenile myelomonocytic leukemia Chronic myelomonocytic leukemia. Refractory anemia Refractory anemia with excess of blasts Chromosome 5q deletion syndrome Sideroblastic anemia Paroxysmal nocturnal hemoglobinuria Refractory cytopenia with multilineage dysplasia.

Mast cell leukemia Mast cell sarcoma Systemic mastocytosis. Diffuse cutaneous mastocytosis Erythrodermic mastocytosis Adult type of generalized eruption of cutaneous mastocytosis Urticaria pigmentosa Mast cell sarcoma Solitary mastocytoma. Acute panmyelosis with myelofibrosis Myeloid sarcoma. Myelofibrosis Acute biphenotypic leukaemia. Haemophilia A Haemophilia B X-linked sideroblastic anemia. Ornithine transcarbamylase deficiency Oculocerebrorenal syndrome Dyslipidemia: Hunter syndrome Purine—pyrimidine metabolism: Color blindness red and green, but not blue Ocular albinism 1 Norrie disease Choroideremia Other: Dyskeratosis congenita Hypohidrotic ectodermal dysplasia EDA X-linked ichthyosis X-linked endothelial corneal dystrophy.

Alport syndrome Dent's disease X-linked nephrogenic diabetes insipidus. Genetic disorder , membrane: Retrieved from " https: Myeloid neoplasia Genetic disorders by system Aplastic anemias. Infobox medical condition new. Views Read Edit View history.

In other projects Wikimedia Commons. This page was last edited on 2 February , at By using this site, you agree to the Terms of Use and Privacy Policy. A ring sideroblast visualized by Prussian blue stain.

Mastocytoma Mast cell leukemia Mast cell sarcoma Systemic mastocytosis. AML Acute panmyelosis with myelofibrosis Myeloid sarcoma. This study explored whether the number and degree of maturation of circulating reticulocytes may be altered in OSA, possibly through the effect of erythropoietin.

Fifty male adult patients with suspected OSA, normoxic during wakefulness, were studied. After nocturnal polysomnography, a blood sample was withdrawn for blood cells count, erythropoietin, iron and transferrin determination. Reticulocyte concentration and degree of immaturity [high H , medium M , or low L ] were also determined. At multiple regression, only lowest nocturnal oxygen saturation remained a significant contributor to IRF r2 0. Anticlastogenic effects of galangin against mitomycin C-induced micronuclei in reticulocytes of mice.

On the other hand, when galangin was given to mice for 7 consecutive days before MMC injection, galangin showed potent anticlastogenic effects, even at the lowest dose level of 0. Results from our in vivo studies indicate that galangin is capable of suppressing the clastogenic activity of the direct acting MMC.

Together with our earlier observations, it appears that galangin is capable of protecting cells from the toxic effects of a variety of hazardous chemicals.

Therefore, galangin may be an useful chemopreventive compound. Malnutrition and infection influence the peripheral blood reticulocyte micronuclei frequency in children. Malnutrition is a serious public health problem that affects approximately one third of all children. The relationship between malnutrition and genetic damage has been widely studied in humans and animal models. The micronucleus MN assay is useful in detecting chromosome damage induced by several factors.

The aim of this study was to evaluate the effects of infection and malnutrition on the frequency of MN in erythrocytes from the peripheral blood of well-nourished, uninfected WN and well-nourished, infected WNI children, and moderately malnourished UNM and severely malnourished UNS children, both with infection, using a flow cytometric analysis technique. The percentage of reticulocytes RETs was significantly higher 1.

In addition, the UNS group had a 2. These frequencies were significantly higher 1. The results suggest that infection and malnutrition induce DNA damage in children. Automated reticulocyte parameters for hereditary spherocytosis screening.

The laboratory diagnosis of hereditary spherocytosis HS is based on several screening and confirmatory tests; our algorithm includes clinical features, red blood cell morphology analysis and cryohaemolysis test, and, in case of positive screening, sodium dodecyl sulphate polyacrylamide gel electrophoresis as a diagnostic test. A total of samples were screened. Gel electrophoresis was applied to samples that were positive for the screening tests.

Some other 31 anaemic conditions were also studied. In conclusion, automated reticulocyte parameters might be helpful for haemolytic anaemia diagnostic orientation even for general laboratories. In combination with cryohaemolysis, they ensure an effective and time-saving screening for HS for more specialised laboratories. Giemsa-stained wet mount based method for reticulocyte quantification: The quantity of circulating reticulocytes is an important indicator of erythropoietic activity in response to a wide range of haematological pathologies.

While most modern laboratories use flow cytometry to quantify reticulocytes , most field laboratories still rely on 'subvital' staining. Here we demonstrate the utility of Giemsa's stain commonly used microbiology and parasitology in a 'subvital' manner to provide an accurate method to visualize and count reticulocytes in blood samples from normal and malaria-infected individuals. Micronucleated erythrocytes in newborn rats exposed to raltegravir placental transfer.

However, administration of raltegravir during pregnancy produces a compound which is transferred to high concentrations to the offspring. The objective of this study is to evaluate the transplacental genotoxic effect of raltegravir in newborn rats. We evaluated the number of micronucleated erythrocytes MNE , micronucleated polychromatic erythrocytes MNPCE , and polychromatic erythrocytes PCE in the peripheral blood samples of the offspring of Wistar rats treated 6 days before birth with oral administration of raltegravir.

The animals were randomly assigned to five groups as follows: In addition, the effect of these drugs on the weight and height of newborns was assessed. Genotoxicity studies are required in pregnant women to determine the risk of using raltegravir to the fetuses.

Reticulocytes are the youngest erythrocytes released from the bone marrow into the blood and they circulate for days before becoming mature erythrocytes.

In literature, there were studies about reticulocyte parameters that could help in differentiation of iron deficiency anemia IDA from vitamin B12 deficiency anemia. However, in those studies there were no data about differentiation of mixed anemia vitamin B12 deficiency and IDA. The purpose of this study is to explore a response to 'could reticulocyte parameters help in differential diagnosis of mixed anemia? The study enrolled 26 patients with IDA, 22 patients with mixed anemia, 32 patients with vitamin B12 deficiency, and 32 age and gender matched healthy controls.

In mixed anemia, MCV could be normal or decreased, and in peripheral blood smear erythrocytes cells could be morphologically normal. For this reason diagnosis of mixed anemia is not easy and needs additional laboratory investigations. So, with a simple and cheap laboratory parameter, differentiation of. Acentric, autonomously replicating extrachromosomal structures called double-minute chromosomes DMs frequently mediate oncogene amplification in human tumors.

We show that DMs can be removed from the nucleus by a novel micronucleation mechanism that is initiated by budding of the nuclear membrane during S phase. DMs containing c-myc oncogenes in a colon cancer cell line localized to and replicated at the nuclear periphery. Replication inhibitors increased micronucleation ; cell synchronization and bromodeoxyuridine—pulse labeling demonstrated de novo formation of buds and micronuclei during S phase.

The frequencies of S-phase nuclear budding and micronucleation were increased dramatically in normal human cells by inactivating p53, suggesting that an S-phase function of p53 minimizes the probability of producing the broken chromosome fragments that induce budding and micronucleation.

These data have implications for understanding the behavior of acentric DNA in interphase nuclei and for developing chemotherapeutic strategies based on this new mechanism for DM elimination. Development of a technique for quantification of reticulocytes and assessment of erythrocyte regenerative capacity in birds. To develop a reticulocyte classification scheme, optimize an avian reticulocyte staining protocol, and compare the percentages of reticulocyte types with polychromatophil percentage in blood samples from birds.

Blood samples from a red-tailed hawk and 31 ill birds. A single blood sample obtained from a red-tailed hawk Buteo jamaicensis was used to optimize the staining protocol.

For optimization of the staining protocol, 4 dilutions of whole blood with new methylene blue stain and 4 incubation times were evaluated. From samples submitted for avian CBCs, EDTA-anticoagulated whole blood samples from 31 ill birds were randomly selected and examined to compare polychromatophil and reticulocyte percentages.

Reticulocyte staining was performed in all samples by use of a 1: In Wright-Giemsa-stained blood smears, a polychromatophil percentage was similarly determined. A reticulocyte -staining protocol was optimized. Interobserver and intraobserver variations in assessment of reticulocyte and polychromatophil percentages were not significant.

Results indicated that quantification of ring-form reticulocytes provides an accurate assessment of erythrocyte regenerative capacity in birds. Flow cytometric analysis of leukocytes and reticulocytes stained with proflavine.

Proflavine, an acridine analog for industrial use, was used to stain blood cells. A drop of blood treated with ethylenediaminetetraacetic acid-2K was mixed with a 0. Leukocytes, platelets, and reticulocytes were stained but mature red blood cells were not. Chromatin in the nuclei of all leukocytes and nucleoli of lymphocytes and monocytes had greenish-yellow fluorescence, and the kind of cell could be identified by the tone and intensity of this color.

Granules in granulocytes were in green. Reticular fine-granular or granulofibrous structures in the reticulocytes were brownish. The proflavine could be used routinely in clinical laboratories because this single stain makes possible simultaneous differentiation of leukocytes and counting of reticulocytes.

During the orchestrated process leading to mature erythrocytes, reticulocytes must synthesize large amounts of hemoglobin, while eliminating numerous cellular components.

Exosomes are small secreted vesicles that play an important role in this process of specific elimination. To understand the mechanisms of proteolipidic sorting leading to their biogenesis, we have explored changes in the composition of exosomes released by reticulocytes during their differentiation, in parallel to their physical properties.

By combining proteomic and lipidomic approaches, we found dramatic alterations in the composition of the exosomes retrieved over the course of a 7-day in vitro differentiation protocol.

Our data support a previously proposed model, whereby in reticulocytes the biogenesis of exosomes involves several distinct mechanisms for the preferential recruitment of particular proteins and lipids and suggest that the respective prominence of those pathways changes over the course of the differentiation process.

Proteolipidic composition of exosomes changes during reticulocyte maturation. Reticulocyte quantification by flow cytometry, image analysis, and manual counting. Reticulocyte counting by flow cytometry with thiazole orange was compared to manual or automated counting of new methylene blue stained blood smears. Forty-nine samples were compared for manual counting from randomly chosen clinical samples. Two hundred and eighty-nine samples from bone marrow transplant patients were compared during the period before and through chemo-irradiation and engraftment.

The slopes of correlation plots were less than 1 when flow cytometric data were the dependent variable, suggesting that thiazole orange is less sensitive than new methylene blue. In a third study, samples from bone marrow transplant patients were compared after increasing the thiazole orange concentration.

The slopes from regression analysis were 0. This demonstrates that thiazole orange is more sensitive than new methylene blue because the window of analysis includes an increased fraction of mature reticulocytes. In addition, the precision of each assay as measured. Erroneously elevated immature reticulocyte counts in leukemic patients determined using a Sysmex XE hematology analyzer. The immature reticulocyte fraction IRF in peripheral blood, as determined by automated reticulocyte analysis, is calculated using the sum of medium and highly fluorescent reticulocyte numbers and provides information about erythropoietic activity in bone marrow.

The purpose of this study was to investigate erroneously elevated IRF in leukemic patients, as determined using a Sysmex XE hematology analyzer Sysmex, Kobe, Japan. Normal reticulocyte scattergram patterns show regions corresponding to reticulocytes located between matured RBCs and an upper particle UPP region, which show a continuum of non-separated fraction. The UPP represents erythroblasts and some immature reticulocytes.

As a control group, peripheral blood was taken from patients with benign hematologic diseases, and their reticulocyte scattergrams all showed a normal pattern; UPP values were all less than However, the reticulocyte scattergrams of 5 of 11 leukemia patients showed abnormal patterns and displayed a gap between RBC and reticulocyte regions. For the remaining six leukemia patients with a normal reticulocyte scattergram pattern, immature reticulocytes were not markedly increased, and UPP values were less than The findings of the present study demonstrate that IRF results may be erroneously elevated in leukemia patients and indicate that hematologists should examine reticulocyte scattergrams and UPP values carefully.

Selective modulation of membrane sphingomyelin fatty acid turnover by nigericin. A study in the rat reticulocyte. Exposure of rat reticulocytes to Nigericin produced a selective modulation of fatty acid incorporation into sphingomyelin SM of the cell membrane, via changes in SM acylation kinetics. At physiological fatty acid concentration, Nigericin accelerated 8-fold SM acylation by decreasing the apparent K m for oleate from This constitutes a novel piece of evidence for the important role of ions in SM metabolism.

Reticulocyte -derived exosomes rex are 30— nm membrane vesicles of endocytic origin released during the maturation of reticulocytes to erythrocytes upon fusion of multivesicular bodies with the plasma membrane. Proteomics analysis of rexPy confirmed their reticulocyte origin and demonstrated the presence of parasite antigens.

Our studies thus prove, for what we believe is the first time, that rex from reticulocyte -prone malarial infections are associated with splenic long-lasting memory responses. To try extrapolating these data to human infections, in vitro experiments with spleen cells of human transplantation donors were performed.

Plasma-derived exosomes from vivax malaria patients exPv were actively uptaken by human splenocytes and stimulated spleen cells leading to changes in T cell subsets. Reticulocyte -derived exosomes rex are nm membrane vesicles of endocytic origin released during the maturation of reticulocytes to erythrocytes upon fusion of multivesicular bodies with the plasma membrane. Notably, rexPy immunization of mice induced changes in PD1 - memory T cells with effector phenotype.

Further observations on the polynucleotide-induced stimulation of protein synthesis by cell-free preparations from rabbit reticulocytes. RNA stimulates the incorporation into protein of both free amino acids and of aminoacyl residues from s-RNA. Quantitatively the response of the cell-free system to RNA is similar to that of polyuridylic acid, and there appears to be competition between messenger RNA and polyuridylic acid or polyadenylic acid. Acquisition of iron from transferrin regulates reticulocyte heme synthesis.

Fe-salicylaldehyde isonicotinoylhydrazone SIH , which can donate iron to reticulocytes without transferrin as a mediator, has been utilized to test the hypothesis that the rate of iron uptake from transferrin limits the rate of heme synthesis in erythroid cells. Taken together, these results suggest the hypothesis that some step s in the pathway of iron from extracellular transferrin to intracellular protoporphyrin limits the overall rate of heme synthesis in reticulocytes.

Spurious reticulocyte profiles in dogs with large form babesiosis: Erroneously high reticulocyte counts pseudoreticulocytosis have been reported in dogs with leukemia. Pseudoreticulocytosis and an abnormal reticulocyte profile were observed in a dog with large form babesiosis presented at our institution. The aims of this retrospective study were to determine if dogs with babesiosis and other dogs had abnormal reticulocyte profiles, and to correlate these profiles with the primary diagnosis.

Cases of large form babesiosis were identified and their reticulocyte dot plots were analyzed. Dogs with abnormal reticulocyte profiles but without microscopically apparent intraerythrocytic Babesia piroplasms were identified. The reticulocyte profiles and fluorescence ratios of dogs with and without babesiosis were compared. Twenty of 92 dogs with babesiosis had abnormal reticulocyte profiles, including 8 with a separation between the reticulocyte and mature RBC plots or a continuum of reticulocytes from the RBC plot but with a higher density of dots in the middle of the "comet tail" than in the left quarter of the dot plot.

Thirteen of dogs without Babesia on the blood smear had abnormal reticulocyte profiles, including 3 with leukemia. The medium-fluorescence reticulocyte ratios tended to be higher in dogs with babesiosis and abnormal dot plots than in other dogs, whereas the high-fluorescence ratio was higher in one dog with leukemia.

Abnormal reticulocyte dot plots and atypical reticulocyte fluorescence ratios may occur in dogs with babesiosis and alert clinical pathologists to consider this diagnosis.

Micronucleated erythrocyte frequencies in old and new world primates: Nonhuman primates are of particular relevance in evaluating the potential toxicity of drugs and environmental agents. We have used previously published information and data from the present study to establish a relationship for New World NW and Old World OW primates on the basis of the frequency of spontaneous micronucleated erythrocytes MNEs observed in peripheral blood.

The results of this study indicate that NW primates have higher spontaneous MNE frequencies than OW primates, and because of this, NW primates like the common marmoset, may be suitable for evaluating the genotoxicity of chemical agents. The water channel aquaporin-1 partitions into exosomes during reticulocyte maturation: Aquaporin-1 AQP-1 , the universal water channel, is responsible for rapid response of cell volume to changes in plasma tonicity.

In the membrane of the red cell the concentration of the protein is tightly controlled. Here, we show that AQP-1 is partially lost during in vitro maturation of mouse reticulocytes and that it is associated with exosomes, released throughout this process. AQP-1 in young reticulocytes localizes to the plasma membrane and also in endosomal compartments and exosomes, formed both in vitro and in vivo. During maturation a part of the total pool of AQP-1 is differentially sorted and released via the exosomal pathway.

A proteasome inhibitor, MG, suppresses secretion of AQP-1, implying that ubiquitination is a sorting signal for its release. We further show that modulation of medium tonicity in vitro regulates the secretion of AQP-1, thus showing that extracellular osmotic conditions can drive sorting of selected proteins by the exosomal pathway. These results lead us to suggest that AQP-1 sorting into exosomes may be the mechanism by which the reticulocyte adapts to environmental changes during its maturation.

Plasmodium falciparum can invade all stages of red blood cells, while Plasmodium vivax can invade only reticulocytes. Both proteins possess a C-terminal hydrophobic transmembrane domain, which drives adhesion to reticulocytes. In this study, the complete genome information of the P.

Eleven pvrbp family genes that included 2 pseudogenes and 9 full or partial length genes were selected and used to express recombinant proteins in a wheat germ cell-free system. The expressed proteins were used to evaluate the humoral immune response with vivax malaria patients and healthy individual serum samples by protein microarray. The recombinant fragments of 9 PvRBP proteins were successfully expressed; the soluble proteins ranged in molecular weight from 16 to 34 kDa.

The functional activity of the PvRBP family on merozoite invasion remains unknown. Micronucleation in the lens epithelium following in vivo exposure to physical and chemical mutagens. Rats were exposed to cataractogenic doses of known physical and chemical genotoxic agents in order to study the efficacy of using micronuclei to monitor mutagenicity in the lens epithelium. The total numbers of micronuclei were counted in lens epithelia from rats exposed to graded doses of either kVp X-rays or the anti-leukemic drug, 1,4 dimethanesulfonoxybutane Myleran R.

The results indicate a dose-dependent incidence of micronucleation in the lens epithelium following exposure. The findings are consistent with the hypothesis that the cataractogenicity of certain agents may be related to their effect on the genome of lens epithelial cells.

Persistence and accumulation of micronucleated hepatocytes in liver of rats after repeated administration of diethylnitrosamine. A repeat-dose micronucleus assay in adult rat liver was recently developed [Mutat. This assay demonstrated a high detectability of hepatocarcinogens at relatively low doses, as indicated by dose-dependent micronucleus induction. Because the adult rat liver is known to have a long life-span, this desirable property of the assay will be an advantage in detecting micronucleated hepatocytes MNHEPs that have persisted for long periods in the liver following repeated dosing.

However, no data directly supporting the underlying mechanisms have been published to date. In the present study, we verified the mechanisms by means of pulse-labeling of micronucleated hepatocytes with the thymidine analog 5-ethynyl-2'-deoxyuridine EdU. The rodent hepatocarcinogen diethylnitrosamine DEN was repeatedly administered orally to male Crl: CD SD rats 6 weeks old for up to 2 weeks, and EdU was injected intraperitoneally on days 1, 7, or Hepatocytes were isolated by use of a non-perfusion technique at 24h, 1 week, or 2 weeks after EdU injection and analyzed for EdU incorporation and micronucleus formation.

In addition, the number of terminal deoxynucleotidyl transferase-mediated nick end-labeling TUNEL -positive cells in the liver tissue increased, suggesting selective removal of micronucleated cells.

Theoretical calculation of the cumulative MNHEP frequency on each of the days on which DEN was administered, taking into account the rate of loss, came out closer to the actual value observed in the liver micronucleus test. Taken together, these results indicate that although micronucleated cells induced in rat livers by administration of the genotoxic hepatocarcinogen DEN undergo selective removal, they. Plasmodium vivax merozoites only invade reticulocytes , a minor though heterogeneous population of red blood cell precursors that can be graded by levels of transferrin receptor CD71 expression.

The development of a protocol that allows sorting reticulocytes into defined developmental stages and a robust ex vivo P vivax invasion assay has made it possible for the first time to investigate the fine-scale invasion preference of P vivax merozoites.

Concurrent with these changes, clathrin pits disappear by 3 hours postinvasion, replaced by distinctive caveolae nanostructures. These 2 hitherto unsuspected features of P vivax invasion, a narrow preference for immature reticulocytes and a rapid remodeling of the host cell, provide important insights pertinent to the pathobiology of the P vivax infection.

Variation of micronucleated erythrocytes in peripheral blood of Sciurus aureogaster in relation to age: In some species, in which the human is included, the influence of age in the variation in the number of micronucleated erythrocytes MNE is known.

In the present work we show how the process of aging influences the number of spontaneous MNE in the gray squirrel Sciurus aureogaster. Because of the difficulty of knowing the age of each animal, 69 animals were weighed at their arrival to the laboratory and at the start of sample taking, with the supposition that the heaviest animals were the oldest and those with the lightest weight were the youngest. The major number of MNE was found in the younger animals, whereas the adults displayed less MNE P Enrichment of reticulocytes from whole blood using aqueous multiphase systems of polymers.

This paper demonstrates the enrichment of reticulocytes by centrifuging whole blood through aqueous multiphase systems AMPSs -immiscible phases of solutions of polymers that form step-gradients in density. The interfaces of an AMPS concentrate cells; this concentration facilitates the extraction of blood enriched for reticulocytes.

AMPS enrich reticulocytes from blood from both healthy and hemochromatosis donors. Varying the osmolality and density of the phases of AMPS provides different levels of enrichment and yield of reticulocytes. When used on peripheral blood from normal donors, AMPS can provide a higher yield of enriched reticulocytes and a higher proportion of reticulocytes expressing CD71 than differential centrifugation followed by centrifugation over Percoll.

Blood enriched for reticulocytes by AMPS could be useful for research on malaria. Several species of malaria parasites show a preference to invade young erythrocytes and reticulocytes ; this preference complicates in vitro cultivation of these species in human blood.

Plasmodium knowlesi malaria parasites invade normal human blood enriched for reticulocytes by AMPSs at a rate 2. Reticulocyte parameters of delta beta thalassaemia trait, beta thalassaemia trait and iron deficiency anaemia. The following reticulocyte indices were assessed: The degree of anisocytosis in reticulocytes from patients with thalassaemia is correlated with HbF. For permission to use where not already granted under a licence please go to http: Dose- response relationship of temozolomide, determined by the Pig-a, comet, and micronucleus assay.

Temozolomide TMZ , a monofunctional alkylating agent, was selected as a model compound to determine its quantitative genotoxic dose- response relationship in different tissues blood, liver, and jejunum and endpoints [Pig-a-, comet-, and micronucleus assay MNT ] in male rats. TMZ was administered p. A statistically significant increase in Pig-a mutant phenotypes was observed on day 44 starting at 7.

In addition, a statistically significant increase in cytogenetic damage, as measured by micronucleated reticulocytes , was observed starting at 3. DNA strand breaks, as detected by the comet assay, showed a dose-dependent and statistically significant increase in liver, blood, and jejunum starting at doses of 3. Overall, PoD values show a high concordance between different tissues and endpoints, underlining the suitability of this experimental design to explore quantitative dose- response relationships in a variety of different tissues and endpoints, while minimizing animal use.

The reticulocyte parameters and profile provided by the GEN'S were studied in 38 anemic patients who fulfilled the diagnostic criteria for the ACD, and the results were compared with those of 38 healthy controls in a multivariate statistical analysis using the Student t-test and the receiver operating characteristics ROC curve analysis.

This study establishes the reticulocyte parameters and reticulocyte profile as provided by the GEN'S and characteristic of patients with ACD as compared with normal subjects. This should provide the basis for further studies comparing the reticulocyte profile of patients with ACD with those found in other types of anemia.

The RNA in reticulocytes is not just debris: Reticulocytes contain both RNA and micro-organelles and represent the last stage of erythropoiesis before full maturation to red blood cells RBCs. Even though there is continuing synthesis of hemoglobin and membrane-bound proteins in reticulocytes , the small amount of RNA that they contain has been regarded as non-functional residual material.

Here we show that this residual RNA is both functional and essential for further reticulocyte maturation. Conversely, reticulocytes treated with an RNase Inhibitor were able to form normal biconcave cells. Similarly, poor survival was also seen in reticulocytes in which protein synthesis had been blocked.

To identify the signaling pathways involved we isolated RNAs in reticulocytes versus those present in fully matured erythroblasts cultured from hematopoietic stem cells. RNAs found in erythroblasts were related to exocytosis, metabolism, and signal transduction all of which are critical for maturation through reticulocyte and into a fully mature, biconcave erythrocyte.

Our results suggest that the mRNA in reticulocytes has to be translated into novel proteins that act to preserve mitochondria and maintain cell membrane integrity as reticulocytes mature. These results enhance our understanding of the final stage of erythropoiesis and may clarify why in vitro-generated reticulocytes for transfusion purposes survive poorly. Influence of in vitro hemolysis on nucleated red blood cells and reticulocyte counts. Nucleated red blood cells NRBCs and reticulocytes are early and important measures of red blood cells' RBCs turnover, but little is known on how spurious hemolysis may affect the reliability of these parameters.

Ten EDTA-anticoagulated samples were divided into three aliquots. The first was immediately tested, where-as the others defined A and B were mechanically hemolyzed by aspiration 5 and 10 times through a small-gauge needle. An increasing amount of hemolysis was produced in hemolyzed aliquots A and B.

The RBC and reticulocyte counts progressively decreased from the nonhemolyzed sample to hemolyzed aliquots 'A' and 'B'. The NRBC count increased in 3 of the 10 samples and decreased in the remaining seven. Hemolysis of venous blood samples may seriously jeopardize NRBC and reticulocyte counts. Reticulocyte hemoglobin equivalent to detect thalassemia and thalassemic hemoglobin variants. Thalassemia and iron deficiency may both result in hypochromic microcytic anemia.

Hematological algorithms that differentiate the two are mainly established in adult selected diagnostic groups. We aimed at creating an algorithm applicable in the presence of children, hemoglobin variants, and iron deficiency.

We included reticulocyte hemoglobin equivalent, ferritin, and erythrocyte count in our algorithm. Our algorithm efficiently differentiated thalassemia and thalassemic hemoglobin variants from iron deficiency in children and adults. Increased frequency of micronucleated exfoliated cells among humans exposed in vivo to mobile telephone radiations.

The health concerns have been raised following the enormous increase in the use of wireless mobile telephones throughout the world.

This investigation had been taken, with the motive to find out whether mobile phone radiations cause any in vivo effects on the frequency of micronucleated exfoliated cells in the exposed subjects.

A total of subjects including 85 regular mobile phone users exposed and 24 non-users controls had participated in this study. Exfoliated cells were obtained by swabbing the buccal-mucosa from exposed as well as sex-age-matched controls. One thousand exfoliated cells were screened from each individual for nuclear anomalies including micronuclei MN , karyolysis KL , karyorrhexis KH , broken egg BE and binucleated BN cells.

The average daily duration of exposure to mobile phone radiations is The mean of KL in controls is The value of means of KH in exposed subjects 1. Mean frequency of broken egg is found to be more in exposed subjects 0. Frequency of presence of more than one nucleus in a cell binucleated is also higher in exposed 2. Although there is a slight increase in mean frequency of KH, BE and BN in exposed subjects but the difference is not found statistically significant.

Correlation between , , and years of exposure and the frequency of MNC and TMN has been calculated and found to. Micronucleated erythrocytes in newborns of rat dams exposed to ultraviolet-A light during pregnancy; protection by ascorbic acid supplementation.

The control group was exposed to conventional light; the experimental groups were exposed to UVA nm during gestational days In some cases, ascorbic acid Asc was administered in the drinking water from gestational day 15 until delivery. Dams were sampled at h intervals during gestation, from day 16 until delivery. Analysis of the blood containing abnormal erythrocytes or giant platelets].

We have examined the influence of erythrocytes containing inclusion bodies, nucleated red cells or giant platelets on the measurement of reticulocyte count by automated machine, R Correlation of the reticulocyte count between automated and conventional method was extremely good in the blood containing red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling. However, correlation was poor when the sample contained the nucleated red cells.

Reticulocyte count was decreased in the blood with significant amounts of nucleated red cells. Since nucleated red cells themselves are not counted as reticulocytes in the machine, this was considered to be due to increased young reticulocytes which frequently appeared with nucleated red cells.

Both cold agglutinated red cells and giant platelets apparently influenced the reticulocyte count by the R These results suggest that red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling do not influence the automatic counting of reticulocytes. Although nucleated red cells, cold agglutinated red cells and giant platelets affected the reticulocyte count, the machine shows abnormal flags in most of above cases except highly agglutinated red cells , so that one can recount reticulocytes by conventional method.

We conclude the machine can safely count the reticulocytes even in the blood containing abnormal red cells or platelets. Importance of spontaneous micronucleated erythrocytes in bottlenose dolphin Tursiops truncatus to marine toxicology studies.

The objective of the work was to characterize the presence of spontaneous micronucleated erythrocytes MNES from peripheral blood of bottlenose dolphins Tursiops truncatus to evaluate the possibility to use this species as potential bioindicator of genotoxic compounds. Forty-eight blood samples from 12 bottlenose dolphins were obtain from three Mexican dolphinariums, and from 10 dolphins was possible to obtain more than one sample at different sampling times.

Smears were processed and observed with an epifluorescence microscope. No variations in the MNES values of the bottlenose dolphins that were sampled more than one occasion were found. Comparisons among dolphinariums revealed differences in MNES frequency, with the highest significant frequency observed in dolphins from dolphinarium "A" Wistar rats were exposed to 2. Four subgroups were created in order to be irradiated 4, 16, 30 and 60 hours. Sham-exposed controls were included in the study.

Animals were euthanized on the final irradiation day of each treated subgroup. Bone marrow smears were examined to determine the extent of genotoxicity after the particular treatment time. Mann-Whitney test was used for statistical evaluation of data. In comparison to the sham-exposed subgroups, the findings of polychromatic erythrocytes revealed significant differences for the 8th and 15th experimental day.

Thereafter, the frequency of micronucleated bone marrow red cells was significantly increased after 15 irradiation treatments. Comparison of micronucleus frequency data obtained after 2, 8 and 30 irradiation treatments did not reveal statistically significant differences between sham and treated subgroups. Reticulocyte hemoglobin content predicts functional iron deficiency in hemodialysis patients receiving rHuEPO.

Early detection of iron sufficiency at the level of the erythropoietic cell is necessary to optimize management of uremic anemia with recombinant human erythropoietin rHuEPO. Transferrin saturation and serum ferritin measurements have been noted to be insensitive and inaccurate measures to detect functional iron deficiency.

Recently, the reticulocyte hemoglobin content CHr has been shown to be a sensitive and specific indicator of functional iron deficiency in nondialysis patients treated with rHuEPO. In study 1, stable HD patients were studied at two outpatient dialysis centers. CHr was normally distributed, with a mean value of CHr was weakly but consistently correlated with transferrin saturation and serum ferritin. CHr and reticulocyte number were inversely correlated with red blood cell RBC number, suggesting that the erythropoietic stimulus of routinely administered rHuEPO may have resulted in functional iron deficiency.

Month-to-month changes in CHr correlated weakly with changes in serum iron and percent transferrin saturation, but not at all with changes in serum ferritin. When we analyzed those patients with baseline CHr less than 26 pg, a level strongly suggestive of functional iron deficiency, these correlations strengthened, and in addition, month-to-month changes in CHr correlated strongly and directly with concomitant changes in RBC count, hemoglobin, and hematocrit, suggesting that rising CHr was indicative of an erythropoietic response.

In study 2, 79 patients received a single-dose infusion of mg iron dextran. After intravenous iron, CHr rose within 48 hours, peaked at 96 hours, and then fell toward. VLA-4 blockade by natalizumab inhibits sickle reticulocyte and leucocyte adhesion during simulated blood flow. Titrations with natalizumab revealed the presence of saturable levels of VLA-4 on both SCD reticulocytes and leucocytes similar to healthy subject leucocytes. Under physiological flow conditions, the adhesion of SCD whole blood cells and isolated SCD leucocytes to immobilized vascular cell adhesion molecule 1 VCAM-1 was blocked by natalizumab in a dose-dependent manner, which correlated with cell surface receptor binding.

Thus, VLA-4 blockade may be beneficial in sickle cell disease. Comparison of male versus female responses in the Pig-a mutation assay. Labash, Carson; Avlasevich, Svetlana L. Validation of the Pig-a gene mutation assay has been based mainly on studies in male rodents. To determine if the mutagen-induced responses of the X-linked Pig-a gene differ in females compared to males, 7- or week old male and female Sprague Dawley rats were exposed to N-ethyl-N-nitrosourea ENU.

These results suggest that the slightly higher response in younger males than in the younger females may be related to differences in erythropoiesis function at that age.

In conclusion, while some quantitative differences were noted, there were no. Reticulocyte and erythrocyte hypochromia markers in detection of iron deficiency in adolescent female athletes. The aim of this study was to analyse the effectiveness of new haematology parameters related to reticulocytes and mature red blood cells to differentiate pre latent and latent iron deficiency.

The study included female athletes aged years representing volleyball, handball, cycling, canoeing, cross-country skiing, swimming and judo. To assess iron status the concentration of ferritin, soluble transferrin receptor sTfR , iron and total iron binding capacity TIBC were determined in serum.

In addition to blood morphology, the mean cellular haemoglobin content in erythrocytes CH and reticulocytes CHr , mean cellular haemoglobin concentration in reticulocytes CHCMr , the percentage of erythrocytes HYPOm and reticulocytes HYPOr with decreased cellular haemoglobin concentration, the percentage of erythrocytes LowCHm and reticulocytes LowCHr with decreased cellular haemoglobin content, and percentage of erythrocytes with decreased volume MICROm were determined.

Subjects with ferritin reticulocytes: CHCMr p Hexokinase microheterogeneity in rabbit red blood cells and its behaviour during reticulocytes maturation. Hexokinase in rabbit reticulocytes is present in two molecular forms hexokinase Ia and Ib separable by ion-exchange chromatography on DE columns. This new hexokinase microheterogeneity is not caused by different oxidized forms of the enzyme nor influenced by the presence of proteolytic inhibitors during lysate preparation.

Determine the yield of micronucleated cells in primary human fibroblasts exposed to focused soft X-rays. Our part of the study has been to determine the effectiveness of soft X-rays at inducing chromosomal damage under conditions of direct and bystander exposure.

Previous studies have been performed with primary human fibroblasts measuring micronuclei formation to determine the relative yields of direct versus bystander mediated micronuclei formation after cells were individually irradiated utilizing our novel focused soft X-ray microprobe, which is capable of producing localized submicron beams of carbon-K eV X-rays. Only a brief overview is given here as the study has been published in several papers.

Our original hypothesis was to study yields of bystander-induced micronucleated cells in both wild-type and mutant fibroblast from mouse embryo fibroblasts.

Difficulties with the level of background micronuclei in the MEFs prevented systematic studies of bystander responses in the laboratories involved in the collaboration. Our soft X-ray source has been in routine use for carbon-K X-rays and is now available with Aluminium-K 1. A separately funded project developed a new soft X-ray microprobe which will give much greater flexibility for changing energies and giving high dose. In vivo erythrocyte micronucleus assay III.

Validation and regulatory acceptance of automated scoring and the use of rat peripheral blood reticulocytes , with discussion of non-hematopoietic target cells and a single dose-level limit test. The in vivo micronucleus assay working group of the International Workshop on Genotoxicity Testing IWGT discussed new aspects in the in vivo micronucleus MN test, including the regulatory acceptance of data derived from automated scoring, especially with regard to the use of flow cytometry, the suitability of rat peripheral blood reticulocytes to serve as the principal cell population for analysis, the establishment of in vivo MN assays in tissues other than bone marrow and blood for example liver, skin, colon, germ cells , and the biological relevance of the single-dose-level test.

Our group members agreed that flow cytometric systems to detect induction of micronucleated immature erythrocytes have advantages based on the presented data, e. Flow cytometric analysis of peripheral blood reticulocytes has the potential to allow monitoring of chromosome damage in rodents and also other species as part of routine toxicology studies. It appears that it will be applicable to humans as well, although in this case the possible confounding effects of splenic activity will need to be considered closely.

Also, the consensus of the group was that any system that meets the validation criteria recommended by the IWGT should be acceptable. A number of different flow cytometric-based micronucleus assays have been developed, but at the present time the validation data are most extensive for the flow cytometric method using anti-CD71 fluorescent staining especially in terms of inter-laboratory collaborative data.

Whichever method is chosen, it is desirable that each laboratory should determine the minimum sample size required to ensure that scoring error is maintained below the level of animal-to-animal variation.

In the second IWGT, the potential to use rat peripheral blood reticulocytes as target cells for the micronucleus assay was discussed. This study assessed the effect of lead Pb exposure on the status of reticulocyte count indices in workers occupied in lead battery plants. The present cross-sectional study was carried out on male lead battery workers.

The association between BLL and reticulocyte count indices was positive and significant. Genotoxic exposure to chemical substances is common, and nursing mothers could transmit harmful substances or their metabolites to their offspring through breast milk. We explored the possibility of determining genotoxic effects in the erythrocytes of breastfeeding rat pups whose mothers received a genotoxic compound while nursing.

Ten groups of female rats and five pups per dam were studied. The control group received sterile water, and the experimental groups received one of three different doses of cyclophosphamide, colchicine, or cytosine-arabinoside. There were increased numbers of micronucleated erythrocytes MNEs and micronucleated polychromatic erythrocytes MNPCEs in the samples from pups in the experimental groups P Micronucleated erythrocytes in newborns rats exposed to three different types of ultraviolet-A UVA lamps from commonly uses devices.

Exposure to ultraviolet-A UVA light can accidentally cause adverse effects in the skin and eyes. UVA radiation is emitted by lamps from everyday devices. In adult rats, micronucleated erythrocytes MNE are removed from the circulation by the spleen. However, in newborn rats, MNE have been observed in peripheral blood erythrocytes. The objective of this study was to use micronucleus tests to evaluate the DNA damage caused in newborn rats exposed to UVA light from three different types of UVA lamps obtained from commonly used devices: Rat neonates were exposed to UVA lamps for 20min daily for 6days.

The neonates were sampled every third day, and the numbers of MNE and micronucleated polychromatic erythrocytes MNPCE in the peripheral blood were determined. Many hematology laboratories have adopted semi-automated digital platforms for routine use and the evidence supporting their use is increasing. The CellaVision platforms are among the most thoroughly studied digital hematology platforms; we wished to determine the accuracy of CellaVision for reticulocyte counting.

Design, Materials and Methods: We analyzed the results for pair-wise correlation and bias, and precision. The CellaVision reticulocyte counts correlated most strongly with those of the analyzer often considered the benchmark test ; the reticulocyte count distributions were noted not to be significantly different from each other across all three methods.

The mean and standard deviation of bias were lowest in the comparison of CellaVision and LH counts. Our data provide additional support for the accuracy of digital hematology applications using the CellaVision DM96 platform. Erythrocyte antigen and reticulocyte engraftment after allogeneic hematopoietic stem cell transplantation. The aim of this study was to study the usefulness of erythrocyte antigen EA measurement to study engraftment after allogeneic HSCT.

Apart from the ABO group, 15 EAs representing six minor blood groups were followed by the simple tube agglutination technique. A total of 20 The earliest donor type EA detected was from the Rh and Kidd system. These data suggest that EA and reticulocyte assays are useful in monitoring engraftment. People donating blood more than twice annually are at risk of developing iron deficiency. Little is known about the iron status of dogs enrolled in blood donor programs.

Mature red blood cell RBC indices, reticulocyte indices, serum iron, serum ferritin, and total iron-binding capacity TIBC were compared between groups. Bone marrow is the dose-limiting organ in targeted radionuclide therapy. Hence, determination of the absorbed dose to bone marrow from incorporated radionuclides is a critical element in treatment planning. This study investigated the potential of the micronucleus assay in peripheral blood reticulocytes MnRETs as an in vivo biologic dosimeter for bone marrow.

After intravenous administration of 32P-orthophosphate or 90Y-citrate in Swiss Webster mice, DNA damage induced in bone marrow erythroblastoid cells was measured by subsequent scoring of MnRETs in peripheral blood. The response to exponentially decreasing dose rates was calibrated by irradiating animals with external Cs-gamma-rays. The maximum MnRETs frequency occurred on the second and third day after injection of 90Y-citrate and 32P-orthophosphate, respectively.

The same pattern was observed for exponentially decreasing dose rates of Cs-gamma-rays. For each type of exposure, the maximum MnRETs frequency increased in a dose-dependent manner. Using the calibrated dosimeter, the initial dose rates to the marrow per unit of injected activity were 0. Micronuclei in peripheral blood reticulocytes can be used as a noninvasive biologic dosimeter for measuring absorbed dose rate and absorbed dose to bone marrow from incorporated radionuclides.

Age, beta thalassaemia trait, and iron-deficient anaemia significantly affect reticulocyte indices in pre-school children. Reticulocyte indices are easy to obtain, low cost parameters and have gained interest in the field of diagnosing anaemias of childhood.

In the beta-thal group, HbA 2 is strongly related to all reticulocyte indices. CHr achieved the best AUC 0. Age, IDA and beta-thal significantly affect reticulocyte indices.

Using various reticulocyte fractions reticulocyte protease, inhibitor-free protease, ubiquitin and inhibitor in the presence or absence of ATP, we found that the repression of an endogenous inhibitor, as suggested by others for alpha-casein proteolysis, is unlikely for bovine serum albumin.

Therefore, differences exist in the ATP-dependent proteolytic pathway of rabbit reticulocytes depending on the substrate. Fractionation of the reticulocyte ATP-dependent proteolytic system revealed at least two proteolytic and two inhibitory fractions involved in the proteolysis of bovine serum albumin. Automated analysis of mature red blood cells and reticulocytes in SS and SC disease.

Phenotypic expression of sickle cell disease SCD is highly variable.

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Greek schistos for "divided" or schistein for "to split" injury from damaged endothelium and is a characteristic feature of microangiopathic disease of blood vessels hemolytic anemia.

AKA target cell hemoglobin E and beta thalassemia. Associated with myelophthisic anemia, commonly caused by myelofibrosis. Below 'c usually 'c for 24 months contains coagulation factors. Cryoprecipitate thawed from FFP. G6PD, PK deficiency spherocytosis sickle cell anemia.

Iron def late thalassemia Lead poisoning sideroblastic anemia - dysfunctional heme synthesis. Extensive Heinz body formation and attachment to the membrane increase the rigidity of the red blood cells and render them susceptible to fragmentation and entrapment in the spleen where they are phagocytized.

Diagnosis of an oxidant-induced hemolytic anemia is based on finding a regenerative anemia with characteristic red blood cell morphologic abnormalities of Heinz bodies and eccentrocytes. Some causes of oxidant injury also cause intravascular red blood cell lyse rupturing of red blood cell membranes and manifest with hemoglobinemia and hemoglobinuria.

Note, that methemoglobemia, Heinz bodies or eccentrocytes may dominate in a particular condition. Causes of oxidant-induced hemolytic anemia are:. Usually, but not always, the host erythrocyte contains only a single Howell-Jolly body, which appears as a blue, perfectly round inclusion.

Occasionally, uneven staining of an Howell-Jolly body can result in a ring-like appearance which may mimic a parasite. In dogs and ruminants, Howell-Jolly bodies are not routinely observed in peripheral blood because they are removed from red blood cells by the pitting function of the sinusoidal spleen.

Siderocytes are anucleate erythrocytes with iron-containing siderotic cytoplasmic inclusions. The relative simplicity of the micronucleated erythrocyte endpoint has made it amenable to automated scoring approaches. Flow cytometry is one such scoring platform that has been employed successfully. This review describes the evolution and properties of flow cytometry-based scoring of micronucleated erythrocytes.

The methodology has become widely applied to rodent blood specimens and the high throughput nature of the technology provides a number of advantages over manual microscopic scoring. For instance, the ability to efficiently survey many dose levels and many more cells per specimen relative to microscopy benefits studies that are designed to identify no observable effect levels or lowest observable effect levels.

Furthermore, flow cytometry makes it practical to study species with low spontaneous reticulocyte RET counts and micronucleus MN frequencies, thereby facilitating integration of blood-based micronucleated reticulocyte MN-RET frequency measurements into experiments conducted across species of toxicological interest.

This capability enhances genotoxicity assessments that have historically been made in dedicated MN tests performed in one species. Importantly, the feasibility of using MN-RET frequencies in blood from humans as an index of genetic damage in bone marrow opens a critical area of application that had not been practical previously. We conclude with recommendations for additional work that is needed to more fully realise the potential of flow cytometric in vivo MN scoring. Glucose permeability in naturally occurring reticulocytes and red cells from newborn piglets.

The loss of facilitated glucose transport of red cells occurring in the newborn pig was monitored in 11 density-separated cells from birth to a 4 wk of age. At birth there was a threefold increase in glucose permeability from the lightest cells to the most dense, suggesting that cells having progressively less glucose permeability are released into the circulation as gestation proceeds.

Because of extraordinary stimulation of erythropoietic activity, the uppermost top fraction constituting percent of the total cells is composed purely of reticulocytes in the growing animal. The glucose permeability of these reticulocytes which at birth has a slow but significant rate of 3. Moreover, reticulocytes themselves discard their membrane permeability to glucose in the course of maturation to red cells.

Thus, even though reticulocytes at birth are permeable to glucose, they will become red cells practically impervious to glucose within a few days. These findings suggest that the transition from a glucose- permeable fetal state to a glucose-impermeable postnatal state is brought about by two mechanisms: Azidothymidine AZT is a nucleoside reverse transcriptase inhibitor NRTI that is used for reducing mother-to-child transmission of human immunodeficiency virus I.

Since p53 plays a key role in human and mouse tumorigenesis, phaplodeficient mice are currently being evaluated as a model for assessing the carcinogenicity of perinatal exposure to NRTIs. Manual and automated reticulocyte counts. Manual reticulocyte counts were examined under light microscopy, using the property whereby supravital stain precipitates residual ribosomal RNA versus the automated flow methods, with the suggestion that in the latter there is greater precision and an ability to determine both mature and immature reticulocyte fractions.

This work aimed to compare manual and automated methodologies for reticulocyte countings and evaluate random and systematic errors. Thus, it has been confirmed that both methods, when well conducted, can reflect precisely the reticulocyte counts for adequate clinical use.

Normal and disordered reticulocyte maturation. Reticulocyte remodeling has emerged as an important model for the understanding of vesicular trafficking and selective autophagy in mammalian cells. This review covers recent advances in our understanding of these processes in reticulocytes and the role of these processes in erythroid development. Enucleation is caused by the coalescence of vesicles at the nuclear-cytoplasmic junction and microfilament contraction. Mitochondrial elimination is achieved through selective autophagy, in which mitochondria are targeted to autophagosomes, and undergo subsequent degradation and exocytosis.

The mechanism involves an integral mitochondrial outer membrane protein and general autophagy pathways. Plasma membrane remodeling, and the elimination of certain intracellular organelles occur through the exosomal pathway. Vesicular trafficking and selective autophagy have emerged as central processes in cellular remodeling.

In reticulocytes , this includes enucleation and the elimination of all membrane-bound organelles and ribosomes. Ubiquitin-like conjugation pathways, which are required for autophagy in yeast, are not essential for mitochondrial clearance in reticulocytes. Thus, in higher eukaryotes, there appears to be redundancy between these pathways and other processes, such as vesicular nucleation.

Future studies will address the relationship between autophagy and vesicular trafficking, and the significance of both for cellular remodeling. Sensitivity of reticulocyte indices to iron therapy in an intensely training athlete. Iron deficiency anaemia, and its debilitating effect on performance, is an area of concern for many female athletes.

Automated technologies that analyse individual reticulocytes may provide a sensitive measure of bone marrow response to iron supplementation.

The reticulocyte characteristics of a female volleyball player with frank iron deficiency anaemia, and her subsequent response to oral iron therapy, are reported.

Reticulocytes are immature erythrocytes; the number of reticulocytes in the peripheral blood reflects erythropoietic activity. Two cases are described to illustrate the use of the reticulocyte count in the diagnostic workup of anaemia. The first patient was a year-old woman presenting with fatigue. Laboratory evaluation showed severe macrocytic anaemia, thrombocytopaenia and the presence of schistocytes.

A low reticulocyte count suggested decreased erythropoiesis underlying the anaemia; this led to the diagnosis of vitamin B12 deficiency. The second patient, a year-old woman, also presented with fatigue and macrocytic anaemia.

A high reticulocyte count indicated increased erythrocyte degradation, and the patient was eventually diagnosed with autoimmune haemolytic anaemia. The role of reticulocytes in the differential diagnostic workup of anaemia was explored on the basis of these case descriptions. The test methodology, analytical performance, reference values and pitfalls were discussed, as well as the reticulocyte indices and their use in monitoring therapy. Reticulocytes in sports medicine: Reticulocytes are young red blood cells which develop from erythroblasts and circulate in the bloodstream for about days before maturing into erythrocytes.

With the introduction of reticulocyte count in equations and statistical models for detecting suspected blood doping, its application to sports medicine has attracted growing interest in reticulocyte behavior during training and competition seasons in athletes and experimental blood doping treatment in healthy volunteers.

An update on recent publications is therefore needed to improve the interpretation of reticulocyte analysis and its variability in sportsmen. Reticulocyte count constitutes a robust parameter during the preanalytical phase, but cell stability can be assured only if blood samples are kept at constantly cold temperatures 4 degrees C and test results will differ depending on the blood analyzer system used.

Marked intraindividual variability is the principal finding to be evaluated when exercise-induced changes are observed or illicit procedures suspected. Furthermore, reticulocyte variability is greater than that of other hematological parameters such as hemoglobin or hematocrit.

Ideally, any variation should be interpreted against long-term time series for the individual athlete: Reticulocyte distribution in athletes is similar to that found in the general population, and a gender effect in some sports disciplines or selected athlete groups may be seen. Reticulocyte variability is strongly influenced by seasonal factors linked to training and competition schedules and by the type of sports discipline. Published experimental data have confirmed the high sensitivity of reticulocyte analysis in identifying abnormal bone marrow stimulation by either erythropoietin administration or blood withdrawal and reinfusion.

Reticulocytes in untreated obstructive sleep apnoea. The short, repetitive hypoxaemic episodes observed in obstructive sleep apnoea OSA may determine small augmentations in mature red blood cells. It is unknown whether they affect reticulocyte release. This study explored whether the number and degree of maturation of circulating reticulocytes may be altered in OSA, possibly through the effect of erythropoietin.

Fifty male adult patients with suspected OSA, normoxic during wakefulness, were studied. After nocturnal polysomnography, a blood sample was withdrawn for blood cells count, erythropoietin, iron and transferrin determination.

Reticulocyte concentration and degree of immaturity [high H , medium M , or low L ] were also determined. At multiple regression, only lowest nocturnal oxygen saturation remained a significant contributor to IRF r2 0. Anticlastogenic effects of galangin against mitomycin C-induced micronuclei in reticulocytes of mice. On the other hand, when galangin was given to mice for 7 consecutive days before MMC injection, galangin showed potent anticlastogenic effects, even at the lowest dose level of 0.

Results from our in vivo studies indicate that galangin is capable of suppressing the clastogenic activity of the direct acting MMC.

Together with our earlier observations, it appears that galangin is capable of protecting cells from the toxic effects of a variety of hazardous chemicals.

Therefore, galangin may be an useful chemopreventive compound. Malnutrition and infection influence the peripheral blood reticulocyte micronuclei frequency in children. Malnutrition is a serious public health problem that affects approximately one third of all children.

The relationship between malnutrition and genetic damage has been widely studied in humans and animal models. The micronucleus MN assay is useful in detecting chromosome damage induced by several factors. The aim of this study was to evaluate the effects of infection and malnutrition on the frequency of MN in erythrocytes from the peripheral blood of well-nourished, uninfected WN and well-nourished, infected WNI children, and moderately malnourished UNM and severely malnourished UNS children, both with infection, using a flow cytometric analysis technique.

The percentage of reticulocytes RETs was significantly higher 1. In addition, the UNS group had a 2. These frequencies were significantly higher 1. The results suggest that infection and malnutrition induce DNA damage in children. Automated reticulocyte parameters for hereditary spherocytosis screening. The laboratory diagnosis of hereditary spherocytosis HS is based on several screening and confirmatory tests; our algorithm includes clinical features, red blood cell morphology analysis and cryohaemolysis test, and, in case of positive screening, sodium dodecyl sulphate polyacrylamide gel electrophoresis as a diagnostic test.

A total of samples were screened. Gel electrophoresis was applied to samples that were positive for the screening tests. Some other 31 anaemic conditions were also studied. In conclusion, automated reticulocyte parameters might be helpful for haemolytic anaemia diagnostic orientation even for general laboratories.

In combination with cryohaemolysis, they ensure an effective and time-saving screening for HS for more specialised laboratories. Giemsa-stained wet mount based method for reticulocyte quantification: The quantity of circulating reticulocytes is an important indicator of erythropoietic activity in response to a wide range of haematological pathologies.

While most modern laboratories use flow cytometry to quantify reticulocytes , most field laboratories still rely on 'subvital' staining. Here we demonstrate the utility of Giemsa's stain commonly used microbiology and parasitology in a 'subvital' manner to provide an accurate method to visualize and count reticulocytes in blood samples from normal and malaria-infected individuals. Micronucleated erythrocytes in newborn rats exposed to raltegravir placental transfer.

However, administration of raltegravir during pregnancy produces a compound which is transferred to high concentrations to the offspring. The objective of this study is to evaluate the transplacental genotoxic effect of raltegravir in newborn rats. We evaluated the number of micronucleated erythrocytes MNE , micronucleated polychromatic erythrocytes MNPCE , and polychromatic erythrocytes PCE in the peripheral blood samples of the offspring of Wistar rats treated 6 days before birth with oral administration of raltegravir.

The animals were randomly assigned to five groups as follows: In addition, the effect of these drugs on the weight and height of newborns was assessed. Genotoxicity studies are required in pregnant women to determine the risk of using raltegravir to the fetuses. Reticulocytes are the youngest erythrocytes released from the bone marrow into the blood and they circulate for days before becoming mature erythrocytes. In literature, there were studies about reticulocyte parameters that could help in differentiation of iron deficiency anemia IDA from vitamin B12 deficiency anemia.

However, in those studies there were no data about differentiation of mixed anemia vitamin B12 deficiency and IDA. The purpose of this study is to explore a response to 'could reticulocyte parameters help in differential diagnosis of mixed anemia?

The study enrolled 26 patients with IDA, 22 patients with mixed anemia, 32 patients with vitamin B12 deficiency, and 32 age and gender matched healthy controls. In mixed anemia, MCV could be normal or decreased, and in peripheral blood smear erythrocytes cells could be morphologically normal.

For this reason diagnosis of mixed anemia is not easy and needs additional laboratory investigations. So, with a simple and cheap laboratory parameter, differentiation of. Acentric, autonomously replicating extrachromosomal structures called double-minute chromosomes DMs frequently mediate oncogene amplification in human tumors. We show that DMs can be removed from the nucleus by a novel micronucleation mechanism that is initiated by budding of the nuclear membrane during S phase.

DMs containing c-myc oncogenes in a colon cancer cell line localized to and replicated at the nuclear periphery. Replication inhibitors increased micronucleation ; cell synchronization and bromodeoxyuridine—pulse labeling demonstrated de novo formation of buds and micronuclei during S phase.

The frequencies of S-phase nuclear budding and micronucleation were increased dramatically in normal human cells by inactivating p53, suggesting that an S-phase function of p53 minimizes the probability of producing the broken chromosome fragments that induce budding and micronucleation.

These data have implications for understanding the behavior of acentric DNA in interphase nuclei and for developing chemotherapeutic strategies based on this new mechanism for DM elimination.

Development of a technique for quantification of reticulocytes and assessment of erythrocyte regenerative capacity in birds. To develop a reticulocyte classification scheme, optimize an avian reticulocyte staining protocol, and compare the percentages of reticulocyte types with polychromatophil percentage in blood samples from birds.

Blood samples from a red-tailed hawk and 31 ill birds. A single blood sample obtained from a red-tailed hawk Buteo jamaicensis was used to optimize the staining protocol. For optimization of the staining protocol, 4 dilutions of whole blood with new methylene blue stain and 4 incubation times were evaluated. From samples submitted for avian CBCs, EDTA-anticoagulated whole blood samples from 31 ill birds were randomly selected and examined to compare polychromatophil and reticulocyte percentages.

Reticulocyte staining was performed in all samples by use of a 1: In Wright-Giemsa-stained blood smears, a polychromatophil percentage was similarly determined. A reticulocyte -staining protocol was optimized. Interobserver and intraobserver variations in assessment of reticulocyte and polychromatophil percentages were not significant. Results indicated that quantification of ring-form reticulocytes provides an accurate assessment of erythrocyte regenerative capacity in birds.

Flow cytometric analysis of leukocytes and reticulocytes stained with proflavine. Proflavine, an acridine analog for industrial use, was used to stain blood cells. A drop of blood treated with ethylenediaminetetraacetic acid-2K was mixed with a 0. Leukocytes, platelets, and reticulocytes were stained but mature red blood cells were not. Chromatin in the nuclei of all leukocytes and nucleoli of lymphocytes and monocytes had greenish-yellow fluorescence, and the kind of cell could be identified by the tone and intensity of this color.

Granules in granulocytes were in green. Reticular fine-granular or granulofibrous structures in the reticulocytes were brownish. The proflavine could be used routinely in clinical laboratories because this single stain makes possible simultaneous differentiation of leukocytes and counting of reticulocytes.

During the orchestrated process leading to mature erythrocytes, reticulocytes must synthesize large amounts of hemoglobin, while eliminating numerous cellular components. Exosomes are small secreted vesicles that play an important role in this process of specific elimination. To understand the mechanisms of proteolipidic sorting leading to their biogenesis, we have explored changes in the composition of exosomes released by reticulocytes during their differentiation, in parallel to their physical properties.

By combining proteomic and lipidomic approaches, we found dramatic alterations in the composition of the exosomes retrieved over the course of a 7-day in vitro differentiation protocol. Our data support a previously proposed model, whereby in reticulocytes the biogenesis of exosomes involves several distinct mechanisms for the preferential recruitment of particular proteins and lipids and suggest that the respective prominence of those pathways changes over the course of the differentiation process.

Proteolipidic composition of exosomes changes during reticulocyte maturation. Reticulocyte quantification by flow cytometry, image analysis, and manual counting. Reticulocyte counting by flow cytometry with thiazole orange was compared to manual or automated counting of new methylene blue stained blood smears. Forty-nine samples were compared for manual counting from randomly chosen clinical samples.

Two hundred and eighty-nine samples from bone marrow transplant patients were compared during the period before and through chemo-irradiation and engraftment. The slopes of correlation plots were less than 1 when flow cytometric data were the dependent variable, suggesting that thiazole orange is less sensitive than new methylene blue.

In a third study, samples from bone marrow transplant patients were compared after increasing the thiazole orange concentration. The slopes from regression analysis were 0. This demonstrates that thiazole orange is more sensitive than new methylene blue because the window of analysis includes an increased fraction of mature reticulocytes.

In addition, the precision of each assay as measured. Erroneously elevated immature reticulocyte counts in leukemic patients determined using a Sysmex XE hematology analyzer. The immature reticulocyte fraction IRF in peripheral blood, as determined by automated reticulocyte analysis, is calculated using the sum of medium and highly fluorescent reticulocyte numbers and provides information about erythropoietic activity in bone marrow.

The purpose of this study was to investigate erroneously elevated IRF in leukemic patients, as determined using a Sysmex XE hematology analyzer Sysmex, Kobe, Japan. Normal reticulocyte scattergram patterns show regions corresponding to reticulocytes located between matured RBCs and an upper particle UPP region, which show a continuum of non-separated fraction.

The UPP represents erythroblasts and some immature reticulocytes. As a control group, peripheral blood was taken from patients with benign hematologic diseases, and their reticulocyte scattergrams all showed a normal pattern; UPP values were all less than However, the reticulocyte scattergrams of 5 of 11 leukemia patients showed abnormal patterns and displayed a gap between RBC and reticulocyte regions.

For the remaining six leukemia patients with a normal reticulocyte scattergram pattern, immature reticulocytes were not markedly increased, and UPP values were less than The findings of the present study demonstrate that IRF results may be erroneously elevated in leukemia patients and indicate that hematologists should examine reticulocyte scattergrams and UPP values carefully. Selective modulation of membrane sphingomyelin fatty acid turnover by nigericin. A study in the rat reticulocyte.

Exposure of rat reticulocytes to Nigericin produced a selective modulation of fatty acid incorporation into sphingomyelin SM of the cell membrane, via changes in SM acylation kinetics. At physiological fatty acid concentration, Nigericin accelerated 8-fold SM acylation by decreasing the apparent K m for oleate from This constitutes a novel piece of evidence for the important role of ions in SM metabolism. Reticulocyte -derived exosomes rex are 30— nm membrane vesicles of endocytic origin released during the maturation of reticulocytes to erythrocytes upon fusion of multivesicular bodies with the plasma membrane.

Proteomics analysis of rexPy confirmed their reticulocyte origin and demonstrated the presence of parasite antigens. Our studies thus prove, for what we believe is the first time, that rex from reticulocyte -prone malarial infections are associated with splenic long-lasting memory responses.

To try extrapolating these data to human infections, in vitro experiments with spleen cells of human transplantation donors were performed. Plasma-derived exosomes from vivax malaria patients exPv were actively uptaken by human splenocytes and stimulated spleen cells leading to changes in T cell subsets.

Reticulocyte -derived exosomes rex are nm membrane vesicles of endocytic origin released during the maturation of reticulocytes to erythrocytes upon fusion of multivesicular bodies with the plasma membrane.

Notably, rexPy immunization of mice induced changes in PD1 - memory T cells with effector phenotype. Further observations on the polynucleotide-induced stimulation of protein synthesis by cell-free preparations from rabbit reticulocytes.

RNA stimulates the incorporation into protein of both free amino acids and of aminoacyl residues from s-RNA. Quantitatively the response of the cell-free system to RNA is similar to that of polyuridylic acid, and there appears to be competition between messenger RNA and polyuridylic acid or polyadenylic acid.

Acquisition of iron from transferrin regulates reticulocyte heme synthesis. Fe-salicylaldehyde isonicotinoylhydrazone SIH , which can donate iron to reticulocytes without transferrin as a mediator, has been utilized to test the hypothesis that the rate of iron uptake from transferrin limits the rate of heme synthesis in erythroid cells.

Taken together, these results suggest the hypothesis that some step s in the pathway of iron from extracellular transferrin to intracellular protoporphyrin limits the overall rate of heme synthesis in reticulocytes. Spurious reticulocyte profiles in dogs with large form babesiosis: Erroneously high reticulocyte counts pseudoreticulocytosis have been reported in dogs with leukemia. Pseudoreticulocytosis and an abnormal reticulocyte profile were observed in a dog with large form babesiosis presented at our institution.

The aims of this retrospective study were to determine if dogs with babesiosis and other dogs had abnormal reticulocyte profiles, and to correlate these profiles with the primary diagnosis.

Cases of large form babesiosis were identified and their reticulocyte dot plots were analyzed. Dogs with abnormal reticulocyte profiles but without microscopically apparent intraerythrocytic Babesia piroplasms were identified. The reticulocyte profiles and fluorescence ratios of dogs with and without babesiosis were compared.

Twenty of 92 dogs with babesiosis had abnormal reticulocyte profiles, including 8 with a separation between the reticulocyte and mature RBC plots or a continuum of reticulocytes from the RBC plot but with a higher density of dots in the middle of the "comet tail" than in the left quarter of the dot plot.

Thirteen of dogs without Babesia on the blood smear had abnormal reticulocyte profiles, including 3 with leukemia. The medium-fluorescence reticulocyte ratios tended to be higher in dogs with babesiosis and abnormal dot plots than in other dogs, whereas the high-fluorescence ratio was higher in one dog with leukemia. Abnormal reticulocyte dot plots and atypical reticulocyte fluorescence ratios may occur in dogs with babesiosis and alert clinical pathologists to consider this diagnosis.

Micronucleated erythrocyte frequencies in old and new world primates: Nonhuman primates are of particular relevance in evaluating the potential toxicity of drugs and environmental agents. We have used previously published information and data from the present study to establish a relationship for New World NW and Old World OW primates on the basis of the frequency of spontaneous micronucleated erythrocytes MNEs observed in peripheral blood. The results of this study indicate that NW primates have higher spontaneous MNE frequencies than OW primates, and because of this, NW primates like the common marmoset, may be suitable for evaluating the genotoxicity of chemical agents.

The water channel aquaporin-1 partitions into exosomes during reticulocyte maturation: Aquaporin-1 AQP-1 , the universal water channel, is responsible for rapid response of cell volume to changes in plasma tonicity.

In the membrane of the red cell the concentration of the protein is tightly controlled. Here, we show that AQP-1 is partially lost during in vitro maturation of mouse reticulocytes and that it is associated with exosomes, released throughout this process. AQP-1 in young reticulocytes localizes to the plasma membrane and also in endosomal compartments and exosomes, formed both in vitro and in vivo. During maturation a part of the total pool of AQP-1 is differentially sorted and released via the exosomal pathway.

A proteasome inhibitor, MG, suppresses secretion of AQP-1, implying that ubiquitination is a sorting signal for its release. We further show that modulation of medium tonicity in vitro regulates the secretion of AQP-1, thus showing that extracellular osmotic conditions can drive sorting of selected proteins by the exosomal pathway.

These results lead us to suggest that AQP-1 sorting into exosomes may be the mechanism by which the reticulocyte adapts to environmental changes during its maturation. Plasmodium falciparum can invade all stages of red blood cells, while Plasmodium vivax can invade only reticulocytes.

Both proteins possess a C-terminal hydrophobic transmembrane domain, which drives adhesion to reticulocytes. In this study, the complete genome information of the P. Eleven pvrbp family genes that included 2 pseudogenes and 9 full or partial length genes were selected and used to express recombinant proteins in a wheat germ cell-free system. The expressed proteins were used to evaluate the humoral immune response with vivax malaria patients and healthy individual serum samples by protein microarray.

The recombinant fragments of 9 PvRBP proteins were successfully expressed; the soluble proteins ranged in molecular weight from 16 to 34 kDa. The functional activity of the PvRBP family on merozoite invasion remains unknown. Micronucleation in the lens epithelium following in vivo exposure to physical and chemical mutagens. Rats were exposed to cataractogenic doses of known physical and chemical genotoxic agents in order to study the efficacy of using micronuclei to monitor mutagenicity in the lens epithelium.

The total numbers of micronuclei were counted in lens epithelia from rats exposed to graded doses of either kVp X-rays or the anti-leukemic drug, 1,4 dimethanesulfonoxybutane Myleran R. The results indicate a dose-dependent incidence of micronucleation in the lens epithelium following exposure. The findings are consistent with the hypothesis that the cataractogenicity of certain agents may be related to their effect on the genome of lens epithelial cells.

Persistence and accumulation of micronucleated hepatocytes in liver of rats after repeated administration of diethylnitrosamine.

A repeat-dose micronucleus assay in adult rat liver was recently developed [Mutat. This assay demonstrated a high detectability of hepatocarcinogens at relatively low doses, as indicated by dose-dependent micronucleus induction.

Because the adult rat liver is known to have a long life-span, this desirable property of the assay will be an advantage in detecting micronucleated hepatocytes MNHEPs that have persisted for long periods in the liver following repeated dosing. However, no data directly supporting the underlying mechanisms have been published to date.

In the present study, we verified the mechanisms by means of pulse-labeling of micronucleated hepatocytes with the thymidine analog 5-ethynyl-2'-deoxyuridine EdU. The rodent hepatocarcinogen diethylnitrosamine DEN was repeatedly administered orally to male Crl: CD SD rats 6 weeks old for up to 2 weeks, and EdU was injected intraperitoneally on days 1, 7, or Hepatocytes were isolated by use of a non-perfusion technique at 24h, 1 week, or 2 weeks after EdU injection and analyzed for EdU incorporation and micronucleus formation.

In addition, the number of terminal deoxynucleotidyl transferase-mediated nick end-labeling TUNEL -positive cells in the liver tissue increased, suggesting selective removal of micronucleated cells. Theoretical calculation of the cumulative MNHEP frequency on each of the days on which DEN was administered, taking into account the rate of loss, came out closer to the actual value observed in the liver micronucleus test.

Taken together, these results indicate that although micronucleated cells induced in rat livers by administration of the genotoxic hepatocarcinogen DEN undergo selective removal, they. Plasmodium vivax merozoites only invade reticulocytes , a minor though heterogeneous population of red blood cell precursors that can be graded by levels of transferrin receptor CD71 expression.

The development of a protocol that allows sorting reticulocytes into defined developmental stages and a robust ex vivo P vivax invasion assay has made it possible for the first time to investigate the fine-scale invasion preference of P vivax merozoites. Concurrent with these changes, clathrin pits disappear by 3 hours postinvasion, replaced by distinctive caveolae nanostructures.

These 2 hitherto unsuspected features of P vivax invasion, a narrow preference for immature reticulocytes and a rapid remodeling of the host cell, provide important insights pertinent to the pathobiology of the P vivax infection. Variation of micronucleated erythrocytes in peripheral blood of Sciurus aureogaster in relation to age: In some species, in which the human is included, the influence of age in the variation in the number of micronucleated erythrocytes MNE is known.

In the present work we show how the process of aging influences the number of spontaneous MNE in the gray squirrel Sciurus aureogaster. Because of the difficulty of knowing the age of each animal, 69 animals were weighed at their arrival to the laboratory and at the start of sample taking, with the supposition that the heaviest animals were the oldest and those with the lightest weight were the youngest.

The major number of MNE was found in the younger animals, whereas the adults displayed less MNE P Enrichment of reticulocytes from whole blood using aqueous multiphase systems of polymers. This paper demonstrates the enrichment of reticulocytes by centrifuging whole blood through aqueous multiphase systems AMPSs -immiscible phases of solutions of polymers that form step-gradients in density.

The interfaces of an AMPS concentrate cells; this concentration facilitates the extraction of blood enriched for reticulocytes.

AMPS enrich reticulocytes from blood from both healthy and hemochromatosis donors. Varying the osmolality and density of the phases of AMPS provides different levels of enrichment and yield of reticulocytes. When used on peripheral blood from normal donors, AMPS can provide a higher yield of enriched reticulocytes and a higher proportion of reticulocytes expressing CD71 than differential centrifugation followed by centrifugation over Percoll. Blood enriched for reticulocytes by AMPS could be useful for research on malaria.

Several species of malaria parasites show a preference to invade young erythrocytes and reticulocytes ; this preference complicates in vitro cultivation of these species in human blood.

that simple

As nuclear remnants are removed from rbc in the spleen. Abnormal variation in shape of rbc. Microcytic nucleated RBC with granules of iron accumulated in the mitochondria around the nuc are normally found in the bone marrow precursor to normal rbc All cases involve dysfunctional heme synthesis or processing.

Greek schistos for "divided" or schistein for "to split" injury from damaged endothelium and is a characteristic feature of microangiopathic disease of blood vessels hemolytic anemia.

AKA target cell hemoglobin E and beta thalassemia. Associated with myelophthisic anemia, commonly caused by myelofibrosis. Below 'c usually 'c for 24 months contains coagulation factors. Cryoprecipitate thawed from FFP. G6PD, PK deficiency spherocytosis sickle cell anemia. Iron def late thalassemia Lead poisoning sideroblastic anemia - dysfunctional heme synthesis. Large RBC distribution width. Since p53 plays a key role in human and mouse tumorigenesis, phaplodeficient mice are currently being evaluated as a model for assessing the carcinogenicity of perinatal exposure to NRTIs.

Manual and automated reticulocyte counts. Manual reticulocyte counts were examined under light microscopy, using the property whereby supravital stain precipitates residual ribosomal RNA versus the automated flow methods, with the suggestion that in the latter there is greater precision and an ability to determine both mature and immature reticulocyte fractions.

This work aimed to compare manual and automated methodologies for reticulocyte countings and evaluate random and systematic errors. Thus, it has been confirmed that both methods, when well conducted, can reflect precisely the reticulocyte counts for adequate clinical use. Normal and disordered reticulocyte maturation. Reticulocyte remodeling has emerged as an important model for the understanding of vesicular trafficking and selective autophagy in mammalian cells.

This review covers recent advances in our understanding of these processes in reticulocytes and the role of these processes in erythroid development. Enucleation is caused by the coalescence of vesicles at the nuclear-cytoplasmic junction and microfilament contraction. Mitochondrial elimination is achieved through selective autophagy, in which mitochondria are targeted to autophagosomes, and undergo subsequent degradation and exocytosis.

The mechanism involves an integral mitochondrial outer membrane protein and general autophagy pathways. Plasma membrane remodeling, and the elimination of certain intracellular organelles occur through the exosomal pathway. Vesicular trafficking and selective autophagy have emerged as central processes in cellular remodeling. In reticulocytes , this includes enucleation and the elimination of all membrane-bound organelles and ribosomes. Ubiquitin-like conjugation pathways, which are required for autophagy in yeast, are not essential for mitochondrial clearance in reticulocytes.

Thus, in higher eukaryotes, there appears to be redundancy between these pathways and other processes, such as vesicular nucleation.

Future studies will address the relationship between autophagy and vesicular trafficking, and the significance of both for cellular remodeling. Sensitivity of reticulocyte indices to iron therapy in an intensely training athlete.

Iron deficiency anaemia, and its debilitating effect on performance, is an area of concern for many female athletes. Automated technologies that analyse individual reticulocytes may provide a sensitive measure of bone marrow response to iron supplementation. The reticulocyte characteristics of a female volleyball player with frank iron deficiency anaemia, and her subsequent response to oral iron therapy, are reported.

Reticulocytes are immature erythrocytes; the number of reticulocytes in the peripheral blood reflects erythropoietic activity. Two cases are described to illustrate the use of the reticulocyte count in the diagnostic workup of anaemia. The first patient was a year-old woman presenting with fatigue.

Laboratory evaluation showed severe macrocytic anaemia, thrombocytopaenia and the presence of schistocytes. A low reticulocyte count suggested decreased erythropoiesis underlying the anaemia; this led to the diagnosis of vitamin B12 deficiency. The second patient, a year-old woman, also presented with fatigue and macrocytic anaemia. A high reticulocyte count indicated increased erythrocyte degradation, and the patient was eventually diagnosed with autoimmune haemolytic anaemia.

The role of reticulocytes in the differential diagnostic workup of anaemia was explored on the basis of these case descriptions. The test methodology, analytical performance, reference values and pitfalls were discussed, as well as the reticulocyte indices and their use in monitoring therapy. Reticulocytes in sports medicine: Reticulocytes are young red blood cells which develop from erythroblasts and circulate in the bloodstream for about days before maturing into erythrocytes.

With the introduction of reticulocyte count in equations and statistical models for detecting suspected blood doping, its application to sports medicine has attracted growing interest in reticulocyte behavior during training and competition seasons in athletes and experimental blood doping treatment in healthy volunteers. An update on recent publications is therefore needed to improve the interpretation of reticulocyte analysis and its variability in sportsmen.

Reticulocyte count constitutes a robust parameter during the preanalytical phase, but cell stability can be assured only if blood samples are kept at constantly cold temperatures 4 degrees C and test results will differ depending on the blood analyzer system used.

Marked intraindividual variability is the principal finding to be evaluated when exercise-induced changes are observed or illicit procedures suspected. Furthermore, reticulocyte variability is greater than that of other hematological parameters such as hemoglobin or hematocrit. Ideally, any variation should be interpreted against long-term time series for the individual athlete: Reticulocyte distribution in athletes is similar to that found in the general population, and a gender effect in some sports disciplines or selected athlete groups may be seen.

Reticulocyte variability is strongly influenced by seasonal factors linked to training and competition schedules and by the type of sports discipline. Published experimental data have confirmed the high sensitivity of reticulocyte analysis in identifying abnormal bone marrow stimulation by either erythropoietin administration or blood withdrawal and reinfusion.

Reticulocytes in untreated obstructive sleep apnoea. The short, repetitive hypoxaemic episodes observed in obstructive sleep apnoea OSA may determine small augmentations in mature red blood cells. It is unknown whether they affect reticulocyte release. This study explored whether the number and degree of maturation of circulating reticulocytes may be altered in OSA, possibly through the effect of erythropoietin. Fifty male adult patients with suspected OSA, normoxic during wakefulness, were studied.

After nocturnal polysomnography, a blood sample was withdrawn for blood cells count, erythropoietin, iron and transferrin determination. Reticulocyte concentration and degree of immaturity [high H , medium M , or low L ] were also determined. At multiple regression, only lowest nocturnal oxygen saturation remained a significant contributor to IRF r2 0. Anticlastogenic effects of galangin against mitomycin C-induced micronuclei in reticulocytes of mice.

On the other hand, when galangin was given to mice for 7 consecutive days before MMC injection, galangin showed potent anticlastogenic effects, even at the lowest dose level of 0. Results from our in vivo studies indicate that galangin is capable of suppressing the clastogenic activity of the direct acting MMC.

Together with our earlier observations, it appears that galangin is capable of protecting cells from the toxic effects of a variety of hazardous chemicals. Therefore, galangin may be an useful chemopreventive compound.

Malnutrition and infection influence the peripheral blood reticulocyte micronuclei frequency in children. Malnutrition is a serious public health problem that affects approximately one third of all children. The relationship between malnutrition and genetic damage has been widely studied in humans and animal models. The micronucleus MN assay is useful in detecting chromosome damage induced by several factors. The aim of this study was to evaluate the effects of infection and malnutrition on the frequency of MN in erythrocytes from the peripheral blood of well-nourished, uninfected WN and well-nourished, infected WNI children, and moderately malnourished UNM and severely malnourished UNS children, both with infection, using a flow cytometric analysis technique.

The percentage of reticulocytes RETs was significantly higher 1. In addition, the UNS group had a 2. These frequencies were significantly higher 1. The results suggest that infection and malnutrition induce DNA damage in children. Automated reticulocyte parameters for hereditary spherocytosis screening. The laboratory diagnosis of hereditary spherocytosis HS is based on several screening and confirmatory tests; our algorithm includes clinical features, red blood cell morphology analysis and cryohaemolysis test, and, in case of positive screening, sodium dodecyl sulphate polyacrylamide gel electrophoresis as a diagnostic test.

A total of samples were screened. Gel electrophoresis was applied to samples that were positive for the screening tests. Some other 31 anaemic conditions were also studied. In conclusion, automated reticulocyte parameters might be helpful for haemolytic anaemia diagnostic orientation even for general laboratories. In combination with cryohaemolysis, they ensure an effective and time-saving screening for HS for more specialised laboratories.

Giemsa-stained wet mount based method for reticulocyte quantification: The quantity of circulating reticulocytes is an important indicator of erythropoietic activity in response to a wide range of haematological pathologies.

While most modern laboratories use flow cytometry to quantify reticulocytes , most field laboratories still rely on 'subvital' staining. Here we demonstrate the utility of Giemsa's stain commonly used microbiology and parasitology in a 'subvital' manner to provide an accurate method to visualize and count reticulocytes in blood samples from normal and malaria-infected individuals. Micronucleated erythrocytes in newborn rats exposed to raltegravir placental transfer.

However, administration of raltegravir during pregnancy produces a compound which is transferred to high concentrations to the offspring.

The objective of this study is to evaluate the transplacental genotoxic effect of raltegravir in newborn rats. We evaluated the number of micronucleated erythrocytes MNE , micronucleated polychromatic erythrocytes MNPCE , and polychromatic erythrocytes PCE in the peripheral blood samples of the offspring of Wistar rats treated 6 days before birth with oral administration of raltegravir.

The animals were randomly assigned to five groups as follows: In addition, the effect of these drugs on the weight and height of newborns was assessed. Genotoxicity studies are required in pregnant women to determine the risk of using raltegravir to the fetuses. Reticulocytes are the youngest erythrocytes released from the bone marrow into the blood and they circulate for days before becoming mature erythrocytes.

In literature, there were studies about reticulocyte parameters that could help in differentiation of iron deficiency anemia IDA from vitamin B12 deficiency anemia. However, in those studies there were no data about differentiation of mixed anemia vitamin B12 deficiency and IDA. The purpose of this study is to explore a response to 'could reticulocyte parameters help in differential diagnosis of mixed anemia?

The study enrolled 26 patients with IDA, 22 patients with mixed anemia, 32 patients with vitamin B12 deficiency, and 32 age and gender matched healthy controls. In mixed anemia, MCV could be normal or decreased, and in peripheral blood smear erythrocytes cells could be morphologically normal.

For this reason diagnosis of mixed anemia is not easy and needs additional laboratory investigations. So, with a simple and cheap laboratory parameter, differentiation of. Acentric, autonomously replicating extrachromosomal structures called double-minute chromosomes DMs frequently mediate oncogene amplification in human tumors.

We show that DMs can be removed from the nucleus by a novel micronucleation mechanism that is initiated by budding of the nuclear membrane during S phase. DMs containing c-myc oncogenes in a colon cancer cell line localized to and replicated at the nuclear periphery.

Replication inhibitors increased micronucleation ; cell synchronization and bromodeoxyuridine—pulse labeling demonstrated de novo formation of buds and micronuclei during S phase. The frequencies of S-phase nuclear budding and micronucleation were increased dramatically in normal human cells by inactivating p53, suggesting that an S-phase function of p53 minimizes the probability of producing the broken chromosome fragments that induce budding and micronucleation.

These data have implications for understanding the behavior of acentric DNA in interphase nuclei and for developing chemotherapeutic strategies based on this new mechanism for DM elimination. Development of a technique for quantification of reticulocytes and assessment of erythrocyte regenerative capacity in birds.

To develop a reticulocyte classification scheme, optimize an avian reticulocyte staining protocol, and compare the percentages of reticulocyte types with polychromatophil percentage in blood samples from birds.

Blood samples from a red-tailed hawk and 31 ill birds. A single blood sample obtained from a red-tailed hawk Buteo jamaicensis was used to optimize the staining protocol.

For optimization of the staining protocol, 4 dilutions of whole blood with new methylene blue stain and 4 incubation times were evaluated. From samples submitted for avian CBCs, EDTA-anticoagulated whole blood samples from 31 ill birds were randomly selected and examined to compare polychromatophil and reticulocyte percentages.

Reticulocyte staining was performed in all samples by use of a 1: In Wright-Giemsa-stained blood smears, a polychromatophil percentage was similarly determined. A reticulocyte -staining protocol was optimized. Interobserver and intraobserver variations in assessment of reticulocyte and polychromatophil percentages were not significant. Results indicated that quantification of ring-form reticulocytes provides an accurate assessment of erythrocyte regenerative capacity in birds.

Flow cytometric analysis of leukocytes and reticulocytes stained with proflavine. Proflavine, an acridine analog for industrial use, was used to stain blood cells.

A drop of blood treated with ethylenediaminetetraacetic acid-2K was mixed with a 0. Leukocytes, platelets, and reticulocytes were stained but mature red blood cells were not. Chromatin in the nuclei of all leukocytes and nucleoli of lymphocytes and monocytes had greenish-yellow fluorescence, and the kind of cell could be identified by the tone and intensity of this color. Granules in granulocytes were in green. Reticular fine-granular or granulofibrous structures in the reticulocytes were brownish.

The proflavine could be used routinely in clinical laboratories because this single stain makes possible simultaneous differentiation of leukocytes and counting of reticulocytes. During the orchestrated process leading to mature erythrocytes, reticulocytes must synthesize large amounts of hemoglobin, while eliminating numerous cellular components.

Exosomes are small secreted vesicles that play an important role in this process of specific elimination. To understand the mechanisms of proteolipidic sorting leading to their biogenesis, we have explored changes in the composition of exosomes released by reticulocytes during their differentiation, in parallel to their physical properties. By combining proteomic and lipidomic approaches, we found dramatic alterations in the composition of the exosomes retrieved over the course of a 7-day in vitro differentiation protocol.

Our data support a previously proposed model, whereby in reticulocytes the biogenesis of exosomes involves several distinct mechanisms for the preferential recruitment of particular proteins and lipids and suggest that the respective prominence of those pathways changes over the course of the differentiation process. Proteolipidic composition of exosomes changes during reticulocyte maturation.

Reticulocyte quantification by flow cytometry, image analysis, and manual counting. Reticulocyte counting by flow cytometry with thiazole orange was compared to manual or automated counting of new methylene blue stained blood smears.

Forty-nine samples were compared for manual counting from randomly chosen clinical samples. Two hundred and eighty-nine samples from bone marrow transplant patients were compared during the period before and through chemo-irradiation and engraftment. The slopes of correlation plots were less than 1 when flow cytometric data were the dependent variable, suggesting that thiazole orange is less sensitive than new methylene blue.

In a third study, samples from bone marrow transplant patients were compared after increasing the thiazole orange concentration. The slopes from regression analysis were 0. This demonstrates that thiazole orange is more sensitive than new methylene blue because the window of analysis includes an increased fraction of mature reticulocytes. In addition, the precision of each assay as measured. Erroneously elevated immature reticulocyte counts in leukemic patients determined using a Sysmex XE hematology analyzer.

The immature reticulocyte fraction IRF in peripheral blood, as determined by automated reticulocyte analysis, is calculated using the sum of medium and highly fluorescent reticulocyte numbers and provides information about erythropoietic activity in bone marrow.

The purpose of this study was to investigate erroneously elevated IRF in leukemic patients, as determined using a Sysmex XE hematology analyzer Sysmex, Kobe, Japan. Normal reticulocyte scattergram patterns show regions corresponding to reticulocytes located between matured RBCs and an upper particle UPP region, which show a continuum of non-separated fraction.

The UPP represents erythroblasts and some immature reticulocytes. As a control group, peripheral blood was taken from patients with benign hematologic diseases, and their reticulocyte scattergrams all showed a normal pattern; UPP values were all less than However, the reticulocyte scattergrams of 5 of 11 leukemia patients showed abnormal patterns and displayed a gap between RBC and reticulocyte regions. For the remaining six leukemia patients with a normal reticulocyte scattergram pattern, immature reticulocytes were not markedly increased, and UPP values were less than The findings of the present study demonstrate that IRF results may be erroneously elevated in leukemia patients and indicate that hematologists should examine reticulocyte scattergrams and UPP values carefully.

Selective modulation of membrane sphingomyelin fatty acid turnover by nigericin. A study in the rat reticulocyte.

Exposure of rat reticulocytes to Nigericin produced a selective modulation of fatty acid incorporation into sphingomyelin SM of the cell membrane, via changes in SM acylation kinetics. At physiological fatty acid concentration, Nigericin accelerated 8-fold SM acylation by decreasing the apparent K m for oleate from This constitutes a novel piece of evidence for the important role of ions in SM metabolism.

Reticulocyte -derived exosomes rex are 30— nm membrane vesicles of endocytic origin released during the maturation of reticulocytes to erythrocytes upon fusion of multivesicular bodies with the plasma membrane. Proteomics analysis of rexPy confirmed their reticulocyte origin and demonstrated the presence of parasite antigens.

Our studies thus prove, for what we believe is the first time, that rex from reticulocyte -prone malarial infections are associated with splenic long-lasting memory responses. To try extrapolating these data to human infections, in vitro experiments with spleen cells of human transplantation donors were performed. Plasma-derived exosomes from vivax malaria patients exPv were actively uptaken by human splenocytes and stimulated spleen cells leading to changes in T cell subsets.

Reticulocyte -derived exosomes rex are nm membrane vesicles of endocytic origin released during the maturation of reticulocytes to erythrocytes upon fusion of multivesicular bodies with the plasma membrane.

Notably, rexPy immunization of mice induced changes in PD1 - memory T cells with effector phenotype. Further observations on the polynucleotide-induced stimulation of protein synthesis by cell-free preparations from rabbit reticulocytes. RNA stimulates the incorporation into protein of both free amino acids and of aminoacyl residues from s-RNA.

Quantitatively the response of the cell-free system to RNA is similar to that of polyuridylic acid, and there appears to be competition between messenger RNA and polyuridylic acid or polyadenylic acid. Acquisition of iron from transferrin regulates reticulocyte heme synthesis. Fe-salicylaldehyde isonicotinoylhydrazone SIH , which can donate iron to reticulocytes without transferrin as a mediator, has been utilized to test the hypothesis that the rate of iron uptake from transferrin limits the rate of heme synthesis in erythroid cells.

Taken together, these results suggest the hypothesis that some step s in the pathway of iron from extracellular transferrin to intracellular protoporphyrin limits the overall rate of heme synthesis in reticulocytes.

Spurious reticulocyte profiles in dogs with large form babesiosis: Erroneously high reticulocyte counts pseudoreticulocytosis have been reported in dogs with leukemia. Pseudoreticulocytosis and an abnormal reticulocyte profile were observed in a dog with large form babesiosis presented at our institution. The aims of this retrospective study were to determine if dogs with babesiosis and other dogs had abnormal reticulocyte profiles, and to correlate these profiles with the primary diagnosis.

Cases of large form babesiosis were identified and their reticulocyte dot plots were analyzed. Dogs with abnormal reticulocyte profiles but without microscopically apparent intraerythrocytic Babesia piroplasms were identified. The reticulocyte profiles and fluorescence ratios of dogs with and without babesiosis were compared. Twenty of 92 dogs with babesiosis had abnormal reticulocyte profiles, including 8 with a separation between the reticulocyte and mature RBC plots or a continuum of reticulocytes from the RBC plot but with a higher density of dots in the middle of the "comet tail" than in the left quarter of the dot plot.

Thirteen of dogs without Babesia on the blood smear had abnormal reticulocyte profiles, including 3 with leukemia. The medium-fluorescence reticulocyte ratios tended to be higher in dogs with babesiosis and abnormal dot plots than in other dogs, whereas the high-fluorescence ratio was higher in one dog with leukemia. Abnormal reticulocyte dot plots and atypical reticulocyte fluorescence ratios may occur in dogs with babesiosis and alert clinical pathologists to consider this diagnosis.

Micronucleated erythrocyte frequencies in old and new world primates: Nonhuman primates are of particular relevance in evaluating the potential toxicity of drugs and environmental agents. We have used previously published information and data from the present study to establish a relationship for New World NW and Old World OW primates on the basis of the frequency of spontaneous micronucleated erythrocytes MNEs observed in peripheral blood.

The results of this study indicate that NW primates have higher spontaneous MNE frequencies than OW primates, and because of this, NW primates like the common marmoset, may be suitable for evaluating the genotoxicity of chemical agents. The water channel aquaporin-1 partitions into exosomes during reticulocyte maturation: Aquaporin-1 AQP-1 , the universal water channel, is responsible for rapid response of cell volume to changes in plasma tonicity.

In the membrane of the red cell the concentration of the protein is tightly controlled. Here, we show that AQP-1 is partially lost during in vitro maturation of mouse reticulocytes and that it is associated with exosomes, released throughout this process. AQP-1 in young reticulocytes localizes to the plasma membrane and also in endosomal compartments and exosomes, formed both in vitro and in vivo.

During maturation a part of the total pool of AQP-1 is differentially sorted and released via the exosomal pathway. A proteasome inhibitor, MG, suppresses secretion of AQP-1, implying that ubiquitination is a sorting signal for its release.

We further show that modulation of medium tonicity in vitro regulates the secretion of AQP-1, thus showing that extracellular osmotic conditions can drive sorting of selected proteins by the exosomal pathway.

These results lead us to suggest that AQP-1 sorting into exosomes may be the mechanism by which the reticulocyte adapts to environmental changes during its maturation. Plasmodium falciparum can invade all stages of red blood cells, while Plasmodium vivax can invade only reticulocytes. Both proteins possess a C-terminal hydrophobic transmembrane domain, which drives adhesion to reticulocytes.

In this study, the complete genome information of the P. Eleven pvrbp family genes that included 2 pseudogenes and 9 full or partial length genes were selected and used to express recombinant proteins in a wheat germ cell-free system. The expressed proteins were used to evaluate the humoral immune response with vivax malaria patients and healthy individual serum samples by protein microarray. The recombinant fragments of 9 PvRBP proteins were successfully expressed; the soluble proteins ranged in molecular weight from 16 to 34 kDa.

The functional activity of the PvRBP family on merozoite invasion remains unknown. Micronucleation in the lens epithelium following in vivo exposure to physical and chemical mutagens. Rats were exposed to cataractogenic doses of known physical and chemical genotoxic agents in order to study the efficacy of using micronuclei to monitor mutagenicity in the lens epithelium.

The total numbers of micronuclei were counted in lens epithelia from rats exposed to graded doses of either kVp X-rays or the anti-leukemic drug, 1,4 dimethanesulfonoxybutane Myleran R. The results indicate a dose-dependent incidence of micronucleation in the lens epithelium following exposure.

The findings are consistent with the hypothesis that the cataractogenicity of certain agents may be related to their effect on the genome of lens epithelial cells. Persistence and accumulation of micronucleated hepatocytes in liver of rats after repeated administration of diethylnitrosamine. A repeat-dose micronucleus assay in adult rat liver was recently developed [Mutat. This assay demonstrated a high detectability of hepatocarcinogens at relatively low doses, as indicated by dose-dependent micronucleus induction.

Because the adult rat liver is known to have a long life-span, this desirable property of the assay will be an advantage in detecting micronucleated hepatocytes MNHEPs that have persisted for long periods in the liver following repeated dosing.

However, no data directly supporting the underlying mechanisms have been published to date. In the present study, we verified the mechanisms by means of pulse-labeling of micronucleated hepatocytes with the thymidine analog 5-ethynyl-2'-deoxyuridine EdU.

The rodent hepatocarcinogen diethylnitrosamine DEN was repeatedly administered orally to male Crl: CD SD rats 6 weeks old for up to 2 weeks, and EdU was injected intraperitoneally on days 1, 7, or Hepatocytes were isolated by use of a non-perfusion technique at 24h, 1 week, or 2 weeks after EdU injection and analyzed for EdU incorporation and micronucleus formation.

In addition, the number of terminal deoxynucleotidyl transferase-mediated nick end-labeling TUNEL -positive cells in the liver tissue increased, suggesting selective removal of micronucleated cells. Theoretical calculation of the cumulative MNHEP frequency on each of the days on which DEN was administered, taking into account the rate of loss, came out closer to the actual value observed in the liver micronucleus test.

Taken together, these results indicate that although micronucleated cells induced in rat livers by administration of the genotoxic hepatocarcinogen DEN undergo selective removal, they. Plasmodium vivax merozoites only invade reticulocytes , a minor though heterogeneous population of red blood cell precursors that can be graded by levels of transferrin receptor CD71 expression.

The development of a protocol that allows sorting reticulocytes into defined developmental stages and a robust ex vivo P vivax invasion assay has made it possible for the first time to investigate the fine-scale invasion preference of P vivax merozoites.

Concurrent with these changes, clathrin pits disappear by 3 hours postinvasion, replaced by distinctive caveolae nanostructures. These 2 hitherto unsuspected features of P vivax invasion, a narrow preference for immature reticulocytes and a rapid remodeling of the host cell, provide important insights pertinent to the pathobiology of the P vivax infection. Variation of micronucleated erythrocytes in peripheral blood of Sciurus aureogaster in relation to age: In some species, in which the human is included, the influence of age in the variation in the number of micronucleated erythrocytes MNE is known.

In the present work we show how the process of aging influences the number of spontaneous MNE in the gray squirrel Sciurus aureogaster. Because of the difficulty of knowing the age of each animal, 69 animals were weighed at their arrival to the laboratory and at the start of sample taking, with the supposition that the heaviest animals were the oldest and those with the lightest weight were the youngest.

The major number of MNE was found in the younger animals, whereas the adults displayed less MNE P Enrichment of reticulocytes from whole blood using aqueous multiphase systems of polymers. This paper demonstrates the enrichment of reticulocytes by centrifuging whole blood through aqueous multiphase systems AMPSs -immiscible phases of solutions of polymers that form step-gradients in density. The interfaces of an AMPS concentrate cells; this concentration facilitates the extraction of blood enriched for reticulocytes.

AMPS enrich reticulocytes from blood from both healthy and hemochromatosis donors. Varying the osmolality and density of the phases of AMPS provides different levels of enrichment and yield of reticulocytes.

When used on peripheral blood from normal donors, AMPS can provide a higher yield of enriched reticulocytes and a higher proportion of reticulocytes expressing CD71 than differential centrifugation followed by centrifugation over Percoll. Blood enriched for reticulocytes by AMPS could be useful for research on malaria. Several species of malaria parasites show a preference to invade young erythrocytes and reticulocytes ; this preference complicates in vitro cultivation of these species in human blood.

Plasmodium knowlesi malaria parasites invade normal human blood enriched for reticulocytes by AMPSs at a rate 2. Reticulocyte parameters of delta beta thalassaemia trait, beta thalassaemia trait and iron deficiency anaemia.

The following reticulocyte indices were assessed: The degree of anisocytosis in reticulocytes from patients with thalassaemia is correlated with HbF. For permission to use where not already granted under a licence please go to http: Dose- response relationship of temozolomide, determined by the Pig-a, comet, and micronucleus assay.

Temozolomide TMZ , a monofunctional alkylating agent, was selected as a model compound to determine its quantitative genotoxic dose- response relationship in different tissues blood, liver, and jejunum and endpoints [Pig-a-, comet-, and micronucleus assay MNT ] in male rats. TMZ was administered p. A statistically significant increase in Pig-a mutant phenotypes was observed on day 44 starting at 7.

In addition, a statistically significant increase in cytogenetic damage, as measured by micronucleated reticulocytes , was observed starting at 3. DNA strand breaks, as detected by the comet assay, showed a dose-dependent and statistically significant increase in liver, blood, and jejunum starting at doses of 3. Overall, PoD values show a high concordance between different tissues and endpoints, underlining the suitability of this experimental design to explore quantitative dose- response relationships in a variety of different tissues and endpoints, while minimizing animal use.

The reticulocyte parameters and profile provided by the GEN'S were studied in 38 anemic patients who fulfilled the diagnostic criteria for the ACD, and the results were compared with those of 38 healthy controls in a multivariate statistical analysis using the Student t-test and the receiver operating characteristics ROC curve analysis.

This study establishes the reticulocyte parameters and reticulocyte profile as provided by the GEN'S and characteristic of patients with ACD as compared with normal subjects. This should provide the basis for further studies comparing the reticulocyte profile of patients with ACD with those found in other types of anemia. The RNA in reticulocytes is not just debris: Reticulocytes contain both RNA and micro-organelles and represent the last stage of erythropoiesis before full maturation to red blood cells RBCs.

Even though there is continuing synthesis of hemoglobin and membrane-bound proteins in reticulocytes , the small amount of RNA that they contain has been regarded as non-functional residual material. Here we show that this residual RNA is both functional and essential for further reticulocyte maturation. Conversely, reticulocytes treated with an RNase Inhibitor were able to form normal biconcave cells.

Similarly, poor survival was also seen in reticulocytes in which protein synthesis had been blocked. To identify the signaling pathways involved we isolated RNAs in reticulocytes versus those present in fully matured erythroblasts cultured from hematopoietic stem cells. RNAs found in erythroblasts were related to exocytosis, metabolism, and signal transduction all of which are critical for maturation through reticulocyte and into a fully mature, biconcave erythrocyte. Our results suggest that the mRNA in reticulocytes has to be translated into novel proteins that act to preserve mitochondria and maintain cell membrane integrity as reticulocytes mature.

These results enhance our understanding of the final stage of erythropoiesis and may clarify why in vitro-generated reticulocytes for transfusion purposes survive poorly. Influence of in vitro hemolysis on nucleated red blood cells and reticulocyte counts. Nucleated red blood cells NRBCs and reticulocytes are early and important measures of red blood cells' RBCs turnover, but little is known on how spurious hemolysis may affect the reliability of these parameters.

Ten EDTA-anticoagulated samples were divided into three aliquots. The first was immediately tested, where-as the others defined A and B were mechanically hemolyzed by aspiration 5 and 10 times through a small-gauge needle. An increasing amount of hemolysis was produced in hemolyzed aliquots A and B. The RBC and reticulocyte counts progressively decreased from the nonhemolyzed sample to hemolyzed aliquots 'A' and 'B'.

The NRBC count increased in 3 of the 10 samples and decreased in the remaining seven. Hemolysis of venous blood samples may seriously jeopardize NRBC and reticulocyte counts. Reticulocyte hemoglobin equivalent to detect thalassemia and thalassemic hemoglobin variants.

Thalassemia and iron deficiency may both result in hypochromic microcytic anemia. Hematological algorithms that differentiate the two are mainly established in adult selected diagnostic groups. We aimed at creating an algorithm applicable in the presence of children, hemoglobin variants, and iron deficiency. We included reticulocyte hemoglobin equivalent, ferritin, and erythrocyte count in our algorithm. Our algorithm efficiently differentiated thalassemia and thalassemic hemoglobin variants from iron deficiency in children and adults.

Increased frequency of micronucleated exfoliated cells among humans exposed in vivo to mobile telephone radiations. The health concerns have been raised following the enormous increase in the use of wireless mobile telephones throughout the world. This investigation had been taken, with the motive to find out whether mobile phone radiations cause any in vivo effects on the frequency of micronucleated exfoliated cells in the exposed subjects.

A total of subjects including 85 regular mobile phone users exposed and 24 non-users controls had participated in this study. Exfoliated cells were obtained by swabbing the buccal-mucosa from exposed as well as sex-age-matched controls. One thousand exfoliated cells were screened from each individual for nuclear anomalies including micronuclei MN , karyolysis KL , karyorrhexis KH , broken egg BE and binucleated BN cells.

The average daily duration of exposure to mobile phone radiations is The mean of KL in controls is The value of means of KH in exposed subjects 1. Mean frequency of broken egg is found to be more in exposed subjects 0. Frequency of presence of more than one nucleus in a cell binucleated is also higher in exposed 2. Although there is a slight increase in mean frequency of KH, BE and BN in exposed subjects but the difference is not found statistically significant.

Correlation between , , and years of exposure and the frequency of MNC and TMN has been calculated and found to. Micronucleated erythrocytes in newborns of rat dams exposed to ultraviolet-A light during pregnancy; protection by ascorbic acid supplementation.

The control group was exposed to conventional light; the experimental groups were exposed to UVA nm during gestational days In some cases, ascorbic acid Asc was administered in the drinking water from gestational day 15 until delivery. Dams were sampled at h intervals during gestation, from day 16 until delivery. Analysis of the blood containing abnormal erythrocytes or giant platelets].

We have examined the influence of erythrocytes containing inclusion bodies, nucleated red cells or giant platelets on the measurement of reticulocyte count by automated machine, R Correlation of the reticulocyte count between automated and conventional method was extremely good in the blood containing red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling. However, correlation was poor when the sample contained the nucleated red cells. Reticulocyte count was decreased in the blood with significant amounts of nucleated red cells.

Since nucleated red cells themselves are not counted as reticulocytes in the machine, this was considered to be due to increased young reticulocytes which frequently appeared with nucleated red cells. Both cold agglutinated red cells and giant platelets apparently influenced the reticulocyte count by the R These results suggest that red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling do not influence the automatic counting of reticulocytes.

Although nucleated red cells, cold agglutinated red cells and giant platelets affected the reticulocyte count, the machine shows abnormal flags in most of above cases except highly agglutinated red cells , so that one can recount reticulocytes by conventional method. We conclude the machine can safely count the reticulocytes even in the blood containing abnormal red cells or platelets. Importance of spontaneous micronucleated erythrocytes in bottlenose dolphin Tursiops truncatus to marine toxicology studies.

The objective of the work was to characterize the presence of spontaneous micronucleated erythrocytes MNES from peripheral blood of bottlenose dolphins Tursiops truncatus to evaluate the possibility to use this species as potential bioindicator of genotoxic compounds.

Forty-eight blood samples from 12 bottlenose dolphins were obtain from three Mexican dolphinariums, and from 10 dolphins was possible to obtain more than one sample at different sampling times.

Smears were processed and observed with an epifluorescence microscope. No variations in the MNES values of the bottlenose dolphins that were sampled more than one occasion were found. Comparisons among dolphinariums revealed differences in MNES frequency, with the highest significant frequency observed in dolphins from dolphinarium "A" Wistar rats were exposed to 2.

Four subgroups were created in order to be irradiated 4, 16, 30 and 60 hours. Sham-exposed controls were included in the study. Animals were euthanized on the final irradiation day of each treated subgroup. Bone marrow smears were examined to determine the extent of genotoxicity after the particular treatment time. Mann-Whitney test was used for statistical evaluation of data. In comparison to the sham-exposed subgroups, the findings of polychromatic erythrocytes revealed significant differences for the 8th and 15th experimental day.

Thereafter, the frequency of micronucleated bone marrow red cells was significantly increased after 15 irradiation treatments. Comparison of micronucleus frequency data obtained after 2, 8 and 30 irradiation treatments did not reveal statistically significant differences between sham and treated subgroups.

Reticulocyte hemoglobin content predicts functional iron deficiency in hemodialysis patients receiving rHuEPO. Early detection of iron sufficiency at the level of the erythropoietic cell is necessary to optimize management of uremic anemia with recombinant human erythropoietin rHuEPO. Transferrin saturation and serum ferritin measurements have been noted to be insensitive and inaccurate measures to detect functional iron deficiency.

Recently, the reticulocyte hemoglobin content CHr has been shown to be a sensitive and specific indicator of functional iron deficiency in nondialysis patients treated with rHuEPO. In study 1, stable HD patients were studied at two outpatient dialysis centers.

CHr was normally distributed, with a mean value of CHr was weakly but consistently correlated with transferrin saturation and serum ferritin. CHr and reticulocyte number were inversely correlated with red blood cell RBC number, suggesting that the erythropoietic stimulus of routinely administered rHuEPO may have resulted in functional iron deficiency.

Month-to-month changes in CHr correlated weakly with changes in serum iron and percent transferrin saturation, but not at all with changes in serum ferritin. When we analyzed those patients with baseline CHr less than 26 pg, a level strongly suggestive of functional iron deficiency, these correlations strengthened, and in addition, month-to-month changes in CHr correlated strongly and directly with concomitant changes in RBC count, hemoglobin, and hematocrit, suggesting that rising CHr was indicative of an erythropoietic response.

In study 2, 79 patients received a single-dose infusion of mg iron dextran. After intravenous iron, CHr rose within 48 hours, peaked at 96 hours, and then fell toward. VLA-4 blockade by natalizumab inhibits sickle reticulocyte and leucocyte adhesion during simulated blood flow. Titrations with natalizumab revealed the presence of saturable levels of VLA-4 on both SCD reticulocytes and leucocytes similar to healthy subject leucocytes.

Under physiological flow conditions, the adhesion of SCD whole blood cells and isolated SCD leucocytes to immobilized vascular cell adhesion molecule 1 VCAM-1 was blocked by natalizumab in a dose-dependent manner, which correlated with cell surface receptor binding. Thus, VLA-4 blockade may be beneficial in sickle cell disease. Comparison of male versus female responses in the Pig-a mutation assay.

Labash, Carson; Avlasevich, Svetlana L. Validation of the Pig-a gene mutation assay has been based mainly on studies in male rodents. To determine if the mutagen-induced responses of the X-linked Pig-a gene differ in females compared to males, 7- or week old male and female Sprague Dawley rats were exposed to N-ethyl-N-nitrosourea ENU.

These results suggest that the slightly higher response in younger males than in the younger females may be related to differences in erythropoiesis function at that age. In conclusion, while some quantitative differences were noted, there were no. Reticulocyte and erythrocyte hypochromia markers in detection of iron deficiency in adolescent female athletes. The aim of this study was to analyse the effectiveness of new haematology parameters related to reticulocytes and mature red blood cells to differentiate pre latent and latent iron deficiency.

The study included female athletes aged years representing volleyball, handball, cycling, canoeing, cross-country skiing, swimming and judo.

To assess iron status the concentration of ferritin, soluble transferrin receptor sTfR , iron and total iron binding capacity TIBC were determined in serum.

In addition to blood morphology, the mean cellular haemoglobin content in erythrocytes CH and reticulocytes CHr , mean cellular haemoglobin concentration in reticulocytes CHCMr , the percentage of erythrocytes HYPOm and reticulocytes HYPOr with decreased cellular haemoglobin concentration, the percentage of erythrocytes LowCHm and reticulocytes LowCHr with decreased cellular haemoglobin content, and percentage of erythrocytes with decreased volume MICROm were determined.

Subjects with ferritin reticulocytes: CHCMr p Hexokinase microheterogeneity in rabbit red blood cells and its behaviour during reticulocytes maturation. Hexokinase in rabbit reticulocytes is present in two molecular forms hexokinase Ia and Ib separable by ion-exchange chromatography on DE columns.

This new hexokinase microheterogeneity is not caused by different oxidized forms of the enzyme nor influenced by the presence of proteolytic inhibitors during lysate preparation. Determine the yield of micronucleated cells in primary human fibroblasts exposed to focused soft X-rays.

Our part of the study has been to determine the effectiveness of soft X-rays at inducing chromosomal damage under conditions of direct and bystander exposure. Previous studies have been performed with primary human fibroblasts measuring micronuclei formation to determine the relative yields of direct versus bystander mediated micronuclei formation after cells were individually irradiated utilizing our novel focused soft X-ray microprobe, which is capable of producing localized submicron beams of carbon-K eV X-rays.

Only a brief overview is given here as the study has been published in several papers. Our original hypothesis was to study yields of bystander-induced micronucleated cells in both wild-type and mutant fibroblast from mouse embryo fibroblasts.

Difficulties with the level of background micronuclei in the MEFs prevented systematic studies of bystander responses in the laboratories involved in the collaboration. Our soft X-ray source has been in routine use for carbon-K X-rays and is now available with Aluminium-K 1.

A separately funded project developed a new soft X-ray microprobe which will give much greater flexibility for changing energies and giving high dose. In vivo erythrocyte micronucleus assay III. Validation and regulatory acceptance of automated scoring and the use of rat peripheral blood reticulocytes , with discussion of non-hematopoietic target cells and a single dose-level limit test.

The in vivo micronucleus assay working group of the International Workshop on Genotoxicity Testing IWGT discussed new aspects in the in vivo micronucleus MN test, including the regulatory acceptance of data derived from automated scoring, especially with regard to the use of flow cytometry, the suitability of rat peripheral blood reticulocytes to serve as the principal cell population for analysis, the establishment of in vivo MN assays in tissues other than bone marrow and blood for example liver, skin, colon, germ cells , and the biological relevance of the single-dose-level test.

Our group members agreed that flow cytometric systems to detect induction of micronucleated immature erythrocytes have advantages based on the presented data, e. Flow cytometric analysis of peripheral blood reticulocytes has the potential to allow monitoring of chromosome damage in rodents and also other species as part of routine toxicology studies. It appears that it will be applicable to humans as well, although in this case the possible confounding effects of splenic activity will need to be considered closely.

Also, the consensus of the group was that any system that meets the validation criteria recommended by the IWGT should be acceptable. A number of different flow cytometric-based micronucleus assays have been developed, but at the present time the validation data are most extensive for the flow cytometric method using anti-CD71 fluorescent staining especially in terms of inter-laboratory collaborative data. Whichever method is chosen, it is desirable that each laboratory should determine the minimum sample size required to ensure that scoring error is maintained below the level of animal-to-animal variation.

In the second IWGT, the potential to use rat peripheral blood reticulocytes as target cells for the micronucleus assay was discussed. This study assessed the effect of lead Pb exposure on the status of reticulocyte count indices in workers occupied in lead battery plants.

The present cross-sectional study was carried out on male lead battery workers. The association between BLL and reticulocyte count indices was positive and significant. Genotoxic exposure to chemical substances is common, and nursing mothers could transmit harmful substances or their metabolites to their offspring through breast milk.

We explored the possibility of determining genotoxic effects in the erythrocytes of breastfeeding rat pups whose mothers received a genotoxic compound while nursing. Ten groups of female rats and five pups per dam were studied. The control group received sterile water, and the experimental groups received one of three different doses of cyclophosphamide, colchicine, or cytosine-arabinoside.

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The disorder may be caused either by a genetic disorder or indirectly as part of myelodysplastic syndrome , [2] which can develop into hematological malignancies especially acute myeloid leukemia. Ring sideroblasts are named so because iron-laden mitochondria form a ring around the nucleus. To count a cell as a ring sideroblast, the ring must encircle a third or more of the nucleus and contain five or more iron granules, according to the WHO classification of the tumors of the hematopoietic and lymphoid tissues.

GLRX5 has also been implicated. Symptoms of sideroblastic anemia include skin paleness, fatigue, dizziness, and enlarged spleen and liver. Heart disease, liver damage, and kidney failure can result from iron buildup in these organs. Causes of sideroblastic anemia can be categorized into three groups: All cases involve dysfunctional heme synthesis or processing. This leads to granular deposition of iron in the mitochondria that form a ring around the nucleus of the developing red blood cell.

Congenital forms often present with normocytic or microcytic anemia while acquired forms of sideroblastic anemia are often normocytic or macrocytic. The anemia is moderate to severe and dimorphic.

Microscopic viewing of the red blood cells will reveal marked unequal cell size and abnormal cell shape. Basophilic stippling is marked and target cells are common. Pappenheimer bodies are present in the red blood cells. The mean cell volume is commonly decreased i. The RDW is increased with the red blood cell histogram shifted to the left. Leukocytes and platelets are normal.

Bone marrow shows erythroid hyperplasia with a maturation arrest. Serum iron , percentage saturation and ferritin are increased. The total iron-binding capacity of the cells is normal to decreased. Stainable marrow hemosiderin is increased. Echinocyte comes from the Greek word meaning "sea urchin. Heinz bodies are clumps of denatured hemoglobin resulting from exposure to high oxidant levels. As nuclear remnants are removed from rbc in the spleen. Abnormal variation in shape of rbc. Microcytic nucleated RBC with granules of iron accumulated in the mitochondria around the nuc are normally found in the bone marrow precursor to normal rbc All cases involve dysfunctional heme synthesis or processing.

Greek schistos for "divided" or schistein for "to split" injury from damaged endothelium and is a characteristic feature of microangiopathic disease of blood vessels hemolytic anemia. AKA target cell hemoglobin E and beta thalassemia.

The water channel aquaporin-1 partitions into exosomes during reticulocyte maturation: Aquaporin-1 AQP-1 , the universal water channel, is responsible for rapid response of cell volume to changes in plasma tonicity. In the membrane of the red cell the concentration of the protein is tightly controlled. Here, we show that AQP-1 is partially lost during in vitro maturation of mouse reticulocytes and that it is associated with exosomes, released throughout this process.

AQP-1 in young reticulocytes localizes to the plasma membrane and also in endosomal compartments and exosomes, formed both in vitro and in vivo. During maturation a part of the total pool of AQP-1 is differentially sorted and released via the exosomal pathway. A proteasome inhibitor, MG, suppresses secretion of AQP-1, implying that ubiquitination is a sorting signal for its release. We further show that modulation of medium tonicity in vitro regulates the secretion of AQP-1, thus showing that extracellular osmotic conditions can drive sorting of selected proteins by the exosomal pathway.

These results lead us to suggest that AQP-1 sorting into exosomes may be the mechanism by which the reticulocyte adapts to environmental changes during its maturation. Plasmodium falciparum can invade all stages of red blood cells, while Plasmodium vivax can invade only reticulocytes.

Both proteins possess a C-terminal hydrophobic transmembrane domain, which drives adhesion to reticulocytes. In this study, the complete genome information of the P. Eleven pvrbp family genes that included 2 pseudogenes and 9 full or partial length genes were selected and used to express recombinant proteins in a wheat germ cell-free system. The expressed proteins were used to evaluate the humoral immune response with vivax malaria patients and healthy individual serum samples by protein microarray.

The recombinant fragments of 9 PvRBP proteins were successfully expressed; the soluble proteins ranged in molecular weight from 16 to 34 kDa. The functional activity of the PvRBP family on merozoite invasion remains unknown. Micronucleation in the lens epithelium following in vivo exposure to physical and chemical mutagens.

Rats were exposed to cataractogenic doses of known physical and chemical genotoxic agents in order to study the efficacy of using micronuclei to monitor mutagenicity in the lens epithelium. The total numbers of micronuclei were counted in lens epithelia from rats exposed to graded doses of either kVp X-rays or the anti-leukemic drug, 1,4 dimethanesulfonoxybutane Myleran R.

The results indicate a dose-dependent incidence of micronucleation in the lens epithelium following exposure. The findings are consistent with the hypothesis that the cataractogenicity of certain agents may be related to their effect on the genome of lens epithelial cells.

Persistence and accumulation of micronucleated hepatocytes in liver of rats after repeated administration of diethylnitrosamine. A repeat-dose micronucleus assay in adult rat liver was recently developed [Mutat.

This assay demonstrated a high detectability of hepatocarcinogens at relatively low doses, as indicated by dose-dependent micronucleus induction. Because the adult rat liver is known to have a long life-span, this desirable property of the assay will be an advantage in detecting micronucleated hepatocytes MNHEPs that have persisted for long periods in the liver following repeated dosing.

However, no data directly supporting the underlying mechanisms have been published to date. In the present study, we verified the mechanisms by means of pulse-labeling of micronucleated hepatocytes with the thymidine analog 5-ethynyl-2'-deoxyuridine EdU. The rodent hepatocarcinogen diethylnitrosamine DEN was repeatedly administered orally to male Crl: CD SD rats 6 weeks old for up to 2 weeks, and EdU was injected intraperitoneally on days 1, 7, or Hepatocytes were isolated by use of a non-perfusion technique at 24h, 1 week, or 2 weeks after EdU injection and analyzed for EdU incorporation and micronucleus formation.

In addition, the number of terminal deoxynucleotidyl transferase-mediated nick end-labeling TUNEL -positive cells in the liver tissue increased, suggesting selective removal of micronucleated cells.

Theoretical calculation of the cumulative MNHEP frequency on each of the days on which DEN was administered, taking into account the rate of loss, came out closer to the actual value observed in the liver micronucleus test.

Taken together, these results indicate that although micronucleated cells induced in rat livers by administration of the genotoxic hepatocarcinogen DEN undergo selective removal, they. Plasmodium vivax merozoites only invade reticulocytes , a minor though heterogeneous population of red blood cell precursors that can be graded by levels of transferrin receptor CD71 expression.

The development of a protocol that allows sorting reticulocytes into defined developmental stages and a robust ex vivo P vivax invasion assay has made it possible for the first time to investigate the fine-scale invasion preference of P vivax merozoites.

Concurrent with these changes, clathrin pits disappear by 3 hours postinvasion, replaced by distinctive caveolae nanostructures. These 2 hitherto unsuspected features of P vivax invasion, a narrow preference for immature reticulocytes and a rapid remodeling of the host cell, provide important insights pertinent to the pathobiology of the P vivax infection.

Variation of micronucleated erythrocytes in peripheral blood of Sciurus aureogaster in relation to age: In some species, in which the human is included, the influence of age in the variation in the number of micronucleated erythrocytes MNE is known. In the present work we show how the process of aging influences the number of spontaneous MNE in the gray squirrel Sciurus aureogaster. Because of the difficulty of knowing the age of each animal, 69 animals were weighed at their arrival to the laboratory and at the start of sample taking, with the supposition that the heaviest animals were the oldest and those with the lightest weight were the youngest.

The major number of MNE was found in the younger animals, whereas the adults displayed less MNE P Enrichment of reticulocytes from whole blood using aqueous multiphase systems of polymers.

This paper demonstrates the enrichment of reticulocytes by centrifuging whole blood through aqueous multiphase systems AMPSs -immiscible phases of solutions of polymers that form step-gradients in density. The interfaces of an AMPS concentrate cells; this concentration facilitates the extraction of blood enriched for reticulocytes.

AMPS enrich reticulocytes from blood from both healthy and hemochromatosis donors. Varying the osmolality and density of the phases of AMPS provides different levels of enrichment and yield of reticulocytes. When used on peripheral blood from normal donors, AMPS can provide a higher yield of enriched reticulocytes and a higher proportion of reticulocytes expressing CD71 than differential centrifugation followed by centrifugation over Percoll. Blood enriched for reticulocytes by AMPS could be useful for research on malaria.

Several species of malaria parasites show a preference to invade young erythrocytes and reticulocytes ; this preference complicates in vitro cultivation of these species in human blood. Plasmodium knowlesi malaria parasites invade normal human blood enriched for reticulocytes by AMPSs at a rate 2. Reticulocyte parameters of delta beta thalassaemia trait, beta thalassaemia trait and iron deficiency anaemia.

The following reticulocyte indices were assessed: The degree of anisocytosis in reticulocytes from patients with thalassaemia is correlated with HbF. For permission to use where not already granted under a licence please go to http: Dose- response relationship of temozolomide, determined by the Pig-a, comet, and micronucleus assay.

Temozolomide TMZ , a monofunctional alkylating agent, was selected as a model compound to determine its quantitative genotoxic dose- response relationship in different tissues blood, liver, and jejunum and endpoints [Pig-a-, comet-, and micronucleus assay MNT ] in male rats. TMZ was administered p. A statistically significant increase in Pig-a mutant phenotypes was observed on day 44 starting at 7.

In addition, a statistically significant increase in cytogenetic damage, as measured by micronucleated reticulocytes , was observed starting at 3. DNA strand breaks, as detected by the comet assay, showed a dose-dependent and statistically significant increase in liver, blood, and jejunum starting at doses of 3.

Overall, PoD values show a high concordance between different tissues and endpoints, underlining the suitability of this experimental design to explore quantitative dose- response relationships in a variety of different tissues and endpoints, while minimizing animal use.

The reticulocyte parameters and profile provided by the GEN'S were studied in 38 anemic patients who fulfilled the diagnostic criteria for the ACD, and the results were compared with those of 38 healthy controls in a multivariate statistical analysis using the Student t-test and the receiver operating characteristics ROC curve analysis. This study establishes the reticulocyte parameters and reticulocyte profile as provided by the GEN'S and characteristic of patients with ACD as compared with normal subjects.

This should provide the basis for further studies comparing the reticulocyte profile of patients with ACD with those found in other types of anemia. The RNA in reticulocytes is not just debris: Reticulocytes contain both RNA and micro-organelles and represent the last stage of erythropoiesis before full maturation to red blood cells RBCs. Even though there is continuing synthesis of hemoglobin and membrane-bound proteins in reticulocytes , the small amount of RNA that they contain has been regarded as non-functional residual material.

Here we show that this residual RNA is both functional and essential for further reticulocyte maturation. Conversely, reticulocytes treated with an RNase Inhibitor were able to form normal biconcave cells. Similarly, poor survival was also seen in reticulocytes in which protein synthesis had been blocked. To identify the signaling pathways involved we isolated RNAs in reticulocytes versus those present in fully matured erythroblasts cultured from hematopoietic stem cells.

RNAs found in erythroblasts were related to exocytosis, metabolism, and signal transduction all of which are critical for maturation through reticulocyte and into a fully mature, biconcave erythrocyte. Our results suggest that the mRNA in reticulocytes has to be translated into novel proteins that act to preserve mitochondria and maintain cell membrane integrity as reticulocytes mature.

These results enhance our understanding of the final stage of erythropoiesis and may clarify why in vitro-generated reticulocytes for transfusion purposes survive poorly. Influence of in vitro hemolysis on nucleated red blood cells and reticulocyte counts. Nucleated red blood cells NRBCs and reticulocytes are early and important measures of red blood cells' RBCs turnover, but little is known on how spurious hemolysis may affect the reliability of these parameters.

Ten EDTA-anticoagulated samples were divided into three aliquots. The first was immediately tested, where-as the others defined A and B were mechanically hemolyzed by aspiration 5 and 10 times through a small-gauge needle. An increasing amount of hemolysis was produced in hemolyzed aliquots A and B. The RBC and reticulocyte counts progressively decreased from the nonhemolyzed sample to hemolyzed aliquots 'A' and 'B'.

The NRBC count increased in 3 of the 10 samples and decreased in the remaining seven. Hemolysis of venous blood samples may seriously jeopardize NRBC and reticulocyte counts. Reticulocyte hemoglobin equivalent to detect thalassemia and thalassemic hemoglobin variants. Thalassemia and iron deficiency may both result in hypochromic microcytic anemia.

Hematological algorithms that differentiate the two are mainly established in adult selected diagnostic groups. We aimed at creating an algorithm applicable in the presence of children, hemoglobin variants, and iron deficiency. We included reticulocyte hemoglobin equivalent, ferritin, and erythrocyte count in our algorithm. Our algorithm efficiently differentiated thalassemia and thalassemic hemoglobin variants from iron deficiency in children and adults. Increased frequency of micronucleated exfoliated cells among humans exposed in vivo to mobile telephone radiations.

The health concerns have been raised following the enormous increase in the use of wireless mobile telephones throughout the world. This investigation had been taken, with the motive to find out whether mobile phone radiations cause any in vivo effects on the frequency of micronucleated exfoliated cells in the exposed subjects.

A total of subjects including 85 regular mobile phone users exposed and 24 non-users controls had participated in this study. Exfoliated cells were obtained by swabbing the buccal-mucosa from exposed as well as sex-age-matched controls. One thousand exfoliated cells were screened from each individual for nuclear anomalies including micronuclei MN , karyolysis KL , karyorrhexis KH , broken egg BE and binucleated BN cells.

The average daily duration of exposure to mobile phone radiations is The mean of KL in controls is The value of means of KH in exposed subjects 1. Mean frequency of broken egg is found to be more in exposed subjects 0. Frequency of presence of more than one nucleus in a cell binucleated is also higher in exposed 2.

Although there is a slight increase in mean frequency of KH, BE and BN in exposed subjects but the difference is not found statistically significant.

Correlation between , , and years of exposure and the frequency of MNC and TMN has been calculated and found to. Micronucleated erythrocytes in newborns of rat dams exposed to ultraviolet-A light during pregnancy; protection by ascorbic acid supplementation. The control group was exposed to conventional light; the experimental groups were exposed to UVA nm during gestational days In some cases, ascorbic acid Asc was administered in the drinking water from gestational day 15 until delivery.

Dams were sampled at h intervals during gestation, from day 16 until delivery. Analysis of the blood containing abnormal erythrocytes or giant platelets]. We have examined the influence of erythrocytes containing inclusion bodies, nucleated red cells or giant platelets on the measurement of reticulocyte count by automated machine, R Correlation of the reticulocyte count between automated and conventional method was extremely good in the blood containing red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling.

However, correlation was poor when the sample contained the nucleated red cells. Reticulocyte count was decreased in the blood with significant amounts of nucleated red cells. Since nucleated red cells themselves are not counted as reticulocytes in the machine, this was considered to be due to increased young reticulocytes which frequently appeared with nucleated red cells. Both cold agglutinated red cells and giant platelets apparently influenced the reticulocyte count by the R These results suggest that red cells with Jolly bodies, Pappenheimer bodies or basophilic stippling do not influence the automatic counting of reticulocytes.

Although nucleated red cells, cold agglutinated red cells and giant platelets affected the reticulocyte count, the machine shows abnormal flags in most of above cases except highly agglutinated red cells , so that one can recount reticulocytes by conventional method. We conclude the machine can safely count the reticulocytes even in the blood containing abnormal red cells or platelets.

Importance of spontaneous micronucleated erythrocytes in bottlenose dolphin Tursiops truncatus to marine toxicology studies. The objective of the work was to characterize the presence of spontaneous micronucleated erythrocytes MNES from peripheral blood of bottlenose dolphins Tursiops truncatus to evaluate the possibility to use this species as potential bioindicator of genotoxic compounds.

Forty-eight blood samples from 12 bottlenose dolphins were obtain from three Mexican dolphinariums, and from 10 dolphins was possible to obtain more than one sample at different sampling times.

Smears were processed and observed with an epifluorescence microscope. No variations in the MNES values of the bottlenose dolphins that were sampled more than one occasion were found. Comparisons among dolphinariums revealed differences in MNES frequency, with the highest significant frequency observed in dolphins from dolphinarium "A" Wistar rats were exposed to 2. Four subgroups were created in order to be irradiated 4, 16, 30 and 60 hours.

Sham-exposed controls were included in the study. Animals were euthanized on the final irradiation day of each treated subgroup.

Bone marrow smears were examined to determine the extent of genotoxicity after the particular treatment time. Mann-Whitney test was used for statistical evaluation of data. In comparison to the sham-exposed subgroups, the findings of polychromatic erythrocytes revealed significant differences for the 8th and 15th experimental day. Thereafter, the frequency of micronucleated bone marrow red cells was significantly increased after 15 irradiation treatments.

Comparison of micronucleus frequency data obtained after 2, 8 and 30 irradiation treatments did not reveal statistically significant differences between sham and treated subgroups. Reticulocyte hemoglobin content predicts functional iron deficiency in hemodialysis patients receiving rHuEPO. Early detection of iron sufficiency at the level of the erythropoietic cell is necessary to optimize management of uremic anemia with recombinant human erythropoietin rHuEPO.

Transferrin saturation and serum ferritin measurements have been noted to be insensitive and inaccurate measures to detect functional iron deficiency. Recently, the reticulocyte hemoglobin content CHr has been shown to be a sensitive and specific indicator of functional iron deficiency in nondialysis patients treated with rHuEPO.

In study 1, stable HD patients were studied at two outpatient dialysis centers. CHr was normally distributed, with a mean value of CHr was weakly but consistently correlated with transferrin saturation and serum ferritin. CHr and reticulocyte number were inversely correlated with red blood cell RBC number, suggesting that the erythropoietic stimulus of routinely administered rHuEPO may have resulted in functional iron deficiency. Month-to-month changes in CHr correlated weakly with changes in serum iron and percent transferrin saturation, but not at all with changes in serum ferritin.

When we analyzed those patients with baseline CHr less than 26 pg, a level strongly suggestive of functional iron deficiency, these correlations strengthened, and in addition, month-to-month changes in CHr correlated strongly and directly with concomitant changes in RBC count, hemoglobin, and hematocrit, suggesting that rising CHr was indicative of an erythropoietic response.

In study 2, 79 patients received a single-dose infusion of mg iron dextran. After intravenous iron, CHr rose within 48 hours, peaked at 96 hours, and then fell toward.

VLA-4 blockade by natalizumab inhibits sickle reticulocyte and leucocyte adhesion during simulated blood flow. Titrations with natalizumab revealed the presence of saturable levels of VLA-4 on both SCD reticulocytes and leucocytes similar to healthy subject leucocytes. Under physiological flow conditions, the adhesion of SCD whole blood cells and isolated SCD leucocytes to immobilized vascular cell adhesion molecule 1 VCAM-1 was blocked by natalizumab in a dose-dependent manner, which correlated with cell surface receptor binding.

Thus, VLA-4 blockade may be beneficial in sickle cell disease. Comparison of male versus female responses in the Pig-a mutation assay. Labash, Carson; Avlasevich, Svetlana L. Validation of the Pig-a gene mutation assay has been based mainly on studies in male rodents. To determine if the mutagen-induced responses of the X-linked Pig-a gene differ in females compared to males, 7- or week old male and female Sprague Dawley rats were exposed to N-ethyl-N-nitrosourea ENU.

These results suggest that the slightly higher response in younger males than in the younger females may be related to differences in erythropoiesis function at that age. In conclusion, while some quantitative differences were noted, there were no. Reticulocyte and erythrocyte hypochromia markers in detection of iron deficiency in adolescent female athletes. The aim of this study was to analyse the effectiveness of new haematology parameters related to reticulocytes and mature red blood cells to differentiate pre latent and latent iron deficiency.

The study included female athletes aged years representing volleyball, handball, cycling, canoeing, cross-country skiing, swimming and judo. To assess iron status the concentration of ferritin, soluble transferrin receptor sTfR , iron and total iron binding capacity TIBC were determined in serum.

In addition to blood morphology, the mean cellular haemoglobin content in erythrocytes CH and reticulocytes CHr , mean cellular haemoglobin concentration in reticulocytes CHCMr , the percentage of erythrocytes HYPOm and reticulocytes HYPOr with decreased cellular haemoglobin concentration, the percentage of erythrocytes LowCHm and reticulocytes LowCHr with decreased cellular haemoglobin content, and percentage of erythrocytes with decreased volume MICROm were determined.

Subjects with ferritin reticulocytes: CHCMr p Hexokinase microheterogeneity in rabbit red blood cells and its behaviour during reticulocytes maturation. Hexokinase in rabbit reticulocytes is present in two molecular forms hexokinase Ia and Ib separable by ion-exchange chromatography on DE columns. This new hexokinase microheterogeneity is not caused by different oxidized forms of the enzyme nor influenced by the presence of proteolytic inhibitors during lysate preparation.

Determine the yield of micronucleated cells in primary human fibroblasts exposed to focused soft X-rays. Our part of the study has been to determine the effectiveness of soft X-rays at inducing chromosomal damage under conditions of direct and bystander exposure. Previous studies have been performed with primary human fibroblasts measuring micronuclei formation to determine the relative yields of direct versus bystander mediated micronuclei formation after cells were individually irradiated utilizing our novel focused soft X-ray microprobe, which is capable of producing localized submicron beams of carbon-K eV X-rays.

Only a brief overview is given here as the study has been published in several papers. Our original hypothesis was to study yields of bystander-induced micronucleated cells in both wild-type and mutant fibroblast from mouse embryo fibroblasts.

Difficulties with the level of background micronuclei in the MEFs prevented systematic studies of bystander responses in the laboratories involved in the collaboration. Our soft X-ray source has been in routine use for carbon-K X-rays and is now available with Aluminium-K 1. A separately funded project developed a new soft X-ray microprobe which will give much greater flexibility for changing energies and giving high dose. In vivo erythrocyte micronucleus assay III.

Validation and regulatory acceptance of automated scoring and the use of rat peripheral blood reticulocytes , with discussion of non-hematopoietic target cells and a single dose-level limit test. The in vivo micronucleus assay working group of the International Workshop on Genotoxicity Testing IWGT discussed new aspects in the in vivo micronucleus MN test, including the regulatory acceptance of data derived from automated scoring, especially with regard to the use of flow cytometry, the suitability of rat peripheral blood reticulocytes to serve as the principal cell population for analysis, the establishment of in vivo MN assays in tissues other than bone marrow and blood for example liver, skin, colon, germ cells , and the biological relevance of the single-dose-level test.

Our group members agreed that flow cytometric systems to detect induction of micronucleated immature erythrocytes have advantages based on the presented data, e. Flow cytometric analysis of peripheral blood reticulocytes has the potential to allow monitoring of chromosome damage in rodents and also other species as part of routine toxicology studies. It appears that it will be applicable to humans as well, although in this case the possible confounding effects of splenic activity will need to be considered closely.

Also, the consensus of the group was that any system that meets the validation criteria recommended by the IWGT should be acceptable. A number of different flow cytometric-based micronucleus assays have been developed, but at the present time the validation data are most extensive for the flow cytometric method using anti-CD71 fluorescent staining especially in terms of inter-laboratory collaborative data. Whichever method is chosen, it is desirable that each laboratory should determine the minimum sample size required to ensure that scoring error is maintained below the level of animal-to-animal variation.

In the second IWGT, the potential to use rat peripheral blood reticulocytes as target cells for the micronucleus assay was discussed. This study assessed the effect of lead Pb exposure on the status of reticulocyte count indices in workers occupied in lead battery plants. The present cross-sectional study was carried out on male lead battery workers. The association between BLL and reticulocyte count indices was positive and significant.

Genotoxic exposure to chemical substances is common, and nursing mothers could transmit harmful substances or their metabolites to their offspring through breast milk. We explored the possibility of determining genotoxic effects in the erythrocytes of breastfeeding rat pups whose mothers received a genotoxic compound while nursing. Ten groups of female rats and five pups per dam were studied. The control group received sterile water, and the experimental groups received one of three different doses of cyclophosphamide, colchicine, or cytosine-arabinoside.

There were increased numbers of micronucleated erythrocytes MNEs and micronucleated polychromatic erythrocytes MNPCEs in the samples from pups in the experimental groups P Micronucleated erythrocytes in newborns rats exposed to three different types of ultraviolet-A UVA lamps from commonly uses devices.

Exposure to ultraviolet-A UVA light can accidentally cause adverse effects in the skin and eyes. UVA radiation is emitted by lamps from everyday devices. In adult rats, micronucleated erythrocytes MNE are removed from the circulation by the spleen.

However, in newborn rats, MNE have been observed in peripheral blood erythrocytes. The objective of this study was to use micronucleus tests to evaluate the DNA damage caused in newborn rats exposed to UVA light from three different types of UVA lamps obtained from commonly used devices: Rat neonates were exposed to UVA lamps for 20min daily for 6days.

The neonates were sampled every third day, and the numbers of MNE and micronucleated polychromatic erythrocytes MNPCE in the peripheral blood were determined. Many hematology laboratories have adopted semi-automated digital platforms for routine use and the evidence supporting their use is increasing. The CellaVision platforms are among the most thoroughly studied digital hematology platforms; we wished to determine the accuracy of CellaVision for reticulocyte counting.

Design, Materials and Methods: We analyzed the results for pair-wise correlation and bias, and precision. The CellaVision reticulocyte counts correlated most strongly with those of the analyzer often considered the benchmark test ; the reticulocyte count distributions were noted not to be significantly different from each other across all three methods. The mean and standard deviation of bias were lowest in the comparison of CellaVision and LH counts. Our data provide additional support for the accuracy of digital hematology applications using the CellaVision DM96 platform.

Erythrocyte antigen and reticulocyte engraftment after allogeneic hematopoietic stem cell transplantation. The aim of this study was to study the usefulness of erythrocyte antigen EA measurement to study engraftment after allogeneic HSCT. Apart from the ABO group, 15 EAs representing six minor blood groups were followed by the simple tube agglutination technique.

A total of 20 The earliest donor type EA detected was from the Rh and Kidd system. These data suggest that EA and reticulocyte assays are useful in monitoring engraftment. People donating blood more than twice annually are at risk of developing iron deficiency. Little is known about the iron status of dogs enrolled in blood donor programs. Mature red blood cell RBC indices, reticulocyte indices, serum iron, serum ferritin, and total iron-binding capacity TIBC were compared between groups.

Bone marrow is the dose-limiting organ in targeted radionuclide therapy. Hence, determination of the absorbed dose to bone marrow from incorporated radionuclides is a critical element in treatment planning. This study investigated the potential of the micronucleus assay in peripheral blood reticulocytes MnRETs as an in vivo biologic dosimeter for bone marrow.

After intravenous administration of 32P-orthophosphate or 90Y-citrate in Swiss Webster mice, DNA damage induced in bone marrow erythroblastoid cells was measured by subsequent scoring of MnRETs in peripheral blood. The response to exponentially decreasing dose rates was calibrated by irradiating animals with external Cs-gamma-rays.

The maximum MnRETs frequency occurred on the second and third day after injection of 90Y-citrate and 32P-orthophosphate, respectively. The same pattern was observed for exponentially decreasing dose rates of Cs-gamma-rays.

For each type of exposure, the maximum MnRETs frequency increased in a dose-dependent manner. Using the calibrated dosimeter, the initial dose rates to the marrow per unit of injected activity were 0.

Micronuclei in peripheral blood reticulocytes can be used as a noninvasive biologic dosimeter for measuring absorbed dose rate and absorbed dose to bone marrow from incorporated radionuclides.

Age, beta thalassaemia trait, and iron-deficient anaemia significantly affect reticulocyte indices in pre-school children. Reticulocyte indices are easy to obtain, low cost parameters and have gained interest in the field of diagnosing anaemias of childhood. In the beta-thal group, HbA 2 is strongly related to all reticulocyte indices. CHr achieved the best AUC 0. Age, IDA and beta-thal significantly affect reticulocyte indices. Using various reticulocyte fractions reticulocyte protease, inhibitor-free protease, ubiquitin and inhibitor in the presence or absence of ATP, we found that the repression of an endogenous inhibitor, as suggested by others for alpha-casein proteolysis, is unlikely for bovine serum albumin.

Therefore, differences exist in the ATP-dependent proteolytic pathway of rabbit reticulocytes depending on the substrate. Fractionation of the reticulocyte ATP-dependent proteolytic system revealed at least two proteolytic and two inhibitory fractions involved in the proteolysis of bovine serum albumin.

Automated analysis of mature red blood cells and reticulocytes in SS and SC disease. Phenotypic expression of sickle cell disease SCD is highly variable. Results were compared with those of a series of patients without hematological disease. Known data were consistently confirmed, namely heterogeneity in cell volume and hemoglobin Hb concentration, as well as the premature exit of "stress" reticulocytes from the bone marrow, mostly in SS patients.

Specific changes were observed during maturation, including decreases in macrocytic and hypodense cells. Simultaneous viewing of the indices of the different RBC populations provided information on erythropoietic maturation by a rapid, reproducible, and cost-effective method.

Three procedures for determining the quantity and composition of reticulocytes visual analysis during manual microscopy; flow cytometry, semi-automatic analysis using a computerized microscopy unit are compared. The errors characteristics for each procedure and the reasons why manual analysis still remains most used are discussed. The paper points out that it is impossible to draw a conclusion on the accuracy of the procedures, by using the values of the correlation coefficient.

Evidence is provided for the thesis that the statistical accuracy may be achieved only when the image analyzer of a computerized microscopy unit is employed, therefore this procedure is recommended for use as a reference one.

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Lab or Diagnostic Findings: Lead Poisoning or Sideroblastic Anemia