Immunohistochemical, tomographic and histological study on onlay bone graft remodeling. Part II: Calvarial bone

Objectives: Little information is available on the molecular events that occur during graft incorporation over time. The calvarial bone (Cb) grafts have been reported to produce greater responses compared with other donor regions in maxillofacial reconstructions, but the scientific evidences for this are still lacking. The objectives of this study are (1) to study the morphological pattern of Cb onlay bone grafts and compare them with the biological events through immunohistochemical responses and (2) to establish the effects of perforations in maintaining the volume and bone density of the receptor bed. Material and methods: Sixty New Zealand White rabbits were submitted to Cb onlay bone grafts on the mandible. In 30 rabbits, the receptor bed was perforated (perforated group), while for the remaining animals the bed was kept intact (non-perforated group). Six animals from each group were sacrificed at 5, 7, 10, 20 and 60 days after surgery. Histological sections from the grafted area were prepared for immunohistochemical and histological analyses. Immuno-labeling was found for proteins Osteoprotegerin (OPG), receptor activator of nuclear factor-κβ ligand (RANKL), alkaline phosphatase (ALP), osteopontin (OPN), vascular endothelial growth factor (VEGF), tartrate-resistant acid phosphatase (TRAP), Type I collagen (COL I) and osteocalcin (OC). The tomography examination [computerized tomography (CT) scan] was conducted just after surgery and at the sacrifice. Results: The histological findings revealed that the perforations contributed to higher bone deposition during the initial stages at the graft-receptor bed interface, accelerating the graft incorporation process. The results of the CT scan showed lower resorption for the perforated group (P≤0.05), and both groups showed high bone density rates at 60 days. This set of evidences is corroborated by the immunohistochemical outcomes indicating that proteins associated with revascularization and osteogenesis (VEGF, OPN, TRAP and ALP) were found in higher levels in the perforated group. Conclusions: These findings indicate that the bone volume of calvarial grafts is better maintained when the receptor bed is perforated, probably resulting from more effective graft revascularization and greater bone deposition. The process of bone resorption peaked between 20 and 60 days post-operatively in both groups although significantly less in the perforated group. © 2009 John Wiley & Sons A/S.

Influence of environmental temperature, dietary energy level and sex on performance and carcass characteristics of pigs

Pigs are quite sensitive to high environmental temperatures and the thermoregulation mechanisms represent great expenses in energy for heating loss, reducing animal well-being and production performance, and altering carcass quality. The aim of this study was to assess the effects of sex and dietary energy level in growing-finishing pigs submitted to characteristic seasonal variation of temperature in subtropical humid climate, and to propose a mathematical model to predict growth performance and carcass characteristics. Twenty-eight crossbred growing-finishing pigs were randomly allotted to twelve treatments, in a 2x2x3 factorial trial (2 sex; 2 environmental conditions, and 3 energy levels). Heat stress condition (climatic chamber) showed temperatures of 31 oC at 7:00 and 22 oC at 17:00 (maximum of 33 °C) and thermal comfort condition (stall) showed temperatures of 18 °C at 7:00 and 24 °C (maximum of 27 °C). Pigs were fed ad libitum with diets containing 12.2 (low), 13.6 (medium) and 15.0 (high) MJ ME/ kg DM. Voluntary feed intake, daily weight gain, and final body weight were higher (P<0.01) at thermal comfort condition and were influenced by sex (P<0.01) in growing pigs. Feed to gain ratio decreased as the energy level increased (P<0.01), with values of 2.67, 2.59, and 2.32 (12.2, 13.6, and 15.0 MJ ME/kg DM, respectively). There was energy level and sex interaction only for daily weight gain. Regarding finishing pigs, environmental conditions also showed effects (P<0.01) on voluntary feed intake, daily weight gain, and final body weight. Performance of pigs was better at thermal comfort condition. Feed to gain ratio values were 3.55, 3.42, and 2.95 for low, medium, and high energy level, respectively. Interactions between energy level and sex were observed for voluntary feed intake, daily weight gain, and final body weight (P<0.05). Carcass yield and quality were affected by environmental condition and dietary energy level. Both hot and cold carcass weight increased as energy of ration increased. Cold carcass weight increased by 1.142 kg/MJ EM whereas backfat thickness was up to 252 mm/MJ EM. Longissimus thoracis muscle thickness was around 16 mm smaller in pigs under heat stress, but lean content was 2.68% higher in those animals. Regression equations were proposed to predict the performance values in the different situations studied.

Utilização de extratos de plantas medicinais e óleo de Eucaliptus no controle in vitro de Microsporum canis

INTRODUCTION: Microsporum canis is the most common cause of canine and feline dermatophytosis and thus has an important zoonotic role. OBJECTIVES: the aim of this study was to determine the antifungal action of medicinal plant extracts and of eucalyptus oil against pathogenic fungus Microsporum canis. METHODS: the extracts were prepared by mixing 300 g of previously washed leaves with 450 mL of distilled water. Then the material was triturated, filtered, sterilized and conserved at 10 + 2 oC. Fifteen milliliters of sterilized medium Sabouraud dextrose (Difco) at a temperature of 55 + 1 oC was added in Petri dishes containing the extracts in one, two, three, four and five mm concentrations. The fungus was inoculated once the medium was solidified. The inoculated dishes were maintained in B.O.D. incubator at 36 ± 0,5 oC until the fungus developed in the controls. RESULTS: the extracts from Punica granatum, Mangifera indica and Eucalyptus spp reduced the growth of fungus, but the extracts from Cymgopogom nardus, Tagetes minuta, Ruta graviolens, Cyperus rotundus, Annona moricata and Calendula spp leaves and flowers boosted the growth of fungus. The other extracts and the eucalyptus oil neither show any fungicidal action nor encourage mycelium growth. CONCLUSIONS: the use of most tested extracts and eucalyptus oil is not suitable for the treatment of Microsporum canis dermatophytosis due to lack of inhibitory effects. The extracts from Cymgopogom nardus, Tagetes minuta, Ruta graviolens, Cyperus rotundus, Annona moricata and from of Calendula spp leaves and flowers help the development of the fungus making clear that phytotherapy should be properly used, otherwise it can worsen the problem. However; extracts from Mangifera indica, Punica granatum and Eucalyptus spp. can be used as fungistatic.

Viabilidade de sementes de Piptadenia moniliformis Benth. Pelo teste de tetrazólio

Piptadenia moniliformis Benth. is a tree species which is important for beekeeping, as well as being recommended for soil restoration, reforestation, wood production for small civil construction projects, and cattle and sheep forage. Information on how to evaluate seed physiological quality is still scarce and in this was the study aimed to adapt the procedures of the tetrazolium test to assess the viability of P. moniliformis seeds. Four seed lots were scarified in sulphuric acid for 30 min, and soaked between paper towels at 25 °C for 24 hours. The seed coat was then removed and the naked seeds immersed in tetrazolium solutions with concentrations of 0.05, 0.075 and 0.1% for 2, 3, and 4 hours at 35 oC in the dark. Each treatment consisted of four replications of 25 seeds. The embryos were classified according to viability based on the staining patterns. The previous soaking of the seeds for 24 hours at 25 oC between paper towels, followed by the removal of the seed coat and staining of the naked seeds for 4 hours in a 0.075% tetrazolium solution at 35 oC was the most efficient method for evaluating the viability of P. moniliformis Benth seeds.

Influência de diferentes sistemas adesivos na resistência de união de pinos de fibra à dentina intrarradicular

Objective: To evaluate the influence of three adhesive systems on the bond strength of fiber post luted to root dentine. The hypothesis was that the bond strength is influenced by the adhesive system. Method: The canals of thirty single-root bovine roots (16mm in length) were prepared using the preparation drill (FGM) until 12mm. 14 mm of each root was embedded with acrylic resin and the specimens were allocated into three groups (n=10), considering the factor adhesive (3 levels): Gr1- Scotchbond Multipurpose Plus (3M ESPE), Gr2- One Step (Bisco) and Gr3- Excite DSC (Ivoclar Vivadent). The adhesive systems were applied using a microbrush, according to the manufacture's recommendations. The fiber posts (White Post DC, FGM) were luted with dual resin cement (All-Cem,FGM). After, the cores with composite resin (Llis, FGM) were made and each set of root/post/core was submitted to the mechanical cycling (Erios, Brazil) (10 6 cycles, 84N, 4 Hz, inclination of 45 o, 37 oC, water). Each specimen was cut in 4 samples (1.8mm in thickness), which were submitted to the push-out test in a universal testing machine (ServoPulser - Shimadzu) (50Kgf, 1mm/min). The data (MPa) were analyzed using ANOVA (1-way) and Tukey test (5%). Results: The factor adhesive (P=0.00352) influenced the bond strength significantly (ANOVA). Gr1 (6.8±3.8 MPa) a presented higher bond strength values than Gr2 (3.1±1.5 MPa) b and similar to Gr3 (4.4±3.3) a,b. Moreover, Gr3 and Gr2 were similar between them (Tukey). The hypothesis was accepted. Conclusion: Based on the results it was concluded that chemical and dual polymerization adhesive system should be used for the adhesive luting fiber post procedures.

Intussuscepção ileocólica associada a torção mesentérica em um cão: Relato de caso

Mesenteric torsion is a rare disease in dogs whereas it is more common to affect large animals. The main clinical signs observed are acute, such as prostration and abdominal distension which evolve to a state of shock and death. This paper reports the mesenteric torsion in a dog that was presented to veterinary care with generalized acute abdominal pain of sudden onset. At the physical examination, signs of acute abdomen, rectal temperature of 35 oC, moderate dehydration, congested mucous membrane, and prostration were detected. Emergency therapy was instituted and radiographic and ultrasound examinations were requested. The radiography showed increase in the bowel diameter and an image suggestive of intussusception. The patient was referred to emergency exploratory laparotomy and obtained a definite diagnosis of ileocolic intussusception associated to mesenteric torsion. Due to the extent of intestinal dysfunction, the patient was euthanized on the operating table.

Assessment of force decay in orthodontic elastomeric chains: An in vitro study

Introduction: Elastomeric materials are considered important sources of orthodontic forces. Objective: To assess force degradation over time of four commercially available orthodontic elastomeric chains (Morelli, Ormco, TP and Unitek). Methods: The synthetic elastics were submerged in 37 oC synthetic saliva and stretched by a force of 150 g (15 mm - Morelli and TP; 16mm - Unitek and Ormco). With a dynamometer, the delivered force was evaluated at different intervals: 30 minutes, 7 days, 14 days and 21 days. The results were subjected to ANOVA and Tukey's test. Results: There was a force decay between 19% to 26.67% after 30 minutes, and 36.67% to 57% after 21 days of activation. Conclusions: TP elastomeric chains exhibited the smallest percentage of force decay, with greater stability at all time intervals tested. Meanwhile, the Unitek chains displayed the highest percentage of force degradation, and no statically significant difference was found in force decay between Ormco and Morelli elastomeric chains during the study period.

Histomorphometrical and molecular evaluation of endosseous dental implants sites in humans: Correlation with clinical and radiographic aspects

Objective: To evaluate the correlations between clinical-radiographical aspects and histomorphometric-molecular parameters of endosseous dental implant sites in humans. Material and methods: The study sample consisted of bone implant sites from the jawbones of 32 volunteers, which were classified according to two different systems: (1) based only on periapical and panoramic images (PP); (2) as proposed by Lekholm & Zarb (L&Z). Bone biopsies were removed using trephine during the first drilling for implant placement. Samples were stained with haematoxylin-eosin (HE), and histomorphometric analysis was performed to obtain the following parameters: trabecular thickness (Tb.Th), trabecular number, bone volume density (BV/TV), bone specific surface (BS/BV), bone surface density and trabecular separation (Tb.Sp). In addition, immunohistochemistry analysis was performed on bone tissue samples for the proteins, Receptor activator of nuclear factor kappa-B (RANK), RANK ligand (RANKL), osteoprotegerin (OPG) and Osteocalcin (OC). Also, the determination of the relative levels of gene expression was performed using Reverse transcription-real-time Polymerase Chain Reaction (RT-PCR). Results: PP and L&Z classification systems revealed a moderate correlation with BV/TV, BS/BV, Tb.Th and Tb.Sp. L&Z's system identified differences among bone types when BV/TV, BS/BV, Tb.Th and Tb.Sp were compared. A weak correlation between PP/L&Z classifications and the expression of bone metabolism regulators (RANK, RANKL, OPG e OC) was found. The analysis of mRNA expression showed no difference between the bone types evaluated. Conclusions: Our results suggest that PP and L&Z subjective bone-type classification systems are related to histomorphometric aspects. These data may contribute to the validation of these classifications. Bone remodelling regulatory molecules do not seem to influence morphological aspects of the jawbone © 2011 John Wiley & Sons A/S.

PLA and Montmorilonite Nanocomposites: Properties, Biodegradation and Potential Toxicity

The concern related to solid waste increases efforts to develop products based on biodegradable materials. At present, PLA has one of the highest potentials among biopolyesters, particularly for packaging. However, its application is limited in some fields. In order to optimize PLA properties, organo-modified montmorilonites have been extensively used to obtain nanocomposites. Although PLA nanocomposites studies are widely reported in the literature, there is still few information about the influence of organoclays on de biodegradation process, which is a relevant information, since one of the main purposals related to the final disposal of biopolymers as PLA is composting. Besides, in the last years some research has been conducted in order to evaluate the potential toxicity of montmorilonite, unmodified or organo-modified. Since the use of montmorilonite is expanding in different applications, human exposure and risk assessment are important issues to be investigated. In this context, this review intends to compile available information related to common organoclays used for PLA nanocomposites, its properties, biodegradation analysis and potential toxicity evaluation of nanocomposites, focused on montmorilonite as filler. Two issues of relevance were pointed out. The first is food safety and quality, and the second consideration is the environmental effect. © 2013 Springer Science+Business Media New York.

Sorption and desorption of phosphate on biochar and biochar-soil mixtures

The term biochar refers to materials with diverse chemical, physical and physicochemical characteristics that have potential as a soil amendment. The purpose of this study was to investigate the P sorption/desorption properties of various slow biochars and one fast pyrolysis biochar and to determine how a fast pyrolysis biochar influences these properties in a degraded tropical soil. The fast pyrolysis biochar was a mixture of three separate biochars: sawdust, elephant grass and sugar cane leaves. Three other biochars were made by slow pyrolysis from three Amazonian tree species (Lacre, Ingá and Embaúba) at three temperatures of formation (400 °C, 500 °C, 600 °C). Inorganic P was added to develop sorption curves and then desorbed to develop desorption curves for all biochar situations. For the slow pyrolysis, the 600 oC biochar had a reduced capacity to sorb P (4-10 times less) relative to those biochars formed at 400 °C and 500 °C. Conversely, biochar from Ingá desorbed the most P. The fast pyrolysis biochar, when mixed with degraded tropical mineral soil, decreased the soil's P sorption capacity by 55% presumably because of the high soluble, inorganic P prevalent in this biochar (909 mg P/kg of biochar). Phosphorus desorption from the fast pyrolysis biochar/soil mixture not only exhibited a common desorption curve but also buffered the soil solution at a value of ca. 0.2 mg/L. This study shows the diversity in P chemistry that can be expected when biochar is a soil amendment and suggests the potential to develop biochars with properties to meet specific objectives. © 2013 British Society of Soil Science.

UV filters in water samples: Experimental design on the SPE optimization followed by GC-MS/MS analysis

Esse estudo descreve o desenvolvimento e otimização de um método de extração em fase solida (SPE) para análise dos filtros ultravioletas (UV): benzofenona-3 (BP-3), etilhexil salicilato (ES), etilhexil metoxinamato (EHMC) e octocrileno (OC) em matrizes ambientais. Um planejamento fatorial fracionário (PFF) 25-1 foi empregado na avaliação das variáveis significativas do método de extração. As condições experimentais otimizadas da avaliação estatística foram: capacidade do cartucho de 500 mL, eluente acetato de etila, metanol como solvente de lavagem (10% em água, v/v) and volume do eluente de 3 × 2 mL e pH 3. Os parâmetros analíticos avaliados foram satisfatõrios, apresentando linearidade de 100 a 4000 ng L -1, recuperaç ões para os quatro níveis de fortificação (Limite de Quantificação do Método, 200, 1000 e 2000 ng L-1) entre 62 e 107% com desvio padrão relativo menor que 14%. Os limites de quantificação foram encontrados na faixa de ng L-1, variando entre 10 e 100 ng L-1. O método proposto foi aplicado para a determinação dos quatro filtros UV em amostras de águas naturais. This study describes the development and optimization of a solid-phase extraction (SPE) method for analysis of ultraviolet (UV) filters, benzophenone-3 (BP-3), ethylhexyl methoxycinnamate (EHMC), ethylhexyl salicylate (ES) and octocrylene (OC), in environmental matrices. A 25-1 fractional factorial design (FFD) was used to evaluate the significant variables for the extraction method. The optimized experimental conditions determined from the statistical evaluation were: breakthrough volume of 500 mL, eluent of ethyl acetate, wash solvent of methanol (10% in water, v/v), eluent volume of 3 × 2 mL and pH 3. The evaluated analytical parameters were satisfactory for the analytes and showed linearity between 100 and 4000 ng L-1, recoveries for four fortification levels (Method Quantification Limit, 200, 1000 and 2000 ng L-1) were between 62 and 107% with relative standard deviations less than 14%. Limits of quantification were in the ng L-1 range and were between 10 and 100 ng L-1. The proposed method was used to analyze four UV filters in natural water samples. ©2013 Sociedade Brasileira de Química.

Dynamics of bone healing after osteotomy with piezosurgery or conventional drilling - histomorphometrical, immunohistochemical, and molecular analysis

Background: Piezosurgery is an osteotomy system used in medical and dental surgery. Many studies have proven clinical advantages of piezosurgery in terms of quality of cut, maneuverability, ease of use, and safety. However, few investigations have tested its superiority over the traditional osteotomy systems in terms of dynamics of bone healing. Therefore, the aim of this study was to evaluate the dynamics of bone healing after osteotomies with piezosurgery and to compare them with those associated to traditional bone drilling.Methods: One hundred and ten rats were divided into two groups with 55 animals each. The animals were anesthetized and the tibiae were surgically exposed to create defects 2 mm in diameter by using piezosurgery (Piezo group) and conventional drilling (Drill group). Animals were sacrificed at 3, 7, 14, 30 and 60 days post-surgery. Bone samples were collected and processed for histological, histomorphometrical, immunohistochemical, and molecular analysis. The histological analysis was performed at all time points (n = 8) whereas the histomorphometrical analysis was performed at 7, 14, 30 and 60 days post-surgery (n = 8). The immunolabeling was performed to detect Vascular Endothelial Growth Factor (VEGF), Caspase-3 (CAS-3), Osteoprotegerin (OPG), Receptor Activator of Nuclear Factor kappa-B Ligand (RANKL), and Osteocalcin (OC) at 3, 7, and 14 days (n = 3). For the molecular analysis, animals were sacrificed at 3, 7 and 14 days, total RNA was collected, and quantification of the expression of 21 genes related to BMP signaling, Wnt signaling, inflammation, osteogenenic and apoptotic pathways was performed by qRT-PCR (n = 5).Results: Histologically and histomorphometrically, bone healing was similar in both groups with the exception of a slightly higher amount of newly formed bone observed at 30 days after piezosurgery (p < 0.05). Immunohistochemical and qRT-PCR analyses didn't detect significant differences in expression of all the proteins and most of the genes tested.Conclusions: Based on the results of our study we conclude that in a rat tibial bone defect model the bone healing dynamics after piezosurgery are comparable to those observed with conventional drilling. © 2013 Esteves et al.; licensee BioMed Central Ltd.

Effect of forced-molting methods and rearing temperatures on the performance and organ biometrics of laying hens

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comportamento hemodinâmico e metabólico do choque hemorrágico: estudo experimental no cão

Pós-graduação em Anestesiologia - FMB

Avaliação do desempenho de tilápias (Oreochromis niloticus, Trewavas, 1983) em tanques-rede na represa de Jurumirim/Alto Rio Paranapanema

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)