Rutin promotes activation and reduces apoptosis of primordial follicles by regulating Akt phosphorylation after in vitro culture of ovine ovarian tissue.

The aims of this study were to analyze the effects of different concentrations of rutin on primordial follicle survival and development after in vitro culture of sheep ovarian tissue, and to verify the possible involvement of the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway in the rutin actions. Ovarian fragments were fixed for histological analysis (fresh control) or cultured in α-minimum essential medium alone (α-MEM+: control medium) or in α-MEM+supplemented with different concentrations of rutin (0.1; 1 or 10 µg/mL) for 7 days. Inhibition of the PI3K activity was performed in fragments cultured with 50 µM LY294002. Thereafter, immunohistochemistry was performed to evaluate the expression of cleaved caspase-3 (apoptosis) and Akt phosphorylation (p-Akt). The results showed that 1 µg/mL rutin has a greater percentage of normal follicles (P < 0.05) than those of α-MEM+ and other rutin treatments. In addition, 1 µg/mL rutin maintained the follicular apoptosis similar (P > 0.05) to that of the fresh control and lower than α-MEM+ and 10 µg/mL rutin. All rutin concentrations increased (P < 0.05) follicular activation compared to fresh control and α-MEM+. Furthermore, follicular and oocyte diameters increased (P < 0.05) only after culture with 1 µg/mL rutin. After PI3K inhibition, there was a reduction (P < 0.05) of rutin follicular effects. In conclusion, rutin at 1 µg/mL reduces apoptosis, promotes activation and growth of sheep primordial follicles through the modulation of the PI3K/Akt signaling pathway after in vitro culture of ovine ovarian tissue.

Assessment of Leishmania major and Leishmania braziliensis promastigote viability after photodynamic treatment with aluminum phthalocyanine tetrasulfonate (AlPcS4)

Cutaneous leishmaniasis is an infectious disease caused by protozoans of the genus Leishmania, which is transmitted through the bite of hematophagous insects of the genus Lutzomyia. This study aimed at testing in vitro the phototoxic effect of aluminum phthalocyanine tetrasulfonate (AlPcS4) on the viability of Leishmania major and Leishmania braziliensis. Stationary phase promastigote forms were treated with AlPcS4 at 1.0 µM and 10.0 µM and incubated for one hour. Then 659 nm laser was applied at 5 and 10 J/cm². Parasite viability was determined by differential count using the trypan blue dye exclusion method and by monitoring growth curves for nine days. Trypan blue exclusion assay showed a significant reduction of viable parasites compared to controls, L. major seemed more sensitive to the toxic effects of AlPcS4 in the dark. The most effective photodynamic therapy (PDT) was obtained with AlPcS4 at 10.0 µM and 10 J/cm² whereas L. braziliensis showed the highest mortality rate after treatment.

Immune response to Mycobacterium bovis-AN5 infection in genetically selected mice (selection IV-A)

Mice genetically selected for high (H) and low (L) antibody production (Selection IV-A) were used as murine experimental model. The aim of the present work was to evaluate the macrophagic activity and to characterize the immune response in Mycobacterium bovis-AN5 infected mice (3X107 bacteria). The response profile previously observed in such strains was not similar to that obtained during M. bovis infection; however, it corroborated works carried out using Selection I, which is very similar to Selection IV-A regarding infection by M. tuberculosis and Bacillus Calmette-Guérin (BCG). Considering bacterial recovery, LIV-A mice showed higher control of the infectious process in the lungs than in the spleen, whereas HIV-A mice presented more resistance in the spleen. With respect to macrophagic activity, hydrogen peroxide (H2O2) was probably not involved in the infection control since there was an inhibition in the production of this metabolite. Nitric oxide (NO) and TNF-a production seemed to be important in the control of bacterial replication and varied according to the strain, period and organ. Evaluation of the antibody production indicated that the multi-specific effect commonly observed in these strains was not the same in the response to M. bovis. Antibody concentrations were higher in LIV-A than in HIV-A mice at the beginning of the infection, being similar afterwards. Such data were compared with delayed-type hypersensitivity (DTH), which was more intense in HIV-A than in LIV-A mice, indicating that antibody production is independent of the capability to trigger DTH reactions and that cellular and humoral responses to M. bovis antigens show a polygenic control and an independent quantitative genetic regulation. Differences were observed among organs and metabolites, suggesting that different mechanisms play an important role in this infection in natural heterogeneous populations, indicating that NO, TNF-a and Th1 cytokines are involved in the infection control.(AU)

Cytokine production in lungs and adrenal glands of high and low antibody producing mice infected with Paracoccidioides brasiliensis.

Mice genetically selected for high (H) and low (L) antibody production (HIV-A and LIV-A) were used in an experimental model of paracoccidioidomycosis. In a previous work, it was observed that male HIV-A animals were more susceptible to the infection due to adrenal gland damage. Male HIV-A and LIV-A animals were intravenously inoculated with Paracoccidioides brasiliensis (strain 18) and sacrificed 2, 4, 6, 8 and 10 weeks after inoculation. At each time interval, lungs and adrenals were removed to estimate recoverability of the fungus, as well as to determine Th1 (IFN-gamma, TNF-alpha) and Th2 (IL-4 and IL-10) cytokine profiles. While viable fungi recoverability from the lungs of HIV-A mice was higher after 4 and 8 weeks, there was less fungal recovery from the adrenals of LIV-A animals after the 2nd week, with total fungal elimination after the 8th week. With regard to Th2 cytokines, there was an inhibition in IL-4 production in the organs from infected animals, the extent of which varied according to the organ and the time period after initiation of infection. IL-10 production was found to be lower in both organs. Determination of Th1 cytokines revealed that IFN-gamma production increased in both organs, mainly in the adrenal of LIV-A after 8 and 10 weeks, when these animals showed a total fungal elimination. A significant difference was observed between HIV-A and LIV-A concerning TNF-alpha production in both organs and at all recovery times, in that LIV-A produced a higher level of this cytokine, mainly in the adrenal. These results may explain the high susceptibility of HIV-A to P. brasiliensis infection, is due, at least in part, to adrenal involvement. The higher production of Th1 cytokines by LIV-A in comparison to HIV-A mice may account for LIV-A resistance to P. brasiliensis infection. Our data reveal the importance of this experimental model in the study of the adrenal involvement in paracoccidioidomycosis, since this gland may be highly compromised in the patients, leading to the development of Addison's Disease.

Efeito de diferentes doses de rutina sobre proteínas totais, enzimas aspartato amino trasferase e alanina amino transferase no soro de coelhos / Effect of different doses of rutin on total proteins, enzimes aspartate amino transferase and alanine amino transferase in serum of rabbits

Modelos com animais são usados para se pesquisar a toxicidade de fármacos, alimentos e novas substâncias com propriedades farmacológicas. Tem sido considerado que o consumo de flavonóides em alimentos é da ordem de 1g proveniente de vegetais, como frutas, leguminosas, etc. Com este objetivo, o presente trabalho avaliou os efeitos da rutina em diferentes doses, através das concentrações de proteínas, enzimas aspartato amino transferase (AST) e alanina amino transferase (ALT) em soro de coelhos. Alterações que ocorreram nos níveis séricos de proteína não foram estatisticamente significativos nos grupos e tempos avaliados para os machos e para as fêmeas. A atividade da AST também não representou diferença estatisticamente significativa. Já para ALT, observou-se variação apenas com 20mg de rutina para os machos nos tempos 0 e 28 dias. Através dos resultados obtidos, pode-se concluir que a rutina não causou patologias hepáticas.

[Hypocholesterolemic effect of naringin and rutin flavonoids]. / Efeito hipolipidêmico dos flavonóides naringina e rutina.

Flavonoids are pigments fenolics of plants that possess several biological activities, and many of these are associated with prevention of chronic diseases as cancer and hyperlipidemia. This work had as objective evaluates the effect of the flavonoids naringin and rutin on the metabolism lipidic of chicks hypercholesterolemic. In agreement with the results it can be observed that naringin and rutin reduced the levels of total cholesterol significantly, cholesterol-LDL, cholesterol-VLDL and triglycerols, not presenting, however, reductions in the levels of cholesterol-HDL.

Fatigue model to characterize cement-metal interface in dynamic shear.

This study evaluated the interface shear strength between vacuum mixed polymethyl-methacrylate and two types of surface enhancements under static and dynamic shear loading. Cobalt chrome coupons with grit blasted or light plasma sprayed surfaces were tested. For each test, two coupons were mounted in a stainless steel chamber such that they were bonded with a 2-mm cement mantle for each test. Pullout and dynamic fatigue tests were performed on an Instron machine. The mean static pullout strength of the grit blasted surfaces was 13.78 (+/- 2.73) MPa, whereas that of the light plasma sprayed surfaces was 18.46 (+/- 1.98) MPa. The rate of degradation of interface strength caused by fatigue was almost identical for both surface treatments. Qualitative analysis showed that the failed light plasma sprayed surface generated more metal and cement particles. These data suggest that light plasma sprayed surfaces sustain higher loads but have a potential for increased debris generation.

Polyethylene wear debris in modular acetabular prostheses.

The longevity of total hip arthroplasty has brought forth the recognition of aseptic loosening of prosthetic components as the leading cause of implant failure. Modularity of implants, although a significant improvement in versatility, may increase debris formation, a recognized cause of implant failure. This study was designed to measure the relative motion, and to assess the polyethylene wear debris production at the interface between the metal acetabular shell and the back side of the polyethylene liner, in modular hip prostheses. Five models from 4 manufacturers with different locking mechanisms and acetabular shell surface treatments were tested under long-term simultaneous sinusoidal and static loading (10(7) cycles at 3 Hz with +/- 2.5 Nmeter and 220 N static load). Results showed that there were marked differences in the security of the acetabular shell and polyethylene liner locking mechanism, wear pattern, damage sites, and amount of polyethylene debris on the acetabular shell and polyethylene liner surfaces. The range of polyethylene liner motion observed among the 5 models during 1 cycle of testing varied from an average of 0.96 degrees to movement too small to be detected by the test machines. Image and scanning electron microscopy analysis showed different wear patterns and a wide range in the average polyethylene liner surface wear area (0.26 cm2-4.61 cm2). In general, a stable locking mechanism and a smooth acetabular shell surface are essential in minimizing polyethylene liner wear and polyethylene debris production.

A method to characterize the passive elasticity in contracting muscle bundles.

There is concern among researchers whether the passive muscle properties, characterized by purely passive material testing procedures, are an appropriate representation of the actual passive component of the muscle. This aspect is of particular importance in the biomechanical analysis of heart muscle response where it is generally agreed that the so-called parallel elasticity cannot be ignored as is done justifiably in the analysis of skeletal muscle response. In the present article, a method of quantifying the passive elasticity in contracting muscle bundles is presented. The method consists of imposing isometric transients (such as the quick-stretch or quick release) on a muscle bundle during the contraction phase and observing the differences in decayed force levels between a normal twitch and that of a perturbed twitch. The proposed method provides a means of obtaining useful passive properties from contracting muscle bundles and circumvents the difficulty of having to characterize muscle properties from separate experiments on quiescent muscle bundles.

Isotropy and anisotropy of the arterial wall.

The passive biomechanical response of intact cylindrical rat carotid arteries is studied in vitro and compared with the mechanical response of rubber tubes. Using true stress and natural strain in the definition of the incremental modulus of elasticity, the tissue wall properties are analyzed over wide ranges of simultaneous circumferential and longitudinal deformations. The type of loading chosen is 'physiological' i.e. symmetric: the cylindrical segments are subjected to internal pressure and axial prestretch without torsion or shear. Several aspects pertaining to the choice of parameters characterizing the material are discussed and the analysis pertaining to the deformational behavior of a hypothetical compliant tube with Hookean wall material is presented. The experimental results show that while rubber response can be adequately represented as linearly elastic and isotropic, the overall response of vascular tissue is highly non-linear and anisotropic. However, for states of deformation that occur in vivo, the elasticity of arteries is quite similar to that of rubber tubes and as such the arterial wall may be viewed as incrementally isotropic for the range of deformations that occur in vivo.

A constitutive description of contracting papillary muscle and its implications to the dynamics of the intact heart.

A simple mathematical model capable of simulating the major biomechanical attributes of contracting cardiac muscle is presented. This model is based on the phenomenological observations on heart muscle. The form of the equation can be readily extended to describe the pressure-volume-time-velocity of the intact heart as well, thus allowing a direct bridge between the dynamics of papillary muscle and the dynamics of intact heart. Parameters that are sensitive to inotropic state of the muscle can be obtained directly from the isometric tension-time record of the muscle or the isovolumic pressure-time record of the ventricle. These parameters have the potential to serve as quantitative measures of cardiac health.

Visco-elasticity of passive cardiac muscle.

A quantitative mechanical description of the heart organ requires information on the mechanical behavior of its muscle in reasonable unity and completeness. In this respect, a fundamental constitutive law for soft biological tissures was proposed by Fung in 1972. This article presents evidence to show that Fung's law is a useful law to describe the mechanical behavior of heart muscle in the unstimulated (diastolic) state with sufficient generality. A visco-elastic relaxation phenomenon is studied in the isolated cardiac muscle of cat and rabbit with the purpose of constructing a mathematical model for relaxation. Experimental results show that passive relaxation behavior of heart muscle can be adequately described by a generalized standard linear solied with a continuous distribution of relaxation times. The form of the relaxation function devised permits the application of linear visco-elasticity theory to the nonlinear cardiac muscle. The relaxation model is used to predict the force-length (stress-strain) behavior of papillary muscle with reasonable accuracy.

Some mechanical considerations in the selection and testing of papillary muscles.

Measurements of segmental deformation made on papillary muscles obtained from cat, rabbit, dog and pig hearts suggest that the deformational behavior in these specimens is appreciably nonuniform both in the resting (passive) state and in the stimulated (active) state. In view of this, in the mechanical testing of papillary muscles, it is necessary to establish a minimum size of segment "sufficiently" far from the disturbing influence of end fixtures generally used to hold the specimen in the testing machine. The segment size should be large enough to average out the nonuniform aspects of deformation. Thus, the shape and size of the specimen dictated by the nonuniformities in the mechanical response, the thinness of the specimen dictated by the viability considerations and aspects of the testing machines and method dictated by the visco-elastic features of the specimen should be given due consideration in the selection and testing of papillary muscles.

Non-uniform strain distribution in papillary muscles.

Measurements of surface strains were made on small segments of intact papillary muscles of cats and rabbits. The measured strain is found to be non-uniform, both along the length and across the diameter of the papillary. This observation introduces difficulty in interpreting the data from experiments on the mechanics of the papillary muscle.

A device for testing mechanical properties of biological materials--the "Biodyne".

An electromechanical servo-controlled device has been developed. This device can be used to test the mechanical behavior of a wide variety of biological soft tissues. Control and execution of material testing procedures such as stress-strain, vibration, relaxation, creep etc. can be performed by manual operation of the device or by interfacing it with a laboratory type minicomputer. Experiments on excitable tissues such as muscle can also be executed. The design details and system performance are discussed.