Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 3.153
Filtrar
1.
Phytochemistry ; 169: 112162, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31627115

RESUMO

Twelve undescribed triterpenoid pentacyclic glycosides, medicagenic acid (3-O-ß-D-glucuronopyranosyl-28-O-{[ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)]-[α-L-rhamnopyranosyl-(1 → 3)]-4-O-acetyl-ß-D-fucopyranosyl-(1→)}-2ß,3ß-dihydroxyolean-12-ene-23,28-dioic acid, 3-O-ß-D-glucuronopyranosyl-28-O-{[α-L-rhamnopyranosyl-(1 → 2)]-[ß-D-apiofuranosyl-(1 → 3)]-4-O-acetyl-ß-D-fucopyranosyl-(1→)}-2ß,3ß-dihydroxyolean-12-ene-23,28-dioic acid, 3-O-ß-D-glucuronopyranosyl-28-O-{[α-L-rhamnopyranosyl-(1 → 2)]-3,4-O-diacetyl-ß-D-fucopyranosyl-(1→)}-2ß,3ß-dihydroxyolean-12-ene-23,28-dioic acid, 28-O-{[6-O-acetyl-ß-D-glucopyranosyl-(1 → 2)]-[2-O-acetyl-α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 6)]-ß-D-glucopyranosyl-(1→)}-2ß,3ß-dihydroxyolean-12-ene-23,28-dioic acid, 28-O-{[6-O-acetyl-ß-D-glucopyranosyl-(1 → 2)]-[3-O-acetyl-α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 6)]-ß-D-glucopyranosyl-(1→)}-2ß,3ß-dihydroxyolean-12-ene-23,28-dioic acid, 28-O-{[6-O-acetyl-ß-D-glucopyranosyl-(1 → 2)]-[4-O-acetyl-α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 6)]-ß-D-glucopyranosyl-(1→)}-2ß,3ß-dihydroxyolean-12-ene-23,28-dioic acid, 28-O-{[6-O-acetyl-ß-D-glucopyranosyl-(1 → 2)]-[ß-D-glucopyranosyl-(1 → 6)]-ß-D-glucopyranosyl-(1→)}-2ß,3ß-dihydroxyolean-12-ene-23,28-dioic acid, 28-O-{[ß-D-glucopyranosyl-(1 → 2)]-[ß-D-glucopyranosyl-(1 → 6)]-ß-D-glucopyranosyl-(1→)}-2ß,3ß-dihydroxyolean-12-ene-23,28-dioic acid), zanhic acid (3-O-ß-D-glucuronopyranosyl-28-O-{[ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)]-[α-L-rhamnopyranosyl-(1 → 3)]-4-O-acetyl-ß-D-fucopyranosyl-(1→)}2ß,3ß,16α-trihydroxyolean-12-ene-23,28-dioic acid, 3-O-ß-D-glucuronopyranosyl-28-O-{[ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)]-ß-D-fucopyranosyl-(1→)}-2ß,3ß,16α-trihydroxyolean-12-ene-23,28-dioic acid), 29-hydroxy-medicagenic acid (3-O-ß-D-glucuronopyranosyl-28-O-{[ß-D-glucopyranosyl-(1 → 3)-α-L-rhamnopyranosyl-(1 → 2)]-[α-L-rhamnopyranosyl-(1 → 3)]-4-O-acetyl-ß-D-fucopyranosyl-(1→)}-2ß,3ß,29ß-trihydroxyolean-12-ene-23,28-dioic acid) and herniaric acid (28-O-{[6-O-acetyl-ß-D-glucopyranosyl-(1 → 2)]-[α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranosyl-(1 → 6)]-ß-D-glucopyranosyl-(1→)}-2ß,3ß-dihydroxyolean-18-ene-23,28-dioic acid) were isolated from the whole plant extract of Herniaria glabra L. (Caryophyllaceae), wild growing in the Ukraine. In addition, five known triterpenoid saponins; i.e. herniariasaponins 1, 4, 5, 6, and 7 were also isolated. Their structures were elucidated by HRESIMS, 1D and 2D NMR spectroscopy, as well as by comparison with the literature data. Twelve herniariasaponins, the purified crude extract, and the saponin fraction were evaluated in vitro for their xanthine oxidase, collagenase, elastase, and tyrosinase inhibitory activity. Moreover, herniariasaponins 4, 5, and 7 were screened for their cholinesterase inhibitory potential. As a result, no or low inhibition towards the mentioned enzymes was observed.


Assuntos
Caryophyllaceae/química , Inibidores Enzimáticos/farmacologia , Extratos Vegetais/farmacologia , Saponinas/farmacologia , Animais , Colinesterases/metabolismo , Colagenases/metabolismo , Inibidores Enzimáticos/química , Inibidores Enzimáticos/isolamento & purificação , Conformação Molecular , Monofenol Mono-Oxigenase/antagonistas & inibidores , Monofenol Mono-Oxigenase/metabolismo , Elastase Pancreática/antagonistas & inibidores , Elastase Pancreática/metabolismo , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Saponinas/química , Saponinas/isolamento & purificação , Estereoisomerismo , Ucrânia , Xantina Oxidase/antagonistas & inibidores , Xantina Oxidase/metabolismo
2.
Int J Nanomedicine ; 14: 6539-6553, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31496699

RESUMO

Aim: This paper reports on the incorporation of oleic acid (OA) within nanostructured lipid carriers (OA-NLC) to improve the anti-inflammatory effects in the presence of albumin. Materials and methods: NLCs produced via hot high-shear homogenization/ultrasonication were characterized in terms of particle size, zeta potential, and toxicity. We examined the effects of OA-NLC on neutrophil activities. Dermatologic therapeutic potential was also elucidated by using a murine model of leukotriene B4-induced skin inflammation. Results: In the presence of albumin, OA-NLC but not free OA inhibited superoxide generation and elastase release. Topical administration of OA-NLC alleviated neutrophil infiltration and severity of skin inflammation. Conclusion: OA incorporated within NLC can overcome the interference of albumin, which would undermine the anti-inflammatory effects of OA. OA-NLC has potential therapeutic effects in topical ointments.


Assuntos
Portadores de Fármacos/química , Inflamação/patologia , Lipídeos/química , Nanoestruturas/química , Neutrófilos/fisiologia , Ácido Oleico/química , Soroalbumina Bovina/farmacologia , Pele/patologia , Administração Tópica , Adulto , Animais , Cálcio/metabolismo , Bovinos , Morte Celular/efeitos dos fármacos , Portadores de Fármacos/administração & dosagem , Humanos , Leucotrieno B4 , Lipídeos/toxicidade , Masculino , Camundongos Endogâmicos BALB C , Nanoestruturas/administração & dosagem , Nanoestruturas/toxicidade , Nanoestruturas/ultraestrutura , Ativação de Neutrófilo/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Ácido Oleico/toxicidade , Elastase Pancreática/metabolismo , Superóxidos/metabolismo , Adulto Jovem
3.
Mar Drugs ; 17(8)2019 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-31394844

RESUMO

Two new capnosane-based diterpenoids, flaccidenol A (1) and 7-epi-pavidolide D (2), two new cembranoids, flaccidodioxide (3) and flaccidodiol (4), and three known compounds 5 to 7 were characterized from the marine soft coral Klyxum flaccidum, collected off the coast of the island of Pratas. The structures of the new compounds were determined by extensive spectroscopic analyses, including 1D and 2D nuclear magnetic resonance (NMR) spectroscopy, and spectroscopic data comparison with related structures. The rare capnosane diterpenoids were isolated herein from the genus Klyxum for the first time. The cytotoxicity of compounds 1 to 7 against the proliferation of a limited panel of cancer cell lines was assayed. The isolated diterpenoids also exhibited anti-inflammatory activity through suppression of superoxide anion generation and elastase release in the N-formyl-methionyl-leucyl-phenylalanine/cytochalasin B (fMLF/CB)-stimulated human neutrophils. Furthermore, 1 and 7 also exhibited cytotoxicity toward the tested cancer cells, and 7 could effectively inhibit elastase release. It is worth noting that the biological activities of 7 are reported for the first time in this paper.


Assuntos
Antozoários/química , Fatores Biológicos/farmacologia , Diterpenos/farmacologia , Animais , Anti-Inflamatórios/farmacologia , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Citocalasina B/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Humanos , Espectroscopia de Ressonância Magnética/métodos , N-Formilmetionina Leucil-Fenilalanina/metabolismo , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Elastase Pancreática/metabolismo , Superóxidos/metabolismo
4.
Diabetes Res Clin Pract ; 156: 107822, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31446113

RESUMO

AIMS: To determine the prevalence of low faecal elastase-1 (FE-1) (≤200 µg/g) in type 2 diabetes (T2DM), and to test the hypothesis that pancreatic enzyme replacement therapy (PERT) would reduce postprandial glycaemia after a high-fat, high-carbohydrate meal in T2DM subjects with low FE-1. METHODS: Of 109 community-based patients who submitted stool samples, 10 had low FE-1 and 8 were recruited (6 male, 2 female, 67.8 ±â€¯3.0 years). Participants were given a high-fat, high-carbohydrate meal (718 kcal) with either pancrelipase (50,000 units) or placebo in a randomised, double-blind, crossover fashion. The primary outcome was the difference in postprandial glycaemia following PERT vs placebo, as evaluated by the incremental area under the postprandial plasma glucose curve (iAUC). Secondary outcomes included differences in gastric half-emptying time (T50) measured using scintigraphy, and C-peptide iAUC. RESULTS: The prevalence of low FE-1 in T2DM was 9.2% (95% CI 3.8-14.6%). There was no difference in postprandial glycaemia iAUC (P = 0.38), gastric emptying T50 (P = 0.69) or C-peptide iAUC (P = 0.25) after PERT compared to placebo. CONCLUSIONS: Decreased FE-1 has a relatively low prevalence in community-based patients with T2DM, and PERT does not reduce postprandial glycaemia in these patients. CLINICAL TRIAL REGISTRATION NUMBER: ACTRN12617000349347.


Assuntos
Diabetes Mellitus Tipo 2/fisiopatologia , Elastase Pancreática/metabolismo , Idoso , Fezes , Feminino , Humanos , Masculino , Prevalência
5.
Arch Pharm (Weinheim) ; 352(8): e1900061, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31338866

RESUMO

Elastase is the only enzyme that has the capability to degrade elastin and collagen, the two proteins essential for skin and bones. The synthesis of some densely substituted piperidines functionalized with the trifluoromethyl group (4a-j) was carried out. The newly prepared compounds were subjected to elastase enzyme inhibitory potential and antioxidant activity assays. Among the series, 4i (IC50 = 0.341 ± 0.001 µM) exhibited the maximum inhibition against elastase. Binding analysis delineated that the fluorine atom of ligand 4i showed hydrogen and hydrophobic bonds with Thr41 and Thr96, with bond distances of 3.84 and 5.631 Å, respectively. The obtained results indicate that these trifluoromethyl functionalized piperidine derivatives could be considered as potential candidates to treat skin disorders.


Assuntos
Hidrocarbonetos Fluorados/farmacologia , Elastase Pancreática/antagonistas & inibidores , Piperidinas/farmacologia , Inibidores de Serino Proteinase/farmacologia , Animais , Relação Dose-Resposta a Droga , Hidrocarbonetos Fluorados/síntese química , Hidrocarbonetos Fluorados/química , Ligantes , Modelos Moleculares , Estrutura Molecular , Pâncreas/enzimologia , Elastase Pancreática/metabolismo , Piperidinas/síntese química , Piperidinas/química , Inibidores de Serino Proteinase/síntese química , Inibidores de Serino Proteinase/química , Relação Estrutura-Atividade , Suínos
6.
Microb Pathog ; 134: 103595, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31201902

RESUMO

The persistence of pathogenic bacteria in the marine environment has been thoroughly investigated. The potential threat that these microorganisms pose to public health in recreational waters has always been a concern. In this study, the persistence and the response of Pseudomonas aeruginosa ATCC27853 to starvation and osmotic stress were studied after its incubation in sterilized seawater during 12 months. Three different colonial variants were isolated: A7 after one month, and A81 and A82 after 8 months of incubation period. The incubation effect on the bacterial phenotype and genotype were studied by analyzing modifications in morphology, antibiotic and metal resistance, molecular typing (PFGE and MLST), pigment production and virulence factors. The starved variants showed three different colony forms, but an indistinguishable PFGE pattern and belonged to ST155, as P. aeruginosa ATCC27853. The starved variants maintained the susceptibility to the 13 tested antibiotics, with the exception of the imipenem-resistant A82 strain, which also showed a small colony variant phenotype and the highest values of tolerance to the CuSO4 + NaCl combination. Significant differences were detected in the pigment production, the elastase activity and cytotoxic potential of the starved isolates in comparison to P. aeruginosa ATCC27853. Long-term exposure to stress, such as the incubation in seawater, was shown to induce different responses in P. aeruginosa, including virulent and resistant phenotypes.


Assuntos
Pseudomonas aeruginosa/crescimento & desenvolvimento , Pseudomonas aeruginosa/fisiologia , Água do Mar/microbiologia , Inanição , Estresse Fisiológico/fisiologia , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Contagem de Colônia Microbiana , Sulfato de Cobre/farmacologia , Farmacorresistência Bacteriana/genética , Farmacorresistência Bacteriana/fisiologia , Eletroforese em Gel de Campo Pulsado , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Variação Genética/efeitos dos fármacos , Variação Genética/fisiologia , Genótipo , Imipenem/farmacologia , Testes de Sensibilidade Microbiana , Viabilidade Microbiana , Tipagem de Sequências Multilocus , Compostos Orgânicos/metabolismo , Pressão Osmótica , Elastase Pancreática/metabolismo , Fenazinas/metabolismo , Fenótipo , Pigmentos Biológicos/metabolismo , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Piocianina/metabolismo , Tolerância ao Sal/genética , Tolerância ao Sal/fisiologia , Alinhamento de Sequência , Cloreto de Sódio/farmacologia , Estresse Fisiológico/genética , Fatores de Tempo , Fatores de Virulência
7.
Elife ; 82019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-31159921

RESUMO

Reef-building corals depend on intracellular dinoflagellate symbionts that provide nutrients. Besides sugars, the transfer of sterols is essential for corals and other sterol-auxotrophic cnidarians. Sterols are important cell components, and variants of the conserved Niemann-Pick Type C2 (NPC2) sterol transporter are vastly up-regulated in symbiotic cnidarians. Types and proportions of transferred sterols and the mechanism of their transfer, however, remain unknown. Using different pairings of symbiont strains with lines of Aiptasia anemones or Acropora corals, we observe both symbiont- and host-driven patterns of sterol transfer, revealing plasticity of sterol use and functional substitution. We propose that sterol transfer is mediated by the symbiosis-specific, non-canonical NPC2 proteins, which gradually accumulate in the symbiosome. Our data suggest that non-canonical NPCs are adapted to the symbiosome environment, including low pH, and play an important role in allowing corals to dominate nutrient-poor shallow tropical seas worldwide.


Assuntos
Antozoários/genética , Proteínas de Transporte/genética , Elastase Pancreática/genética , Esteróis/metabolismo , Simbiose/genética , Animais , Antozoários/metabolismo , Proteínas de Transporte/metabolismo , Colesterol/genética , Colesterol/metabolismo , Recifes de Corais , Dinoflagelados/genética , Dinoflagelados/metabolismo , Perfilação da Expressão Gênica , Humanos , Proteínas de Membrana/genética , Elastase Pancreática/metabolismo , Anêmonas-do-Mar/genética , Anêmonas-do-Mar/metabolismo
8.
World J Gastroenterol ; 25(22): 2699-2705, 2019 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-31235993

RESUMO

The exocrine structure is significantly affected by diabetes because of endocrine structure-function disorder within the pancreas. Exocrine pancreatic dysfunction (EPD) is the general name of the malabsorption process resulting from inadequate production, release, decreased activation, and/or insufficient degradation of enzymes required for digestion from pancreatic acinar cells. It is important to diagnose patients early and correctly, since there may be both macro- and micro-nutrient deficiency in EPD. In this paper, EPD, the diabetes-EPD relationship, and the predictive, effective factors affecting the emergence of EPD are briefly explained and summarized with contemporary literature and our experienced based on clinical, lab, and radiological findings.


Assuntos
Diabetes Mellitus Tipo 1/complicações , Diabetes Mellitus Tipo 2/complicações , Insuficiência Pancreática Exócrina/etiologia , Proteínas de Transporte/análise , Proteínas de Transporte/metabolismo , Diabetes Mellitus Tipo 1/fisiopatologia , Diabetes Mellitus Tipo 2/fisiopatologia , Terapia de Reposição de Enzimas , Insuficiência Pancreática Exócrina/diagnóstico , Insuficiência Pancreática Exócrina/terapia , Fezes/enzimologia , Humanos , Pâncreas Exócrino/enzimologia , Pâncreas Exócrino/fisiopatologia , Elastase Pancreática/análise , Elastase Pancreática/metabolismo , Inibidores da Bomba de Prótons/uso terapêutico
9.
J Enzyme Inhib Med Chem ; 34(1): 927-936, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31039625

RESUMO

Skin ageing results from enhanced activation of intracellular enzymes such as collagenases, elastases and tyrosinase, stimulated by intrinsic ageing and photoageing factors. Recently, caffeine-based cosmetics are introduced that demonstrates to slow down skin photoageing process. However, no attempts have been done so for to understand caffeine functional inhibitory activity against photoageing related enzymes. Hence, this study established the caffeine molecular interaction and inhibition activity profiles against respective enzymes using in silico and in vitro methods, respectively. Results from in silico study indicates that caffeine has comparatively good affinity with collagenase (-4.6 kcal/mol), elastase (-3.36 kcal/mol) and tyrosinase (-2.86 kcal/mol) and formed the stable protein-ligand complex as validated by molecular dynamics simulation (protein-ligand contacts, RMSD, RMSF and secondary structure changes analysis). Moreover, in vitro data showed that caffeine (1000 µg/mL) has statistically significant maximum inhibition activity of 41.86, 36.44 and 13.72% for collagenase, elastase and tyrosinase, respectively.


Assuntos
Cafeína/farmacologia , Colagenases/metabolismo , Simulação por Computador , Inibidores Enzimáticos/farmacologia , Monofenol Mono-Oxigenase/antagonistas & inibidores , Elastase Pancreática/antagonistas & inibidores , Agaricus/enzimologia , Animais , Cafeína/química , Clostridium histolyticum/enzimologia , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Técnicas In Vitro , Ligantes , Simulação de Dinâmica Molecular , Monofenol Mono-Oxigenase/metabolismo , Pâncreas/enzimologia , Elastase Pancreática/metabolismo , Relação Estrutura-Atividade , Suínos
10.
Int J Biol Macromol ; 134: 216-222, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31059739

RESUMO

The increasing use of nanoparticles in various industries has triggered the need to study their interconnection with biological macromolecules. The goal of this study was to survey the SiO2-nanoparticles efficacy on the thermal denaturation, conformation and activity of elastase at three temperatures of 303, 313 and 323K in the Tris buffer at pH of 8.5. In this work, distinct techniques such as UV-vis Spectrophotometry, Spectrofluorometry, and circular dichroism were employed. The fluorescence studies showed that SiO2 nanoparticles extremely reduced the intensity of elastase with the static mechanism. Thermodynamic parameter analysis also indicated that the process of binding of SiO2-nanoparticles to elastase was spontaneous, thereby suggesting that van der Waals forces or hydrogen bonding interactions played a key role in determining the complex stability. Far-UV circular dichroism studies further revealed that SiO2 nanoparticles could cause 9.79% reduction in the content of the α-helix and 3.24% increase in the content of the ß-sheet. Furthermore, kinetic parameters (Vmax and Kcat/Km) indicated that SiO2 nanoparticles had an activation effect on the elastase activity. Melting temperature studies at the selected concentration of SiO2 nanoparticles also showed that by adding SiO2 nanoparticles, elastase thermal stability was slightly increased. Overall, these nanoparticles modified the structure of the elastase, ultimately changing the activity and stability of this enzyme.


Assuntos
Nanopartículas , Elastase Pancreática/metabolismo , Dióxido de Silício/química , Dióxido de Silício/metabolismo , Animais , Estabilidade Enzimática , Cinética , Ligação Proteica , Análise Espectral , Suínos , Temperatura
11.
J Microencapsul ; 36(2): 180-191, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31070486

RESUMO

The study was aimed to evaluate the effectiveness of rosmarinic acid (RA) loaded ethosomes (ETHs) and liposomes (LPs) when subjected to the transdermal application. RA-loaded ETHs and LPs were prepared, optimised, and characterised. The ex vivo permeation studies of formulations using mouse abdominal skin were performed. Antioxidant activities and the inhibitory effects of formulations on collagenase and elastase enzymes were measured. Optimised ethosomal formulation (F3) was showed nanometric size range (138 ± 1.11 nm) and greatest entrapment (55 ± 1.80%), was selected for further transdermal permeation studies. Skin permeation profile of the nanoformulations analysed by HPLC revealed an enhanced permeation of ETHs. Transdermal flux of ETHs was found to be higher than RA solution and LPs. Enzyme inhibitions of ETHs were the significant difference found between ETHs and LPs (p < 0.05). ETHs were found to be more effective and successful than LPs. Results suggest that ETHs are more effective than LPs for transdermal delivery of RA.


Assuntos
Antioxidantes/administração & dosagem , Cinamatos/administração & dosagem , Depsídeos/administração & dosagem , Portadores de Fármacos/química , Inibidores de Metaloproteinases de Matriz/administração & dosagem , Administração Cutânea , Animais , Antioxidantes/farmacocinética , Antioxidantes/farmacologia , Linhagem Celular , Cinamatos/farmacocinética , Cinamatos/farmacologia , Colagenases/metabolismo , Depsídeos/farmacocinética , Depsídeos/farmacologia , Lipossomos/química , Inibidores de Metaloproteinases de Matriz/farmacocinética , Inibidores de Metaloproteinases de Matriz/farmacologia , Camundongos , Elastase Pancreática/antagonistas & inibidores , Elastase Pancreática/metabolismo , Pele/efeitos dos fármacos , Pele/metabolismo , Absorção Cutânea , Envelhecimento da Pele/efeitos dos fármacos
12.
Bioengineered ; 10(1): 87-97, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-30957636

RESUMO

Expression of recombinant proteins fused to a novel glycomodule tag, termed hydroxyproline (Hyp)-O-glycosylated peptides (HypGP), was earlier found to boost secreted protein yields up to 500-fold in plant cell culture. Here, this technology was applied to the expression of human protease inhibitor α1-antitrypsin (AAT) in tobacco BY-2 cell culture. A designer HypGP tag composed of a 'Ala-Pro' motif of 20 units, or (AP)20, was engineered either at the N- or C-terminal end of AAT. The (AP)20 tag substantially increased the secreted yields of the recombinant AAT up to 34.7 mg/L. However, the (AP)20-tagged AAT products were frequently subjected to proteolytic processing. The intact AAT-(AP)20 along with some of the truncated AAT domains exhibited desired biological activity in inhibiting elastase. The results from this research demonstrated that the designer (AP)20 module engineered in BY-2 cells could function as a molecular carrier to substantially enhance the secreted yields of the recombinant AAT.


Assuntos
Elastase Pancreática/antagonistas & inibidores , Processamento de Proteína Pós-Traducional , Proteínas Secretadas Inibidoras de Proteinases/biossíntese , Proteínas Recombinantes de Fusão/biossíntese , Tabaco/genética , alfa 1-Antitripsina/biossíntese , Sequência de Bases , Técnicas de Cultura de Células , Dipeptídeos/genética , Dipeptídeos/metabolismo , Expressão Gênica , Glicosilação , Humanos , Elastase Pancreática/metabolismo , Peptídeos/genética , Peptídeos/metabolismo , Células Vegetais/metabolismo , Plasmídeos/química , Plasmídeos/metabolismo , Proteínas Secretadas Inibidoras de Proteinases/genética , Proteínas Secretadas Inibidoras de Proteinases/isolamento & purificação , Proteínas Secretadas Inibidoras de Proteinases/farmacologia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/farmacologia , Tabaco/citologia , Tabaco/metabolismo , Transformação Genética , alfa 1-Antitripsina/genética , alfa 1-Antitripsina/isolamento & purificação , alfa 1-Antitripsina/farmacologia
14.
Photochem Photobiol Sci ; 18(5): 1259-1274, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30891584

RESUMO

Topical delivery of potent antioxidants maintain the redox balance of the skin, which leads to the downregulation of matrix metalloproteinase (MMP) expression and prevents UV radiation-induced photoaging. In this study, we aimed at investigating the inhibitory role of silk cocoon extract (SCE) isolated from the Antheraea assamensis (AA), Bombyx mori (BM), and Philosamia ricini (PR) silk varieties against UV radiation-induced MMP expression. Incubation of elastase and hyaluronidase with Antheraea assamensis silk cocoon extract (AASCE) caused 50% inhibition of activity. The assessment of total collagen content using the Sirius red assay showed that AASCE (10 µg mL-1) and Philosamia ricini silk cocoon extract (PRSCE at 100 µg mL-1 concentration) post-treatment significantly enhanced the total collagen content in UVA1 and UVB irradiated HDF cells, whereas BM silk cocoon extract (BMSCE at 100 µg mL-1 concentration) post-treatment significantly enhanced the total collagen content in UVA1-irradiated HDF cells. Gene expression studies revealed AASCE and PRSCE post-treatment downregulated the expression of interleukin (IL)-6, MMP-1 and upregulated procollagen genes in UV irradiated HDF cells. Gelatin zymography studies with AASCE post-treatment downregulated the release of MMP-2 and MMP-9 by HaCaT cells. The overall results validate AASCE efficiently shielding UV radiation-induced collagen and elastin degradation by downregulation of MMP expression, substantiating its further use as a potent antioxidant complement in skin care formulations.


Assuntos
Colágeno/farmacologia , Inibidores Enzimáticos/farmacologia , Hialuronoglucosaminidase/antagonistas & inibidores , Metaloproteinases da Matriz/biossíntese , Elastase Pancreática/antagonistas & inibidores , Seda/química , Animais , Bombyx , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Colágeno/química , Colágeno/isolamento & purificação , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/química , Inibidores Enzimáticos/isolamento & purificação , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Hialuronoglucosaminidase/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Elastase Pancreática/metabolismo , Espécies Reativas de Oxigênio/análise , Espécies Reativas de Oxigênio/metabolismo , Seda/metabolismo , Relação Estrutura-Atividade , Raios Ultravioleta
15.
Equine Vet J ; 51(5): 641-645, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30849189

RESUMO

BACKGROUND: Inhaled corticosteroids are effective in the treatment of equine asthma. A recent study reported that nebulisation of injectable dexamethasone had low systemic bioavailability in healthy horses and could represent a cost-effective therapy for equine inflammatory lung diseases. OBJECTIVES: To determine the effects of dexamethasone nebulisation on lung function in severe asthmatic horses. It was hypothesised that dexamethasone administered by nebulisation would be more effective than the same dose administered orally. STUDY DESIGN: Randomised blinded experimental study in severe asthmatic horses. METHODS: Twelve severe asthmatic horses in clinical exacerbation were randomly assigned to treatment with 5 mg of dexamethasone sodium phosphate by nebulisation (n = 6) or by oral administration (n = 6), once daily for 7 days. Lung function was evaluated at baseline, after four treatment days (D4) and 24 h after the last dose (D8). The presence of residual bronchospasm was assessed on D8 with N-butylscopolammonium bromide administration (0.3 mg/kg i.v.). A respiratory clinical score was performed daily. Serum cortisol concentration was measured at baseline, D4 and D8. RESULTS: The pulmonary elastance was unchanged in both groups while pulmonary resistance was significantly improved in the oral group on D8 (mean reduction in 1 cm H2 O/L/s [CI: 0.34-1.65, P = 0.003]). All horses had residual bronchospasm at the end of the study. There was a group difference in the respiratory clinical score as it was significantly reduced in the oral group, from D5 to D8. Serum cortisol concentration decreased in all subjects. MAIN LIMITATIONS: Low number of horses and lack of placebo group. CONCLUSIONS: Considering the lack of improvement of lung function and the hypothalamic-pituitary-adrenal axis suppression, the results of this study do not support aerosolisation of an injectable form of dexamethasone for the treatment of severe equine asthma at the dose and with the nebuliser evaluated.


Assuntos
Asma/veterinária , Dexametasona/análogos & derivados , Doenças dos Cavalos/tratamento farmacológico , Administração por Inalação , Corticosteroides/uso terapêutico , Aerossóis , Animais , Dexametasona/administração & dosagem , Dexametasona/uso terapêutico , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Cavalos , Pulmão/enzimologia , Pulmão/metabolismo , Elastase Pancreática/metabolismo
16.
Biofabrication ; 11(3): 035012, 2019 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-30921774

RESUMO

Decellularized tissue matrices are promising substrates for tissue generation by stem cells to replace poorly regenerating tissues such as cartilage. However, the dense matrix of decellularized cartilage impedes colonisation by stem cells. Here, we show that digestion of elastin fibre bundles traversing auricular cartilage creates channels through which cells can migrate into the matrix. Human chondrocytes and bone marrow-derived mesenchymal stromal cells efficiently colonise elastin-treated scaffolds through these channels, restoring a glycosaminoglycan-rich matrix and improving mechanical properties while maintaining size and shape of the restored tissue. The scaffolds are also rapidly colonised by endogenous cartilage-forming cells in a subcutaneously implanted osteochondral biopsy model. Creating channels for cells in tissue matrices may be a broadly applicable strategy for recellularization and restoration of tissue function.


Assuntos
Cartilagem da Orelha/citologia , Elastase Pancreática/metabolismo , Adolescente , Idoso , Animais , Bovinos , Criança , Condrogênese , Elastina/metabolismo , Matriz Extracelular/química , Feminino , Glicosaminoglicanos/metabolismo , Humanos , Camundongos Nus , Pessoa de Meia-Idade , Tecidos Suporte/química
17.
Artigo em Inglês | MEDLINE | ID: mdl-30873390

RESUMO

Background: Pseudomonas aeruginosa causes severe chronic respiratory diseases and is associated with recalcitrant chronic rhinosinusitis (CRS). P. aeruginosa exoproteins contain virulence factors and play important roles in the pathogenicity of P. aeruginosa, however their role in CRS pathophysiology remains unknown. Methods: We isolated P. aeruginosa clinical isolates (CIs) and obtained clinical information from 21 CRS patients. Elastase activity of the CIs was measured at different phases of growth. Primary human nasal epithelial cells (HNECs) were cultured at air-liquid interface (ALI) and challenged with P. aeruginosa exoproteins or purified elastase, followed by measuring Transepithelial Electrical Resistance (TEER), permeability of FITC-dextrans, western blot, and immunofluorescence. Results: 14/21 CIs had a significant increase in elastase activity in stationary phase of growth. There was a highly significant strong correlation between the in vitro elastase activity of P. aeruginosa CIs with mucosal barrier disruption evidenced by increased permeability of FITC-dextrans (r = 0.95, p = 0.0004) and decreased TEER (r = -0.9333, P < 0.01) after 4 h of challenge. Western blot showed a significant degradation of ZO-1, Occludin and ß-actin in relation to the elastase activity of the exoproteins. There was a highly significant correlation between the in vitro elastase activity of P. aeruginosa CIs and CRS disease severity (for log phase, r = 0.5631, p = 0.0097; for stationary phase, r = 0.66, p = 0.0013) assessed by CT imaging of the paranasal sinuses. Conclusion: Our results implicate P. aeruginosa exoproteins as playing a major role in the pathophysiology of P. aeruginosa associated CRS by severely compromising mucosal barrier structure and function.


Assuntos
Células Epiteliais/microbiologia , Elastase Pancreática/metabolismo , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/metabolismo , Sinusite/microbiologia , Fatores de Virulência/metabolismo , Adulto , Idoso , Células Cultivadas , Doença Crônica , Células Epiteliais/fisiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infecções por Pseudomonas/fisiopatologia , Pseudomonas aeruginosa/isolamento & purificação , Sinusite/fisiopatologia
18.
Int J Biol Macromol ; 131: 473-483, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30880056

RESUMO

In this study, the impact of spermidine on the stability, conformation and activity of elastase was investigated at the pH of 8.5 (the optimum pH for elastase) and different temperatures (303, 313, and 323 K) using UV-vis spectrophotometry, Spectrofluorimetry, circular dichroism, and enzyme activity measurements. The empirical results were obtained and compared with those achieved by the molecular docking simulation. Spectrofluorometric results proved that with the addition of spermidine to the protein solution, the emission severity of elastase was extremely reduced. Further, the Stern-Volmer equation demonstrated that quenching was principally of the static type. Moreover, ∆H0 and ∆S0 showed a negative value, revealing that hydrogen bonds or van der Waals interactions played a critical role in the interaction between spermidine and elastase. Km and kcat [E] parameters also showed that spermidine acted as an activator for elastase. Far-UV circular dichroism also revealed that spermidine could alter the secondary structure of elastase via a partial increment within the content of the α-helix structure (from 7.8 to 8.6), while it was somewhat diminished in the ß-sheet (from 29.4 to 28.8). Molecular docking simulation results also demonstrated that spermidine could bind to porcine elastase, and van der Waals forces or hydrogen bonding interactions played the main role in this binding. Spermidine, therefore, served as a partial stabilizer and an activator for the enzyme.


Assuntos
Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Elastase Pancreática/química , Análise Espectral , Espermidina/química , Sítios de Ligação , Elastase Pancreática/metabolismo , Ligação Proteica , Conformação Proteica , Estabilidade Proteica , Espermidina/metabolismo , Termodinâmica
19.
Mar Drugs ; 17(2)2019 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-30781569

RESUMO

Three new eunicellin-derived diterpenoids of briarellin type, briarenones A‒C (1‒3), were isolated from a Formosan gorgonian Briareum violaceum. The chemical structures of the compounds were elucidated on the basis of extensive spectroscopic analyses, including two-dimensional (2D) NMR. The absolute configuration of 1 was further confirmed by a single crystal X-ray diffraction analysis. The in vitro cytotoxic and anti-inflammatory potentialities of the isolated metabolites were tested against the growth of a limited panel of cancer cell lines and against the production of superoxide anions and elastase release in N-formyl-methionyl-leucyl-phenyl-alanine and cytochalasin B (fMLF/CB)-stimulated human neutrophils, respectively.


Assuntos
Cnidários/química , Diterpenos/química , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Citocalasinas/farmacologia , Diterpenos/isolamento & purificação , Humanos , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Conformação Molecular , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Elastase Pancreática/metabolismo , Superóxidos/metabolismo , Difração de Raios X
20.
Immunol Lett ; 207: 36-45, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30738096

RESUMO

Resolution of inflammation needs effective and timely removal of dead cells and other toxic products of neutrophils, monocytes, and macrophages. In this study, we evaluated the role of monocytes in the clearance of neutrophil extracellular trap (NET) and apoptotic neutrophils in the inflammation site. For this, monocytes were observed microscopically after exposing them with NETs and/or apoptotic bodies. A subset of monocytes exposed to NETs ejected extracellular traps and this was shown to be mediated by proteins like elastase and citrullinated histones present in NET supernatant. Monocytes showed a preference for the internalisation of the apoptotic body when both NET and apoptotic bodies were present in the medium. The study provides new insight into the role of monocytes in the clearance of NET and apoptotic neutrophils and this information may open up a way in formulating therapeutic strategies for accelerating resolution of inflammation.


Assuntos
Armadilhas Extracelulares/metabolismo , Histonas/metabolismo , Inflamação/imunologia , Macrófagos/imunologia , Monócitos/imunologia , Neutrófilos/imunologia , Elastase Pancreática/metabolismo , Adulto , Apoptose , Células Cultivadas , Citrulinação , Vesículas Extracelulares/metabolismo , Feminino , Voluntários Saudáveis , Humanos , Masculino , Fagocitose , Adulto Jovem
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA