Molecular data of UL24 homolog gene (ORF37) from Brazilian isolates of equine herpesvirus type 1

Equine herpesvirus type 1 (EHV-1) is associated with abortions, respiratory distress, and neurological disturbances in horses. The ORF37 of EHV-1 encodes a protein homolog to UL24 gene product of human herpesvirus that has been associated with neurovirulence. In the present work, ORF37 PCR fragments derived from two Brazilian EHV-1 isolates, a German isolate and an American reference strain were sequenced and characterized by molecular phylogenetic analysis. This genomic region is highly conserved an allowed to infer genetic distances between EHV-1 strains and other animal herpesvirus.     

Muscle reorganisation through local injection of stem cells in the diaphragm of mdx mice

Background

The diaphragm is the major respiratory muscle affected by Duchenne muscular dystrophy (DMD) and is responsible for causing 80% of deaths. The use of mechanical forces that act on the body or intermittent pressure on the airways improves the quality of life of patients but does not prevent the progression of respiratory failure. Thus, diseases that require tissue repair, such as DMD, represent a group of pathologies that have great potential for cell therapy. The application of stem cells directly into the diaphragm instead of systemic application can reduce cell migration to other affected areas and increase the chances of muscle reorganisation. The mdx mouse is a suitable animal model for this research because its diaphragmatic phenotype is similar to human DMD. Therefore, the aim of this study was to assess the potential cell implantation in the diaphragm muscle after the xenotransplantation of stem cells.

Methods

A total of 9 mice, including 3 control BALB/Cmice, 3 5-month-old mdx mice without stem cell injections and 3 mdx mice injected with stem cells, were used. The animals injected with stem cells underwent laparoscopy so that stem cells from GFP-labelled rabbit olfactory epithelium could be locally injected into the diaphragm muscle. After 8 days, all animals were euthanised, and the diaphragm muscle was dissected and subjected to histological and immunohistochemical analyses.

Results

Both the fresh diaphragm tissue and immunohistochemical analyses showed immunopositive GFP labelling of some of the cells and immunonegativity of myoblast bundles. In the histological analysis, we observed a reduction in the inflammatory infiltrate as well as the presence of a few peripheral nuclei and myoblast bundles.

Conclusion

We were able to implant stem cells into the diaphragm via local injection, which promoted moderate muscle reorganisation. The presence of myoblast bundles cannot be attributed to stem cell incorporation because there was no immunopositive labelling in this structure. It is believed that the formation of the bundles may have been stimulated by cellular signalling mechanisms that have not yet been elucidated.     

Update on the role of innate immune receptors during Brucella abortus infection

The innate immune system constitutes an efficient defense mechanism against invading microbial pathogens. Recent studies have revealed the intracellular signaling cascades involved in the TLR-initiated immune response to Brucella spp. infection. However, there is a piece of the puzzle missing that is the role of non-TLR receptors in innate immunity. The involvement of TLR receptors in brucellosis has been investigated by different research groups. It was demonstrated that TLR2 clearly does not play any role in controlling Brucella abortus infection in vivo, whereas TLR9 has been shown to be required for clearance of this bacterium in infected mice. The participation of adaptor molecules, such as MyD88 and TRIF has also been discussed. Recently, we and others have reported the critical role of MyD88- and not TRIF-mediated signaling in dendritic cell maturation and in vivo resistance during B. abortus infection. However, the relationship between specific Brucella molecules and non-TLR receptors and signal transduction pathways needs to be better understood. It is now clear that the interaction between TLRs and recently identified cytosolic innate immune sensors is crucial for mounting effective immune responses. Finally, this review discusses the mechanisms used by Brucella to escape detection by the host innate immune system.     

Learning by doing: a participatory methodology for systematization of experiments with agroforestry systems, with an example of its application

Participatory research methods have helped scientists to understand how farmers experiment and to seek partnerships with farmers in developing technologies with enhanced relevance and adoption. This paper reports on the development of a participatory methodology to systematize long-term experimentation with agroforestry systems carried out in a hotspot of biodiversity by non-governmental organizations and local farmers. A methodological guide for systematization and techniques used for Participatory Rural Appraisal formed the basis of our work. We propose an analytical framework that recognizes systems of reflexive and learning interactions, in order to make the learned lessons explicit. At the process level, the main lessons and recommendations are as follows. It is important to establish partnerships to conduct innovative and complex experimentation with agroforest. Participatory systematization allows us to improve the methodological aspects of design, implementation and management of on-farm participatory experimentation. It also serves to synthesize the main findings and to extract lessons from agroforestry systems experiments. It fosters the technical improvement of agroforestry systems. It creates possibilities for reflection on agroforestry systems by farmers, extensionists and researchers, as well as their learning with respect to management of such systems. The findings are placed in the context of current theory on participatory experimentation in agriculture. Extractive and interactive approaches help to produce rich insights of mutual interest through collaboration by identifying local, regional and global convergences, complementarities, and conflicts of interest; which affect the advance of new eco-friendly technologies, to both improve the livelihoods and to reverse biodiversity loss and environmental degradation.     

Extracellular matrix alterations in experimental Leishmania amazonensis infection in susceptible and resistant mice

ABSTRACT: Leishmania is inoculated, by the bite of an infected sandfly, into the skin of the host, where the promastigotes are phagocyted by dermal macrophages. The dermal region comprises cells and abundant extracellular matrix. Studies show that matrix metalloproteinases play an important role in host defense responses against pathogens in mammals and that their activities lead to the production of antimicrobial peptides. The aim of this study is to evaluate the changes in the distribution of fibronectin and laminin as well as in the elastic system fibres during the course of infection caused by Leishmania amazonensis in mice with distinct genetic backgrounds of susceptibility to this parasite. The results showed that BALB/c presented an enhancement of fibronectin during the course of infection when compared to their control group while the infected or non-infected C3H.He showed a decrease of this protein at end of the experiment. Laminin, on the other hand, remained unaltered in both strains. Also in both BALB/c and C3H.He mice the elastic and elaunin fibres remained unchanged while the oxytalan fibres decreased along the experiment. Ninety days after the infection C3H.He mice had recovered their capacity to produce oxytalan fibres.     

Hematological and coagulation profiles in dogs experimentally infected with Angiostrongylus vasorum (Baillet, 1866).

Hematological and coagulation profiles were studied in crossbred dogs experimentally infected with Angiostrongylus vasorum. Two groups of five dogs were experimentally inoculated with 50 and 100 third stage infective larvae (L(3)) of A. vasorum per kilogram of body weight. A third group of five uninfected animals was used as control. One sample of 10 ml of blood was collected from each animal on the 10, 20, 30, and 45 days after inoculation (dai) and at 30-day intervals thereafter for the remainder of the 210-day experimental period. The blood sample was used for the complete hemogram and platelet count, as well as measurements of prothrombin time, partial thromboplastin time and factors V and VIII. Anemia was observed in infected dogs, 6 weeks after the infection. The eosinophils presented peaks in four periods after infection. Thrombocytopenia became accentuated on the 72 dai. Decreased prothrombin time activity and increased partial thromboplastin time were observed at the 6 and 9 weeks after infection and decreased of factors VIII and V activities occurred from 4 to 6 weeks after infection. It may be conclude that infection by A. vasorum in dogs may cause a discrete anemia during the acute phase which is probably regenerative. In addition, important hemostatic alterations due to the infection suggest a chronic intravascular consumption coagulopathy.     

Determination of melamine content in amino resins by near-infrared spectroscopy

Melamine-urea-formaldehyde resin production has increased in the last years due to low formaldehyde emissions and improved moisture resistance when applied on wood-based panels. Melamine is a relatively high-cost raw material in the resin formulation and has a significant effect on wood-based panel properties. It is important for industry to implement efficient quality control methods to determine melamine content in resins. The main objective of this work was to develop chemometric techniques for off-line monitoring of melamine content using near-infrared spectroscopy. The use of interval partial least squares regression allowed identifying the most interesting spectral ranges for analysis (4,400–4,800 and 5,200–5,600 cm^?1) and the best preprocessing methods to use in the determination of this property. When the appropriate spectral range and preprocessing methods are selected, it is possible to obtain calibration models with high correlation coefficients.