Search

Evaluate the effect of 12 wks of concurrent training (CT) in the extracellular matrix (ECM) of subcutaneous adipose tissue (SAT) in people living with HIV/AIDS (PLWHA). In the non-randomized clinical trial, 19 participants, 11 healthy (HIV–) and 18 PLWHA under the use of highly active antiretroviral therapy (HAART) for at least 1 year (HIV+). All participants engaged in a moderate-intensity CT program for 12 weeks, 3 times a week. Before and after CT, aerobic and strength performance were assessed, as well as anthropometric and biochemical blood profiles. In addition, SAT biopsies were performed for histologic and morphometric analyses. Statistical analysis was carried out with R Studio, using descriptive and inferential analysis, ANOVA test, and mixed-effect model (P < 0.05). HIV+ showed higher levels of very-low-density lipoproteins and triglycerides and lower levels of high-density lipoproteins at baseline than HIV– (P < 0.05). All groups showed improved aerobic and strength performances (P < 0.05). Both groups showed reduced adipocyte sizes after CT (P < 0.05). Lastly, HIV+ presented smaller adipocytes and higher elastic fiber deposition at baseline and decreased after training only in HIV+, similar to the HIV group. Thus, CT in PLWHA promoted a decrease in the size heterogeneity of adipocytes and elastic fiber deposition, remodeling the ECM, and improving the SAT fibrosis profile. Brazilian Clinical Trials Registry (ensaiosclinicos.gov.br – UTN: U1111-1214-3022).Novelty: Adipose tissue fibrosis is improved by training in people living with HIV. Concurrent training remodels adipose tissue extracellular matrix.

The aim of this work was to evaluate the effects of sintering temperature over the structural, physical and mechanical properties in addition to the in vivo biological performance of nanostructured biocomposites obtained from the mixture between calcined fish bone (hydroxyapatite + β-tricalcium phosphate) and niobium pentoxide. The sintering temperatures varied from 650 to 1150°C, in steps of approximately 50°C. The results showed the formation of a stable triphasic nanostructured bioceramic with almost constant relative proportions between the CaNb2O6, PNb9O25 and Ca3(PO4)2 phases. Special attention was given to the sintering temperature at 1080°C because there was occurrence of a liquid phase, and the sample showed localised maximisations of linear shrinkage, density and Vickers hardness. This niobium-biphasic calcium phosphate 1080 (Nb-BCP-1080) nanostructured biocomposite presented measured values for compressive strength (242 ± 29 MPa), Young's modulus (19.63 ± 3.5 GPa), Poisson's ratio (0.248 ± 0.016) and flexural strength (24 ± 5.9 GPa) close to those of some human bones. Then, to further explore these characteristics, in vivo studies were performed by implanting 8.0-mm diameter discs of this material in calvaria of rats. Histological observation showed the occurrence of osseointegration between the neo-formed bone and the triphasic bioceramic just 45 days after implantation. In conclusion, the observations of this work unambiguously show that CaNb2O6–PNb9O25–Ca3(PO4)2 triphasic bioceramics are candidates for bone replacements where high and medium loads are demanded.

The search for gold-standard materials for bone regeneration is still a challenge in reconstruction surgery. The ratio between hydroxyapatite (HAp) and β-tricalcium phosphate (β-TCP) in biphasic calcium phosphate ceramics (BCPs) is one of the most important factors in osteoinduction promotion and controlled biodegradability, configurating what is currently considered as a possible gold standard material for bone substitution in reconstructive surgery. Exploring the natural genesis of the HAp and β-TCP phases in fishbones during their postnatal growth, this study developed a biphasic bioceramic obtained from the calcination of Nile tilapia (Oreochromis niloticus) bones as a function of their ages. The natural genesis dynamics of the structural evolution of the β-TCP and HAp phases were characterized by physicochemical methods, taking into account of the age of the fish and the material processing conditions. Thermal analysis (TGA / DTA) showed complete removal of the organic matter and transitions associated with the transformation of carbonated hydroxyapatite (CDHA) to HAp and β-TCP phases. After calcination at 900 °C, the material was characterized by: X-ray diffraction (XRD) and refinement by the Rietveld method; Fourier Transform Infrared Spectroscopy with Attenuated Total Reflection (FTIR-ATR); Raman spectroscopy; Scanning Electron Microscopy (SEM) and Flame Atomic Absorption Spectroscopy (FAAS). The analysis allowed identification and quantitative estimate of the variations of the HAp and β-TCP phases in the formation of the BCPs. The results showed that the decrease in β-TCP against the increase in the HAp phases is symmetrical to the dynamics of the natural genesis of these phases, surprisingly maintaining the balanced phase proportion even when bones of young fishes were used. The microstructure analysis confirms the observed transformation. In addition, in vivo tests demonstrated the osteoinductive potential of BCP scaffolds implanted in an ectopic site, and their remarkable regenerative functionality, as bone graft, was demonstrated in alveolar bone after tooth extraction. MTT cytotoxicity assay for BCP samples for MC3T3-E1 pre-osteoblasts and L929 fibroblasts cells showed viability equal or higher than 100%. A logistic empirical model is presented to explain the three stages of HAp natural formation with fish age and it is also compared to the fish size evolution.

In this work, the application of photoacoustic and photothermal methods for the evaluation of physicochemical properties of biological systems will be presented as well as the state of art of these methods fostering clinical diagnostics.

Fish bones are a natural calcium phosphate (CaP) sources used in biomaterials production for bone regeneration. CaP scaffolds can be enriched with other substances with biological activity to improve bone repair. This study aimed to evaluate the physicochemical properties and bone regeneration potential of biphasic calcium phosphate (BCP) scaffolds impregnated with free curcumin (BCP-CL) or complexed with β-cyclodextrin (BCP-CD) compared to BCP scaffolds. Rietveld’s refinement showed that BCP is composed of 57.2% of HAp and 42.8% of β-TCP and the molar ratio of Ca/P corresponds to 1.59. The scaffolds presented porosity (macro and microporosity) of 57.21%. Apatite formation occurred on the BCP, BCP-CL, and BCP-CD surface, in vitro, in SBF. Micro-Raman technique showed a reduction in the dissolution rate of β-TCP in the curcumin-impregnated scaffolds over time, and in vivo studies on critical-size defects, in rat calvaria, had no additional regenerative effect of BCP-CL and BCP-CD scaffolds, compared to BCP scaffolds. Despite this, the study showed that curcumin impregnation in BCP scaffolds prolongs the release of the β-TCP phase, the BCP- phase with the higher osteoinductive potential, representing an advantage in tissue engineering.

Natural or synthetic biomaterials are increasingly being used to support bone tissue repair or substitution. The combination of natural calcium phosphates with biocompatible alloys is an important route towards the development of new biomaterials with bioperformance and mechanical responses to mimic those of human bones. This article evaluated the structural, physical, mechanical and biological properties of a new mechanical improved nanocomposite elaborated by association of fish biphasic calcium phosphate (BCP) and niobium pentoxide (Nb2O5). The nanocomposite (Nb-BCP) and the pure BCP, used as a positive control, were obtained by powder metallurgy. The density, porosity and microhardness were measured. The structural analysis was determined by X-ray diffraction (XRD) and the biological properties were studied in histological sections of critical size calvaria defects in rats, 7, 15, 30, 45 and 60 days after implantation of disks of both materials. Morphological description was made after scanning electron microscopy (SEM) and optical microscopy analysis. After sintering, the Nb-BCP nanocomposite presented four crystalline phases: 34.36% calcium niobate (CaNb2O6), 21.68% phosphorus niobium oxide (PNb9O25), 42.55% β-tricalcium phosphate (Ca3(PO4)2) and 1.31% of niobium pentoxide (Nb2O5) and exhibited increases of 17% in density, 66% in Vickers microhardness and 180% in compressive strength compared to pure BCP. In vivo study, showed biocompatibility, bioactivity and osteoconductivity similar to pure BCP. SEM showed the formation of globular accretions over the implanted nanocomposites, representing one of the stages of bone mineralization. In conclusion, the BCP and Nb2O5 formed a nanocomposite exhibiting characteristics that are desirable for a biomaterial, such as bioperformance, higher β-TCP percentage and improved physical and mechanical properties compared to pure BCP. These characteristics demonstrate the promise of this material for supporting bone regeneration.

Aerobic exercise training (AER) may promote several adaptations in white adipose tissue (WAT), including a phenotypic change known as browning. The present study aimed at assessing if resistance exercise training (RES) would be as efficient as AER in inducing a brown-like adipocyte reprogramming in WAT. Thirty Swiss male mice were randomly divided into 3 groups with 10 animals each: 1) sedentary (SED), 2) AER, and 3) RES. After the adaptation training, an incremental test was performed at the beginning of each week to adjust training load. Mice were submitted to 8 wk of AER or RES. After the experimental period, inguinal and retroperitoneal WAT (iWAT and rpWAT) and brown adipose tissue (BAT) were collected. The prescription of AER and RES was effective in increasing the performance of both groups. Also, RES presented a lower body weight than AER/SED. AER and RES reduced the area of iWAT and rpWAT adipocytes and the lipid area of BAT, induced an increase of vascular endothelial growth factor (VEGF) and cluster of differentiation 31 (CD31) and uncoupling protein 1 (UCP-1), and increased the expression of selective genes of brown and beige phenotype in adipocytes after 8 wk. In general, we demonstrated here that AER and RES training similarly induced the browning of iWAT and rpWAT.NEW & NOTEWORTHY Aerobic exercise training (AER) induces the browning of white adipose tissue, turning adipocytes multilocular, highly vascularized and expressing uncoupling protein 1 (UCP-1). The current study compared the efficiency of resistance to aerobic exercise training to promote a brown-like phenotype. Our results suggest that both types of training similarly induce subcutaneous and visceral adipose tissue browning.

Natural or synthetic biomaterials are increasingly being used to support bone tissue repair or substitution. The combination of natural calcium phosphates with biocompatible alloys is an important route towards the development of new biomaterials with bioperformance and mechanical responses to mimic those of human bones. This article evaluated the structural, physical, mechanical and biological properties of a new mechanical improved nanocomposite elaborated by association of fish biphasic calcium phosphate (BCP) and niobium pentoxide (Nb

Aim: To evaluate if radiation used in radiotherapy can cause changes in the virulence potential of Candida tropicalis ATCC 750. Materials & methods: C. tropicalis was exposed in vitro to identical dose and scheme of irradiation would be used in patients with head and neck cancer. Some virulence parameters were analyzed before and after irradiation. Results: Colony morphologies were irreversibly affected by irradiation. Increase in growth rate, filamentation, adhesion on cell lines and phagocytosis process were also observed. Overall the irradiated C. tropicalis cells became more efficient at causing systemic infection in mice. Conclusion: γ-radiation induced important changes in C. tropicalis increasing its virulence profile, which could directly affect the relationship between yeasts and hosts.

Studies have shown that radiation from radiotherapy increases the yeast colonization of patients. However it is not clear, if such radiation alters the yeast itself. The aim of the present study was therefore to report the direct impact of gamma radiation on Candida tropicalis. C. tropicalis was obtained from a patient with a carcinoma, a suspension of this yeast containing 2.0 × 103 colony forming units per milliliter was prepared. It was submitted to gamma radiation dosage similar to that used in the treatment of head and neck cancer. After a cumulative dose of 7200 cGy some virulence attributes of C. tropicalis, including macro and micromorphological characteristics, adhesion and biofilm abilities, murine experimental infection and phagocytosis resistance were evaluated on irradiated and non-irradiated yeasts. After irradiation the colony morphology of the yeast was altered from a ring format to a smooth appearance in most colonies. Scanning electron microscopy revealed notable differences in the structures of both these colonies and the yeast cells, with the loss of pseudohyphae following irradiation and an increase in extracellular matrix production. The adherence and biofilm production of the yeast was greater following irradiation, both in terms of the number of yeasts and total biomass production on several abiotic surfaces and TR146 cells. The phagocytic index of the irradiated yeasts was not statistically different; however, the presence of cellular debris was detected in the kidneys of infected animals. Mice infected with irradiated yeasts developed an infection at the site of the yeast inoculation, although systemic infection was unchanged. Our findings show for the first time that C. tropicalis, one of the most important yeasts from colonization, which cause fatal candidemia in cancer patients, is affected by gamma irradiation, with changes to its virulence profile.

Nowadays the increase in life expectancy of the population has made diseases related to bone structure a worldwide public health problem. In the field of bone regeneration, calcium phosphate-based ceramics hold the greatest promise of efficiency and hydroxyapatite presents itself as reference material because, in addition to presenting a chemical composition similar to the mineral composition of bone tissue, it is biocompatible, osteoconductive, and osteoinductive, and can be reinforced with other more resistant materials, such as titanium, aluminum, zirconia, and niobium. In this scenario, fish bones represent an abundant and low-cost natural source of hydroxyapatite that is promising for the production of more resistant devices for bone regeneration. The objective of this work was to evaluate the physicochemical properties of the natural HAp, extracted from bones of Nile tilapia (Oreochromis niloticus) and of the HAp-Nb-1080 composite. Besides, the bioperformance of scaffoldings of both materials was evaluated in terms of bone regeneration. The granulometric, XRD, ATR-FTIR, micro-Raman, micro-CT and, SEM analyzes were performed to HAp and HAp-Nb-1080 powders characterization. The bone regeneration was studied 15, 30, and 45 days after implantation of 8 mm scaffolds in critical-size defects in rat calvaria. The results showed that fishbone HAp presented the molar ratio (Ca/P) of 1.66. The average particle sizes of the HAp and HAp-Nb-1080 powders were in the order of 1150 and 750 nm, respectively. After sintering the composite, three new crystalline phases were formed: β-tricalcium phosphate Ca3(PO4)2 (34%), fersmite (CaNb2O6) (33.5%), and phosphorus niobium oxide (PNb9O25) (32.6%), confirmed by ATR-FTIR, and micro-Raman analyzes. The scaffolds porosities were 57% to HAp and 59% to HAp-Nb-1080 composite. SEM results showed the presence of opened macro and microporosity as well as porous interconnectivity. The histological study showed bone formation inside porous in all analyzed time interval after the implantation. In this study, we demonstrated that the HAp-Nb-1080 composite presented physicochemical characteristics that satisfy the requirements for a mechanically reinforced biomaterial for bone regeneration.

Paracoccidioides brasiliensis (Pb) can disseminate through the lymphatic and hematogenic pathways. As a result the spleen and other lymphoid organs are targets of paracoccidioidomycosis. There are few studies describing this disease in the spleen and more detailed descriptions are required. This study combines Histopathology and Fourier Transform Raman Spectroscopy (FT-Raman) methods to study spleen infected by Pb. The Swiss mice were euthanized after 1, 2, 4 and 8 weeks of infection with Pb, and their spleens were removed for ex vivo analyzes. Histopathological evaluation revealed that the red pulp was the area most affected, presenting the highest concentration of yeasts, inflammatory cells and cells in apoptosis, with no observation of typical granuloma formation. The main physical-chemical changes were detected in the spectral ranges between 1730-1590 cm-1 and 1390-1280 cm-1 attributed to amide I and vibrational deformation mode of CH3 and CH2 molecules, respectively. The amide I was correlated to total protein content and CH3 and CH2 molecules to collagen of the spleen. The association between histological and physical-chemical methods enabled detection of several alterations in the spleen, including apoptosis, contributing to a better understanding of paracoccidioidomycosis, a tropical neglected disease.

Recent evidence includes apoptosis as a defense against Trypanosoma cruzi infection, which promotes an immune response in the host induced by T cells, type 1, 2 and 17. Currently, there is no medicine completely preventing the progression of this disease. We investigated the immunological and apoptotic effects, morbidity and survival of mice infected with T. cruzi and treated with dynamized homeopathic compounds 13c: Kalium causticum (GCaus), Conium maculatum, (GCon), Lycopodium clavatum (GLy) and 7% alcohol solution (control, vehicle compounds, GCI). There was significant difference in the increase of apoptosis in the treated groups, compared with GCI, which might indicate action of the compounds in these cells. Infected animals treated with Lycopodium clavatum presented better performance compared with other groups. GLy showed a higher amount of hepatocytes and splenocytes undergoing apoptosis, higher number of apoptotic bodies in the liver, predominance of Th1 response, increased TNF-α and decreased IL-6, higher survival, lower morbidity, higher water consumption, body temperature, tendency to higher feed intake and weight gain compared with GCI. Conium maculatum had worse results with increased Th2 response with increased IL-4, worsening of the infection with early mortality of the animals. Together, these data suggest that highly diluted medicines modulate the immune response and apoptosis, affecting the morbidity of animals infected with a highly virulent strain of T. cruzi, being able to minimize the course of infection, providing more alternative approaches in the treatment of Chagas disease.

This study evaluated whether anethole attenuates the inflammatory response and joint damage in a model of adjuvant-induced arthritis (AIA) in rats. The animals were treated with 62.5-, 125-, or 250-mg/kg anethole daily for 21 days after AIA and necropsied on days 14 and 21 to evaluate the number of serum and synovial leukocytes (total and differential), serum cytokines (IL-2, IL-6, IL-12, IL-17, and TNF-α), and nitric oxide concentrations. Morphologic changes in the cartilage and bone of the femorotibial articulation in both left paw and right paw were studied in hematoxylin/eosin and Sirius Red-hematoxylin sections. Different doses of anethole suppressed paw swelling and the number of serum and synovial leukocytes. However, 250 mg/kg of anethole more effectively controlled local and systemic inflammation. Histological evaluation revealed significant prevention of cartilage damage and inflammatory infiltrate scores. Morphometric analysis showed pannus formation, the thickness of the articular cartilage, and bone resorption lower in the anethole-treated AIA group compared to untreated AIA group on both days 14 and 21. These significant anti-inflammatory effects in the anethole-treated AIA group were associated with downregulation of cytokines and nitric oxide levels. Therefore, anethole may be a useful intervention to treat inflammatory arthritis.

In this study was investigated the effect of treatment with a fish oil preparation (FOP) on inflammatory and histopathological parameters in a rat model of adjuvant-induced arthritis (AIA). Paw edema, the severity of secondary lesions, the number of synovial leukocytes, and serum nitric oxide and cytokines concentrations were evaluated. Histological changes in cartilage and bone of the femorotibial articulation in the paws were analyzed by hematoxylin/eosin and sirius red-hematoxylin staining. Oral treatment with the FOP (75, 150, and 300 mg/kg) that was initiated on the day of arthritis induction (day zero) and maintained for 21 days inhibited inflammatory signals and prevented morphological changes in cartilage and bone of the femorotibial articulation of the paws. These effects could be at least partially attributable to inhibitory and modulatory actions of the FOP on the production and release of nitric oxide and cytokines that participate in the pathophysiology of AIA. En este estudio se investigó el efecto del tratamiento con una preparación de aceite de pescado (PAP) sobre los parámetros inflamatorios y histopatológicos en un modelo de artritis inducida por adyuvante en ratas (AIA). Se evaluó el edema de la pata, la gravedad de las lesiones secundarias, el número de leucocitos sinoviales y las concentraciones séricas de óxido nítrico y citocinas. Los cambios histológicos en el cartílago y el hueso de la articulación femorotibial en las patas se analizaron mediante tinción con hematoxilina / eosina y rojo de sirio-hematoxilina. El tratamiento oral con PAP (75, 150 y 300 mg / kg) que se inició el día de la inducción de la artritis (día zero) y se mantuvo durante 21 días inhibió las señales inflamatorios y previno cambios morfológicos en el cartílago y hueso de la articulación femorotibial de las patas. Estos efectos podrían atribuirse, al menos en parte, a las acciones inhibidoras y moduladoras de la PAP sobre la producción y liberación de óxido nítrico y citocinas que participan en la fisiopatología de la AIA. Nesse estudo foi investigado o efeito do tratamento com uma preparação de óleo de peixe (POP) sobre parâmetros inflamatórios e histopatológicos na artrite induzida por adjuvante em ratos (AIA). O edema de pata, a gravidade das lesões secundárias, o número de leucócitos sinoviais e a concentração sérica de óxido nítrico e de citocinas foram avaliados. Alterações histológicas nas cartilagens e nos ossos das articulações fêmoro-tibiais das patas foram analisadas pelas técnicas de coloração hematoxilina/eosina e sirius red-hematoxilina. O tratamento oral com a POP (75, 150, e 300 mg/kg) iniciado no dia da indução da artrite (dia zero) e mantido por 21 dias inibiu os sinais inflamatórios e preveniu as alterações morfológicas na cartilagem e no osso das articulações. Esses efeitos podem ser parcialmente atribuídos à ação modulatória e inibitória da POP sobre a produção/liberação de óxido nítrico e citocinas que participam da fisiopatologia da AIA.

The secondary metabolites produced by Fusarium can cause disease and death when consumed and produce biological responses even in the absence of the microorganism. The IL-6, TNF-α and TGF-β1 cytokines immune reactivity was associated with histopathological and physico-chemical changes in skin of immune competent rats after administration of Fusarium oxysporum crude extract. Rats were intradermally injected with 50 μl of 0.5 mg/ml crude extract and were euthanized at 3, 6, 12 and 24 h after injection. The inflammatory response was quantified by enzyme myeloperoxidase activity and by immunohistochemical method to detect the IL-6, TNF-α and TGF-β1. Physico-chemical analysis was performed using FT-Raman Spectroscopy. The inflammatory response was most intense at 6 and 12 h after crude extract administration and the most significant histopathological changes were observed in the dermis. Myeloperoxidase activity was intense from 3 to 24 h after injection. The immunostaining of pro-inflammatory cytokines IL-6 and TNF-α peaked at 6 h. Immunostaining for TGF-β1 was highest at 12 and 24 h. FT-Raman spectral analysis showed both, the most intense Fusarium interaction with the skin at 6 h, as revealed by the changes in the stretching of –CH bands (3100–2800 cm−1) in the dermis, and skin recovery trending after 12 h after crude extract injection. The results showed that secondary metabolites stimulated histopathologic changes and inflammatory responses even in the absence of the fungus, increasing myeloperoxidase activity and pro-inflammatory cytokine expression besides promoting physico-chemical changes.

Guided Bone Regeneration (GBR) is a technique based on the use of a physical barrier that isolates the region of bone regeneration from adjacent tissues. The objective of this study was to compare GBR, adopting a critical-size defect model in rat calvaria and using collagen membrane separately combined with two filling materials, each having different resorption rates. A circular defect 8 mm in diameter was made in the calvaria of Wistar rats. The defects were then filled with calcium sulfate (CaS group) or deproteinized bovine bone mineral (DBBM group) and covered by resorbable collagen membrane. The animals were killed 15, 30, 45 and 60 days after the surgical procedure. Samples were collected, fixed in 4% paraformaldehyde and processed for paraffin embedding. The resultant sections were stained with H&E for histological and histomorphometric study. For the histomorphometric study, the area of membrane was quantified along with the amount of bone formed in the region of the membrane. Calcium sulfate was reabsorbed more rapidly compared to DBBM. The CaS group had the highest percentages of remaining membrane at 15, 30, 45 and 60 days, compared to the DBBM group. The DBBM group had the highest amount of new bone at 45 and 60 days compared to the CaS group. Based on these results, it was concluded that the type of filling material may influence both the resorption of collagen membrane and amount of bone formed.

The purpose of this study is to evaluate the toxicity of synthetic chalcones 1 and 2 in uninfected hamsters and anti-Leishmania activity of synthetic chalcones 1 and 2 in hamsters infected with Leishmania (Viannia) braziliensis. For the toxicity test, uninfected animals were treated with chalcones 1 and 2, and clinical and biochemical parameters and histological aspects of the liver and kidneys were assessed. Chalcones 1 and 2 were then intraperitoneally or topically administered (10 mg/kg body weight) three times per week in animals infected with promastigotes of L. (V.) braziliensis. We monitored the thickness of the infected footpads, determined parasitic load, performed histological analysis, and detected apoptosis in situ. The results were analyzed using Student’s t test and Mann-Whitney test at a significance level of 5 %. Neither of the chalcones showed toxicity. Chalcone 2 administered intraperitoneally significantly reduced the thickness of the infected footpad compared with the beginning of treatment. The parasite load of the lymph node and spleen was reduced in the groups treated with chalcones 1 (topical) and 2 (intraperitoneal). Chalcone 2 (topical) reduced parasite burden only in the lymph node. The histological analysis revealed reconstitution of the tissue and reductions of inflammation and apoptosis in the infected footpad in these groups. The synthetic chalcones 1 (topical) and 2 (intraperitoneal and topical) at a dose of 10 mg/kg showed anti-Leishmania activity in vivo, no renal or hepatic toxicity, and a reduction of apoptosis of the cells in the lesions. These chalcones may have substantial potential for the treatment of cutaneous leishmaniasis.

Oxidative stress is an important factor modulating skin alterations. Melochia arenosa Benth. (Malvaceae) is a Brazilian plant with antimicrobial activity and antioxidant potential.The objective of this study is to develop a topical formulation containing antioxidant phenolic-rich extract of M. arenosa and to evaluate its skin permeation profile.Response surface methodology was used to maximize the total phenolic (TP) content of the extract and its antioxidant activity was evaluated by 2,2-diphenyl-1-picryl-hydrazyl (DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and respiratory burst methods. An emulsion containing 1% optimized extract (OE) was developed and employed photoacoustic spectroscopy (PAS) for the determination of its skin permeation profile. The morphology of the skin was studied in histological sections stained with hematoxylin-eosin.The optimum conditions predicted for the major extractive efficiency of the phenolics with 100% ethanol led extraction time 101 h and plant:solvent proportion 1:13.5 (w/v). OE presented TP = 724.6 ± 8.2 mg GAE/g extract and scavenging capacity of DPPH (IC50 value = 11.43 ± 0.14 µg/mL) and ABTS radicals (IC50 value = 35.42 ± 0.48 µg/mL). The production of ROS by neutrophils after stimulation with phorbol miristate acetate was lower when the OE was present in the reaction medium, endorsing its high antioxidant capacity. The data obtained by PAS indicated that the OE present in the emulsion has permeated and was distributed in the whole skin. No histopathological alterations were observed in the histological analysis.The formulation developed is a promising tool for skin care and could prevent the damage caused by oxidative stress.

Objective : The aim of this study was make a descriptively evaluate the repair of the periapical region of dog teeth. Material and Methods : Thirty-six root canals were used, eighteen were filled with 0.075% capsaicin and calcium hydroxide (test paste) and the rest eighteen root canals were filled with calcium hydroxide (control group). Histological analyzes were performed to classify inflammation as absent, mild, moderate or severe. The analyses also classified the presence or absence of repair at 7, 14 and 30 days. Results : The use of 0.075% capsaicin did not modify the inflammatory response and repair when compared to control group. Conclusion : Our data suggests that test group with 0.075% capsaicin as intracanal administration could be indicated for tissue repair; furthermore other experiments should be carried out to evaluate possible clinical use of capsaicin for atypical odontalgia. Keywords : Capsaicin; Repair; Inflammation; Periapical tissue.

This study evaluates the effect of Trypanosoma cruzi biotherapy 17dH (BIOT) on mice of different ages, infected with the protozoa concerned.Performing a blind, controlled, randomized by drawing experiment, 110 animals four or eight-week-old, Swiss, male mice were divided into infected control treated hydroalcoholic 7% (CI-4 = 34 or CI-8 = 21 animals) and infected control treated with biotherapy 17dH-0.2 mL/animal/20 consecutive days/oral regimen (BIOT-4 = 33 or BIOT-8 = 21 animals). Animals were inoculated intraperitoneally with 1400 trypomastigote, T. cruzi Y-strain. Parasitological, immunological and histopathologic parameters were evaluated statistically, using Statistica-8.0 and R 3.0.2 program to analysis of survival. The study was approved by the Ethics Committee for Animal Experimentation/UEM.Four-week-old mice showed no statistical difference in parasitemia (P = 0.5718) between the treated and control group. Eight-week-old mice from the treated group had a higher parasite peak (P = 0.0424) and higher parasitemia (P 0.005) than the control. To both groups of 4 and 8 weeks of age, treated or untreated, survival of mice was higher in the treated group than in the control, although it was not statistically significant (p-value = 0.32, 0.55 respectively). Four-week-old mice displayed a spleen section with a number of amastigote nests significantly higher in BIOT-4 than CI-4 (P = 0.01). In eight-week-old mice the number of amastigote nests (P 0.001) and inflammatory foci (P 0.06-10% significance) in the liver section were smaller in BIOT-8 than CI-8. Spleen giant cells were significantly higher in CI-8 than in BIOT-8 (P 0.01). Eight-week-old animals treated with biotherapy showed higher parasitemia and lower tissue parasitism. Opposite pattern was observed in four-week-old animals.There is a difference of high diluted medication effect in four and eight-week-old mice. In the group of animals 8 weeks the immunomodulatory effect seems to have been higher. Hence, treatment with the medicine produced from T. cruzi modulates the inflammatory response with increased apoptosis and decreased serum levels of TGF-β.

OBJECTIVE: To evaluate the effect of two different concentrations of sodium alendronate on the quantity of organic matrix in the femur of rats with estrogen suppression caused by ovariectomy.METHODS: Sixty-days-old Wistar rats (Rattus norvegicus) were subjected to bilateral laparotomy to remove the ovaries. The animals were divided into a control group, in which they only underwent laparotomy; an ovariectomized group (OVX); an ovariectomized group treated with 1 mg/kg of alendronate (OVX 1 mg); and an ovariectomized group treated with 2 mg/kg of alendronate (OVX 2 mg). The rats received alendronate twice a week for 90 days. The left femur was then removed, fixed and processed for embedding in paraffin. Semi-serial sections stained with hematoxylin and eosin were used to determine the area occupied by organic bone matrix, by means of image analysis software. The animals' weights were obtained at the beginning and end of the experiment.RESULTS: The ovariectomized animals and those treated with 1 mg/kg of alendronate presented significant increases in body weight (p < 0.05), in comparison with the control group. Histomorphometric analysis revealed that in the animals treated with 2 mg/kg of alendronate, the area (µm2) occupied by organic matrix (1,81,900 ± 18,130) was similar (p > 0.05) to that of the non-ovariectomized control animals (2,04,800 ± 9590), which indicates that this medication had a preventive effect with regard to bone mass loss.CONCLUSION: The higher concentration of the medication, administered twice a week for 90 days, was more effective than the dose of 1 mg/kg over the same period. OBJETIVO: avaliar a efeito de duas concentrações diferentes de alendronato de sódio (ALN) sobre a quantidade de matriz orgânica no fêmur de ratas com supressão estrogênica provocada por ovariectomia.MÉTODOS: ratas Wistar (Rattus norvegicus) com 60 dias foram submetidas a laparotomia bilateral para remoção dos ovários. Os animais foram divididos em grupo controle, no qual os animais foram apenas laparotomizados; grupo ovariectomizado (OVX); grupo ovariectomizado tratado com 1 mg/kg de ALN (OVX 1 mg); e grupo ovariectomizado tratado com 2 mg/kg de ALN (OVX 2 mg). Receberam ALN duas vezes por semana durante 90 dias. O fêmur esquerdo foi coletado, fixado e processado para inclusão em parafina. Cortes semisseriados corados com H&E foram usados para a obtenção, com um software de análise de imagens, da área ocupada por matriz óssea orgânica. O peso dos animais foi obtido no inicio e no fim do experimento.RESULTADOS: os animais ovariectomizados e aqueles tratados com 1 mg/kg de ALN tiveram um aumento significativo (p < 0,05) no peso corporal quando comparados com o grupo controle. A análise histomorfométrica revelou que nos animais tratados com 2 mg/kg de ALN a área (µm2) ocupada por matriz orgânica (181.900 ± 18.130) foi semelhante (p > 0,05) àquela dos animais controle não ovariectomizados (204.800 ± 9.590), o que indica um efeito preventivo desse medicamento sobre a perda de massa óssea.CONCLUSÃO: a maior concentração do medicamento administrado duas vezes por semana por 90 dias foi mais eficaz do que a dose de 1 mg/kg no mesmo período.

ResumoObjetivoavaliar a efeito de duas concentrações diferentes de alendronato de sódio (ALN) sobre a quantidade de matriz orgânica no fêmur de ratas com supressão estrogênica provocada por ovariectomia.Métodosratas Wistar (Rattus norvegicus) com 60 dias foram submetidas a laparotomia bilateral para remoção dos ovários. Os animais foram divididos em grupo controle, no qual os animais foram apenas laparotomizados; grupo ovariectomizado (OVX); grupo ovariectomizado tratado com 1 mg/kg de ALN (OVX 1 mg); e grupo ovariectomizado tratado com 2 mg/kg de ALN (OVX 2 mg). Receberam ALN duas vezes por semana durante 90 dias. O fêmur esquerdo foi coletado, fixado e processado para inclusão em parafina. Cortes semisseriados corados com H&E foram usados para a obtenção, com um software de análise de imagens, da área ocupada por matriz óssea orgânica. O peso dos animais foi obtido no inicio e no fim do experimento.Resultadosos animais ovariectomizados e aqueles tratados com 1mg/kg de ALN tiveram um aumento significativo (p0,05) àquela dos animais controle não ovariectomizados (204.800±9.590), o que indica um efeito preventivo desse medicamento sobre a perda de massa óssea.Conclusãoa maior concentração do medicamento administrado duas vezes por semana por 90 dias foi mais eficaz do que a dose de 1mg/kg no mesmo período.AbstractObjectiveto evaluate the effect of two different concentrations of sodium alendronate on the quantity of organic matrix in the femur of rats with estrogen suppression caused by ovariectomy.Methodssixty‐day‐old Wistar rats (Rattus norvegicus) were subjected to bilateral laparotomy to remove the ovaries. The animals were divided into a control group, in which they only underwent laparotomy; an ovariectomized group (OVX); an ovariectomized group treated with 1 mg/kg of alendronate (OVX 1 mg); and an ovariectomized group treated with 2 mg/kg of alendronate (OVX 2 mg). The rats received alendronate twice a week for 90 days. The left femur was then removed, fixed and processed for embedding in paraffin. Semi‐serial sections stained with hematoxylin and eosin were used to determine the area occupied by organic bone matrix, by means of image analysis software. The animals’ weights were obtained at the beginning and end of the experiment.Resultsthe ovariectomized animals and those treated with 1mg/kg of alendronate presented significant increases in body weight (p0.05) to that of the non‐ovariectomized control animals (204,800±9,590), which indicates that this medication had a preventive effect with regard to bone mass loss.Conclusionthe higher concentration of the medication, administered twice a week for 90 days, was more effective than the dose of 1mg/kg over the same period.

The skin is an important gateway for Fusarium infection in humans. Our hypothesis is that metabolites produced by Fusarium oxysporum should change the barrier structure to permeate the skin. Male Wistar rats received a topical application of a solution (0.05 mg/mL) of Fusarium metabolites. The animals were euthanized 3, 6, 12, 24 h after and the skin was processed for immunostaining by laminin and E-cadherin to investigate whether the Fusarium metabolites can break the barrier of healthy skin. Other techniques were employed: HE to study the morphology; metalloproteinase-9 (MMP-9), TUNEL, and PCNA immunostaining to evaluate the inflammation, cell death, and proliferation, respectively. There was an inflammatory response mainly centered in the dermis. Qualitatively, the skin of the experimental group showed reduced E-cadherin and laminin immunostaining at 3, 12, and 24 h. Higher intensity staining by TUNEL at 3 h, and PCNA at 6, 12, and 24 h. There was intense MMP-9 activity at 6, 12, and 24 h. None of analyses revealed any changes in the epidermis. It was concluded that the fraction was able to permeate the skin and act selectively in dermis, inducing inflammatory response, increasing MMP-9 immunostaining, inducing apoptosis, and reducing E-cadherin and laminin immunostaining.

Fusarium is an emerging pathogen with high levels of morbidity and mortality. A significant tissue response is observed in infected patients, and the condition has been associated with the production of toxic metabolites. The aim of the present study was to identify a major fraction of crude metabolic extract of Fusarium oxysporum and investigate its effects on the skin of healthy rats. Fraction F1 was obtained from the cultivation of F. oxysporum in Czapek–Dox. In the treatment groups, fraction F1 (0.05 mg/ ml) was injected intradermally, while (50 µl) 0.9% of saline solution was injected in the control groups. The animals were killed 3, 6, 12, and 24 h after inoculation. The skin was fixed for inclusion in paraffin to obtain histological sections and stained with hematoxylin and eosin, Sirius red, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. Samples were analyzed using Fourier transform Raman spectroscopy. The tissue reactions were classified and compared over time and by treatment. In the treatment group, inflammatory reaction peaked at 6 h, being classified as moderate, with infiltrate composed mainly of neutrophils. terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining was negative. The area occupied by types I and III collagen in the treatment group increased over time. There was a change in the area occupied by amide I and the ratios of the –CH2 and –CH3 molecules. It can be argued that the fraction F1 contains elements that contribute to the invasion of Fusarium in the skin, destructurizing the organization of the extracellular matrix. Copyright © 2014 John Wiley & Sons, Ltd.

The cleft palate presented by transforming growth factor-β3 (Tgf-β3) null mutant mice is caused by altered palatal shelf adhesion, cell proliferation, epithelial-to-mesenchymal transformation and cell death. The expression of epidermal growth factor (EGF), transforming growth factor-β1 (Tgf-β1) and muscle segment homeobox-1 (Msx-1) is modified in the palates of these knockout mice, and the cell proliferation defect is caused by the change in EGF expression. In this study, we aimed to determine whether this change in EGF expression has any effect on the other mechanisms altered in Tgf-β3 knockout mouse palates. We tested the effect of inhibiting EGF activity in vitro in the knockout palates via the addition of Tyrphostin AG 1478. We also investigated possible interactions between EGF, Tgf-β1 and Msx-1 in Tgf-β3 null mouse palate cultures. The results show that the inhibition of EGF activity in Tgf-β3 null mouse palate cultures improves palatal shelf adhesion and fusion, with a particular effect on cell death, and restores the normal distribution pattern of Msx-1 in the palatal mesenchyme. Inhibition of TGF-β1 does not affect either EGF or Msx-1 expression.

This study evaluated physical, chemical and morphological changes in lungs of mice infected with Paracoccidioides brasiliensis. The animals were inoculated with 0.1 mL of fungal suspension of the P. brasiliensis 18 isolate and were euthanized 1, 2, 4 and 8 weeks after inoculation. The upper left lobe of the lung was isolated, fixed and processed for paraffin embedding. The sections were stained with H&E for histopathological study, with Gomori-Grocott to locate and identify the fungus, and with TUNEL immunostaining to detect the occurrence of programmed cell death. The lower and middle right lobes were analyzed by Fourier Transform Infrared Photoacoustic Spectrocopy (FTIR-PAS) to investigate physical and chemical features of the infected lungs. The results showed that lungs infected by P. brasiliensis underwent structural changes that varied according to the time period analyzed, and that changes in the absorption bands of different chemical groups resulted from these morphological changes. The results suggest that the combination of FTIR-PAS spectroscopy with morphological evaluation is an effective procedure for the study of paracoccidioidomycosis, one of the most important systemic mycoses that can damage the lung architecture and consequently impair the respiratory function.

Creatine is widely used by athletes as an ergogenic resource. The aim of this study was to evaluate the infl uence of creatine supplementation on the duodenum of rats submitted to physical training. The number and myenteric neuronal cell bodies as well mucosal and muscular tunic morphometry were evaluated. Control animals received a standard chow for 8 weeks, and the treated ones received the standard chow for 4 weeks and were later fed with the same chow but added with 2% creatine. Animals were divided in groups: sedentary, sedentary supplemented with creatine, trained and trained supplemented with creatine. The training consisted in treadmill running for 8 weeks. Duodenal samples were either processed for whole mount preparations or for paraffi n embedding and hematoxylin-eosin staining for histological and morphometric studies of the mucosa, the muscular tunic and myenteric neurons. It was observed that neither creatine nor physical training alone promoted alterations in muscular tunic thickness, villus height or crypts depth, however, a reduction in these parameters was observed when both were associated. The number of myenteric neurons was unchanged, but the neuronal cell body area was reduced in trained animals but not when training and creatine was associated, suggesting a neuroprotector role of this substance

Introduction Skin lesions are a significant public health problem, above all that wounds fail to heal properly and become chronic. Due to its reepithelization action, insulin has the potential to heal skin lesions, by stimulating the proliferation and migration of keratinocytes, angiogenic stimulus, and increasing collagen deposition. In the present study insulin was complexed with 2-hydroxypropyl-β-cyclodextrin (HPβCD) and its wound healing effect and inclusion complex (HPβCD-I) were evaluated in excisional wounds in the skin of rats. Material and methods Three different gel based pharmaceutical forms were created: carbopol 940® base gel, an insulin gel comprising the base gel plus 50 IU of insulin and a gel complex comprising the base gel plus (HPβCD) complexed with insulin (HPβCD-I) were used to verify wound healing in vitro and in vivo assays. Results The wounds in the skin of rats were treated with gel containing HPβCD-I not cytoxically irritating and cytotoxic. Analysis of cell proliferation and measurement of the length and thickness of the epidermis showed that HPβCD-I prolonged the proliferation and migration of keratinocytes. Revascularization analysis of lesions treated with HPβCD-I compared to those treated with insulin found that angiogenic stimulus was less intense, but more constant and prolonged in the modified release process. There was increased deposition of type I and III collagen fibers in accordance with the treatment time. Conclusion Therefore, the slow release of complexed insulin modulated the reepithelialization process by stimulating cell proliferation and migration of keratinocytes, favoring greater concentration of serum insulin, modulating inflammatory response, matrix remodeling and promoting neovascularization. Angiogenesis extended by the steady release of insulin can be effective in the treatment of chronic wounds.

The effects of starter diet (days 1 to 21) supplemented with arginine (Arg) on the production performance and duodenum and jejunum mucosa morphometry of broilers were studied. Male Cobb broiler chickens (990) were randomly assigned to one of five treatments in a complete random design. Measurements of 33 chicks per treatment were made in six repetitions. The treatments consisted of a basal diet with 1.390% digestible Arg (no supplementation) and four dietary levels (1.490%, 1.590%, 1.690%, and 1.790%), providing a relationship with lysine of 1.103; 1.183; 1.262; 1.341 and 1.421%, respectively. From the age of 22 days on, all birds received conventional grower diet. The data were submitted to regression analysis by polynomial decomposition of the degrees of freedom in relation to the levels of Arg. The Arg supplementation increased (P0.05) in the growth phase (days 22 to 42) in the absence of the Arg supplementation. The supplementation of Arg over of NRC recommendation during the starter phase may be necessary for the expression of the maximal weight gain potential in birds. No effect (P0,05) na fase de crescimento (22 a 42 dias) na ausência de suplementação de Arg. A suplementação acima do recomendado pelo NRC na fase inicial pode ser necessária para a expressão máxima do potencial de ganho de peso em aves. Não houve efeito da suplementação de Arg na dieta no peso e comprimento do intestino delgado em nenhuma idade. No entanto, a relação vilo:cripta no duodeno aumentou e a profundidade da cripta diminuiu na primeira semana em resposta ao incremento de Arg na dieta. Concluiu-se que em frangos de corte a suplementação de Arg na dieta inicial melhorou o desempenho e a morfometria do intestino delgado, especialmente na primeira semana.

The genus Fusarium is known to produce mycotoxins that cause fusariosis in plants, animals and humans. Mycotoxins are among the virulence factors of this genus. Metabolic extracts of Fusarium oxysporum, isolated from a patient with onychomycosis and sterilized by filtration or autoclave, were inoculated intradermally into Wistar rats at concentrations of 0.25, 0.5 and 1 µg/µL, and the effects on their tegument were observed at 24 and 72 hours. After histological procedures and staining by hematoxylineosin, the sections were studied for their inflammatory-reaction intensity and for evidence of injury and tissue distortion. Inflammatory reactions in the dermis and the subcutaneous tissue were observed at all concentrations of the inoculated extract tested. There was a significant influx of neutrophils, mastocytes and lymphocytes, as well as a large quantity of macrophages. Apoptotic bodies and hyperemic blood vessels were observed. This reaction was directly related to the extract concentration, and was most intense in animals that received the 1 mg/µL dose. The maximum peak was observed at 24 hours. The autoclaved metabolic extract produced the same effects as the untreated one, indicating the presence of heat-resistant metabolites. In conclusion, the metabolic extracts obtained from sterilized culture filtrates of F. oxysporum are capable of inducing an inflammatory response within 24 hours in the dermis and subcutaneous tissue of rats.

The aim of this study was to evaluate morphologically the effect of teriparatide on induced orthodontic movement of the maxillary first molars in ovariectomized rats.Ovariectomized Wistar rats (n = 16), ovariectomized rats treated with teriparatide (n = 16), and nonovariectomized rats (n = 16) had orthodontic tooth movement for 5 and 7 days. The group treated with teriparatide received a subcutaneous injection (Forteo, Eli Lilly, Indianapolis, Ind; 30 μg/kg/day) for 90 days after the ovariectomy. Histologic sections obtained from the maxilla were prepared for the morphometric analysis of dental movement, the thickness of the periodontal ligament, and the number of osteoclasts in the pressure and tension areas of the apex of the root and alveolar crest in the distal root of the maxillary first molars.The ovariectomized rats treated with teriparatide had similar responses at 5 and 7 days after the induced dental movements compared with the untreated ovariectomized group. Both ovariectomized groups had greater molar movement on day 7 day compared with the controls (P 0.05). There were no statistically significant differences between groups in the spacing of the periodontal ligament or the number of osteoclasts in the areas studied.These data suggest that the treatment of osteoporosis with teriparatide is a good alternative for patients undergoing orthodontic treatment.

All methods to detect experimental loss of bone present technique limitations. The sensitivities of image and histological analyses to detect the effects of teriparatide in rats with bone loss after ovariectomy were evaluated. All methods were qualitatively valid. The standardization of methods to assess bone loss after ovariectomy is crucial to establish the degree of experimental osteoporosis. In general, methods per image or histological techniques are used. To validate these two ways to determine the degree of bone loss in ovariectomized rats, we evaluated the sensitivities of bone densitometry, conventional radiography, and histological analysis of the area occupied by collagen, detecting the effects of teriparatide treatment in the femur of ovariectomized rats with bone loss. Wistar rats were divided into three groups: a control group, in which the animals were only subjected to laparotomy; an ovariectomized group, in which bilateral removal of the ovaries was performed; and an ovariectomized + teriparatide group, in which bilateral removal of the ovaries was performed, and the animals were treated with 3 μg/100 g/day of teriparatide. Three months following the ovariectomy, bone densitometry, radiographic densitometry, and histological analysis of the area occupied by collagen fibers were carried out in the femur diaphysis. The bone densitometry revealed 11.2% reduction in femur density; in the conventional radiography, the loss of bone mass was 14.5%, and with the histological analysis, a 40.9% reduction in the area occupied by collagen was detected in the femur diaphysis. In conclusion, histological analysis could not be quantitatively compared with the methods of bone densitometry and conventional radiography; however, all of these methods were qualitatively valid for assessing the loss of bone stemming from ovariectomy and the therapeutic effect of teriparatide in the prevention of osteoporosis.