Serum protein profiling of lung cancer patients

Lung cancer is one of the leading causes of cancer deaths worldwide. The poor patients prognosis is largely attributable to the lack of effective early detection methods. Based on the concept that proteins and peptides can emanate from tumor to the serum, the present study aims to investigate if serum proteins pattern, assessed by a gradient polyacrylamide gel, is capable to discriminate 66 lung cancer patients from 44 healthy donors. Additionally, in a group of 10 patients and 10 healthy donors, it was also investigated by western-blot if apoptosis and metastasis-related proteins such as Bax, Bcl-2 and Hlm were present in serum. Our results showed that, in patients, protein bands of 180 kDa and 13 kDa were more frequent (Fisher, P<0.05) and a protein band of 124 kDa was more intense (Mann Whitney, P<0.05). In healthy donors a band of 158 kDa were more frequent (Fisher, P<0.05), a band of 24 kDa was more intense (Mann Whitney, P<0.05) and bands of 14 kDa and 9 kDa were together more frequent (Fisher, P<0.05) and intense (Mann Whitney, P<0.05). Bax, Bcl-2 and Hlm were not detected in serum. We conclude that changes in serum protein pattern of lung cancer patients can be detected by a simple methodology.     

Genetic engineering of allergens: future therapeutic products

Genetic engineering of allergens for specific immunotherapy should aim at the production of modified molecules with reduced IgE-binding epitopes (hypoallergens), while preserving structural motifs necessary for T cell recognition (T cell epitopes) and for induction of IgG antibodies reactive with the natural allergen (blocking antibodies). Common approaches for engineering of hypoallergens usually require knowledge of T and B cell epitopes and involve changing specific base pairs (mutated gene), introduction of a new piece of DNA into the existing DNA molecule (chimeric or hybrid gene), and deletions (truncated gene or fragments). DNA family shuffling has the advantage that it does not require a priori knowledge of structural and functional properties for efficient generation of hypoallergens. The combination of the hypoallergen concept with the Th1-inducing genetic immunization approach might be an attractive alternative for protein-based immunotherapy.     

Comparison of the effectiveness of bacterial culture, 16S rDNA directed polymerase chain reaction, and checkerboard DNA-dNA hybridization for detection of Fusobacterium nucleatum in endodontic infections

Fusobacterium nucleatum is a Gram-negative, non-spore-forming, nonmotile, obligatory anaerobic rod that is normally isolated from the oral cavity. Epidemiological studies have shown that this species is one of the most prevalent in primary root canal infections. The purpose of this study was to compare the effectiveness of bacteriological culture, 16S rDNA directed polymerase chain reaction and checkerboard DNA-DNA hybridization for detection of F. nucleatum strains in infected teeth associated with periradicular lesions. Thirteen single-root teeth from adult patients, all having carious lesions, necrotic pulps, and radiographic evidence of periradicular bone loss were included in this study. Combining all methods, the results indicated that F. nucleatum was present in approximately 31% (4 of 13) of the specimens. Incidence of F. nucleatum in root canal infections, as evaluated in this study by polymerase chain reaction, culture, and DNA-DNA hybridization, was 15.4%, 15.4%, and 10.0%, respectively. Our data demonstrated that no method used herein could be considered superior for detecting F. nucleatum directly from clinical samples. However, the small number of samples examined and the low prevalence that was observed should be considered.     

Tumour necrosis factor-alpha, interleukin-1beta and interleukin-8 induce persistent mechanical nociceptor hypersensitivity

It has been previously described that daily intraplantar (i.pl.) injections of prostaglandin E2 (PGE2) and dopamine in rats for 14 days cause the development of a persistent mechanical nociceptor hypersensitivity state lasting more than 30 days. Considering that during inflammation, the release of these hyperalgesic agents are mediated by cytokines, we investigated in the present study whether interleukin-1beta (IL-1beta), IL-8 and tumour necrosis factor-alpha (TNF-alpha) are able to induce persistent mechanical nociceptor hypersensitivity. Daily i.pl. administration of TNF-alpha, IL-1beta or IL-8 for 18 days led to persistent mechanical nociceptor hypersensitivity, which lasted at least 30 days after the cessation of treatment. The co-treatment of the animals with IL-1beta plus indomethacin, but not with atenolol, prevented the induction of persistent mechanical nociceptor hypersensitivity. The co-treatment of the animals with IL-8 plus atenolol, but not with indomethacin, prevented the induction of persistent mechanical nociceptor hypersensitivity. The daily co-treatment of TNF-alpha with either indomethacin or atenolol partially inhibited (+/-50%) the induction of persistent mechanical nociceptor hypersensitivity. However, the combined treatment with indomethacin plus atenolol abolished the induction of the persistent mechanical nociceptive hypersensitivity by TNF-alpha.A single injection of cytokines in the contralateral paws of the animals with persistent hypersensitivity caused only an acute nociceptive response. This observation, together with the demonstration of undetectable levels of immunoglobulins against TNF-alpha, IL-1beta or IL-8 in the sera of animals after the development of the persistent hypersensitivity induced by those cytokines, indicate that this event is not due to an ongoing immunological response against the cytokines. In conclusion, our results support the suggestion that IL-1beta- and IL-8-induced persistent mechanical nociceptor hypersensitivity results from the endogenous release of eicosanoids and sympathetic amines, respectively. However, TNF-alpha-induced mechanical nociceptor hypersensitivity results from the concomitant endogenous release of eicosanoids and sympathomimetic mediators.     

Studies on oogenesis and oviposition in the brown spider Loxosceles intermedia (Araneae: Sicariidae)

Loxosceles intermedia is a poisonous spider that has a wide distribution in southern Brazil, and constitutes a public health problem. In this study, the ovaries of mature females were examined by light and electron microscopy, and by histochemistry. Oocytes at all stages of development were observed in the same area of the mature ovarian wall, surrounded by a basement membrane and a proteic band. They were supported by a group of pedicular cells, which may function as follicular cells. No follicular cell was seen around the oocyte. Mature oocytes pass through the ovarian epithelial wall and are released into the ovarian lumen, covered by a vitelline membrane. The basement membrane and proteic band remain in the ovarian wall. On its way out, the oocyte is coated by proteins that will form the chorion. The presence of different coats throughout oogenesis, and at the time of egg release, is correlated with conditions that indicate fertilization occurs in the uterus lumen during oviposition.