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Combined immunization using DNA-Sm14 and DNA-Hsp65 increases CD8+ memory T cells, reduces chronic pathology and decreases egg viability during Schistosoma mansoniinfection
No full textAbstract
Background
Schistosomiasis is one of the most important neglected diseases found in developing countries and affects 249 million people worldwide. The development of an efficient vaccination strategy is essential for the control of this disease. Previous work showed partial protection induced by DNA-Sm14 against Schistosoma mansoni infection, whereas DNA-Hsp65 showed immunostimulatory properties against infectious diseases, autoimmune diseases, cancer and antifibrotic properties in an egg-induced granuloma model.
Methods
C57BL/6 mice received 4 doses of DNA-Sm14 (100 μg/dose) and DNA-Hsp65 (100 μg/dose), simultaneously administrated, or DNA-Sm14 alone, once a week, during four weeks. Three groups were included: 1- Control (no immunization); 2- DNA-Sm14; 3- DNA-Sm14/DNA-Hsp65. Two weeks following last immunization, animals were challenged subcutaneously with 30 cercariae. Fifteen, 48 and 69 days after infection splenocytes were collected to evaluate the number of CD8+ memory T cells (CD44highCD62low) using flow cytometry. Forty-eight days after challenge adult worms were collected by portal veins perfusion and intestines were collected to analyze the intestinal egg viability. Histological, immunohistochemical and soluble quantification of collagen and α-SMA accumulation were performed on the liver.
Results
In the current work, we tested a new vaccination strategy using DNA-Sm14 with DNA-Hsp65 to potentiate the protection against schistosomiasis. Combined vaccination increased the number of CD8+ memory T cells and decreased egg viability on the intestinal wall of infected mice. In addition, simultaneous vaccination with DNA-Sm14/DNA-Hsp65 reduced collagen and α-SMA accumulation during the chronic phase of granuloma formation.
Conclusion
Simultaneous vaccination with DNA-Sm14/DNA-Hsp65 showed an immunostimulatory potential and antifibrotic property that is associated with the reduction of tissue damage on Schistosoma mansoni experimental infection
Microstructural analysis in the Fe-30.5Mn-8.0Al-1.3C and Fe-30.5Mn-2.1Al.1.2C steels upon cold rolling
No full textElectron backscatter diffraction (EBSD) and electron channeling contrast imaging (ECCI) were used to examine microstructural changes of the austenitic low-density Fe-30.5Mn-8.0Al-1.2C (8Al) and Fe-30.5Mn-2.1Al-1.2C (2Al) (wt.%) steels during cold rolling. As the strain increased, deformation mechanisms, such as stacking faults, slip, mechanical twinning, and shear banding were activated in both steels cold rolled up to strain of 0.69. Only slip was noted in these steels at low strain (ε=0.11) and slip dominance was detected in the 8Al steel at higher strains. Shear banding became active at higher strain (ε~0.7) in these materials. An inhomogeneous microstructure formed in both alloys at such strain level. More extensive mechanical twinning in the 2Al alloy than that in the 8Al alloy was observed. Fish bone-like structure patterns were revealed in the 8Al steel and a river-like structure in the 2Al steel. Detailed microstructure features as elongated and fragmented grains along the rolling direction (RD) were found for both steels, as already observed in other high-Mn steels. These deformed structures are composed by lamellar packets which can contain mechanical twins or slip lines and shear bands.CNP
High-fat diet or low-protein diet changes peritoneal macrophages function in mice
No full textAbstract\ud
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Background\ud
Obesity and protein malnutrition are major food problems nowadays, affecting billions of people around the world. The nutrition transition that has occurred in recent decades is changing the nutritional profile, reducing malnutrition and increasing the percentage of obese people. The innate immune response is greatly influenced by diet, with significant changes in both malnutrition and obesity. Therefore, we investigate the effects of protein malnutrition and obesity in nutritional and immunological parameters in mice.\ud
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Results\ud
Peritoneal macrophages of malnourished animals showed reduced functions of adhesion, spreading, and fungicidal activity; production of hydrogen peroxide and nitric oxide were lower, reflecting changes in the innate immune response. However, the high-fat animals had macrophage functions slightly increased.\ud
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Conclusions\ud
Animals subjected to low-protein diet have immunosuppression, and animals subjected to high-fat diet increased visceral adipose tissue and the presence of an inflammatory process with increased peritoneal macrophage activity and similar systemic changes to metabolic syndrome.The authors thank E. Makiyama and M. C. Ferreira for the technical\ud
assistance.This work was financially supported by the Fundação de Amparo a\ud
Pesquisa do Estado de São Paulo (FAPESP 2012/24505-1). E.W.S. was\ud
supported by a fellowship from CNPq (152770/2011-9)
Characterization and methodology for calculating the mechanical properties of a TRIP-steel submitted to hot stamping and quenching and partitioning (Q&P)
No full textThermomechanical simulation of quenching, hot stamping, and quenching and partitioning processes of a high-strength TRIP-assisted steel were carried out in a Gleeble®3S50 thermo-mechanical simulator, coupled to the synchrotron X-ray diffraction line. The microstructures and mechanical properties were analyzed using Field Emission Gun Scanning Electron Microscopy (FEG-SEM), X-ray diffraction, and nanoindentation. The microstructures of thermomechanical treated specimens were modeled using the Object Oriented Finite Element (OOF) technique. The modeled microstructures were then fed into a finite element model to predict the mechanical behavior. By using a reverse algorithm method, the elasto-plastic mechanical properties of different microconstituents were determined. This was done through the analysis of instrumented nanoindentation loading-penetration curves. Tensile properties of the thermomechanical processed steels were measured by tensile testing of subsized specimens cut from samples processed on the Gleeble®3S50. The comparison between the experimental results and those of the reverse algorithm and the OOF modeled microstructure showed quite good agreement.CAPESFAPESPLNNan
Transcriptional profiling reveals intrinsic mRNA alterations in multipotent mesenchymal stromal cells isolated from bone marrow of newly-diagnosed type 1 diabetes patients
No full textAbstract\ud
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Background\ud
Bone marrow multipotent mesenchymal stromal cells (MSCs) are a diverse subset of precursors that contribute to the homeostasis of the hematopoietic niche. MSCs can be isolated and expanded in vitro and have unique immunomodulatory and regenerative properties that make them attractive for the treatment of autoimmune diseases, including type 1 diabetes (T1D). Whether autologous or allogeneic MSCs are more suitable for therapeutic purposes has not yet been established. While autologous MSCs may present abnormal function, allogeneic cells may be recognized and rejected by the host immune system. Thus, studies that investigate biological characteristics of MSCs isolated from T1D patients are essential to guide future clinical applications.\ud
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Methods\ud
Bone marrow-derived MSCs from recently diagnosed type 1 diabetes patients (T1D-MSCs) were compared with those from healthy individuals (C-MSCs) for morphological and immunophenotypic characteristics and for differentiation potential. Bioinformatics approaches allowed us to match absolute and differential gene expression of several adhesion molecules, immune mediators, growth factors, and their receptors involved with hematopoietic support and immunomodulatory properties of MSCs. Finally, the differentially expressed genes were collated for functional pathway enrichment analysis.\ud
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Results\ud
T1D-MSCs and C-MSCs were similar for morphology, immunophenotype, and differentiation potential. Our absolute gene expression results supported previous literature reports, while also detecting new potential molecules related to bone marrow-derived MSC functions. T1D-MSCs showed intrinsic abnormalities in mRNA expression, including the immunomodulatory molecules VCAM-1, CXCL12, HGF, and CCL2. Pathway analyses revealed activation of sympathetic nervous system and JAK STAT signaling in T1D-MSCs.\ud
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Conclusions\ud
Collectively, our results indicate that MSCs isolated from T1D patients present intrinsic transcriptional alterations that may affect their therapeutic potential. However, the implications of these abnormalities in T1D development as well as in the therapeutic efficacy of autologous MSCs require further investigation.This work was supported by research grants from by the following Brazilian\ud
institutions: National Council for Scientific and Technological Development\ud
(CNPq), São Paulo Research Foundation (FAPESP), and Agency for the\ud
Support and Evaluation of Graduate Education (CAPES)
Effectiveness of conventional treatment using bulk-fill composite resin versus Atraumatic Restorative Treatments in primary and permanent dentition: a pragmatic randomized clinical trial
No full textAbstract\ud
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Background\ud
Clinical studies are being conducted in less strict conditions in order to establish an adequate scientific basis for decision making. The aim of this pragmatic randomized clinical trial is to evaluate the effectiveness of single and multiple-surfaces restorations performed following the Atraumatic Restorative Treatment (ART) premises compared with Conventional Treatment (CT) using bulk fill composite restorations in primary and permanent teeth.\ud
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Methods/design\ud
A total of 1,214 5-to-13 year-old children with at least one single or multiple-surface dentin caries lesion in primary or permanent molars will be selected in public schools of Barueri-SP, Brazil. The participants will be randomly assigned into 2 groups: CT (caries removal with bur and restoration performed with Scotchbond™ Universal Adhesive system associated with Filtek Bulk Fill – 3 M/ESPE) and ART (Caries removal with hand instruments and restoration with high viscosity glass ionomer cement Ketac Molar Easy Mix – 3 M/ESPE). Ten untrained dentists will perform the treatment in in dental offices located at public schools. The restorations will be evaluated after 6, 12 and 24 months by an independent trained and calibrated examiner. The restoration and tooth survival, the cost-effectiveness analysis between the two groups and the operators’ preferences regarding the techniques will be also evaluated. Kaplan-Meier survival analysis and log-rank test will be applied for the restoration and tooth survival. All the average event rates in the two groups will be modelled and compared with a Cox proportional hazard shared frailty model since there is an operator-cluster effect. The significance level for all analyses will be 5 %.\ud
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Discussion\ud
Our hypothesis is that despite similar expected effectiveness between ART using high viscosity GIC and conventional treatment using bulk fill composite resin when treating single or multiple-surface in posterior primary and permanent teeth, ART will present superior cost-effectiveness. The results of this trial will support decision-making by clinicians and policy makers.\ud
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Trial registration\ud
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NCT02568917\ud
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. Registered on May 10th 2015.The authors would like to thank the Fundação de Amparo à Pesquisa do\ud
Estado de São Paulo – FAPESP for funding this paper and 3 M-ESPE for\ud
material donation
Respiratory allergy to Blomia tropicalis: Immune response in four syngeneic mouse strains and assessment of a low allergen-dose, short-term experimental model
No full textAbstract
Background
The dust mite Blomia tropicalis is an important source of aeroallergens in tropical areas. Although a mouse model for B. tropicalis extract (BtE)-induced asthma has been described, no study comparing different mouse strains in this asthma model has been reported. The relevance and reproducibility of experimental animal models of allergy depends on the genetic background of the animal, the molecular composition of the allergen and the experimental protocol.
Objectives
This work had two objectives. The first was to study the anti-B. tropicalis allergic responses in different mouse strains using a short-term model of respiratory allergy to BtE. This study included the comparison of the allergic responses elicited by BtE with those elicited by ovalbumin in mice of the strain that responded better to BtE sensitization. The second objective was to investigate whether the best responder mouse strain could be used in an experimental model of allergy employing relatively low BtE doses.
Methods
Groups of mice of four different syngeneic strains were sensitized subcutaneously with 100 μg of BtE on days 0 and 7 and challenged four times intranasally, at days 8, 10, 12, and 14, with 10 μg of BtE. A/J mice, that were the best responders to BtE sensitization, were used to compare the B. tropicalis-specific asthma experimental model with the conventional experimental model of ovalbumin (OVA)-specific asthma. A/J mice were also sensitized with a lower dose of BtE.
Results
Mice of all strains had lung inflammatory-cell infiltration and increased levels of anti-BtE IgE antibodies, but these responses were significantly more intense in A/J mice than in CBA/J, BALB/c or C57BL/6J mice. Immunization of A/J mice with BtE induced a more intense airway eosinophil influx, higher levels of total IgE, similar airway hyperreactivity to methacholine but less intense mucous production, and lower levels of specific IgE, IgG1 and IgG2 antibodies than sensitization with OVA. Finally, immunization with a relatively low BtE dose (10 μg per subcutaneous injection per mouse) was able to sensitize A/J mice, which were the best responders to high-dose BtE immunization, for the development of allergy-associated immune and lung inflammatory responses.
Conclusions
The described short-term model of BtE-induced allergic lung disease is reproducible in different syngeneic mouse strains, and mice of the A/J strain was the most responsive to it. In addition, it was shown that OVA and BtE induce quantitatively different immune responses in A/J mice and that the experimental model can be set up with low amounts of BtE
Impact of weight loss diet associated with flaxseed on inflammatory markers in men with cardiovascular risk factors: a clinical study
No full textAbstract
Background
Flaxseed has received attention for its anti-inflammatory and antioxidant role. The present study hypothesizes if flaxseed added to a weight loss diet could improve the lipid and metabolic profiles and decrease risk factors related to cardiovascular disease.
Methods
In a prospective, single blinded 42 days protocol, subjects were allocated into two groups with low carbohydrates intake: GriceLC (35% of carbohydrate and 60g of raw rice powder per day) and GflaxLC (32% of carbohydrate and 60g of flaxseed powder per day). Blood pressure, anthropometric measures and serum levels of isoprostane, C-reactive protein, Tumor Necrosis Factor-alpha, glucose, lipidic profile, uric acid, adiponectin, leptin and insulin were measured at baseline and at the end of interventions. Serum and urinary enterodiol and enterolactione were also measured.
Results
A total of 27 men with cardiovascular risk factors were evaluated, with mean age of 33 ± 10 years to GriceLC and 40 ± 9 years to GflaxLC. Both groups experienced weight loss and systolic blood pressure reduction. A decrease in inflammatory markers (CRP and TNF-α) was observed after flaxseed intake (mean decrease of 25% and 46% for GflaxLC respectively). All groups also showed improvement in levels of total cholesterol, LDL-c, uric acid and adiponectin. Only GflaxLC group showed a decrease in triglyceride levels.
Conclusion
This study suggests that flaxseed added to a weight loss diet could be an important nutritional strategy to reduce inflammation markers such as CRP and TNF-α.
Trial registration
ClinicalTrials.gov
NCT02132728
Speeding up all-pairwise dynamic time warping matrix calculation
No full textDynamic Time Warping (DTW) is certainly the most relevant distance for time series analysis. However, its quadratic time complexity may hamper its use, mainly in the analysis of large time series data. All the recent advances in speeding up the exact DTW calculation are confined to similarity search. However, there is a significant number of important algorithms including clustering and classification that require the pairwise distance matrix for all time series objects. The only techniques available to deal with this issue are constraint bands and DTW approximations. In this paper, we propose the first exact approach for speeding up the all-pairwise DTW matrix calculation. Our method is exact and may be applied in conjunction with constraint bands. We demonstrate that our algorithm reduces the runtime in approximately 50% on average and up to one order of magnitude in some datasets.FAPESP (grants #2013/26151-5 and #2015/07628-0)CNPq (grants #303083/2013-1 and #446330/2014-0
’HALITE IND. DS’: fast and scalable subspace clustering for multidimensional data streams
No full textGiven a data stream with many attributes and high frequency of events, how to cluster similar events? Can it be done in real time? For example, how to cluster decades of frequent measurements of tens of climatic attributes to aid real time alert systems in forecasting extreme climatic events, such as \ud
oods and hurricanes? The task of clustering data with many attributes is known as subspace clustering. Today, there exists a need for algorithms of this type well-suited to process multidimensional data streams, for which real time processing is highly desirable. This paper proposes the new algorithm 'HALITE ind. ds' - a fast, scalable and highly accurate subspace clustering algorithm for multidimensional data streams. It improves upon an existing technique that was originally designed to process static (not streams) data. Our main contributions are: (1) Analysis of Data Streams: the new algorithm takes advantage of the knowledge obtained from clustering past data to easy clustering data in the present. This fact allows our 'HALITE IND. DS' to be considerably faster than its base algorithm, yet obtaining the same accuracy of results; (2) Real Time Processing: as opposed to the state-of-the-art, 'HALITE IND. DS' is fast and scalable, making it feasible to analyze streams with many attributes and high frequency of events in real time; (3) Experiments: we ran experiments using synthetic data and a real multidimensional stream with almost one century of climatic data. Our 'HALITE IND. DS' was up to 217 times faster than 5 representative works, i.e., its base algorithm plus 4 others from the state-of-the-art, always presenting highly accurate results.FAPESPCAPESCNP