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1.
Biomed Microdevices ; 16(5): 661-70, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24831451

RESUMO

In this work a polymer lab-on-a-chip (LOC), fabricated through MEMS technology, was employed to execute a genetic protocol for the Single Nucleotide Polymorphisms (SNPs) detection. The LOC was made in Poly (methyl methacrylate) (PMMA) and has two levels: the lower one for the insertion and mixing of the reagents, the upper one for the interfacing with the DNA microarray chip. The hereditary hearing loss was chosen as case of study, since the demand for testing such a particular disorder is high and genetics behind the condition is quite clear. The Arrayed Primer EXtension (APEX) genetic protocol was implemented on the LOC to analyze the SNPs. A low density (for detection of up to 10 mutations) and a high density microarray chips (for detection of 245 mutations in 12 genes), containing the primers for the extension, were employed to carry out the APEX reaction on the LOC. Both the microarray chips provide a signal to noise ratio and efficiency comparable with a detection obtained in a conventional protocol in standard conditions. Moreover, significant reduction of the employed PCR volume (from 30 µL to 10 µL) was obtained using the low density chip.


Assuntos
Primers do DNA/química , Dispositivos Lab-On-A-Chip , Análise de Sequência com Séries de Oligonucleotídeos/instrumentação , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Polimorfismo de Nucleotídeo Único , Humanos
2.
Environ Microbiol ; 10(10): 2824-41, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18662308

RESUMO

Vetiver is the only grass cultivated worldwide for the root essential oil, which is a mixture of sesquiterpene alcohols and hydrocarbons, used extensively in perfumery and cosmetics. Light and transmission electron microscopy demonstrated the presence of bacteria in the cortical parenchymatous essential oil-producing cells and in the lysigen lacunae in close association with the essential oil. This finding and the evidence that axenic Vetiver produces in vitro only trace amounts of oil with a strikingly different composition compared with the oils from in vivo Vetiver plants stimulated the hypothesis of an involvement of these bacteria in the oil metabolism. We used culture-based and culture-independent approaches to analyse the microbial community of the Vetiver root. Results demonstrate a broad phylogenetic spectrum of bacteria, including alpha-, beta- and gamma-Proteobacteria, high-G+C-content Gram-positive bacteria, and microbes belonging to the Fibrobacteres/Acidobacteria group. We isolated root-associated bacteria and showed that most of them are able to grow by using oil sesquiterpenes as a carbon source and to metabolize them releasing into the medium a large number of compounds typically found in commercial Vetiver oils. Several bacteria were also able to induce gene expression of a Vetiver sesquiterpene synthase. These results support the intriguing hypothesis that bacteria may have a role in essential oil biosynthesis opening the possibility to use them to manoeuvre the Vetiver oil molecular structure.


Assuntos
Bactérias/classificação , Bactérias/metabolismo , Vetiveria/microbiologia , Óleos Voláteis/metabolismo , Bactérias/genética , Bactérias/isolamento & purificação , Carbono/metabolismo , Vetiveria/citologia , Vetiveria/ultraestrutura , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Perfilação da Expressão Gênica , Genes de RNAr , Microscopia , Microscopia Eletrônica , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/biossíntese , Raízes de Plantas/citologia , Raízes de Plantas/microbiologia , Raízes de Plantas/ultraestrutura , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Sesquiterpenos/metabolismo
3.
Anthropol Anz ; 72(2): 235-44, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25806538

RESUMO

Frequently, identification of individuals is problematical due to the level of associated decomposition and even more when the skeletal remains are incomplete or fragmented. The identikit, which includes a sketch or a facial reconstruction, could assist investigators with determining the identity of the decedent. Similarly, in archeology and physical anthropology it gives a realistic appearance to a historical character known only through iconography. We examined the skull of Angelo Poliziano, an Italian humanist of the 15(th) century. Previously, his facial approximation was completed in clay according to the Manchester protocol and then a duplication was prepared in ultra-realistic materials. This technique returns a long lasting 3D model of the individual and provides the perception to be in front of a real person and, although expensive, applied in forensic context could it improve the recognition of the individual.


Assuntos
Face/anatomia & histologia , Antropologia Forense/métodos , Modelos Biológicos , Adulto , Silicatos de Alumínio , Argila , Humanos , Imageamento Tridimensional , Masculino , Pessoa de Meia-Idade , Escultura , Crânio/anatomia & histologia
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