Cloning and expression of the cDNA encoding rat granulocyte colony-stimulating factor

Granulocyte colony-stimulating factor (G-CSF) acts on precursor hematopoietic cells to control the production and maintenance of neutrophils. Recombinant G-CSF (re-G-CSF)is used clinically to treat patients with neutropenia and has greatly reduced the infection risk associated with bone marrow transplantation. Cyclic hematopoiesis, a stem cell defect characterized by severe recurrent neutropenia, occurs in man and grey collie dogs, and can be treated by administration of re-G-CSF. Availability of the rat G-CSF cDNA would benefit the use of rats as models of gene therapy for the treatment of cyclic hematopoiesis. In preliminary rat experiments, retroviral-mediated expression of canine G-CSF caused neutralizing antibody formation which precluded long-term increases in neutrophil counts. To overcome this problem we cloned the rat G-CSF cDNA from RNA isolated from skin fibroblasts. The rat G-CSF sequence shared a high degree of identity in both the coding and non-coding regions with both the murine G-CSF (85%) and human G-CSF (74%). The signal peptides of murine and human G-CSF both contained 30 amino acids (aa), whereas the deduced signal sequence for rat G-CSF possessed 21 aa. A retrovirus encoding the rat G-CSF cDNA synthesized bioactive G-CSF from transduced vascular smooth muscle cells.

Tissue distribution of a plasmid DNA encoding Hsp65 gene is dependent on the dose administered through intramuscular delivery

In order to assess a new strategy of DNA vaccine for a more complete understanding of its action in immune response, it is important to determine the in vivo biodistribution fate and antigen expression. In previous studies, our group focused on the prophylactic and therapeutic use of a plasmid DNA encoding the Mycobacterium leprae 65-kDa heat shock protein (Hsp65) and achieved an efficient immune response induction as well as protection against virulent M. tuberculosis challenge. In the present study, we examined in vivo tissue distribution of naked DNA-Hsp65 vaccine, the Hsp65 message, genome integration and methylation status of plasmid DNA. The DNA-Hsp65 was detectable in several tissue types, indicating that DNA-Hsp65 disseminates widely throughout the body. The biodistribution was dose-dependent. In contrast, RT-PCR detected the Hsp65 message for at least 15 days in muscle or liver tissue from immunized mice. We also analyzed the methylation status and integration of the injected plasmid DNA into the host cellular genome. The bacterial methylation pattern persisted for at least 6 months, indicating that the plasmid DNA-Hsp65 does not replicate in mammalian tissue, and Southern blot analysis showed that plasmid DNA was not integrated. These results have important implications for the use of DNA-Hsp65 vaccine in a clinical setting and open new perspectives for DNA vaccines and new considerations about the inoculation site and delivery system. © 2006 Coelho-Castelo et al; licensee BioMed Central Ltd.

Methods and tools for encoding the WordNet.Br sentences, concept glosses, and conceptual-semantic relations

This paper presents the overall methodology that has been used to encode both the Brazilian Portuguese WordNet (WordNet.Br) standard language-independent conceptual-semantic relations (hyponymy, co-hyponymy, meronymy, cause, and entailment) and the so-called cross-lingual conceptual-semantic relations between different wordnets. Accordingly, after contextualizing the project and outlining the current lexical database structure and statistics, it describes the WordNet.Br editing GUI that was designed to aid the linguist in carrying out the tasks of building synsets, selecting sample sentences from corpora, writing synset concept glosses, and encoding both language-independent conceptual-semantic relations and cross-lingual conceptual-semantic relations between WordNet.Br and Princeton WordNet © Springer-Verlag Berlin Heidelberg 2006.

Dietary restriction abrogates antibody production induced by a DNA vaccine encoding the mycobacterial 65 kDa heat shock protein

Background: Protein-calorie malnutrition (PCM) is the most common type of malnutrition. PCM leads to immunodeficiency and consequent increased susceptibility to infectious agents. In addition, responses to prophylactic vaccines depend on nutritional status. This study aims to evaluate the ability of undernourished mice to mount an immune response to a genetic vaccine (pVAXhsp65) against tuberculosis, containing the gene coding for the heat shock protein 65 from mycobacteria. Methods: Young adult female BALB/c mice were fed ad libitum or with 80% of the amount of food consumed by a normal diet group. We initially characterized a mice model of dietary restriction by determining body and spleen weights, hematological parameters and histopathological changes in lymphoid organs. The ability of splenic cells to produce IFN-gamma and IL-4 upon in vitro stimulation with LPS or S. aureus and the serum titer of specific IgG1 and IgG2a anti-hsp65 antibodies after intramuscular immunization with pVAXhsp65 was then tested. Results: Dietary restriction significantly decreased body and spleen weights and also the total lymphocyte count in blood. This restriction also determined a striking atrophy in lymphoid organs as spleen, thymus and lymphoid tissue associated with the small intestine. Specific antibodies were not detected in mice submitted to dietary restriction whereas the well nourished animals produced significant levels of both, IgG1 and IgG2a anti-hsp65. Conclusion: 20% restriction in food intake deeply compromised humoral immunity induced by a genetic vaccine, alerting, therefore, for the relevance of the nutritional condition in vaccination programs based on these kinds of constructs. © 2009 Ishikawa et al; licensee BioMed Central Ltd.

On the consistency conditions to braneworlds in scalar-tensor gravity for arbitrary dimensions

We derive an one-parameter family of consistency conditions to braneworlds in the Brans-Dicke gravity. The General Relativity case is recovered by taking a correct limit of the Brans-Dicke parameter. We show that it is possible to build a multiple AdS brane scenario in a six-dimensional bulk only if the brane tensions are negative. Besides, in the five-dimensional case, it is showed that no fine tuning is necessary between the bulk cosmological constant and the brane tensions, in contrast to the Randall-Sundrum model. Copyright © owned by the author(s) under the terms of the Creative Commons Attribution-NonCommercial- ShareAlike Licence.

Encoding through generalized polynomial codes

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

Light-cone fluctuations and the renormalized stress tensor of a massless scalar field

We investigate the effects of light-cone fluctuations over the renormalized vacuum expectation value of the stress-energy tensor of a real massless minimally coupled scalar field defined in a (d+1)-dimensional flat space-time with topology R×Td. For modeling the influence of light-cone fluctuations over the quantum field, we consider a random Klein-Gordon equation. We study the case of centered Gaussian processes. After taking into account all the realizations of the random processes, we present the correction caused by random fluctuations. The averaged renormalized vacuum expectation value of the stress-energy associated with the scalar field is presented. © 2013 World Scientific Publishing Company.

Massive spin-2 particle from a rank-2 tensor

Here we obtain all possible second-order theories for a rank-2 tensor which describe a massive spin-2 particle. We start with a general second-order Lagrangian with ten real parameters. The absence of lower-spin modes and the existence of two local field redefinitions leads us to only one free parameter. The solutions are split into three one-parameter classes according to the local symmetries of the massless limit. In the class which contains the usual massive Fierz-Pauli theory, the subset of spin-1 massless symmetries is maximal. In another class where the subset of spin-0 symmetries is maximal, the massless theory is invariant under Weyl transformations and the mass term does not need to fit into the form of the Fierz-Pauli mass term. In the remaining third class neither the spin-1 nor the spin-0 symmetry is maximal and we have a new family of spin-2 massive theories. © 2013 American Physical Society.

Partículas de spin-1 em D-dimensões via tensor simétrico

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The Epstein-Glaser causal approach to the light-front QED(4). II: Vacuum polarization tensor

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

Cyclic codes through B[X;(a/b)Z_0, with (a/b) in Q^{+} and b=a+1, and Encoding

Let B[X; S] be a monoid ring with any fixed finite unitary commutative ring B and is the monoid S such that b = a + 1, where a is any positive integer. In this paper we constructed cyclic codes, BCH codes, alternant codes, Goppa codes, Srivastava codes through monoid ring . For a = 1, almost all the results contained in [16] stands as a very particular case of this study.

Brazilian portuguese wordNet: a computational linguistic exercise of encoding bilingual relational lexicons

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ovarian superstimulation using FSH combined with equine chorionic gonadotropin (eCG) upregulates mRNA encoding proteins involved with LH receptor intracellular signaling in granulosa cells from Nelore cows

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