One in six couples wishing to start a family are infertile. The many causes of infertility include genetic defects that can be single gene, multifactorial or chromosomal (including Y deletions, karyotype abnormalities and gamete aneuploidy). This thesis is concerned with the association between infertility and increased sperm aneuploidy. Specific questions are: should males be screened for sperm aneuploidy before intracytoplasmic sperm injection (ICSI)? Is there a relationship between individual semen parameters and sperm aneuploidy for specific chromosome pairs? What is the role of genome organisation in male gametes and its association with infertility? Whether use of alternative therapy (in this case, traditional Chinese Medicine (TCM)) can be used to improve sperm disomy levels. Statistical analysis of questionnaire data revealed that infertility specialists believed there to be merit in screening sperm aneuploidy levels before ICSI. Evidence is presented for possible chromosome-specific and semen parameter specific mechanisms for sperm aneuploidy as is evidence of genome organisation that may be perturbed in infertile males. Finally, in six males studied, sperm aneuploidy levels improved significantly coincident with TCM. Closer investigation of the biological activity of individual therapeutic herbs and treatment cocktails revealed strong anti-oestrogenic and anti-oxidant properties. This suggests a possible mechanism of action of the herbs and provides the basis from which future placebo controlled clinical trials might continue. Possible criticisms of the work presented here include the unavailability of blood samples from many of the patients (thus preventing karyotype analysis) and the absence of a second control group in our studies on semen parameters. Nevertheless significant steps have been made towards establishing the need for, and the implementation of, a pre-ICSI screening test. Moreover progress has been made towards further understanding the aetiology of sperm aneuploidy and towards the implementation of a new treatment that may, ultimately, augment, or even replace ICSI.

Orientador: Andréa Trevas Maciel-Guerra / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-24T13:17:10Z (GMT). No. of bitstreams: 1 Carvalho_AnneliseBarretode_D.pdf: 1678960 bytes, checksum: 38e5284e80abdc6135eaf891cd8d4e9a (MD5) Previous issue date: 2014 / Resumo: A síndrome de Turner (ST) tem como sinais mais frequentes baixa estatura e disgenesia gonadal; são também encontrados dismorfismos, malformações e afecções adquiridas. O fenótipo é muito variável, dificultando o estabelecimento da suspeita clínica e o diagnóstico precoce. O objetivo deste estudo foi identificar os fatores que discriminam pacientes com ST daqueles sem essa síndrome, a fim de auxiliar os médicos, particularmente pediatras, a levantar precocemente essa hipótese e solicitar o exame do cariótipo. A amostra incluiu 516 pacientes do sexo feminino com essa suspeita clínica (por baixa estatura e(ou) hipogonadismo e(ou) dismorfismos característicos) encaminhadas a serviço especializado em distúrbios da diferenciação do sexo na Universidade Estadual de Campinas, entre janeiro de 1989 e fevereiro de 2012. Foi realizado um estudo descritivo de corte transversal, com a comparação entre as pacientes com e sem ST em relação a dados de história clínica e exame físico por meio do Teste do Qui-Quadrado, Teste T de Student e análises de regressão logística univariada e múltipla. Em 186 casos (36%), a ST foi confirmada pelo cariótipo, com predominância de anomalias estruturais dos cromossomos sexuais (41,9%). Nos casos de ST, o peso ao nascimento (p= 0,024) e a estatura ao diagnóstico em escore z (p<0,001) foram menores, e o índice de massa corpórea (p<0,001), maior. Entre as pacientes de mais de 13 anos e aquelas de mais de 16 anos, foram encontrados com maior frequência atraso puberal (p<0,001) e amenorreia primária (p=0,003), respectivamente, no grupo de ST. Na análise univariada, 19 dos 26 sinais dismórficos avaliados no exame físico foram significativamente mais frequentes na ST; na multivariada, o conjunto de variáveis que permitiu discriminar os grupos com e sem ST na amostra foi, em ordem decrescente: linfedema residual de membros, pescoço alado, cúbito valgo, unhas hiperconvexas, tórax alargado, anomalias de mamilos, nevos pigmentados, hipoplasia de metacarpos, maior peso e menor estatura. A investigação de ST deve ser realizada não apenas na presença de dismorfismos típicos, mas também em fenótipos menos evidentes / Abstract: In Turner syndrome (TS) the most frequent features are short stature and gonadal dysgenesis; there may also be dysmorphic signs, congenital malformations and acquired diseases. The phenotype is highly variable, which makes it difficult to establish the clinical suspicion and to achieve early diagnosis. The aim of this study was to identify factors that discriminate patients with TS from those without this syndrome in order to help physicians, particularly pediatricians, to raise this hypothesis and request a karyotype. The sample comprised 516 female patients with this clinical suspicion (with short stature and(or) hypogonadism and(or) typical dysmorphisms) which were referred to a specialized service for disorders of sex development at State University of Campinas from January 1989 to February 2012. A descriptive transversal study was conducted, with comparison between patients with and without TS regarding clinical history and physical examination by qui-square test, t test and univariate and multiple logistic regression analyses. In 186 cases (36%) TS was confirmed by karyotyping, with predominance of structural sex chromosome abnormalities (41.9%). Patients with TS had lower birth weight (p= 0.024), lower height z-score (p<0.001) and higher body mass index (p<0.001). Among patients aged more than 13 years and those aged more than 16 years there were more frequently pubertal delay (p<0.001) and primary amenorrhea (p= 0.003), respectively, in the group of TS. In univariate analysis, 19 out of the 26 dysmorphic signs were significantly nore frequent in TS patients; in multivariate analysis, the set of variables that discriminated between patients with and without TS were, in descending order: residual lymphedema in limbs, webbed neck, cubitus valgus, hyperconvex nails, broad chest, nipple anomalies, pigmented nevi, hypoplastic metacarpals, hipoplasia de metacarpos, higher weight and lower height. Investigation of TS should be performed not only in the presence of typical dysmorphisms but also in less striking phenotypes / Doutorado / Pediatria / Doutora em Ciências

Comparative cytogenetics of the bed bug Cimex lectularius (Heteroptera: Cimicidae) The human bed bug Cimex lectularius has started enormous spreading to all developed countries of temperate climate zone during the last ten years. Bed bug was almost eradicated by a mass use of DDT in these areas until the 70's in the 20th century. This temporal haematophagous ectoparasite occupies particularly human dwellings and bat roosts. Cimex lectularius shows unusual combination of cytogenetic characteristics, general for all Heteroptera, however, not usual for other organisms. The chromosomes are holokinetic, with completely achiasmatic meiosis and inverted meiosis of the sex chromosomes. Especialy remarkable feature is intraspecific variation of the number of the X chromosomes. The variable number of chromosomes of 43 populations of Cimex lectularius from the Czech Republic and 27 populations from other European countries was studied in the present study. The 10 variants of karyotype were found out by using the "hotplate spreading" method and the standard Giemsa staining. There were male karyotypes with 2n = 29, 30, 31, 32, 33, 34, 35, 37, 42 and 47 chromosomes and two females with peculiar odd number of sex chromosomes X, 2n = 33 and 43, not complementary with any male. A stable number of 2n = 26 autosomes...

Collectively, populations of Acanthocyclops vernalis, a species complex of freshwater copepods, are remarkably similar as to morphology and DNA content, despite variability in chromosome number. Reproductive isolation had been reported among some populations, but with each new investigation the species boundaries and factors that may influence them appeared less clear. To clarify the pattern of biological species within this group of populations, we adopted a comprehensive approach and examined patterns of reproductive isolation in populations for which morphology, chromosome number, DNA content, and 18S rDNA sequences are known. In this study we established nine isofemale lines from four sites in Wisconsin and performed 266 crosses. Crosses within and among these lines were used to relate the degree of reproductive isolation to chromosome differences and to construct a model to explain the origin and maintenance of chromosome number variability. Different gametic and somatic chromosome numbers were observed among specimens within some isofemale lines. In a few cases, gametes with different haploid numbers were produced by a single female. Matings within isofemale lines always produced at least some reproductively successful replicate crosses (produced viable, fertile offspring). Crosses between lines from the same site showed reduced success relative to within-line crosses. Crosses between populations from distant sites showed limited genetic compatibility, producing viable, fertile F1 offspring but infertile F2 adults. One cross between lines with different chromosome numbers (one with 2n = 8 and one with 2n = 10) produced fertile viable offspring, which reproduced for at least 60 generations. These hybrids had either eight or nine chromosomes in the third generation of inbreeding, and eight chromosomes after 20 generations. These hybrids also had reduced nuclear DNA contents at the third generation, a level that persisted through the 20th generation. Successful backcrosses between some hybrids and their parental lines further demonstrated the potential for genetic compatibility among forms with different chromosome numbers. We propose a model in which alterations due to Robertsonian fusions, translocations, and/or loss of chromosomal fragments generate heritable variation, only some of which leads to reproductive isolation. Hence, some of the criteria traditionally used to recognize species boundaries in animals (morphology, DNA content, chromosome number) may not apply to this species complex.

Heterochromatin variants of human chromosome 9 belong to the most common variabilities of human karyotype. The variability involves the large block of constitutive heterochromatin in the pericentric region of chromosome 9, which is composed of various types of repetitive DNA sequences. Those variants can be studied from population epidemiologic, molecular cytogenetic and clinical genetic point of view. We have performed a broad epidemiologic study of the incidence of pericentric inversion of chromosome 9 (inv(9)) and other variants of chromosome 9 in 6 different laboratory cohorts, which included the evaluation of more than 26.000 of cytogenetic reports, the study we published is currently the largest in the world. We expressed the overall incidence of inv(9) to be 1.6% and the total incidence of variants of chromosome 9 to be 3.3-3.9%. Inv(9) was more common in females, however the difference was not statistically significant. Molecular cytogenetic part of the project was based on our own diagnostic approach, which involved the combination of three different commercial FISH probes. Combination of those probes allowed us to differentiate particular subvariants of chromosome 9, which cannot be analyzed only by using G- or C-banding. Using our method, we tested 49 carriers of chromosome 9...

Par une stratégie de dépistage combinant le caryotype et l’hybridation in situ en fluorescence (FISH), une insertion (X;6) présente chez des jumelles avec une leucémie myéloïde aiguë (LMA) et une translocation (12;13) dans deux cas de LMA et un cas de leucémie lymphoblastique aiguë (LLA) ont été mis en évidence. L’insertion (X;6) n’est pas rapportée et serait un variant de la translocation (X;6) rapportée dans 4 cas de LMA, dont un associe un gène de fusion MYB-GATA1. Nous avons mis en évidence la dérégulation de l’expression de ces gènes dans le cas d’insertion sans la présence de fusion MYB-GATA1. De plus, dans le premier cas de translocation (12;13) identifié, ETV6 serait fusionné à CDX2 ou FLT3. Le deuxième cas associe la délétion des gènes miR-15a et miR-16-1 à une fusion d’ETV6 et le troisième cas impliquerait une fusion ETV6- FOXO1. / By a screening strategy combining standard cytogenetics and fluorescent in situ hybridization (FISH), an insertion (X;6) in twins with acute myeloid leukemia (AML) and a translocation (12;13) in 2 cases of AML and a case of acute lymphoblastic leukemia (ALL) have been identified. Insertion (X;6) is not reported and could be a variant of translocation (X;6) described in 4 cases of AML, one of which is associated with a MYB-GATA1 fusion gene. We have identified the disruption of MYB and GATA1 in the insertion but no MYB-GATA1 fusion seems to be present. Other mechanisms could be in play for the disruption of these genes’ expression. Moreover, in the first case of translocation (12;13) identified, ETV6 is fused to either CDX2 or FLT3. The second case associates the deletion of miR-15a and miR-16-1 genes to an ETV6 fusion. In the third case, an ETV6- FOXO1 fusion seems to be involved.

Many primate populations face the threat of extinction due to habitat loss, intensive agriculture, hunting for meat, the pet trade and/or use in traditional medicines. An alternative approach to in situ conservation includes gene banking and the use of assisted reproductive technologies (ART), such as oocyte in vitro maturation (IVM) and in vitro fertilization (IVF). Although many of these 'high-tech' solutions have not yet been proven viable for pragmatic wildlife conservation, basic research and development of these emerging tools can provide necessary information needed to optimize these techniques and institute ART as a routine practice in conservation efforts. A severely limiting factor in the successful application of ARTs is the availability of mature developmentally competent oocytes. Oocyte maturation involves many nuclear and cytoplasmic factors, which can be affected by maturation conditions and female age. In vitro maturation does not have the same success rate across species studied. In primates especially, IVM oocytes exhibit reduced developmental capacity upon fertilization when compared to in vivo matured (IVO) oocytes. This study aimed to investigate possible causes of reduced developmental capacity of primate IVM oocytes using the rhesus macaque (Macaca mulatta) as a model. Research efforts included investigation of ovarian senescence, oocyte karyotype and spindle morphology, and establishment of an optimal sperm cryopreservation protocol for use in IVF. Histological examination of the rhesus ovary demonstrated an age-related pattern of follicle depletion similar to that described in the human ovary. Oocyte karyotype analysis revealed a significant effect of IVM on the frequency of hyperhaploidy. In addition, immunostaining and confocal microscopy demonstrated a significant increase of anomalous chromosome congression on the oocyte metaphase II spindle equator in relation to IVM and donor female age. These results indicate that IVM can produce serious, if not lethal consequences for embryo development. This study presents baseline data on ovarian aging in the rhesus macaque and aspects of nuclear maturation during macaque IVM that may contribute to the design of primate oocyte recovery plans. Implementation of either of two sperm cryopreservation methods originally developed for rhesus and vervet monkeys will aid future investigation of the developmental capacity of IVM oocytes.

Com o intuito de contribuir para a compreensão da evolução cariotípica em mamíferos, realizamos estudos comparativos, utilizando a pintura cromossômica, em dois grupos basais de mamíferos, as preguiças e os marsupiais. Realizamos comparações entre os cromossomos humanos e os cromossomos das preguiças de três dedos Bradypus torquatus e Bradypus variegatus, estabelecendo as homologias. A análise conjunta de nossos dados e daqueles da literatura sobre pintura cromossômica em outras espécies de Xenarthra permitiu identificar ou confirmar sinapomorfias cromossômicas dos grupos assim como características ancestrais. Também realizamos comparações entre os cromossomos X das duas espécies de preguiça e entre os cromossomos X dos marsupiais americanos Marmosops incanus e Metachirus nudicaudatus. Os principais resultados e conclusões estão resumidos a seguir. 1. Os cariótipos de B. torquatus e B. variegatus são semelhantes quanto à correspondência com os cromossomos humanos. As duas espécies apresentaram em comum (a) a presença das associações dos cromossomos humanos (HSA) 4/8, 7/10, 7/16, 12/22, 14/15 e 17/19, (b) a conservação de HSA 5, 6, 9, 11, 13, 14, 15, 17, 18, 20 e X, (c) dois pares compartilhando homologia com HSA 2, 7, 10, 12, 19 e 22, (d) três pares, com segmentos homólogos a HSA 8 e (e) a ausência da associação ancestral de Eutheria HSA 16/19. 2. O cariótipo de B. variegatus (2n=54) é mais rearranjado em relação ao humano do que o de B. torquatus (2n=50), principalmente devido a fissões de cromossomos ancestrais, que levaram ao maior número diplóide dessa espécie. 3. Reunindo os dados para as preguiças B. variegatus e B. torquatus aos das demais espécies de Xenarthra que tiveram estabelecidas as correlações entre seus cromossomos e os cromossomos humanos, confirmamos como características presentes em todas as espécies dessa supraordem (a) a conservação de HSA 9, 13, 17, 18, 20 e X, (b) a presença de dois pares cromossômicos compartilhando homologia com HSA 19 e 22 e (c) a presença das associações HSA 4/8, 7/16, 12/22 e 14/15. 4. Confirmamos a associação HSA 7/10 e a divisão de HSA 8 em três blocos como assinaturas cromossômicas da supraordem Xenarthra, o que concorda com a monofilia do grupo. 5. Mostramos que a associação HSA 17/19, presente nos cariótipos de B. variegatus, B. torquatus e B. tridactylus, parece ser assinatura cromossômica do gênero Bradypus, apoiando a monofilia do grupo. 6. Mostramos que a associação HSA 12/22/16 parece ser uma sinapomorfia cromossômica, unindo as espécies B. variegatus e B. tridactylus. 7. Considerando a correspondência com os cromossomos humanos, verificamos que os cariótipos de B. variegatus e B. tridactylus são os mais semelhantes, no gênero Bradypus. 8. A análise das correspondências entre as sequências dos cromossomos humanos e as sequências dos cromossomos de grupos externos de mamíferos placentários (marsupial e galinha) disponíveis no banco de dados Ensembl, mostrou que a associação HSA 7/10 presente na supraordem Xenarthra também ocorre nesses grupos externos. Confirmando-se a homologia dessa associação entre os grupos, ela deveria ser classificada como ancestral de Eutheria, apoiando a posição basal dos Xenarthra na árvore filogenética dos mamíferos placentários. 9. Nossas análises comparativas permitiram propor um cariótipo ancestral de Xenarthra com número diplóide de 48 cromossomos, incluindo (a) as associações HSA 3/21, 4/8, 7/10, 7/16, 12/22 (2x), 14/15 e 16/19, (b) a conservação dos cromossomos HSA 1, 3, 4, 5, 6, 9, 11, 13, 14, 15, 17, 18, 20, 21 e X, (c) dois pares cromossômicos com homologia a HSA 2, 7, 10, 12, 16, 19 e 22 e (d) três pares com homologia a HSA 8. 10. Entre os Xenarthra, B. torquatus e C. hoffmanni, com os menores números diplóides da supraordem, apresentam os cariótipos mais conservados em relação ao cariótipo que propusemos como ancestral de Xenarthra e também em relação ao mais recente cariótipo proposto como ancestral de Eutheria. 11. A conservação da eucromatina do cromossomo X foi evidenciada nos experimentos de pintura cromossômica interespecífica, entre as preguiças B. torquatus e B. variegatus e entre os marsupiais M. incanus e M. nudicaudatus. Os segmentos heterocromáticos desses cromossomos se mostraram divergentes, não permitindo a hibridação in situ interespecífica. / In an attempt to shed additional light on mammalian karyotype evolution, we studied, by chromosome painting, the chromosomes of species from two mammalian basal groups, sloths and marsupials. We compared human chromosomes with the chromosomes of two species of three-toed sloths, Bradypus torquatus and Bradypus variegatus, establishing homologies. Analyzing together ours and published data on chromosome painting in Xenarthra species allowed us to identify or confirm chromosome synapomorphies and ancestral characteristics. We also used chromosome painting to compare the X chromosomes of Bradypus torquatus and Bradypus variegatus as well as the X chromosomes of two American marsupials, Marmosops incanus and Metachirus nudicaudatus. Our main results and conclusions are summarized below. 1. The karyotypes of both B. torquatus and B. variegatus include (a) the human chromosomes associations HSA 4/8, 7/10, 7/16, 12/22, 14/15 and 17/19, (b) the conservation of HSA 5, 6, 9, 11, 13, 14, 15, 17, 18, 20 and X, (c) the disruption into two blocks of HSA 2, 7, 10, 12, 19 and 22, (d) three pairs sharing homologous segments with HSA 8, and (e) the absence of the ancestral eutherian association HSA 16/19. 2. B. variegatus (2n=54) presents a more rearranged karyotype, in relation to the human karyotype, than B. torquatus (2n=50), in particular due to fissions of ancestral chromosomes, which account for its higher diploid number. 3. Our data on B. variegatus and B. torquatus together with the previously published comparisons between human and Xenarthra chromosomes confirm, as characteristics common to the species of this super-order (a) the conservation of HSA 9, 13, 17, 18, 20 and X, (b) the disruption of HSA 19 and 22 into two blocks, and (c) the presence of the human chromosome associations HSA 4/8, 7/16, 12/22 and 14/15. 4. The human chromosome association HSA 7/10 and the disruption of HAS 8 into three blocks were confirmed as chromosome signatures for the super-order Xenarthra, supporting the monophyly of the group. 76 5. The HSA 17/19 association, which we demonstrated to be shared by B. variegatus, B. torquatus and B. tridactylus karyotypes, appears as a chromosome signature for the genus Bradypus, supporting the monophyly of the group. 6. The HSA 12/22/16 association seems to be a chromosome synapomorphic trait linking the species B. variegatus e B. tridactylus. 7. Take into account the correspondence between human and Bradypus chromosomes we observed that B. variegatus and B. tridactylus karyotypes are the most similar in the genus. 8. Based on the comparison of the human chromosomes sequences to the chromosomes sequences of the chicken and a marsupial species (outgroups to placental mammals), available in Ensembl database, we showed that a HSA 7/10 association, which is present in the super-order Xenarthra, is also present in the karyotype of the two outgroup species. As the homology between this chromosome association in Xenarthra and the outgroups are demonstrated, strong support for the classifications of this association as ancestral to Eutheria and of Xenarthra as a basal group in the eutherian phylogenetic tree will be given. 9. Our comparative analysis allow us to propose an ancestral Xenarthra karyotype with 2n=48, including (a) the human chromosome associations HSA 3/21, 4/8, 7/10, 7/16, 12/22 (2x), 14/15 and 16/19, (b) the conservation of HSA 1, 3, 4, 5, 6, 9, 11, 13, 14, 15, 17, 18, 20, 21 and X, (c) the disruption into two blocks of HSA 2, 7, 10, 12, 16, 19 and 22, and (d) three pairs sharing homologous segments with HSA 8. 10. Among Xenarthra species, B. torquatus and C. hoffmanni, with the lowest diploid number of the super-order, show the most conserved karyotypes in relation to our proposed ancestral Xenarthra karyotype as well as to the most recently proposed ancestral eutherian karyotype. 11. The conservation of the X chromosome euchromatin was demonstrated by interspecific chromosome painting between the sloths, B. torquatus and B. variegatus, and between the marsupials, M. incanus and M. nudicaudatus. The X chromosome heterochromatic segments were shown to be divergent in the extent to prevent in situ hybridization between species.

Submitted by Angela Maria de Oliveira (amolivei@uepg.br) on 2017-10-18T13:46:42Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Barbara de Melo.pdf: 1643171 bytes, checksum: abd718922152e41d663027fa3f15740b (MD5) / Made available in DSpace on 2017-10-18T13:46:42Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Barbara de Melo.pdf: 1643171 bytes, checksum: abd718922152e41d663027fa3f15740b (MD5) Previous issue date: 2013-02-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A subfamília Alticinae é considerada moderna com grande variação de número diplóide e sistema de determinação sexual. Considerando a tribo Systenini apenas seis espécies foram analisadas citogeneticamente e essas espécies possuem grande variação no número cromossômico e no sistema de determinação do sexo. Oedionychina, por outro lado, apresenta espécies com número diploide e sistema de determinação sexual conservados de 2n=22=20+X+y com cromossomos sexuais gigantes e assinápticos. A análise das Regiões Organizadoras de Nucléolo (RONs) tem sido realizada principalmente através de Impregnação pelo Íon Prata, poucas espécies de Alticinae têm sido analisadas através de Hibridação in situ Fluorescente até o presente momento. O mapeamento do gene ribossomal 5S foi analisado apenas em espécies de Scarabaeidae. Com relação ao cístron 45S somente 3 espécies de Alticinae foram descritas. Assim, o objetivo desse trabalho foi caracterizar citogeneticamente seis espécies de Alticinae (Omophoita octoguttata, Omophoita personata, Omophoita magniguttis, Alagoasa coccinelloide, Alagoasa florigera e Systena tenuis), visando estabelecer as diferenças cariotípicas e as estratégias de diferenciação cromossômicas, para compreender as relações evolutivas entre as espécies. A análise citogenética das espécies A. coccinelloide, A. florigera e S. tenuis estão concordantes com os dados da literatura. O número diploide das espécies do gênero Alagoasa é de 2n=22=20+X+y com cromossomos sexuais gigantes e assinápticos que segregam corretamente na anáfase I. S. tenuis apresentou o número diplóide de 2n=32=15II+neoXY. O uso de fluorocromo em S. tenuis mostrou à presença de regiões pericentrométicas ricas em repetições AT. A Hibridação in situ do Genoma confirmou o mecanismo cromossômico neoXY. A realização da FISH com sonda de 18S e 5S evidenciou que em O. octoguttata, O. personata e A. coccinelloide ocorre a presença de um par autossômico portador dos genes ribossomais. O. magniguttis e A. florigera mostraram uma derivação no cariótipo apresentando dois e três pares de cromossomos portadores dos genes ribossomais respectivamente. A dupla FISH mostra que, nas 5 espécies os genes estão colocalizados. S. tenuis evidenciou múltiplos sítios desses genes, e a dupla FISH mostrou os genes estão em cromossomos separados ou colocalizados. A FISH com fibras estendidas mostrou que em todos os casos os genes estão dispostos de forma interespaçada. / The Alticinae subfamily is considered modern with wide variation of the diploid number and sex determination system. Considering the Systenini tribe only 6 species were cytogenetically analyzed and these species exhibit wide variation in the chromosome number and the system of sex determination. Oedionychina, on the other hand, present species with diploid number and sex determination system conserved of 2n=22=20+X+y, with extremely large and asynaptic sex chromosomes. The analysis of Nucleolus Organizer Regions (NORs) has been attempted mainly by impregnation of the Ag-NOR, and a few species of Alticinae have been examined by using Fluorescent in situ Hybridization to date. The mapping of 5S ribosomal gene was analyzed only in Scarabaeidae species. In relation to the 45S cistron only three species of Alticinae have been described. The aim of the present study was characterize cytogenetically 6 species of Alticinae (Omophoita octoguttata, Omophoita personata, Omophoita magniguttis, Alagoasa coccinelloide, Alagoasa florigera e Systena tenuis), in order to determine the karyotype differentiation and the strategies of chromosome differentiation to understand the evolutionary relation among the species. The cytogenetic analysis of A. coccinelloide, A. florigera and S. tenuis are in accordance with the literature data. The diploid number of the Alagoasa species is 2n=22=20+X+y with large sex chromosomes, which are asynaptic although have regular segregation in anaphase I. Systena tenuis presented the diploid number of 2n=32=15II+neoXY. The fluorocrome analysis in S. tenuis showed the presence of pericentromeric region rich in AT repetitions. The Genome In situ Hybridization confirmed the system of sex determination of the type neoXY. Performing FISH with 18S and 5S probe revealed the presence of one autosomal pair carrier the ribosomal genes in O. octoguttata, O. personata and A. coccinelloide. Derivations on karyotype showing two or three pairs of chromosomes carrying of the ribosomal genes have been observed in O. magniguttis and A. florigera, respectively. The double FISH showed that genes are colocalized in the five species. S. tenuis showed multiple sites of these genes, and the double FISH showed the genes are in separate chromosomes or colocalized. The extended fiber FISH showed that in all cases the genes are interspersed arranged.

Made available in DSpace on 2016-08-19T17:47:09Z (GMT). No. of bitstreams: 1 Antonio Alberto Carvalho.pdf: 630872 bytes, checksum: 13ecf29f7245808d06388075ae78be17 (MD5) Previous issue date: 2006-06-16 / In this study the results of the evaluation cytogenetics of 694 patients with suspected of being carriers of chromosomal abnormality, seen in Sarah Network of Hospitals for Rehabilitation - San Luis Unit, in the period from December 1996 to January 2005. / Neste estudo são apresentados os resultados da avaliação citogenética de 694 pacientes, com suspeita de serem portadores de anomalia cromossômica, atendidos na Rede Sarah de Hospitais de Reabilitação - Unidade São Luís, no período de dezembro de 1996 a janeiro de 2005.