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1.
Rev. Ciênc. Méd. Biol. (Impr.) ; 20(4): 510-519, fev 11, 2022. tab
Article in Portuguese | LILACS | ID: biblio-1359304

ABSTRACT

Introdução: a oxidação em sistemas biológicos está relacionada ao desenvolvimento de patologias em humanos. A ingestão de alimentos ricos em compostos químicos que exercem atividade antioxidante contribui para a prevenção e redução dos efeitos deletérios dos radicais livres formados no organismo. Peptídeos derivados das caseínas têm mostrado um elevado potencial como agentes antioxidantes. Objetivos: neste sentido, o presente estudo avaliou a atividade antioxidante de hidrolisados derivados de caseínas de leites das espécies bubalina, bovina e caprina, obtidos pela ação de diferentes proteases. Metodologia: inicialmente, as caseínas foram isoladas dos demais componentes do leite, depois foram submetidas ao processo de proteólise pelas enzimas bromelina, papaína, tripsina e neutrase, individualmente. A atividade antioxidante dos hidrolisados foi avaliada, através da capacidade de eliminação dos radicais: hidroxila (OH­Ë™), superóxido (O2­Ë™), 2,2 difenil-1-picrilhidrazil (DPPH˙), 2,2'azinobis-(3-ácido etilbenzotiazolino-6-sulfônico (ABTS˙), e quelante dos íons metálicos cobre (Cu2+) e ferro (Fe2+). Resultados: os resultados mostraram que a caseína bovina apresentou o menor (35,54%) grau de hidrólise e a caseína bubalina apresentou o maior (85,64%) grau de hidrólise pela ação da neutrase e bromelina após 480 minutos, respectivamente. O potencial para o sequestro dos radicais hidroxila variou entre 0 e 100%, superóxido superior a 80%, ABTS superior a 85%, DPPH entre 20 e 95% habilidade de quelar ferro entre 10 e 100% e cobre entre 14 e 80%. Conclusão: assim, a hidrólise das caseínas do leite bubalino, bovino e caprino foram capazes de produzir hidrolisados com elevado potencial antioxidante e que, mediante novos estudos, poderá vir ser incorporado em produtos alimentícios para o consumo humano.


Introduction: oxidation in biological systems is related to the development of pathologies in humans. The ingestion of foods rich in chemical compounds that exert antioxidant activity contributes to the prevention and reduction of the deleterious effects of free radicals formed in the body. Peptides derived from caseins have shown high potential as antioxidant agents. Objectives: the present study evaluated the antioxidant activity of casein hydrolysates derived from bubaline, bovine, and caprine milk obtained by the action of different proteases. Methodology: initially, the caseins were isolated from the other milk components, and then subjected to the proteolysis process by the enzymes bromelain, papain, trypsin and neutrase, individually. The antioxidant activity of the hydrolysates was evaluated, through the capacity of elimination of the radicals: hydroxyl (OH-˙), superoxide (O2-˙), 2,2 diphenyl-1-picrylhydrazyl (DPPH˙), 2,2'azinobis-(3-ethylbenzothiazolino-6-sulfonic acid (ABTS˙), and chelating of the metal ions copper (Cu2+) and iron (Fe2+). Results: the results showed that bovine casein showed the lowest (35.54%) degree of hydrolysis and bubaline casein showed the highest (85.64%) degree of hydrolysis by the action of neutrase and bromelin after 480 minutes, respectively. The potential for hydroxyl radical sequestration varied between 0 and 100%, superoxide higher than 80%, ABTS higher than 85%, DPPH between 20 and 95% and the ability to chelate iron between 10 and 100% and copper between 14 and 80%. Conclusion: thus, the hydrolysis of caseins from bubaline, bovine and goat milk were able to produce hydrolysates with high antioxidant potential and that, upon further studies, may be incorporated into food products for human consumption.


Subject(s)
Animals , Cattle , Peptides , Buffaloes , Cattle , Goats , Dietary Supplements
2.
Article in Chinese | WPRIM | ID: wpr-940983

ABSTRACT

OBJECTIVE@#To explore whether the using of mimetic peptide Gap27, a selective inhibitor of connexin 43 (Cx43), could block the death of dopamine neurons and influence the expression of Cx43 in 6-hydroxydopamine (6-OHDA)-induced Parkinson's disease mouse models.@*METHODS@#Eighteen C57BL/6 mice were randomly divided into control group, 6-OHDA group and 6-OHDA+Gap27 group, with 6 mice in each group. Bilateral substantia nigra stereotactic injection was performed. The control group was injected with ascorbate solution, 6-OHDA group was injected with 6-OHDA solution, and 6-OHDA+Gap27 group was injected with 6-OHDA and Gap27 mixed solution. Immuno-histochemical staining was used to detect the number of dopamine neurons, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of Cx43 messenger ribonucleic acid (mRNA), immuno-fluorescence staining was used to detect the distribution of Cx43 protein, the contents of Cx43 protein and Cx43 phosphorylation at serine 368 (Cx43-ps368) in mouse midbrain were detected by Western blot.@*RESULTS@#After injection of 6-OHDA, numerous dopamine neurons in substantia nigra died as Cx43 content increased, Cx43-ps368 content decreased. Mixing Gap27 while injecting 6-OHDA could reduce the number of death dopamine neurons and weaken the changes of Cx43 and Cx43-ps368 content caused by 6-OHDA. The number of tyrosine hydroxylase (TH) immunoreactive positive neurons in 6-OHDA group decreased to 27.7% ± 0.02% of the control group (P < 0.01); The number of TH immunoreactive positive neurons in 6-OHDA+Gap27 group was (1.64±0.16) times higher than that in 6-OHDA group (P < 0.05); The content of total Cx43 protein in 6-OHDA group was (1.44±0.07) times higher than that in 6-OHDA+Gap27 group (P < 0.05) while (1.68±0.07) times higher than that in control group (P < 0.01). In 6-OHDA group, the content of Cx43-ps368 protein and its proportion in total Cx43 protein were significantly lower than that in 6-OHDA+Gap27 group (P < 0.05).@*CONCLUSION@#In 6-OHDA mouse models, mimetic peptide Gap27 played a protective role in reducing the damage to substantia nigra dopamine neurons, which was induced by 6-OHDA. The overexpression of Cx43 protein might have neurotoxicity to dopamine neuron. Meanwhile, decreasing Cx43 protein level and keeping Cx43-ps368 protein level may be the protective mechanisms of Gap27.


Subject(s)
Animals , Connexin 43/pharmacology , Disease Models, Animal , Dopaminergic Neurons/metabolism , Mice , Mice, Inbred C57BL , Oxidopamine/metabolism , Parkinson Disease/metabolism , Peptides/pharmacology , Tyrosine 3-Monooxygenase/pharmacology
3.
Article in Chinese | WPRIM | ID: wpr-928446

ABSTRACT

OBJECTIVE@#To characterize a novel HLA allele, A*24:191, its DNA sequence, MHC modeling structure, and the possible influence of the amino-acid residue variations on the molecule.@*METHODS@#The HLA sequence was determined by Luminex PCR-SSO and PCR-SBT. Its MHC molecular structure and the possible effects of the amino-acid residue variations were modeled and analyzed with Phyre2, RCSB PDB and HistoCheck software.@*RESULTS@#The PCR-SBT revealed the novel A*24:191 differs from A*24:02 in exon 2 at position 256, 265, 270 with G>C, G>C, A>T. The MHC molecular structure prediction showed that, compared with A*24:02, the 62nd residue of A*24:191 changed from the acidic E to a neutral Q, both with the side chain extending outside the α helix pointing forward the groove, (Risler's score, R=2), the 65th changed from the smaller neutral G extending inside the helix to a basic R with a long-chain extending upward outside the helix (R=52), and the 66th changed from the basic K to a neutral N both with a long side chain extending inside the groove (R=31). The above residues are located on the α helix of the α 1 domain which constituting the side wall of the peptide-binding groove. The DSS Score=3.85. From the surface image of the molecule, it can be clearly seen that the variations of the properties, sizes and configurations of the residues caused significant changes in the shape of the surface structure of the α helix.@*CONCLUSION@#It suggested that the residue variations are likely to change the peptide binding properties as well as the TCR and antibody binding characteristics of the molecule.


Subject(s)
Alleles , Amino Acid Sequence , HLA-A Antigens , Humans , Peptides , Protein Binding , Protein Conformation
4.
Article in Chinese | WPRIM | ID: wpr-928237

ABSTRACT

This study aims to explore the potential of polyaspartic acid grafted dopamine copolymer (PAsp- g-DA) chelated Fe 3+ for magnetic resonance imaging (MRI) visual photothermal therapy. Polyaspartic acid grafted copolymer of covalently grafted dopamine and polyethylene glycol (PAsp- g-DA/PEG) was obtained by the ammonolysis reaction of poly succinimide (PSI), and then chelated with Fe 3+ in aqueous solution. The relaxivity in vitro, magnetic resonance imaging enhancement in vivo and photothermal conversion effect at 808 nm were investigated. The results showed that polymeric iron coordination had good near-infrared absorption and photothermal conversion properties, good magnetic resonance enhancement effect, and good longitudinal relaxation efficiency under different magnetic field intensities. In summary, this study provides a new magnetic resonance visual photothermal therapeutic agent and a new research idea for the research in related fields.


Subject(s)
Dopamine , Magnetic Resonance Imaging/methods , Nanoparticles , Peptides , Phototherapy , Photothermal Therapy , Polymers
5.
Article in Chinese | WPRIM | ID: wpr-928053

ABSTRACT

This paper explored the specific peptides from Bubali Cornu by ultra-performance liquid chromatography-tandem mass spectrometry and based on mathematics set theory. Following the profile analysis of peptides from Bubali Cornu, Bovis Grunniens Cornu, Caprae Hircus Cornu, and Suis Cornu by nano LC-LTQ-Obitrap-MS after digestion with trypsin, the relationship of peptide composition among different samples was analyzed using the mathematics set theory. The ones that existed only in the Bubali Cornu set rather than in any other set were considered as the specific peptides of Bubali Cornu. The further bioinformatic analysis revealed four specific peptides from Bubali Cornu, whose specificity was verified by UPLC-QQQ-MS. The results showed that these four peptides could be used for distinguishing Bubali Cornu from Caprae Hircus Cornu and Suis Cornu. This study has provided a rapid and simple method for seeking the specific peptides in animal medicines, which can be utilized for quality evaluation of animal medicines, thus making them authenticable and traceable.


Subject(s)
Animals , Chromatography, Liquid , Cornus , Horns/chemistry , Peptides/chemistry , Tandem Mass Spectrometry
6.
Chinese Journal of Biotechnology ; (12): 1209-1217, 2022.
Article in Chinese | WPRIM | ID: wpr-927775

ABSTRACT

Recombinant HLA-Ⅰ molecules/antigenic peptide complexes (pHLA complexes) are applied in the research of human T cell-specific immune responses. The preparation of pHLA complex is based on genetic engineering and protein in vitro dilution and folding-refolding technology. In an in vitro refolding system, recombinant HLA-Ⅰ molecules correctly fold and bind with antigenic peptides to form complexes. In this study, ultrafiltration-high performance liquid chromatography (ultrafiltration-HPLC) was used for quantitative determination of the antigenic peptides in recombinant pHLA complexes, especially for those in a small amount of prepared products. By adding the recombinant HLA-Ⅰ molecules and antigenic peptides into the refolding buffer, the heavy chain (HC) and light chain (β2m) of recombinant HLA-Ⅰ molecules were refolded and bond with the VYF antigenic peptide containing anchor residues to form a pHLA complex. The unbound free antigenic peptide VYF was removed by ultrafiltration to retain the complex. Finally, the pHLA complex was treated by acid to destroy its interaction, thus releasing the antigenic peptide. The results showed that the prepared recombinant pHLA complex was recognized by HLA-Ⅰ molecule specific antibody W6/32, which indicated that the recombinant HLA-Ⅰ class molecule had correct folding and was identified as pHLA complex. The antigen peptide VYF contained in the pHLA complex was also detected by ultrafiltration-HPLC, so it is feasible to apply ultrafiltration-HPLC for determination of pHLA complex. Compared with Western blotting, the concentration of antigenic peptides detected by ultrafiltration-HPLC was 0-9 μg/mL. The binding conditions can be optimized according to the amount of antigenic peptides bound in the complex in order to improve the folding efficiency of HLA-Ⅰ molecules and promote the binding of HLA-Ⅰ molecules to antigenic peptides. The production rate of pHLA complexes in the refolding system can also be calculated according to the content of antigenic peptides bound by pHLA complexes. Therefore, ultrafiltration-HPLC in this study can be used for the quality control of the preparation process of pHLA complexes, and may facilitate the research of T cell-specific immunity, artificial antigen-presenting cells, and development of specific tetramer probe applications.


Subject(s)
Amino Acid Sequence , Antigens , Chromatography, High Pressure Liquid , Humans , Peptides/chemistry , Ultrafiltration
7.
Chinese Journal of Biotechnology ; (12): 650-665, 2022.
Article in Chinese | WPRIM | ID: wpr-927734

ABSTRACT

Based on the self-assembly process occurring in the human body all the time, self-assembled nanomaterials were designed by the researchers. The self-assembled nanomaterials have controllability, biocompatibility and functional advantages in vivo. The self-assembled nanomaterials constructed in situ under a physiological environment display various biological characteristics which can be used for imaging, therapy, and broad clinical applications. In situ self-assembled nanomaterials can boost drug function, reduce toxic and side effects, prolong imaging time and enlarge signal-to-noise ratio. By using pathological conditions to trigger specific responses in vivo, well-ordered nanoaggregates can be spontaneously formed by multiple weak bonding interactions. The assembly shows higher accumulation and longer retention in situ. Endogenous triggers for in situ assembly, such as enzymes, pH, reactive oxygen species and ligand receptor interaction, can be used to transform the materials into a variety of controllable nanostructures including nanoparticles, nanofibers and gels through bioactivated in vivo assembly (BIVA) strategies. BIVA strategies can be applied for treatment, imaging or participate in the physiological activities of cells at the lesion site. This review summarized and prospected the design of self-assembled peptide materials based on BIVA technology and their biomedical applications. The nanostructures of the self-assembly enable some beneficial biological effects, such as assembly induced retention (AIR) effect, enhanced targeting effect, multivalent bond effect, and membrane disturbance. Thus, the BIVA nanotechnology is promising for efficient drug delivery, enhancement of targeting and treatment, as well as optimization of the biological distribution of drugs.


Subject(s)
Drug Delivery Systems , Humans , Nanofibers/chemistry , Nanoparticles , Nanostructures/chemistry , Peptides
8.
Chinese Journal of Biotechnology ; (12): 139-147, 2022.
Article in Chinese | WPRIM | ID: wpr-927699

ABSTRACT

The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β2m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a SuperdexTM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.


Subject(s)
Animals , Capripoxvirus , Epitopes, T-Lymphocyte/genetics , Peptides/genetics , Poxviridae Infections , Sheep , Sheep Diseases
9.
Acta cir. bras ; 37(6): e370605, 2022. graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1402959

ABSTRACT

Purpose: Traumatic brain injury (TBI) is a major cause of death and disability. Cerebrolysin (CBL) has been reported to be anti-inflammatory by reducing reactive oxygen species (ROS) production. However, the neuroprotection of CBL in TBI and the potential mechanism are unclear. We aimed to investigate the neuroprotection and mechanisms of CBL in TBI. Methods: The TBI model was established in strict accordance with the Feeney weight-drop model of focal injury. The neurological score, brain water content, neuroinflammatory cytokine levels, and neuronal damage were evaluated. The involvement of the early brain injury modulatory pathway was also investigated. Results: Following TBI, the results showed that CBL administration increased neurological scores and decreased brain edema by alleviating blood­brain barrier (BBB) permeability, upregulating tight junction protein (ZO­1) levels, and decreasing the levels of the inflammatory cytokines tumor necrosis factor­α (TNF­α), interleukin­1ß (IL­1ß), IL­6, and NF­κB. The TUNEL assay showed that CBL decreased hippocampal neuronal apoptosis after TBI and decreased the protein expression levels of caspase­3 and Bax, increasing the levels of Bcl­2. The levels of Toll­like receptor 2 (TLR2) and TLR4 were significantly decreased after CBL treatment. In TBI patients, CBL can also decrease TNF­α, IL­1ß, IL­6, and NF­κB levels. This result indicates that CBL­mediated inhibition of neuroinflammation and apoptosis ameliorated neuronal death after TBI. The neuroprotective capacity of CBL is partly dependent on the TLR signaling pathway. Conclusions: Taken together, the results of this study indicate that CBL can improve neurological outcomes and reduce neuronal death against neuroinflammation and apoptosis via the TLR signaling pathway in mice.


Subject(s)
Animals , Mice , Peptides/administration & dosage , Reactive Oxygen Species/analysis , Apoptosis , Brain Injuries, Traumatic/therapy , Neuroinflammatory Diseases/veterinary
10.
Braz. J. Pharm. Sci. (Online) ; 58: e191093, 2022. tab, graf
Article in English | LILACS | ID: biblio-1383999

ABSTRACT

Abstract In recent years, improvements have been made, through biotechnological processes, in the production and development of peptides capable of increasing collagen and elastin synthesis for anti-aging skin care. However, proteins have many limitations due to their structural, chemical and physical fragility to external aggressions, which may cause conformational changes, leading to loss of biological activity. Therefore, it is important to create delivery systems that protect these biomolecules from damage, allowing them to reach their target. This work aimed to develop a system able to carry bovine serum albumin (BSA), used as a model of a protein, and to incorporate this system in a semisolid formulation suitable for skin application. A microemulgel based on a solid-in-oil-in-water (S/O/W) microemulsion was prepared. Firstly, the association efficiency (AE) of lyophilized BSA-sucrose ester complex and the size of S/O nanodispersion were assessed; then, the characterization and stability evaluation of the final semisolid formulation through evaluation of pH, texture and rheological behavior were performed. The average value of AE was 54.74% ± 2.17. It was possible to develop an S/O/W microemulsion, which allowed the subsequent development of an S/O/W microemulgel that assured suitable pH, texture and rheological characteristics for skin application.


Subject(s)
Serum Albumin, Bovine , Proteins/adverse effects , Collagen , Peptides/agonists , Skin/drug effects , Biological Products , Aging , Hydrogen-Ion Concentration
11.
Infectio ; 25(4): 241-249, oct.-dic. 2021. tab, graf
Article in English | LILACS, COLNAL | ID: biblio-1286717

ABSTRACT

Abstract Infection through the Hepatitis C virus does not have a vaccine and treatment with pegylated interferon and ribavirin can fail; which is why it may cause chronic infection and, consequently, could develop liver failure or hepatocellular carcinoma. It has been described that virus-cell recognition occurs between the E2 viral envelope protein and diverse cell receptors, with this interaction being critical in viral infection. which is why the study sought to identify inhibitory peptides of the interaction between viral E2 protein and the CD81 and CD209 receptors. Methodology: Through the RCSB protein database, crystals from the CD81 and CD209 receptors were selected, CD81/E2-HCV, CD209/E2-HCV complexes were carried out by SWISS-MODEL to generate inhibitory peptides of protein interaction through the Rosetta web server, this interaction was validated through ClusPro and finally, determined the theoretical physicochemical and cytotoxic properties of these peptides. Results: two peptides were obtained, without predicted toxicity, with a theoretical capacity of blocking the protein interaction between the E2 protein of the virus and CD81 and CD209.


Resumen La infección por el virus de la hepatitis C, no cuenta con vacuna y el tratamiento con interferón pegilado y ribavirina puede fallar; por lo que puede causar infec ción crónica y como consecuencia podría desarrollarse falla hepática o carcinoma hepatocelular. Se ha descrito que el reconocimiento virus-célula, se da entre la proteína de envoltura viral E2 y diversos receptores celulares, siendo esta interacción crítica en la infección viral. Razón por la cual este estudio buscó identificar péptidos inhibidores de la interacción entre la proteína E2 viral y los receptores CD81 y CD209. Metodología: A través de la base de datos de proteínas RCSB, se seleccionaron cristales de los receptores CD81 y CD209, se realizaron complejos CD81/E2-HCV, CD209/E2-HCV para generar péptidos inhibidores de interacción proteica a través del servidor web Rosetta, esta interacción fue validada a través de ClusPro y finalmente se evaluaron las propiedades fisicoquímicas y citotóxicas teóricas para estos péptidos. Resultados: se obtuvo dos péptidos, sin toxicidad predicha, con capacidad teórica de bloquear la interacción proteica entre la proteína E2 del virus y CD81 y CD209.


Subject(s)
Humans , Hepatitis Viruses , Peptides , Vaccines , Proteins , Hepatitis C , Liver Failure , Hepacivirus , Infections
12.
Vitae (Medellín) ; 28(3): 1-14, 2021-08-11. Ilustraciones
Article in English | LILACS, COLNAL | ID: biblio-1363261

ABSTRACT

Background: Milk-derived biopeptides have reported in vitro dipeptidyl-peptidase IV (DPP-IV) inhibition, suggesting a glycemic-regulatory effect in Type 2 Diabetes Mellitus (T2DM). Nonetheless, the therapeutic application of these nutraceuticals is limited by the scarcity of knowledge regarding their pharmacokinetic profile. Objective: This study aimed to characterize and assess the pharmacokinetics of milk-derived biopeptides. Through an in silico comparative analysis with gliptins, we expected to identify enhanced properties in food-hydrolysates and suitable DPP-IV inhibiting peptides as candidates for T2DM therapy. Methods: A comparison between gliptins and biopeptides was conducted based on in silico evaluation of drug-likeness, physicochemical properties, pharmacokinetics, and synthetic accessibility. Suitable target proteins for gastrointestinal-absorbable biopeptides were determined as well. Data collection was performed on SwissADME, ADMETlab, DrugBank, SwissTargetPrediction, ChemDes, and BIOPEP-UWM platforms. Statistical analysis was carried out using a one-way ANOVA test. Results: Drug-likeness compliance showed no significant difference between gliptins and biopeptides (p>0.05) in three out of nine assessed rules, though gastrointestinal-absorbable biopeptides exhibited no significant difference with gliptins in five drug-likeness guidelines. The physicochemical evaluation revealed a significant difference (p<0.05) between both groups, with peptides exhibiting enhanced solubility, flexibility, and polarity. Nine out of thirty-six assessed biopeptides reported being likely gastrointestinal-absorbable molecules, from which six displayed ≥30% predicted bioavailability, two reported CYP450 interactions, and all were determined to be blood confined. Biopeptides showed a slightly lower clearance than gliptins yet counteracted by a significantly lower half-life. Moreover, synthetic accessibility scores indicated higher synthetic ease for biopeptides. In addition, absorbable bioactive peptides reported a considerable binding affinity to DPP-IV and Calpain-I. Conclusions: Compared to gliptins, gastrointestinal-absorbable biopeptides exhibit superior physicochemical properties (higher solubility, flexibility, and polarity), lesser CYP450 interactions, higher synthetic ease, and some reported an important affinity for DPP-IV and Calpain-I. Only a small fraction of milk-derived biopeptides are suitable drug-like compounds and feasible candidates for T2DM therapy; yet, testing their therapeutic potency remains subject to further studies


Antecedentes: Los biopéptidos derivados de la leche han mostrado inhibir la dipeptidil-peptidasa IV (DPP-IV) en ensayos in vitro, lo que sugiere una regulación de la glicemia en la Diabetes Mellitus Tipo 2 (DM2). Sin embargo, su uso terapéutico está limitado por el escaso conocimiento de sus propiedades farmacológicas. Objetivo: Caracterizar y evaluar el perfil farmacocinético de los biopéptidos derivados de la leche. Por medio de un análisis comparativo in silico, se buscó identificar propiedades de carácter superior a las gliptinas en los biopéptidos inhibidores de DPP-IV, así como posibles candidatos a agentes terapéuticos en la DMT2. Métodos: Se llevó a cabo una comparación entre las Gliptinas y los biopéptidos basada en la evaluación in silicode las características "d r ug - li ke", propiedades fisicoquímicas, farmacocinética y accesibilidad sintética. Adicionalmente, se determinaron posibles proteínas diana para los biopéptidos de alta probabilidad de absorción gastrointestinal. Los datos se obtuvieron en SwissADME, ADMETlab, DrugBank, SwissTargetPrediction, ChemDes y BIOPEP-UWM. El análisis estadístico se basó en un análisis de varianza (one-way ANOVA test). Resultados: El cumplimiento de las reglas de "drug-likeness" no mostró diferencias significativas entre las gliptinas y los biopéptidos (p>0.05) en tres de las nueve normas evaluadas, empero, los biopéptidos absorbibles no mostraron diferencias significativas con las gliptinas en cinco de estas. La evaluación fisicoquímica reveló una diferencia significativa (p>0.05) entre ambos grupos y una mayor solubilidad, flexibilidad y polaridad para los biopéptidos. Nueve de los treinta y seis biopéptidos estudiados reportaron alta probabilidad de absorción gastrointestinal, de los cuales seis presentaron una biodisponibilidad predicha ≥30%, dos reportaron interacciones con el CYP450, y todos mostraron permanecer confinados en sangre. Los biopéptidos mostraron una tasa de aclaramiento inferior a las gliptinas, sin embargo, contrarrestado por una vida-media significativamente menor. Los valores de accesibilidad sintética indicaron una mayor facilidad de síntesis para los biopéptidos. Por último, los biopéptidos absorbibles mostraron una considerable afinidad por la DPP-IV y la Calpaína-I. Conclusiones: Frente a las gliptinas, los biopéptidos absorbibles presentan: propiedades fisicoquímicas superiores (mayor solubilidad, flexibilidad y polaridad), menores interacciones con el CYP450, mayor facilidad de síntesis y algunos una importante afinidad por la DPP-IV y la Calpaína-I. Una mínima fracción de biopéptidos derivados de la leche son candidatos viables para la terapia de DM2; sin embargo, la determinación de su efectividad terapéutica permanece sujeta a futuros estudios


Subject(s)
Humans , Pharmacokinetics , Peptides , Diabetes Mellitus, Type 2 , Dipeptidyl-Peptidase IV Inhibitors
13.
Electron. j. biotechnol ; 52: 52-58, July. 2021. tab, ilus
Article in English | LILACS | ID: biblio-1283505

ABSTRACT

BACKGROUND: Osteoporosis attacks approximately 10% of the population worldwide. Sika Deer (Cervus nippon), one of China's precious traditional medicinal animals, has been widely recorded in ancient Chinese medical books and claimed for centuries to have numerous medical benefits including bone strengthening. This study aimed to find the use of Sika Deer bone in treating osteoporosis according to traditional records and to investigate the protective effect of Sika Deer bone polypeptide extract on glucocorticoidinduced osteoporosis (GIOP) in rats. RESULTS: Sika Deer bone polypeptide extract could increase serum Ca2+ and BGP, decrease serum P3+, ALP, PTH, and CT, but had no effect on serum NO in rats with GIOP. The immunohistochemical iNOS results of the rats' distal femur were negative in each group. Besides the model group, the eNOS color reaction in osteoblasts was strongly positive in the other three groups. CONCLUSIONS: Sika Deer bone polypeptide extract can improve pathological changes in the microstructure and stimulate the expression of eNOS in osteoblasts. The protective effect on bone might be mediated by eNOS-dependent NO generation.


Subject(s)
Animals , Male , Rats , Osteoporosis/prevention & control , Peptides/pharmacology , Bone and Bones/metabolism , Deer , Osteoblasts , Dexamethasone , Rats, Wistar , Nitric Oxide Synthase Type III/drug effects
14.
Biomédica (Bogotá) ; 41(supl.1): 113-120, mayo 2021. tab
Article in Spanish | LILACS | ID: biblio-1285453

ABSTRACT

Resumen | Introducción. La mayoría de las personas con enfermedad de Chagas desarrolla anticuerpos específicos contra Trypanosoma cruzi. En la infección temprana se producen anticuerpos IgM contra T. cruzi que son reemplazados por IgG durante el curso de la enfermedad. Los primeros síntomas de la enfermedad suelen ser muy leves y atípicos, por lo que a menudo no se detecta en la fase aguda. Objetivos. Evaluar la sensibilidad y la especificidad clínica y analítica, la precisión y la eficacia del UMELISA CHAGAS® con la incorporación de nuevos péptidos sintéticos en la fase sólida representativos de la proteína SAPA (Shed Acute Phase Antigen) y del antígeno TSA (Trypomastigote Surface Antigen). Materiales y métodos. Se evaluó un panel de desempeño de título mixto anti-I cruzi y uno de seroconversión de Chagas, así como muestras de suero positivas y negativas provenientes de zonas endémicas de la enfermedad y muestras positivas de otras enfermedades que podían interferir con la prueba. Las pruebas Bioelisa CHAGAS, Chagatest ELISA recombinante v. 4.0, Chagatest HAI y SD BIOLINE CHAGAS Ab Rapid, se emplearon como referencia. Resultados. Los porcentajes de sensibilidad y especificidad clínica fueron de 97,73 % (IC95% 96,23-99,24) y 99,33 % (IC95% 98,88-99,78), respectivamente. Se obtuvo un 98,96 % de eficacia y una buena precisión. Conclusiones. Los resultados demuestran que la nueva fase sólida del UMELISA CHAGAS® puede utilizarse para el inmunodiagnóstico, la certificación de sangre y la vigilancia epidemiológica en países endémicos y no endémicos con población de alto riesgo.


Abstract | Introduction: Most people with Chagas disease develop specific antibodies against Trypanosoma cruzi. In early infection, IgM antibodies against T. cruzi are produced and later replaced for IgG antibodies during the course of the disease. The first symptoms of the infection may be very mild and atypical, which is why the disease is often not detected in the acute phase. Objectives: To evaluate the clinical and analytical sensitivity, and specificity, accuracy, and efficacy of UMELISA CHAGAS™ with the addition of new synthetic peptides in the solid phase representative of the shed acute phase antigen protein (SAPA) and the trypomastigote surface antigen (TSA). Materials and methods: We evaluated a mixed anti-T. cruzi titer performance panel and a Chagas seroconversion one, as well as positive and negative serum samples from endemic areas of the disease and positive samples for other diseases that may interfere with the assay. The Bioelisa CHAGAS assay, Chaga test recombinant ELISA v.4.0, Chagatest HAI, and SD BIOLINE CHAGAS Ab Rapid were used as reference tests. Results: The sensitivity of the assay was 97.73% (95% CI: 96,23-99,24) and the clinical specificity, 99.33% (95% CI: 98,88-99,78) while the efficacy and the accuracy were 98.96%. Conclusions: Our results show that the new solid phase of UMELISA CHAGAS® can be used for immunodiagnostic, blood certification, and epidemiological surveillance in endemic and non-endemic countries with high-risk populations.


Subject(s)
Chagas Disease/diagnosis , Peptides , Trypanosoma cruzi , Antibodies
15.
Electron. j. biotechnol ; 50: 16-22, Mar. 2021. ilus, tab
Article in English | LILACS | ID: biblio-1292419

ABSTRACT

BACKGROUND: Cecropin P1, acting as an antimicrobial, has a broad-spectrum antibacterial activity with some antiviral and antifungal properties. It is a promising natural alternative to antibiotics which is originally isolated from the pig intestinal parasitic nematode Ascaris suum. Many studies have shown that Cecropin P1 is helpful for the prevention or treatment of clinical diseases. Therefore, it is very necessary to establish a safe, nontoxic, and efficient expression method of Cecropin P1. RESULTS: The results indicated that the recombinant protein was about 5.5 kDa showed by Tricine­SDS­ PAGE and Western blot. And Cecropin P1 was efficiently secreted and expressed after 12 h of induction, with an increasing yield over the course of the induction. Its maximum concentration was 7.83 mg/L after concentration and purification. In addition, in vitro experiments demonstrated that Cecropin P1 not only exerted a strong inhibitory effect on Escherichia coli, Salmonella sp., Shigella sp., and Pasteurella sp., but also displayed an antiviral activity against PRRSV NADC30-Like strain. CONCLUSIONS: Collectively, the strategy of expressing Cecropin P1 in Saccharomyces cerevisiae is harmless, efficient, and safe for cells. In addition, the expressed Cecropin P1 has antiviral and antibacterial properties concurrently.


Subject(s)
Peptides/pharmacology , Saccharomyces cerevisiae/drug effects , Anti-Bacterial Agents/pharmacology , Antiviral Agents/pharmacology , Peptides/chemistry , In Vitro Techniques , Recombinant Proteins , Microbial Sensitivity Tests , Blotting, Western
16.
Article in Chinese | WPRIM | ID: wpr-942303

ABSTRACT

OBJECTIVE@#To assess the activation function of specific tumor polypeptide for dendritic cell vaccine on lymphocytes proliferation, production of cytokines and killing activity in vitro by using dendritic cells as antigen presenting vector.@*METHODS@#Peripheral blood dendritic cells (DC) and cytokine-induced killer (CIK) were isolated and cultured by adherent culture method; CCK-8 method was used to assess the proliferation function of lymphocytes and the killing function of lymphocytes to tumor cells; enzyme-linked immunospot assay method was used to evaluate the secretion function of cytokines. The experiment was divided into tumor polypeptide group (peptide with DC-CIK), DC-CIK group and CIK group.@*RESULTS@#With presence of interleukin-2 (IL-2) in the culture system, the lymphocyte proliferation of the three groups was obvious. The absorbance at 450 nm of tumor polypeptide group was significantly higher than that of CIK group at the time points day 4 and day 6 (day 4: Z=-3.79, P < 0.001; day 6: Z =-2.95, P < 0.01). The absorbance at 450 nm of group tumor polypeptide was significantly higher than that of DC-CIK group on day 4 (Z=-2.02, P < 0.05). Without IL-2 in the culture system, lymphocytes proliferated slowly in all the three groups, and there was no significant difference in 450 nm absorbance at each time point. The levels of IL-4 (Z=-2.61, P < 0.01), granulocyte-macrophage colony-stimulation factor (GM-CSF, Z=-3.85, P < 0.001), interferon- γ (IFN- γ, Z=-3.56, P < 0.001) and tumor necrosis factor-α (TNF-ɑ, Z=-3.40, P < 0.001) of tumor polypeptide group were higher than those of CIK group. There was no significant difference in the production of cytokines except IL-4 (Z=-2.15, P < 0.05) when tumor polypeptide group was compared with DC-CIK group. The production of IFN-γ (Z=-2.44, P < 0.05), TNF-ɑ (Z=-2.26, P < 0.05) and GM-CSF (Z=-3.73, P < 0.001) in DC-CIK group were higher than those of CIK group. Although there was no significant difference in killing activity between tumor polypeptide group, DC-CIK group and CIK group at hour 18 and hour 24, and the killing activity of tumor polypeptide group was higher than that of the other two groups.@*CONCLUSION@#Tumor peptide combined with dendritic cells can improve the proliferation activity of CIK cells in vitro, and increase the secretion of several cytokines.


Subject(s)
Dendritic Cells , Peptides
17.
Braz. j. med. biol. res ; 54(5): e10717, 2021. tab, graf
Article in English | LILACS | ID: biblio-1180740

ABSTRACT

Scorpion venom is a Chinese medicine for epilepsy treatment, but the underlying mechanism is not clear. Scorpion venom heat-resistant peptide (SVHRP), a peptide isolated from the venom of Buthus martensii Karsch, has an anti-epileptic effect by reducing seizure behavior according to a modified Racine scale. The present study aimed to investigate the molecular mechanism of SVHRP on temporal lobe epilepsy. The hippocampus and hippocampal neurons from kainic acid-induced epileptic rats were treated with SVHRP at different doses and duration. Quantitative RT-PCR and immunoblotting were used to detect the expression level of brain-derived neurotrophic factor (BDNF), neuropeptide Y (NPY), cAMP-response element binding protein (CREB), stromal interaction molecule (STIM), and calcium release-activated calcium channel protein 1 (ORAI1). In the hippocampal tissues and primary hippocampal neuron cultures, SVHRP treatment resulted in increased mRNA and protein levels of BDNF and NPY under the epileptic condition. The upregulation of BDNF and NPY expression was positively correlated with the dose level and treatment duration of SVHRP in hippocampal tissues from kainic acid-induced epileptic rats. On the other hand, no significant changes in the levels of CREB, STIM, or ORAI1 were observed. SVHRP may exhibit an anti-epileptic effect by upregulating the expression of BDNF and NPY in the epileptic hippocampus.


Subject(s)
Animals , Rats , Scorpion Venoms/toxicity , Epilepsy/chemically induced , Epilepsy/drug therapy , Peptides , Brain-Derived Neurotrophic Factor/metabolism , Hot Temperature , Hippocampus/metabolism , Kainic Acid/toxicity , Neurons
18.
Braz. J. Pharm. Sci. (Online) ; 57: e19061, 2021. tab, graf
Article in English | LILACS | ID: biblio-1350245

ABSTRACT

Proteins and peptides are the most diverse biomolecules found in nature and make our interest due to their wide applications in food and pharmaceutical industry. Angiotensin Converting Enzyme (ACE) plays a major role in controlling blood pressure. The inhibition of ACE with peptides is a main target in the regulation of hypertension. The objective of the present study was to investigate the therapeutic potential of soy bean. This was accomplished by isolation of ACE inhibitory peptides using response surface methodology (RSM) and characterization of these bioactive peptides by mass spectrometry. 31 hydrolyzed fractions were isolated and evaluated for their ACE inhibition potential. Hydrolyzed fraction having highest ACE inhibitory activity was characterized by liquid chromatography-mass spectrometry (LC-MS) technique. RSM results showed maximum ACE inhibition potential (64%) by hydrolyzate was obtained at 45 ºC temperature, pH 8.0, E/S 0.2 in 2 hours hydrolysis time. Results of LC-MS analysis revealed Ser-Gly, Ser-Pro, Met-Ala, His-Ala, Lys-Pro, Phe-Thr, Met-Leu, Pro-Arg, Ala-Pro-Val, Pro-Ala-Leu, Val-Met-Gly, Pro-Leu-Val, Pro-Pro-Gln, His-Arg-Gly, Ser-Phe-Val-Leu, Ala-Val-His-Try, Arg-Thr-Val-Arg, His-His-Tyr-Leu-Val, Asp-Gly-Ala-Cys-Ser-Ala-Asn and MetVal-Thr-Gly-Pro-Gly-Cys-His bioactive peptides in hydrolyzed fraction of soy bean. Our data provide evidence that response surface methodology is a good approach for isolation of antihypertensive bioactive peptides with more potent activity as nutraceuticals or pharmaceuticals. Therefore soy bean can be use for industrial production of pharmaceutical grade natural medicines for handling high blood pressure.


Subject(s)
Peptides/pharmacology , Proteins/pharmacology , Soybean Proteins/pharmacology , Dietary Supplements , Protein Hydrolysates/pharmacology , Mass Spectrometry , Chromatography, Liquid/methods , Process Optimization/classification , Hydrogen-Ion Concentration , Hypertension/therapy , Antihypertensive Agents/analysis
19.
Chinese Journal of Biotechnology ; (12): 2915-2923, 2021.
Article in Chinese | WPRIM | ID: wpr-887853

ABSTRACT

Antimicrobial peptides are the most promising alternatives to antibiotics. However, the strategy of producing antimicrobial peptides by recombinant technology is complicated and expensive, which is not conducive to the large-scale production. Oxysterlin 1 is a novel type of cecropin antimicrobial peptide mainly targeting on Gram-negative bacteria and is of low cytotoxicity. In this study, a simple and cost-effective method was developed to produce Oxysterlin 1 in Escherichia coli. The Oxysterlin 1 gene was cloned into a plasmid containing elastin-like polypeptide (ELP) and protein splicing elements (intein) to construct the recombinant expression plasmid (pET-ELP-I-Oxysterlin 1). The recombinant protein was mainly expressed in soluble form in E. coli, and then the target peptide can be purified with a simple salting out method followed by pH changing. The final yield of Oxysterlin 1 was about 1.2 mg/L, and the subsequent antimicrobial experiment showed the expected antimicrobial activity. This study holds promise for large-scale production of antimicrobial peptides and the in-depth study of its antimicrobial mechanism.


Subject(s)
Elastin , Escherichia coli/genetics , Inteins , Peptides/pharmacology , Pore Forming Cytotoxic Proteins , Recombinant Fusion Proteins/genetics
20.
Chinese Journal of Biotechnology ; (12): 2334-2341, 2021.
Article in Chinese | WPRIM | ID: wpr-887800

ABSTRACT

Tyrosine phosphorylation is one of the important protein phosphorylations in eukaryotes responsible for a variety of biological processes including cell signaling transduction, cell migration, and apoptosis. In the study of phosphoproteomics, due to the low stoichiometry of tyrosine phosphorylation (pTyr) proteins and sometimes limited initial sample, traditional phosphoproteomics enrichment technology is inefficient for the enrichment of pTyr peptides. Here, we review the substantial progress in tyrosine phosphoproteomics by preparation of limited amount sample and the newly introduced SH2 superbinder.


Subject(s)
Cell Movement , Peptides , Phosphorylation , Technology , Tyrosine/metabolism
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