RESUMO
The peptide derived from E. contortisiliquum trypsin inhibitor (Pep-3-EcTI), peptide derived from kallikrein inhibitor isolated from B. bauhinioides (Pep-BbKI), and B. rufa peptide modified from B. bauhinioides (Pep-BrTI) peptides exhibit anti-inflammatory and antioxidant activities, suggesting their potential for treating asthma-chronic obstructive pulmonary disease (COPD) overlap (ACO). We compared the effects of these peptides with dexamethasone (DX) treatment in an ACO model. In this study, 11 groups of male BALB/c mice were pre-treated under different conditions, including sensitization with intraperitoneal injection and inhalation of ovalbumin (OVA), intratracheal instillation of porcine pancreatic elastase (ELA), sensitization with intraperitoneal injection, and various combinations of peptide treatments with Pep-3-EcTI, Pep-BbKI, Pep-BrTI, dexamethasone, and non-treated controls (SAL-saline). Respiratory system resistance, airway resistance, lung tissue resistance, exhaled nitric oxide, linear mean intercept, immune cell counts in the bronchoalveolar lavage fluid, cytokine expression, extracellular matrix remodeling, and oxidative stress in the airways and alveolar septa were evaluated on day 28. Results showed increased respiratory parameters, inflammatory markers, and tissue remodeling in the ACO group compared to controls. Treatment with the peptides or DX attenuated or reversed these responses, with the peptides showing effectiveness in controlling hyperresponsiveness, inflammation, remodeling, and oxidative stress markers. These peptides demonstrated an efficacy comparable to that of corticosteroids in the ACO model. However, this study highlights the need for further research to assess their safety, mechanisms of action, and potential translation to clinical studies before considering these peptides for human use.
RESUMO
(1) There are several patients with asthma-COPD overlap (ACO). A peptide derived from the primary sequence of a kallikrein inhibitor isolated from Bauhinia bauhinioides (pep-BbKI) has potent anti-inflammatory and antioxidant effects. Purpose: To investigate the effects of pep-BbKI treatment in an ACO model and compare them with those of corticosteroids. (2) BALB/c mice were divided into groups: SAL (saline), OVA (ovalbumin), ELA (elastase), ACO (ovalbumin + elastase), ACO-pep-BbKI (treated with inhibitor), ACO-DX (dexamethasone treatment), ACO-DX-pep-BbKI (both treatments), and SAL-pep-BbKI (saline group treated with inhibitor). We evaluated: hyperresponsiveness to methacholine, bronchoalveolar lavage fluid (BALF), exhaled nitric oxide (eNO), IL-1ß, IL-4, IL-5, IL-6, IL-10, IL-13, IL-17, IFN-γ, TNF-α, MMP-9, MMP-12, TGF-ß, collagen fibers, iNOS, eNO, linear mean intercept (Lm), and NF-κB in airways (AW) and alveolar septa (AS). (3) ACO-pep-BbKI reversed ACO alterations and was similar to SAL in all mechanical parameters, Lm, neutrophils, IL-5, IL-10, IL-17, IFN-γ, TNF-α, MMP-12 (AW), collagen fibers, iNOS (AW), and eNO (p > 0.05). ACO-DX reversed ACO alterations and was similar to SAL in all mechanical parameters, Lm, total cells and differentials, IL-1ß(AS), IL-5 (AS), IL-6 (AS), IL-10 (AS), IL-13 (AS), IFN-γ, MMP-12 (AS), TGF-ß (AS), collagen fibers (AW), iNOS, and eNO (p > 0.05). SAL was similar to SAL-pep-BbKI for all comparisons (p > 0.05). (4) Pep-BbKI was similar to dexamethasone in reducing the majority of alterations of this ACO model.
Assuntos
Asma , Bauhinia , Doença Pulmonar Obstrutiva Crônica , Animais , Camundongos , Interleucina-10 , Interleucina-17 , Ovalbumina , Interleucina-13 , Interleucina-5 , Interleucina-6 , Metaloproteinase 12 da Matriz , Fator de Necrose Tumoral alfa , Proteínas de Plantas/farmacologia , Peptídeos/farmacologia , Líquido da Lavagem Broncoalveolar , Asma/tratamento farmacológico , Calicreínas , Elastase Pancreática , Dexametasona , Colágeno , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Modelos Animais de Doenças , Camundongos Endogâmicos BALB CRESUMO
Introduction: Small airways are not evaluated with traditional pulmonary function tests. The aim of this study was to evaluate the small airways in patients with chronic obstructive pulmonary disease (COPD) with a nitrogen washout test and to verify whether there is a difference between patients with COPD due to smoking and those with COPD due to alpha-1 antitrypsin mutation. Methods: Sixteen patients with mutation in the SERPINA1 gene and 45 patients with no mutation were included in this cross-sectional study. All pulmonary function tests, including the single breath nitrogen washout test, were performed for all patients and alpha-1 antitrypsin dosage was assessed with immunonephelometry. Results: A comparison of patients with COPD due to smoking and those with COPD due to smoking and mutation revealed a significant difference in closure volume (%), which was the poorest in the mutation group. In the group with COPD and mutation, there was an inverse correlation between smoking and closure volume (%). We also verified that similar to forced expiratory volume in the first second (FEV1), the phase III slope (%) and ΔN2 750-1250 mL (%) could be used to differentiate the severity of airflow limitation. Conclusion: Our results suggest that both variables, phase III slope and the ΔN2 750-1250 mL (%), could be related to COPD severity. Therefore, alterations at the distribution of the location of the emphysema could alter the results of closer volume and that the nitrogen washout test is more sensitive when compared to traditional pulmonary function test in evaluating COPD patients.
Assuntos
Doença Pulmonar Obstrutiva Crônica , Deficiência de alfa 1-Antitripsina , Idoso , Estudos Transversais , Volume Expiratório Forçado , Humanos , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Doença Pulmonar Obstrutiva Crônica/genética , Testes de Função Respiratória , alfa 1-Antitripsina , Deficiência de alfa 1-Antitripsina/complicações , Deficiência de alfa 1-Antitripsina/diagnóstico , Deficiência de alfa 1-Antitripsina/genéticaRESUMO
The recombinant TvCP4 prepro region (ppTvCP4r) acts as an exogenous inhibitor of cathepsin L-like CPs from Trichomonas vaginalis (Cárdenas-Guerra et al., 2015 [1]). Here, we present the dataset of the trichomonad ppTvCP4r inhibitory effect against the CP proteolytic activities from other microorganisms, such as Naegleria fowleri and Acanthamoeba castellanii free-living amoeba. The proteolytic activity inhibition of total crude extracts (TCEs) of N. fowleri and A. castellanii was determined and recorded using a fluorogenic substrate specific for cathepsin L CPs without or with a ppTvCP4r treatment at different concentrations and pH.
RESUMO
Plants respond to attacks by herbivores with various defences, which are mounted through the activation of different biochemical pathways that are known to interact. Thus, the attack of a plant by one herbivore species may result in changes in the performances of other species on the same plant. It has been suggested that species with comparable feeding modes induce similar plant defences and such herbivores are therefore expected to have a negative effect on each other's performance. We studied two closely related phytophagous mite species with identical feeding modes. Yet, one of the species (Tetranychus urticae) induces tomato plant defences, whereas the other (T. evansi) reduces them. We found that the "inducing" species benefits from the downregulation of defences by the "reducing" species, which, in turn, suffers from the induction of defences by the inducing species. Moreover, the performances of the two mite species on leaves that were previously attacked by both species simultaneously were intermediate between that on leaves previously attacked by each of the mites separately. The activity of proteinase inhibitor, a defensive compound, was not found to be intermediate in leaves attacked by both species simultaneously--it was almost as high as the activity seen in leaves with defences induced by T. urticae. Oviposition rates of T. urticae showed a nonlinear correlation with inhibitor activity, suggesting that it is potentially problematic to use this activity as an indicator of the level of plant defence. Our results show that herbivores with similar feeding modes have opposite effects on plant defence and differentially affect each other's performance on co-infested plants.
Assuntos
Comportamento Animal , Resistência à Doença , Herbivoria , Ácaros , Oviposição , Folhas de Planta/metabolismo , Solanum lycopersicum/fisiologia , Animais , Ecossistema , Feminino , Solanum lycopersicum/metabolismo , Doenças das Plantas , Inibidores de Proteases/metabolismo , Reprodução , TetranychidaeRESUMO
A prevalência de asma tem crescido e a maioria dos pacientes com asma grave não obtém o controle total dos sintomas com as terapias disponíveis, fazendo-se necessária a busca por novas alternativas terapêuticas. Inibidores de proteinases têm sido estudados como tratamento de processos inflamatórios, dentre eles o Enterolobium contortisiliquum Tripsin Inhibitor (EcTI) OBJETIVO: Avaliar se o inibidor de proteinase EcTI modula a hiperresponsividade brônquica à metacolina, inflamação, remodelamento e estresse oxidativo nas vias aéreas e septos alveolares em um modelo experimental de inflamação pulmonar alérgica crônica. MÉTODOS: Vinte e quatro camundongos Balb/c machos, entre seis e sete semanas de vida, pesando em media 25 g foram divididos em quatro grupos: C (controle), OVA (sensibilizados com ovalbumina, 50 ug intraperironeal (i.p) nos dias 0 e 14 e desafiados nos dias 22, 24, 26, 28); C+EC (controle tratados com EcTI (2 mg/kg/i.p) nos dias 22 a 28); OVA+EC (sensibilizados e desafiados com ovalbumina e também tratados com EcTI (2 mg/kg -i.p) nos dias 22 a 28). No dia 29, foram realizadas realizadas: (i) hiperresponsividade à metacolina e obtidas as respostas máximas de resistência e elastância do sistema respiratório; (ii) análise histopatológica do pulmão para quantificação de eosinófilos, fibras colágenas e elásticas nas vias aéreas (VA) e nos septos alveolares (SA); e (iii) imunohistoquímica para quantificação de células positivas para IFN-y, IL-4, IL-5, IL-13, MMP-9, TIMP-1, TGF-beta, iNOS, NF-kB e fração de volume de isoprostano nas VA e nos SA. Uma semana após o dia 29 foi realizada a técnica de anafilaxia cutanea passiva(PCA) para quantificar IgE e IgG1. A significância foi considerada quando p < 0,05. RESULTADOS: Houve aumento de todos os parâmetros avaliados no grupo OVA em relação ao grupo controle (p < 0,05). Houve atenuação da resposta máxima de Rrs e Ers no grupo OVA+EC comparado as grupo OVA (p < 0,05). O tratamento...
The number of cases of asthma has grown in recent decades. People who have severe asthma are likely to have more attacks and are at greater risk of a fatal attack, which propose to keep up global attention and keep approaching for advances in asthma care. Proteinase inhibitors of vegetable origin have been studied as a modulator of inflammatory responses and diseases. Among these inhibitors is Enterolobium contortisiliquum Trypsin Inhibitor (EcTI). AIMS: To evaluate the effects of EcTI in pulmonary mechanical, eosinophilic recruitment, inflammatory cytokines, remodeling of extracellular matrix and oxidative stressin an experimental model of chronic allergic pulmonary inflammation. METHODS: Twenty-four young adult male pathogen-free mice BALB/c (6-7 weeks old, 25-30g) were divided into 4 groups: C (control), OVA (sensitized with ovalbumin, 50 ug intraperitoneal (i.p), on days 0 and 14 and challenged with ova 1%, on days 22, 24, 26, 28); C+EC (control treated with EcTI- 2 mg/kg/i.p. from days 22 to 28); OVA+EC (sensitized and challenged with ovalbumin and treated with EcTI (2 mg/kg/i.p) from days 22 to 28). At day 29, we performed: (i) Bronchial hyperresponsiveness to methacholine and obtained the maximum response of resistance (Rrs) and elastance (Ers) of the respiratory system; (ii) lung histopathological analysis by morphometry to quantify eosinophils, collagen and elastic fibers volume fraction in airways; and (iii) immunohistochemistry to quantify IFN-y, IL-4, IL-5, IL-13, MMP-9, TIMP-1, TGF-, iNOS, NF-kB positive cells and isoprostane volume fraction in airways. One week after the day 29 we performed PCA technique to quantify IgE and IgG1 antibodies. Significance was considered at p < 0.05. RESULTS: The EcTI treatment in the ovalbumin-sensitized animals attenuated the maximal response of resistance and elastance of respiratory system after methacholine, the number of eosinophils, IL-4, IL-5, IL-13, IFN-y, NF-kB and iNOS-positive cells,...
Assuntos
Animais , Masculino , Camundongos , Remodelação das Vias Aéreas , Asma , Inflamação , Cloreto de Metacolina , Camundongos Endogâmicos BALB C , Estresse Oxidativo , Inibidores de Proteases , Inibidores da TripsinaRESUMO
Plants have a wide range of chemical defenses against predation, including substances that target digestive serine proteinases of herbivorous. Previous works demonstrated that lepidopteran insects have digestive serine proteinases resistant to plant proteinase inhibitors (PPIs) and ketone modifications, while coleopteran ones are sensitive to those plant defenses. This paper focuses on molecular aspects that lead lepidopteran serine proteinases to PPI and ketone modification resistance. Using biochemical experiments and computer 3D modeling we demonstrated that lepidopteran trypsins are more hydrophobic than coleopteran ones, a feature associated to trypsin oligomerization and decreased inhibition by PPI. Moreover, the determination of pKa values of chymotrypsin catalytic residues obtained by TPCK modification indicates that the environment around the active site of ketone-resistant and -sensitive chymotrypsins are different. Structural analysis using resistant and sensitive chymotrypsins data allowed us to point 2 hotspot regions around the active site that could explain the observed differences. Our set of results highlights features of serine proteinases important for understanding the resistance of insects to plant chemical defenses.
Assuntos
Lepidópteros/enzimologia , Fenômenos Fisiológicos Vegetais , Serina Proteases/metabolismo , Sequência de Aminoácidos , Animais , Modelos Moleculares , Dados de Sequência Molecular , Homologia de Sequência de Aminoácidos , Serina Proteases/química , Inibidores de Serina Proteinase/metabolismoRESUMO
Indolent ulcers are superficial corneal ulcers secondary to several changes on the corneal surface. They are frequently observed in middle-aged Boxer dogs, cause pain of acute onset and requires appropriate treatment. Aiming to evaluate the efficacy of clinical managements on the rate of healing of indolent ulcers, a retrospective study was conducted (1997-2008). Results demonstrated that proteinase inhibitors were the most often prescribed medication, and its administration did not interfere on the healing rate, as well as observed in dogs that received 1 percent atropine, antibiotics and anti-inflammatory drugs. Healing was delayed in dogs administered orally with vitamin C, but the healing process was faster on those dogs that went through corneal debridement/cauterization. In conclusion, to know the various types of treatments seems to be fundamental for the rapid resolution of the disease. It is suggested that debridement/cauterization, administration of proteinase inhibitor eye drops, prophylactic topical antibiotics and oral vitamin C, should be considered as an effective clinical management for indolent ulcers in Boxer dogs.
Úlceras indolentes são úlceras corneais superficiais e espontâneas, que apresentam curso prolongado e que tendem a recidivar. Comumente observadas em cães de meia idade da raça Boxer, provoca dor de início agudo e necessita de tratamento específico, já que este, quando não realizado corretamente, pode prolongar o curso da lesão por semanas a meses. Com o objetivo de avaliar a eficácia dos tratamentos clínicos quanto à rapidez na resolução do quadro, realizou-se estudo retrospectivo (1997 a 2008). Observou-se que os inibidores das proteinases foram as medicações mais frequentemente prescritas e que sua administração não interferiu no tempo de cicatrização corneal, o que também foi observado nos casos em que se administrou antibióticos e antinflamatórios tópicos e sistêmicos e/ou atropina 1 por cento. A administração de vitamina C retardou, de maneira estatisticamente significante, o tempo de cicatrização. Por outro lado, a realização do debridamento/cauterização corneal acelerou, significativamente, o processo. Conhecer os diversos tipos de tratamentos parece ser fundamental no sucesso e rapidez da resolução da doença. Os autores sugerem que a realização do debridamento/cauterização corneal, administração de inibidores das proteinases e antibióticos tópicos, associados à vitamina C por via oral, seja considerado um tratamento clínico efetivo na rápida resolução da doença.
Assuntos
Animais , Cães , Cauterização/veterinária , Diagnóstico Clínico/veterinária , Peptídeo Hidrolases , Úlcera da Córnea/veterinária , Estudos Retrospectivos , CicatrizaçãoRESUMO
Indolent ulcers are superficial corneal ulcers secondary to several changes on the corneal surface. They are frequently observed in middle-aged Boxer dogs, cause pain of acute onset and requires appropriate treatment. Aiming to evaluate the efficacy of clinical managements on the rate of healing of indolent ulcers, a retrospective study was conducted (1997-2008). Results demonstrated that proteinase inhibitors were the most often prescribed medication, and its administration did not interfere on the healing rate, as well as observed in dogs that received 1 percent atropine, antibiotics and anti-inflammatory drugs. Healing was delayed in dogs administered orally with vitamin C, but the healing process was faster on those dogs that went through corneal debridement/cauterization. In conclusion, to know the various types of treatments seems to be fundamental for the rapid resolution of the disease. It is suggested that debridement/cauterization, administration of proteinase inhibitor eye drops, prophylactic topical antibiotics and oral vitamin C, should be considered as an effective clinical management for indolent ulcers in Boxer dogs.(AU)
Úlceras indolentes são úlceras corneais superficiais e espontâneas, que apresentam curso prolongado e que tendem a recidivar. Comumente observadas em cães de meia idade da raça Boxer, provoca dor de início agudo e necessita de tratamento específico, já que este, quando não realizado corretamente, pode prolongar o curso da lesão por semanas a meses. Com o objetivo de avaliar a eficácia dos tratamentos clínicos quanto à rapidez na resolução do quadro, realizou-se estudo retrospectivo (1997 a 2008). Observou-se que os inibidores das proteinases foram as medicações mais frequentemente prescritas e que sua administração não interferiu no tempo de cicatrização corneal, o que também foi observado nos casos em que se administrou antibióticos e antinflamatórios tópicos e sistêmicos e/ou atropina 1 por cento. A administração de vitamina C retardou, de maneira estatisticamente significante, o tempo de cicatrização. Por outro lado, a realização do debridamento/cauterização corneal acelerou, significativamente, o processo. Conhecer os diversos tipos de tratamentos parece ser fundamental no sucesso e rapidez da resolução da doença. Os autores sugerem que a realização do debridamento/cauterização corneal, administração de inibidores das proteinases e antibióticos tópicos, associados à vitamina C por via oral, seja considerado um tratamento clínico efetivo na rápida resolução da doença.(AU)
Assuntos
Animais , Cães , Úlcera da Córnea/veterinária , Diagnóstico Clínico/veterinária , Cauterização/veterinária , Peptídeo Hidrolases , Estudos Retrospectivos , CicatrizaçãoRESUMO
The alpha2-macroglobulin (alpha2M) protease inhibitor was purified from bovine plasma. The alpha2M preparations at various purification steps were identified by immunodiffusion and crossed immunoelectrophoresis with anti-human alpha2 serum. Anti-bovine alpha2M serum was prepared for the quantitative determinations. The purest alpha2M preparation was obtained by affinity chromatography and used as primary standard in radial immunodiffusion. Alpha2M preparations were submitted to binding tests with p" - NPGB (p - nitrophenyl-p-guanidinebenzoate HCL) titrated trypsin and plasmin. Alpha2M protected 35% of the esterolytic activity of trypsin and 50% of the amidolytic activity of plasmin.
Alpha2 Macroglobulina, uma proteína inibidora de proteases, foi isolada do plasma bovino. O processo de purificação foi monitorado por imunodifusão e imunoeletroforese cruzada com soro anti alpha2M-humana. Para as determinações quantitativas foi preparado um soro anti alpha2M bovino. A preparação mais pura de alpha2M foi obtida por cromatografia de afinidade e usada como padrão primário na imunodifusão radial de Mancini. Preparações de alpha2M foram usadas em testes de ligação com tripsina e plasmina (tituladas com NPGB). Nos testes de ligação 50% de plasmina e 35% de tripsina foram "protegidas pela alpha2M. Não foi possível determinar se houve ineficiência na ligação ou se a perda de atividade ocorreu por alterações na afinidade do complexo alpha2M-protease, em relação aos substratos usados.