Rhipicephalus (Boophilus) microplus (Canestrini, 1887) (Acari: Ixodidae): Acid phosphatase and ATPase activities localization in salivary glands of females during the feeding period

This study investigates the presence and the localization of acid phosphatase and ATPase in the salivary glands of Rhipicephalus (Boophilus) microplus female ticks during feeding. Semi-engorged females showed a larger amount of acid phosphatase compared to those at beginning of feeding, localized mainly in the apical portion of the secretory cells, and in the basal labyrinth of the interstitial cells. Ultrastructural observations also demonstrated its presence in secretion granules and inside some nuclei of secretory cells at beginning of feeding. Acid phosphatase in a free form probably has a hemolymph and/or ribosomal origin and participates in salivary gland secretion control. ATPase was detected in basal membrane of all types of acini and/or in the cytoplasm of the secretory cells at both feeding stages. The enzyme activities found strongly suggests that cell death by apoptosis occurs during the degenerative process. © 2006 Elsevier Inc. All rights reserved.

DNA damage and nitric oxide synthesis in experimentally infected Balb/c mice with Trypanosoma cruzi

This study aimed to evaluate whether experimental Chagas disease in acute phase under benznidazole therapy can cause DNA damage in peripheral blood, liver, heart, and spleen cells or induce nitric oxide synthesis in spleen cells. Twenty Balb/c mice were distributed into four groups: control (non-infected animals); Trypanosoma cruzi infected; T. cruzi infected and submitted to benznidazole therapy; and only treated with benznidazole. The results obtained with the single cell gel (comet) assay showed that T. cruzi was able induce DNA damage in heart cells of both benznidazole treated or untreated infected mice. Similarly, T. cruzi infected animals showed an increase of DNA lesions in spleen cells. Regarding nitric oxide synthesis, statistically significant differences (p < 0.05) were observed in all experimental groups compared to negative control, the strongest effect observed in the T. cruzi infected group. Taken together, these results indicate that T. cruzi may increase the level of DNA damage in mice heart and spleen cells. Probably, nitric oxide plays an important role in DNA damaging whereas benznidazole was able to minimize induced T. cruzi genotoxic effects in spleen cells. © 2006 Elsevier Inc. All rights reserved.

Taenia solium and Taenia saginata: Identification of sequence characterized amplified region (SCAR) markers

Cysticercosis is one of the most important zoonosis, not only because of the effects on animal health and its economic consequences, but also due to the serious danger it poses to humans. The two main parasites involved in the taeniasis-cysticercosis complex in Brazil are Taenia saginata and Taenia solium. Differentiating between these two parasites is important both for disease control and for epidemiological studies. The purpose of this work was to identify genetic markers that could be used to differentiate these parasites. Out of 120 oligonucleotide decamers tested in random amplified polymorphic DNA (RAPD) assays, 107 were shown to discriminate between the two species of Taenia. Twenty-one DNA fragments that were specific for each species of Taenia were chosen for DNA cloning and sequencing. Seven RAPD markers were converted into sequence characterized amplified region (SCAR) markers with two specific for T. saginata and five specific for T. solium as shown by agarose gel electrophoresis. These markers were developed as potential tools to differentiate T. solium from T. saginata in epidemiological studies. © 2007 Elsevier Inc. All rights reserved.

Performance of different genotypes of vegetable soybeans in Jaboticabal-SP

The study was conducted in the field, in the experimental area of the Section of Crop Production and Aromatic Medicinal Plants, belonging to the Faculty of Agricultural and Veterinary Sciences, Jaboticabal Campus-SP. The aim of the study was to assess the performance of seven genotypes of vegetable soybeans in Jaboticabal-SP. The experiment was carried out in a complete randomized block design with seven treatments (genotypes) and four replications. The genotypes examined were: CNPSOI, JLM003, JLM010, JLM018, JLM019, JLM024 and BRS216. The seeds were obtained from EMBRAPA-Soja and EMBRAPA- Hortaliças, and planted in Styrofoam trays with 128 cells containing Plantmax Hortaliças® as substrate. Transplanting occurred 10 d after seeding when the seedlings showed 2 or 3 definitive leaves and about 12 cm in height, demonstrating that the soil had been properly prepared according to the recommendations for this crop. Pests and diseases were adequately controlled in the event of their occurrence in the experimental area and in accordance with technical recommendations for the chemical products utilized. Collections were carried out based on the maturation of the pods, according to the scale of Fehr and Caviness (1977) adapted by Costa and Marchezan (1982), when the pods reached the reproductive stage R6. The parameters determined were mean precocity, height of the first pod, mean number of pods per plant, mean number of seeds per pod, fresh weight of 100 seeds and total yield of immature grains. Based on the results obtained, the genotypes JLM003 and JLM010 were found to be most indicated for growing vegetable soybeans, because of their capacity to produce immature grains of 1090 and 848 g·m-2, respectively, and fresh weights for 100 seeds of 62.20 and 68.19 g, respectively.

Mast cells modulate the inflammatory process in endotoxin-induced uveitis

Purpose: To investigate the role of mast cells and annexin-A1 (Anxa1) in endotoxin-induced uveitis (EIU). Methods: EIU was induced by injection of lipopolysaccharide (LPS) into the paws of rats, which were then sacrificed after 24 and 48 h. To assess EIU in the absence of mast cells, groups of animals were pretreated with compound 48/80 (c48/80) and sacrificed after 24 h after no treatment or EIU induction. The eyes were used for histological studies and the aqueous humor (AqH) pool was used for the analysis of transmigrated cells and Anxa1 levels. In inflammatory cells, Anxa1 expression was monitored by immunohistochemistry. Results: After 24 h, rats with EIU exhibited degranulated mast cells, associated with elevated numbers of infiltrating leukocytes and the high expression of Anxa1 in the AqH and the neutrophils. After 48 h of EIU, the mast cells were intact, indicating granule re-synthesis, and there was a reduction of neutrophil transmigration and an increase in the number of mononuclear phagocytic cells in ocular tissues. Anxa1 expression was decreased in neutrophils but increased in mononuclear phagocytic cells. In the animals pretreated with c48/80 and subjected to EIU, mast cells responded to this secretagogue by degranulating and few transmigrated neutrophils were observed. Conclustions: We report that mast cells are a potential source of pharmacological mediators that are strongly linked to the pathophysiology of EIU, and the endogenous protein Anxa1 is a mediator in the homeostasis of the inflammatory process with anti-migratory effects on leukocytes, which supports further studies of this protein as an innovative therapy for uveitis. © 2011 Molecular Vision.

Structure of a novel class II phospholipase D: Catalytic cleft is modified by a disulphide bridge

Phospholipases D (PLDs) are principally responsible for the local and systemic effects of Loxosceles envenomation including dermonecrosis and hemolysis. Despite their clinical relevance in loxoscelism, to date, only the SMase I from Loxosceles laeta, a class I member, has been structurally characterized. The crystal structure of a class II member from Loxosceles intermedia venom has been determined at 1.7. Å resolution. Structural comparison to the class I member showed that the presence of an additional disulphide bridge which links the catalytic loop to the flexible loop significantly changes the volume and shape of the catalytic cleft. An examination of the crystal structures of PLD homologues in the presence of low molecular weight compounds at their active sites suggests the existence of a ligand-dependent rotamer conformation of the highly conserved residue Trp230 (equivalent to Trp192 in the glycerophosphodiester phosphodiesterase from Thermus thermophofilus, PDB code: 1VD6) indicating its role in substrate binding in both enzymes. Sequence and structural analyses suggest that the reduced sphingomyelinase activity observed in some class IIb PLDs is probably due to point mutations which lead to a different substrate preference. © 2011 Elsevier Inc.

A survey of owner attitudes and experiences in managing diabetic dogs

A survey of the experiences of owners of diabetic dogs was carried out by distributing questionnaires to veterinary clinics in São Paulo, Brazil. A total of 93 dog owners were surveyed over a 4. month period and descriptive statistical analysis, frequency distribution and correlations were performed. In most cases the clinical condition of the dogs had improved following treatment and owners were largely satisfied with veterinary intervention. Injection of neutral protamine hagedorm twice daily was the most commonly prescribed treatment and respondents found this procedure easy to perform, although it did pose some difficulties when the owners were away from their animals. Cost was considered an important factor at the initiation of treatment programs. The information gleaned from this survey, in providing useful insights into the attitudes and experiences of owners of diabetic animals, will contribute to improvements in the treatment and management of such patients. © 2011 Elsevier Ltd.

Monotonicity and asymptotics of zeros of Laguerre-Sobolev-type orthogonal polynomials of higher order derivatives

In this paper we analyze the location of the zeros of polynomials orthogonal with respect to the inner product where α >-1, N ≥ 0, and j ∈ N. In particular, we focus our attention on their interlacing properties with respect to the zeros of Laguerre polynomials as well as on the monotonicity of each individual zero in terms of the mass N. Finally, we give necessary and sufficient conditions in terms of N in order for the least zero of any Laguerre-Sobolev-type orthogonal polynomial to be negative. © 2011 American Mathematical Society.

Structural insights into selectivity and cofactor binding in snake venom l-amino acid oxidases

l-Amino acid oxidases (LAAOs) are flavoenzymes that catalytically deaminate l-amino acids to corresponding α-keto acids with the concomitant production of ammonia (NH 3) and hydrogen peroxide (H 2O 2). Particularly, snake venom LAAOs have been attracted much attention due to their diverse clinical and biological effects, interfering on human coagulation factors and being cytotoxic against some pathogenic bacteria and Leishmania ssp. In this work, a new LAAO from Bothrops jararacussu venom (BjsuLAAO) was purified, functionally characterized and its structure determined by X-ray crystallography at 3.1å resolution. BjsuLAAO showed high catalytic specificity for aromatic and aliphatic large side-chain amino acids. Comparative structural analysis with prokaryotic LAAOs, which exhibit low specificity, indicates the importance of the active-site volume in modulating enzyme selectivity. Surprisingly, the flavin adenine dinucleotide (FAD) cofactor was found in a different orientation canonically described for both prokaryotic and eukaryotic LAAOs. In this new conformational state, the adenosyl group is flipped towards the 62-71 loop, being stabilized by several hydrogen-bond interactions, which is equally stable to the classical binding mode. © 2012 Elsevier Inc.

The involvement of anti-inflammatory protein, Annexin A1, in ocular toxoplasmosis

Purpose: The aim of this study was to evaluate the expression of the protein annexin A1 (ANXA1), a potent endogenous regulator of the inflammatory process, in ocular toxoplasmosis. Methods: C57BL/6 female mice were infected using intravitreal injections of either 10 6 tachyzoites of Toxoplasma gondii (RH strain; T. gondii) or PBS only (control groups). After 24, 48, and 72 h, animals were sacrificed and their eyes were harvested for histopathological, immunohistochemical, and ultrastructural immunocytochemical analysis of ANXA1. Human retinal pigment epithelial (RPE) cells (ARPE-19) were infected in vitro with T. gondii and collected after 60, 120, 240 min, and 24 h. Results: Compared with non-infected eyes, an intense inflammatory response was observed in the anterior (24 h after infection) and posterior segments (72 h after infection) of the infected eye, characterized by neutrophil infiltration and by the presence of tachyzoites and their consequent destruction along with disorganization of normal retina architecture and RPE vacuolization. T. gondii infection was associated with a significant increase of ANXA1 expression in the neutrophils at 24, 48, and 72 h, and in the RPE at 48 and 72 h. In vitro studies confirmed an upregulation of ANXA1 levels in RPE cells, after 60 and 120 min of infection with T. gondii. Conclusions: The positive modulation of endogenous ANXA1 in the inflammatory and RPE cells during T. gondii infection suggests that this protein may serve as a therapeutic target in ocular toxoplasmosis. © 2012 Molecular Vision.

Szego{double acute} and para-orthogonal polynomials on the real line: Zeros and canonical spectral transformations

We study polynomials which satisfy the same recurrence relation as the Szego{double acute} polynomials, however, with the restriction that the (reflection) coefficients in the recurrence are larger than one in modulus. Para-orthogonal polynomials that follow from these Szego{double acute} polynomials are also considered. With positive values for the reflection coefficients, zeros of the Szego{double acute} polynomials, para-orthogonal polynomials and associated quadrature rules are also studied. Finally, again with positive values for the reflection coefficients, interlacing properties of the Szego{double acute} polynomials and polynomials arising from canonical spectral transformations are obtained. © 2012 American Mathematical Society.

Fibronectin induces MMP2 expression in human prostate cancer cells

High-grade prostate cancers express high levels of matrix metalloproteinases (MMPs), major enzymes involved in tumor invasion and metastasis. However, the tumor cell lines commonly employed for prostate cancer research express only small amounts of MMPs when cultivated as monolayer cultures, in common culture media. The present study was conducted to ascertain whether culture conditions that include fibronectin can alter MMP2 and MMP9 expression by the human prostatic epithelial cell lines RWPE-1, LNCaP and PC-3. These cells were individually seeded at 2×104cells/cm2, cultivated until they reached 80% confluence, and then exposed for 4h to fibronectin, after which the conditioned medium was analyzed by gelatin zymography. Untreated cells were given common medium. Only RWPE-1 cells express detectable amounts of MMP9 when cultivated in common medium, whereas the addition of fibronectin induced high expression levels of pro and active forms of MMP2 in all tested cell lines. Our findings demonstrate that normal and tumor prostate cell lines express MMP2 activity when in contact with extracellular matrix components or blood plasma proteins such as fibronectin. Future studies of transcriptomes and proteomes in prostate cancer research using these cell lines should not neglect these important conclusions. © 2012 Elsevier Inc..

Kinematic analysis in healthy and hip-dysplastic German Shepherd dogs

This study investigated kinematic patterns in clinically normal German Shepherd dogs (GSDs) compared to those with hip dysplasia and with no clinical signs of lameness. Two groups of GSDs, including 10 clinically healthy dogs (G1) and 10 with hip dysplasia (G2), were trotted on a treadmill at a constant speed. Kinematic data were collected by a 3-camera system and analysed by a motion-analysis program. Flexion and extension joint angles and angular velocities were determined for the shoulder, elbow, carpal, hip, stifle, and tarsal joints.Within each group, the differences between the right and left limbs in all kinematic variables were not significant. Minimum angle, angular displacement and minimum angular velocity did not differ between groups. Significant differences were observed in the maximum angular velocity and maximum angle of the hip joint (dysplastic. >. healthy), and in the maximum angular velocity of the carpal joint (healthy. >. dysplastic). It was concluded that, when trotting on a treadmill, dysplastic dogs with no signs of lameness may present joint kinematic alterations in the hind as well as the forelimbs. © 2012 Elsevier Ltd.

Generic bifurcation of refracted systems

In this article we discuss some qualitative and geometric aspects of non-smooth dynamical systems theory. Our goal is to study the diagram bifurcation of typical singularities that occur generically in one parameter families of certain piecewise smooth vector fields named Refracted Systems. Such systems has a codimension-one submanifold as its discontinuity set. © 2012 Elsevier Ltd.

Toxicogenetic monitoring in urban cities exposed to different airborne contaminants

Microparticles found in the air may be associated with organic matter that contains several compounds, such as Polycyclic Aromatic Hydrocarbons (PAHs) and nitro-PAHs, and may pose a significant risk to human health, possibly leading to DNA mutations and cancers. This study associated genotoxicity assays for evaluating human exposure with the atmospheric air of two urban areas in southern Brazil, that received different atmospheric contributions. Site 1 was under urban-industrial influence and the other was a non-industrial reference, Site 2. Organic extracts from the airborne particulate matter were tested for mutagenicity via the Salmonella/microsome assay and analyzed for PAH composition. Cells samples of people residing in these two cities were evaluated using the comet and micronucleus assay (MN).Concentrations of the individual PAHs ranged from 0.01ng/m3 (benzo[a]anthracene) to 5.08ng/m3 (benzo[ghi]perylene). As to mutagenicity analysis of airborne, Site 1 presented all the mutagenic responses, which varied from 3.2±1.22rev/m3 (TA98 no S9) to 32.6±2.05rev/m3 (TA98, S9), while Site 2 ranged from negative to minimal responses. Site 1 presented a high quantity of nitro and amino derivatives of PAHs, and peaked at 56.0±3.68rev/μg (YG1024 strain). The two groups presented very low DNA damage levels without intergroup difference. Although Site 1 presented high mutagenic responses in the air samples, high PAH levels, healthy people exposed to this environment did not show significative damage in their genetic material. However, the evaluation of different environmental and genetic damage in such population is necessary to monitor possible damages. © 2013 Elsevier Inc.