Diferentes métodos de diagnóstico molecular na diferenciação das espécies de Sarcocystis spp

The Sarcocystis genus includes obligatory two-host life cycle protozoan parasites. It is the most numerous of the six genera of the Sarcocystidae family. The infection caused by parasites of this genus is a zoonotic and cosmopolitan disease known as sarcosistosis or sarcosporidiosis. The sarcositosis though frequently asymptomatic in its definitive hosts can be fatal in its intermediate hosts. The usual diagnoses of sarcosistosis takes place through a histological demonstration of schizonts in blood vessels and organs, and the presence of cysts in muscle tissue by necropsy or biopsy, this second method still more common and based on morphological features of the sarcocyst. However, these methods can be inadequate to a precise identification of the infector species once that, besides the genus being of numerous species, these often present similar morphological features. Another factor that makes the diagnostic more difficult is the non specificity of some Sacocystis species to their hosts. Consequently, molecular diagnostic methods have been used in order to identify the infector species and the parasite specific biological cycles, demonstrating also new species and coevolutive aspects between parasite and host. Among the most employed molecular techniques the Polimerase Chain Reaction (PCR), the nested-PCR and the Restriction Fragment Length Polymorphism (RFLP) stands out

Diagnóstico molecular da alteração mutagênica nt 230 (del4) no gene MDR1 em cães

P-glycoprotein is an adenosine triphosphate (ATP)-driven drug efflux carrier responsible for transport of xenobiotics and multiple classes of drugs, many usually use in veterinary medicine. Encoded by MDR1 gene, also referred to as ABCB1, located on chromosome 14, is expressed in many tissues with secretory or excretory functions, such as liver, kidney and intestine, where it limits drug absorption from the gut and promotes drug excretion into the bile and urine of their substrates. In 2001, a 4 base pair gene deletion mutation in the canine MDR1 gene was identified as MDR1-1▲, ABCB1-1▲, MDR1 MDR1 nt 230 (del4) and associated with an non-functional Pglycoprotein. The clinical correlation is the (hyper) sensitivity of certain dogs breeds, mostly collies, to a few classes of drugs such as anticancer drugs (doxorubicin, vincristine, vinblastine), immunosuppressants (cyclosporine), antiparasitic drugs (ivermectin, moxidectin), steroids hormones (aldosterone, cortisol, dexamethasone), antimicrobial agents (tetracycline, doxycycline, levofloxacin, ketoconazole, itraconazole), analgesics (morphine, methadone), antidiarrheals (loperamide), antiepileptic agents (phenothiazine), cardiac drugs (digoxin, diltiazem, verapamil, talinolol) and others. Dogs with homozygous MDR1 nt 230 (del4) MDR1 mutations (MDR1 - / -) have a higher predisposition to intoxication with substrates of P-gp than heterozygous (MDR1 + / -) and these are more likely than dogs homozygous nonmutant (MDR1 +/ +). After the identification of nt230 (del4) mutation, several molecular techniques have been developed for identification of mutant animals as a diagnostic method. The importance of molecular diagnosis is, after the identification of mutant animals, establish treatment protocols safe, exclude this animals from reproduction (genetic selection program) and investigating the history of adverse drugs reactions... (Complete abstract click electronic access below)

Mapeamento físico dos genes ribosomais 18S e 5S em peixes do gênero Trichomycterus (Teleostei, Trichomycteridae)

The present study aimed to cytogenetic analysis and structural and molecular level of four fish species of the genus Trichomycterus: T. diabolus, T. iheringi, T. zonatus and T. cf. mimonha collected in different river basins in Brazil. Techniques were used for classical cytogenetic (Giemsa, Silver nitrate impregnation, C-banding) and molecular with the chromosomal location of genes for 18S and 5S rDNA. All individuals examined had a diploid number 54 chromosomes and karyotype consisting of types of metacentric, submetacentric and subtelocentric. The constitutive heterochromatin identified by C-banding was observed in two small blocks in the karyotype of T. diabolus and large blocks of centromeric several pairs in chromosomal karyotypes of T. iheringi, T. zonatus and T. cf. mimonha. The Silver nitrate impregnation and hybridization with 18S rDNA probe revealed the existence of only a couple carryng nucleolar organizing regions (NORs) on the species T. diabolus, T. iheringi and T. cf mimonha, and two pairs carrying 18S rDNA in T. zonatus. The 5S rDNA was observed in interstitial position 6 of the pair in T. iheringi, the synteny with the two pair in 18S rDNA of T.diabolus in pericentromeric position of and two pairs submetacentric, one being also a case of synteny with the 18S rDNA in T. zonatus and T. cf. mimonha, this rDNA was located in the pairs 3, 18 and 25, and synteny in the 18S rDNA pair 18. Although representatives of these four species Trichomycterus present diploid number and karyotypic formula preserved, three is specific about the distribuition patterns of heterochromatin and location of rDNA sequences, indicating that chromosomal differentiation events in this group of fish are acting directly on these genomic portions

Caracterização molecular, citogenética e fenotípica de acessos do Complexo Saccharum para fins de introgressão genética

Pós-graduação em Agronomia (Genética e Melhoramento de Plantas) - FCAV

Estudo das interações de peptídeos antimicrobianos e sistemas miméticos de membrana por dinâmica molecular

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Molecular investigation of zoonotic genotypes of Giardia intestinalis isolates in humans, dogs and cats, sheep, goats and cattle in Araçatuba (Sãoo Paulo State, Brazil) by the analysis of beta-giardin gene fragments

In the period from July 2009 to October 2010, fecal samples from 61 animals and 154 humans from the municipality of Aracatuba (São Paulo State, Brazil) were studied. Fecal samples from animals were collected in the Municipal Animal Shelter and the Veterinary Hospital of the Universidade Estadual Paulista. Human fecal specimens were collected in playschools in the outskirts of the city by the private network of clinical analysis laboratories of the municipal. Diagnosis was done by optical microscopy using the Faust and Hoffmann, Pons and Janer techniques. The genotypes of Giardia intestinalis were characterized by PCR-RFLP and confirmed by sequencing the ß-giardin gene. Human specimens were positive in 25.3% (39/154) of the cases with 26.8% (36/134) of the specimens from children and 15% (3/20) from adults being positive. The frequency of G. intestinalis among the animals was 23.0% (14/61). A total of 32 isolates of G. intestinalis obtained from human feces and six from dogs and cats were characteristic of the A genotype (AI and AII/AIII). The results of this study in respect to frequency of giardiasis are similar to reported in most studies in Brazil. The prevalence observed in animal populations conforms to worldwide infection rates. G. intestinalis genotypes considered zoonotic were detected in both pets and humans from the city of Aractuba, suggesting a possible zoonotic transmission of the parasite in the northwestern region of São Paulo State. The absence of these genotypes in farm animals may imply that they are not involved in the chain of transmission to humans in this region.

Análise citogenética clássica, molecular e ultraestrutural em escorpiões da família Buthidae: um enfoque evolutivo

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

Análise celular e molecular da ação do peptídeo Ac2-26 da proteína anti-inflamatória Anexina A nas células de carcinoma de colo de útero

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização molecular, citogenética e sensibilidade a herbicidas em três populações de Rottboellia cochinchinensis em cana-de-açúcar no estado de São Paulo

Pós-graduação em Agronomia (Genética e Melhoramento de Plantas) - FCAV

Theoretical study of molecular and electronic structures of 5(1) knot systems: two-layered ONIOM calculations

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

Molecular and serological detection of Babesia Bovis and Babesia Bigemina in cattle in the Rio de Janeiro, Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Molecular detection of hemotrophic mycoplasmas among domiciled and free-roaming cats in Campo Grande, state of Mato Grosso do Sul, Brazil

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

Isolation and molecular identification of Fusobacterium nucleatum from Nigerian patients with oro-facial infections.

Fusobacterium nucleatum is one of the most common anaerobic bacteria present in the oral cavity and is often isolated from infections involving other body sites. To characterise F. nucleatum strains from patients attending a teaching hospital in Nigeria in order to provide information on the methods for accurate identification of anaerobes in clinical specimen. Fusobacterium nucleatum specie from 50 patients presenting with oro-facial infections were studied by culture on Fusobacterium selective agar and fastidious anaerobe agar. The isolates were characterised based on colonial morphology, microscopy, lipase production, susceptibility to kanamycin and colistin and resistance to vancomycin. Biochemical tests were performed using a commercial test kit. The identity of the isolates was confirmed based on molecular characterization performed using polymerase chain reaction (PCR) analysis. Forty-eight (96%) F. nucleatum isolates were obtained from the 50 patients by culture and all the isolates were identified by colonial appearance and microscopy based on their unique spindle shape with tapered ends. Only 26 (54.2%) of the 48 isolates were identified by commercial API 20A test kit while PCR confirmed the identity of all the isolates. Anaerobes are involved in human infections and their study is quite cumbersome due to tedious nature and high cost of the techniques involved. Cultural method is reliable in the isolation and identification of F. nucleatum species. PCR is a rapid and simple method that can complement the phenotypic identification of anaerobes and would assist in their full identification.

Anti-inflammatory drug design by molecular hybridization approach

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

Nanoestruturas de GaN crescidas pelas técnicas de epitaxia por magnetron sputtering e epitaxia por feixe molecular

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