L’AE1 (SLC4A1) est un échangeur chlorure/bicarbonate. Cette protéine est la protéine membranaire la plus abondante à la surface des globules rouges des vertébrés. Elle est participe au transport du CO2 et à l’ancrage du cytosquelette à la membrane plasmique. Des mutations ponctuelles dans la partie membranaire de l’AE1, liées à des pathologies humaines, convertissent l’échange électroneutre en voie de conductance pour le sodium et le potassium ou induisent une fuite de cations dans un échangeur d’anions toujours fonctionnel. Les déterminants moléculaires qui induisent les mouvements d’ions au travers de cet échangeur sont encore inconnus. Le travail présenté a eu pour but d’identifier et de cartographier le site de transport de la protéine normale ou « pathologique ». Nous avons adapté à l’AE1 des outils basés sur la chimie des sulfhydriles capable de donner des informations sur le rôle d’acides aminés dans le site de transport de la protéine. Cette stratégie combinée à l’élaboration d’un modèle tridimensionnel de la protéine in silico basé sur le symporteur uracile/proton nous a permis de définir le site de transport de l’AE1. Nos résultats démontrent qu’un site de transport unique dans l’AE1 peut basculer entre 3 conformations différentes : échange chlorure/bicarbonate, fuite de cation et échange anionique ou fuite de cation uniquement. Ce site met en jeu les segments transmembranaires (TM) 3, 5 et 8 ainsi qu’une boucle intracellulaire très conservée située entre les TM 8 et 9. Le site de transport se structure autour des acides aminés L468, F471, L530, I533 et L673 se terminant au niveau du E681. La boucle intracellulaire 690 à 705 agissant comme un filtre à cations. / AE1 (SLC4A1, band 3) is a member of the SLC4 bicarbonate transporter family. This protein is the most abundant membrane protein on the surface of vertebrate red blood cells. The AE1 exchanges chloride and bicarbonate ions in physiological conditions. In red blood cells, it is essential to many tasks including CO2 transport and cytoskeleton anchoring in the plasma membrane. Point mutations in the membrane spanning domain of AE1 convert the electroneutral exchange into a conductance for sodium and potassium cations or induce a cation leak in a still functional anionic exchanger.The molecular determinants that induce the movement of ions through the exchanger are still unknown. This work aims at identifying and mapping the transport site of AE1 protein in normal and pathological conditions. We modified a sulfhydryl-based chemistry to AE1. This provided information on the role of amino acids in the transport site of the protein. This strategy combined with the development of a three-dimensional model of the protein in silico, based on the uracil/proton symporter, allowed us to define the transport site of AE1. Analysis of our results showed that a single transport site in AE1 can switch between three different conformations depending on protein mutation: classical chloride/bicarbonate exchange, cation leak and anion exchange, cation leak only. The transport site involves the transmembrane segments 3, 5 and 8 and a highly conserved intracellular loop between transmembrane segments 8 and 9. The transport site is centered around the amino acids L468, F471, L530, L673, I533 and ends at glutamic acid 681. The intracellular loop 690-705 acts as a cation filter.

O grau de hemólise (GH) é um dos parâmetros de qualidade de concentrados de hemácias (CH). Conforme a Portaria 158/2016, ao menos 1% da produção mensal de CH deve ser controlada para o GH e 75% desta parcela deve apresentar resultado inferior a 0,8% de hemólise em relação à massa eritrocitária no último dia de validade do CH. O GH é definido como a porcentagem de hemoglobina livre (HbL) em relação à hemoglobina total (HbT) com a devida correção do volume globular do CH. O método analítico utilizado para a dosagem da HbL pela maioria dos hemocentros da rede pública nacional não referencia a fonte bibliográfica consultada. A hemoglobina liberada dos eritrócitos devido à hemólise é tóxica e desencadeia reações fisiopatológicas, resultando em implicações clínicas com gravidade que varia em função do grau de hemólise e do volume de hemácias transfundidas. O objetivo deste estudo foi avaliar os resultados de três metodologias analíticas espectrofotométricas para a determinação da HbL. Foram comparados o método de espectrofotometria direta de Harboe, o método de espectrofotometria direta de Cinco comprimentos de onda (5CO) e o método espectrofotométrico de Primeira derivada (1ªD). Os métodos de Harboe e de 5CO utilizam fórmulas matemáticas que convertem diretamente as absorbâncias lidas no espectrofotômetro UV/Visível em concentração de HbL. O método de 1ªD requer espectrofotômetro de varredura para a visualização dos espectros e a concentração da HbL é dada pelo valor referente à distância entre o vale e o pico de absorção da hemoglobina e do fator de correção, resultante de curva de calibração. Nesse estudo foram testados os sobrenadantes de 187 CH com CPDA-1 e CH com solução aditiva de SAG-Manitol. As amostras foram diluídas segundo o aspecto visual de hemólise do sobrenadante. A cada corrida analítica foram incluídas amostras controle preparadas in-house a partir de CH com concentração de HbT conhecida. O método de Harboe emprega leituras espectrofotométricas em 380, 415 e 450 nm. Para o método de 5 CO as absorbâncias são lidas em 370, 415, 510, 577 e 600 nm. O método da 1ªD utiliza o espectro de absorção analisado em 568 nm e 580 nm. A correlação entre as três metodologias testadas foi considerada ótima, evidenciando a equivalência entre os métodos. O método de Harboe mostrou-se compatível para a dosagem de baixas e de altas concentrações de HbL. O intervalo de linearidade espectrofotométrica oscilou de 0,00041 a 0,06075 g/dL. Entretanto, para resultados confiáveis de HbL, é imprescindível que o espectrofotômetro tenha especificação de largura da banda espectral igual ou inferior a 5 nm. O método de Harboe é embasado cientificamente, de fácil execução e baixo custo. Este método proporciona resultados fidedignos, reprodutíveis e padronizados, além de dispensar o uso de substâncias químicas perigosas. Complementarmente, disponibilizou-se um Procedimento Operacional Padrão para a determinação da HbL pelo método de Harboe e um Guia visual de hemólise para assessorar os profissionais que atuam na área de controle de qualidade de hemocomponentes em hemocentros nacionais. / The hemolysis is one of the parameters of the red cell concentrate (RCC) control quality. According to the Brazilian Ordinance 158/2016, at least 1% of the monthly production of RCC should be controlled regarding hemolysis and 75% of this amount should present below than 0.8% of hemolysis in relation to the red cell mass at end of RCC storage. The hemolysis is defined as the percentage of free hemoglobin (FHb), relative to the total hemoglobin (THb), with the appropriate correction of the RCC hematocrit. The analytical method used by most the national public healthcare blood centers to dosage FHb does not reference the bibliography that has been consulted. Hemoglobin released from the erythrocytes due to hemolysis is toxic and triggers pathophysiological reactions, resulting in clinical implications whose severity varies depending on the hemolysis grade and the amount of transfused RCC. The aim of this study was to evaluate the results of three spectrophotometric analytical methodologies for the determination of FHb. The Harboe spectrophotometry method, the method of five wavelengths direct spectrophotometry (5Wa) and the first derivative spectrophotometric method (1stD) were compared to each other. The Harboe and the 5Wa methods use mathematical formulas that directly convert the absorbance read from the UV/visible spectrophotometer in FHb concentration. The 1stD method requires scanning spectrophotometer to visualize the spectra and concentration of the FHb is given by the value calculated from the distance between the valley and the peak absorption of hemoglobin and a correction factor, resulting from the calibration curve. One hundred eighty-seven (187) RCC supernatants with CPDA-1 and RCC with SAG-mannitol additive solution were tested in this study. The samples were diluted according to the visual appearance of supernatants hemolysis. For each analytical run in-house control samples, prepared from RCC with known THb concentration, were included. The Harboe method employs spectrophotometric readings at 380, 415 and 450 nm. Absorbance is read at 370, 415, 510, 577 and 600 nm with the 5 Wa method. In the 1stD method the absorption spectrum is analyzed at 568 and 580 nm. There was correlation between the three tested methodologies that were tested, demonstrating equivalence between the methods. The Harboe method was compatible for dosage of low and high FHb concentrations. The spectrophotometer linear range varied from 0.00041 to 0.06075 g/dL. However, in order to achieve reliable FHb dosages it is imperative that the spectrophotometer has a spectral bandwidth equal to 5 nm or below. The Harboe method is scientifically based, easy to perform and inexpensive. It provides reliable, reproducible and standardized results, dismissing the use of dangerous chemical substances. In addition, this study has provided an Operational Procedure to determine FHb by the Harboe method and a Visual Guide of hemolysis to assist professionals working in the quality control of blood products.

A deformabilidade é a característica que permite ao eritrócito normal de 7 a 8 micrômetros (m) circular por capilares de até 3m de diâmetro. Esse fenômeno depende da geometria celular, da viscosidade interna e das propriedades visco-elásticas da membrana eritrocitária. Dentre as várias técnicas de estudo da deformabilidade eritrocitária (DE), como: a aspiração por micropipeta, a filtração e a reoscopia, destaca-se a ectacitometria. Essa técnica utiliza um viscosímetro de fluxo laminar onde as modificações de forma dos eritrócitos são monitoradas continuamente por um feixe de raio LASER, processadas por microcomputador e inseridas em gráfico para posterior análise. A ectacitometria fornece o Índice de Deformabilidade (ID), o qual proporciona a medida da eliptocitogênese dos eritrócitos quando submetidos a uma força denominada shear stress. A anemia ferropriva é uma patologia muito freqüente na prática médica. Apresenta anormalidades morfológicas expressivas como: microcitose, hipocromia, ovalócitos, eliptócitos e hemácias em alvo. Alterações de deformabilidade eritrocitária foram descritas em diversas situações como na esferocitose hereditária, eliptocitose hereditária e anemias hemolíticas auto-imune. Na anemia ferropriva os trabalhos de deformabilidade eritrocitária são controversos. O presente estudo avalia a DE, utilizando a técnica da ectacitometria, em 21 pacientes portadores de anemia ferropriva documentada, antes e depois do tratamento com sais de ferro. Embora o tratamento da anemia tenha sido eficaz (Hb antes- 8,52 g/dl e Hb depois -12,74 g/dl), alguns pacientes persistiram com morfologia eritrocitária alterada. Os resultados demonstram DE diminuída em pacientes portadores de anemia ferropriva, quando comparada ao grupo controle (p< 0,0007). A ausência de normalização e manutenção da diferença estatística após a terapêutica (p< 0,03), em baixos valores de shear stress, pode ser atribuida à manutenção das alterações morfológicas eritrocitárias. Não foi verificada correlação entre o grau da anemia e a redução da DE. A diminuição da DE apresenta maior correlação com a microcitose, sendo que a hipocromia parece não interferir de maneira importante. A concomitância dos dois fatores pode somar ou anular os seus efeitos sobre a DE. O presente estudo sugere que o fator responsável pela diminuição da DE na anemia ferropriva é a microcitose. Recentemente, relatos de anemia ferropriva associada a fenômenos trombóticos aumentaram o interesse no estudo da DE para melhor compreensão desses casos. / The deformability allows the 7 to 8 cm red cell to circulate through capillaries of 3 cm. This phenomenon depends on cellular geometry, internal viscosity and viscoelastic properties of the membrane. Among the various techniques of erytrocyte deformability (ED) analysis such as: micropipette aspiration, filtration and reoscopy, we chose ektacytometry. This technique uses a laminar flow viscometry, where red cell shape changes are continuously monitorated by LASER, processed by a computer and inserted in a graphic for further analysis. Ektacytometry measures the Deformability Index (DI), which shows the size of eliptocytogenesis of the eritrocyte under shear stress force. Iron deficiency anemia is a very frequent disease in medical practice. It presents expressive morphologic alterations such as microcytosis, hypocromy, ovalocytosis, eliptocytosis and target cells. Erytrocyte deformability has been described in a number of situations like hereditary spherocytosis, hereditary elitptocytosis and autoimmune hemolytic anemia. Concerning iron deficiency anemia, authors are controversial. The present study evaluates erytrocyte deformability, using ectacytometry in 21 patients carrying documented iron deficiency before and after therapeutics with iron components. Although the anemia treatment proved to be efficient (before Hb- 8,52 g/dl and after Hb- 12,74 g/dl), some patients persisted with erytrocyte alteration morfology. Results demonstrate diminished erytrocyte deformability in people with iron deficiency anemia, when compared with the control group (p< 0,0007). The absence of regularization and maintenance of statistical difference after treatment (p< 0,03) in low shear stress can be attributed to the persistence of red cell anomalies. There is no relation between the level of anemia and reduced ED. The erytrocyte deformability diminished is greatly related to microcitosys, even if hipocromy seems to not interefere importantly. The two factors altogether can either sum or nulify the effects over erytrocyte deformability. The present study suggests that the responsible factor for diminished erytrocyte deformability is microcitosys. Recently, iron deficiency anemia has been associated to thrombotic phenomenon has raised interest in the studying of erytrocyte deformability, in order to understand such cases.

Three experiments were conducted to study the effect of zinc deficiency on hemoglobin, red cell counts, red cell fragility, storage of erythropoietic minerals in livers and circulating levels of zinc, iron and copper in the rat. In the experiment 1, twenty weanling rats were divided into four groups. Two groups were fed zinc-deficient diet ( < 0.5 ppm); and two groups, zinc-supplemented diet (50 ppm). The rats in the first and third groups were bled three times a week (bled), however the rats in remaining groups were bled once a week (non-bled). All rats received ad libitum food. Mean food intake per day was considerably less for zinc-deficient rats than that for zinc-supplemented rats. Approximately 2 to 3 times as much food was required for each gram of body weight gained by the zinc-deficient group as by the controls. In the experiment 2, fifty-two male weanling rats were divided into six groups; three groups were bled and three non-bled. The 2nd and 5th groups of (zinc-supplemented) rats were pair-fed with the same amount of food as the 1st and 4th zinc-deficient groups ate. The 3rd and 6th groups (control) were fed ad libitum. Hemoglobin levels were lower (P < 0. 05) in bled groups than in the non-bled groups irrespective of zinc treatment in experiment 1 and 2. Within the bled and non-bled groups the zinc-deficient rats consistently exhibited decreased hemoglobin values as compared to zinc-supplemented rats. All zinc-deficient rats had a significantly lower (P < 0. 05) liver zinc content than did the zinc-supplemented ones. There was no difference in iron as well as in copper content of the livers among the groups of experiment 1. However, in experiment 2, copper content per liver was significantly less (P < 0.05) in zinc-deficient rats than in t he zinc-supplemented rats. Serum zinc values were lower in zinc deficient than in control rats. No differences in the concentration of copper in the serum were observed. Red~blood-cell membrane fragility in experiment 2 indicated that there were no differences in percent hemolysis among bled groups. However, percent hemolysis was significantly lower (P < 0.05) in zinc-deficient non-bled groups which clearly indicated that treatment did affect hemolysis. Among other possibilities this might be attributed to more rapid turn-over of red-blood-cells in zinc-deficient rats. In the experiment 3, twenty-four male weanling rats were divided into three groups; one group of zinc-supplemented rats was pair-fed to the zinc-deficient group; and remaining control group, fed ad libitum. All the rats were bled once a week. After 28 days of feeding the rats were injected with 2-c14-glycine (5μc/100 g body weight). It was observed that incorporation of glycine into hemoglobin in the zinc deficient rats was significantly (P < 0.05) more than in the zinc-supplemented rats. With the limited data it is difficult to draw a definite conclusion. Perhaps, it may be true that the red-blood-cells of zinc-deficient rats might have a short life span resulting in the new red cell formation.

Introdução: A pré-eclâmpsia é uma das principais causas de mortalidade materna e perinatal em todo o mundo. A relação entre a amplitude de distribuição das hemácias (Red Cell Distribution Width - RDW) e hipertensão arterial já está bem documentada, porém há uma escassez de dados relacionando RDW com pré-eclâmpsia. Material e métodos: Estudo observacional analítico retrospectivo, realizado no período de 2014 e 2015, composto por 108 participantes no grupo de estudo (50 pré-eclâmpsia leve e 58 pré-eclâmpsia grave) e 101 participantes no grupo controle. A hemoglobina, RDW, plaquetas e outros índices hematológicos foram medidos como parte do hemograma automatizado. Resultados: Não houve diferença no RDW entre as gestantes do grupo controle e grupo com pré-eclâmpsia leve (14,68 ± 1,64 vs. 14,22 ± 1,87; p=0,385), grupo controle e grupo com pré-eclâmpsia grave (14,68 ± 1,64 vs. 14,24 ± 1,78; p=0,386) e grupo controle e grupo com pré-eclâmpsia (14,68 ± 1,64 vs. 14,23 ± 1,81; p=0,063). Conclusão: Os níveis de RDW sérico materno não estão associados com a presença da pré-eclâmpsia e os graus de gravidade da doença. / Introduction: Preeclampsia is one of the main causes of the maternal and perinatal mortality all over the world. The relationship between the amplitude of red cells distribution (Red Cell Distribution Width – RDW) and arterial hypertension is already well documented, though there is a shortage of data relating RDW with preeclampsia. Methods: Analytical observational study conducted in the years 2014 and 2015, composed by 108 patients in the study group (50 mild preeclampsia and 58 severe preeclampsia) and 101 patients in the control group. The hemoglobin, RDW, platelets and other hematological indices were measured as part of the automated hemogram. Results: There was no difference in RDW between the pregnant women in the control group and the ones in the mild preeclampsia group (14,68 ± 1,64 vs. 14,22 ± 1,87), the control group and the severe preeclampsia group (14,68 ± 1,64 vs. 14,24 ± 1,78) and the control group and preeclampsia group (14,68 ± 1,64 vs. 14,23 ± 1,81). Conclusion: The levels of maternal serum RDW are not associated with the presence of preeclampsia and the levels of its severity. / Dissertação (Mestrado)

Introduction: Acute myeloid leukemia (AML) is a highly heterogeneous disease which renders risk stratification at diagnosis of high importance to personalize therapy. Allogeneic hematopoietic stem cell transplantation (HSCT) offers the highest chance for sustained remission in most AML patients, but usually comes at the risk of a significant treatment-related mortality. The red cell distribution width (RDW) is an universally accessible parameter that identifies individuals with a higher mortality in many diseases, including some hematological entities. However, the impact of diagnostic RDW levels in AML – especially in the context of a HSCT consolidation - has not been evaluated so far. Purpose: To evaluate the prognostic impact of RDW levels at AML diagnosis. Methods: A total of 294 newly diagnosed AML patients (median age 60.6, range 14.3-76.5 years), with available diagnostic RDW levels were retrospectively included in this analysis. All patients received a consolidation therapy with an allogeneic HSCT in curative intention between August 2007 and December 2020 at the University Medical Center Leipzig. The RDW was measured in all patients at AML diagnosis before the start of cytoreductive therapies. Results: RDW levels at diagnosis were highly variable (median 16.6%, range 12%-30.6%) and above the upper level of normal (>15%) in 73% of the analyzed AML patients. Patients with RDW levels above 15% did not have worse outcomes compared to patients with low diagnostic RDW levels. However, when the cohort was dichotomized according to a receiver operating characteristic (ROC)-based optimal cut-point (20.7%), patients with high RDW levels had a significantly higher non-relapse mortality (NRM), shorter overall survival and a trend for shorter event-free survival, while the risk of relapse or disease progression was similar in both groups. In multivariate analyses, the RDW remained an independent prognostic factor for higher NRM after adjustment for the body mass index at diagnosis. Patients with a higher RDW were more likely to harbor a secondary AML, as well as to harbor secondary AML-associated gene mutations (i.e. JAK2, ASXL1, or spliceosome mutations, especially SRSF2). Conclusion: High RDW levels at diagnosis represent an independent risk marker for a higher mortality following allogeneic HSCT. When confirmed in prospective clinical trials, the RDW might help to personalize AML consolidation therapy including conditioning regimens before allogeneic HSCT.:1. Bibliographische Beschreibung 2. Abkürzungsverzeichnis 3. Einführung / Introduction 3.1. Acute Myeloid Leukemia 3.1.1. Definition 3.1.2. Epidemiology and etiology 3.1.3. Clinical presentation 3.1.4. Diagnosis of AML 3.1.4.1. Morphology 3.1.4.2. Immunophenotyping 3.1.4.3. Cytogenetic and molecular analyses 3.1.5. AML classification according to WHO classification 3.1.6. Prognostic factors in AML 3.1.6.1. Patient-related risk factors 3.1.6.2. Genetic risk factors 3.1.6.3. Measurable residual disease 3.1.7. Treatment of AML 3.1.7.1. Induction therapy in curative intention 3.1.7.2. Consolidation therapies 3.1.7.3. Palliative treatment approaches 3.1.7.4. New substances 3.2. Allogeneic HSCT 3.2.1. Principles of allogeneic HSCT 3.2.2. Conditioning regimens 3.3. Red cell distribution width 4. Aufgabenstellung / Objectives 5. Materialien und Methoden / Materials and Methods 5.1. Patients and treatments 5.1.1. Treatment protocols 5.1.2. Allogeneic HSCT and immunosuppression 5.1.3. Assessment of GvHD 5.2. Disease characterization 5.2.1. Evaluation at AML diagnosis 5.2.1.1. Morphology 5.2.1.2. Flow cytometry 5.2.1.3. Genetic analyses 5.2.1.4. Evaluation of RDW levels 5.2.2. Evaluation at HSCT 5.2.2.1. Definition of remission status at HSCT 5.2.2.2. Evaluation of measurable residual disease at HSCT 5.3. Statistical Analyses 5.3.1. Associations 5.3.2. Clinical endpoints 5.3.3. Definition of an optimal cut-point for RDW levels 5.3.4. Multivariate analyses 6. Ergebnisse / Results 6.1. Overall outcomes of the patient cohort 6.2. RDW levels at AML diagnosis regarded as continous parameter 6.3. The role of RDW levels at diagnosis as a predictor for outcomes after allogeneic HSCT 6.4. Associations of RDW levels at diagnosis 7. Diskussion / Discussion 8. Zusammenfassung / Summary 9. Literaturverzeichnis / References 10. Erklärung über die eigenständige Abfassung der Arbeit 11. Curriculum Vitae 12. Komplette Publikationsliste (Peer-reviewed) 13. Danksagung

Mon travail de thèse est fondé sur la recherche d’une meilleure compréhension de la distribution géographique des polymorphismes érythrocytaires: antigènes de surface des systèmes de groupes sanguins érythrocytaires (GSE) et glucose-6-phosphate déshydrogénase (G6PD) intracellulaire. L’analyse sur 75 populations d’Asie des fréquences de l'allèle DI*01 du système de GSE Diego, des haplotypes C2-M217 et C2-M401 du chromosome Y, des coordonnées géographiques et des langues, a pu montrer la corrélation entre ces marqueurs. La répartition de DI*01 semble suivre les conquêtes mongoles, avec une expansion radiale depuis la Mongolie, porté par les nomades de langue Altaïque présentant C2-M217 et C2-M401. L'étude du gène G6PD chez 80 individus de Guyane Française de la communauté des Noirs Marrons originaire d’Afrique sub-Saharienne, aborde les relations santé-environnement. Les mutations du déficit en G6PD caractéristiques des variants sub-Sahariens ont été retrouvées chez une personne sur huit. La répartition du déficit était inconnue en Guyane Française et est encore mal connue en Amérique Latine et dans les Caraïbes, où sévit encore Plasmodium vivax dont le traitement nécessite l’utilisation de la primaquine pouvant entraîner une hémolyse sévère chez les individus G6PD-déficients. Mon troisième objectif a été de mettre en évidence l’influence de différents facteurs sur la répartition des polymorphismes de 10 systèmes de GSE étudiant 343 populations. Par des modélisations, les fréquences alléliques ont été confrontées aux données environnementales et culturelles. Enfin, une étude a été réalisée sur le système Duffy avec des analyses de détection de la sélection sur données SNP. / My Ph.D. work is based on the search for a better understanding of the geographical distribution of red blood cell polymorphisms: the surface antigens of red cell blood group systems (BGS) and the intracellular glucose 6-phosphate dehydrogenase (G6PD). The analysis on 75 Eurasian populations of frequencies of the DI*01 allele coding for Diego a antigen of Diego BGS, the C2-M217 and C2-M401 haplotypes of the Y chromosome, geographic coordinates and languages, has shown a correlation between these markers. The DI*01 distribution seems to follow the Mongol conquests, carried by the Altaic-speaking nomads possessing the C2-M217 and C2-M401 haplotypes with a radial expansion from Mongolia. The study of the G6PD gene in 80 individuals from French Guiana of the Noir Marron community originating from sub-Saharan Africa, addresses health-environment relations. Characteristic mutations of sub-Saharan variants of G6PD deficiency have occurred in one in eight people. The G6PD deficiency distribution was previously unknown in French Guiana and is still poorly known in Latin America and the Caribbean, where Plasmodium vivax still cracks down. Its treatment requires the use of primaquine which may cause severe haemolysis in G6PD-deficient individuals. My third objective was to highlight the influence of different factors on the distribution of polymorphisms of 10 BGS studying 343 populations. Through model adjustments, allelic frequencies have been confronted to environmental and cultural data. Finally, a study has been also conducted on the Duffy BGS by analyses of detection of natural selection on SNP data.

No description available.

The discovery and taxonomic history of fluttering shearwater (Puffinus gavia (Forster) and Hutton's shearwater (Puffinus huttoni Mathews) are reviewed. Taxonomic theory, where appropriate to this thesis, is discussed. The external morphology of P. gavia and P. huttoni is compared. No single external measurement or plumage character separates more than 60% of birds examined. The best system of identification is to compare the ratio of different body parts within an individual bird. The distribution of P. gavia and P. huttoni is compared. Hutton's shearwater feeds further out to sea and it is believed to be a migrant species wintering in north west Australian waters. The fluttering shearwater is believed to be a semi-migrant species with only the juveniles spending time in south east Australia. The red cell enzymes of P. gavia, P. huttoni and P. griseus are compared. There are differences in two esterase loci between gavia and huttoni, while P. griseus is more distantly related. Nei's genetic identity values are calculated. The systematic value of electrophoretic data is discussed. The relationship of an undescribed subfossil shearwater to P. gavia and P. huttoni is discussed. An outgroup analysis to other shearwater species is carried out according to phylogenetic (cladistic) theory. The subfossil shearwater is most closely related to the fluttering shearwater, and these two form a sister group to Hutton's shearwater. These three species are a sister group of P. opisthomelas. The relationship between the many P. assimilis subspecies, the black-backed Manx shearwaters, and the gavia, huttoni and opisthomelas group was not resolved. Puffinus nativitatis is more closely related to the Manx and the little shearwaters than to the P. griseus, P. tenuirostris group.