110 resultados para Preservation of embryo
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The structure and ultrastructure of immature to fully mature glandular dots in the leaf, floral organs and fruit, and their secretion components were described in Caesalpinia echinata Lam. (Leguminosae) for the first time. Data showed that glandular dots were groups of idioblasts with contents that reacted positively for both lipophilic and hydrophilic substances. Idioblasts originated from successive divisions of the ground meristem cells or mesophyll cells of an ovary of a fertilized flower. Following division, cells enlarged, the cytoplasm became denser and its content became full. No idioblasts were observed after fruit sclerification. Besides these mixed-content idioblasts, some cells in the sepals, petals and mesocarp were found to contain phenolic compounds, which probably represent a kind of constitutive defense mechanism, once the flowers and fruits become highly fitness-valued parts of the plant and can be commonly attacked. The contents of the idioblasts are released as the growth rate of the embryo increases, indicating that the plant probably diverts the precursors of secondary metabolites into the primary metabolism, at this critical time of embryo development.
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The aim of this article is to present the decalcification process dynamic of mineralized tissue in dogs, teeth and jaw, comparing the traditional decalcification method, immersion, and microwave, immersion followed by irradiation using a domestic microwave oven, accompanying the liberation of calcium through spectrophotometer of atomic absorption. It was used as decalcified agent, EDTA solution or nitric acid. The results showed that with the use of nitric acid (5%), after 15 days, the irradiated fragments could be processed for histological analysis, otherwise the tooth not irradiated need to be submerged for 65 days. The EDTA decalcified action was slower than the nitric acid. The histological observations of the irradiated samples showed an excellent preservation of the morphological characteristics, independently of the decalcified agent used. © 2007 Sociedad Chilena de Anatomía.
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Bone decalcification is a time-consuming process. It takes weeks and preservation of the tissue structure depends on the quality and velocity of the demineralization process. In the present study, a decalcification methodology was adapted using microwaving to accelerate the decalcification of rat bone for electron microscopic analysis. The ultrastructure of the bone decalcified by microwave energy was observed. Wistar rats were perfused with paraformaldehyde and maxillary segments were removed and fixed in glutaraldehyde. Half of specimens were decalcified by conventional treatment with immersion in Warshawsky solution at 4oC during 45 days, and the other half of specimens were placed into the beaker with 20 mL of the Warshawsky solution in ice bath and thereafter submitted to irradiation in a domestic microwave oven (700 maximum power) during 20 s/350 W/±37°C. In the first day, the specimens were irradiated 9 times and stored at 40°C overnight. In the second day, the specimens were irradiated 20 times changing the solution and the ice after each bath. After decalcification, some specimens were postfixed in osmium tetroxide and others in osmium tetroxide and potassium pyroantimonate. The specimens were observed under transmission electron microscopy. The results showed an increase in the decalcification rate in the specimens activated by microwaving and a reduction of total experiment time from 45 days in the conventional method to 48 hours in the microwave-aided method.
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Although there are almost thirty-thousand species of fish living in a great variety of habitats and utilizing vast reproductive strategies, our knowledge of morphofunctional and quantitative aspects of testis structure and spermatogenesis is still incipient for this group of vertebrates. In this review, we discuss aspects that are important to better understanding of testis structure and function, and of the development of germ cells (GC) during spermatogenesis. To achieve this, we have recently completed a number of studies presenting morphometric and functional data related to the numbers of GC and Sertoli cells (SC) per each type of spermatogenic cyst, the number of spermatogonial generations, the SC efficiency, and the magnitude of GC loss that normally occurs during spermatogenesis. We also investigated SC proliferation and the relationship of this important event to early spermatogenic cysts. The available data strongly suggest that SC proliferation in sexually mature tilapia is the primary factor responsible for the increase in testis size and for determination of the magnitude of sperm production. The influence of temperature on the duration of spermatogenesis in tilapia was also evaluated and we have used this knowledge to deplete endogenous spermatogenesis in this teleost, in order to develop an experimental system for GC transplantation. This exciting technique results in new possibilities for investigation of spermatogenesis and spermatogonial stem cell biology, creating also an entirely new and promising scenario in biotechnology - transgenic animal production and the preservation of the genetic stocks of valuable animals or endangered species. © Springer Science+Business Media B.V. 2008.
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According to the environmental legislation enforced in Brazil and the process of marketing globalization, the commitment of the nations to the preservation of the environment is intensified. By reason of nature's negative responses to its intensive use, awareness then appears from enterprises and agencies about how the anthropic action over the environment needs to be minimized, becoming a challenge: development and sustainability. In this context, the present work made use of the Mechanical tillage of the soil, as a technique to apply, in a large scale, the strategies and methods to recover mined areas that were researched and developed experimentally by researchers on a theme project about the recovering of degraded areas. This work was conducted in the Amazon ecosystem, inside the Jamari National Forest - Rondônia (FLONA do Jamari), in deactivated cassiterite mines. The objectives of this work were to: Develop a computational program capable of managing a database and assist in the selection of machines and preparation methods to execute the operations of topographical reconstitution and tillage of surfaces in areas degraded by the mineral exploitation of cassiterite. Use the program that was developed in the planning of costs and operational development, for the operations required in the strategies for recovering the areas. Analyze the vegetable productivity in the mobilized areas and the quality of the superficial mobilization, making use of indicators and tillage methods. Evaluate, through biological indicators, the efficiency of the recovery strategies and techniques that were mechanized and applied on the location. The results showed that the developed computational program (SGMAD) served the methodological purposes (the analysis of costs and operational capacity) established for the planning and the selection of the tillage machines and methods in the areas of mineral exploitation of cassiterite. The applied methods and quality of the superficial mobilization were significant to the development of leguminous plants in the areas. The use of biological indicators (microbial biomass and enzymatic activity) in the evaluation of the adopted techniques and strategies revealed that the planting of leguminous plants and their posterior incorporation have been promoting gradually positive alterations in some of the analyzed soil/substract parameters. © 2010 WIT Press.
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Two discussions are imposed to the designers of wood constructions. The first one deals with the technical knowledge to project and execute buildings; the second one is concerned with the preservation of the environment, use of the wood in a sustainable way. The work presents the tendencies of the wood used in the Brazilian habitation architecture characterizing the used woods; how the construction technical solutions have developed and discusses about the necessity of using the wood in a conscious way, knowing its origin and control, sustainable use of the forests resources. It focalizes, mainly, the search of the designers to harmonize the use of the wood and the preservation of the forest biodiversity, when it deals with the native forest.
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Objective: Although direct bonding takes up less clinical time and ensures increased preservation of gingival health, the banding of molar teeth is still widespread nowadays. It would therefore be convenient to devise methods capable of increasing the efficiency of this procedure, notably for teeth subjected to substantial masticatory impact, such as molars. This study was conducted with the purpose of evaluating whether direct bonding would benefit from the application of an additional layer of resin to the occlusal surfaces of the tube/tooth interface. Methods: A sample of 40 mandibular third molars was selected and randomly divided into two groups: Group 1 - Conventional direct bonding, followed by the application of a layer of resin to the occlusal surfaces of the tube/tooth interface, and Group 2 - Conventional direct bonding. Shear bond strength was tested 24 hours after bonding with the aid of a universal testing machine operating at a speed of 0.5mm/min. The results were analyzed using the independent t-test. Results: The shear bond strength tests yielded the following mean values: 17.08 MPa for Group 1 and 12.60 MPa for Group 2. Group 1 showed higher statistically significant shear bond strength than Group 2. Conclusions: The application of an additional layer of resin to the occlusal surfaces of the tube/tooth interface was found to enhance bond strength quality of orthodontic buccal tubes bonded directly to molar teeth.
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Background: Throughout dairy cows evolution, milk production was always the key point to select the superior animal. Currently, several evidences has shown that high milk production have intensively contributed to the decline of dairy cattle fertility. Beyond milk production, dairy cows have their reproductive performance impaired by another factors, heat stress and repeat-breeding. Methods like fixed time artificial insemination and embryo transfer were developed to minimize the effects of these factors, and improve dairy herds profitability. This review aims to show some key-point experiments conducted to improve the efficiency of the self-appointed protocols for artificial insemination and embryo transfer in Brazil, overcoming several reproductive problems. Our goal is to develop cheap and easy self-appointed programs that facilitate animal handling and maximize their reproductive outcomes all over the year. Review: Failure in estrus detection is the mainly limiting factor for the use of artificial insemination in high-production dairy herd. An excellent alternative to overcome the need of estrus detection is fixed time artificial insemination. Many protocols with and without the use of estradiol have been developed to that end. Among the protocols for fixed time artificial insemination without estradiol, DoubleOvsynch has been extensively used recently in American dairy herds. In Brazil, similar pregnancy rate was obtained compared to progesterone-estradiol based protocols for fixed time artificial insemination. Particularities of progesterone-estradiol based protocols as (1) new progesterone device or devices previously used for eight days; (2) different doses of eCG; and (3) the use of estradiol cypionate for fixed time artificial insemination have been studied in Brazil. The use of self-appointed artificial insemination also enabled the reduction of the interval calving-conception compared to cows inseminated following the standing estrus. Regarding the low fertility of repeat breeders and the effect of heat stress at early pregnancy, other methods like embryo transfer became important tools to enhance reproductive efficiency of Brazilian dairy herds. Protocols were also developed to allow fixed time embryo transfer, eliminating the need of estrus detection and improving the reproductive efficiency of lactating recipients. As well as described for fixed time artificial insemination treatments, there is a large variety of hormone combination for fixed time embryo transfer (with and without estradiol). An experiment conducted in Brazil demonstrated that protocols for fixed time embryo transfer without estradiol can be as good as with estradiol to synchronize high-producing Holstein recipients, essentially during summer. Particularities related to embryos cryopreservation, synchronization of the estrus cycle of donors and recipients and the site of embryo release into the uterine horn were also investigated. Greater conception rates were achieved when fresh embryos were transferred compared to frozen-thawed ones. Also, the tight synchronization between donor and recipient (same day of estrus) resulted more pregnancies than when recipients were one day later or in advantage in relation to donors. Moreover, the site of embryo release into the uterine horn (ipsilateral to the corpus luteum) had no effect on pregnancy rates after in vivo produced embryo transfer. Conclusion: Both fixed time artificial insemination and fixed time embryo transfer are important tools to improve reproductive efficiency of high-producing dairy cows. These biotechnologies help bypassing some of the greatest challenges of dairy cattle reproduction: the difficulties of estrus detection, and the low fertility associated to heat stress and repeat breeding.
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Summary Trichostatin A (TSA) is a histone deacetylase inhibitor that induces histone hyperacetylation and increases gene expression levels. The aim of the present study was to establish a suitable condition for the use of TSA in in vitro cultures of bovine embryos, and to determine whether TSA would increase blastocyst rates by improvement of chromatin remodelling during embryonic genome activation and by increasing the expression of crucial genes during early development. To test this hypothesis, 8-cell embryos were exposed to four concentrations of TSA for different periods of time to establish adequate protocols. In a second experiment, three experimental groups were selected for the evaluation of embryo quality based on the following parameters: apoptosis, total cell number and blastocyst hatching. TSA promoted embryonic arrest and degeneration at concentrations of 15, 25 and 50 nM. All treated groups presented lower blastocyst rates. Exposure of embryos to 5 nM for 144 h and to 15 nM for 48 h decreased blastocyst hatching. However, the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay revealed similar apoptosis rates and total cell numbers in all groups studied. Although, in the present study, TSA treatment did not improve the parameters studied, the results provided background information on TSA supplementation during in vitro culture of bovine embryos and showed that embryo quality was apparently not affected, despite a decrease in blastocyst rate after exposure to TSA. © Cambridge University Press 2011.
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The recovery of sperm from the epididymal cauda may be the last chance to obtain genetic material when sudden death or serious injuries occur in valuable stallions. However, the lack of technical knowledge regarding the storage and transportation of the epididymis often prevents the preservation of the sperm. Therefore, the aim of this study was to compare sperm parameters of sperm obtained immediately after orchiectomy with sperm recovered from epididymal cauda at different times after storage at 5°C and at room temperature (RT). For that, 48 stallions of different breeds were used. In group 1 (control group), eight stallions were used, and the harvest of the epididymal sperm was performed immediately after orchiectomy. In group 2, 40 stallions were used, which were divided into five groups according to the storage time of the epididymis after orchiectomy (6, 12, 18, 24, or 30 hours), making a total of eight stallions per group. One epididymis of each stallion was stored at 5°C, and the contralateral epididymis was stored at RT, both for the same period. The sperm parameters of total motility, progressive motility, progressive linear velocity, curvilinear velocity, percentage of rapid sperm, and plasma membrane integrity were evaluated in all the groups after sperm recovery, resuspension in a sperm freezing diluent, and thawing. In conclusion, the storage of the testis-epididymis complex at 5°C provided better preservation of epididymal sperm than the storage at RT, and regardless of the temperature, the progressive motility is the sperm parameter that is most sensitive to storage time. © 2013 Elsevier Inc.
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Background:Ventral root avulsion is an experimental model of proximal axonal injury at the central/peripheral nervous system interface that results in paralysis and poor clinical outcome after restorative surgery. Root reimplantation may decrease neuronal degeneration in such cases. We describe the use of a snake venom-derived fibrin sealant during surgical reconnection of avulsed roots at the spinal cord surface. The present work investigates the effects of this fibrin sealant on functional recovery, neuronal survival, synaptic plasticity, and glial reaction in the spinal motoneuron microenvironment after ventral root reimplantation.Methodology/Principal Findings:Female Lewis rats (7 weeks old) were subjected to VRA and root replantation. The animals were divided into two groups: 1) avulsion only and 2) replanted roots with fibrin sealant derived from snake venom. Post-surgical motor performance was evaluated using the CatWalk system twice a week for 12 weeks. The rats were sacrificed 12 weeks after surgery, and their lumbar intumescences were processed for motoneuron counting and immunohistochemistry (GFAP, Iba-1 and synaptophysin antisera). Array based qRT-PCR was used to evaluate gene regulation of several neurotrophic factors and receptors as well as inflammatory related molecules. The results indicated that the root reimplantation with fibrin sealant enhanced motor recovery, preserved the synaptic covering of the motoneurons and improved neuronal survival. The replanted group did not show significant changes in microglial response compared to VRA-only. However, the astroglial reaction was significantly reduced in this group.Conclusions/Significance:In conclusion, the present data suggest that the repair of avulsed roots with snake venom fibrin glue at the exact point of detachment results in neuroprotection and preservation of the synaptic network at the microenvironment of the lesioned motoneurons. Also such procedure reduced the astroglial reaction and increased mRNA levels to neurotrophins and anti-inflammatory cytokines that may in turn, contribute to improving recovery of motor function. © 2013 Barbizan et al.
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Studies to determine mite species richness in natural environments are still scarce, and have been conducted mainly in tropical ecosystems. The aim of this study was to determine the species richness of mites on two common native plants in fragments of the semideciduous seasonal forest in the Northwest of São Paulo State, Brazil. In each of eight fragments, 10 specimens of Actinostemon communis (Euphorbiaceae) and 10 of Trichilia casaretti (Meliaceae) were selected and marked. In total, 124 species of mites belonging to 21 families were found on the two plants. Tarsonemidae had the highest diversity (34 species), followed by Phytoseiidae (31), Tetranychidae (9) and Tenuipalpidae (8). Species accumulation curves for the two sampled plants did not reach an asymptote, even with the large sampling effort. Hence, it is estimated that a greater sampling effort may lead to an increase in species richness compared with what was found in this study. The richness of this mite fauna suggests that preservation of these plant species is important to maintain the mite diversity in these forest fragments. © 2013 Taylor & Francis Group, LLC.
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
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Objective The objective was to examine the effect of a solvent dimethyl sulfoxide (DMSO) on resin-dentin bond durability, as well as potential functional mechanisms behind the effect. Methods Microtensile bond strength (μTBS) was evaluated in extracted human teeth in two separate experiments. Dentin specimens were acid-etched and assigned to pre-treatment with 0.5 mM (0.004%) DMSO as additional primer for 30 s and to controls with water pre-treatment. Two-step etch-and-rinse adhesive (Scotchbond 1XT, 3M ESPE) was applied and resin composite build-ups were created. Specimens were immediately tested for μTBS or stored in artificial saliva for 6 and 12 months prior to testing. Additional immediate and 6-month specimens were examined for interfacial nanoleakage analysis under SEM. Matrix metalloproteinase (MMP) inhibition by DMSO was examined with gelatin zymography. Demineralized dentin disks were incubated in 100% DMSO to observe the optical clearing effect. Results The use of 0.5 mM DMSO had no effect on immediate bond strength or nanoleakage. In controls, μTBS decreased significantly after storage, but increased significantly in DMSO-treated group. The control group had significantly lower μTBS than DMSO-group after 6 and 12 months. DMSO also eliminated the increase in nanoleakage seen in controls. 5% and higher DMSO concentrations significantly inhibited the gelatinases. DMSO induced optical clearing effect demonstrating collagen dissociation. Significance DMSO as a solvent may be useful in improving the preservation of long-term dentin-adhesive bond strength. The effect may relate to dentinal enzyme inhibition or improved wetting of collagen by adhesives. The collagen dissociation required much higher DMSO concentrations than the 0.5 mM DMSO used for bonding. © 2013 Academy of Dental Materials.