"Whole-Exome Sequencing Enables Rapid Determination of Xeroderma Pigmentosum Molecular Etiology"

Xeroderma pigmentosum (XP) is a rare autosomal recessive disorder haracterized by extreme sensitivity to actinic pigmentation changes in the skin and increased incidence of skin cancer. In some cases, patients are affected by neurological alterations. XP is caused by mutations in 8 distinct genes (XPA through XPG and XPV). The XP-V (variant) subtype of the disease results from mutations in a gene (XPV, also named POLH) which encodes for Polg, a member of the Y-DNA polymerase family. Although the presence and severity of skin and neurological dysfunctions differ between XP subtypes, there are overlapping clinical features among subtypes such that the sub-type cannot be deduced from the clinical features. In this study, in order to overcome this drawback, we undertook whole-exome sequencing in two XP sibs and their father. We identified a novel homozygous nonsense mutation (c.897T.G, p.Y299X) in POLH which causes the disease. Our results demonstrate that next generation sequencing is a powerful approach to rapid determination of XP genetic etiology.

Caracterização da resistência do algodoeiro a Ramularia areola e variabilidade molecular do patógeno

This research was conducted with the aim to study the genetic and pathogenic structure of Ramularia areola isolates collected in Brazil and to characterize the resistance response in cotton plants to ramularia spot. The genetic variability of 28 isolates of R. areola was studied using RAPD markers. The pathogenicity evaluation was realized by the inoculation of 6 isolates on cotton varieties Guazuncho-2 (Gossypium hirsutum) and VH8-4602 (Gossypium barbadense). The inheritance of disease resistance was studied using an artificially inoculated population of F2 individuals derived from the intercross of Guazuncho-2 (susceptible variety) end VH8-4602 (resistant variety), and also the parents and F1 individuals. Molecular polymorphism between the G. hisutum varieties DeltaOpal (suscetible) and CNPA CO-11612 (resistant) was estimated by 118 SSR and 24 AFLP markers. The parental genotypes Guazuncho-2 and VH8-4602 were selected for mapping, and then Recombinant Inbred Lines (RIL´s) derived from this crossing were evaluated with SSR 12 markers. The analysis of population structure of R. areola revealed that the three subpopulations were genetically simillar (Gst=0.18), and the isolates from Goiás and Minas Gerais were more similar to each other (0,92). This probability can be related to the relatively high gene flow among the three subpopulations (Nm=2.20). The isolates R. areola 9.1, from Minas Gerais State and 8.1 and 8.3 from Goiás State were the most aggressive ones to the susceptible variety Guazuncho-2. The variety VH8-4602 presented high level of resistance to ramularia spot. No differential interaction was observed between the pathogens and the analyzed varieties, and the resistance was classified as horizontal. The quantification of disease by number of necrotic lesions and number of spores in individual plants of F1 and F2 generations from the crossing between the varieties Guazuncho-2 and VH8-4602 presented continuous distribution, suggesting polygenic resistance. The resistance is probabilly recessive, since necrotic lesions and sporulation were observed on F1 plants. The molecular polymorphism between DeltaOpal e CNPA CO-11612 lineages was low (6%), then would be difficult to accomplish molecular mapping of disease resistance using this intercross. With the genotyping of the RIL s it was verified that 25% of the markers segregated in the proportions proposed by Mendel s Law and 75% of the studied markers presented segregation distortion in favor to the parental G. hirsutum. Both the low genetic variability of the pathogen and the number of resistance genes suggest that durable genetic resitance may be achieved

Análise molecular dos éxons 8 a 11 do gene da p53 em amostras de câncer de colo de útero no Rio Grande do Norte

Mutations on TP53 gene are common in human cancer but not in cervical cancer where they are rarely found and the inactivation and degradation of p53 protein are attributed to the action of E6 viral oncogene from high risk human papillomavirus (HPV). Analysis of cervical cancer cell lines suggests that HPV negative samples shows mutation on TP53, but clinical approaches didn t confirmed this hypothesis. However, in most TP53 mutations studies on cervical cancer, only the exons 5 to 8 were analyzed. Approximately 90% of mutations described are on this region. Recent studies on several cancer suggests that mutation frequency in the other exons must be considered. The aim of this work was to verify whether mutations on coding and non-coding regions occur in cancer tissue from cervical cancer in patients from Rio Grande do Norte using Denaturing Gradient Gel Electrophoresis (DGGE) as screening tool. Exons 8 to 11 were analyzed including some introns from 80 tumor samples and 8 peripheral blood samples from healthy women. DNA were submitted to PCR using primers with GC clamp on the end of one of them. The results were observed for each region after DGGE and silver staining. It was observed no amplified fragment with different migration profile from those obtained from DNA of peripheral blood. These results agree with those from literature where TP53 mutations in cervical cancer have been described in a very low frequency

Relações filogenéticas e filogeografia molecular das espécies de peixes anuais do gênero Simpsonichthys (Cyprinodontiformes: Rivulidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização citogenética e molecular das espécies pintado (Pseudoplatystoma corruscans), cachara (Pseudoplatystoma reticulatum) e seus híbridos utilizados na piscicultura brasileira

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Variação genética para caracteres silviculturais e marcador molecular em uma população base de Eucalyptus camaldulensis Dehnh

Pós-graduação em Agronomia - FEIS

Preservação, caracterização patogênica e molecular de Plasmodiophora brassicae, agente causal da hérnia das crucíferas

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Estrutura molecular e citogenética de cromossomos B em Astyanax paranae (Characiformes, Characidae)

Pós-graduação em Ciências Biológicas (Genética) - IBB

Caracterização molecular de genes do sistema imune de búfalo

Pós-graduação em Genética e Melhoramento Animal - FCAV

Caracterização molecular do gene que codifica a TCTP (Translationally Controlled Tumor Protein) em tomate (Solanum lycopersicum cv. Santa Clara)

The gene encoding TCTP (Translationally Controlled Tumour Protein) is present in all eukaryotes and its product is involved in various cellular processes. Although well characterized in mammals, there are only few works available in the literature related to the analysis of this protein in plants. In this present work, the expression of the gene encoding TCTP was analyzed in different organs/tissues of tomato plants (Solanum lycopersicum cv. Santa Clara). A quantification performed by RT-qPCR revealed the presence of TCTP transcript in all tissues/organs analyzed, with the highest expression level found in leaves. With the exception of fruits in intermediate stage of maturation, for which a small increase on the expression was detected, there was minimal variation in the relative expression of TCTP in other organ/tissues. In parallel, the effects of the constitutive expression of TCTP were investigated using transgenic tobacco lines able to overexpress this protein at different levels (T1, T2 and T3). Seedlings of these lines, and of a non-transgenic control line, were grown in MS culture medium for 21 days. At the end of this period, the length of roots and leaves was taken and the seedlings were photographed. According to Tukey's test, the analysis of the mean root length revealed a significant difference between T1 and T3 lines when compared to the control, although the same was not observed for the T2 line. For leaves, according to Kruskal-Wallis test, there was a statistical difference between the averages of leaf growth obtained for the different lines evaluated. According to these results, we can conclude that TCTP shows an ubiquitous expression in tomato plants, with the highest expression detected in leaves, and also that its overexpression promoted a higher root and leaf development in two of three transgenic tobacco lines tested

Análise molecular de pacientes com holoprosencefalia

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise molecular de pacientes com holoprosencefalia

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Factor pronóstico del estado mutacional de BRAF en pacientes afectos de melanoma metastásico

Las mutaciones del BRAF sobreactivan la vía MAPK, estimulando así el crecimiento de las células de melanoma. En este estudio retrospectivo analizamos el estado mutacional de BRAF de 25 pacientes con melanoma metastásico con el objetivo de determinar su frecuencia y su repercusión clínica. El 52% de los pacientes presentaban mutación del BRAF, siendo estos diagnosticados a unas edades más jóvenes y en estadios más tempranos. Sin embargo, en nuestra experiencia, en los pacientes afectos de melanoma metastásico el estatus de portador de mutación de BRAF no confería una mayor supervivencia global.

Análise de polimorfismos dos genes da rota quinureniana em pacientes com meningite bacteriana.

Bacterial meningitis (BM) is still an important infectious disease causing death and disability. Invasive bacterial infections of the central nervous systems (CNS) generate some of the most powerful inflammatory responses known, which contributes to neuronal damage. The DNA microarray technology showed alterations in the kynurenine (KYN) pathway that is induced in BM and other diseases associated with inflammation, leading to brain injury. Our main aim was to search SNPs previously described in the KYN path enzymes to investigate a putative association of this SNPs with imbalanced in this pathway in patients with BM. The patients included in this study were 33 males and 24 females, with ages varying from 02 months to 68 years. SNPs were located inside of the domain conserved in KYNU, IDO, KATI and KATII. Primers were designed for analysis of SNPs already described by PIRA-PCR followed by RFLP. The analysis of KYNU+715G/A SNP found a heterozygous frequency of 0.033. We did not found the variant allele of SNP KYNU+693G/A, KATI+164T/C, KATII+650C/T and IDO+434T/G. Despite of previews studies showing the importance of KYN pathway we did not found one association of these SNPs analyzed with susceptibility or severity of MB in study population.