Involvement of organic systems in golden retriever X-linked muscular dystrophy

Duchenne muscular dystrophy is a lethal genetic disease characterized by progressive muscle degeneration that usually had been used the Golden Retriever as a model for studying the disease (GRMD - Golden Retriever Muscular Dystrophy). A total of 16 male dystrophic Golden Retrievers dogs between 5 to 51 months of age were examined in the present study. The animals were classified as dystrophic according to two simultaneous complementary criteria: genotypic analysis and serum creatine kinase levels. The macroscopic abnormalities of the different organs and tissues and histopathological features were described using hematoxylin-eosin. The lesions in the skeletal muscles associated with the digestive problems resulted in cachexia with different intensities in all the dystrophic dogs. Cardiac muscle involvement was found in 87,5% of the GRMD dogs resulting, however, in cardiac failure in only 18,8% of the animals. The musculature of the diaphragm was hypertrophic in all affected animals resulting in progressive respiratory muscle weakness and at later stages in respiratory failure (81,25%). The liver abnormalities found in dystrophic dogs were originated mainly from heart disease and developed progressively. Hyperemia of mucosa and granular material indicated changes in the functioning and emptying of bladder. The germinative lineage cells presented moderate to severe degeneration probably due to degeneration of the scrotum and cremaster muscle which prevented the proper thermo-regulation of the testicle. Our results highlight the fact that there is significant impairment of the cardiac, respiratory and skeletal muscle systems in GRMD dogs since the age of five months. In addition, significant alterations of the gastrointestinal tract, urinary and reproductive systems are indicating the presence of degenerative lesions in the smooth musculature.

Influence of organic and mineral soil fertilization on essential oil of Spilanthes oleracea cv. Jambuarana

In the present study, the composition of essential oil of leaves and inflorescences of jambu (Spilanthes oleracea. Jambuarana), under organic manuring and mineral fertilization, was stuhed. Jambu plants show important chemical properties and their production has been addressed for the extraction of the essential oils for cosmetics industries, due to their pharmacolopcal properties. The experimental area of treatments contained urea as mineral fertilizer (120 g m2), applied twice and organic fertilizer (8 kg m2), applied at the planting. Jambu leaves and flowers were harvested twice: the first at 90 days after seedling transplantation and at the opening of the flower buds. Branches were cut at 7 cm from the soil, thus new branches can bud for the accomplishment of the second crop which happened 40 days after the re-budhng. The essential oil was analyzed by gas chromatography coupled with mass-spectrometry. According to our results the most representative compounds were trans-caryophyllene, germacrene-D, 1-dodecene, spathulenol and spilanthol (a compound presenting anesthetic properties) occurring in inflorescences. Fertilization procedure does not affect the content and the quality of the essential oil in Jambu plants. © 2012 Academic Journals Inc.

Evaluation of the effects of nicorandil and its molecular precursor (without radical NO) on proliferation and apoptosis of 786-cell

Nicorandil is a nitric oxide (NO) donor used in the treatment of angina symptoms. It has also been reported to protect cells and affect the proliferation and death of cells in some tissues. The molecules that interfere with these processes can cause dysfunction in healthy tissues but can also assist in the therapy of some disorders. In this study we examined the effect of nicorandil and of the molecular precursor that does not have the NO radical (N-(beta-hydroxyethyl) nicotinamide) on the cell proliferation and death of human renal carcinoma cells (786-O) under normal oxygenation conditions. The molecular precursor was used in order to analyze the effects independents of NO. In the cytotoxicity test, nicorandil was shown to be cytotoxic at very high concentrations and it was more cytotoxic than its precursor (cytotoxic at concentrations of 2,000 and 3,000 μg/mL, respectively). We propose that the lower cytotoxicity of the precursor is due to the absence of the NO radical. In this study, the cells exposed to nicorandil showed neither statistically significant changes in cell proliferation nor increases in apoptosis or genotoxicity. The precursor generated similar results to those of nicorandil. We conclude that nicorandil causes no changes in the proliferation or apoptosis of the cell 786-O in normal oxygenation conditions. Moreover, the lack of NO radical in the precursor molecule did not show a different result, except in the cell cytotoxicity. © 2013 Springer Science+Business Media Dordrecht.

Cellular responses in the Malpighian tubules of Scaptotrigona postica (Latreille, 1807) exposed to low doses of fipronil and boric acid

Studies of sub-lethal effects of pesticide residues on stingless bees are scarce and morphological analysis of organs would add information to toxicological analysis in order to clarify the continuous exposure of Scaptotrigona postica to insecticides. The aim of this study was to evaluate the morphology and histochemistry of the Malpighian tubules (excretory organ) of S. postica exposed to fipronil or boric acid to detect cellular responses that indicate toxicity or adaptative mechanisms to stress induced by exposure of worker bees to low doses of these chemical compounds. Newly emerged bees were submitted to toxicological bioassays and morphological analyses by optical microscopy and Transmission Electron Microscopy, as well as histochemical methods, were performed to detect proteins and glycoconjugates. Additionally, immunohistochemical detection of DNA fragmentation and HSP70 (70-kDa Heat shock protein) were performed to detect cell death and stress response, respectively. Statistical analysis, for the bioassays conducted with ingestion of contaminated diet with boric acid at 0.75% (w/w) or with fipronil at 0.1μg/kg of food, showed that the survival of bees that ingested the contaminated diets were significantly different to the survival rate presented by the control group (P<0.0001). Although some characteristics indicative of initiation of cell death were observed, the cells remained metabolically active in the processes of excretion and inactivation of chemical compounds. The data from this study reinforce the importance of research on sublethal effects of low doses of pesticides on bees in an attempt to assess a possible realistic dose and evaluate the risk assessment of stingless bee S. postica foraging in the vicinity of cultivated fields and/or in green urban areas. © 2013 Elsevier Ltd.

Indices of competition and bio-agroeconomic efficiency of lettuce and tomato intercrops in greenhouses

Intercropping is a cropping system for the production of greenhouse vegetables. It uses space more efficiently, thus reducing the cost of production. Intercropping tomato and lettuce has not been studied, but knowledge of the competitive and agroeconomic indices of these vegetables can help in the management of the intercropping system. The objectives of this study were to assess, through biological and agroeconomic indices, the competition between species and the profitability of intercropping tomato and lettuce at different times of transplantation over two growing seasons (autumn-winter and summer-winter) in greenhouse conditions. In autumn-winter, two experiments were conducted with a randomised complete-block design and five replicates. Tomato and lettuce were the main crops in the individual experiments. Treatments were arranged in a factorial of two cropping systems (intercropping and individual crops) with four transplants of the secondary crop (0, 10, 20 and 30 days after) plus an additional treatment (individual main crop). These two experiments were repeated in summer-winter. Tomato was the dominant crop regardless of transplant order. Intercropping systems established with transplants of both species on the same day had higher values of indices of competition and bio-agroeconomic efficiency than systems with longer periods of transplants between main and secondary crops. The intercropping of lettuce and tomato in greenhouses, regardless of transplant time or order, had bio-agroeconomic advantages over individual crops. The transplantation of tomato after lettuce is recommended for greater profitability.

SOCS3 expression correlates with severity of inflammation, expression of proinflammatory cytokines, and activation of STAT3 and p38 MAPK in LPS-induced inflammation in vivo

SOCS3 is an inducible endogenous negative regulator of JAK/STAT pathway, which is relevant in inflammatory conditions. We used a model of LPS-induced periodontal disease in rats to correlate SOCS3 expression with the inflammatory status. In vitro we used a murine macrophage cell line to assess the physical interaction between SOCS3 and STAT3 by coimmunoprecipitation. 30 ug of LPS from Escherichia coli were injected in the gingival tissues on the palatal aspect of first molars of the animals 3x/week for up to 4 weeks. Control animals were injected with the vehicle (PBS). The rats were sacrificed at 7, 15, and 30 days. Inflammation and gene expression were assessed by stereometric analysis, immunohistochemistry, RT-qPCR, and western blot. LPS injections increased inflammation, paralleled by an upregulation of SOCS3, of the proinflammatory cytokines IL-1β, IL-6, and TNF-and increased phosphorylation of STAT3 and p38 MAPK. SOCS3 expression accompanied the severity of inflammation and the expression of proinflammatory cytokines, as well as the activation status of STAT3 and p38 MAPK. LPS stimulation in a macrophage cell line in vitro induced transient STAT3 activation, which was inversely correlated with a dynamic physical interaction with SOCS3, suggesting that this may be a mechanism for SOCS3 regulatory function. © 2013 João Antônio Chaves de Souza et al.

Terapia de reposição enzimática para as mucopolissacaridoses I, II e VI: recomendações de um grupo de especialistas brasileiros

As mucopolissacaridoses (MPS) são doenças genéticas raras causadas pela deficiência de enzimas lisossômicas específicas que afetam o catabolismo de glicosaminoglicanos (GAG). O acúmulo de GAG em vários órgãos e tecidos nos pacientes afetados pelas MPS resulta em uma série de sinais e sintomas, integrantes de um quadro clínico multissistêmico que compromete ossos e articulações, vias respiratórias, sistema cardiovascular e muitos outros órgãos e tecidos, incluindo, em alguns casos, as funções cognitivas. Já foram identificados 11 defeitos enzimáticos que causam sete tipos diferentes de MPS. Antes do advento de terapias dirigidas para a restauração da atividade da enzima deficiente, o tratamento das MPS tinha como principal foco a prevenção e o cuidado das complicações, aspecto ainda bastante importante no manejo desses pacientes. Na década de 80 foi proposto o tratamento das MPS com transplante de medula óssea/transplante de células tronco hematopoiéticas (TMO/TCTH) e na década de 90 começou o desenvolvimento da Terapia de Reposição Enzimática (TRE), que se tornou uma realidade aprovada para uso clínico nas MPS I, II e VI na primeira década do século 21. Os autores deste trabalho têm a convicção de que um melhor futuro para os pacientes afetados pelas MPS depende da identificação, compreensão e manejo adequado das manifestações multissistêmicas dessas doenças, incluindo medidas de suporte (que devem fazer parte da assistência multidisciplinar regular destes pacientes) e terapias específicas. Embora a inibição da síntese de GAG e o resgate da atividade enzimática com moléculas pequenas também possam vir a ter um papel no manejo das MPS, o grande avanço disponível no momento é a TRE intravenosa. A TRE permitiu modificar radicalmente o panorama do tratamento das mucopolissacaridoses I, II e VI na última década, sendo que ainda pode estender seus benefícios em breve para a MPS IV A (cuja TRE já está em desenvolvimento clínico), com perspectivas para o tratamento da MPS III A e do déficit cognitivo na MPS II através de administração da enzima diretamente no sistema nervoso central (SNC). Um grande número de centros brasileiros, incluindo serviços de todas as regiões do país, já têm experiência com TRE para MPS I, II e VI. Essa experiência foi adquirida não só com o tratamento de pacientes como também com a participação de alguns grupos em ensaios clínicos envolvendo TRE para essas condições. Somados os três tipos de MPS, mais de 250 pacientes já foram tratados com TRE em nosso país. A experiência dos profissionais brasileiros, somada aos dados disponíveis na literatura internacional, permitiu elaborar este documento, produzido com o objetivo de reunir e harmonizar as informações disponíveis sobre o tratamento destas doenças graves e progressivas, mas que, felizmente, são hoje tratáveis, uma realidade que traz novas perspectivas para os pacientes brasileiros afetados por essas condições.

Immunodetection of Helicobacter sp. and the associated expression of ABO blood group antigens in the gastric mucosa of captive and free-living New World primates in the Amazon region

The histo-blood group ABH antigens were first described in humans. These antigens are only present on erythrocytes from great apes and humans, while in more primitive animals they are found in tissues and body fluids. The ABH antigens are mainly distributed in tissues exposed to the external environment and potentially serve as ligands for pathogens or inhibitors of tissue connections. The objective of this paper was two-fold: (i) to determine the presence of Helicobacter sp. in the gastric mucosa of 16 captive and 24 free-living New World monkeys and (ii) to evaluate the presence of histopathological alterations related to bacterial infection and the associated expression of ABH antigens in the tissue. Stomach tissues from 13 species of monkey were assessed using haematoxylin-eosin and modified Gram staining (Hucker) methods. An immunohistochemical analysis of the tissue revealed the presence of infectious bacteria that were characteristic of the genus Helicobacter sp. The results demonstrate that various species of monkey might be naturally infected with the Helicobacter sp. and that there is an increased susceptibility to infection. This study serves as a comparative analysis of infection between human and non-human primates and indicates the presence of a new species of Helicobacter.

Chronic graft-versus-host disease: clinical presentation of multiple lesions of lichenoid and atrophic pattern

Graft-versus-host disease is observed mainly in recipients of hematopoietic cell transplantation and is expressed by cutaneous or systemic signals and symptoms. Graft-versus-host disease is clinically classified as acute or chronic. Chronic Graft-versus-host disease occurs in up to 70% of hematopoietic cell transplanted patients and its clinical manifestations have important impact on morbidity and quality of life. The authors report an expressive cutaneous, oral and adnexal involvement in a patient with chronic Graft-versus-host disease with multiple lesions of lichenoid and atrophic pattern.

In Vitro Regulation of CCL3 and CXCL12 by Bacterial By-products Is Dependent on Site of Origin of Human Oral Fibroblasts

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

Histopathological Condition of the Remaining Tissues after Endodontic Infection of Rat Immature Teeth

Introduction: Recently, case reports have shown that immature teeth diagnosed with necrotic pulp and periapical periodontitis can be repaired through a regenerative endodontic procedure. True regeneration depends on the presence of stem cells in the remaining vital tissues. The aim of this study was to evaluate the histologic condition of the pulp tissue, root apical papilla, and periapical tissues after inducing endodontic infection in immature rat teeth for different periods. Methods: This study evaluated 18 first upper rat molars (36 roots). Periapical lesions were induced and were confirmed radiographically, and the animals were divided into 3 groups according to the days of pulp exposure for endodontic infection induction: 30, 60, and 90 days. Histologic analysis was performed in 5 different areas (ie, cervical, middle, and apical root canal thirds; the apical papilla; and the periapex surrounding the apical papilla). Results: At 30 days, one third of the specimens still showed vital but intensely inflamed pulp tissue in the apical third and vital apical papilla with varying degrees of inflammation. After 60 days, the results were similar with respect to the apical pulp tissue and apical papilla. Completely necrotic pulp tissue in the space canal and vital apical papilla were observed in about 67% of the cases after 90 days. Conclusions: Vital pulp tissue was observed in the apical third until 60 days and in the vital apical papilla until 90 days of infection in a rat model.

Microbiological quality of whole and filleted shelf-tilapia

Despite fish being a rich source of animal nutrients and having numerous associated health benefits, it is an extremely perishable food, prone to a wide range of hazards. The bacterial load associated with shelf-whole-fish organs (e.g. digestive tracts and skin) or mishandling of fish may be a vehicle of infection and become a risk to public health. The objective of this paper is to evaluate the microbiological quality of whole ungutted and filleted shelf-tilapia, as well as assess the safety for human consumption. For this purpose, in order to investigate the distribution and occurrence of bacterial populations, the count of total and thermotolerant coliforms, coagulase-positive Staphylococcus and presence of Salmonella spp. was determined. This paper shows that all fish organs were contaminated with thermotolerant coliform. Skin and fillet show higher populations and occurrence of all microorganisms analyzed. Lower bacterial populations were recovered from the gut and muscles of whole tilapia. Two samples of fillet were contaminated with coagulase-positive Staphylococcus. It can be concluded that the skin and filleted tilapia are important carriers of food-borne pathogens. In addition, fish might become an important cross and self-contamination source. (C) 2014 Elsevier B.V. All rights reserved.

Remodeling of cortical and corticocancellous fresh-frozen allogeneic block bone grafts - a radiographic and histomorphometric comparison to autologous bone grafts

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

Forced orthodontic eruption for augmentation of soft and hard tissue prior to implant placement

Forced orthodontic eruption (FOE) is a non-surgical treatment option that allows modifying the osseous and gingival topography. The aim of this article is to present a clinical case of a FOE, which resulted in an improvement of the amount of available bone and soft-tissues for implant site development. Patient was referred for treatment of mobility and unesthetic appearance of their maxillary incisors. Clinical and radiographic examination revealed inflamed gingival tissue, horizontal and vertical tooth mobility and interproximal angular bone defects. It was chosen a multidisciplinary treatment approach using FOE, tooth extraction, and immediate implant placement to achieve better esthetic results. The use of FOE, in periodontally compromised teeth, promoted the formation of a new bone and soft-tissue in a coronal direction, without additional surgical procedures, enabling an esthetic, and functional implant-supported restoration.

Construction of pediatric homogeneous phantoms for optimization of chest and skull radiographs

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)