Gene therapy and its applicability in Dentistry / Terapia gênica e sua aplicabilidade na Odontologia

ABSTRACT Objective Explore scientific publications on the use of gene therapy in dentistry. Methods A bibliographic survey was carried out with articles published in the last eleven years, available on online platforms that had the highest correlation to the proposed theme. Results Since the 1960s, researchers have attempted to establish guidelines for treatments using gene therapy; however, it was only in the mid-1990s that their use in science was authorized. Gene therapy in dentistry has the ability to alter and improve genetically, through stem cells in dental tissues associated with recombinant viral vectors, therapeutic protocols against diseases that do notrespond to conventional treatments. Conclusion The perspectives of dentistry concerning this resource have been positive, mainly in the reestablishment and regeneration of tissues in pulp pathologies, periodontopathies, bone disorders, orofacial pain among others. It is known that this form of therapy still requires more research, however, in the near future, it may be a safe first option treatment in hospital and outpatient settings.

RESUMO Objetivo Explorar publicações cientificas acerca do uso da terapia gênica na Odontologia. Métodos Foi realizado um levantamento bibliográfico com artigos dos últimos dez anos disponíveis em plataformas online que continham maior correlação com o temaproposto. Resultados Desde os anos 60 pesquisadores tentam estabelecer diretrizes para tratamentos com a terapia gênica, no entanto, somente em meados dos anos 90 o seu uso na ciência foi autorizado. A terapia gênica em odontologia tem a capacidade de alterar e melhorar geneticamente, através de células-tronco em tecidos dentais e sua associação a vetores viras recombinados, protocolos terapêuticos contra enfermidades que não tem resposta aos tratamentos convencionais. Conclusão A perspectiva da odontologia diante deste recurso tem sido positiva, principalmente no reestabelecimento e regeneração de tecidos em patologias pulpares, periodopatias, distúrbios ósseos, dores orofaciais entre outros. Sabe-se que esta terapia ainda requer muitas pesquisas, entretanto em um futuro próximo poderá ser um tratamento de primeira escolha e de forma segura nos ambientes hospitalares e ambulatoriais.

Repetitive DNAs and shrink genomes: A chromosomal analysis in nine Columbidae species (Aves, Columbiformes)

Abstract An extensive karyotype variation is found among species belonging to the Columbidae family of birds (Columbiformes), both in diploid number and chromosomal morphology. Although clusters of repetitive DNA sequences play an important role in chromosomal instability, and therefore in chromosomal rearrangements, little is known about their distribution and amount in avian genomes. The aim of this study was to analyze the distribution of 11 distinct microsatellite sequences, as well as clusters of 18S rDNA, in nine different Columbidae species, correlating their distribution with the occurrence of chromosomal rearrangements. We found 2n values ranging from 76 to 86 and nine out of 11 microsatellite sequences showed distinct hybridization signals among the analyzed species. The accumulation of microsatellite repeats was found preferentially in the centromeric region of macro and microchromosomes, and in the W chromosome. Additionally, pair 2 showed the accumulation of several microsatellites in different combinations and locations in the distinct species, suggesting the occurrence of intrachromosomal rearrangements, as well as a possible fission of this pair in Geotrygon species. Therefore, although birds have a smaller amount of repetitive sequences when compared to other Tetrapoda, these seem to play an important role in the karyotype evolution of these species.

Skull shape and size variation within and between mendocinus and torquatus groups in the genus Ctenomys (Rodentia: Ctenomyidae) in chromosomal polymorphism context

Abstract We tested the association between chromosomal polymorphism and skull shape and size variation in two groups of the subterranean rodent Ctenomys. The hypothesis is based on the premise that chromosomal rearrangements in small populations, as it occurs in Ctenomys, produce reproductive isolation and allow the independent diversification of populations. The mendocinus group has species with low chromosomal diploid number variation (2n=46-48), while species from the torquatus group have a higher karyotype variation (2n=42-70). We analyzed the shape and size variation of skull and mandible by a geometric morphometric approach, with univariate and multivariate statistical analysis in 12 species from mendocinus and torquatus groups of the genus Ctenomys. We used 763 adult skulls in dorsal, ventral, and lateral views, and 515 mandibles in lateral view and 93 landmarks in four views. Although we expected more phenotypic variation in the torquatus than the mendocinus group, our results rejected the hypothesis of an association between chromosomal polymorphism and skull shape and size variation. Moreover, the torquatus group did not show more variation than mendocinus. Habitat heterogeneity associated to biomechanical constraints and other factors like geography, phylogeny, and demography, may affect skull morphological evolution in Ctenomys.

Genetic variation of the bronze locus (MC1R) in turkeys from Southern Brazil

Abstract Domestic turkeys present several color phenotypes controlled by at least five genetic loci, but only one of these has been identified precisely: the bronze locus, which turned out to be the melanocortin-1 receptor (MC1R) gene. MC1R variation is important for breeders interested in maintaining or developing different color varieties. In this study, we sequenced most of the MC1R gene from 16 White Holland (the main commercial turkey variety) and 19 pigmented turkeys from southern Brazil with two purposes. The first was to describe the MC1R diversity in White Holland turkeys, which may serve as reservoirs of genetic diversity at this locus. The second was to test whether the traditional color classification used by Brazilian breeders is related to previously known MC1R alleles. White Holland turkeys had four different haplotypes corresponding to the bronze (b+) and black-winged bronze (b1) alleles. Pigmented turkeys also had four haplotypes corresponding to the b+ and b1 alleles, but different haplotypes represent the most common b+ allele in these two groups. The black (B) allele was absent from our samples. Overall, our results suggest that white and pigmented individuals form two different populations, and that the traditional color classification used by Brazilian breeders cannot accurately predict the genotypes at the bronze locus.

Genetic structure and conservation of Mountain Lions in the South-Brazilian Atlantic Rain Forest

The Brazilian Atlantic Rain Forest, one of the most endangered ecosystems worldwide, is also among the most important hotspots as regards biodiversity. Through intensive logging, the initial area has been reduced to around 12 percent of its original size. In this study we investigated the genetic variability and structure of the mountain lion, Puma concolor. Using 18 microsatellite loci we analyzed evidence of allele dropout, null alleles and stuttering, calculated the number of allele/locus, PIC, observed and expected heterozygosity, linkage disequilibrium, Hardy-Weinberg equilibrium, F IS, effective population size and genetic structure (MICROCHECKER, CERVUS, GENEPOP, FSTAT, ARLEQUIN, ONESAMP, LDNe, PCAGEN, GENECLASS software),we also determine whether there was evidence of a bottleneck (HYBRIDLAB, BOTTLENECK software) that might influence the future viability of the population in south Brazil. 106 alleles were identified, with the number of alleles/locus ranging from 2 to 11. Mean observed heterozygosity, mean number of alleles and polymorphism information content were 0.609, 5.89, and 0.6255, respectively. This population presented evidence of a recent bottleneck and loss of genetic variation. Persistent regional poaching constitutes an increasing in the extinction risk.

Sequence variation in the melanocortin-1 receptor (MC1R) pigmentation gene and its role in the cryptic coloration of two South American sand lizards

In reptiles, dorsal body darkness often varies with substrate color or temperature environment, and is generally presumed to be an adaptation for crypsis or thermoregulation. However, the genetic basis of pigmentation is poorly known in this group. In this study we analyzed the coding region of the melanocortin-1-receptor (MC1R) gene, and therefore its role underlying the dorsal color variation in two sympatric species of sand lizards (Liolaemus) that inhabit the southeastern coast of South America: L. occipitalis and L. arambarensis. The first is light-colored and occupies aeolic pale sand dunes, while the second is brownish and lives in a darker sandy habitat. We sequenced 630 base pairs of MC1R in both species. In total, 12 nucleotide polymorphisms were observed, and four amino acid replacement sites, but none of them could be associated with a color pattern. Comparative analysis indicated that these taxa are monomorphic for amino acid sites that were previously identified as functionally important in other reptiles. Thus, our results indicate that MC1R is not involved in the pigmentation pattern observed in Liolaemus lizards. Therefore, structural differences in other genes, such as ASIP, or variation in regulatory regions of MC1R may be responsible for this variation. Alternatively, the phenotypic differences observed might be a consequence of non-genetic factors, such as thermoregulatory mechanisms.

Parâmetros hematológicos do roedor fossorial Ctenomys lami (Rodentia, Ctenomidae) no estado do Rio Grande do Sul / Hematological parameters of fossorial rodent Ctenomys lami (Rodentia, Ctenomidae) in Rio Grande do Sul, Brazil

O perfil hematológico sanguíneo foi determinado em três populações de Ctenomys lami, em áreas denominadas A e B, impactadas pela bovinocultura, e C, sem impacto antrópico, ambas no sul do Brasil. Sessenta e dois animais foram coletados ao total. Os valores de hematócrito (Ht), hemoglobina (Hb) e eritrócitos apresentaram diferenças significativas entre machos e fêmeas. Os valores médios de Ht e a Hb encontrados na espécie foram mais baixos em comparação com os de outras espécies de roedores subterrâneos, podendo estes valores estarem relacionados ao habitat de forrageio ou às características do solo. Também foram encontradas diferenças significativas nas médias de hemoglobina, CHCM e linfócitos em animais das áreas A e B em relação à área C. O valor da média dos hematócritos dos animais entre as áreas foi mais elevado nas áreas A e B, porém significativamente diferentes entre A e C. Algumas dessas alterações sugerem a relação dos valores encontrados com o estresse dos animais em relação a áreas impactadas. Variações significativas no VCM foram encontradas entre os animais das áreas A e C, e também nas plaquetas destes entre as áreas A e B. Não foram observados Corpúsculo de Kurloff no sangue dos animais analisados. Os valores hematológicos encontrados nesses espécimes de Ctenomys lami fornecem informações importantes sobre a espécie e podem ser úteis em outras pesquisas.

The hematological profile was determined in three populations of Ctenomys lami that inhabits three different areas nominated as A and B, affected by cattle production, and C, without human impact, all of them in southern Brazil, under the same geologic formation. Sixty two individuals were collected. The packed cell volume (PCV) values, hemoglobin (Hb) and red blood cell (RBC) count presents statistic significant differences between males and females. PCV and Hb values were lower in comparison with other subterranean rodents, which could be related to the food searching behavior or soil characteristic of the species. Significant statistic differences were found to for Hb, mean corpuscular hemoglobin concentration (MCHC) and lymphocytes between animals belonging to A and C areas. The PCV values were higher for animals from areas A and B, with significance only between A and C. Some of these alterations may suggest a modification on stress levels of the animals inhabiting areas A and B with anthropic alterations, or maybe just an inherited characteristic. Significant statistic differences were found either in mean corpuscular volume (MCV) between A and C areas, and platelets between A and B areas. No Kurloff cells were observed in the blood smear from analyzed individuals. The hematological values founded for Ctenomys lami are an important information for this species, which can be used for future research and management.

Evaluation of genetic variability in the collared peccary Pecari tajacu and the white-lipped peccary Tayassu pecari by microsatellite markers

In this study, the microsatellite technique was used to evaluate the genetic variability in populations of collared and white-lipped peccaries kept in captivity. Six primers developed for domestic pigs were used and amplified in both species. They revealed the presence of five polymorphic loci and one monomorphic locus. The polymorphic loci included 4 of the 16 alleles in collared peccaries, and 3 of the 10 alleles in the white-lipped peccaries. Polymorphic information content (PIC) in both species and all the loci was highly informative. The probability of paternity exclusion (PEC), if one of the parents is known, was almost as high in white-lipped peccaries (95.53 percent) as in the collared (99,48 percent). The Fst values for collared (0.042) and white-lipped (0.1387) peccaries showed that both populations are not structured. The Fis values for all loci, except ACTG2 in white-lipped peccaries (-0.0275) and in both species (0.1985 to 0.9284 in collared peccaries and 0.3621 to 0.4754 in the white-lipped), revealed a high level of homozygosis, probably caused by inbreeding. Data on heterologous amplification and genetic variability in collared and white-lipped peccaries are presented for the first time.

Genetic structure of sigmodontine rodents (Cricetidae) along an altitudinal gradient of the Atlantic Rain Forest in southern Brazil

The population genetic structure of two sympatric species of sigmodontine rodents (Oligoryzomys nigripes and Euryoryzomys russatus) was examined for mitochondrial DNA (mtDNA) sequence haplotypes of the control region. Samples were taken from three localities in the Atlantic Rain Forest in southern Brazil, along an altitudinal gradient with different types of habitat. In both species there was no genetic structure throughout their distribution, although levels of genetic variability and gene flow were high.

Polygenis (Polygenis) platensis (Jordan & Rothschild) (Siphonaptera: Rhopalopsyllidae, Rhopalopsyllinae), a new record in Brazil

Polygenis (Polygenis) platensis s. l. (Jordan & Rothschild) foi assinalada pela primeira vez no Brasil nos municípios de Capão da Canoa, Tramandaí e Osório, RS. Novos registros de hospedeiros são dados para a espécie, com a inclusão de Ctenomys flamarioni (Travi) e Ctenomys minutus (De Blainville). Variações morfológicas foram também observadas entre os espécimes coletados sobre mesma espécie de hospedeiro e em mesma localidade, evidenciando alguma hibridização entre as duas subspécies e impossibilitando a determinação do status subespecífico. O número de espécies de pulgas conhecidas para o Brasil é aumentado para 60.

Polygenis (Polygenis) platensis s. l. (Jordan & Rothschild) is recorded for the first time from Brazil in the municipalities of Capão da Canoa, Tramandaí and Osório, RS. New hostrecords for this flea include Ctenomys flamarioni (Travi) and Ctenomys minutus (De Blainville). Morphological variations were also observed among the flea specimens collected on the same species of host at the same municipality, evidencing some hybridization between the two subspecies and disabling the determination of the subspecific status. The number of known species of Brazilian Siphonaptera is now 60.

Unexpected chromosomal alterations in Tayassu tajacu (Artiodactyla: Tayassuidae) in captivity

Os animais silvestres têm sido utilizados como bioindicadores quando o ambiente é exposto a estressores químicos. Em geral, os agentes químicos podem induzir às alterações cromossômicas dos tipos falhas e quebras. Tayassu tajacu, é uma espécie aparentada dos porcos verdadeiro e apresenta uma grande estabilidade cariotípica. As únicas alterações descritas são em relação a forma do cromossomo X. Foram observadas falhas e quebras cromossômicas durante as análise citogenética. Estas alterações foram detectadas em cromossomos autossômicos. Levantamentos realizados na literatura associados as dados observados nos exemplares estudados, indicam um vermífugo, a base de ivermectina, como o possível causador dessas alterações cromossômicas.