Pigmentação testicular em Physalaemus nattereri (Steindachner) (Amphibia, Anura) com observações anatômicas sobre o sistema pigmentar extracutâneo

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

Comparative study between fibrin glue and platelet rich plasma in dogs skin grafts

OBJETIVO: Comparar a cola de fibrina (Tissucol®) e o plasma rico em plaquetas em enxertos cutâneos de espessura completa em malha em cães. MÉTODOS: Foram utilizados 18 cães, distribuídos em dois grupos, cola de fibrina (CF) e plasma rico em plaquetas (PRP). em todos os animais foi realizado um enxerto cutâneo de 3x3 cm, em malha de espessura completa. No membro esquerdo foi colocado o biomaterial entre o enxerto e o leito receptor, cada qual em seu grupo, o membro direito serviu como grupo controle. Todos os animais foram avaliados clinicamente a cada 48 horas até o décimo quarto dia, através das variáveis: exsudação, coloração, edema e aspecto cosmético; histologicamente em três animais, no terceiro, sétimo e décimo quarto dia de pós-operatório através das variáveis: fibroblastos, colágeno, tecido de granulação, integração-aderência microscópica e inflamação aguda. RESULTADOS: Avaliações clínicas demonstraram que o grupo CF apresentou melhor escores em todas variáveis quando comparado com o grupo PRP. Nas avaliações histológicas o grupo PRP apresentou maior presença de fibroblastos ao sétimo e décimo quarto dia. CONCLUSÃO: A cola de fibrina foi clinicamente superior ao grupo plasma rico em plaquetas quando usados em enxertos cutâneos de espessura completa em cães.

Análise histológica e morfométrica da área cruenta tratada com membrana de silicone: estudo em ratos

Background. Thin study evaluated the effects of a silicone membrane on the treatment of the raw flesh area in wounds.METHODS. The experimental study was carried out with 30 male Wistar rats divided into three groups: in the G(G) group, the raw area was treated with gauze, in the G(H) group with homograft, and in the G(S) group with a silicone membrane. Animals were sacrificed at fourteen days of postoperative. The studied attributes were body mass variation, histological study quantifed by morphometric analysis evaluating the number of neovessels, fibroblasts, collagen fibers, leucocytes, monocytes; and using a micrometric ruler measurement of the raw area thickness. Data were then submitted to statistical analysis.RESULTS. There was no significant difference between the animals' mass (p=0.0685). Predominance of neovessels (p<0.01), fibroblasts (p<0.001) and thickness of the raw flesh were observed in G(S) animals. There was predominance of leucocytes (p<0.021) and monocytes (p<0.0001) in G(H) animals. Also, no significant difference between the groups as for collagen fibers (p=0.0536) was observed.CONCLUSION. The silicone membrane promoted granulation tissue with a large number of neovessels, fibroblasts and greater thickness.

Identification and characterization of novel mutations of the aspartoacylase gene in non-Jewish patients with Canavan disease

Canavan disease, an inherited leukodystrophy, is caused by mutations in the aspartoacylase (ASPA) gene. It is most common among children of Ashkenazi Jewish descent but has been diagnosed in many diverse ethnic groups. Two mutations comprise the majority of mutant alleles in Jewish patients, while mutations in the ASPA gene among non-Jewish patients are different and more diverse. In the present study, the ASPA gene was analysed in 22 unrelated non-Jewish patients with Canavan disease, and 24 different mutations were found. of these,14 are novel, including five missense mutations (E24G, D68A, D249V, C152W, H244R), two nonsense mutations (Q184X, E214X), three deletions (923delT, 33del13, 244delA), one insertion mutation (698insC), two sequence variations in one allele ([10T>G; 11insG]), an elimination of the stop codon (941A>G, TAG-->TGG, X314W), and one splice acceptor site mutation (IVS1 - 2A>T). The E24G mutation resulted in substitution of an invariable amino acid residue (Glu) in the first esterase catalytic domain consensus sequence. The IVS1 - 2A>T mutation caused the retention of 40 nucleotides of intron 1 upstream of exon 2. The results of transient expression of the mutant ASPA cDNA containing these mutations in COS-7 cells and assays for ASPA activity of patient fibroblasts indicated that these mutations were responsible for the enzyme deficiency. In addition, patients with the novel D249V mutation manifested clinically at birth and died early. Also, patients with certain other novel mutations, including C152W, E214X, X314W, and frameshift mutations in both alleles, developed clinical manifestations at an earlier age than in classical Canavan disease.

Melanocytes in the testes of Eupemphix nattereri (Anura, Leiuperidae): Histological, stereological, and ultrastructural aspects

Ectothermic vertebrates have a well-developed system of melanin-containing cells, which localize in several organs and tissues and compose an extracutaneous pigmentary system. This research aimed at characterizing histological and ultrastructural patterns of pigmented cells in the testes of the anura Eupemphix nattereri (Steindachner, 1963), including the stereological and quantitative evaluation of this cell type in the gonads. Ten adult males were collected in Nova Itapirema, São Paulo, Brazil, and submitted to morphological studies with light and transmission electron microscopy. The testis presents a great number of large cells with many brown granules and long cytoplasmic processes. The pigmented cells found in the testis are structurally similar to melanocytes, characterized by large amounts of melanosomes. The cells may be in intimate contact with the same cell type, with myoid cells surrounded by a large amount of collagen fibers, Leydig cells, and next to fibroblasts. The distribution and amount of extracutaneous melanocytes is variable when other organs and membranes are analyzed, allowing the establishment of species-specific patterns for the extracutaneous pigmentary system.

Alloxan-Induced Diabetes Triggers the Development of Periodontal Disease in Rats

Background. Periodontal disease in diabetic patients presents higher severity and prevalence; and increased severity of ligature-induced periodontal disease has been verified in diabetic rats. However, in absence of aggressive stimuli such as ligatures, the influence of diabetes on rat periodontal tissues is incompletely explored. The aim of this study was to evaluate the establishment and progression of periodontal diseases in rats only with diabetes induction. Methodology/Principal Findings. Diabetes was induced in Wistar rats (n = 25) by intravenous administration of alloxan (42 mg/kg) and were analyzed at 1, 3, 6, 9 and 12 months after diabetes induction. The hemimandibles were removed and submitted to radiographical and histopathological procedures. A significant reduction was observed in height of bone crest in diabetic animals at 3, 6, 9 and 12 months, which was associated with increased numbers of osteoclasts and inflammatory cells. The histopathological analyses of diabetic rats also showed a reduction in density of collagen fibers, fibroblasts and blood vessels. Severe caries were also detected in the diabetic group. Conclusions/Significance. The results demonstrate that diabetes induction triggers, or even co-induces the onset of alterations which are typical of periodontal diseases even in the absence of aggressive factors such as ligatures. Therefore, diabetes induction renders a previously resistant host into a susceptible phenotype, and hence diabetes can be considered a very important risk factor to the development of periodontal disease.

THE ESSENTIAL ROLE OF ZINC IN GROWTH

Zinc is known to play a relevant role in growth and development. The basic mechanisms of action of this trace element are intimately linked to the structure and action of countless enzymes involved in many different metabolic processes. In this respect, when zinc specifically acts on cartilage growth it is involved in multiple enzymatic reactions which make this a multifactorial event. Thus, we may divide the actions of zinc into three distinct types: 1) action on taste and smell acuity, appetite regulation, and food consumption and regulation; 2) action on DNA and RNA synthesis stimulating a) cell replication and differentiation of chondrocytes, osteoblasts and fibroblasts; b) cell transcription culminating in the synthesis of somatomedin-C (liver), alkaline phosphatase, collagen and osteocalcin (bone), and c) protein, carbohydrate and lipid metabolism, that is intimately related to the mechanisms of smell, taste, appetite, and food consumption and utilization; 3) action on hormonal mediation by participating in a) GH synthesis and secretion in somatomammotroph cells, b) the action of GH on liver somatomedin-C production, and c) somatomedin-C activation in bone cartilage. In addition to these multiple functions, zinc also interacts with other hormones somehow related to bone growth such as testosterone, thyroid hormones, insulin, and vitamin D-3.On the basis of the above considerations, we conclude that the integration of these mechanisms contributes to the perfect physiological functioning of bone. Tn the presence of zinc deficiency, this homeostasis is impaired, causing the weight-height deficiency detected in several species studied, the human species in particular.

Experimental study of progressive facial hemiatrophy: effects of cervical sympathectomy in animals

Progressive facial hemiatrophy (Romberg's syndrome) is of unknown cause and uncertain pathogenesis. The main pathogenetic hypotheses are: sympathetic system alterations, localized scleroderma, trigeminal changes, possibly of genetic origin. To test the hypothesis of sympathetic system alterations, we designed an experimental model with ablation of the superior cervical sympathetic ganglion in rabbits, cats and dogs. All the animals were operated upon when 30 days old and were examined monthly for 1 year. During this period localized alopecia, corneal ulceration, keratitis, strabismus, enophthalmos, ocular atrophy, hemifacial atrophy and slight bone atrophy on the side of the sympathectomy were observed. Thus, cervical sympathectomy reproduces in animals the principal clinical alterations of Romberg's syndrome. Our data suggest that the sympathetic system is involved in the pathogenesis of this syndrome.

EFFECT OF THE HELIUM-NEON LASER ON THE HEALING OF EXTRACTION WOUNDS - A HISTOLOGICAL STUDY IN RATS

A histological study on the healing of extraction wounds following laser irradiation using a He-Ne laser, was carried out in rats. The results suggest that this mode of treatment has a beneficial effect on initial bony wound healing. Proliferation of fibroblasts and formation of trabecular osteoid tissue were found to be more prominent within the irradiated group. The deposition of led in the newly formed bone suggests swifter ossification within the irradiated group.

Thermal Characterization and Cytotoxicity of Complexes Formed by Papain and Cyclodextrin

Papain is a proteolytic enzyme with restricted applications due to its limited stability. Cyclodextrins are widely used in pharmaceutical formulations once they are able to form complexes with other molecules, improving their stability and bioavailability. The purpose of the present paper was to analyze complexes formed by papain/hydroxypropyl-beta-cyclodextrin and papain/beta-cyclodextrin by thermal analysis and cytotoxicity tests to verify their possible interactions and toxicological behavior. Complex solutions were prepared at different molar ratios and collected as a function of time to be lyophilized and analyzed. Results showed changes in endothermic events and cytotoxicity profiles. A complex formation for both complexes is observed; nevertheless, beta-cyclodextrin was able to change the enzyme characteristics more efficiently.

Transcriptional activation of MMP-13 by periodontal pathogenic LPS requires p38 MAP kinase

Matrix metal loprotease-13 (MMP-13) is induced by pro-inflammatory cytokines and increased expression is associated with a number of pathological conditions such as tumor metastasis, osteoarthritis, rheumatoid arthritis and periodontal diseases. MMP-13 gene regulation and the signal transduction pathways activated in response to bacterial LPS are largely unknown. In these studies, the role of the mitogen-activated protein kinase (MAPK) pathways in the regulation of MMP-13 induced by lipopolysaccharide was investigated. Lipopolysaccharide from Escherichia coli and Actinobacillus actinomycetemcomitans significantly (P < 0.05) increased MMP-13 steady-state mRNA (average of 27% and 46% increase, respectively) in murine periodontal ligament fibroblasts. MMP-13 mRNA induction was significantly reduced by inhibition of p38 MAP kinase. Immunoblot analysis indicated that p38 signaling was required for LPS-induced MMP-13 expression. Lipopolysaccharide induced proximal promoter reporter (-660/+32 mMMP-13) gene activity required p38 signaling. Collectively, these results indicate that lipopolysaccharide-induced murine MMP-13 is regulated by p38 signaling through a transcriptional mechanism.

Type 2 heat-labile enterotoxin (LT-II)-producing Escherichia coli isolated from ostriches with diarrhea

The culture supernatant of Escherichia coli, isolated from ostriches with diarrhea in Brazil, caused elongation in Vero cell, rounding in Chinese hamster ovary (CHO) cells and a cytoplasmic vacuolation in ostrich embryo fibroblasts (OEF), but it was not cytotoxic for chicken embryo fibroblasts (CEF). These effects were not neutralized by antiserum to cholera toxin. Polymerase chain reaction assays showed that the ostrich E.coli contained the gene encoding (eltII-A), but not those for type 1 heat-labile enterotoxin (eltA), heat-stable enterotoxins (estA, estB), verocytotoxins (stx-I, stx-II), or cytotoxic necrotizing factors (cnf 1, cnf 2). All isolates belonged to serotype O15:H8. The enteropathogenic relevance of LT-II in ostrich diarrhea remains undetermined. (C) 2004 Elsevier B.V. All rights reserved.

Paediatric rheumatology - A global perspective

This chapter aims to give a global perspective to paediatric rheumatology. The main points covered are the incidence, recognition of paediatric autoimmune diseases, and ethnic/geographic distribution. The most prevalent disease is juvenile idiopathic arthritis; robust data are still required for childhood-onset systemic lupus erythematosus, dermatomyositis, and scleroderma. Mimicking or overlapping infections are a major challenge in developing countries, and immunization policies in our patients in these areas need specific attention. The delivery of paediatric rheumatology care is also overviewed. Discrepancies in health-care resources and priorities are found in developing countries. Although most anti-rheumatic treatments are available worldwide, they are prohibitively expensive in many countries. For more traditional antirheumatic drugs there is still an ongoing need for good core outcome data across the world to ensure valid comparisons. Parent/patient education has been implemented worldwide in paediatric rheumatology through the power of the Internet. Physician and undergraduate training goals must be met to facilitate competent musculoskeletal assessment, a proper understanding of age-dependent variations, diagnosis, referral to specialists, and improved standards of care.

Porphyromonas gingivalis-mediated shedding of extracellular matrix metalloproteinase inducer (EMMPRIN) by oral epithelial cells: a potential role in inflammatory periodontal disease

Extracellular matrix metalloproteinase inducer (EMMPRIN) or CD 147 is a transmembrane glycoprotein expressed by various cell types, including oral epithelial cells. Recent studies have brought evidence that EMMPRIN plays a role in periodontitis. In the present study, we investigated the effect of Porphyromonas gingivalis, a major pathogen in chronic periodontitis, on the shedding of membrane-anchored EMMPRIN and on the expression of the EMMPRIN gene by oral epithelial cells. A potential contribution of shed EMMPRIN to the inflammatory process of periodontitis was analyzed by evaluating the effect of recombinant EMMPRIN on cytokine and matrix metalloproteinase (MMP) secretion by human gingival fibroblasts. ELISA and immunofluorescence analyses revealed that P. gingivalis mediated the shedding of epithelial cell-surface EMMPRIN in a dose- and time-dependent manner. Cysteine proteinase (gingipain)-deficient P. gingivalis mutants were used to demonstrate that both Arg- and Lys-gingipain activities are involved in EMMPRIN shedding. Real-time PCR showed that P. gingivalis had no significant effect on the expression of the EMMPRIN gene in epithelial cells. Recombinant EMMPRIN induced the secretion of IL-6 and MMP-3 by gingival fibroblasts, a phenomenon that appears to involve mitogen activated protein kinases. The present study brought to light a new mechanism by which P. gingivalis can promote the inflammatory response during periodontitis. (C) 2011 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Evaluation of the Effects of Endodontic Materials on Fibroblast Viability and Cytokine Production

Introduction: Recently, a new sealer composed of Portland cement named Endo-CPM-Sealer was developed. The aim of this study was to investigate the effects of Endo-CPM-Sealer (EGEO SRL, Buenos Aires, Argentina), Sealapex (Sybron Endo, Glendora, CA), and Angelus MTA (Angelus, Londrina, Brazil) on cell viability and cytokine (interleukin [IL]-1 beta and IL-6) production by mouse fibroblasts. Methods: Millipore culture plate inserts with polyethylene tubes filled with materials were placed into 24-well cell culture plates with mouse fibroblasts. Cells cultured with only empty polyethylene tubes were used as the control. After 24 hours, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay was used to evaluate the cell viability. For cytokine assay, mouse fibroblasts were incubated in 24-well flat-bottom plates with set material disks at the bottom. Cells cultured without the material disks served as the negative control. After 24 hours of incubation, culture media were collected for cytokine evaluation by using an enzyme-linked immunosorbent assay. The data were statistically analyzed by analysis of variance and Bonferroni correction. Results: Endo-CPM-Sealer, Sealapex, and Angelus MTA did not inhibit the cell viability. All materials induced IL-6 releasing, but the amount was not statistically significant compared with the control group. Angelus MTA induced IL-1 beta releasing significantly more than the control. Conclusions: All materials were not considered cytotoxic in fibroblast culture. (J Endod 2009;35:1577-1579)