Cysteine proteinase inhibitors regulate human and mouse osteoclastogenesis by interfering with RANK signaling

The cysteine proteinase inhibitor cystatin C inhibited RANKL-stimulated osteoclast formation in mouse bone marrow macrophage cultures, an effect associated with decreased mRNA expression of Acp5, Calcr, Ctsk, Mmp9, Itgb3, and Atp6i, without effect on proliferation or apoptosis. The effects were concentration dependent with half-maximal inhibition at 0.3 μM. Cystatin C also inhibited osteoclast formation when RANKL-stimulated osteoclasts were cultured on bone, leading to decreased formation of resorption pits. RANKL-stimulated cells retained characteristics of phagocytotic macrophages when cotreated with cystatin C. Three other cysteine proteinase inhibitors, cystatin D, Z-RLVG-CHN2 (IC50 0.1 μM), and E-64 (IC 50 3 μM), also inhibited osteoclast formation in RANKL-stimulated macrophages. In addition, cystatin C, Z-RLVG-CHN2, and E-64 inhibited osteoclastic differentiation of RANKL-stimulated CD14+ human monocytes. The effect by cystatin C on differentiation of bone marrow macrophages was exerted at an early stage after RANKL stimulation and was associated with early (4 h) inhibition of c-Fos expression and decreased protein and nuclear translocation of c-Fos. Subsequently, p52, p65, IκBα, and Nfatc1 mRNA were decreased. Cystatin C was internalized in osteoclast progenitors, a process requiring RANKL stimulation. These data show that cystatin C inhibits osteoclast differentiation and formation by interfering intracellularly with signaling pathways downstream RANK. © FASEB.

Epidemiology of human immunodeficiency virus-visceral leishmaniasis-co-infection

In Brazil, the rates of mother-to-child-transmission (MTCT) of human immunodeficiency virus (HIV) decreased from 20% to 1-2% in some regions. However, the country contains 90% of individuals infected with visceral leishmaniasis (VL) in Latin America, and the west region of São Paulo state faces an alarming expansion of the disease. We describe the epidemiological aspects of the expanding infection of VL and a case report of an HIV-VL-co-infected child from the west region of São Paulo state. The patient was an AIDS-C3 with low levels of CD4, high viral load, severe diarrhea, oral and perineal candidiasis, severe thrombocytopenia, and protein-caloric malnourishment. She evolved with sepsis, renal and cardiac failure. An rK rapid diagnosis test, indirect fluorescent antibody test (IFAT), and bone marrow aspirate were performed for VL. Her symptoms improved significantly after liposomal amphotericin B administration. From the 45 municipalities that compose the Regional Health Department of Presidente Prudente, Lutzomyia longipalpis vectors were found in 58% of them. VL infected dogs were found in 33% of those municipalities, infected dogs and humans were found in 29%, 20% are starting and 33% of the municipalities are preparing VL investigation. It is likely, in this patient, that VL advanced the clinical progression of the HIV disease and the development of AIDS severity. Supported by favorable conditions, the region becomes a new frontier of VL in Brazil. © 2013.

Lentinula edodes (Shiitake) modulates chemically induced mutagenesis by enhancing pitting

This study was undertaken to understand how Lentinula edodes modulates in vivo mutagenesis induced by alkylating agents in bone marrow and peripheral blood as described in our previous article. Male Swiss mice were pretreated for 15 consecutive days with aqueous extracts prepared from L. edodes, after which, the number of circulating blood cells, normal erythroid bone marrow cell cycling, and phagocytosis of micronucleated reticulocyte (MNRET) and activation of spleen macrophages were assessed. The results indicate that the antimutagenicity seen in bone marrow and peripheral blood is exerted by distinct compounds with different actions. The antimutagenic effect in bone marrow is exerted by compounds subject to degradation at deep-freeze storage temperature of -20 C. On the other hand, compounds responsible for antimutagenicity in peripheral blood are not subject to degradation at -20 C. The results also indicate that the antimutagenic action in peripheral blood leading to the reduction of circulating MNRET occurs in the spleen primarily through a phagocytic activity due to higher macrophage numbers and probably not due to the enhanced activation state of individual cells. © Mary Ann Liebert, Inc.

Interaction between mesenchymal stem cells and Ti-30Ta alloy after surface treatment

In this study, in vitro cytocompatibility was investigated in the Ti-30Ta alloy after two kinds of surfaces treatments: alkaline and biomimetic treatment. Each condition was evaluated by scanning electron microscopy/energy-dispersive X-ray spectroscopy. Cellular adhesion, viability, protein expression, morphology, and differentiation were evaluated with Bone marrow stromal cells (MSCs) to investigate the short and long-term cellular response by fluorescence microscope imaging and colorimetric assays techniques. Two treatments exhibited similar results with respect to total protein content and enzyme activity as compared with alloy without treatment. However, it was observed improved of the biomineralization, bone matrix formation, enzyme activity, and MSCs functionality after biomimetic treatment. These results indicate that the biomimetic surface treatment has a high potential for enhanced osseointegration. © 2013 Wiley Periodicals, Inc.

Histologic Characterization of Engineered Tissues in the Canal Space of Closed-apex Teeth with Apical Periodontitis

Introduction: The aim of this study was to investigate the capacity of endodontic regenerative procedures combining an induced blood clot, platelet-rich plasma (PRP), and bone marrow aspirate (BMA) to regenerate dental pulp in canine closed-apex necrotic teeth. Methods: Apical periodontitis was induced in 20 upper and lower premolars of 2 dogs. After biomechanical preparation, enlargement to a #60 file, and disinfection with a triantibiotic paste for 28 days, the roots were randomly assigned to 4 treatment groups: blood clot (BC), BC + PRP gel, BC + BMA gel, and BC + BMA/PRP gel. Negative controls were also included. After a 3-month follow-up period, the animals were killed. Results: Histologic analysis showed the presence of newly formed vital tissues (connective, cement-like, and bone-like tissue) in 23 of the 32 treated roots (71.87%). There was no statistically significant difference between the treatment groups. Conclusions: New vital tissues were formed and characterized as connective, cementum-like, or bone-like, but not as pulp-like tissue; PRP and/or BMA did not improve the tissue ingrowth. © 2013 American Association of Endodontists.

Avaliação do perfil de expressão gênica de células CD34+ e células CD CD133+ isoladas de medula óssea e de sangue de cordão umbilical

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação dos efeitos citotóxicos, genotóxicos, mutagênicos, antigenotóxicos, antimutagênicos e anticarcinogênicos do Lupinus albus em camundongos Mus musculus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização do modelo B6D2F1 para obesidade, diabetes e síndrome metabólica e sua potencialidade para terapia celular em lesão aguda

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

Investigação do potencial genotóxico do extrato de Garcinia achachairu in vivo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Investigação do potencial genótóxico e antigenotóxico do extrato de Brassica oleracea in vivo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Identificação e análise de polimorfismos no gene que codifica a Lectina ligante de Manose (MBL) na espécie Canis lúpus familiaris

Pós-graduação em Biologia Geral e Aplicada - IBB

Clastogenicidade e /ou aneugenicidade do hormônio androgênico nandrolona (Deca-Durabolin®) em camundongos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Proposta de metodologia para o gerenciamento de fonte radioativa e estimativa de dose efeiva em pacientes submetidos à terapia com iodeto de sódio I-131

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Validação de anticorpo monoclonal anti-CD45 obtido in house para utilização em citometria de fluxo e imuno-histoquímica

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Mielopatia infiltrativa por tumores não-hematológicos

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB