RESUMO
Chronic wounds pose significant health concerns. Current treatment options include natural compounds like natural rubber latex (NRL) from Hevea brasiliensis. NRL, particularly the F1 protein fraction, has demonstrated bioactivity, biocompatibility, and angiogenic effects. So far, there is no study comparing F1 protein with total NRL serum, and the necessity of downstream processing remains unknown. Here, we evaluated the angiogenic potential of F1 protein compared to total NRL serum and the need for downstream processing. For that, ion exchange chromatography (DEAE-Sepharose), antioxidant activity, physicochemical characterization, cell culture in McCoy fibroblasts, and wound healing in Balb-C mice were performed. Also, the evaluation of histology and collagen content and the levels of inflammatory mediators were quantified. McCoy fibroblast cell assay showed that F1 protein (0.01 %) and total NRL serum (0.01 %) significantly increased cell proliferation by 47.1 ± 11.3 % and 25.5 ± 2.5 %, respectively. However, the AA of F1 protein (78.9 ± 0.8 %) did not show a significant difference compared to NRL serum (77.0 ± 1.1 %). F1 protein and NRL serum were more effective in wound management in rodents. Histopathological analysis confirmed accelerated healing and advanced tissue repair. Similarly, the F1 protein (0.01 %) increased collagen, showing that this fraction can stimulate the synthesis of collagen by fibroblastic cells. Regarding cytokines production (IL-10, TNF-α, IFN-γ), F1 protein and NRL serum did not exert an impact on the synthesis of these cytokines. Furthermore, we did not observe statistically significant changes in dosages of enzymes (MPO and EPO) among the groups. Nevertheless, Nitric Oxide dosage was reduced drastically when the F1 protein (0.01 %) protein was applied topically. These findings contribute to the understanding of F1 protein and NRL serum properties and provide insights into cost-effectiveness and practical applications in medicine and biotechnology. Therefore, further research is needed to assess the economic feasibility of downstream processing for NRL-based herbal medicine derived from Hevea brasiliensis.
Assuntos
Hevea , Borracha , Animais , Camundongos , Látex , Hevea/química , Cicatrização , Colágeno , CitocinasRESUMO
Immune system hyperactivation is involved with clinical severity and pathological alterations during scorpion envenomation. In a murine model, mice inoculated with a lethal dose of Tityus serrulatus scorpion venom presented mitochondrial swelling in cardiomyocytes, with other structures such as sarcomeres and intercalated disks preserved. Treatment with dexamethasone or knockout animals to the interleukin-1ß receptor do not undergo mitochondrial changes in cardiomyocytes during envenomation.
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Picadas de Escorpião , Venenos de Escorpião , Animais , Camundongos , Miócitos Cardíacos , Dilatação Mitocondrial , Modelos Animais de Doenças , Venenos de Escorpião/toxicidade , EscorpiõesRESUMO
The purpose of this study is to evaluate the effects of radiofrequency (RF) on sagging skin. This is a case series study with five volunteers who received a single application of capacitive RF (BTL-6000 TR-Therapy Pro®) in the right infraumbilical abdominal region, with epidermal temperature above 40°C, for 10 min (2 min per applicator area), and the skin of the contralateral region was used as control. After 30 days, on average, the skin of the abdominal region was collected for histological analysis and stained with hematoxylin and eosin, Picro-sirus, and Verhoff. The percentage of collagen and elastic fibers found was marked by the Image J®. The statistical analysis was performed in the SPSS program (version 20), with a significance level of 95%. This was registered with the ethics and research comitee of UFTM n 3.461.688 on Jul 12, 2019 and clinical trial registration n. NCT04182542, retrospectively registered. Morphometric analysis demonstrated a remodeling of collagen and elastic fibers on the side treated with RF; however, the morphometry for collagen showed no significant difference, with an average percentage of 60.94 ± 0.32 for the control side and 61.97 ± 2.80 for the treated with p=0.32. Similarly, elastic fibers also showed no significant difference between groups, with a mean percentage of 5.67 ± 2.70 for control and 6.21 ± 2.01 for treated with p=0.19. The RF with the parameters used in this study was able to cause morphological changes in collagen and elastic fibers of the abdominal region skin; however, it showed no change in the percentage of these fibers.
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Tecido Elástico , Pele , Colágeno , Derme , Humanos , Ondas de RádioRESUMO
BACKGROUND: Chronic infection by HIV evolves with a vascular inflammatory action causing endothelial dysfunction. The action of the virus, as well as the side effects of antiretroviral drugs, contribute to the progression of cardiovascular diseases. The present study aimed to evaluate the percentage of collagen fibers and the density of mast cells, chymase and tryptase, in aortas of patients with and without HIV, and also patients with and without atherosclerosis. METHODS: Aortic fragments were obtained from autopsied patients aged 22-69 years and selected regardless of the cause of death or underlying disease. The samples were divided into four groups, (1) Group with HIV and with atherosclerosis; (2) Group with HIV and without atherosclerosis; (3) Group without HIV and with atherosclerosis; (4) Group without HIV and without atherosclerosis (Control). The percentage of collagen fibers was analyzed in the intima-media layer and the density of mast cells was analyzed in all aortic layers. Graphpad Prism 5.0® software was used for statistical analysis. RESULTS: There were more collagen fibers in HIV patients, with or without atherosclerosis. The group with HIV and atherosclerosis presented a higher density of chymase and tryptase mast cells. The correlation between collagen fibers and age was negative in the non-HIV group and with atherosclerosis. CONCLUSION: The inflammatory process resulting from HIV infection may be relevant in the alteration of aortic collagen fibers and in triggering or accelerating atherosclerosis. The study is important because HIV patients have increased risks for the development of cardiovascular diseases, and follow-up is necessary to prevent such diseases.
Assuntos
Aorta/anatomia & histologia , Aorta/patologia , Aterosclerose/etiologia , Aterosclerose/patologia , Colágenos Fibrilares/análise , Infecções por HIV/complicações , Infecções por HIV/patologia , Mastócitos/patologia , Adulto , Idoso , Autopsia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Podocyte injury in focal segmental glomerulosclerosis (FSGS) and minimal change disease (MCD) results from the imbalance between adaptive responses that maintain homeostasis and cellular dysfunction that can culminate in cell death. Therefore, an in situ analysis was performed to detect morphological changes related to cell death and autophagy in renal biopsies from adult patients with podocytopathies. Forty-nine renal biopsies from patients with FSGS (n = 22) and MCD (n = 27) were selected. In situ expression of Wilms Tumor 1 protein (WT1), light chain microtubule 1-associated protein (LC3) and caspase-3 protein were evaluated by immunohistochemistry. The foot process effacement and morphological alterations related to podocyte cell death and autophagy were analyzed with transmission electronic microscopy. Reduction in the density of WT1-labeled podocytes was observed for FSGS and MCD cases as compared to controls. Foot process width (FPW) in control group was lower than in cases of podocytopathies. In FSGS group, FPW was significantly higher than in MCD group and correlated with proteinuria. A density of LC3-labeled podocytes and the number of autophagosomes in podocytes/ pedicels were higher in the MCD group than in the FSGS group. The number of autophagosomes correlated positively with the estimated glomerular filtration rate in cases of MCD. The density of caspase-3-labeled podocytes in FSGS and MCD was higher than control group, and a higher number of podocytes with an evidence of necrosis was detected in FSGS cases than in MCD and control cases. Podocytes from patients diagnosed with FSGS showed more morphological and functional alterations resulting from a larger number of lesions and reduced cell adaptation.
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Glomerulosclerose Segmentar e Focal/patologia , Nefrose Lipoide/patologia , Podócitos/patologia , Adulto , Autofagossomos/metabolismo , Autofagia , Estudos de Casos e Controles , Caspase 3/metabolismo , Feminino , Taxa de Filtração Glomerular , Glomerulosclerose Segmentar e Focal/metabolismo , Humanos , Rim/metabolismo , Rim/patologia , Masculino , Proteínas Associadas aos Microtúbulos/metabolismo , Pessoa de Meia-Idade , Necrose , Nefrose Lipoide/metabolismo , Podócitos/citologia , Podócitos/metabolismo , Proteinúria/complicações , Proteínas WT1/metabolismoRESUMO
BACKGROUND: Carboxytherapy is capable of inducing tissue repair which results in an increase in elastic and collagen fibers. OBJECTIVES: To evaluate the effects of carboxytherapy upon human skin collagen and elastic fiber synthesis. METHODS: Case study of nine volunteers who received a single application of carboxytherapy in the left infraumbilical region, with infusion rate of 100 mL/min and 0.6 mL/kg weight over an area of 25 cm2 . After 60 days on average, the skin was collected for histological analysis and stained with picrosirius red for collagen and Verhoeff for elastic fibers. The percentage of fibers found was marked by the Image J® program and recorded in a Microsoft Excel spreadsheet. Statistical analysis was performed using SPSS (version 20), with a significance level of 95%. RESULTS: An increase in the collagen and elastic fibers sample was observed in the treated group. Morphometrically, a significant increase in the percentage of collagen in the Carboxytherapy group (41.44 ± 4.50%) was observed compared with the control group (37.44 ± 3.87%) with P = .04; for elastic fibers, the percentage showed no significant difference between the control group (10.55 ± 4.33%) and the carboxytherapy group (10.44 ± 3.71%). CONCLUSIONS: Carboxytherapy with the parameters used in this study was able to stimulate collagen and elastic fiber synthesis, with significant differences in the morphometry for collagen fibers.
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Dióxido de Carbono , Pele , Colágeno , Tecido Elástico , Humanos , CicatrizaçãoRESUMO
Diabetes mellitus (DM) may lead to gastrointestinal motility disorders. Rodent models of DM indicate the presence of morpho-functional abnormalities of the enteric nervous system. Here, we evaluated whether experimental DM can cause changes in the excitatory cholinergic fibers, neuronal density, and voltage-gated sodium channel (Nav) expression in the myenteric plexus of the ileum. After streptozotocin-induced hyperglycemia in female rats progressed for eight weeks, triple immunofluorescence labeling experiments revealed that the neuronal density in DM rats was significantly lower than that in control. On average, the density of total neurons reduced by 52.2% (pâ¯=â¯0.0001), cholinergic neurons by 50.0% (pâ¯=â¯0.0068), and nitrergic neurons by 54.8% (pâ¯=â¯0.0042). The number of neurons per ganglionic area was also significantly reduced (to 28.2% of total neurons, pâ¯=â¯0.0002; 27.7% of cholinergic neurons, pâ¯=â¯0.0002, and 32.1% of nitrergic neurons, pâ¯=â¯0.0016). Furthermore, the density of the cholinergic fibers at the surface of the longitudinal muscle was significantly reduced (DM: 24⯱â¯3%; pâ¯=â¯0.003, control: 41⯱â¯2%); however, western-blot analysis did not indicate a reduction in the expression of choline acetyltransferase (ChAT) in the DM group. The Nav1.6 isoform was detected in different myenteric neurons of the ileum. RT-qPCR data did not suggest an alteration of transcripts for ChAT, neuronal nitric oxide synthase, Nav1.3, Nav1.6, or Nav1.7. Our data support the view that chronic DM leads to a reduction of excitatory cholinergic fibers and neuronal density. However, changes in sodium channel expression pattern, which could cause neuronal dysfunction, were not detected.
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Diabetes Mellitus Experimental/metabolismo , Sistema Nervoso Entérico/metabolismo , Plexo Mientérico/fisiologia , Animais , Colina O-Acetiltransferase/metabolismo , Neurônios Colinérgicos/metabolismo , Modelos Animais de Doenças , Sistema Nervoso Entérico/fisiologia , Feminino , Regulação da Expressão Gênica/genética , Íleo/inervação , Íleo/metabolismo , Plexo Mientérico/metabolismo , Neurônios Nitrérgicos/metabolismo , Óxido Nítrico Sintase Tipo I/metabolismo , Ratos , Ratos Wistar , Canais de Sódio/genética , Canais de Sódio/metabolismo , Estreptozocina/farmacologiaRESUMO
Objective. To evaluate the expression of inflammatory markers in experimental renal failure after fetal programming. Methods. The offspring aged two and five months were divided into four groups: CC (control dams, control offspring); DC (diabetic dams, control offspring); CFA (control dams, folic acid offspring, 250 mg/Kg); and DFA (diabetic dams, folic acid offspring). Gene expression of inflammatory markers MCP-1, IL-1, NOS3, TGF-ß, TNF-α, and VEGF was evaluated by RT-PCR. Results. MCP-1 was increased in the CFA and DFA groups at two and five months of age, as well as in DC5 when compared to CC5. There was a higher expression of IL-1 in the CFA2, DFA2, and DC2 groups. There was a decrease in NOS3 and an increase in TNF-α in DFA5 in relation to CFA5. The gene expression of TGF-ß increased in cases that had received folic acid at two and five months, and VEGF decreased in the CFA5 and DFA5 groups. DC5 showed increased VEGF expression in comparison with CC5. Conclusions. Gestational diabetes mellitus and folic acid both change the expression of inflammatory markers, thus demonstrating that the exposure to harmful agents in adulthood has a more severe impact in cases which underwent fetal reprogramming.
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Diabetes Mellitus Experimental/patologia , Diabetes Gestacional/patologia , Desenvolvimento Fetal/fisiologia , Ácido Fólico/farmacologia , Rim/patologia , Insuficiência Renal/patologia , Animais , Biomarcadores/metabolismo , Quimiocina CCL2/metabolismo , Feminino , Interleucina-1/metabolismo , Rim/imunologia , Linfotoxina-alfa/metabolismo , Masculino , Óxido Nítrico Sintase Tipo III/metabolismo , Gravidez , Ratos , Ratos Wistar , Insuficiência Renal/imunologia , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Dermatophytes are fungal pathogens that cause cutaneous infections such as onychomycosis and athlete's foot in both healthy and immunocompromised patients.Trichophyton rubrum is the most prevalent dermatophyte causing human nail and skin infections worldwide, and because of its anthropophilic nature, animal infection models are limited. The purpose of this work was to compare the expression profile of T. rubrum genes encoding putative virulence factors during growth in ex vivo and in vitro infection models. The efficiency of the ex vivo skin infection model was confirmed by scanning electron microscopy (SEM), which showed that the conidia had produced hyphae that penetrated into the epidermis. Quantitative RT-PCR (qRT-PCR) analysis showed that the expression of some genes is modulated in response to the infection model used, as compared to that observed in cells grown in glucose-containing media. We concluded that ex vivo infection models help assess the molecular aspects of the interaction of T. rubrum with the host milieu, and thus provide insights into the modulation of genes during infection.
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Interações Hospedeiro-Patógeno , Modelos Biológicos , Onicomicose/microbiologia , Trichophyton , Perfilação da Expressão Gênica , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/fisiologia , Humanos , Unhas/microbiologia , Trichophyton/genética , Trichophyton/patogenicidade , Fatores de VirulênciaRESUMO
Physical activity evokes well-known adaptations in the cardiovascular system. Although exercise training induces cardiac remodeling, whether multipotent stem cells play a functional role in the hypertrophic process remains unknown. To evaluate this possibility, C57BL/6 mice were subjected to swimming training aimed at achieving cardiac hypertrophy, which was morphologically and electrocardiographically characterized. Subsequently, c-Kit(+)Lin(-) and Sca-1(+)Lin(-) cardiac stem cells (CSCs) were quantified using flow cytometry while cardiac muscle-derived stromal cells (CMSCs, also known as cardiac-derived mesenchymal stem cells) were assessed using in vitro colony-forming unit fibroblast assay (CFU-F). Only the number of c-Kit(+)Lin(-) cells increased in the hypertrophied heart. To investigate a possible extracardiac origin of these cells, a parabiotic eGFP transgenic/wild-type mouse model was used. The parabiotic pairs were subjected to swimming, and the wild-type heart in particular was tested for eGFP(+) stem cells. The results revealed a negligible number of extracardiac stem cells in the heart, allowing us to infer a cardiac origin for the increased amount of detected c-Kit(+) cells. In conclusion, the number of resident Sca-1(+)Lin(-) cells and CMSCs was not changed, whereas the number of c-Kit(+)Lin(-) cells was increased during physiological cardiac hypertrophy. These c-Kit(+)Lin(-) CSCs may contribute to the physiological cardiac remodeling that result from exercise training.
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Cardiomegalia/patologia , Cardiomegalia/fisiopatologia , Miocárdio/patologia , Condicionamento Físico Animal , Proteínas Proto-Oncogênicas c-kit/metabolismo , Células-Tronco/citologia , Células-Tronco/metabolismo , Animais , Cardiomegalia/diagnóstico por imagem , Adesão Celular , Contagem de Células , Ensaio de Unidades Formadoras de Colônias , Citometria de Fluxo , Proteínas de Fluorescência Verde/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Resistência Física , Células Estromais/citologia , Natação , Ultrassonografia , Remodelação VentricularRESUMO
OBJECTIVE: To evaluate the effects of folic acid (FA)-induced renal failure in young offspring of diabetic mothers. METHODS: The offspring of streptozotocin-induced diabetic dams were divided into four groups: CC (controls receiving vehicle); DC (diabetics receiving vehicle); CA (controls receiving FA solution, 250 mg/kg) and DA (diabetics receiving FA solution, 250 mg/kg). Renal function tests and morphometry results were analyzed. RESULTS: An increase in creatinine and urea levels was observed in CA and DA groups at two and five months. FA administration caused a significant reduction in the number of glomeruli in the offspring of diabetic dams. The diabetes group treated with FA had fewer glomeruli compared to controls at two and five months. FA caused an increase in the area of the urinary space both in controls and offspring of diabetic dams at two and five months. The number of glomeruli and area of the urinary space at two months were negatively correlated. CONCLUSIONS: Fetal programing promotes remarkable changes in kidney morphology and function in offspring. We suggest that the morphological changes in the kidneys are more pronounced when fetal programing is associated with newly acquired diseases, e.g. renal failure induced by FA.
Assuntos
Injúria Renal Aguda/embriologia , Injúria Renal Aguda/patologia , Diabetes Mellitus Experimental/patologia , Desenvolvimento Fetal , Gravidez em Diabéticas/patologia , Efeitos Tardios da Exposição Pré-Natal/patologia , Injúria Renal Aguda/fisiopatologia , Animais , Pressão Sanguínea , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/embriologia , Diabetes Mellitus Experimental/fisiopatologia , Feminino , Desenvolvimento Fetal/efeitos dos fármacos , Frequência Cardíaca , Rim/fisiopatologia , Testes de Função Renal , Gravidez , Gravidez em Diabéticas/fisiopatologia , Efeitos Tardios da Exposição Pré-Natal/fisiopatologia , Ratos , Ratos Wistar , EstreptozocinaRESUMO
Confocal microscopy provides a unique modality to examine the expression and localization of biomolecules in a variety of settings. Using this technique, an image is acquired from the focal plane of the objective using focused laser light, making it possible to work within the resolution limit of the optical system. In addition, by acquiring multiple images from a variety of focal planes, stacked series of images can provide clear spatial localization of a probed structure or protein. We describe herein the immunofluorescence methods for galectin staining in frozen sections of tissue for galectin localization using confocal microscopy.
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Galectinas/metabolismo , Microscopia Confocal/métodos , Imunofluorescência , Galectinas/análise , Vidro/química , Transporte ProteicoRESUMO
This study evaluated, by SEM, the morphology of human primary teeth roots. Twenty-four teeth were divided into 3 groups: pulp vitality (group I) and pulp necrosis without (group II) and with apical periodontitis (group III). Roots were analyzed by the presence of periodontal ligament (PDL) fibers and resorption areas. In groups I and II, presence of PDL fibers and absence of resorption were observed in all cases (100%), while all specimens (100%) of group III showed no PDL fibers and resorption areas. In conclusion, there are morphological differences in the apical region of primary teeth with different pulpal and periapical pathologies.
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Polpa Dentária/ultraestrutura , Microscopia Eletrônica de Varredura/métodos , Raiz Dentária/ultraestrutura , Dente Decíduo/ultraestrutura , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Necrose , Ligamento Periodontal/ultraestruturaRESUMO
Os hemangiomas säo tumores benignos de origem vascular, que somam 7 por cento dos tumores benignos, sendo que 56 por cento ocorrem na regiäo da cabeça e pescoço. É apresentado um caso clínico no lábio inferior com diagnóstico histopatológico de hemangioma misto. Säo discutidas as diferentes formas de diagnóstico clínico e tratamento para esse tumor, seus diferentes aspectos histopatológicos; assim como os cuidados que o Cirurgiäo-Dentista deve ter quando diante de tal lesäo
Assuntos
Humanos , Masculino , Feminino , Pessoa de Meia-Idade , HemangiomaRESUMO
Os autores descrevem a seqüência do diagnóstico e tratamento de um caso clínico de infecçäo purulenta de origem dentária com acometimento extrabucal, cuja etiologia näo foi determinada corretamente por outros profissionais que inicialmente atenderam o paciente, os quais instituiram tratamento incorreto que resultou em fracasso. O diagnóstico e tratamento empregados säo discutidos, assim como as possíveis complicaçöes que esta entidade clínica pode ocasionar