Genetic diversity and molecular phylogeny of Anaplasma marginale studied longitudinally under natural transmission conditions in Rio de Janeiro, Brazil

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

Cytotoxic and genotoxic effects of high concentrations of the immunosuppressive drugs cyclosporine and tacrolimus in MRC-5 cells

Immunosuppressive drugs are used to suppress immune system activity in transplant patients and reduce the risk of organ rejection. The present study evaluated the potential cytotoxic, genotoxic and mutagenic of the immunosuppressive drugs cyclosporine (CsA) and tacrolimus (FK-506) on normal human fibroblasts (MRC-5 cells). Based on plasma concentrations of the immunosuppressive drugs, which were obtained from the records of kidney transplant patients at the Kidney Institute of Londrina, Brazil, 11 concentrations of each immunosuppressive were chosen to evaluate cell viability using the MIT assay. From these results, CsA and FK-506 concentrations of 135, 300, 675, and 1520 ng/ml and 8, 16, 24, and 32 ng/ml, respectively, were evaluated using (i) the comet assay, (ii) the nuclear division index (NDI), (iii) the micronucleus test (CBMN) and (iv) cell proliferation curves generated by quantifying cell numbers and protein levels. In this study, 1520 to 3420 ng/ml CsA decreased cell viability after 48 h of exposure. Genotoxic effects were observed only with a concentration of 1520 ng/ml after 3 h of exposure and with concentrations of 675 and 1520 ng/ml after 24 h of exposure. Mutagenic effects were observed only for the concentration of 1520 ng/ml. FK-506 decreased cell viability after 72 h of exposure for concentrations up to 20 ng/ml; genotoxic effects were observed with concentrations up to 8 ng/ml for both treatment times (3 and 24 h) and mutagenic effects were observed with concentrations of 24 and 32 ng/ml after 24 h of treatment. The cell proliferation curves demonstrated the absence of cytostatic effects of these drugs, and these data were confirmed by the NDI analysis. Our results suggest that concentrations lower than 300 ng/ml of CsA and 16 ng/ml of FK-506 are safe for use, as they did not induce genotoxic and mutagenic damage or affect MRC-5 cell viability and proliferation. (C) 2014 Elsevier GmbH. All rights reserved.

A new heterologous fibrin sealant as scaffold to recombinant human bone morphogenetic protein-2 (rhBMP-2) and natural latex proteins for the repair of tibial bone defects

Tissue engineering has special interest in bone tissue aiming at future medical applications Studies have focused on recombinant human bone morphogenetic protein-2 (rhBMP-2) and natural latex proteins due to the osteogenic properties of rhBMP-2 and the angiogenic characteristic of fraction 1 protein (P-1) extracted from the rubber tree Hevea brasiliensis. Furthermore, heterologous fibrin sealant (FS) has been shown as a promising alternative in regenerative therapies. The aim of this study was to evaluate these substances for the repair of bone defects in rats. A bone defect measuring 3 mm in diameter was created in the proximal metaphysis of the left tibia of 60 rats and was implanted with rhBMP-2 or P-1 in combination with a new heterologous FS derived from snake venom. The animals were divided into six groups: control (unfilled bone defect), rhBMP-2 (defect filled with 5 mu g rhBMP-2), P-1 (defect filled with 5 mu g P-1), FS (defect filled with 8 mu g FS), FS/rhBMP-2 (defect filled with 8 mu g FS and 5 mu g rhBMP-2), FS/P-1 (defect filled with 8 mu g FS and 5 mu g P-1). The animals were sacrificed 2 and 6 weeks after surgery. The newly formed bone projected from the margins of the original bone and exhibited trabecular morphology and a disorganized arrangement of osteocyte lacunae. Immunohistochemical analysis showed intense expression of osteocalcin in all groups. Histometric analysis revealed a significant difference in all groups after 2 weeks (p < 0.05), except for the rhBMP-2 and FS/rhBMP-2 groups (p > 0.05). A statistically significant difference (p < 0.05) was observed in all groups after 6 weeks in relation to the volume of newly formed bone in the surgical area. In conclusion, the new heterologous fibrin sealant was found to be biocompatible and the combination with rhBMP-2 showed the highest osteogenic and osteoconductive capacity for bone healing. These findings suggest a promising application of this combination in the regeneration surgery.

Immunoexpression of ROCK-1 and MMP-9 as prognostic markers in breast cancer

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

Ultrastructure of the parotid and submandibular glands of the Old World marten (Carnivora; Mustelidae)

The morphology of the parotid and submandibular glands in the marten, a carnivore, were studied and analyzed under a transmission electron microscope. The nature of the granules in both glands, as well as in the acini and in the secretory tubules, is rather mucous. The structure of the secretory tubules is very characteristic, especially the striated ones. The myoepithelial cells are close to the acini and tubules and covered by the basement membrane separating them from the connective tissue, which enhances its epithelial origin. The cytoplasm of the basal parts of the acinar and tubular cells is abundant and separates the nucleus from the secretion granules. Although the morphology of the salivary glands of many carnivores is known, those of the parotid gland of the marten present peculiar characteristics, since they produce a rather mucous saliva and the granules, when forming, are far from the base as well as from the apex of the secretory cells. The submandibular gland contains granules of different densities, an aspect that in general resembles that of other animals.

Morphometric evaluation of the rat testis, epididymis and vas deferens following chemical sympathectomy with guanethidine

Selective chemical sympathectomy of the internal sex organs of adult male rats was undertaken by long term administration of low doses of guanethidine. The spermatogenic activity of the testis was unaffected by treatment. Examination of the vas deferens using morphometric methods revealed a marked increase in luminal area in contrast to a decrease in muscle layer area and in epithelial height. This is morphological evidence of sperm accumulation caused by a disorder in ductal contractile activity. No structural changes were observed in the epididymis. However, the concentration of spermatozoa in the sperm suspension stored in the cauda epididymidis was significantly increased in denervated rats. This result is discussed in terms of a sympathetic control of resorption mechanisms in the epididymis.

Moulting behavior in leaf-frogs of the genus phyllomedusa (Anura : hylidae)

The species belonging to the subfamily Phyllomedusinae presents physiological and behavioral adaptations to economise water such as secretion of lipids coupled with its spreading onto the skin surface through a complex behavior, adoption of diurnal torpor and secretion of uric acid. Here we describe other adaptation probably involved with water economy, registered in Phyllomedusa distincta, P. tarsius, P. tetraploidea and natural hybrids of P. distincta and P. tetraploidea - at dusk, these amphibians perform an elaborate moulting behavior that encompasses the cleaning of the body with the limbs plus gaping and body contractions. Our data show the daily moulting in P. distincta and suggest that the same mechanism occurs for the other species studied. We suppose that daily moulting improves the skin permeability to water being a characteristic of the phyllomedusines which spread lipids onto the skin surface.

Haematology of captive grey-brocket deer Mazama gouazoubira (Fischer, 1814) (Cervidae: Odocoileinae).

Reference intervals for haematologic and total plasma proteins profiles were determined for 11 physically restrained adult grey-brocket deer. Erythrocytic alterations, as discrete to moderate poikilocytosis, were observed in all animals. Red and white blood cells counts were determined by manual methodology. Obtained erythron and leukon values were slightly higher than previous reports. Statistical difference was not demonstrated between stags and hinds. No previous studies reported blood values for captive grey-brocket deer obtained by manual methods. Also, the authors suggest that future studies on deer haematology should be focused on technique evaluation and improvement, considering the unusual erythrocytic morphology.

Helminthological screening of free-ranging grey brocket deer Mazama gouazoubira Fischer, 1817 (Cervidae: Odocoileini) from Brazilian Pantanal wetlands, with considerations on Pygarginema verrucosa (Molin, 1860) Kadenatzii, 1948 (Spirocercidae: Ascaropsinae)

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

The reformulation of nonlinear complementarity problems using the Fischer-Burmeister function

A bounded-level-set result for a reformulation of the box-constrained variational inequality problem proposed recently by Facchinei, Fischer and Kanzow is proved. An application of this result to the (unbounded) nonlinear complementarity problem is suggested. © 1999 Elsevier Science Ltd. All rights reserved.

Efeito da ingestão crônica de álcool na progressão de periodontite induzida em ratos da linhagem Fischer-344

Objective: Understand the effect of chronic alcohol on the progression of periodontitis induced in Fischer-344 rats. Methods: For the study, 22 Fischer-344 rats, two months old were used, divided into groups: alcohol (n=8), ligature (n=7) and control (n=7). On the first day, the animals in the alcohol group were exposed to ingestion of a water solution containing 20% alcohol (size/size), up to day 90. After thirty days from the beginning of the experiment, the animals in the alcohol group and the ligature group were submitted to the placement of a silk thread around the right maxillary second molar. Nothing was performed on the left side, serving as control. All the groups were submitted to euthanasia 60 days after ligature placement. To assess the destruction of periodontitis, a radiographic exam was used to measure the destruction of bone height. Results: The results of the study showed that on the side in which periodontitis was induced, the group that ingested alcohol suffered an increase in destruction, with statistical differences when compared with the ligature and control groups and increased bone destruction in the ligature group when compared to control. Conclusion: Within the limitations of the study, it was concluded that chronic alcohol consumption by Fischer-344 rats led to greater progression of induced periodontitis.

Primeiro relato de atividade patogênica de Hirsutella thompsonii (Fischer) sobre o ácaro-da-erinose-da-lichia Aceria litchii (Keifer)

The litchi erineum mite, Aceria litchii (Keifer) is found in all producing regions attacking leaves and flowers of litchi plants. The mite attack young leaves and causes the erinea on leaf surface, which later become brown galls with velvety appearance. Severe attacks can cause leaf drop and destruction of branches end directly production affecting. In 2009 year it was registered a heavy infestation of the pest on litchi plants (Litchi chinensis Sonn.) in the municipality of Casa Branca, São Paulo, Brazil (2127'O; 4702'S; 679 m altitude). In June, many galls caused by mite infestation showed a mycelium of white coloration and many eriophyid dead. The fungus was identified as Hirsutella thompsonii (Fischer). The results suggest that galls may facilitate the fungus development and its permanence on the plants. Thus, the possibility of mite biological control with H. thompsonii should be investigated.

Avaliação da infecção de ratos Fischer com dua amostras geneticamente distintas de Toxoplasma gondii: cinética de anticorpos, reisolamento em camundongos e reação em cadeia pela polimerase

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