RESUMO
Starch is a promising polymer for creating novel microparticulate systems with superior biocompatibility and controlled drug delivery capabilities. In this study, we synthesized polyethylene glycol (PEG)-modified starch microparticles and encapsulated folic acid using a solvent-mediated acid-base precipitation method with magnetic stirring, which is a simple and effective method. To evaluate particle degradation, we simulated physiological conditions by employing an enzymatic degradation approach. Our results with FTIR and SEM confirmed the successful synthesis of starch-PEG microparticles encapsulating folic acid. The average size of starch microparticles encapsulating folic acid was 4.97 µm and increased to 6.01 µm upon modification with PEG. The microparticles were first exposed to amylase at pH 6.7 and pepsin at pH 1.5 at different incubation times at physiological temperature with shaking. Post-degradation analysis revealed changes in particle size and morphology, indicating effective enzymatic degradation. FTIR spectroscopy was used to assess the chemical composition before and after degradation. The initial FTIR spectra displayed characteristic peaks of starch, PEG, and folic acid, which showed decreased intensities after enzymatic degradation, suggesting alterations in chemical composition. These findings demonstrate the ongoing development of starch-PEG microparticles for controlled drug delivery and other biomedical applications and provide the basis for further exploration of PEG-starch as a versatile biomaterial for encapsulating bioactive compounds.
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In this article, the synthesis of antioxidant peptides in the enzymatic hydrolysis of caprine casein was analyzed at three different time points (60 min, 90 min, and 120 min) using immobilized pepsin on activated and modified carbon (AC, ACF, ACG 50, ACG 100). The immobilization assays revealed a reduction in the biocatalysts' activity compared to the free enzyme. Among the modified ones, ACG 50 exhibited greater activity and better efficiency for reuse cycles, with superior values after 60 min and 90 min. Peptide synthesis was observed under all studied conditions. Analyses (DPPH, ß-carotene/linoleic acid, FRAP) confirmed the antioxidant potential of the peptides generated by the immobilized enzyme. However, the immobilized enzyme in ACG 50 and ACG 100, combined with longer hydrolysis times, allowed the formation of peptides with an antioxidant capacity greater than or equivalent to those generated by the free enzyme, despite reduced enzymatic activity.
Assuntos
Antioxidantes , Caseínas , Enzimas Imobilizadas , Glutaral , Cabras , Iridoides , Pepsina A , Peptídeos , Antioxidantes/química , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Caseínas/química , Animais , Pepsina A/metabolismo , Pepsina A/química , Glutaral/química , Peptídeos/química , Iridoides/química , Hidrólise , Carvão Vegetal/químicaRESUMO
Increased prevalence of diabetes prompts the development of foods with reduced starch digestibility. This study analyzed the impact of adding soluble dietary fiber (inulin-IN; polydextrose-PD) to baked gluten-starch matrices (7.5-13%) on microstructure formation and in vitro starch digestibility. IN and PD enhanced water-holding capacity, the hardness of baked matrices, and lowered water activity in the formulated matrices, potentially explaining the reduced starch gelatinization degree as IN or PD concentration increased. A maximum gelatinization decrease (26%) occurred in formulations with 13% IN. Micro-CT analysis showed a reduction in total and open porosity, which, along with the lower gelatinization degree, may account for the reduced in vitro starch digestibility. Samples with 13% IN exhibited a significantly lower rapidly available glucose fraction (8.56 g/100 g) and higher unavailable glucose fraction (87.76 g/100 g) compared to the control (34.85 g/100 g and 47.59 g/100 g, respectively). These findings suggest the potential for developing healthier, starch-rich baked foods with a reduced glycemic impact.
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The INFOGEST protocol has been widely used as a static in-vitro simulation of gastrointestinal food digestion for bioaccessibility assessments on bioactive compounds. The standardization of the activity of several enzymes, such as pepsin, via UV-spectrophotometry of digested hemoglobin at 280 nm is a key step in the protocol. Standardization is a crucial stage since it is necessary to determine the quantity of enzyme to be added to the sample for digestion. However, this method is yet to be analytically validated; it requires quartz cuvettes and large volumes of samples and is time-consuming. Thus, we reviewed and adapted a well-known colorimetric method in microplates array by using the Folin-Ciocalteu reagent, and this study is the first to report for miniaturization of this method, the advantages of which include its automation, ease of use, the low volume of samples required, the minimal use of reagents, and speed. This method was compared to the traditional UV method, and the comparison results show no statistical difference between the inter day means for each group (p > 0.05). The proposed method was validated, showing high reproducibility (8% as inter-day CV) and statistically comparable results with the traditional UV spectrophotometric method.
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Medicinal plants offer a valuable source of natural compounds with specific and selective bioactivity. These compounds have been isolated since the mid-nineteenth century and are now commonly used in modern medications. L. octovalvis (Jacq.) P.H.Raven, C. aconitifolius, and C. longirostrata are Mexican medicinal plants consumed regularly, and research has shown that they contain bioactive compounds capable of promoting the inhibition of digestive enzymes. This is noteworthy since enzyme inhibitors are bioactive substances that interact with enzymes, diminishing their activity and thereby contributing to the management of diseases and metabolic disturbances. To investigate the activity of these plants, individual analyses were conducted, assessing their proximal composition, bioactive compounds, and inhibition of α-Amylase, α-Glucosidase, lipase, and pepsin. The results revealed that all three plants exhibited enzymatic inhibition. When comparing the plants, it was determined that C. aconitifolius had the lowest concentration required for a 50% inhibition in α-Amylase, α-Glucosidase, and lipase, as indicated by the IC50 values. For pepsin, C. longirostrata demonstrated the lowest IC50 value. By understanding the bioactive compounds present in these plants, we can establish the relationship they have with enzymatic inhibition, which can be utilized for future investigations.
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Abstract Objective: Salivary pepsin has emerged as a biomarker for Laryngopharyngeal Reflux (LPR), which, however, has been questioned for its efficacy due to a lack of supporting medical data. Therefore, this study analyzed the diagnostic value of salivary pepsin for LPR and assessed a better cutoff value. Methods: Studies were searched in PubMed, Embase, and Cochrane Library from their receptions to October 1, 2021. Then, RevMan 5.3 and Stata 14.0 were utilized to summarize the diagnostic indexes for further meta-analysis. Data were separately extracted by two reviewers according to the trial data extraction form of the Cochrane Handbook. The risk of bias in Randomized Control Trials (RCTs) was evaluated with the Cochrane Risk of Bias Tool. Results: A total of 16 studies matched the criteria and were subjected to meta-analysis. The results revealed a pooled sensitivity of 61% (95% CI 50%-71%), a pooled specificity of 67% (95% CI 48%-81%), a positive likelihood ratio of 2 (95% CI 1.2-2.8), a negative likelihood ratio of 0.58 (95% CI 0.47-0.72), and the area under the receiver operating characteristic curve of 0.67 (95% CI 0.63-0.71). Subgroup analyses indicated that the cutoff value of pepsin at 50 ng/mL had a higher degree of diagnostic accuracy than that of pepsin at 16 ng/mL in cohort studies. Conclusion: The review demonstrated low diagnostic performance of salivary pepsin for LPR and that the cutoff value of 50 ng/mL pepsin had superior diagnostic accuracy. Nevertheless, the diagnostic value may vary dependent on the utilized diagnostic criteria. Therefore, additional research is needed on the improved way of identifying salivary pepsin in the diagnosis of LPR, and also longer-term and more rigorous RCTs are warranted to further assess the effectiveness of salivary pepsin.
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SUMMARY: The aim of the present in vitro study is to visualize dentin to get an in-depth knowledge of the nature of dentin that could provide useful information regarding conditioning dentinal substrate when treating dentinal lesions. Forty-nine extracted human third molars were obtained and prepared to produce artificial dentinal lesions through demineralizing with acetic acid for 7 and 14 days, or lactic acid for 7 days. The teeth were divided into groups and treated with either NaOCl, pepsin, trypsin, or phosphoric acid. To obtain information on the morphology of the treated dentinal surfaces, all samples were visualized under high resolution field emission scanning electron microscope. With high magnification reaching x50000 dentin was clearly visualized together with its constitutes. The effect of various demineralization approaches and various treatment protocols were demonstrated clearly. The relationship between the conditioning procedure steps and the subsequent bond strength was discussed. To our best knowledge, there is no previous clear highly magnified scanning electron microscope images for dentin, and dentinal components and constitutes with and without various treatments. The current in vitro study suggests the complexity nature of dentin as a substrate that should be treated carefully especially with technique sensitive procedures such as adhesive restorations.
El objetivo del presente estudio in vitro fue visualizar la dentina para obtener un conocimiento completo de la naturaleza de ella lo que podría proporcionar información útil sobre el acondicionamiento del sustrato dentinario en el tratamiento de lesiones dentinarias. Se obtuvieron 49 terceros molares humanos extraídos y se prepararon para producir lesiones dentinales artificiales mediante desmineralización con ácido acético por 7 y 14 días, o ácido láctico por 7 días. Los dientes se dividieron en grupos y se trataron con NaOCl, pepsina, tripsina o ácido fosfórico. Para obtener información sobre la morfología de las superficies dentinarias tratadas, todas las muestras se visualizaron bajo un microscopio electrónico de barrido de emisión de campo de alta resolución. Con un gran aumento que alcanzó x50000, la dentina se visualizó claramente junto con sus componentes. Se demostró el efecto de varios enfoques de desmineralización y varios protocolos de tratamiento. Se discutió la relación entre los pasos del procedimiento de acondicionamiento y la subsiguiente fuerza de unión. Hasta donde sabemos, no hay imágenes claras previas de microscopio electrónico de barrido altamente ampliadas para la dentina y los componentes y constituyentes de la dentina con y sin diferentes tratamientos. El estudio in vitro actual sugiere la naturaleza compleja de la dentina como sustrato que debe tratarse con cuidado, especialmente en los procedimientos sensibles a la técnica, tal como las restauraciones adhesivas.
Assuntos
Humanos , Desmineralização do Dente/induzido quimicamente , Dentina/efeitos dos fármacos , Dentina/ultraestrutura , Hipoclorito de Sódio , Microscopia Eletrônica de Varredura , Tripsina , Pepsina A , Ácido Acético/farmacologia , Ácido Láctico/farmacologiaRESUMO
OBJECTIVES: Salivary pepsin has emerged as a biomarker for Laryngopharyngeal Reflux (LPR), which, however, has been questioned for its efficacy due to a lack of supporting medical data. Therefore, this study analyzed the diagnostic value of salivary pepsin for LPR and assessed a better cutoff value. METHODS: Studies were searched in PubMed, Embase, and Cochrane Library from their receptions to October 1, 2021. Then, RevMan 5.3 and Stata 14.0 were utilized to summarize the diagnostic indexes for further meta-analysis. Data were separately extracted by two reviewers according to the trial data extraction form of the Cochrane Handbook. The risk of bias in Randomized Control Trials (RCTs) was evaluated with the Cochrane Risk of Bias Tool. RESULTS: A total of 16 studies matched the criteria and were subjected to meta-analysis. The results revealed a pooled sensitivity of 61% (95% CI 50%-71%), a pooled specificity of 67% (95% CI 48%-81%), a positive likelihood ratio of 2 (95% CI 1.2-2.8), a negative likelihood ratio of 0.58 (95% CI 0.47â0.72), and the area under the receiver operating characteristic curve of 0.67 (95% CI 0.63â0.71). Subgroup analyses indicated that the cutoff value of pepsin at 50â¯ng/mL had a higher degree of diagnostic accuracy than that of pepsin at 16â¯ng/mL in cohort studies. CONCLUSION: The review demonstrated low diagnostic performance of salivary pepsin for LPR and that the cutoff value of 50â¯ng/mL pepsin had superior diagnostic accuracy. Nevertheless, the diagnostic value may vary dependent on the utilized diagnostic criteria. Therefore, additional research is needed on the improved way of identifying salivary pepsin in the diagnosis of LPR, and also longer-term and more rigorous RCTs are warranted to further assess the effectiveness of salivary pepsin.
Assuntos
Refluxo Laringofaríngeo , Humanos , Refluxo Laringofaríngeo/diagnóstico , Pepsina A/análise , Saliva , Curva ROC , BiomarcadoresRESUMO
The viscera and other residues from fish processing are commonly discarded by the fishing industry. These byproducts can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.
As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-1. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.
Assuntos
Peptídeo Hidrolases , Estômago , Temperatura , Concentração de Íons de HidrogênioRESUMO
Abstract The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by-products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 Umg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.
Resumo As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 Umg-1. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas
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Objective: To evaluate the efficacy of a new pepsin enzyme-based gel compared with Carisolv as a CMCR agent. Clinical and radiographical evaluations of recurrent caries were made 3 and 6 months after treatment. Material and Methods: A split-mouth designed randomized controlled clinical study was carried out on 40 primary anterior teeth of children aged between 4-7 years. Pepsin enzyme-based gel and Carisolv solution were applied to carious lesions until complete removal of caries. The efficacy of both agents was evaluated by the number of application times to remove all caries. Recurrent caries were evaluated clinically and radiographically after 3 and 6 months of treatment. Results: Results showed no statistically significant differences in the efficacy of caries removal by the number of application times (P = 0.919). Concerning recurrent caries, clinical and radiographical evaluation after three and six months showed no statistically significant differences between the two groups (P = 0.574, P = 0.547, respectively). Conclusion: Pepsin enzyme-based gel can be considered similar to Carisolv gel regarding its efficacy as a CMCR agent for small carious lesions on primary anterior teeth in children aged 4-7 years old. (AU)
Objetivo: avaliar a eficácia de um novo gel a base de enzima pepsina comparada com o Carisolv como um agente na remoção químico-mecânica da cárie. Avaliações clínicas e radiográficas de cárie recorrente foram feitas em 3 e 6 meses apos o tratamento. Material e Métodos: um estudo clínico controlado randomizado de boca-dividida foi realizado em 40 dentes deciduos anteriores de crianças com idade entre 4-7 anos. Gel à base de enzima pepsina e a soluçao de Carisolv foram aplicados sobre a lesão cariosa até a completa remoção da carie. A eficácia de ambos agentes foi avaliada pelo número de tempo de aplicações para a remoção de todo tecido cariado. Cárie recorrente foi avaliada clinicamente e radiograficamente após 3 e 6 meses de tratamento. Resultados: Não houve diferença significativa na eficácia de remoção de cárie pelo número de tempo de aplicação (P = 0.919). Em relação à cárie recorrente, avaliação clínica e radiográfica apos 3 e 6 meses mostraram que não houve diferença estatisticamente significante entre os 2 grupos (P = 0.574, P = 0.547, respectivamente). Conclusão: o gel à base de enzima pepsina pode ser considerado similar ao gel Carisolv em relação a sua eficácia como um agente químico-mecânico na remoção da cárie para lesões cariosas pequenas em dentes anteriores decíduos em crianças entre 4-7 anos de idade.(AU)
Assuntos
Humanos , Criança , Pepsina A , Cárie Dentária , OdontologiaRESUMO
The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-¹. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.
As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-¹. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.
Assuntos
Animais , Colágeno/análise , Estômago , Pepsina A/análise , Perciformes , Vísceras/enzimologia , Ácido Aspártico Proteases/análiseRESUMO
The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-¹. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.(AU)
As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-¹. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.(AU)
Assuntos
Animais , Ácido Aspártico Proteases/análise , Vísceras/enzimologia , Estômago , Pepsina A/análise , Colágeno/análise , PerciformesRESUMO
Jackfruit (Artocarpus heterophyllus Lam.) is an evergreen tree that produces a high waste of leaves. This study evaluated the obtention of peptides from jackfruit leaves using pancreatin and pepsin, their antifungal activity, and their effect on pectin films. The protein content was 7.64 ± 0.12 g/100 g of jackfruit fresh leaves. Pancreatin produced a higher yield than pepsin in the obtention of peptides (p ≤ 0.05). However, peptides obtained after 2 h by pepsin hydrolysis (Pep-P) had six essential amino acids and inhibited > 99% of mycelial growth and spore germination of Colletotrichum gloeosporioides. Pectin films with Pep-P showed a slight brown color, lower thickness, water vapor permeability, and moisture content, as well as higher thermal stability and better inhibition properties against C. gloeosporioides than pectin films without Pep-P (p ≤ 0.05). Pectin films added with Pep-P from jackfruit leaf could be a green alternative to anthracnose control in tropical fruits.
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Introduction Twenty-four-hour multichannel intraluminal impedance with double probe pH monitoring (MII-pH), though considered the most sensitive tool for the diagnosis of gastroesophageal reflux disease (GERD), is invasive, time consuming, not widely available, and unable to detect non-acid reflux. In contrast, the presence of pepsin in the saliva would act as a marker for reflux, considering that pepsin is only produced in the stomach. Objective To evaluate the predictive value of salivary pepsin in diagnosing laryngopharyngeal reflux (LPR) as suggested by the results of reflux symptom index (RSI > 13), reflux finding score (RFS > 7), and positive response to treatment with a 4-week course of proton-pump inhibitors. Methods This prospective study was done at a tertiary care hospital on 120 adult patients attending ENT OPD with clinical diagnosis of LPR. The presence of pepsin in their pharyngeal secretions and saliva using a lateral flow device, the Peptest, was compared with RSI, RFS, and with the response to medical treatment using the Chi-squared test. Results Salivary pepsin was found to be positive in 68% of the patients, with 87.5% of them showing positive response to treatment. Chi-squared analysis showed a significant association between positive salivary pepsin and RFS > 7, RSI >13, a combination of RFS > 7 and RSI > 13 as well as with response to treatment ( p < 0.0001). Conclusion When considered along with the clinical indicators of RFS and RSI of more than 7 and 13, respectively, and/or with a response to treatment, a positive salivary pepsin test indicates statistically significant chance of presence of LPR.
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Abstract Introduction Twenty-four-hour multichannel intraluminal impedance with double probe pH monitoring (MII-pH), though considered the most sensitive tool for the diagnosis of gastroesophageal reflux disease (GERD), is invasive, time consuming, not widely available, and unable to detect non-acid reflux. In contrast, the presence of pepsin in the saliva would act as a marker for reflux, considering that pepsin is only produced in the stomach. Objective To evaluate the predictive value of salivary pepsin in diagnosing laryngopharyngeal reflux (LPR) as suggested by the results of reflux symptom index (RSI > 13), reflux finding score (RFS > 7), and positive response to treatment with a 4-week course of proton-pump inhibitors. Methods This prospective study was done at a tertiary care hospital on 120 adult patients attending ENT OPD with clinical diagnosis of LPR. The presence of pepsin in their pharyngeal secretions and saliva using a lateral flow device, the Peptest, was compared with RSI, RFS, and with the response to medical treatment using the Chi-squared test. Results Salivary pepsin was found to be positive in 68% of the patients, with 87.5% of them showing positive response to treatment. Chi-squared analysis showed a significant association between positive salivary pepsin and RFS > 7, RSI >13, a combination of RFS > 7 and RSI > 13 as well as with response to treatment (p < 0.0001). Conclusion When considered along with the clinical indicators of RFS and RSI of more than 7 and 13, respectively, and/or with a response to treatment, a positive salivary pepsin test indicates statistically significant chance of presence of LPR.
RESUMO
ABSTRACT: The effect of high hydrostatic pressure (HHP) application on whey protein concentrate was evaluated both before (pre-treatment - PT) and during (hydrolysis assisted - HA) hydrolysis processes. A factorial design 22 with 3 central points was used with pressure (100, 250, 400 MPa) and time (5, 20 and 35 minutes) as independent variables. The hydrolysis was evaluated and monitored by soluble protein, aromatic amino acid contents and RP-HPLC. ABTS and ORAC tests were used to evaluate the in vitro antioxidant capacity. The reduction of soluble protein content was approximately 20% for conventional hydrolysis and for all PT treatments up to 4 h of reaction, while HHP assisted hydrolysis at 100 MPa showed a 35% protein reduction after 35 minutes of reaction. In addition, pressurization favored peptic hydrolysis of β-lactoglobulin by up to 98% and also improved the in vitro antioxidant capacity of the hydrolysates, which increased from 34.25 to 60.89 μmoles TE g-1 of protein in the best treatment. The results suggest that the use of HHP assisted hydrolysis favored the peptic hydrolysis, with a reduction in hydrolysis time and increased antioxidant activity.
RESUMO: Neste estudo, o efeito da aplicação de alta pressão hidrostática (HHP) sobre o concentrado proteico de soro de leite foi avaliado antes (pré-tratamento - PT) e durante os processos de hidrólise (assistida por hidrólise - HA). Utilizou-se o delineamento fatorial 22 com três pontos centrais, onde as variáveis independentes foram pressão (100, 250, 400 MPa) e tempo (5, 20 e 35 minutos). A hidrólise foi avaliada pelo conteúdo de proteínas solúveis e aminoácidos aromáticos, além do perfil peptídico por RP-HPLC. As análises de ABTS e ORAC foram utilizadas para avaliar a capacidade antioxidante in vitro. A redução do teor de proteína solúvel foi de aproximadamente 20% para a hidrólise convencional e para todos os pontos de PT até 4h de reação, enquanto a hidrólise assistida por HHP a 100 MPa mostrou uma redução de 35% de proteína em 35 minutos de reação. Além disso, a pressurização favoreceu a hidrólise péptica da β-lactoglobulina em até 98% e também melhorou a capacidade antioxidante in vitro dos hidrolisados, que aumentaram de 34,25 para 60,89 μmoles de TE g-1 de proteína no melhor tratamento. Os resultados sugerem que o uso da hidrólise assistida por HHP favoreceu a hidrólise péptica, com redução no tempo de hidrólise e aumento da atividade antioxidante.
RESUMO
The effect of high hydrostatic pressure (HHP) application on whey protein concentrate was evaluated both before (pre-treatment - PT) and during (hydrolysis assisted - HA) hydrolysis processes. A factorial design 22 with 3 central points was used with pressure (100, 250, 400 MPa) and time (5, 20 and 35 minutes) as independent variables. The hydrolysis was evaluated and monitored by soluble protein, aromatic amino acid contents and RP-HPLC. ABTS and ORAC tests were used to evaluate the in vitro antioxidant capacity. The reduction of soluble protein content was approximately 20% for conventional hydrolysis and for all PT treatments up to 4 h of reaction, while HHP assisted hydrolysis at 100 MPa showed a 35% protein reduction after 35 minutes of reaction. In addition, pressurization favored peptic hydrolysis of -lactoglobulin by up to 98% and also improved the in vitro antioxidant capacity of the hydrolysates, which increased from 34.25 to 60.89 moles TE g-1 of protein in the best treatment. The results suggest that the use of HHP assisted hydrolysis favored the peptic hydrolysis, with a reduction in hydrolysis time and increased antioxidant activity.(AU)
Neste estudo, o efeito da aplicação de alta pressão hidrostática (HHP) sobre o concentrado proteico de soro de leite foi avaliado antes (pré-tratamento - PT) e durante os processos de hidrólise (assistida por hidrólise - HA). Utilizou-se o delineamento fatorial 22 com três pontos centrais, onde as variáveis independentes foram pressão (100, 250, 400 MPa) e tempo (5, 20 e 35 minutos). A hidrólise foi avaliada pelo conteúdo de proteínas solúveis e aminoácidos aromáticos, além do perfil peptídico por RP-HPLC. As análises de ABTS e ORAC foram utilizadas para avaliar a capacidade antioxidante in vitro. A redução do teor de proteína solúvel foi de aproximadamente 20% para a hidrólise convencional e para todos os pontos de PT até 4h de reação, enquanto a hidrólise assistida por HHP a 100 MPa mostrou uma redução de 35% de proteína em 35 minutos de reação. Além disso, a pressurização favoreceu a hidrólise péptica da -lactoglobulina em até 98% e também melhorou a capacidade antioxidante in vitro dos hidrolisados, que aumentaram de 34,25 para 60,89 moles de TE g-1 de proteína no melhor tratamento. Os resultados sugerem que o uso da hidrólise assistida por HHP favoreceu a hidrólise péptica, com redução no tempo de hidrólise e aumento da atividade antioxidante.(AU)
Assuntos
Proteínas do Soro do Leite/antagonistas & inibidores , Proteínas do Soro do Leite/análise , Enzimas/farmacocinética , Especificidade por SubstratoRESUMO
Synthesis of nanocomplexes is a simple and low-cost technique for the production of encapsulation systems aiming industrial applications, based on the interaction of at least two oppositely charged molecules. Gellan gum (anionic) is a water-soluble biopolymer resistant to stomach pH conditions, therefore an interesting alternative as an encapsulating matrix. Chitosan (cationic) is also widely used due to its biocompatibility and mucoadhesive properties, although its low water solubility is an important step to be overcome for the production of the complexes. To improve this property, many techniques have been employed, but most of them use unsustainable techniques and chemical agents. The enzymatic hydrolysis of chitosan using proteases emerges as an alternative to these drawbacks and, therefore, this study aimed to evaluate the electrostatic nanocomplexation of native (C) or hydrolyzed (HC) chitosan (by porcine pepsin protease) with gellan gum (G). Polysaccharides and nanocomplexes formed with different G:C or G:HC ratio were evaluated by zeta potential measurements, particle size distribution, X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Transmission Electron Microscopy (STEM), intrinsic viscosity and turbidity analyses. Chitosan hydrolysis allowed the formation of a smaller (445.3 nm in pH 4.5) and more soluble structure (3 kDa), which positively influenced the formation of the complexes. The ratios G:HC of 7:3 and 8:2 formed complexes with lower values of zeta potential (13.9 mV and -5.0 mV, respectively), particle size (635.8 nm and 533.6 nm, respectively) and polydispersity (0.28 and 0.23) compared to complexes formed with native chitosan. Overall, our results show that enzymatic hydrolysis of chitosan favored the formation of electrostatic complexes with reduced size and low polydispersity, which can be used as efficient encapsulating matrices for improved targeted delivery and controlled release of bioactive compounds.
Assuntos
Quitosana , Nanopartículas , Animais , Hidrólise , Tamanho da Partícula , Solubilidade , SuínosRESUMO
Most antivenoms are produced by techniques developed over 50 years ago, with minor modifications. Herein we revise the core of traditional antivenom production processes aiming to optimize key determinants for both consistent antivenom production and the best balance between F(ab')2 quality and recovery. Factorial design analysis revealed that pepsin digestion of 1:3 saline diluted equine plasma for 60 min under pH: 3.20, 37 °C temperature and a 1:15 pepsin to protein ratio conditions, allowed to achieve maximal IgG to F(ab')2 conversion with minimal protein aggregate formation. Further downstream processing by salting out with ammonium sulfate was also studied by factorial analysis. The influence of ammonium sulfate (AS) concentration, temperature (T) and the albumin to total plasma protein ratio plasma (Alb:P) were assayed, revealing that both AS, T and their interaction have a significant impact in F(ab')2 quality and recovery. Taking into account the existing compromise between F(ab')2 monomer recovery and quality two alternative conditions were selected: 14 g/dl AS at 56 °C and, alternatively 16 g/dl AS at 30 °C. Reasonable yields (42%) and product quality (2.5% of aggregates) without significant changes in production cost of traditional methodologies was achieved under the optimized conditions found.