RESUMEN
This study aimed to develop bone regenerative therapeutic strategies, based on the addition of bone marrow stromal cells (BMSC) on bioglass/collagen (BG/COL) scaffolds. For this purpose, an in vivo study was conducted using tissue response of the BG/COL scaffolds combined with BMSC in a critical-size defects. Wistar rats were submitted to the surgical procedure to perform the cranial critical size bone defects and distributed in four groups (20 animals per group): Control Group (CG) (rats submitted to the cranial bone defect surgery without treatment), Bioglass Group (BG) (rats treated with BG), BG/COL Group (rats treated with BG/COL) and Bioglass/Collagen and BMSC Group (BG/COL/BMSC) (rats treated with BG/COL scaffolds enriched with BMSCs). Animals were euthanized 15 and 30 days after surgery. Scanning electron microscopy, histopathological and immunohistochemistry analysis were used. SEM analysis demonstrated that porous scaffolds were obtained, and Col fibers were successfully impregnated to BG matrices. The implantation of the BMSC on BG/COL based scaffolds was effective in stimulating newly bone formation and produced an increased immunoexpression of markers related to the bone repair. These results highlight the potential of BG/COL scaffolds and BMSCs to be used as a therapeutic approach for bone regeneration.
Asunto(s)
Células Madre Mesenquimatosas , Andamios del Tejido , Ratas , Animales , Ratas Wistar , Colágeno/farmacología , Osteogénesis , Regeneración Ósea , Modelos Teóricos , Células de la Médula Ósea , Ingeniería de Tejidos/métodosRESUMEN
Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (<3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 µg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, α-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.
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Phaseolus , Animales , Ratas , Peso Molecular , Óxido Nítrico , Ratas Wistar , Péptidos , TirosinaRESUMEN
Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (<3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 µg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, α-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.
RESUMEN
About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction <3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 µg/mL PV3. Oxidative stress was provoked by 3% H2O2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO- and endothelium-dependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 µg/mL PV3. PV3 at 30 µg/mL increased cell viability, while cytotoxicity was observed only with 250 µg/mL PV3. PV3 at 10 µg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H2O2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.
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Phaseolus , Antioxidantes/farmacología , Endotelio , Humanos , Peróxido de Hidrógeno , Peso Molecular , Péptidos/farmacologíaRESUMEN
Nucleotides are important to cell growth and division and are crucial to the rapid proliferation of such cells as the intestinal mucosa and immune cells. Accordingly, the nucleotide requirements of animals are high during periods of rapid growth and periods of stress like post-weaning period. Thus, nucleotide supplementation may be a possible alternative to in-feed antibiotics as growth promoter in this phase. The study aimed to evaluate dietary nucleotide supplementation as an alternative to in-feed antibiotics on performance and gut health of weaned piglets. Ninety-six 21-day-old piglets, weighing 7.44⯱â¯0.65â¯kg, were allocated into 1 of 3 treatments (8 pens per treatment; 4 pigs per pen) in a 14-day trial. Dietary treatments consisted of control: corn-soybean meal-based diet; nucleotides: control +2â¯g/kg of a nutritional additive with purified nucleotides; and antibiotic: control +0.8â¯g/kg of antibiotic growth promoter based on colistin and tylosin. Performance variables and fecal score were not affected (Pâ¯>â¯0.05) by supplementing nucleotide or antibiotic. Nucleotides treatment had similar effect to antibiotic and superior to control (Pâ¯<â¯0.05) on enhancing duodenum villus height, jejunum crypt depth, and reduction of Paneth cellular area. Duodenum and ileum of animals supplemented with nucleotides or antibiotics had higher (Pâ¯<â¯0.05) number of proliferating cells than did those of control animals, whereas the jejunum of animals that received antibiotic diets presented more (Pâ¯<â¯0.05) proliferating cells than either the nucleotides or control animals. Jejunum of nucleotide-treated piglets showed a greater number of apoptotic cells than those fed antibiotic or control diets (Pâ¯<â¯0.05). Nucleotides and antibiotic treatments decreased the B lymphocyte counts in duodenum and ileum (Pâ¯<â¯0.05) but increased in the jejunum (Pâ¯<â¯0.05), when compared to the control treatment. Relative abundance of mitogen-activated protein kinases-6, haptoglobin, and tumor necrosis factor-α mRNA was not influenced (Pâ¯>â¯0.05) by treatments. In the ileal, antibiotic supplementation reduced total bacteria quantification compared to nucleotide supplementation or the control (Pâ¯<â¯0.05), whereas nucleotides supplementation increased enterobacteria proliferation compared to the antibiotic or control diets (Pâ¯<â¯0.05). However, nucleotides and antibiotic reduced (Pâ¯<â¯0.05) colon total bacteria quantification when compared to control. These results suggest that the nucleotides source used to weaned piglets improved gut health by modulating the local immune response and modulating intestinal mucosa development, and, therefore, nucleotides may be an alternative to antibiotics as growth promoters.
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Alimentación Animal , Antibacterianos , Alimentación Animal/análisis , Animales , Dieta/veterinaria , Suplementos Dietéticos , Mucosa Intestinal , Nucleótidos , Porcinos , DesteteRESUMEN
Compensatory hypertrophy (CH) occurs due to excessive mechanical load on a muscle, promoting an increase in the size of muscle fibers. In clinical practice, situations such as partial nerve injuries, denervation, and muscle imbalance caused by trauma to muscles and nerves or diseases that promote the loss of nerve conduction can induce CH in muscle fibers. Photobiomodulation (PBM) has demonstrated beneficial effects on muscle tissue during CH. The aim of the present study was to evaluate the effect of PBM on the inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) as well as type 2 metalloproteinases (MMP-2) during the process of CH due to excessive load on the plantaris muscle in rats. Forty-five Wistar rats weighing 250 g were divided into three groups: control group (n = 10), hypertrophy (H) group (n = 40), and H + PBM group (n = 40). CH was induced through the ablation of synergist muscles of the plantaris muscle. The tendons of the gastrocnemius and soleus muscles were isolated and sectioned to enable the partial removal of each of muscle. The preserved plantaris muscle below the removed muscles was submitted to excessive functional load. PBM was performed with low-level laser (AsGaAl, λ = 780 nm; 40 mW; energy density: 10 J/cm2; 10 s on each point, 8 points; 3.2 J). Animals from each group were euthanized after 7 and 14 days. The plantaris muscles were carefully removed and sent for analysis of the gene and protein expression of IL-6 and TNF-α using qPCR and ELISA, respectively. MMP-2 activity was analyzed using zymography. The results were submitted to statistical analysis (ANOVA + Tukey's test, p < 0.05). The protein expression analysis revealed an increase in IL-6 levels in the H + PBM group compared to the H group and a reduction in the H group compared to the control group. A reduction in TNF-α was found in the H and H + PBM groups compared to the control group at 7 days. The gene expression analysis revealed an increase in IL-6 in the H + PBM group compared to the H group at 14 days as well as an increase in TNF-α in the H + PBM group compared to the H group at 7 days. Increases in MMP-2 were found in the H and H + PBM groups compared to the control group at both 7 and 14 days. Based on findings in the present study, it is concluded that PBM was able to modulate pro-inflammatory cytokines that are essential for the compensatory hypertrophy process. However, it has not shown a modulation effect directly in MMP-2 activity during the same period evaluated.
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Citocinas/metabolismo , Regulación de la Expresión Génica/efectos de la radiación , Terapia por Luz de Baja Intensidad , Músculo Esquelético/patología , Músculo Esquelético/efectos de la radiación , Animales , Hipertrofia/metabolismo , Hipertrofia/patología , Hipertrofia/radioterapia , Interleucina-6/metabolismo , Masculino , Metaloproteinasa 2 de la Matriz/metabolismo , Músculo Esquelético/metabolismo , Ratas , Ratas Wistar , Tendones/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction <3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 µg/mL PV3. Oxidative stress was provoked by 3% H2O2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO- and endothelium-dependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 μg/mL PV3. PV3 at 30 μg/mL increased cell viability, while cytotoxicity was observed only with 250 μg/mL PV3. PV3 at 10 μg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H2O2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.
Asunto(s)
Humanos , Phaseolus , Péptidos/farmacología , Endotelio , Peróxido de Hidrógeno , Peso Molecular , Antioxidantes/farmacologíaRESUMEN
INTRODUCTION: Collagen from marine esponges has been used as a promising material for tissue engineering proposals. Similarly, photobiomodulation (PBM) is able of modulating inflammatory processes after an injury, accelerating soft and hard tissue healing and stimulating neoangiogenesis. However, the effects of the associated treatments on bone tissue healing have not been studied yet. In this context, the present study aimed to evaluate the biological temporal modifications (using two experimental periods) of marine sponge collagen or sponging (SPG) based scaffold and PBM on newly formed bone using a calvaria bone defect model. MATERIAL AND METHODS: Wistar rats were distributed into two groups: SPG or SPG/PBM and euthanized into two different experimental periods (15 and 45 days post-surgery). A cranial critical bone defect was used to evaluate the effects of the treatments. Histology, histomorfometry and immunohistological analysis were performed. RESULTS: Histological findings demonstrated that SPG/PBM-treated animals, 45 days post-surgery, demonstrated a higher amount of connective and newly formed bone tissue at the region of the defect compared to CG. Notwithstanding, no difference among groups were observed in the histomorphometry. Interestingly, for both anti-transforming growth factor-beta (TGF-ß) and anti-vascular endothelial growth factor (VEGF) immunostaining, higher values for SPG/PBM, at 45 days post-surgery could be observed. CONCLUSION: It can be concluded that the associated treatment can be considered as a promising therapeutical intervention.
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Organismos Acuáticos/química , Colágeno/farmacología , Terapia por Luz de Baja Intensidad , Cráneo/patología , Andamios del Tejido/química , Cicatrización de Heridas/efectos de los fármacos , Animales , Modelos Animales de Enfermedad , Masculino , Ratas Wistar , Cráneo/efectos de los fármacos , Factor de Crecimiento Transformador beta/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Background: Taking into account the probable role that race/skin color may have for determining outcomes in maternal health, the objective of this study was to assess whether maternal race/skin color is a predictor of severe maternal morbidity. Methods: This is a secondary analysis of the Brazilian Network for Surveillance of Severe Maternal Morbidity, a national multicenter cross-sectional study of 27 Brazilian referral maternity hospitals. A prospective surveillance was performed to identify cases of maternal death (MD), maternal near miss (MNM) events, and potentially life-threatening conditions (PLTC), according to standard WHO definition and criteria. Among 9,555 women with severe maternal morbidity, data on race/skin color was available for 7,139 women, who were further divided into two groups: 4,108 nonwhite women (2,253 black and 1,855 from other races/skin color) and 3,031 white women. Indicators of severe maternal morbidity according to WHO definition are shown by skin color group. Adjusted Prevalence Ratios (PRadj - 95%CI) for Severe Maternal Outcome (SMO=MNM+MD) were estimated according to sociodemographic/obstetric characteristics, pregnancy outcomes, and perinatal results considering race. Results: Among 7,139 women with severe maternal morbidity evaluated, 90.5% were classified as PLTC, 8.5% as MNM, and 1.6% as MD. There was a significantly higher prevalence of MNM and MD among white women. MNMR (maternal near miss ratio) was 9.37 per thousand live births (LB). SMOR (severe maternal outcome ratio) was 11.08 per 1000 LB, and MMR (maternal mortality ratio) was 170.4 per 100,000 LB. Maternal mortality to maternal near miss ratio was 1 to 5.2, irrespective of maternal skin color. Hypertension, the main cause of maternal complications, affected mostly nonwhite women. Hemorrhage, the second more common cause of maternal complication, predominated among white women. Nonwhite skin color was associated with a reduced risk of SMO in multivariate analysis. Conclusion: Nonwhite skin color was associated with a lower risk for severe maternal outcomes. This result could be due to confounding factors linked to a high rate of Brazilian miscegenation.
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Mortalidad Materna , Complicaciones del Embarazo/epidemiología , Pigmentación de la Piel , Adolescente , Adulto , Brasil/epidemiología , Femenino , Humanos , Nacimiento Vivo/epidemiología , Embarazo , Complicaciones del Embarazo/mortalidad , Resultado del Embarazo/epidemiología , Población Blanca , Adulto JovenRESUMEN
The combination of different biomaterials can be a promising intervention for the composites manufacture, mainly by adding functional and structural characteristics of each material and guarantee the advantages of the use of these composites. In this context, the aim of this study was to develop and evaluated the influence of the incorporation of marine spongin (SPG) into Biosilicate® (BS) in different proportions be used during bone repair. For this purpose, it was to develop and investigate different BS/SPG formulations for physico-chemical and morphological characteristics by pH, loss mass, Fourier transform infrared spectrometer (FTIR) and scanning electron microscope (SEM) analysis. Additionally, the influence of these composites on cell viability, proliferation, and alkaline phosphatase (ALP) activity were investigated. The results revealed that the pH values of all BS groups (with or without SPG) increased over time. A significant mass loss was observed in all composites, mainly with higher SPG percentages. Additionaly, SEM micrographies demonstrated fibers of SPG into BS and material degradation over time. Moreover, FTIR spectral analysis revealed characteristic peaks of PMMA, BS, and SPG in BS/SPG composites. BS/SPG groups demonstrated a positive effect for fibroblast proliferation after 3 and 7 days of culture. Additionally, BS and BS/SPG formulations (at 10% and 20% of SPG) presented similar values of osteoblasts viability and proliferation after 7 days of culture. Furthermore, ALP activity demonstrated no significant difference between BS and BS/SPG scaffolds, at any composition. Based on the present in vitro results, it can be concluded that the incorporation of SPG into BS was possible and produced an improvement in the physical-chemical characteristics and in the biological performance of the graft especially the formulation with 80/20 and 90/10. Future research should focus on in vivo evaluations of this novel composite.
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Materiales Biocompatibles/química , Vidrio/química , Poríferos/metabolismo , Células 3T3 , Fosfatasa Alcalina/metabolismo , Animales , Sustitutos de Huesos/química , Línea Celular , Proliferación Celular , Supervivencia Celular , Concentración de Iones de Hidrógeno , Ensayo de Materiales , Ratones , Microscopía Electrónica de Rastreo , Osteoblastos/metabolismo , Espectroscopía Infrarroja por Transformada de Fourier , Ingeniería de Tejidos/métodos , Andamios del TejidoRESUMEN
Biomaterial-based bone grafts have an important role in the field of bone tissue engineering. One of the most promising classes of biomaterials is collagen, including the ones from marine biodiversity (in general, called spongin (SPG)). Also, hydroxyapatite (HA) has an important role in stimulating bone metabolism. Therefore, this work investigated the association of HA and SPG composites in order to evaluate their physico-chemical and morphological characteristics and their in vitro biological performance. For this, pre-set composite disks were evaluated by means of mass loss after incubation, pH, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and "in vitro" cell viability. pH measurements showed no statistical difference between groups. Moreover, a higher mass loss was observed for HA/SPG70/30 compared to the other groups for all experimental periods. Moreover, SEM representative micrographs showed the degradation of the samples with and without immersion. FTIR analysis demonstrated the absorption peaks for poly(methyl methacrylate) (PMMA), HA, and SPG. A higher L292 cell viability for control and PMMA was observed compared to HA and HA/SPG 90/10. Also, HA/SPG 70/30 showed higher cell viability compared to HA and HA/SPG 90/10 on days 3 and 7 days of culture. Furthermore, HA showed a significant lower MC3T3 cell viability compared to control and HA/SPG 70/30 on day 3 and no significant difference was observed between the composites in the last experimental period. Based on our investigations, it can be concluded that the mentioned composites were successfully obtained, presenting improved biological properties, especially the one mimicking the composition of bone (with 70% of HA and 30% of SPG). Consequently, these data highlight the potential of the introduction of SPG into HA to improve the performance of the graft for bone regeneration applications. Further long-term studies should be carried out to provide additional information concerning the late stages of material degradation and bone healing in the presence of HA/SPG.
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Materiales Biocompatibles/química , Sustitutos de Huesos/química , Colágeno/química , Durapatita/química , Polimetil Metacrilato/química , Andamios del Tejido , Animales , Materiales Biocompatibles/farmacología , Sustitutos de Huesos/farmacología , Huesos/citología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Colágeno/farmacología , Durapatita/farmacología , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Concentración de Iones de Hidrógeno , Ratones , Células 3T3 NIH , Polimetil Metacrilato/farmacología , Poríferos/química , Ingeniería de Tejidos/métodosRESUMEN
O objetivo do presente trabalho foi relatar um caso de leishmaniose visceral com apresentação mucosa em um cão com hiperadrenocorticismo. Um canino, macho, da raça Poodle, 11 anos de idade, foi atendido com histórico de disfagia, halitose e sialorreia. Ao exame físico, observou-se linfadenomegalia generalizada e alterações cutâneas, como rarefação pilosa, comedões, telangiectasia e atrofia cutânea. Além disso, o animal também apresentava formações orais localizadas na língua. Dos exames hematológicos e bioquímicos realizados, a única alteração encontrada foi elevação da fosfatase alcalina (1724u/L). O teste de supressão com a dexametasona em dose baixa foi executado para investigar hiperadrenocorticismo, tendo resultado positivo. Também foram realizados exames citológicos dos linfonodos, da medula óssea e das formações orais, tendo sido observada a presença de formas amastigotas de Leishmania sp. em todas as amostras. O animal foi submetido à biópsia incisional das formações orais, e a análise histopatológica demonstrou um quadro de inflamação granulomatosa com presença de grande quantidade de microrganismos morfologicamente compatíveis com formas amastigotas de Leishmania sp. no interior das células inflamatórias. Diante dos achados clínicos e dos exames complementares, diagnosticou-se um caso de leishmaniose com manifestação mucosa atípica, associado ao hiperadrenocorticismo, podendo essa endocrinopatia ter sido um fator predisponente para essa enfermidade infectocontagiosa.(AU)
The objective of the present study was to report a case of visceral leishmaniasis with mucosal presentation in a dog with hyperadrenocorticism. A canine, male, Poodle, 11 years old, was attended with a history of dysphagia, halitosis, and sialorreia. The physical examination revealed generalized lymphadenomegaly and cutaneous alterations such as hair loss, comedones, telangiectasia, and cutaneous atrophy. Futhermore, the animal also had localized oral formations on the tongue. From the hematological and biochemical tests performed, the only alteration was alkaline phosphatase elevation (1724u / L). The low dose dexamethasone suppression test was performed to investigate hyperadrenocorticism and found a positive result. In addition, cytological exams of lymph nodes, bone marrow and oral formations were also performed, and the presence of amastigote forms of Leishmania sp. were observed in all samples. The animal was submitted to incisional biopsy of the oral formations and the histopathological analysis showed a granulomatous inflammation with presence of large quantity of microorganisms morphologically compatible with amastigotes forms of Leishmania sp. within the inflammatory cells. Faced with clinical findings and complementary exams, a case of leishmaniasis with atypical mucosal manifestation, associated with hyperadrenocorticism, was diagnosed, and this endocrinopathy could have been a predisposing factor to this infectious-contagious disease.(AU)
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Animales , Perros , Hiperfunción de las Glándulas Suprarrenales/veterinaria , Perros/lesiones , Leishmaniasis Visceral/veterinaria , Terapia de InmunosupresiónRESUMEN
O objetivo do presente trabalho foi relatar um caso de leishmaniose visceral com apresentação mucosa em um cão com hiperadrenocorticismo. Um canino, macho, da raça Poodle, 11 anos de idade, foi atendido com histórico de disfagia, halitose e sialorreia. Ao exame físico, observou-se linfadenomegalia generalizada e alterações cutâneas, como rarefação pilosa, comedões, telangiectasia e atrofia cutânea. Além disso, o animal também apresentava formações orais localizadas na língua. Dos exames hematológicos e bioquímicos realizados, a única alteração encontrada foi elevação da fosfatase alcalina (1724u/L). O teste de supressão com a dexametasona em dose baixa foi executado para investigar hiperadrenocorticismo, tendo resultado positivo. Também foram realizados exames citológicos dos linfonodos, da medula óssea e das formações orais, tendo sido observada a presença de formas amastigotas de Leishmania sp. em todas as amostras. O animal foi submetido à biópsia incisional das formações orais, e a análise histopatológica demonstrou um quadro de inflamação granulomatosa com presença de grande quantidade de microrganismos morfologicamente compatíveis com formas amastigotas de Leishmania sp. no interior das células inflamatórias. Diante dos achados clínicos e dos exames complementares, diagnosticou-se um caso de leishmaniose com manifestação mucosa atípica, associado ao hiperadrenocorticismo, podendo essa endocrinopatia ter sido um fator predisponente para essa enfermidade infectocontagiosa.(AU)
The objective of the present study was to report a case of visceral leishmaniasis with mucosal presentation in a dog with hyperadrenocorticism. A canine, male, Poodle, 11 years old, was attended with a history of dysphagia, halitosis, and sialorreia. The physical examination revealed generalized lymphadenomegaly and cutaneous alterations such as hair loss, comedones, telangiectasia, and cutaneous atrophy. Futhermore, the animal also had localized oral formations on the tongue. From the hematological and biochemical tests performed, the only alteration was alkaline phosphatase elevation (1724u / L). The low dose dexamethasone suppression test was performed to investigate hyperadrenocorticism and found a positive result. In addition, cytological exams of lymph nodes, bone marrow and oral formations were also performed, and the presence of amastigote forms of Leishmania sp. were observed in all samples. The animal was submitted to incisional biopsy of the oral formations and the histopathological analysis showed a granulomatous inflammation with presence of large quantity of microorganisms morphologically compatible with amastigotes forms of Leishmania sp. within the inflammatory cells. Faced with clinical findings and complementary exams, a case of leishmaniasis with atypical mucosal manifestation, associated with hyperadrenocorticism, was diagnosed, and this endocrinopathy could have been a predisposing factor to this infectious-contagious disease.(AU)
Asunto(s)
Animales , Perros , Hiperfunción de las Glándulas Suprarrenales/veterinaria , Perros/lesiones , Leishmaniasis Visceral/veterinaria , Terapia de InmunosupresiónRESUMEN
Phototherapy has demonstrated positive effects in the treatment of peripheral nerve injury, but there is a need to investigate the dosimetric parameters. Thus, the aim of the present study was to conduct a literature review on the effects of photobiomodulation with the use of low-level laser therapy (LLLT) on the treatment of peripheral nerve injury in experimental models. The databases of PubMed/MEDLINE, SCOPUS, and SPIE Digital Library were searched for articles on the use of LLLT in experimental models of peripheral nerve injury published in English between January 2007 and March 2016. The laser parameter variability was wavelength (632.8 to 980 nm), power (10 to 190 mW), and total energy (0.15 to 90 J) in pulsed or continuous wave and single or multiple points. Eighteen original articles demonstrating the effects of LLLT on the acceleration of functional recovery, morphological aspects as well as the modulation of the expression inflammatory cytokines, and growth factors were selected. LLLT is a viable phototherapeutic modality for the treatment of peripheral nerve injury, demonstrating positive effects on the neuromuscular repair process using either red or infrared light. The majority of studies used a power of up to 50 mW and total energy of up to 15 J administered to multiple points. The determination of these parameters is important to the standardization of a LLLT protocol to enhance the regeneration process following a peripheral nerve injury.
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Terapia por Luz de Baja Intensidad/métodos , Traumatismos de los Nervios Periféricos/radioterapia , Animales , Modelos Animales de Enfermedad , Regeneración Nerviosa/efectos de la radiación , Recuperación de la FunciónRESUMEN
Bioactive glasses (BGs) are known for their ability to bond to living bone and cartilage. In general, they are readily available in powder and monolithic forms, which are not ideal for the optimal filling of bone defects with irregular shapes. In this context, the development of BG-based scaffolds containing flexible fibres is a relevant approach to improve the performance of BGs. This study is aimed at characterizing a new, highly porous, fibrous glassy scaffold and evaluating its in vitro and in vivo biocompatibility. The developed scaffolds were characterized in terms of porosity, mineralization and morphological features. Additionally, fibroblast and osteoblast cells were seeded in contact with extracts of the scaffolds to assess cell proliferation and genotoxicity after 24, 72 and 144 h. Finally, scaffolds were placed subcutaneously in rats for 15, 30 and 60 days. The scaffolds presented interconnected porous structures, and the precursor bioglass could mineralize a hydroxyapatite (HCA) layer in simulated body fluid (SBF) after only 12 h. The biomaterial elicited increased fibroblast and osteoblast cell proliferation, and no DNA damage was observed. The in vivo experiment showed degradation of the biomaterial over time, with soft tissue ingrowth into the degraded area and the presence of multinucleated giant cells around the implant. At day 60, the scaffolds were almost completely degraded and an organized granulation tissue filled the area. The results highlight the potential of this fibrous, glassy material for bone regeneration, due to its bioactive properties, non-cytotoxicity and biocompatibility. Future investigations should focus on translating these findings to orthotopic applications. Copyright © 2015 John Wiley & Sons, Ltd.
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Materiales Biocompatibles/farmacología , Vidrio/química , Ensayo de Materiales/métodos , Andamios del Tejido/química , Animales , Calcificación Fisiológica/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Concentración de Iones de Hidrógeno , Masculino , Ratones , Pruebas de Mutagenicidad , Osteoblastos/citología , Osteoblastos/efectos de los fármacos , Porosidad , Ratas Wistar , Espectroscopía Infrarroja por Transformada de Fourier , Tejido Subcutáneo/patologíaRESUMEN
The aims of this study were to characterize different BS/PLGA composites for their physicochemical and morphological characteristics and evaluate the in vitro and in vivo biological performance. The physicochemical and morphological modifications were analyzed by pH, mass loss, XRD, setting time, and SEM. For in vitro analysis, the osteoblast and fibroblast viability was evaluated. For in vivo evaluations, histopathology and immunohistochemistry were performed in a tibial defect in rats. After incubation, all composites presented lower values in pH and mass loss over time. Moreover, XRD and SEM analysis confirmed that the composites degraded over time. Additionally, pore formation was observed by SEM analysis after incubation mainly in BS/PLGA groups. BS/PLGA showed significantly increased in osteoblast viability 24 h. Moreover, BS/PLGA composites demonstrated an increase in fibroblast viability in all periods analyzed when compared to BS. In the in vivo study, after 2 and 6 weeks of implantation of biomaterials, histopathological findings revealed that the BS/PLGA composites degrades over time, mainly at periphery. Moreover, can be observed the presence of granulation tissue, bone formation, Runx-2, and RANKL immunoexpression in all groups. In conclusion, BS/PLGA composites present appropriate physicochemical characteristics, stimulate the cellular viability, and enhance the bone repair in vivo. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1063-1074, 2017.
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Ácido Láctico , Ensayo de Materiales , Osteoblastos/metabolismo , Ácido Poliglicólico , Silicatos , Tibia/metabolismo , Fracturas de la Tibia/terapia , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Fibroblastos/citología , Fibroblastos/patología , Concentración de Iones de Hidrógeno , Ácido Láctico/química , Ácido Láctico/farmacología , Ratones , Osteoblastos/patología , Osteogénesis/efectos de los fármacos , Ácido Poliglicólico/química , Ácido Poliglicólico/farmacología , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ratas , Silicatos/química , Silicatos/farmacología , Tibia/patología , Fracturas de la Tibia/metabolismo , Fracturas de la Tibia/patologíaRESUMEN
Respiratory syncytial virus (RSV)-specific CD8(+) T cell responses do not protect against reinfection. Activation of mammalian target of rapamycin (mTOR) impairs memory CD8(+) T cell differentiation. Our hypothesis was that RSV inhibits the formation of CD8(+) T cells memory responses through mTOR activation. To explore this, human and mouse T cells were used. RSV induced mTOR phosphorylation at Ser2448 in CD8 T cells. mTOR activation by RSV was completely inhibited using rapamycin. RSV-infected children presented higher mTOR gene expression on nasal washes comparing to children infected with metapneumovirus and rhinovirus. In addition, RSV-infected infants presented a higher frequency of CD8(+) pmTORser2448(+) T cells in nasal washes compared to RSV-negative infants. Rapamycin treatment increased the frequency of mouse CD8 RSV-M282-90 pentamer-positive T cells and the frequency of RSV-specific memory T cells precursors. These data demonstrate that RSV is activating mTOR directly in CD8 T cells, indicating a role for mTOR during the course of RSV infection.
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Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Líquido del Lavado Nasal/inmunología , Infecciones por Virus Sincitial Respiratorio/inmunología , Infecciones por Virus Sincitial Respiratorio/metabolismo , Virus Sincitiales Respiratorios/inmunología , Serina-Treonina Quinasas TOR/metabolismo , Animales , Linfocitos T CD8-positivos/efectos de los fármacos , Niño , Humanos , Memoria Inmunológica/efectos de los fármacos , Inmunosupresores/farmacología , Lactante , Activación de Linfocitos/efectos de los fármacos , Ratones , Líquido del Lavado Nasal/virología , Fosforilación , Infecciones por Virus Sincitial Respiratorio/virología , Sirolimus/farmacología , Serina-Treonina Quinasas TOR/genéticaRESUMEN
The mosquito Stegomyia aegypti (=Aedes aegypti) (Diptera: Culicidae) is a vector for the dengue and yellow fever viruses. As blood digestion occurs in the midgut, this organ constitutes the route of entry of many pathogens. The effects of the insecticide imidacloprid on the survival of St. aegypti were investigated and the sub-lethal effects of the insecticide on midgut development were determined. Third instar larvae were exposed to different concentrations of imidacloprid (0.15, 1.5, 3.0, 6.0 and 15.0 p.p.m.) and survival was monitored every 24 h for 10 days. Midguts from imidacloprid-treated insects at different stages of development were dissected and processed for analyses by transmission electron microscopy, immunofluorescence microscopy and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assays. Imidacloprid concentrations of 3.0 and 15.0 p.p.m. were found to affect midgut development similarly. Digestive cells of the fourth instar larvae (L4) midgut exposed to imidacloprid had more multilamellar bodies, abundantly found in the cell apex, and more electron-lucent vacuoles in the basal region compared with those from untreated insects. Moreover, imidacloprid interfered with the differentiation of regenerative cells, dramatically reducing the number of digestive and endocrine cells and leading to malformation of the midgut epithelium in adults. The data demonstrate that imidacloprid can reduce the survival of mosquitoes and thus indicate its potentially high efficacy in the control of St. aegypti populations.
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Aedes/efectos de los fármacos , Imidazoles/farmacología , Insecticidas/farmacología , Nitrocompuestos/farmacología , Aedes/crecimiento & desarrollo , Aedes/fisiología , Animales , Femenino , Larva/efectos de los fármacos , Larva/crecimiento & desarrollo , Larva/fisiología , Longevidad/efectos de los fármacos , NeonicotinoidesRESUMEN
Oxidative stress is present in severe asthma and contributes to the low response to corticoids through the downregulation of histone deacetylase (HDAC) and the increase of cytokines. Low-level laser therapy (LLLT) has been proven to be an anti-inflammatory. Thus, we investigated the laser effect on lipopolysaccharide (LPS)-induced cytokine secretion and HDAC activity in U937 cells under oxidative stress. U937 cells activated with oxidative stress were treated with dexamethasone (dexa) or laser. Cytokines and phosphoinositide 3-kinase (PI3K) were measured by ELISA whilst the HDAC was detected through colorimetric assay. LPS activated- U937 cells cytokines secretion increased with H2O2 (hydrogen peroxide) as well as with TSA (trichostatin). The HDAC activity in activated U937 cells was decreased. LLLT and dexa inhibited the LPS-stimulated U937 cells cytokines, but dexa effect disappeared with H2O2. With TSA, the LLLT was less effective on H2O2/LPS stimulated- U937 cells cytokines. Dexa failed on H2O2/LPS- induced HDAC, while LLLT restored the HDAC and the dexa effect. LLLT plus prostaglandin E2 (PGE2) increased cyclic adenosine monophosphate (cAMP) and potentiated the laser action on oxidative stress-induced cytokine. LLLT reduced the PI3K and its effects on cytokine and HDAC was suppressed with LY294002. In situations of corticoid resistance, LLLT acts decreasing the cytokines and HDAC through the activation of the protein kinase A via the inhibition of PI3K.