Gene expression profiling reveals molecular marker candidates of laryngeal squamous cell carcinoma

Laryngeal squamous cell carcinoma is very common in head and neck cancer, with high mortality rates and poor prognosis. In this study, we compared expression profiles of clinical samples from 13 larynx tumors and 10 non-neoplastic larynx tissues using a custom-built cDNA microarray containing 331 probes for 284 genes previously identified by informatics analysis of EST databases as markers of head and neck tumors. Thirty-five genes showed statistically significant differences (SNR >= 11.01, p <= 0.001) in the expression between tumor and non-tumor larynx tissue samples. Functional annotation indicated that these genes are involved in cellular processes relevant to the cancer phenotype, such as apoptosis, cell cycle, DNA repair, proteolysis, protease inhibition, signal transduction and transcriptional regulation. Six of the identified transcripts map to intronic regions of protein-coding genes and may comprise non-annotated exons or as yet uncharacterized long ncRNAs with a regulatory role in the gene expression program of larynx tissue. The differential expression of 10 of these genes (ADCY6, AES, AL2SCR3, CRR9, CSTB, DUSP1, MAP3K5, PLAT, UBL1 and ZNF706) was independently confirmed by quantitative real-time RT-PCR. Among these, the CSTB gene product has cysteine protease inhibitor activity that has been associated with an antimetastatic function. Interestingly, CSTB showed a low expression in the tumor samples analyzed (p<0.0001). The set of genes identified here contribute to a better understanding of the molecular basis of larynx cancer, and provide candidate markers for improving diagnosis, prognosis and treatment of this carcinoma.

Electroanalytical Method for the Analysis of Methyldopa In Pharmaceutical Tablets Using a Crude Extract of Laccase from Pycnoporus sanguineus as Oxidizing Agent

Several colorimetric and chromatographic methods have been used for the identification and quantification of methyldopa (MA) in pharmaceutical formulations and clinical samples. However, these methods are time- and reagent-consuming, which stimulated our efforts to develop a simple, fast, and low-cost alternative method. We carried out an electroanalytical method for the determination of MA in pharmaceutical formulations using the crude enzymatic extract of laccase from Pycnoporus sanguineus as oxidizing agent. This method is based on the biochemical oxidation of MA by laccase (LAC), both in solution, followed by electrochemical reduction on glassy carbon electrode surface. This method was employed for the determination of MA in pure and pharmaceutical formulations and compared with the results obtained using the official method. A wide linear curve from 23 x 10(-5) to 1 x 10(-4) mol L(-1) was found with a detection limit calculated from 43 x 10(-6) mol L(-1).

Fluoxetine and norfluoxetine analysis by direct injection of human plasma in a column switching liquid chromatographic system

A column switching LC method is presented for the analysis of fluoxetine (FLU) and norfluoxetine (NFLU) by direct injection of human plasma using a lab-made restricted access media (RAM) column. A RAM-BSA-octadecyl silica (C-18) column (40 min x 4.6 mm, 10 mu m) is evaluated in both backflush and foreflush elution modes and coupled with a C-18 lab-made (50 mm x 4.6 mm, 3 pm) analytical column in order to perform online sample preparation. Direct injection of 100 mu L, of plasma samples is possible with the developed approach. In addition, reduction of sample handling is obtained when compared with traditional liquid-liquid extraction (LLE) and SPE. The total analysis time is around 20 min. A LOQ of 15 ng/mL is achieved in a concentration range of 15-500 ng/mL, allowing the therapeutic drug monitoring of clinical samples. The precision values achieved are lower than 15% for all the evaluated points with adequate recovery and accuracy. Furthermore, no matrix interferences are found in the analysis and the proposed method shows to be an adequate alternative for analysis of FLU in plasma.

Avaliação da atividade antimicrobiana de sistemas emulsionados contendo óleos naturais para o tratamento de infecções cutâneas

Natural oils have shown a scientific importance due to its pharmacological activity and renewable character. The copaiba (Copaifera langsdorffii) and Bullfrog (Rana catesbeiana Shaw) oils are used in folk medicine particularly because the anti-inflammatory and antimicrobial activities. Emulsion could be eligible systems to improve the palatability and fragrance, enhance the pharmacological activities and reduce the toxicological effects of these oils. The aim of this work was to investigate the antimicrobial activity of emulsions based on copaiba (resin-oil and essential-oil) and bullfrog oils against fungi and bacteria which cause skin diseases. Firstly, the essential oil was extracted from copaiba oil-resin and the oils were characterized by gas chromatography coupled to a mass spectrometry (GC-MS). Secondly, emulsion systems were produced. A microbiological screening test with all products was performed followed (the minimum inhibitory concentration, the bioautography method and the antibiofilm determination). Staphylococcus aureus, S. epidermidis, Pseudomonas aeruginosa, Candida albicans, C. parapsilosis, C. glabrata, C. krusei and C. tropicalis American Type Culture Collection (ATCC) and clinical samples were used. The emulsions based on copaiba oil-resin and essential oil improved the antimicrobial activity of the pure oils, especially against Staphylococcus e Candida resistant to azoles. The bullfrog oil emulsion and the pure bullfrog oil showed a lower effect on the microorganisms when compared to the copaiba samples. All the emulsions showed a significant antibiofilm activity by inhibiting the cell adhesion. Thus, it may be concluded that emulsions based on copaiba and bullfrog oils are promising candidates to treatment of fungal and bacterial skin infections

Prevalence of newcastle disease virus in broiler chickens (Gallus gallus) in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Direct Detection of Mycobacterium bovis in Bovine Lymph Nodes by PCR

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A novel in situ Polymerase chain reaction hybridisation assay for the direct detection of bovine herpesvirus type 5 in formalin-fixed, paraffin-embedded tissues

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

Occurrence of yeasts, enterococci and other enteric bacteria in subgingival biofilm of HIV-positive patients with chronic gingivitis and necrotizing periodontitis

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

Atividade antifúngica de óleos essenciais frente a amostras clínicas de Candida albicans isoladas de pacientes HIV positivos

Objetivou-se avaliar a atividade antifúngica dos óleos essenciais de Ocimum basilicum L. (manjericão), Cymbopogon martinii L. (palmarosa), Thymus vulgaris L. (tomilho) e Cinnamomum cassia Blume (canela da china) sobre cepas de Candida albicans isoladas de pacientes HIV positivos e cepa padrão (ATCC 76845). Quinze amostras clínicas de C. albicans (C1-C15) foram repicadas em ágar Sabouraud Dextrose, para confecção de suspensões em solução salina estéril (0,9%) contendo 1,5 x 10(6) UFC mL-1. As emulsões dos óleos essenciais foram preparadas em água destilada estéril e tween 80, com concentrações variando entre 1024 µg mL-1 e 4 µg mL-1. A ação antifúngica foi determinada por meio da Concentração Inibitória Mínima (CIM) utilizando-se a técnica da microdiluição. Foram utilizados como controles positivos a nistatina e o miconazol (50 µg mL-1). Os testes foram realizados em triplicata, sendo a CIM, a menor concentração capaz de inibir o crescimento das leveduras, observada por método visual de acordo com a turvação do meio de cultura. Para C. albicans (ATCC 76845), a CIM do óleo essencial de C. cassia foi 64 µg mL-1, enquanto para óleo de C. martinii foi 1024 µg mL-1. Para as cepas clínicas, verificou-se que a CIM de C. cassia para 80% das cepas foi 64 µg mL-1, sendo a variação dos valores da CIM entre 128 µg mL-1 e 64 µg mL-1. Observou-se que para 66,6% das amostras clínicas, a CIM de C. martinii foi 612 µg mL-1. Constatou-se que a nistatina não apresentou atividade frente às cepas clínicas (C1-C15), enquanto a atividade antifúngica do miconazol foi verificada em 100% das amostras. Não se constatou atividade antimicrobiana dos óleos essenciais de O. basilicum e T. vulgaris, nas concentrações avaliadas. Concluiu-se que os óleos essenciais de C. cassia e C. martinii, em diferentes concentrações, apresentam atividade antifúngica sobre cepas de C. albicans isoladas de pacientes HIV positivos e cepa padrão (ATCC 76845). Entretanto não foi observada inibição antimicrobiana para os óleos de O. basilicum e T. vulgaris.

Gene expression profiling reveals molecular marker candidates of laryngeal squamous cell carcinoma

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

Identification of Mycobacteria by Thin Layer Chromatographic Analysis of Mycolic Acids and Conventional Biochemical Method: Four Years of Experience

Mycolic acids analysis by thin-layer chromatography (TLC) has been employed by several laboratories worldwide as a method for fast identification of mycobacteria. This method was introduced in Brazil by our laboratory in 1992 as a routine identification technique. Up to the present, 861 strains isolated were identified by mycolic acids TLC and by standard biochemical tests; 61% out of these strains came as clinical samples, 4% isolated from frogs and 35% as environmental samples. Mycobacterium tuberculosis strains identified by classical methods were confirmed by their mycolic acids contents (I, III and IV). The method allowed earlier differentiation of M. avium complex - MAC (mycolic acids I, IV and VI) from M. simiae (acids I, II and IV), both with similar biochemical properties. The method also permitted to distinguish M. fortuitum (acids I and V) from M. chelonae (acids I and II) , and to detect mixed mycobacterial infections cases as M. tuberculosis with MAC and M. fortuitum with MAC. Concluding, four years experience shows that mycolic acids TLC is an easy, reliable, fast and inexpensive method, an important tool to put together conventional mycobacteria identification methods.

Epidemiologia do transtorno obsessivo-compulsivo: uma revisão

Os inquéritos populacionais são importantes, pois amostras clínicas tendem a apresentar vieses de seleção. Aspectos sociodemográficos e relacionados à própria condição mórbida podem interferir na procura por tratamento. Pela natureza egodistônica do transtorno obsessivo-compulsivo, seus portadores tendem a ocultar o problema, podendo não procurar ou demorar a procurar tratamento. Porém, a maior parte do conhecimento atual sobre o transtorno obsessivo-compulsivo advém de amostras clínicas, que não representam a totalidade dos casos. Foi feita uma revisão convencional da literatura através do Medline, PsicoInfo e Lilacs de inquéritos populacionais sobre o transtorno obsessivo-compulsivo, cobrindo o período de 1980 a 2004, utilizando-se como palavras-chave epidemiologia, transtorno obsessivo-compulsivo, inquéritos populacionais e prevalência. Estudos realizados em diferentes países indicam para o transtorno obsessivo-compulsivo uma prevalência atual em torno de 1,0% e ao longo da vida de 2,0 a 2,5%. Diferentemente de amostras clínicas, em quase todas as amostras populacionais há predomínio de mulheres e portadores que têm apenas obsessões. A freqüente comorbidade com outros transtornos mentais, particularmente depressão e outros transtornos ansiosos, repete-se em casos da população geral, que apresentam ainda uma associação com abuso de substâncias. Muitos portadores não estão em tratamento, particularmente os casos puros. Indicadores de incapacitação funcional demonstram um considerável impacto negativo do transtorno obsessivo-compulsivo. É preciso melhorar o conhecimento da população e dos profissionais de saúde sobre os sintomas do transtorno obsessivo-compulsivo para aumentar a procura de atendimento, assim como a correta identificação e abordagem terapêutica deste grave problema de saúde.

Phenotypic methods and commercial systems for the discrimination between C-albicans and C-dubliniensis

Candida dubliniensis is a recently described Candida species associated with oral candidosis that exhibits a high degree of phenotypic similarity to Candida albicans. However, these species show differences in levels of resistance to antimycotic agents and ability to cause infections. Therefore, accurate clinical identification of C. dubliniensis and C. albicans species is important in order to treat oral candidal infections. Phenotypic identification methods are easy-to-use procedures for routine discrimination of oral isolates in the clinical microbiology laboratory. However, C. dubliniensis may be so far underreported in clinical samples because most currently used identification methods fail to recognize this yeast. Phenotypic methods depend on growth temperature, carbon source assimilation, chlamydospore and hyphal growth production, positive or negative growth on special media and intracellular enzyme production, among others. In this review, some phenotypic methods are presented with a special emphasis on the discrimination of C. dubliniensis and C. albicans.

Padrão de sensibilidade de cepas de Enterobacter spp isoladas na região de Presidente Prudente - SP

Enterobacter spp. are considered important causing agents of infection, specially in hospitalized individuals. The natural resistance of these microorganisms and the great facility to develop resistance to new antimicrobial agents make this genus an important object of study. In this work, 176 strains isolated from various clinical samples were used from hospitalized patients (University Hospital Domingos Leonardo Cerávolo) and from clinic patients (Clinical Laboratory from Unoeste), both situated in Presidente Prudente - SP. E. cloacae (78.9%), E. aerogenes (7.9%) and E. (pantoea) agglomerans (3.9%) were the ones more frequently isolated. Eleven antimicrobial agents were tested by the disk diffusion method and around 90% of the strains presented resistance to the cephalotin, ampicillin and cefaclor. Strains of E. (pantoea) agglomerans presented wide profile of sensibility However one strain of E. cloacae presented resistance to all the antimicrobial agents. The antimicrobial agents with greater inhibitory activity were imipenem and cefepime, for this reason these antimicrobial agents could be the treatment of choice in emergencial therapeutic. This emergencial therapy can be applied with relative security, whereas the data obtained in this study show homogeneity in the profile of sensibility to these antimicrobial agents, independent of the infection site and from the isolated species. The ESBL enzyme could not be detected in no one of the strains by the double diffusion test.

Frequency and genetic diversity of the MAT1 locus of Histoplasma capsulatum isolates in Mexico and Brazil

The MAT1-1 and MAT1-2 idiomorphs associated with the MAT1 locus of Histoplasma capsulatum were identified by PCR. A total of 28 fungal isolates, 6 isolates from human clinical samples and 22 isolates from environmental (infected bat and contaminated soil) samples, were studied. Among the 14 isolates from Mexico, 71.4% (95% confidence interval [95% CI], 48.3% to 94.5%) were of the MAT1-2 genotype, whereas 100% of the isolates from Brazil were of the MAT1-1 genotype. Each MAT1 idiomorphic region was sequenced and aligned, using the sequences of the G-217B (+mating type) and G-186AR (-mating type) strains as references. BLASTn analyses of the MAT1-1 and MAT1-2 sequences studied correlated with their respective+ and-mating type genotypes. Trees were generated by the maximum likelihood (ML) method to search for similarity among isolates of each MAT1 idiomorph. All MAT1-1 isolates originated from Brazilian bats formed a well-defined group; three isolates from Mexico, the G-217B strain, and a subgroup encompassing all soil-derived isolates and two clinical isolates from Brazil formed a second group; last, one isolate (EH-696P) from a migratory bat captured in Mexico formed a third group of the MAT1-1 genotype. The MAT1-2 idiomorph formed two groups, one of which included two H. capsulatum isolates from infected bats that were closely related to the G-186AR strain. The other group was formed by two human isolates and six isolates from infected bats. Concatenated ML trees, with internal transcribed spacer 1 (ITS1) -5.8S-ITS2 and MAT1-1 or MAT1-2 sequences, support the relatedness of MAT1-1 or MAT1-2 isolates. H. capsulatum mating types were associated with the geographical origin of the isolates, and all isolates from Brazil correlated with their environmental sources. © 2013, American Society for Microbiology. All Rights Reserved.