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Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study
Zubatá, Karolína ; Čečková, Martina (advisor) ; Hronek, Miloslav (referee)
Charles University University of Porto Faculty of Pharmacy in Hradec Králové Faculty of Pharmacy Department of Pharmacology and Toxicology Department of Biological Sciences Student: Karolína Zubatá Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Consultants: Susana Rocha, Ph.D., prof. Alice Santos-Silva, Ph.D. Title of diploma thesis: Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study Increased levels of oxidative stress (OS) have been described in healthy, full-term newborns as a consequence of the drastic changes introduced by birth and by the exposure to extrauterine environment. Our intention was to examine the OS levels in red blood cells (RBCs) of neonates and to further understand the changes that the newborn organism undergoes with its newly- acquired autonomy as this knowledge is limited and there are no reference values. Umbilical cord blood samples were collected from a small population of newborns (n = 8) and several hematological and biochemistry parameters were evaluated. Our experimental data consist of OS biomarkers measurements performed in different fractions of blood (RBC membrane, total RBCs and plasma): membrane bound hemoglobin (MBH), lipid peroxidation (LPO), quantification of catalase (CAT) and glutathione peroxidase (GPx) activities,...
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Morphological changes of erythrocytes in selected diseases
RIEGEROVÁ, Jana
This bachelor's thesis deals with the morphology of ecrocytes and their anomalies related to selected diseases. The theoretical part of the thesis is focused on the description of individual ecrocyte morphological abnormalities and the description of selected diseases for which morphological transformations are typical. It also aims at the explanation of their importance for diagnosis and disease progress. The practical part comprises description of blood smears microscope evaluation procedure, evaluation of erytrocyte morphology and assessment of morphological transformations in regard to patients' diagnoses. More than 80 samples were evaluated, all of which were collected between 2nd May 2017 and 25th May 2017 at the Department of Hematology in Písek. Selected blood smears containing morphological anomalies. Samples were classified in nineteen categories according to the diagnoses. Afterwards, morphological transformations were evaluated in each category. Frequency of identified abnormalities was assessed and then also their possible impact on the disease was considered. Among three most frequently detected erytrocyte morphological transformations belong polychromasia in 45 samples, anisocytosis in 43 samples and ovalocytosis in 22 samples. The observed morphological transformations were in some cases typical for the basic diagnosis and treatment, in some cases the detected morphology was untypical for diagnosed disease and informed the physician of another possible health issue. Patients with untypical signs in blood smears were consequently subjected to additional tests, which revealed the etiopathogenesis of such transformations. The results thus demonstrate the importance of blood smear evaluation for making a precise diagnosis.
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Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study
Zubatá, Karolína ; Čečková, Martina (advisor) ; Hronek, Miloslav (referee)
Charles University University of Porto Faculty of Pharmacy in Hradec Králové Faculty of Pharmacy Department of Pharmacology and Toxicology Department of Biological Sciences Student: Karolína Zubatá Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Consultants: Susana Rocha, Ph.D., prof. Alice Santos-Silva, Ph.D. Title of diploma thesis: Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study Increased levels of oxidative stress (OS) have been described in healthy, full-term newborns as a consequence of the drastic changes introduced by birth and by the exposure to extrauterine environment. Our intention was to examine the OS levels in red blood cells (RBCs) of neonates and to further understand the changes that the newborn organism undergoes with its newly- acquired autonomy as this knowledge is limited and there are no reference values. Umbilical cord blood samples were collected from a small population of newborns (n = 8) and several hematological and biochemistry parameters were evaluated. Our experimental data consist of OS biomarkers measurements performed in different fractions of blood (RBC membrane, total RBCs and plasma): membrane bound hemoglobin (MBH), lipid peroxidation (LPO), quantification of catalase (CAT) and glutathione peroxidase (GPx) activities,...
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Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study
Zubatá, Karolína ; Čečková, Martina (advisor) ; Hronek, Miloslav (referee)
Charles University University of Porto Faculty of Pharmacy in Hradec Králové Faculty of Pharmacy Department of Pharmacology and Toxicology Department of Biological Sciences Student: Karolína Zubatá Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Consultants: Susana Rocha, Ph.D., prof. Alice Santos-Silva, Ph.D. Title of diploma thesis: Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study Increased levels of oxidative stress (OS) have been described in healthy, full-term newborns as a consequence of the drastic changes introduced by birth and by the exposure to extrauterine environment. Our intention was to examine the OS levels in red blood cells (RBCs) of neonates and to further understand the changes that the newborn organism undergoes with its newly- acquired autonomy as this knowledge is limited and there are no reference values. Umbilical cord blood samples were collected from a small population of newborns (n = 8) and several hematological and biochemistry parameters were evaluated. Our experimental data consist of OS biomarkers measurements performed in different fractions of blood (RBC membrane, total RBCs and plasma): membrane bound hemoglobin (MBH), lipid peroxidation (LPO), quantification of catalase (CAT) and glutathione peroxidase (GPx) activities,...
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Effect of parasites on haematological traits in vertebrates
Vacková, Julie ; Vinkler, Michal (advisor) ; Svobodová, Jana (referee)
Deviation out of homeostasis is reflected by physiological stress and altered immunological activity leading to changes in blood composition. Haematological examination may reveal deflection from the long-term optimum-the state of health. Haematological changes associated with parasitism are in some cases so distinctive that they allow the estimation of the parasite type affecting the host health. I aim to summarise basic methods utilised in veterinary haematology and evaluate the influence of individual groups of parasites on haematological parameters related to red and white blood cells. In vertebrates the most significant haematological changes are caused by multicellular endoparasites. Typical is eosinophilia and increased total leucocyte count. Ectoparasites are usually responsible for decrease in erythrocytes and increase in leucocytes. Extracellular bacteria increase total neutrophil and lymphocyte counts while intracellular bacteria cause decrease in all blood cell types. Viruses often decrease number of erythrocytes and leucocytes and change neutrophil/lymphocyte ratio. Changes in the haematological parameters caused by protists are difficult to generalize and not much is presently known about the influence of fungi on the blood composition in vertebrates. Variation in the effect of...
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Vztahy mezi úrovní ploidie, velikostí genomu a velikostí buňky v sérii modelů ryb ploidní úrovně od 2n do 14n
BYTYUTSKYY, Dmytro
The ploidy level of diploid and induced triploid tench, Tinca tinca, was verified using flow cytometry to determine relative DNA content of 4',6-diamidino-2-phenylindole (DAPI)-stained erythrocyte nuclei. The C-value (haploid nuclear DNA content; pgDNA.nucleus-1) of these same individuals was determined by means of Feulgen image analysis densitometry, in comparison to the chicken standard (Gallus gallus domesticus; 1.25 pgDNA.nucleus-1, P < 0.05), using three different approaches. Highly similar mean C-values were obtained, thus confirming the possibility of using tench blood as standard in European pond aquaculture for ploidy and DNA content determination in fishes. Feulgen image analysis densitometry (FIAD), flow cytometry (FC) and confocal laser scanning microscopy (CLSM) were used to study the relationship between the DNA content (pgDNA.nucleus-1), nuclear area (?m2), nuclear volume (?m3) and 3-D structure of erythrocyte nuclei in a series of fish ploidy level models: diploid tench (Tinca tinca) (2n), Cuban gar (Atractosteus tristoechus) (2n), triploid tench (3n), evolutionary tetraploid sterlet (Acipenser ruthenus) and stellate sturgeons (A. stellatus) (4n), evolutionary octaploid Siberian sturgeon (A. baerii) and Russian sturgeon (A. gueldenstaedtii) (8n), spontaneous triploid Siberian and Russian sturgeons exhibiting dodecaploidy (12n), evolutionary 12n shortnose sturgeon (A. brevirostrum), and experimentally obtained sturgeon hybrids that were tetraploid, hexaploid (6n), heptaploid (7n), octaploid (8n), decaploid (10n), dodecaploid (12n) and/or tetradecaploid (14n). Standards used for FIA were blood smears of chicken (2.5 pgDNA.nucleus-1), diploid and induced triploid tench (2.04 and 3.1 pgDNA.nucleus-1, respectively). All ploidy levels were first verified by means of FC. Increase in ploidy was accompanied by growth of the nucleus and an increase in the number of flattened ellipsoid nuclei with increased transverse diameter. The volume (Vvoxel) of erythrocyte nuclei, as the sum of voxels calculated from live cells, seems more accurate than volume (Vaxis) calculated from measuring the major and minor axis, especially at higher and odd ploidy levels. Data of absolute and relative DNA content were in agreement with previously published reports. Species of the same ploidy level, however differing in their DNA content, exhibited a similar mean erythrocyte nuclear area, as could be demonstrated on A. ruthenusand and A. stellatus (19.27 and 19.79 ?m2 with a respective mean DNA content of 3.72 and 4.68 pgDNA.nucleus-1) and volume as could be demonstrated on a A. ruthenus and hybrid of A. ruthenus and H. huso(48.3 and 48.9 ?m3 with a respective mean DNA content of 3.74 and 3.10 pg DNA.nucleus-1). Similar relationship was found for the ploidy 6n, 8n, 10n, 12n. The 0.46-1.58 pgDNA increments in DNA content of erythrocytes thus had no effect on their nuclear area/volume. With increasing ploidy level, the DNA concent ration (pgDNA per 1 ?m3 of erythrocyte nuclear volume) as well as surface-to-volume ratio was found not to increase linearly. Nuclear DNA content appeared to be more condensed with an increase of the ploidy level. Observed results deduce properties of whole cell and particularly of the nuclei in series of ploidy levels fishes, adding conformations of nucleotypic hypothesis in context of cell/nuclear size and genome size relationships, as well as taxonomic position of sturgeons.
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Effect reduced levels of vitamin B12 and folic acid on the morfology of erythrocytes
POPELOVÁ, Michaela
The morphology of erythrocyte is an important diagnostic element. Changes in morphology can point to certain diseases. Among various morphological features of erythrocytes the automatic analyzers commonly examine their size and stainability. The size of erythrocyte is ranging between 6.7 and 7.7 mmicrometrů. The smaller erythrocytes are called microcytes while bigger ones are called macrocytes. The size of erythrocytes is assessed by MCV (Mean Cell Volume), which indicates the average corpuscular volume. The supply of substances such as proteins, vitamins and minerals is important for proper development and metabolism of erythrocytes. Especially iron, vitamin B12 and folic acid are substances that affect the morphology of the erythrocyte. Deficiency of these substances leads to morphological changes in the blood picture. The deficiency of iron is presented in the peripheral blood like microcytosis (MCV <80 fl). The macrocytosis is the opposite change, which means deficiency of vitamin B12 and folic acid (MCV> 97 fl). This change is the main theme of project. As a result of deficiency of vitamin B12 and folic acid, changes in bone marrow are formed in addition to changes observed in the peripheral blood. Deficiency of vitamin B12 causes the megaloblastic reconstruction of bone marrow. This reconstructed bone marrow differs from normoblastic bone marrow by size, shape and and the gap between the cytoplasm and nucleus maturation. The retardation of nuclear maturation is caused by a suppressed formation of DNA related to the above mentioned deficiency of vitamin B12. Megaloblastic anemia and pernicious anemia are other hematological diseases caused by deficiency of vitamin B12. These diseases are at work also mentioned. The causes, which lead to a lack of both vitamins, are variable, for instance their insufficient food intake, or a defect in their assimilation. At present automatic analyzers offer a full range of tested parameters of blood cells. The samples were examined by an automatic analyzer Coulter LH 750, which operates on the principle of impedance. The value of MCV analyzer calculates using the software as the average of all measured impedance pulses. The macrocytosis was presented in 4% of the total number of samples examined for levels of vitamin B12. In 52% of samples deficient values of B12 (< 150 ng/l) content were observed. The macrocytosis was present in 7.5% of the total number of samples examined for level of folic acid. The deficit values of folic acid (< 3,1 microgram/l) were observed in 38% of all cases. At increased values of MCV 27% deficiency of vitamin B12 and 33% deficiency of folic acid was found. This work confirmed the link between lack of vitamin B12 and folic acid and the subsequent development of morphological changes. The deficiency in vitamin B12, in folic acid or in both vitamins occurred at high MCV.
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