Sardinia was an island with a history of a malarious environment until eradication efforts were conducted from 1946 to 1950. While historic documents suggest the disease was introduced from North Africa around 500 BC, no study has been conducted to test for the presence of malaria in prehistoric native populations, such as the Nuragic people of the Bronze Age. However, it has been suggested that aspects of the Nuragic culture, for example the stone structures found throughout the island, are adaptations to a malarious environment. The purpose of this dissertation is to test the hypothesis that malaria was present in prehistoric Sardinia. In addition, the value of applying anthropology, pertaining specifically to prehistoric investigations, to understand and combat malaria is supported. To test for the presence of malaria, multiple lines of evidence were used to analyze human skeletal remains from a Middle Bronze Age tomb. Because malaria does not result in a specific pattern of bony responses that can be identified through a gross analysis of the remains, additional lines of evidence were used. These included an osteological analysis for the possible presence of conditions related to malaria (e.g., inherited hemolytic anemias) and the collection of bone samples to test for ancient malaria DNA, Plasmodium falciparum histidine-rich protein II, and the malarial pigment hemozoin. In addition, a review of the literature pertaining to the ecology and history of Sardinia were used with archaeological data to evaluate if it was possible the malaria parasite was affecting humans on the island during prehistory. While it was interpreted that conditions were favorable for malaria to infect individuals during this time, and possible cultural adaptations were noted, no conclusive evidence was found by analyzing skeletal remains. More work is needed to diagnose malaria better in human remains and understand the health of populations in Sardinia during the Bronze Age. Considering the coevolution of malaria parasites, humans, and mosquitoes is a necessary step in developing methods to combat malaria as the parasite and disease vector become more resistant to medicine and insecticides. In particular, applying anthropological methods and theories shows promise for fighting this disease.

<p>The purpose of this study is to assess the feasibility of a non-invasive, reliable, and cheap method to evaluate iron-overload in beta-thalassemia major patients. The approach taken was through the possibility of in-vivo measurement of iron in the skin using a technique called x-ray fluorescence. It was hoped that the quantification of iron levels in the skin will correlate with those levels in major parenchymal organs, such as the liver and the heart – where most iron deposition occurs in thalassemic patients. Water phantoms were used to produce a calibration line with an R² value of 0.998. Skin, liver and heart tissues from 36 control mice were measured and their iron levels quantified. Iron concentration range in the skin was found to be -2 – 38 ppm with an average of 9.8 ± 1.6 ppm. Significant correlation was found between the iron levels in skin vs. heart (R_s² of 0.382); however, it was not significant in skin vs. liver (R_s² of 0.080). Skin biopsies from various sites of 6 cadavers were investigated in a synchrotron light source facility. Maps of iron, zinc and calcium distribution as a function of skin depth were then constructed. It was found that all three elements were significantly present in the epidermal layer compared to the dermal one. Calcium and zinc were present in the entire epidermis, whereas iron was mainly concentrated at the deepest region of the epidermis. It was also concluded that skin samples from the back, arm and thigh gave the clearest elemental distribution.</p> / Master of Science (MSc)

This thesis investigates the use of x-ray fluorescence (XRF) measurements of skin iron concentration as a non-invasive surrogate marker for total body iron burden. Rats were overloaded with iron dextran via intraperitoneal injection to investigate trends in iron accumulation in skin and organs. The skin, heart, liver, and kidney of the animals were collected and the iron concentrations were measured using the Huber XRF system and the polarized energy dispersive x-ray fluorescence (PEDXRF) system. When measured in the Huber XRF system, a very strong quadratic correlation was found between skin iron concentration and liver iron concentration (R2=0.92). In the same system, skin iron concentration had a moderately strong linear correlation with heart and kidney iron concentration (R2=0.53 and R2=0.65, respectively). Measured in the PEDXRF system, heart and kidney iron concentrations were again linearly correlated with skin iron concentrations (R2=0.34 and R2=0.31, respectively). Liver iron concentration again showed a quadratic correlation with skin iron concentration (R2=0.64). Therefore, it was demonstrated that skin iron concentrations can act as a surrogate marker for organ iron concentrations in rats, especially for the liver. The feasibility of using an Olympus Innov-X Delta Professional Handheld XRF Analyzer, a commercial portable x-ray fluorescence (PXRF) device, as a tool for in vivo skin iron analysis was investigated. The same rat skin samples measured in the previous experiment were measured using the PXRF device and compared with the organ iron concentrations as measured using the Huber XRF system. The heart and kidney showed linear correlations with skin iron concentration (R2=0.45 and R2=0.36, respectively). The liver showed the strongest correlation with a moderately strong quadratic correlation with R2=0.74. It was determined that using Beam 3 in the Soil mode of this device resulted in equivalent and effective dose rates of 230±10mSv/min and 2.3±0.1mSv/min, respectively. Thus, the PXRF device has shown promise as a potential tool for measuring in vivo skin iron levels. / Thesis / Master of Science (MSc)

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

Thesis (Ph. D.)--University of Hong Kong, 1996. / Includes bibliographical references (leaf 370-410).

Thesis (Ph.D.)--University of Alabama at Birmingham, 2009. / Title from PDF title page (viewed on July 13, 2010). Includes bibliographical references.

Made available in DSpace on 2014-06-11T19:26:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-25Bitstream added on 2014-06-13T19:12:53Z : No. of bitstreams: 1 carrocini_gcs_me_sjrp.pdf: 1450758 bytes, checksum: 22d1c253bc1aae8bf01a4211133799fb (MD5) / A hemoglobina Fetal (Hb F), formada por duas cadeias globínicas alfa (α) e duas gama (γ) (α2 γ2), apresenta expressão reduzida em indivíduos adultos, variando de 0% a 1% do total de hemoglobinas. Os níveis aumentados de Hb F são decorrentes de mutações na família β- globina, que originam a Persistência Hereditária de Hemoglobina Fetal (PHHF) e a Delta- Beta Talassemia (δβ-Talassemia). O controle na produção se dá por região reguladora e por regiões não pertencentes à família β-globina, dentre elas 2q16, 6q23, 8q e Xp22.2. Os objetivos do presente trabalho consistiram em verificar a presença e a frequência de duas mutações para a δβ-Talassemia, do polimorfismo XmnI e dos haplótipos β-globina em indivíduos saudáveis com Hb F aumentada do estado de São Paulo. Foram analisadas 60 amostras de sangue periférico de indivíduos adultos, saudáveis e sem queixa de anemia. Foram separadas em dois grupos de estudo, de acordo com os níveis de Hb F em grupo I - 34 amostras com Hb F variando de 2% a 15%; e grupo II - 26 amostras com níveis de Hb F superiores a 15%. Em relação aos polimorfismos analisados, foram encontrados três indivíduos heterozigotos (5%) para a mutação δβ-Talassemia Espanhola, pertencentes ao grupo I, cujos níveis de Hb F estavam dentro da faixa de normalidade. A mutação δβ- Talassemia Siciliana não foi encontrada, fato que requer a investigação de outros polimorfismos relacionados ao aumento de Hb F na vida adulta. A frequência do polimorfismo XmnI foi de 33,3% e os níveis médios de Hb F foram de 15,48 ± 11,69%. A frequência observada em nosso estudo para esse sítio polimórfico é maior do que os achados da literatura para indivíduos saudáveis. Esse polimorfismo foi mais presente em indivíduos com concentrações de Hb F abaixo de 15%. Para quatro das amostras positivas para esse polimorfismo os níveis... / Fetal Hemoglobin (Hb F), formed by two alpha globin chains (α) and two gamma chains (γ) (α2 γ 2 ), has reduced expression in adults, ranging from 0% to 1% of total hemoglobin. Increased levels of Hb F are due to mutations in β-globin family, which cause the Hereditary Persistence of Fetal Hemoglobin (HPFH) and Delta-Beta Thalassemia (δβ-Thalassemia). The control of the production takes place by the regulatory region and regions outside the β-globin family, among them 2q16, 6q23, 8q and Xp22.2. The aims of this study were to verify the presence and frequency of two mutations for δβ-thalassemia, of the XmnI polymorphism and β-globin haplotypes in healthy individuals with increased Hb F of the state of Sao Paulo. We analyzed 60 samples of peripheral blood of adults, healthy and without complaints of anemia. The samples were separated into two groups, according to the Hb F levels in group I - 34 samples with Hb F ranging from 2% to 15% and group II - 26 samples with Hb F levels above 15%. In relation to the polymorphisms examined, we found three heterozygous individuals (5%) for Spanish δβ-thalassemia, belonging to group I, whose Hb F levels were within the normal range. The Sicilian δβ-thalassemia mutation was not found, fact that requires the study of other polymorphisms related to the increase of Hb F in adult life. The frequency of XmnI polymorphism was 33.3% and the mean Hb F levels were 15.48 ± 11.69%. The frequency observed in our study to this polymorphic site is higher than the findings in the literature for healthy subjects. This polymorphism was more prevalent in individuals with Hb F concentrations below 15%. For four samples positive for this polymorphism, the Hb F levels were explained by the presence of HPFH and Spanish δβ-Thalassemia mutations, so that the presence of XmnI polymorphic site was not determinative to influence the γ-globin genes expression... (Complete abstract click electronic access below)

Made available in DSpace on 2014-12-17T14:16:25Z (GMT). No. of bitstreams: 1 ZamaMLS_DISSERT.pdf: 2983727 bytes, checksum: 7afd93b3940a02c3ef820577c21fb808 (MD5) Previous issue date: 2010-02-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Beta thalassemia arises as a consequence of the reduction (?+, ?++, ?silent) or absence (?0) of beta globin chain synthesis and results from a number of mechanisms that lead to genetic defects. The inheritance of beta thalassemia is characterized by the existence of heterozygous individuals, compound heterozygotes, homozygotes and those with coinheritance of beta thalassemia allele and other thalassemias and/or hemoglobin variants. The aim of this study was to perform molecular and laboratory characterization of beta thalassemia in heterozygous and homozygous individuals and in those with coinheritance of S beta thalassemia. A total of 48 individuals were included (35 heterozygotes, 4 homozygotes and 9 S beta thalessemia carriers) referred to the Integrated Laboratory of Clinical Analyses of the Federal University of Rio Grande do Norte (UFRN) and the Hematology Ambulatory Facility of the Dalton Barbosa Cunha Hemocenter (Hemonorte Natal, Brazil). Peripheral blood samples form each patient underwent the following laboratory examinations: erythrogram, hemoglobin electrophoresis at alkaline pH, measurements of Hb A2, Fetal Hb and serum ferritin. DNA was extracted using the illustra blood genomicPrep Mini Spin Kit and molecular characterization was performed by the PCR/RFLP technique, which involves digestion with specific restriction enzymes for IVS-1 nt 1 (G?A), IVS-1 nt 6 (T?C) and codon 39 (CAG?TAG) mutations. Of the 35 heterozygotes, 37.1% showed IVS-1 nt 6 mutation, 42.9% IVS-1 nt 1 and 20% were carriers of other mutations not identified by the technique used. The four homozygous patients presented with the IVS-1 nt 6 mutation, while 66.7% of the individuals with S beta thalassemia had the IVS-1 nt 1 mutation. Codon 39 was not detected in any of the patients investigated. Of the thallasemic alleles found, 40.4% were IVS- 1 nt 1, 40.4% IVS-1 nt 6 and 19.2% were not identified. Laboratory data showed that the heterozygotes exhibited microcytosis and hypochromia, evidenced by MCV ranging from 57 to 75fL and MCH from 15.9 to 23.6 pg. Hemoglobin A2 varied between 3.7 and 7.2%. The homogygotes also showed reduced MCV and MCH and elevated HbA2.. Comparison of laboratory data between heterozygous individuals with IVS-1 nt 1 and IVS-1 nt 6 mutations showed that heterozygotes for the IVS1-1 mutation had significantly lower mean MCV and MCH (p = 0.023 and 0.007, respectively) and significantly higher hemoglobin A2 (p < 0.001) when compared to heterozygotes for the IVS-1 nt 6 mutation. PCR/RFLP was useful in identifying the presence or absence of IVS-1 nt 6, IVS-1 nt 1 and codon 39 mutations in most of the patients investigated here. This is the first study conducted in the state of Rio Grande do Norte, Brazil aimed at identifying beta thalassemia mutations and represents an important contribution to the knowledge regarding the molecular profile of beta thalassemia in our country / A talassemia beta ocorre devido ? diminui??o (?+, ?++, ?silent) ou aus?ncia (?0) de s?ntese de cadeias beta de globina e ? decorrente de v?rios mecanismos que provocam o defeito gen?tico. A heran?a da talassemia beta ? caracterizada pela exist?ncia de indiv?duos heterozigotos, heterozigoto compostos, homozigotos e portadores de intera??o entre o alelo talass?mico beta e outras talassemias e/ou variantes de hemoglobina. O objetivo principal deste estudo foi realizar a caracteriza??o molecular e laboratorial em indiv?duos heterozigotos e homozigotos da talassemia beta e em portadores da intera??o hemoglobina S/talassemia beta. Foram inclu?dos 48 indiv?duos (35 heterozigotos, 4 homozigotos e 9 com intera??o HbS/talassemia beta) atendidos no Laborat?rio Integrado de An?lises Cl?nicas (UFRN) e no Ambulat?rio de Hematologia do Hemocentro Dalton Barbosa Cunha (Hemonorte Natal/RN). As amostras de sangue perif?rico de cada paciente foram submetidas aos seguintes exames laboratoriais: eritrograma, eletroforese de hemoglobina em pH alcalino, dosagem das hemoglobinas A2 e Fetal, e ferritina s?rica. O DNA foi extra?do utilizando-se o kit blood genomicPrep Mini Spin e a caracteriza??o molecular foi realizada atrav?s da t?cnica PCR/RFLP mediante digest?o com enzimas de restri??o espec?ficas para as muta??es IVS-1 nt 1 (G?A), IVS-1 nt 6 (T?C) e nonsense c?don 39 (CAG?TAG). Dentre os 35 heterozigotos, 37,1% apresentaram a muta??o IVS-1 nt 6, 42,9% a IVS-1 nt 1 e 20% eram portadores de outras muta??es n?o identificadas com a t?cnica utilizada. Os quatro pacientes homozigotos apresentaram a muta??o IVS-1 nt 6, enquanto 66,7% dos indiv?duos com intera??o HbS/talassemia beta tinham a muta??o IVS-1 nt 1. A c?don 39 n?o foi detectada em nenhum dos pacientes investigados. Dentre os alelos talass?micos encontrados, 40,4% eram IVS-1 nt 1, 40,4% eram IVS-1 nt 6 e 19,2% n?o foram identificados. Em rela??o aos dados laboratoriais, os heterozigotos apresentaram microcitose e hipocromia evidenciada pelo VCM variando de 57 a 75fL, e o HCM, entre 15,9 a 23,6 pg. A hemoglobina A2 variou de 3,7 a 7,2%. Os homozigotos tamb?m apresentaram VCM e HCM reduzidos e HbA2 elevada. Ao comparar os dados laboratoriais entre os indiv?duos heterozigotos para as muta??es IVS-1 nt 1 e IVS-1 nt 6 observou-se que os heterozigotos da muta??o IVS1-1 apresentaram valores m?dios de VCM e HCM significativamente menores (p = 0,023 e 0,007, respectivamente) e hemoglobina A2 significativamente mais elevados (p < 0,001) quando comparados aos heterozigotos da muta??o IVS-1 nt 6. A t?cnica de PCR/RFLP se mostrou ?til para a identifica??o da presen?a ou aus?ncia das muta??es IVS-1 nt 6, IVS-1 nt 1 e ?0 c?don 39 na maioria dos pacientes investigados na pesquisa. O presente estudo ? o primeiro trabalho realizado no estado do Rio Grande do Norte para identifica??o das muta??es da talassemia beta, e constitui importante contribui??o para o conhecimento do perfil molecular da talassemia beta em nosso pa?s

Οι αιμοσφαιρινοπάθειες συγκαταλέγονται ανάμεσα στις πιο κοινές μονογονιδιακές διαταραχές παγκοσμίως, συμπεριλαμβανομένης της β-θαλασσαιμίας και της δρεπανοκυτταρικής αναιμίας. Η επανενεργοποίηση των γονιδίων της γ-σφαιρίνης φαίνεται να είναι μια ενδιαφέρουσα θεραπευτική προσέγγιση για τους ασθενείς που πάσχουν από β-τύπου αιμοσφαιρινοπάθειες. Ορισμένες φαρμακευτικές ουσίες έχουν τη δυνατότητα να επάγουν παροδικά την έκφραση των γονιδίων της γ-σφαιρίνης γεγονός που βελτιώνει το φαινότυπο των ασθενών λόγω των υψηλότερων επιπέδων HbF που παρατηρούνται. Η μόνη φαρμακευτική ουσία που έχει εγκριθεί από τον FDA και χρησιμοποιείται ευρύτατα σε ασθενείς που πάσχουν από β-τύπου αιμοσφαιρινοπάθειες και συγκεκριμένα από δρεπανοκυτταρική αναιμία είναι η HU. Παρά το γεγονός ότι στην πλειονότητα των ασθενών παρατηρείται αύξηση της παραγωγής HbF μετά από αγωγή με HU (Steinberg et al. 1997), τα επίπεδα αύξησης διαφέρουν αρκετά μεταξύ των ασθενών που πάσχουν από β-θαλασσαιμία και δρεπανοκυτταρική αναιμία(Patrinos and Grosveld 2008). Σύμφωνα με τους Borg και συνεργάτες, το γονίδιο KLF10 φαίνεται να σχετίζεται με την αύξηση των επιπέδων της HbF και την ανταπόκριση ασθενών με β-τύπου αιμοσφαιρινοπάθειες σε θεραπεία με HU(Borg et al. 2012). Στην παρούσα μελέτη διερευνήθηκε η πιθανή συσχέτιση των SNP rs319133 και rs11552577, που εδράζονται στο γονίδιο KLF10, με αυξημένα επίπεδα HbF και η αξιολόγηση αυτών ως φαρμακογονιδιωματικοί δείκτες, που σχετίζονται με τη μεταβλητότητα των επιπέδων της HbF ως απόκριση στη θεραπεία με HU. Χρησιμοποιήθηκαν ασθενείς που πάσχουν από βαριά β-θαλασσαιμία, ενδιάμεση β-θαλασσαιμία, μη-θαλασσαιμικοί ασθενείς και διπλά ετερόζυγοι ασθενείς για β-θαλασσαιμία και δρεπανοκυτταρική αναιμία. Η μέθοδος γονοτύπησης που χρησιμοποιήθηκε και για τους δύο πολυμορφισμούς ήταν η PCR-RFLP. Η απουσία του πολυμορφισμού rs319133 φαίνεται να σχετίζεται με αυξημένα επίπεδα HbF στους ασθενείς που χαρακτηρίζονται από ενδιάμεση β-θαλασσαιμία συγκριτικά με ασθενείς που πάσχουν από βαριά β-θαλασσαιμία (p=0.04). Επίσης παρατηρείται μια στατιστική τάση συσχέτισης χειρότερης ανταπόκρισης στην αγωγή με HU στους διπλά ετερόζυγους ασθενείς 96 για β-θαλασσαιμία και δρεπανοκυτταρική αναιμία. Για τον δεύτερο πολυμορφισμό που μελετήθηκε δεν παρατηρήθηκαν στατιστικά σημαντικές διαφορές. Τα δεδομένα της παρούσας μελέτης υποδεικνύουν συσχέτιση του γονιδίου KLF10 με αυξημένα επίπεδα HbF. Μελέτες σε πολυπληθέστερες ομάδες θα μπορούσαν να οδηγήσουν στον εντοπισμό και άλλων πολυμορφισμών που σχετίζονται με αυξημένη έκφραση των γονιδίων της γ-σφαιρίνης. / Hemoglobinopathies are amongst the most common single gene disorders worldwide, including the thalassemias and sickle cell disease (SCD). Reactivation of the human γ-globin genes would be a therapeutic intervention for β-type hemoglobinopathies patients. Some drugs and compounds can transiently induce γ-globin gene expression and improve the disease phenotype by augmenting HbF accumulation. Only HU (hydroxyurea) is approved from the FDA and is widely used to treat patients with β-type hemoglobinopathies, in particular sickle cell disease. Although the majority of patients show an increase of HbF production upon HU treatment(Steinberg et al. 1997), the level of HbF increase differs considerably among β-thalassemia and SCD patients(Patrinos and Grosveld 2008). According to Borg et al, KLF10 appears to be significantly associated with high HbF and it may act as an important pharmacogenetic biomarker for β-type hemoglobinopathies patients who are treated with HU(Borg et al. 2012). The aim of our study was to elucidate whether there is an association of the SNPs namely rs3191333 and rs11552577 in KLF10 gene with increased levels of HbF and with response to HU treatment in β-hemoglobinopathies patients. We analyzed samples of β-thalassemia major patients, of β-thalassemia intermedia patients, of healthy (non-thalassemic) donors and samples of SCD/β–thalassemia patients who have been treated with HU. Genotyping was carried out by using PCR-RFLP. The lack of rs3191333 is associated with increased HbF levels in β–thalassemia intermedia compared to β– thalassemia major patients (p=0.04). Also, it is shown a statistical trend of worse response to HU treatment in compound heterozygote β–thalassemia-SCD patients. As far as rs11552577 is concerned no statistically significant differences were observed. Our data show that KLF10 gene is strongly associated with HbF levels. Further analyses could possibly reveal novel variants which are associates with increased expression of γ-globin genes.