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Clinical Case Reports Jun 2020Peripheral blood smear for patients with CLL and AIHA usually shows lymphoid cells with scant cytoplasm and small round nuclei with condensed chromatin, smudge cells and...
Peripheral blood smear for patients with CLL and AIHA usually shows lymphoid cells with scant cytoplasm and small round nuclei with condensed chromatin, smudge cells and spherocytes.
PubMed: 32577278
DOI: 10.1002/ccr3.2816 -
Philosophical Transactions of the Royal... Jan 2009The abundant membrane protein AE1 normally functions as an obligate anion exchanger, with classical carrier properties, in human red blood cells. Recently, four single... (Review)
Review
The abundant membrane protein AE1 normally functions as an obligate anion exchanger, with classical carrier properties, in human red blood cells. Recently, four single point mutations of hAE1 have been identified that have lost the anion exchange function, and act as non-selective monovalent cation channels, as shown in both red cell flux and oocyte expression studies. The red cell transport function shows a paradoxical temperature dependence, and is associated with spherocytic and stomatocytic red cell defects, and haemolytic anaemias. Other forms of AE1, including the native AE1 in trout red cells, and the human mutation R760Q show both channel-like and anion exchange properties. The present results point to membrane domains 9 and 10 being important in the functional modification of AE1 activity.
Topics: Anion Exchange Protein 1, Erythrocyte; Cations; Mutation; Protein Conformation
PubMed: 18957374
DOI: 10.1098/rstb.2008.0154 -
Frontiers in Physiology 2021Pyruvate kinase deficiency (PKD) is the most frequent congenital enzymatic defect of glycolysis, and one of the most common causes of hereditary non spherocytic...
Pyruvate kinase deficiency (PKD) is the most frequent congenital enzymatic defect of glycolysis, and one of the most common causes of hereditary non spherocytic hemolytic anemia. Therapeutic interventions are limited, in part because of the incomplete understanding of the molecular mechanisms that compensate for the metabolic defect. Mass spectrometry-based metabolomics analyses were performed on red blood cells (RBCs) from healthy controls (=10) and PKD patients (=5). In PKD patients, decreases in late glycolysis were accompanied by accumulation of pentose phosphate pathway (PPP) metabolites, as a function of oxidant stress to purines (increased breakdown and deamination). Markers of oxidant stress included increased levels of sulfur-containing compounds (methionine and taurine), polyamines (spermidine and spermine). Markers of hypoxia such as succinate, sphingosine 1-phosphate (S1P), and hypoxanthine were all elevated in PKD subjects. Membrane lipid oxidation and remodeling was observed in RBCs from PKD patients, as determined by increases in the levels of free (poly-/highly-unsaturated) fatty acids and acyl-carnitines. In conclusion, in the present study, we provide the first overview of RBC metabolism in patients with PKD. Though limited in scope, the study addresses the need for basic science to investigate pathologies targeting underrepresented minorities (Amish population in this study), with the ultimate goal to target treatments to health disparities.
PubMed: 34744776
DOI: 10.3389/fphys.2021.735543 -
Haematologica Sep 2020Pyruvate kinase deficiency (PKD) is the most common cause of chronic hereditary non-spherocytic hemolytic anemia and results in a broad spectrum of disease. The...
Pyruvate kinase deficiency (PKD) is the most common cause of chronic hereditary non-spherocytic hemolytic anemia and results in a broad spectrum of disease. The diagnosis of PKD requires a high index of suspicion and judicious use of laboratory tests that may not always be informative, including pyruvate kinase enzyme assay and genetic analysis of the PKLR gene. A significant minority of patients with PKD have occult mutations in non-coding regions of PKLR which are missed on standard genetic tests. The biochemical consequences of PKD result in hemolytic anemia due to red cell pyruvate and ATP deficiency while simultaneously causing increased red cell 2,3-diphosphoglycerate, which facilitates oxygen unloading. This phenomenon, in addition to numerous other factors such as genetic background and differences in splenic function result in a poor correlation between symptoms and degree of anemia from patient to patient. Red cell transfusions should, therefore, be symptom-directed and not based on a hemoglobin threshold. Patients may experience specific complications, such as paravertebral extramedullary hematopoiesis and chronic debilitating icterus, which require personalized treatment. The decision to perform splenectomy or hematopoietic stem cell transplantation is nuanced and depends on disease burden and long-term outlook given that targeted therapeutics are in development. In recognition of the complicated nature of the disease and its management and the limitations of the PKD literature, an international working group of ten PKD experts convened to better define the disease burden and manifestations. This article summarizes the conclusions of this working group and is a guide for clinicians and investigators caring for patients with PKD.
Topics: Anemia, Hemolytic, Congenital Nonspherocytic; Erythrocytes; Humans; Pyruvate Kinase; Pyruvate Metabolism, Inborn Errors
PubMed: 33054048
DOI: 10.3324/haematol.2019.240846 -
Canine Medicine and Genetics Oct 2021A 2019 ACVIM consensus statement on diagnostics for immune-mediated hemolytic anemia (IMHA) in dogs made testing recommendations. As data on the performance of...
BACKGROUND
A 2019 ACVIM consensus statement on diagnostics for immune-mediated hemolytic anemia (IMHA) in dogs made testing recommendations. As data on the performance of immunohematological tests was lacking, we undertook a comparative analysis.
MATERIAL AND METHODS
Anticoagulated blood samples from 126 dogs suspected of having IMHA submitted to a diagnostic veterinary laboratory for a routine direct antiglobulin test (DAT) and from 28 healthy control dogs were evaluated for spherocytosis and autoagglutination before and after three saline washes. Samples were also subjected to different DATs: a gel minitube and an immunochromatographic strip kit used in clinics; neutral gel column cards, microtiter plates (at 4°, 22°, and 37°C), capillary tubes, and flow cytometry used in laboratories.
RESULTS
Samples from healthy dogs yielded negative results with all immunodiagnostic tests. Among the 126 samples submitted for DAT 67 were positive by a DAT utilizing microtiter plates with goat anti-dog antiglobulin DAT at 22°C. Notably, DAT results were comparable and consistent across all evaluated methods regardless of antiglobulin and temperature used. DAT+ dogs were more severely anemic and more likely to have erythroid regeneration compared to DAT- dogs. Macroscopic agglutination in tubes or on slides was observed in 48 samples after 1:1 and 1:4 blood to saline dilution, but only persisted in four samples after washing. Among the DAT+ samples, 57% had agglutination, 87% had spherocytosis, and 45% had both. There was good correlation between spherocytosis and DAT results from the six DAT techniques, but the correlation with autoagglutination was only fair. Clinical follow-up was available for 42 dogs. Of the sample from 12 DAT+ dogs collected during treatment, 10 remained DAT+ when tested 1-24 weeks after initial assessment.
CONCLUSIONS
Based upon this comparative prospective survey, all in-clinic and laboratory DAT techniques produced similar results when performed by trained personnel and can therefore be recommended for detection of antibody-coated erythrocytes and immunohematological diagnosis. In addition, use of these tests for monitoring response of IMHA dogs to treatment might be valuable.
PubMed: 34645506
DOI: 10.1186/s40575-021-00107-0 -
Frontiers in Physiology 2014Stored blood components are a critical life-saving tool provided to patients by health services worldwide. Red cells may be stored for up to 42 days, allowing for... (Review)
Review
Stored blood components are a critical life-saving tool provided to patients by health services worldwide. Red cells may be stored for up to 42 days, allowing for efficient blood bank inventory management, but with prolonged storage comes an unwanted side-effect known as the "storage lesion", which has been implicated in poorer patient outcomes. This lesion is comprised of a number of processes that are inter-dependent. Metabolic changes include a reduction in glycolysis and ATP production after the first week of storage. This leads to an accumulation of lactate and drop in pH. Longer term damage may be done by the consequent reduction in anti-oxidant enzymes, which contributes to protein and lipid oxidation via reactive oxygen species. The oxidative damage to the cytoskeleton and membrane is involved in increased vesiculation and loss of cation gradients across the membrane. The irreversible damage caused by extensive membrane loss via vesiculation alongside dehydration is likely to result in immediate splenic sequestration of these dense, spherocytic cells. Although often overlooked in the literature, the loss of the cation gradient in stored cells will be considered in more depth in this review as well as the possible effects it may have on other elements of the storage lesion. It has now become clear that blood donors can exhibit quite large variations in the properties of their red cells, including microvesicle production and the rate of cation leak. The implications for the quality of stored red cells from such donors is discussed.
PubMed: 24987374
DOI: 10.3389/fphys.2014.00214 -
Cytometry. Part a : the Journal of the... Aug 2022The current classical blood smear technique to observe the morphology of single red blood cells (RBCs) for classification is a laborious and error-prone process. To...
The current classical blood smear technique to observe the morphology of single red blood cells (RBCs) for classification is a laborious and error-prone process. To objectively evaluate the morphology of blood cells, we established a method of computational imaging based on a programmable light emitting diode array. By using quantitative differential phase contrast (qDPC), we characterized the morphology of unlabeled RBCs as well as blood smears. By focusing on comparing the difference of imaging between unlabeled RBCs and stained RBCs under multimode microscopic imaging technology, we demonstrated that qDPC could clearly differentiate discocytes and spherocytes in both unlabeled RBCs and blood smears. The phase map provided by quantitative phase imaging further enhanced the classification accuracy. According to statistical analysis from morphological indexes, the qDPC imaging has a significantly improvement in non-circularity, texture inhomogeneity and equivalent diameters of cells. Thus, this method has a significant superiority in the capability to analyze the morphology of RBCs and could be applied to clinical assays for determining morphological, functional, and structural deterioration of RBCs.
Topics: Erythrocyte Count; Erythrocytes; Microscopy, Phase-Contrast
PubMed: 35243761
DOI: 10.1002/cyto.a.24546 -
Experimental and Therapeutic Medicine Oct 2020The aim of the present study was to enhance the understanding of the diagnosis and treatment of neonatal hereditary spherocytosis (HS). Gene sequencing and analysis was...
The aim of the present study was to enhance the understanding of the diagnosis and treatment of neonatal hereditary spherocytosis (HS). Gene sequencing and analysis was performed for the crucial splicing signals on the exons and introns of the 302 known pathogenic genes [including , and ] that are associated with this genetic deficiency of erythrocytes. A 26-day-old female presented with jaundice, anemia, an increased count in peripheral blood reticulocyte and spherocytes and a positive acidified glycerol hemolysis test. Gene sequencing revealed a novel mutation of c.3737delA (p.Lys1246fs) in the exon 16 of (14q23|NM_000347.5) gene in the patient and her father. The mutation was a frame-shifting mutation, which may result in the truncation of β-haemoglobin in the erythrocyte membrane can lead to loss of normal function, leading to the occurrence of diseases, including jaundice and hemolytic anemia. For neonates with jaundice and anemia, family history, erythrocyte index and peripheral blood smear findings have been indicated to contribute to the diagnosis of HS. In the current study, gene sequencing was indicated to be helpful for the diagnosis of HS. A novel mutation of gene was identified, which may be pathogenic via modulating the activity of β-spectrin in the erythrocyte membrane.
PubMed: 32855695
DOI: 10.3892/etm.2020.9062 -
Oxidative Medicine and Cellular... 2022Pyriproxyfen (PPF) mimics a natural hormone in insects and disrupts their growth. It is a well-known synthetic insecticide and aromatic juvenile hormone analog...
Pyriproxyfen (PPF) mimics a natural hormone in insects and disrupts their growth. It is a well-known synthetic insecticide and aromatic juvenile hormone analog frequently used in agriculture and vegetable crops to control various insect species. At present, scanty information is available about the possible potential threats of PPF in aquatic organisms. Therefore, in this study, different toxico-pathologic endpoints of PPF like DNA damage, biomarkers of oxidative stress, and status of antioxidant enzymes were determined in (freshwater fish). In our study, 60 active, free from any external obvious ailments, same size, age, and body mass were randomly allocated to four glass aquaria (T0-T3) separately containing 100 L water. The fish present in groups T1, T2, and T3 were administered PPF dissolved in water 300, 600, and 900 g/L for 30 days. Different tissues including the blood and visceral organs were obtained from each fish on days 10, 20, and 30 of the experiment. Results on various morphological and nuclear changes in red blood cells of PPF-exposed fish including pear-shaped erythrocytes, spherocytes, red blood cells with a blebbed nucleus, micronucleus, and nuclear remnants were significantly increased. Our results on genotoxicity (comet assay) recorded significantly ( ≤ 0.05) increased DNA damage in various tissues of insecticide-exposed fish. The results on oxidative stress profile (reactive oxygen species and thiobarbituric acid reactive substances) and antioxidant enzymes (reduced glutathione superoxide dismutase, peroxidase, and catalase) in multiple tissues of fish concluded significantly ( ≤ 0.05) higher quantity of biomarkers of oxidative stress and lower concentrations of different antioxidant enzymes in treated fish. Hence, the findings of our experimental research determine that PPF could induce adverse toxic impacts on multiple tissues of fish.
Topics: Animals; Antioxidants; Biomarkers; Cyprinidae; DNA Damage; Fresh Water; Gills; Insecticides; Oxidative Stress; Pyridines; Water; Water Pollutants, Chemical
PubMed: 35720182
DOI: 10.1155/2022/5859266 -
Blood Feb 1952
Topics: Erythrocyte Count; Erythrocytes; Erythrocytes, Abnormal; Humans; Spherocytes
PubMed: 14886421
DOI: No ID Found