Title: Evaluation of red blood count and biochemical parameters in elite runners - a case study Aims: The main goal of this work is to determine the dynamics of biochemical parameters and red blood count in relation to the individual phases of year training cycle in two elite runners on medium and long distances. The second goal is to examine the data with the intention of discovering the character of the trend of individual parameters in terms of four seasonal testing for each of the probands. Methods of work: he method of content analysis of record sheets related to red blood count (RBC, HGB, HCT, MCV, MCH, MCHC, RDW) and biochemical parameters (iron, transferrin, ferritin, iron saturation of transferrin) was used to develop the work. These sheets were provided to me by the coach of both probands. The provided data were recorded in tables which were the basis for the graphs in the final part of the work. The graphs were used to monitor the fluctuation of the values of individual parameters with respect to the RTC of each season, and possible mutual relations in the variability of these parameters were sought. The long-term trend was also examined in terms of the dynamics of the parameters. Results: The measured values of individual parameters of the athletes studied by me are in the range of...

Inom klinisk verksamhet är det av stor betydelse att känna till hållbarheten på olika analyter för att säkerställa att analysen inte genomförs på för gamla prover. Vid litteratursökning framgick att bland de publicerade studier som undersökt hållbarheten på blodprover har de flesta undersökt hållbarheten på venösa men inte kapillära blodprover. En vanligt förekommande analys med både venösa och kapillära prover är blodstatus, som inkluderar bland annat bestämning av hemoglobinkoncentrationen (Hb-), leukocyt- samt trombocytpartikelkoncentrationen (LPK och TPK). Analys av blodets celler kan ge allmän information om hälsotillståndet hos patienter och är därav viktiga och vanligt förekommande analyser. Syftet med examensarbetet var att undersöka hållbarheten för analyserna Hb, LPK och TPK på kapillära blodprover efter fyra och sex timmars förvaring i rumstemperatur. Provmaterialet utgjordes av 100 kapillära prover som analyserades på hematologiinstrumentet Sysmex XN-10. Analysmetoden för Hb var fotometri med natriumlaurylsulfat- (SLS-) metoden. LPK bestämdes med flödescytometri medan TPK bestämdes med impedansmetoden alternativt flödescytometri. Resultatet visade att det för Hb inte förekom någon signifikant statistisk skillnad mellan den initiala analysen och efter fyra respektive sex timmar. Analysresultatet för Hb var därmed stabilt under förhållandena i denna studie. För LPK och TPK förekom statistiskt signifikanta skillnader mellan resultatet vid den initiala analysen jämfört med efter fyra respektive sex timmar. Analysresultaten för LPK och TPK var därmed inte lika stabila som Hb är under förhållandena i studien. Den statistiska skillnaden bedömdes däremot inte ha någon klinisk betydelse, vilket ledde till slutsatsen är att kapillära prover för analys av Hb, LPK och TPK är hållbara i sex timmar vid förvaring i rumstemperatur. / One important factor in clinical practice is understanding the stability of analytes and for how long blood samples can be stored before analysis. Most published studies are based on venous blood samples rather than capillary. The knowledge about storage time for cells in capillary blood is therefore limited. Plenty of information can be obtained by analysing the blood cells and its components, including haemoglobin, white blood cells and platelets. These analyses are therefore some of the most common in clinical practice. The aim of this study was to evaluate the effect of the analysis result for haemoglobin, white blood cells and platelets in capillary blood samples after storage at room temperature for four and six hours, respectively. In the study, 100 capillary samples from anonymous patients were analysed with the haematology analyser Sysmex XN-10. The method for analysing haemoglobin was the sodium lauryl sulphate (SLS) detection method with spectrophotometry. White blood cells were analysed with fluorescens flow cytometry. Platelets were analysed with either impedance or fluorescens flow cytometry. The result of the study showed no statistically significant difference between the initial analysis results in haemoglobin and at four and six hours, concluding that within the conditions of this study, the analysis result for haemoglobin was stable. Significant statical differences were identified for both white blood cells and platelet analysis results at four and six hours. However, the identified statistical significance was not esteemed to have any clinical relevance. In conclusion haemoglobin, white blood cells and platelets in capillary blood can be stored for at least six hours at room temperature.

Complete blood count (CBC) analysis, often provided by an automated hematology analyzer, is a fundamental diagnostic test for evaluating a patient’s overall health status as a tool in diagnosing various medical conditions. CBC provides insights into the composition of the blood cells with parameters such as Mean Corpuscular Volume (MCV) and Hemoglobin (HGB). MCV represents the average size and volume of red blood cells, while HGB indicates the oxygen-carrying capacity of the blood. Different technologies are used in hematology analyzers, such as the impedance method and spectrophotometry, to achieve precise measurements of MCV and HGB. However, exploring other methodologies is of interest to potentially reduce instrument complexity and cost. Flow cytometry, based on light scatter, provides detailed information on the characteristics of individual cells and is commonly used in CBC analysis to differentiate white blood cells and reticulocytes. However, while the potential of this method for investigating MCV and HGB levels is well established, it is of significant interest to determine if the measurement techniques can be streamlined from three to one by solely using flow cytometry in this prototype analyzer. In this thesis, the possible use of measuring MCV and HGB using a flow cytometry system based on optical scatter data from a prototype hematology analyzer has been examined. The need to sphere the red blood cells before the measurements have also been investigated to evaluate reagent needs. The results have been evaluated based on the correlation factor, accuracy and precision of the proposed optical method. It is shown that the optical method in this thesis, can be used to measure MCV and HGB. However, the necessity of sphering the cells remains. Furthermore, a comparison is made between the optical method and the Sysmex XN-1000 to evaluate the accuracy of the obtained values. Finally, possible improvements and future work are suggested. / Blodstatus, utförd av automatiserade hematologiinstrument, är ett grundlägggande diagnostiskt test för att utvärdera en patients allmänna hälsotillstånd som ett verktyg för att diagnostisera olika medicinska tillsånd. Blodstatus ger inblick i blodets sammansättning med parameterar som Medelvolym av röda blodkroppar (MCV) och Hemoglobin (HGB). MCV representerar den genomsnittliga storleken och volymen av röda blodkroppar, medan HGB indikerar blodets syrebärande förmåga. Olika metoder används i hematologiinstrument, såsom impedansmetoden och spektrofotometri för att mäta MCV och HGB värden. Undersökning med hjälp av andra metoder för mätning av dessa parameter är dock utav intresse för att potentiellt minska instrument komplexitet och kostnader. Flödescytometri, baserad på ljusspridning, ger detaljerad information om egenskaperna hos enskilda celler och används ofta för att differentiera vita blodkroppar och reticulocyter. Även om potentialen att undersöka MCV- och HGB-nivåer med denna metod redan är väletablerad, är det av betydande intresse att undersöka om man kan minska mätmetoderna från tre till en, genom att enbart använda flödescytometri i detta prototypinstrument. I detta examensarbete har möjligheten att mäta MCV och HGB med hjälp av flödescytometri baserat på optisk spridningsdata från en prototyp hematologiinstrument undersökts. Behovet av att ”sfära” de röda blodkropparna innan mätningarna har också undersökts för att utvärdera reagensbehov. Resultaten har utvärderats utifrån den förslagna optiska metodens korrelationsfaktor, noggrannhet och precision. Resultatet visar att den optiska metoden som presenteras i denna uppsats, kan användas för att mäta MCV och HGB. Dock kvarstår behovet av att ”sfära” cellerna. Dessutom görs en jämförelse mellan den optiska metoden och Sysmex XN-1000 för att utvärdera noggrannheten hos de erhållna värdena. Till sist ges förslag på möjliga förbättringar och vidareutveckling av den optiska metoden för framtiden.

Some CBC parameters have been implicated in individual susceptibility to death, both in heart disease and cardiac surgery populations. The cellular elements of blood are widely affected during cardiopulmonary bypass (CPB), technique used in cardiac surgery. A Complete Blood Count called Risk Score (CBC-RS) was calculated from the average of the deviations of the various elements of the CBC and has been previously validated and published. The CBC-RS showed as excellent predictor of death from all causes in large healthy and cardiovascular risk populations. Despite the effect of CPB on the blood cells, there is no acknowledgement from the prior assessment of this score in the surgical setting. The aim of this study was to evaluate the role of CBC-RS in the surgical risk prediction (mortality and morbidity) in patients undergoing cardiac surgery with CPB. For this, it was evaluated a historical cohort of 428 patients undergoing cardiac surgery with CPB. The individual CBC-RS was calculated using the collected blood count of patients preoperatively. Logistic regression and statistical C analyzed the predictive accuracy of this score. The primary endpoint was in-hospital mortality (all-cause) and secondary outcomes included the majors and bleeding complications. In our study, CBC-RS was a predictor of hospital mortality (OR = 1.28 for each score increments, 95% CI = 1123-1458, p <0.001) and secondary outcomes (OR = 1.208, 95% CI = 1.103 to 1.323, p <0.001). The areas under the curve (AUC) was 0.697 (p <0.001) and 0.636 (p <0.001) for both the primary and secondary endpoints, respectively. In multivariate analysis, after adjustment for other risk predictors (EuroSCORE II and CPB time), the CBC-RS remained significant and was the strongest predictor of mortality. Therefore, the CBC-RS proved to be an independent predictor of mortality and surgical complications during hospitalization. It may be a useful tool in risk assessment of patients undergoing cardiac surgery. / Dentre os diversos parâmetros fornecidos pelo hemograma, alguns já foram implicados em aumento da suscetibilidade individual à morte, tanto em pacientes com patologias cardíacas quanto os submetidos à cirurgia cardíaca. Os elementos celulares do sangue são amplamente afetados durante a circulação extracorpórea (CEC), técnica usada nas cirurgias cardíacas. Um escore calculado a partir dos desvios da média dos diversos componentes do hemograma foi previamente validado, publicado e chamado de Complete Blood Count Risk Score (CBC-RS). O CBC-RS se mostrou excelente preditor de morte por todas as causas em grandes populações saudáveis ou com fatores de risco cardiovascular. Apesar do efeito da CEC sobre as células sanguíneas, não se tem conhecimento da avaliação prévia deste escore no contexto cirúrgico. O objetivo do presente trabalho foi avaliar o papel do CBC-RS na predição de risco cirúrgico (mortalidade e morbidade hospitalar) em pacientes submetidos à cirurgia cardíaca com CEC. Para isso, uma coorte histórica de 428 pacientes submetidos à cirurgia cardíaca com CEC foi avaliada. O CBC-RS individual foi calculado utilizando o hemograma coletado dos pacientes no pré-operatório. A acurácia preditora deste escore foi analisada através regressão logística e estatística C. O desfecho primário avaliado foi a mortalidade hospitalar (por todas as causas) e os desfechos secundários incluíram as complicações maiores e sangramento. Em nosso estudo, o CBC-RS foi um preditor de mortalidade hospitalar (OR = 1,28 por cada aumento de pontuação do CBC-RS, IC 95% = 1.123 - 1.458, p <0,001) e dos desfechos secundários (OR = 1,208, IC 95% = 1,103 - 1,323, p <0,001). As áreas sob a curva (AUC) foram 0,697 (p <0,001) e 0,636 (p <0,001) para os desfechos primário e secundário, respectivamente. Na análise multivariada, após ajuste para preditores de risco pré-operatório (EuroSCORE II) e transoperatório (tempo de CEC) conhecidos, o CBC-RS permaneceu significativo e foi o preditor de mortalidade mais forte. Sendo assim, o CBC-RS se mostrou um preditor independente da mortalidade e complicações cirúrgicas no período hospitalar, podendo representar uma ferramenta útil na avaliação de risco de pacientes submetidos à cirurgia cardíaca.

Fasioloides magna is a trematode parasitizing in the liver parenchyma of ruminants. Its life cycle is associated with the humid environment and includes intermediate freshwater snail hosts from family Lymnaeidae. According to the ability of host to form a certain type of a pseudocyst during fascioloidosis, they can be,divided in three groups, specific definitive hosts (red deers, fallow deers, roe deers), nonspecific definitive hosts (cattle, wild boars and elks) and atypical hosts (sheeps and goats). Beside the natural infections also the experimental infections of other potential host species has been realized (chamois, llama and bighorn sheep and traditional laboratory animals such as mice, guinea pigs, rats and rabbits). In the context of different diseases, many changes in infected organism can occur. These can be qualitatively and quantitatively evaluated. Similarly, during fascioloidosis the changes associated with the presence of the parasite in the host's body is possible to monitor, e.g. antibody production, increase in the number of eosinophils, release of eggs in faeces, internal bleeding, or the level liver damage. The liver damage is corresponding primarily to biochemical parameters of blood, not only the liver enzymes, but also other blood components, like blood proteins, lipids,...

Les facteurs régulant l’hématopoïèse en contexte de vieillissement s’avèrent incomplètement compris. Nous avons étudié les déterminants de la variabilité des traits de la formule sanguine complète dans une cohorte de 2996 femmes apparentées et non-apparentées d’ascendance française du Québec âgées de 55 à 101 ans. Les déterminants héréditaires ont été évalués par étude d’association pan-génomique. Des facteurs acquis, incluant comorbidités et hématopoïèse clonale, ont aussi été évalués. Des analyses multivariées ont été réalisées avec des modèles linéaires mixtes généralisés. Nous avons identifié des variants dans la région de GSDMA et PSMD3-CSF3 significativement associés au décompte de neutrophiles et un polymorphisme intronique à ARHGEF3 associé au décompte plaquettaire. L’effet de certains variants diminuait avec l’âge. Avec l’âge, les décomptes de neutrophiles et monocytes augmentaient tandis que le décompte des lymphocytes décroissait. Les valeurs de neutrophiles (4,1x109/L vs 3,83x109/L, valeur-p <0,001), monocytes (0,50x109/L vs 0,45x109/L, valeur-p <0,001) et plaquettes (259x109/L vs 243x109/L, valeur-p <0,001) étaient augmentées lors de comorbidités cardiométaboliques (maladie coronarienne, hypertension, diabète, dyslipidémie). L’hématopoïèse clonale ne modifiait pas les décomptes. En conclusion, nous identifions des déterminants génétiques héréditaires contribuant à la variabilité des décomptes cellulaires sanguins dans une cohorte vieillissante. De plus, le vieillissement est associé à des niveaux accrus de neutrophiles et monocytes et une diminution des lymphocytes indiquant un biais myéloïde, lequel est majoré lors de comorbidités métaboliques. L’hématopoïèse clonale ne contribue pas à ce biais myéloïde. Ces résultats supportent le fait que des facteurs extrinsèques, possiblement via un effet inflammatoire, promeuvent le biais myéloïde relié à l’âge. / Our understanding of the factors regulating peripheral blood cell traits in the setting of aging remains incomplete. We investigated the determinants underlying blood cell trait variability in a cohort of 2996 related and unrelated women of French ancestry from Québec aged 55 to 101 years. We performed a genome-wide association study to assess for genetic variants. We also assessed the impact of acquired factors such as chronic comorbidities and clonal hematopoiesis. Multivariate analyses were subsequently performed using generalized linear mixed models. We identify variants in the region of GSDMA and PSMD3-CSF3 that meet genome-wide requirements for neutrophil counts and a variant intronic to ARHGEF3 for platelet counts. With aging, the effect of certain variants decreased. Aging was associated with increasing neutrophil and monocyte counts and decreasing lymphocyte counts. We also document that individuals with cardiometabolic comorbidities (diabetes, coronary heart disease, hypertension and dyslipidemia) exhibit significantly higher neutrophil (4.1x109/L vs 3.83x109/L, p-value <0.001), monocyte (0.50x109/L vs 0.45x109/L p-value <0.001), and platelet (259x109/L vs 243x109/L, p-value <0.001) counts. Clonal hematopoiesis did not contribute significantly to these traits. In conclusion, germline variants related to GSDMA and PSMD3-CSF3 contribute to neutrophil counts and a SNP intronic to ARHGEF3 contributes to platelet counts. Aging is associated with a myeloid shift with increased levels of neutrophils and monocytes, and reduced lymphocyte counts. This myeloid-biased skewing is further increased with cardiometabolic comorbidities. Clonal hematopoiesis does not contribute to this phenomenon. These findings support that cellextrinsic factors may contribute to the myeloid shift possibly through low-grade inflammation.