RESUMO
In this chapter, we report advances in tissue culture applied to Passiflora. We present reproducible protocols for somatic embryogenesis, endosperm-derived triploid production, and genetic transformation for such species knowledge generated by our research team and collaborators in the last 20 years. Our research group has pioneered the work on passion fruit somatic embryogenesis, and we directed efforts to characterize several aspects of this morphogenic pathway. Furthermore, we expanded the possibilities of understanding the molecular mechanism related to developmental phase transitions of Passiflora edulis Sims. and P. cincinnata Mast., and a transformation protocol is presented for the overexpression of microRNA156.
Assuntos
Passiflora , Técnicas de Embriogênese Somática de Plantas , Técnicas de Cultura de Tecidos , Passiflora/genética , Passiflora/crescimento & desenvolvimento , Técnicas de Embriogênese Somática de Plantas/métodos , Técnicas de Cultura de Tecidos/métodos , Transformação Genética , MicroRNAs/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Endosperma/genética , Endosperma/crescimento & desenvolvimento , Regulação da Expressão Gênica de PlantasRESUMO
The study investigated allogenic pulp transplantation as an innovative method of regenerative endodontic therapy. Three patients were selected for the endodontic treatment of single-root teeth, who also had a son/daughter with deciduous teeth or third molars scheduled for extraction. Receptor teeth were endodontically instrumented and irrigated using a tri-antibiotic solution. During the transplant procedures, the teeth from the son/daughter were extracted, sectioned, and the pulp was carefully removed. The harvested pulp from the donor was inserted into the root canal of the host tooth (father/mother), followed by direct pulp capping and resin composite restoration. The teeth were followed-up with for 2 years and were surveyed with computed tomography, the electric pulp vitality test, and Doppler ultrasound examination. At the 6-month follow-up, positive pulp vitality and the formation of periapical lesions were verified in cases 1 and 2. Case 3 showed remarkable periapical radiolucency before transplantation, but after 1 year, such lesions disappeared and there was positive vitality. All teeth were revascularized as determined by Doppler imaging after 2 years with no signs of endodontic/periodontal radiolucency. In conclusion, although this was a case series with only three patients and four teeth treated, it is possible to suppose that this allogenic pulp transplantation protocol could represent a potential strategy for pulp revitalization in specific endodontic cases.
RESUMO
Non-infectious uveitis, an ocular inflammatory condition that affects the iris, ciliary body, choroid, and adjacent tissues (retina, optic nerve, and vitreous), is an important cause of blindness worldwide. Sirolimus (SRL), a potent immunomodulatory drug, has shown promising results in the treatment of inflammatory ocular diseases. Despite this therapeutic potential, its clinical use is a major challenge due to low bioavailability and poor solubility. Poly(lactic-co-glycolic acid) (PLGA) is a biodegradable polymer commonly used for ophthalmic drug delivery due to its suitable characteristics such as biocompatibility, good mechanical properties, and improvement of the pharmacokinetic profile of the drug. In the present study, we investigated the effects of SRL-PLGA implant on experimental autoimmune uveitis in rabbits. Clinical and histopathological examinations were performed, followed by assessment of protein levels and determination of myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) activity in the aqueous humor/vitreous. As a result, treated eyes had decreased average inflammatory scores, protein significant decreases in treated eyes, assessed after 35 days. Histopathological examination showed less severe intraocular inflammation and decreased tissue damage in treated eyes. According to these results, the SRL-PLGA implant evaluated in this study was apparently safe, reducing inflammation in treated eyes, with an extended effect possibly associated with prolonged release of SRL in the posterior segment of the eye. Therefore, intravitreal SRL-PLGA implant could be a promising alternative for treatment of non-infectious uveitis.
Assuntos
Implantes de Medicamento , Imunossupressores/administração & dosagem , Sirolimo/administração & dosagem , Uveíte/tratamento farmacológico , Corpo Vítreo , Animais , Imunossupressores/uso terapêutico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Coelhos , Sirolimo/uso terapêutico , SolubilidadeRESUMO
Atrazine is an herbicide commonly used in several countries. Due to its long half-life, associated with its use in large scales, atrazine residues remain as environmental pollutants in water bodies. Phytoremediation is often pointed out as an interesting approach to remove atrazine from the aquatic environment, but its practical application is limited by the high toxicity of this herbicide. Here, we characterize the damages triggered by atrazine in Pistia stratiotes, evaluating the role of nitric oxide (NO), a cell-signaling molecule, in increasing the tolerance to the pollutant and the phytoremediation potential of this species. Pistia stratiotes plants were exposed to four treatments: Control; Sodium nitroprusside (SNP) (0.05 mg L-1); Atrazine (ATZ) (150 µg L-1) and ATZ + SNP. The plants remained under those conditions for 24 h for biochemical and physiological analysis and 3 days for the evaluation of relative growth rate. The presence of atrazine in plant cells triggered a series of biochemical and physiological damages, such as the increase in the generation of reactive oxygen species, damages to cell membranes, photosynthesis impairment, and negative carbon balance. Despite this, the plants maintained greater growth rates than other aquatic macrophytes exposed to atrazine and showed high bioconcentration and translocation factors. The addition of SNP, a NO donor, decreased the herbicide toxicity, with an increase of over 60% in the IC50 value (Inhibitor Concentration). Indeed, the NO signaling action was able to increase the tolerance of plants to atrazine, which resulted in increments in pollutant uptake and translocation, with the maintenance of overall cell (e.g. membranes) and organs (root system) structure, and the functioning of central physiological processes (e.g. photosynthesis). These factors allowed for more quickly and efficient removal of the pollutant from the environment, reducing costs, and increasing the viability of the phytoremediation process.
Assuntos
Araceae , Atrazina , Herbicidas , Poluentes Químicos da Água , Atrazina/toxicidade , Biodegradação Ambiental , Herbicidas/toxicidade , Óxido Nítrico , Água , Poluentes Químicos da Água/toxicidadeRESUMO
Rosmarinic acid, a plant-derived compound with antiangiogenic activity, can be applied for the treatment of ocular diseases related to neovascularization, such as diabetic retinopathy, macular edema, and age-related macular degeneration. These diseases represent the leading causes of blindness worldwide if they are not properly treated. Intravitreal devices allow for localized drug delivery to the posterior segment, increasing the drug bioavailability and promoting extended release, thus, reducing side effects and enhancing the patient's compliance to the treatment. In this work, rosmarinic acid-loaded poly lactic-co-glycolic acid intraocular implants were developed with a view for the treatment of ocular neovascularization. Physical-chemical, biocompatibility, and safety studies of the implants were carried out in vitro and in vivo as well as an evaluation of the antiangiogenic activity in a chorioallantoic membrane assay. Data obtained showed that rosmarinic acid released from the implants was quantified in the vitreous for 6 weeks, while when it was in the solution formulation, after 24 h, no drug was found in the vitreous. The delivery device did not show any sign of toxicity after clinical evaluation and in electroretinographic findings. Histological analysis showed normal eye tissue. Rosmarinic acid released from implants reduced 30% of new vessel's formation. The intravitreal implant successfully allowed for the prolonged release of rosmarinic acid, was safe to rabbits eyes, and demonstrated activity in vessel reduction, thus demonstrating potential in preventing neovascularization in ophthalmic diseases.
Assuntos
Depsídeos , Corpo Vítreo , Animais , Cinamatos , Depsídeos/farmacologia , Humanos , Injeções Intravítreas , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Coelhos , Ácido RosmarínicoRESUMO
During shoot development, leaves undergo various ontogenetic changes, including variation in size, shape, and geometry. Passiflora edulis (passionfruit) is a heteroblastic species, which means that it experiences conspicuous changes throughout development, enabling a morphological distinction between the juvenile and adult vegetative phases. Quantification of heteroblasty requires a practical, inexpensive, reliable, and non-destructive method, such as remote sensing. Moreover, relationships among ontogenetic changes and spectral signal at leaf level can be scaled up to support precision agriculture in passion fruit crops. In the present study, we used laboratory spectroscopic measurements (400-2500 nm) and narrowband vegetation indexes (or hyperspectral vegetation indexes - HVIs) to evaluate ontogenetic changes related to development and aging in P. edulis leaves. We also assessed leaf pigment concentration to further support the application of biochemical-related narrowband indexes. We report that 30-d-old leaves can be discriminated into developmental stages through their spectral signals. MSI (Moisture Stress Index) and NDVI750 (Normalized Difference Vegetation Index ρ750) contribute most to the variation of age (15 to 30-d-old leaves) and developmental stage (phytomer positions along the plant axis) in passionfruit leaves. PRI (Photochemical Reflectance Index) played an important role in detecting age and development alterations, including heteroblasty. A biochemical and spectral comparison of pigments revealed that spectroscopy offered potential for diagnosing phenology in P. edulis, as some narrowband indexes correlated strongly with chlorophylls and carotenoids content. Narrowband vegetation indexes are found to be a suitable tool for monitoring passionfruit crops.
Assuntos
Passiflora/crescimento & desenvolvimento , Folhas de Planta/crescimento & desenvolvimento , Análise Espectral/métodos , Carotenoides/análise , Clorofila/análiseRESUMO
Temporomandibular disorder is a clinical painful condition in the temporomandibular joint (TMJ) region. The purified sulfated polysaccharide from the green marine algae Caulerpa racemosa (Cr) has provided anti-inflammatory and antinociceptive activity. This study evaluated these effects on a TMJ hypernociception model. Wistar rats (180 - 250 g) were pre-treated (i.v.) with Cr at 0.01, 0.1, or 1 mg/kg or vehicle 30 min before formalin (1.5%/50 µL, i.art.), capsaicin (1.5%/20 µL, i.art.), or serotonin (225 µg/50 µL, i.art.) in the TMJ, and nociceptive behaviors were measured for 45 or 30 min upon inflammatory stimuli. Inflammatory parameters vascular permeability assay, TNF-α, and IL-1ß by ELISA, protein expression of adhesion molecules ICAM-1 and CD55 by Western blot were assessed. The involvement of heme oxygenase-1 (HO-1) and nitric oxide (NO) pathways were assessed by pharmacological inhibition. Cr (1 mg/kg) reduced nociceptive behavior, plasmatic extravasation, TNF-α, and IL-1ß levels, as well as ICAM-1 and CD55 expression in periarticular tissues. Cr antinociceptive effect was not prevented by aminoguanidine, but ZnPP-IX did reduce its antinociceptive effect. Therefore, Cr antinociceptive and anti-inflammatory effects in this experimental model of hypernociception depended on the HO-1 pathway integrity, as well as reducing peripheral inflammatory events, e.g., TNF-α and IL-1ß cytokines levels, ICAM-1 and CD55 expression.
Assuntos
Analgésicos/química , Analgésicos/farmacologia , Organismos Aquáticos/química , Clorófitas/química , Polissacarídeos/química , Polissacarídeos/farmacologia , Sulfatos/química , Animais , Artralgia/tratamento farmacológico , Artralgia/etiologia , Artralgia/metabolismo , Biomarcadores , Capsaicina/efeitos adversos , Citocinas/metabolismo , Heme Oxigenase-1/metabolismo , Mediadores da Inflamação/metabolismo , Masculino , Óxido Nítrico/metabolismo , Ratos , Articulação Temporomandibular/efeitos dos fármacos , Articulação Temporomandibular/fisiopatologiaRESUMO
Uveitis encompasses a heterogeneous and complex group of conditions characterized by intraocular inflammation, frequently affecting young individuals and representing an important cause of irreversible blindness worldwide. Animal models have been critical to understand etiology and pathogenesis of uveitis, being also employed to assess new therapeutic strategies, preceding human studies. However, there is still a need of developing and studying different models, due to the difficulties in recapitulating all forms of human uveitis effectively. Although corticosteroids are usually the first-line therapy for non-infectious uveitis, their long-term use is limited by potentially serious side effects in all possible delivery routes. Thus, thalidomide, a drug with anti-inflammatory and antiangiogenic properties, was investigated in a novel experimental model of uveitis, induced by Mycobacterium bovis Calmette-Guérin Bacillus (BCG), in rabbits. The experimental protocol consisted of two subcutaneous injections of BCG, followed by two intravitreal injections of the same antigen, inducing panuveitis. Animals were treated with a single intravitreal injection of thalidomide suspension or PBS. Clinical manifestations of uveitis improved after intravitreal thalidomide, involving both anterior and posterior segments. Protein content, N-acetyl-b-glucosaminidase (NAG) and myeloperoxidase (MPO) activities were elevated in ocular tissues after disease induction, further decreasing post-treatment with intravitreal thalidomide. This therapeutic response was also confirmed on ocular electrophysiology, as well as histopathology. This experimental model induced panuveitis in rabbits using a low-cost mycobacterial antigen, with intraocular inflammation subsequently improving after treatment. Intravitreal thalidomide may be a potential alternative to treat intraocular inflammation in corticosteroid-sparing therapies.
Assuntos
Anti-Inflamatórios/uso terapêutico , Doenças Autoimunes/tratamento farmacológico , Pan-Uveíte/tratamento farmacológico , Retina/metabolismo , Talidomida/uso terapêutico , Uveíte/tratamento farmacológico , Animais , Humanos , Injeções Intravítreas , Modelos Animais , Mycobacterium bovis/imunologia , Pan-Uveíte/imunologia , Peroxidase/metabolismo , Coelhos , Retina/efeitos dos fármacos , Retina/patologiaRESUMO
BACKGROUND AND AIMS: Juvenile-to-adult phase transition is marked by changes in leaf morphology, mostly due to the temporal development of the shoot apical meristem, a phenomenon known as heteroblasty. Sugars and microRNA-controlled modules are components of the heteroblastic process in Arabidopsis thaliana leaves. However, our understanding about their roles during phase-changing in other species, such as Passiflora edulis, remains limited. Unlike Arabidopsis, P. edulis (a semi-woody perennial climbing vine) undergoes remarkable changes in leaf morphology throughout juvenile-to-adult transition. Nonetheless, the underlying molecular mechanisms are unknown. METHODS: Here we evaluated the molecular mechanisms underlying the heteroblastic process by analysing the temporal expression of microRNAs and targets in leaves as well as the leaf metabolome during P. edulis development. KEY RESULTS: Metabolic profiling revealed a unique composition of metabolites associated with leaf heteroblasty. Increasing levels of glucose and α-trehalose were observed during juvenile-to-adult phase transition. Accumulation of microRNA156 (miR156) correlated with juvenile leaf traits, whilst miR172 transcript accumulation was associated with leaf adult traits. Importantly, glucose may mediate adult leaf characteristics during de novo shoot organogenesis by modulating miR156-targeted PeSPL9 expression levels at early stages of shoot development. CONCLUSIONS: Altogether, our results suggest that specific sugars may act as co-regulators, along with two microRNAs, leading to leaf morphological modifications throughout juvenile-to-adult phase transition in P. edulis.
Assuntos
Arabidopsis , MicroRNAs , Passiflora , Regulação da Expressão Gênica de Plantas , Folhas de PlantaRESUMO
A new species of Pyrrhulina is described based on 60 specimens from tributaries of the rio Amazonas: rio Anapu, rio Capim, rio Guamá and rio Xingu, and coastal drainages in the state of Pará, Brazil. The new species differs from all congeners by having the primary stripe (the horizontal stripe of dark pigmentation extending posteriorly from the snout) terminating at the distal edge of the opercle. In all other species of Pyrrhulina, the primary stripe is either restricted to the snout or continues beyond the head (i.e. at least the anteriormost four scales of the lateral line series).(AU)
Uma nova espécie de Pyrrhulina é descrita baseada em 60 espécimes dos afluentes do rio Amazonas: rio Anapu, rio Capim, rio Guamá e rio Xingu, e drenagens costeiras no estado do Pará, Brasil. A nova espécie distingue-se de todas as congêneres por ter a faixa primária (faixa horizontal de pigmentação escura que se estende posteriormente do focinho) terminando na borda distal do opérculo. Em todas as outras espécies de Pyrrhulina, a faixa primária é restrita ao focinho ou continua além da cabeça (para incluir pelo menos as quatro escamas anteriores da série da linha lateral).(AU)
Assuntos
Animais , Caraciformes/anatomia & histologia , Caraciformes/classificação , Caraciformes/crescimento & desenvolvimentoRESUMO
The bioconversion of protein from the fishing industry waste into bioplastics allows the valorization of biological material, reducing the production of waste and consequently of negative environmental impacts through the use of synthetic packaging. The objective of this work was to develop biodegradable films from the mixture of gelatin and myofibrillar fish proteins. Proteins (myofibrillar and gelatin) were extracted from filleting residues from king weakfish (Macrodon ancylodon) from industrial fishing. The properties of the blend films were compared to those of individual protein films. It was found by scanning electron microscopy that there was good compatibility between the two polymers. Mechanical analyzes showed that myofibrillar proteins films were the most resistant to traction, but less flexible; characteristics contrary to those of gelatin. The mixing films presented the lowest values of water vapor permeability and solubility were transparent, and mechanically strong and flexible, confirming the improvement of the properties by mixing the polymers. Fourier transform infrared spectroscopy revealed that there was interaction between the myofibrillar protein chains and gelatine driven by hydrogen bonds, thus forming cohesive and reinforced matrix, which resulted in good thermal resistance of the films. The mixture between polymers improved the technological properties of the elaborated biodegradable films, making its application feasible as food packaging.
A bioconversão de proteínas de subprodutos da indústria pesqueira em bioplástico possibilita a valorização do material biológico, reduzindo a produção de descarte e consequentemente, de impactos ambientais negativos do uso de embalagens sintéticas. O objetivo deste trabalho foi misturar gelatina e proteínas miofibrilares de pescada gó (Macrodon ancylodon) com intuito de desenvolver filmes biodegradáveis com propriedades tecnológicas aprimoradas. As propriedades dos filmes de mistura foram comparadas com as de filmes dos polímeros individuais. A espectroscopia de infravermelho com transformada de Fourier revelou boa compatilidade química entre gelatina e proteínas miofibrilares na formação do filme de mistura. A interação entre os polímeros foi conduzida por ligações de hidrogênio, formando película coesa e reforçada. Os filmes de mistura além de apresentarem os menores valores de permeabilidade ao vapor de água e solubilidade, também foram transparentes e, termicamente resistentes. Assim a mistura entre gelatina e proteínas miofibrilares possibilitou a formação de películas biodegradáveis com propriedades tecnológicas superiores às produzidas com os polímeros separados.
Assuntos
Animais , Gelatina , Miofibrilas , Peixes , Polímeros , Resíduos de AlimentosRESUMO
A new species of Pyrrhulina is described based on 60 specimens from tributaries of the rio Amazonas: rio Anapu, rio Capim, rio Guamá and rio Xingu, and coastal drainages in the state of Pará, Brazil. The new species differs from all congeners by having the primary stripe (the horizontal stripe of dark pigmentation extending posteriorly from the snout) terminating at the distal edge of the opercle. In all other species of Pyrrhulina, the primary stripe is either restricted to the snout or continues beyond the head (i.e. at least the anteriormost four scales of the lateral line series).(AU)
Uma nova espécie de Pyrrhulina é descrita baseada em 60 espécimes dos afluentes do rio Amazonas: rio Anapu, rio Capim, rio Guamá e rio Xingu, e drenagens costeiras no estado do Pará, Brasil. A nova espécie distingue-se de todas as congêneres por ter a faixa primária (faixa horizontal de pigmentação escura que se estende posteriormente do focinho) terminando na borda distal do opérculo. Em todas as outras espécies de Pyrrhulina, a faixa primária é restrita ao focinho ou continua além da cabeça (para incluir pelo menos as quatro escamas anteriores da série da linha lateral).(AU)
Assuntos
Animais , Caraciformes/anatomia & histologia , Caraciformes/classificação , Caraciformes/crescimento & desenvolvimentoRESUMO
The bioconversion of protein from the fishing industry waste into bioplastics allows the valorization of biological material, reducing the production of waste and consequently of negative environmental impacts through the use of synthetic packaging. The objective of this work was to develop biodegradable films from the mixture of gelatin and myofibrillar fish proteins. Proteins (myofibrillar and gelatin) were extracted from filleting residues from king weakfish (Macrodon ancylodon) from industrial fishing. The properties of the blend films were compared to those of individual protein films. It was found by scanning electron microscopy that there was good compatibility between the two polymers. Mechanical analyzes showed that myofibrillar proteins films were the most resistant to traction, but less flexible; characteristics contrary to those of gelatin. The mixing films presented the lowest values of water vapor permeability and solubility were transparent, and mechanically strong and flexible, confirming the improvement of the properties by mixing the polymers. Fourier transform infrared spectroscopy revealed that there was interaction between the myofibrillar protein chains and gelatine driven by hydrogen bonds, thus forming cohesive and reinforced matrix, which resulted in good thermal resistance of the films. The mixture between polymers improved the technological properties of the elaborated biodegradable films, making its application feasible as food packaging.(AU)
A bioconversão de proteínas de subprodutos da indústria pesqueira em bioplástico possibilita a valorização do material biológico, reduzindo a produção de descarte e consequentemente, de impactos ambientais negativos do uso de embalagens sintéticas. O objetivo deste trabalho foi misturar gelatina e proteínas miofibrilares de pescada gó (Macrodon ancylodon) com intuito de desenvolver filmes biodegradáveis com propriedades tecnológicas aprimoradas. As propriedades dos filmes de mistura foram comparadas com as de filmes dos polímeros individuais. A espectroscopia de infravermelho com transformada de Fourier revelou boa compatilidade química entre gelatina e proteínas miofibrilares na formação do filme de mistura. A interação entre os polímeros foi conduzida por ligações de hidrogênio, formando película coesa e reforçada. Os filmes de mistura além de apresentarem os menores valores de permeabilidade ao vapor de água e solubilidade, também foram transparentes e, termicamente resistentes. Assim a mistura entre gelatina e proteínas miofibrilares possibilitou a formação de películas biodegradáveis com propriedades tecnológicas superiores às produzidas com os polímeros separados.(AU)
Assuntos
Animais , Polímeros , Peixes , Miofibrilas , Gelatina , Resíduos de AlimentosRESUMO
The [18F]Fluorocholine ([18F]FCH) tracer for PET imaging has been proven to be effective for several malignances. However, there are only a few studies related to its breast tumor applicability and they are still limited. The aim of this study was investigate the efficacy of [18F]FCH/PET compared to [18F]FDG/PET in a murine 4T1 mammary carcinoma model treated and nontreated. [18F]FCH/PET showed its applicability for primary tumor and lung metastasis detection and their use for response monitoring of breast cancer therapeutics at earlier stages.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Neoplasias Mamárias Experimentais/diagnóstico por imagem , Animais , Antineoplásicos/uso terapêutico , Neoplasias da Mama/patologia , Neoplasias da Mama/terapia , Linhagem Celular Tumoral , Colina/análogos & derivados , Cinamatos/uso terapêutico , Depsídeos/uso terapêutico , Progressão da Doença , Feminino , Radioisótopos de Flúor , Fluordesoxiglucose F18 , Humanos , Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/secundário , Neoplasias Mamárias Experimentais/patologia , Neoplasias Mamárias Experimentais/terapia , Camundongos , Camundongos Endogâmicos BALB C , Tomografia por Emissão de Pósitrons/métodos , Compostos Radiofarmacêuticos , Ácido RosmarínicoRESUMO
In vitro plant regeneration systems have turned into invaluable tools to plant biotechnology. Despite being poorly understood, the molecular mechanisms underlying the control of both morphogenetic pathways, de novo organogenesis and somatic embryogenesis, have been supported by recent findings involving proteome-, metabolome-, and transcriptome-based profiles. Notwithstanding, the integration of molecular data with structural aspects has been an important strategy of study attempting to elucidate the basis of the cell competence acquisition to further follow commitment and determination to specific a particular in vitro regeneration pathway. In that sense, morpho-histological tools have allowed to recognize cellular markers and patterns of gene expression at cellular level and this way have collaborated in the identification of the cell types with high regenerative capacity. This chapter ties together up those fundamental and important microscopy techniques that help to elucidate that regeneration occurs, most of the time, from epidermis or subepidermal cells and from the procambial cells (pericycle and vascular parenchyma). Important findings are discussed toward ultrastructural differences observed in the nuclear organization among pluripotent and totipotent cells, implying that regeneration occurs from two cellular mechanisms based on cellular reprogramming or reactivation.
Assuntos
Células Vegetais/metabolismo , Plantas/anatomia & histologia , Regeneração , Hibridização In Situ , Células Vegetais/ultraestrutura , Plantas/ultraestruturaRESUMO
Temporomandibular disorders are the second most common cause of orofacial pain mediated by inflammatory compounds, which in many cases leads to chronic orofacial pain. This study assessed the antinociceptive and anti-inflammatory effects of a lectin from the green seaweed Caulerpa cupressoides (CcL) on hypernociception inflammatory in TMJ of rats and investigated the involvement of different mechanisms. Rats received i.v. CcL 30â¯min prior to injection of flogistic agentes or 0.9% saline into the left TMJ. Pretreatment with CcL (0. 1; 1 or 10â¯mg/kg) promoted a reduction (pâ¯<â¯0.05) of inflammatory hypernociception induced by 1.5% Formalin along with inhibition of inflammatory plasma extravasation, cytokines levels, ciclooxigenase-2, and intercellular adhesion molecule (ICAM-1). CcL was able to inhibit the nociceptive response induced by 1.5% Capsaicin, suggesting that CcL has an antinociceptive effect, acting directly on the primary nociceptive neurons. CcL also inhibited the nociceptive response induced by Carrageenan (100⯵g/TMJ) or Serotonin (5-HT) (225⯵g/TMJ). In conclusion, the results demonstrate that administration of CcL has a potential antinociceptive and anti-inflammatory effect, with a mechanism that is partially dependent on TNF-α, IL-1ß, COX-2 and ICAM-1 inhibition and independently from the cannabinoide and opioid system and NO/cGMP/PKG/K+ATP channel pathway.
Assuntos
Analgésicos/farmacologia , Caulerpa/química , Lectinas de Plantas/farmacologia , Articulação Temporomandibular/efeitos dos fármacos , Animais , Moléculas de Adesão Celular/metabolismo , Ciclo-Oxigenase 2/metabolismo , Inflamação/fisiopatologia , Interleucina-1beta/biossíntese , Masculino , Atividade Motora/efeitos dos fármacos , Nociceptividade/efeitos dos fármacos , Ratos , Ratos Wistar , Articulação Temporomandibular/metabolismo , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Abelmoschus esculentus is largely cultivated in Northeastern Brazil for medicinal purposes, e.g. inflammatory conditions. This study aimed to evaluate the efficacy of Abelmoschus esculentus lectin (AEL) in reducing formalin-induced temporomandibular joint inflammatory hypernociception in rats. The behavioral experiments were performed in male Wistar rats (180-240â¯g). Rats were pre-treated (i.v.) with AEL (0.001, 0.01 or 0.1â¯mg/kg) 30â¯min before formalin injection (i.art.). To analyze the possible effect of opioid pathways on AEL efficacy, animals were pre-treated with naloxone or CTOP (µ opioid receptor antagonist), naltrindole (δ opioid receptor antagonist) or nor-binaltorphimine (κ opioid receptor antagonist) (i.t.) 15â¯min before AEL administration followed by intra-TMJ injection of 1.5% formalin. Animals were monitored for a 45-min observation period. TMJ tissue, trigeminal ganglion, and subnucleus caudalis were collected for TNF-α dosage (ELISA). In addition, the vascular permeability was evaluated by Evans Blue extravasation. AEL significantly reduced formalin-induced TMJ inflammatory hypernociception and decreased Evans blue extravasation. It decreased TNF-α levels in the TMJ tissue, trigeminal ganglion, and subnucleus caudalis. AEL antinociceptive effects were not observed in the presence of naltrindole or nor-binaltorphimine, suggesting that AEL efficacy depends on TNF-α inhibition and the activation of δ and κ opioid receptors. AEL has provided prominent analgesic and anti-inflammatory effects in this pre-clinical model of TMJ, supporting its possible use as a pharmacological tool for the management of painful conditions.
Assuntos
Abelmoschus/química , Analgésicos/farmacologia , Lectinas/farmacologia , Dor/tratamento farmacológico , Receptores Opioides/metabolismo , Articulação Temporomandibular/efeitos dos fármacos , Analgésicos Opioides/farmacologia , Animais , Anti-Inflamatórios/farmacologia , Formaldeído/farmacologia , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Masculino , Hipernutrição/tratamento farmacológico , Hipernutrição/metabolismo , Dor/induzido quimicamente , Dor/metabolismo , Ratos , Ratos Wistar , Articulação Temporomandibular/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
This study aimed to develop and characterize biodegradable films from myofibrillar proteins from whitemouth croaker (Micropogonias furnieri) using stearic acid (SA) to improve its technological properties. The proteins extracted were lyophilized, characterized, and used to prepare the films. The filmogenic solutions were dried on a silicone stand at 35 °C for 14 h in an incubator oven according to the casting method. The region with the best mechanical, physical, and barrier properties in the films was defined by a full factorial design. The lyophilized myofibrillar proteins (LMP) had protein content of 96.03%. The analysis of the experimental design results indicated the best conditions to prepare the optimized film were 2.84% LMP, 3.18% SA, and 78.41% SDS (sodium dodecyl sulfate). The control films were prepared with 2.84% LMP and 30% plasticizer. The optimized film had significantly lower water vapor permeability (5.87E-11 g m m-2 s-1 Pa-1), higher tensile strength (6.35 MPa), and lower elongation (235.60%) compared with control (p≤0.05). It also had lower transparency values and excellent UV barrier property, which indicates a tendency to opaque. Thermal stability was good and the microstructure revealed a structural change in the filmogenic matrix, confirmed by x-ray diffraction, which indicates influence of SDS and SA on film crystallinity. Solubility increased by 22% and swelling decreased slightly in the optimized film compared to the control. The results obtained represent a positive contribution with the use of fish byproducts by applying alternative, sustainable technologies.(AU)
O objetivo deste trabalho foi desenvolver e caracterizar filmes biodegradáveis de proteínas miofibrilares de aparas da filetagem da corvina (Micropogonias furnieri) utilizando ácido esteárico (AE) para melhorar suas propriedades tecnológicas. As proteínas extraídas foram liofilizadas, caracterizadas e utilizadas na elaboração dos filmes. As soluções filmogênicas foram secas em suporte de silicone a 35°C por 14 horas em estufa incubadora, de acordo com o método casting. A região com as melhores propriedades mecânicas, físicas e de barreira dos filmes foi definida por planejamento fatorial completo. As proteínas miofibrilares liofilizadas (PML) apresentaram teor proteico de 96,03%. A análise dos resultados do planejamento experimental indicou que as melhores condições para elaborar o filme otimizado foram: 2,84% PML, 3,18% AE e 78,41% SDS (dodecil sulfato de sódio). Os filmes controles foram elaborados com 2,84% de PML e 30% de plastificante. O filme otimizado apresentou significativa diminuição da permeabilidade ao vapor de água (5,87E-11 g m m-2 s-1 Pa-1), com maior resistência a tração (6,35 MPa) e menor elongação (235,60%) quando comparado ao controle (p ≤ 0,05. Apresentando também menores valores de transparência e excelente propriedade de barreira UV, indicando tendência ao opaco. Apresentou boa estabilidade térmica e a microestrutura revelou mudança estrutural na matriz filmogênica, confirmado pela difração de raio-x, indicando a influência do SDS e AE na cristalinidade do filme. Houve aumento de 22% na solubilidade e ligeira diminuição do intumescimento do filme otimizado em relação ao controle. Os resultados obtidos representam contribuição positiva com o aproveitamento de subprodutos de peixe aplicando tecnologias alternativas e sustentáveis.(AU)
Assuntos
Animais , Técnica de Nutrientes em Filmes/métodos , Miofibrilas , Ácidos Esteáricos , Dodecilsulfato de Sódio , Plásticos Biodegradáveis/análise , Proteínas de Peixes , PerciformesRESUMO
This study aimed to develop and characterize biodegradable films from myofibrillar proteins from whitemouth croaker (Micropogonias furnieri) using stearic acid (SA) to improve its technological properties. The proteins extracted were lyophilized, characterized, and used to prepare the films. The filmogenic solutions were dried on a silicone stand at 35 °C for 14 h in an incubator oven according to the casting method. The region with the best mechanical, physical, and barrier properties in the films was defined by a full factorial design. The lyophilized myofibrillar proteins (LMP) had protein content of 96.03%. The analysis of the experimental design results indicated the best conditions to prepare the optimized film were 2.84% LMP, 3.18% SA, and 78.41% SDS (sodium dodecyl sulfate). The control films were prepared with 2.84% LMP and 30% plasticizer. The optimized film had significantly lower water vapor permeability (5.87E-11 g m m-2 s-1 Pa-1), higher tensile strength (6.35 MPa), and lower elongation (235.60%) compared with control (p≤0.05). It also had lower transparency values and excellent UV barrier property, which indicates a tendency to opaque. Thermal stability was good and the microstructure revealed a structural change in the filmogenic matrix, confirmed by x-ray diffraction, which indicates influence of SDS and SA on film crystallinity. Solubility increased by 22% and swelling decreased slightly in the optimized film compared to the control. The results obtained represent a positive contribution with the use of fish byproducts by applying alternative, sustainable technologies.
O objetivo deste trabalho foi desenvolver e caracterizar filmes biodegradáveis de proteínas miofibrilares de aparas da filetagem da corvina (Micropogonias furnieri) utilizando ácido esteárico (AE) para melhorar suas propriedades tecnológicas. As proteínas extraídas foram liofilizadas, caracterizadas e utilizadas na elaboração dos filmes. As soluções filmogênicas foram secas em suporte de silicone a 35°C por 14 horas em estufa incubadora, de acordo com o método casting. A região com as melhores propriedades mecânicas, físicas e de barreira dos filmes foi definida por planejamento fatorial completo. As proteínas miofibrilares liofilizadas (PML) apresentaram teor proteico de 96,03%. A análise dos resultados do planejamento experimental indicou que as melhores condições para elaborar o filme otimizado foram: 2,84% PML, 3,18% AE e 78,41% SDS (dodecil sulfato de sódio). Os filmes controles foram elaborados com 2,84% de PML e 30% de plastificante. O filme otimizado apresentou significativa diminuição da permeabilidade ao vapor de água (5,87E-11 g m m-2 s-1 Pa-1), com maior resistência a tração (6,35 MPa) e menor elongação (235,60%) quando comparado ao controle (p ≤ 0,05. Apresentando também menores valores de transparência e excelente propriedade de barreira UV, indicando tendência ao opaco. Apresentou boa estabilidade térmica e a microestrutura revelou mudança estrutural na matriz filmogênica, confirmado pela difração de raio-x, indicando a influência do SDS e AE na cristalinidade do filme. Houve aumento de 22% na solubilidade e ligeira diminuição do intumescimento do filme otimizado em relação ao controle. Os resultados obtidos representam contribuição positiva com o aproveitamento de subprodutos de peixe aplicando tecnologias alternativas e sustentáveis.
Assuntos
Animais , Dodecilsulfato de Sódio , Miofibrilas , Plásticos Biodegradáveis/análise , Técnica de Nutrientes em Filmes/métodos , Ácidos Esteáricos , Perciformes , Proteínas de PeixesRESUMO
The wild grass species Brachypodium distachyon (L.) has been proposed as a new model for temperate grasses. Among the biotechnological tools already developed for the species, an efficient induction protocol of somatic embryogenesis (SE) using immature zygotic embryos has provided the basis for genetic transformation studies. However, a systematic work to better understanding the basic cellular and molecular mechanisms that underlie the SE process of this grass species is still missing. Here, we present new insights at the morpho-histological, histochemical, and molecular aspects of B. distachyon SE pathway. Somatic embryos arose from embryogenic callus formed by cells derived from the protodermal-dividing cells of the scutellum. These protodermal cells showed typical meristematic features and high protein accumulation which were interpreted as the first observable steps towards the acquisition of a competent state. Starch content decreased along embryogenic callus differentiation supporting the idea that carbohydrate reserves are essential to morphogenetic processes. Interestingly, starch accumulation was also observed at late stages of SE process. Searches in databanks revealed three sequences available annotated as BdSERK, being two copies corresponding to SERK1 and one showing greater identity to SERK2. In silico analysis confirmed the presence of characteristic domains in a B. distachyon Somatic Embryogenesis Receptor Kinase genes candidates (BdSERKs), which suggests SERK functions are conserved in B. distachyon. In situ hybridization demonstrated the presence of transcripts of BdSERK1 in all development since globular until scutellar stages. The results reported in this study convey important information about the morphogenetic events in the embryogenic pathway which has been lacking in B. distachyon. This study also demonstrates that B. distachyon provides a useful model system for investigating the genetic regulation of SE in grass species.