Myofibroblasts in Schistosomal Portal Fibrosis of Man

Myofibroblasts, cells with intermediate features between smooth muscle cells and fibroblasts, have been described as an important cellular component of schistosomal portal fibrosis. The origin, distribution and fate of myofibroblasts were investigated by means of light, fluorescent, immunoenzymatic and ultrastructural techniques in wedge liver biopsies from 68 patients with the hepatosplenic form of schistosomiasis. Results demonstrated that the presence of myofibroblasts varied considerably from case to case and was always related to smooth muscle cell dispersion, which occurred around medium-sized damaged portal vein branches. By sequential observation of several cases, it was evident that myofibroblasts derived by differentiation of vascular smooth muscle and gradually tended to disappear, some of them further differentiating into fibroblasts. Thus, in schistosomal pipestem fibrosis myofibroblasts appear as transient cells, focally accumulated around damaged portal vein branches, and do not seem to have by themselves any important participation in the pathogenesis of hepatosplenic schistosomiasis.

Fibrogenesis and collagen resorption in the heart and skeletal muscle of Calomys callosus experimentally infected with Trypanosoma cruzi: immunohistochemical identification of extracellular matrix components

Intense inflammatory lesions and early development of interstitial fibrosis of the myocardium and skeletal muscle with spontaneous regression, have been described in Calomys callosus infected with Trypanosoma cruzi. The genetic types of collagen present in this model were investigated through immunohistochemistry using specific antibodies, combined with histopathology and Picro-Sirius staining of collagen. Thirty-five calomys were infected with the Colombian strain of T. cruzi and sacrificed at 24, 30, 40, 60 and 90 days post-infection. Inflammatory lesions and fibrogenesis were prominent at the early phase of infection and significantly decreased during late infection. Immunoisotyping of the matrix components was performed by indirect immunofluorescence on 5 µm thick cryostat sections using specific antibodies against laminin, fibronectin and isotypes I, III and IV of collagen. In the early phase, positive deposits of all the matrix components were present, with predominance of fibronectin, laminin and collagens types I and III in the myocardium and of types III and IV in the skeletal muscles. From the 40th day, type IV collagen predominates in the heart. At the late phase of infection (60th to 90th day), a clear fragmentation and decrease of all the matrix components were detected. Findings of the present study indicate that a modulation of the inflammatory process occurs in the model of C. callosus, leading to spontaneous regression of fibrosis independent of the genetic types of collagen involved in this process.

Intestinal fibrovascular nodules caused by Schistosoma mansoni infection in Calomys callosus Rengger, 1830 (Rodentia: Cricetidae): a model of concomitant fibrosis and angiogenesis

Human schistosomiasis develops extensive and dense fibrosis in portal space, together with congested new blood vessels. This study demonstrates that Calomys callosus infected with Schistosoma mansoni also develops fibrovascular lesions, which are found in intestinal subserosa. Animals were percutaneously infected with 70 cercariae and necropsied at 42, 45, 55, 80, 90 and 160 days after infection. Intestinal sections were stained for brightfield, polarization microscopy, confocal laser scanning, transmission and scanning electron microscopies. Immunohistological analysis was also performed and some nodules were aseptically collected for cell culture. Numerous intestinal nodules, appearing from 55 up to 160 days after infection, were localized at the interface between external muscular layer and intestinal serosa, consisting of fibrovascular tissue forming a shell about central granuloma(s). Intranodular new vessels were derived from the vasculature of the external vascular layer and were positive for laminin, chondroitin-sulfate, smooth muscle alpha-actin and FVIII-RA. Fibroblastic cells and extracellular matrix components (collagens I, III and VI, fibronectin and tenascin) comprised the stroma. Intermixed with the fibroblasts and vessels there were variable number of eosinophils, macrophages and haemorrhagic foci. In conclusion, the nodules constitute an excellent and accessible model to study fibrogenesis and angiogenesis, dependent on S. mansoni eggs. The fibrogenic activity is fibroblastic and not myofibroblastic-dependent. The angiogenesis is so prominent that causes haemorrhagic ascites.

Schistosomal hepatopathy

Gross anatomical features and a complex set of vascular changes characterize schistosomal hepatopathy as a peculiar form of chronic liver disease, clinically known as "hepatosplenic schistosomiasis". It differs from hepatic cirrhosis, although clinical and pathological aspects may sometimes induce confusion between these two conditions. Intrahepatic portal vein obstruction and compensatory arterial hypertrophy render the hepatic parenchyma vulnerable to ischemic insult. This may lead to focal necrosis, which may give place to focal post-necrotic scars. These events are of paramount importance for the clinico-pathological evolution of schistosomal hepatopathy. Although portal fibrosis due to schistosomiasis sometimes reveals numerous myofibroblasts, it does not mean that such fibrosis belongs to a peculiar type. Damage to the muscular walls of the portal vein may be followed by dissociation of smooth muscle cells and their transition toward myofibroblasts, which appear only as transient cells in schistosomal portal fibrosis. Studies made with plastic vascular casts, especially those with the murine model of "pipestem" fibrosis have helped to reveal the mechanisms involved in systematized portal fibrosis formation. However, the factors involved in the pathogenesis of hepatosplenic disease remain poorly understood. A process of chronic hepatitis is a common accompaniment of portal fibrosis in schistosomiasis. Most of the times it is caused by concomitant viral infection. However, no especial interaction seems to exist between schistosomal hepatopathy and viral hepatitis.

P2X1 receptors and the endothelium

Adenosine triphosphate (ATP) is now established as a principle vaso-active mediator in the vasculature. Its actions on arteries are complex, and are mediated by the P2X and P2Y receptor families. It is generally accepted that ATP induces a bi-phasic response in arteries, inducing contraction via the P2X and P2Y receptors on the smooth muscle cells, and vasodilation via the actions of P2Y receptors located on the endothelium. However, a number of recent studies have placed P2X1 receptors on the endothelium of some arteries. The use of a specific P2X1 receptor ligand, a, b methylene ATP has demonstrated that P2X1 receptors also have a bi-functional role. The actions of ATP on P2X1 receptors is therefore dependant on its location, inducing contraction when located on the smooth muscle cells, and dilation when expressed on the endothelium, comparable to that of P2Y receptors.

Interaction and cystogenesis of Toxoplasma gondii within skeletal muscle cells in vitro

Infection by the protozoan parasite Toxoplasma gondii is widely prevalent in humans and animals. To prevent human infection, all meat should be well cooked before consumption, since the parasite is present in skeletal muscle. In this context, the use of skeletal muscle cells (SkMCs) as a cellular model opens up new approaches to investigate T. gondii-host cell interactions. Immunofluorescent detection of proteins that are stage-specific for bradyzoites indicated that complete cystogenesis of T. gondii in in vitro cultures of SkMCs occurs after 96 h of infection. Ultrastructural analysis showed that, after 48 h of interaction, there were alterations on the parasitophorous vacuole membrane, including greater thickness and increased electron density at the inner face of the membrane. The present study demonstrates the potential use of primary cultures of SkMCs to evaluate different molecular aspects of T. gondii invasion and cystogenesis and presents a promising in vitro model for the screening of drug activities toward tissue cysts and bradyzoites.

Spontaneous stage differentiation of mouse-virulent Toxoplasma gondii RH parasites in skeletal muscle cells: an ultrastructural evaluation

Although the predilection for Toxoplasma gondii to form cysts in the nervous system and skeletal and heart muscles has been described for more than fifty years, skeletal muscle cells (SkMCs) have not been explored as a host cell type to study the Toxoplasma-host cell interaction and investigate the intracellular development of the parasite. Morphological aspects of the initial events in the Toxoplasma-SkMC interaction were analysed and suggest that there are different processes of protozoan adhesion and invasion and of the subsequent fate of the parasite inside the parasitophorous vacuole (PV). Using scanning electron microscopy,Toxoplasma tachyzoites from the mouse-virulent RH strain were found to be attached to SkMCs by the anterior or posterior region of the body, with or without expansion of the SkMC membrane. This suggests that different types of parasite internalization occurred. Asynchronous multiplication and differentiation of T. gondii were observed. Importantly, intracellular parasites were seen to display high amounts of amylopectin granules in their cytoplasm, indicating that tachyzoites of the RH strain were able to differentiate spontaneously into bradyzoites in SkMCs. This stage conversion occurred in approximately 3% of the PVs. This is particularly intriguing as tachyzoites of virulent Toxoplasma strains are not thought to be prone to cyst formation. We discuss whether biological differences in host cells are crucial to Toxoplasma stage conversion and suggest that important questions concerning the host cell type and its relevance in Toxoplasma differentiation are still unanswered.

Trichomonas vaginalis and Tritrichomonas foetus: interaction with fibroblasts and muscle cells - new insights into parasite-mediated host cell cytotoxicity

Trichomonas vaginalis and Tritrichomonas foetus are parasitic, flagellated protists that inhabit the urogenital tract of humans and bovines, respectively. T. vaginalis causes the most prevalent non-viral sexually transmitted disease worldwide and has been associated with an increased risk for human immunodeficiency virus-1 infection in humans. Infections by T. foetus cause significant losses to the beef industry worldwide due to infertility and spontaneous abortion in cows. Several studies have shown a close association between trichomonads and the epithelium of the urogenital tract. However, little is known concerning the interaction of trichomonads with cells from deeper tissues, such as fibroblasts and muscle cells. Published parasite-host cell interaction studies have reported contradictory results regarding the ability of T. foetus and T. vaginalis to interact with and damage cells of different tissues. In this study, parasite-host cell interactions were examined by culturing primary human fibroblasts obtained from abdominal biopsies performed during plastic surgeries with trichomonads. In addition, mouse 3T3 fibroblasts, primary chick embryo myogenic cells and L6 muscle cells were also used as models of target cells. The parasite-host cell cultures were processed for scanning and transmission electron microscopy and were tested for cell viability and cell death. JC-1 staining, which measures mitochondrial membrane potential, was used to determine whether the parasites induced target cell damage. Terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling staining was used as an indicator of chromatin damage. The colorimetric crystal violet assay was performed to ana-lyse the cytotoxicity induced by the parasite. The results showed that T. foetus and T. vaginalis adhered to and were cytotoxic to both fibroblasts and muscle cells, indicating that trichomonas infection of the connective and muscle tissues is likely to occur; such infections could cause serious risks to the infected host.

Urinary incontinence related to perineal muscle strength in the first trimester of pregnancy: cross-sectional study

Objective To analyze pelvic floor muscle strength (PFMS), urinary continence and quality of life related to urinary incontinence (UI) of women in the first trimester of pregnancy. Method Cross-sectional study with a sample of 500 women who started prenatal care in a complementary healthcare facility in Guarulhos, state of São Paulo, from 2012 and 2013. Pelvic floor muscle strength was evaluated through perineometry. The pregnant women who presented UI answered the International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF). Results It was found that maternal age (OR=1.06; CI95% 1.02-1.11) and prior UI (OR=15.12; 95%CI 8.19-27.92) are the variables that, in tandem, best explain the occurrence of UI at the beginning of pregnancy. The mean score on the ICIQ-SF was 8.2 (SD=3.9), considered a moderate impact on quality of life. Conclusion Older pregnant women with prior UI are more likely to have UI in the first trimester of pregnancy.


Dietary effects on muscle fatty acid composition of finished heifers

The effects of diet on Longissimus muscle fatty acid composition was determined using 24 crossbred heifers of Simmental vs. Nelore and Limousin vs. Nelore. The experimental diets were: 1) corn and yeast (CY); 2) corn, cottonseed meal + meat and bones meal (CMB); 3) cassava hull and yeast (CHY); 4) cassava hull, cottonseed meal + meat and bones meal (CHMB). Feeding CHMB diets resulted in lower lipid and higher cholesterol contents (P<0.05) for both crosses. Most of the identified fatty acids were monounsaturated, and the highest percentage was found to oleic acid (C18:1w9), with values ranging from 32.54 to 46.42%. Among the saturated fatty acids the palmitic acid (C16:0) showed the highest percentage, with its contents ranging between 19.40 and 32.44%. The highest polyunsaturated/saturated fatty acid ratio was of 0.30, and the lowest was of 0.08. Feeding CY diets resulted in lower cholesterol and higher polyunsaturated fatty acid contents of the Longissimus muscle.

Salting kinetics and salt diffusivities in farmed Pantanal caiman muscle

The legal Pantanal caiman (Caiman crocodilus yacare) farming, in Brazil, has been stimulated and among meat preservation techniques the salting process is a relatively simple and low-cost method. The objective of this work was to study the sodium chloride diffusion kinetics in farmed caiman muscle during salting. Limited volumes of brine were employed, with salting essays carried at 3, 4 and 5 brine/muscle ratios, at 15%, 20% and 25% w/w brine concentrations, and brine temperatures of 10, 15 and 20ºC. The analytical solution of second Fick's law considering one-dimensional diffusion through an infinite slab in contact with a well-stirred solution of limited volume was used to calculate effective salt diffusion coefficients and to predict the sodium chloride content in the fillets. A good agreement was obtained between the considered analytical model and experimental data. Salt diffusivities in fillets were found to be in the range of 0.47x10-10 to 9.62x10-10 m²/s.

Micropropagação do híbrido PExSC-52 e da cultivar Smooth Cayenne de abacaxizeiro

A produção de mudas de abacaxizeiro de qualidade, em curto período, exige a otimização de protocolos de multiplicação in vitro. O objetivo deste trabalho foi testar cinco combinações de fitorreguladores, baseadas em resultados de pesquisa publicados com micropropagação de abacaxizeiro, na multiplicação in vitro do híbrido PExSC-52 e da cultivar Smooth Cayenne. Gemas axilares foram cultivadas em meio MS contendo benzilaminopurina (BAP) 2 mg/L; BAP 2 mg/L + ácido naftaleno acético (ANA) 2 mg/L; cinetina (CIN) 2 mg/L; CIN 2 mg/L + ANA 2 mg/L e CIN 5 mg/L + ácido indolacético (AIA) 5 mg/L. No híbrido, BAP 2 mg/L proporcionou maior proliferação de brotos por explante por subcultura e taxa de multiplicação crescente até a quarta subcultura. BAP 2 mg/L + ANA 2 mg/L multiplicou melhor a cv. Smooth Cayenne. Quando se utilizou CIN praticamente não ocorreu proliferação de brotos. Brotações do híbrido provenientes da quarta subcultura em presença de BAP 2 mg/L e BAP 2 mg/L + ANA 2 mg/L foram enraizados em meio MS contendo: ANA 1 mg/L; ANA 1 mg/L + CIN 2 mg/L; ANA 0,5 mg/L + CIN 2 mg/L; CIN 2 mg/L e sem fitorregulador. Brotos multiplicados em BAP enraizaram melhor que aqueles multiplicados em BAP + ANA.

Fenologia e escalonamento da produção do abacaxizeiro 'Smooth Cayenne' no cerrado de Mato Grosso

O objetivo deste trabalho foi avaliar os efeitos da época de plantio e da idade de indução floral sobre a fenologia, o período de colheita e o escalonamento da produção do abacaxizeiro 'Smooth Cayenne', no Cerrado de Mato Grosso. O experimento foi conduzido em Tangará da Serra, MT, de novembro de 2005 a setembro de 2008. Foram testadas seis épocas de plantio - janeiro, março, maio, julho, setembro, novembro - e cinco idades para indução floral - 8, 10, 12, 14, 16 meses após o plantio -, além da floração natural. As variáveis analisadas foram: períodos entre indução floral e floração, floração e colheita, indução e colheita, e plantio e colheita; massas da coroa e do fruto com coroa; e comprimento da folha D. Não houve interação entre as épocas de plantio e de indução floral. A indução floral realizada nos meses de maio e julho, período de baixa temperatura e precipitação, alongou o ciclo da planta e diminuiu a massa dos frutos e o comprimento da folha D. A indução realizada em setembro e novembro determinou a colheita em épocas favoráveis de comercialização, independentemente do mês de plantio, sem interferência na massa dos frutos.

ALTAS DENSIDADES DE PLANTIO NA CULTURA DO ABACAXI CV. SMOOTH CAYENNE, SOB CONDIÇÕES DE SEQUEIRO

No Brasil têm-se usado densidades de plantio relativamente baixas para a obtenção de frutos grandes, com reflexos negativos na produtividade da cultura do abacaxi. No entanto, frutos cada vez menores têm sido comercializados no mercado internacional, o que poderá também ocorrer no mercado nacional. Este trabalho teve o objetivo de avaliar o efeito de altas densidades em sistemas de plantio em filas duplas sobre a produção quantitativa e qualitativa de abacaxi cv. Smooth Cayenne, sob condições de sequeiro. Densidades de plantio, variando de 51.280 plantas/ha a 100.000 plantas/ha, foram estudadas em delineamento experimental de blocos casualizados, com cinco repetições e doze tratamentos, distribuídos em esquema fatorial 4 x 3, correspondendo a quatro combinações de espaçamentos entre linhas duplas e entre linhas simples na fila dupla (90cm x 40 cm, 90 cm x 30 cm, 80 cm x 30 cm , 70 cm x 30 cm) e três espaçamentos entre plantas nas linhas de plantio (30 cm, 25 cm, 20 cm). A análise de variância determinou diferença estatística apenas para a produtividade, em função do espaçamento entre plantas na linha de plantio, sendo mais elevada para o espaçamento de 20 cm. O peso do fruto, as suas dimensões e a sua qualidade (açucares, acidez, teor de suco, relação açúcares/acidez) não foram significativamente influenciados pelas densidades de plantio estudadas, mantendo-se dentro dos padrões da cultivar. Para cada aumento de 10.000 plantas por hectare, a produtividade cresceu em 8,27 t/ha e o peso médio do fruto caiu 102 g. Nas condições ambientais dos Tabuleiros Costeiros do Norte da Bahia, a cultura do abacaxi cv. Smooth Cayenne apresenta potencial para uso em altas densidades de plantio, mesmo em cultivo de sequeiro, podendo-se atingir produtividade acima de 80t/ha e peso médio do fruto superior a 1,0 kg.

AVALIAÇÃO DE PLANTAS MATRIZES DE ABACAXIZEIRO CULTIVAR SMOOTH CAYENNE UTILIZANDO MARCADORES RAPD E PADRÕES ISOENZIMÁTICOS

Foram coletadas, em área comercial da fazenda Córrego dos Bois, município de Canápolis -- MG, 20 plantas matrizes de abacaxizeiro cultivar Smooth Cayenne, para avaliação de similaridade e padrões genotípicos através de marcadores moleculares RAPD e padrões isoenzimáticos. As plantas matrizes foram selecionadas mediante as seguintes características: planta sadia, frutos cilíndricos, ausência de fasciação, pedúnculo curto, ausência de espinhos nas folhas, "olho" do fruto chato e peso dos frutos entre 1,6 a 2,2kg. Para análise de marcadores RAPD, foram testados 100 "primers", dos quais, 43 foram eficientes na amplificação das amostras, onde foram observados padrões de bandas diferentes entre as plantas matrizes utilizadas, indicando a existência de variabilidade genética. Nos padrões isoenzimáticos, dos 15 sistemas utilizados para revelação das amostras, 8 apresentaram atividade enzimática, sendo 5 deles com baixa resolução; entretanto, estes sistemas não foram eficientes em diferenciar as amostras devido à ausência de polimorfismo