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The objective of this work was to optimize the application of an enzymatic blend produced by Aspergillus niger ATCC 1004 on the Pimenta dioica fruits for essential oil extraction. The enzyme blend was obtained from the fermentation of cocoa bean shells, an agro-industrial residue. The effects of the enzymatic pre-treatment on the extraction yield, the chemical composition of the oil through gas chromatography, and the fruit structure through scanning electron microscopy (SEM) were assessed. A Doehlert design was used to optimize the process conditions, resulting in an extraction with 117 mL of enzyme during 77 min, which increased the extraction yield by 387.5%. The chemical composition was not altered, which proves that the enzyme blend preserves the quality of the essential oil extracted. The content of eugenol (70%), the major compound in the P. dioica essential oil, had a great increase in its concentration (560%). The enzyme activity analyses showed the presence of endoglucanase (0.4 U/mL), exoglucanase (0.25 U/mL), ß-glucosidase (0.19 U/mL), and invertase (135.08 U/mL). The microscopy analyses revealed changes in the morphology of fruit surface due to the enzymatic action. These results demonstrate the great potential of using enzyme blends produced by filamentous fungi from agro-industrial residues for the essential oils extraction of interest for the pharmaceutical and food industries.

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This study presents a novel approach to producing activated carbon from agro-industrial residues, specifically cocoa fruit peel, using solid-state fermentation (SSF) with Aspergillus niger. The process effectively degrades lignin, a major impediment in traditional activated carbon production, resulting in a high-quality carbon material. This carbon was successfully utilized for enzyme immobilization and aroma synthesis, showcasing its potential as a versatile biocatalyst. The study meticulously evaluated the physical and chemical attributes of activated carbon derived from fermented cocoa peel, alongside the immobilized enzymes. Employing a suite of analytical techniques-electrophoresis, FTIR, XRD, and TG/DTG the research revealed that fermentation yields a porous material with an expansive surface area of 1107.87 m2/g. This material proves to be an excellent medium for lipase immobilization. The biocatalyst fashioned from the fermented biomass exhibited a notable increase in protein content (13% w/w), hydrolytic activity (15% w/w), and specific activity (29% w/w), underscoring the efficacy of the fermentation process. The significant outcome of this research is the development of a sustainable method for activated carbon production that not only overcomes the limitations posed by lignin but also enhances enzyme immobilization for industrial applications. The study's findings have important implications for the agro-industrial sector, promoting a circular economy and advancing sustainable biotechnological processes.

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Almost all cell types naturally secret extracellular vesicles (EVs) in the extracellular space with variable metabolic cargo facilitating intracellular communication, posing immune-modulation capacity. Thus, "bacterial extracellular vesicles" (BEVs), with their great immunoregulatory, immune response stimulation and disease condition-altering potential, have gained importance in the medical and therapeutic industry. Various subtypes of BEVs were observed and reported in the literature, such as exosomes (30-150 nm), microvesicles (100-1000 nm), apoptotic bodies (1000-5000 nm), and oncosomes (1000-10,000 nm). As biological systems are complex entities, inserting BEVs requires extra high purity. Various techniques for BEV isolation have been employed alone or with other strategies, such as ultracentrifugation, precipitation, size-exclusion chromatography, affinity-based separation, ultrafiltration, and field-flow fractionation. But to date, no BEV isolation method is considered perfect as the lack of standard protocols limits their scale-up. Medical research has focused on BEVs to explore their diverse therapeutic potential. This review particularly focused on the recent advancements in the potential medical application of BEVs, current challenges, and prospects associated with their scale-up.

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A highly enantioselective protocol for the conjugate addition of 2-arylimidazo[1,2-a]pyridines and other imidazo derivatives to α,ß-unsaturated 2-acylimidazoles is described. The method uses a previously reported chiral-at-metal rhodium catalyst and provides the corresponding adducts in yields of 25-98% with enantioselectivities up to er > 99:1. Additionally, the transformation proceeds under mild conditions using ethanol as the solvent at room temperature.

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Herein, we report a KIO3-catalyzed oxidative coupling of thiols to their corresponding disulfides in water, in a short time and at ambient temperature. The reaction has a broad scope and exhibits good functional group tolerance, resulting in the desired products in excellent yields. This approach allows the reuse of the reaction system in multiple cycles and scale-up. Furthermore, the current protocol demonstrates compatibility for in situ generation of disulfides and post application in C(sp2)-H bond sulfenylation.

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Here we introduce a first-in-class microRNA-sensitive oncolytic Zika virus (ZIKV) for virotherapy application against central nervous system (CNS) tumors. The described methodology produced two synthetic modified ZIKV strains that are safe in normal cells, including neural stem cells, while preserving brain tropism and oncolytic effects in tumor cells. The microRNA-sensitive ZIKV introduces genetic modifications in two different virus sites: first, in the established 3'UTR region, and secondly, in the ZIKV protein coding sequence, demonstrating for the first time that the miRNA inhibition systems can be functional outside the UTR RNA sites. The total tumor remission in mice bearing human CNS tumors, including metastatic tumor growth, after intraventricular and systemic modified ZIKV administration, confirms the promise of this virotherapy as a novel agent against brain tumors-highly deadly diseases in urgent need of effective advanced therapies.

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This study describes the production, characterization and application of an endoglucanase from Penicillium roqueforti using lignocellulosic agro-industrial wastes as the substrate during solid-state fermentation. The endoglucanase was generated after culturing with different agro-industrial wastes for 96 h without any pretreatment. The highest activity was obtained at 50 °C and pH 4.0. Additionally, the enzyme showed stability in the temperature and pH ranges of 40-80 °C and 4.0-5.0, respectively. The addition of Ca2+, Zn2+, Mg2+, and Cu2+ increased enzymatic activity. Halotolerance as a characteristic of the enzyme was confirmed when its activity increased by 35% on addition of 2 M NaCl. The endoglucanase saccharified sugarcane bagasse, coconut shell, wheat bran, cocoa fruit shell, and cocoa seed husk. The Box-Behnken design was employed to optimize fermentable sugar production by evaluating the following parameters: time, substrate, and enzyme concentration. Under ideal conditions, 253.19 mg/g of fermentable sugars were obtained following the saccharification of wheat bran, which is 41.5 times higher than that obtained without optimizing. This study presents a thermostable, halotolerant endoglucanase that is resistant to metal ions and organic solvents with the potential to be applied in producing fermentable sugars for manufacturing biofuels from agro-industrial wastes.

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In this paper, we report an eco-friendly approach for the C(sp2)-H bond selenylation of imidazopyridines and other N-heteroarenes as well as simple arenes at ambient temperature. This new protocol consists of the reaction between (N-hetero)-arenes and the diorganyl-diselenides and trichloroisocyanuric acid (TCCA)-ethanol reagent system. In a short reaction time, the desired selenylated products were obtained regioselectively in good yields, with tolerance for a wide range of functional groups.

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An original total synthesis of the antiepileptic drug brivaracetam (BRV) is reported. The key step in the synthesis consists of an enantioselective photochemical Giese addition, promoted by visible-light and the chiral bifunctional photocatalyst Δ-RhS. Continuous flow conditions were employed to improve the efficiency and allow an easy scale-up of the enantioselective photochemical reaction step. The intermediate obtained from the photochemical step was converted into BRV by two different pathways, followed by one alkylation and amidation, thus giving the desired active pharmaceutical ingredients (API) in 44% overall yield, 9:1 diastereoisomeric ratio (dr) and >99:1 enantiomeric ratio (er).

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The objective was to optimize the phenolic compounds extraction from cocoa shells using the simplex-centroid design with a mixture of solvents (water, methanol, and acetone) as its components, to prove the presence of these compounds and antioxidant activity. Also, the development of dairy products, such as milk beverages and dairy desserts, with bioactive compounds, through the replacement of cocoa powder by cocoa shell was studied and evaluated sensorially. The extraction optimization indicated that a solvent with 56.44% water, 23.77% methanol, and 19.80% acetone are ideal for maximizing the phenolic compounds. In addition, the cocoa shell showed a high antioxidant activity by the methods ß-carotene/linoleic acid, FRAP, and phosphomolybdenum complex. The Check-All-That-Apply, Cochran's Q test, contingency analysis, and hierarchical cluster analysis allowed description characteristics of the dairy products and showed sensory differences between formulations with 100% cocoa shell and others. Both dairy products had good sensory acceptance in all attributes evaluated (appearance, flavor, texture, and overall impression), and their scores did not differ statistically by Tukey's test (p > 0.05). Thus, the cocoa shell is shown as an alternative substitute ingredient to be used in the dairy industry.

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This study aims at the synthesis of hexyl butyrate by Candida rugosa lipase (CRL) immobilized on Diaion HP 20. The lipase load used was 28.7 ± 2.1 mg/g (mg of lipase/g of support), whose hydrolytic activity was 132.0 ± 2.5 U/g. To obtain the maximum production of hexyl butyrate, the Box-Behnken design statistical planning was used, having as independent variables; biocatalyst concentration, temperature and acid:alcohol molar ratio and ester conversion as a dependent variable at 60, 180 and 480 min. For 60 min, 90.8% conversion was obtained at 47.25 ºC, 1:1.4 molar ratio and 17.65% of biocatalyst; 180 min, 94.5% conversion at 59.5 ºC, 1:2 molar ratio and 15.8% biocatalyst; 480 min, 95.01% conversion at 47.0 ºC, 1:2 molar ratio and 16.9% biocatalyst. CRL-Diaion HP 20 retained 60% of its initial activity after ten cycles of reactions showing potential for industrial use. The ester produced was identified by gas chromatography analyses. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01200-1.

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Ulcerative Colitis (UC) is an inflammatory disease characterized by colonic mucosal lesions associated with an increased risk of carcinogenesis. UC pathogenesis involves environmental and genetic factors. Genetic studies have indicated the association of gene variants coding for the divalent metal ion transporter SLC11A1 protein (formerly NRAMP1) with UC susceptibility in several animal species. Two mouse lines were genetically selected for high (AIRmax) or low (AIRmin) acute inflammatory responses (AIR). AIRmax is susceptible, and AIRmin is resistant to DSS-induced colitis and colon carcinogenesis. Furthermore, AIRmin mice present polymorphism of the Slc11a1 gene. Here we investigated the possible modulating effect of the Slc11a1 R and S variants in DSS-induced colitis by using AIRmin mice homozygous for Slc11a1 R (AIRminRR) or S (AIRminSS) alleles. We evaluated UC by the disease activity index (DAI), considering weight loss, diarrhea, blood in the anus or feces, cytokines, histopathology, and cell populations in the distal colon epithelium. AIRminSS mice have become susceptible to DSS effects, with higher DAI, IL6, G-CSF, and MCP-1 production and morphological and colon histopathological alterations than AIRminRR mice. The results point to a role of the Slc11a1 S allele in DSS colitis induction in the genetic background of AIRmin mice.

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Lignocellulosic residues, such as cocoa bean shell (FI), are generated in large quantities during agro-industrial activities. Proper management of residual biomass through solid state fermentation (SSF) can be effective in obtaining value-added products. The hypothesis of the present work is that the bioprocess promoted by P. roqueforti can lead to structural changes in the fibers of the fermented cocoa bean shell (FF) that confer characteristics of industrial interest. To unveil such changes, the techniques of FTIR, SEM, XRD, TGA/TG were used. After SSF, an increase of 36.6% in the crystallinity index was observed, reflecting the reduction of amorphous components such as lignin in the FI residue. Furthermore, an increase in porosity was observed through the reduction of the 2θ angle, which gives the FF a potential candidate for applications of porous products. The FTIR results confirm the reduction in hemicellulose content after SSF. The thermal and thermogravimetric tests showed an increase in the hydrophilicity and thermal stability of FF (15% decomposition) in relation to the by-product FI (40% decomposition). These data provided important information regarding changes in the crystallinity of the residue, existing functional groups and changes in degradation temperatures.

This work presents a new approach for solid state fermentation based on the study of structural changes caused by Penicillium roquefort, which is important to understand the changes in the lignocellulosic matrix after the fungus growth. The results provided important information regarding changes in the crystallinity of the residue, existing functional groups and changes in degradation temperatures. Consequently, they can help in proposals for the total use of the residual solid after fermentation, as well as contribute to reducing the lack of this information in the literature.

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The enormous amount of agroindustrial residues generated in Brazil can be used as biomass to produce fermentable sugars. This study compared the pretreatments with different proportions of dilute acid. The method involved pretreatment with 0.5%, 1%, and 1.5% (v/v) sulfuric acid, followed by hydrolysis using the halotolerant and thermostable endoglucanase from Botrytis ricini URM 5627. The physicochemical characterization of plant biomass was performed using XRD, FTIR, and SEM. The pretreatment significantly increased the production of fermentable sugars following enzymatic saccharification from wheat bran, sugarcane bagasse, and rice husk: 153.67%, 91.98%, and 253.21% increment in sugar production; 36.39 mg⋅g-1 ± 1.23, 39.55 mg⋅g-1 ± 1.70, and 42.53 mg⋅g-1 ± 7.61 mg⋅L-1 of glucose; and 3.26 ± 0.35 mg⋅g-1 , 3.61mg⋅g-1 ± 0.74 and 3.59 mg⋅g-1 ± 0.80 of fructose were produced, respectively. In conclusion, biomass should preferably be pretreated before the enzymatic saccharification using B. ricini URM 5627 endoglucanase.

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Guava juice is cloudy and viscous, which hinders filtration, decreases yield, and causes the loss of quality after its processing and during storage. This study aimed to evaluate enzymatic treatment effects using crude multi-enzymatic extracts (CME) obtained from Rhodotorula mucilaginosa, Rhodotorula orizycola, and Pseudozyma sp. produced by submerse fermentation in the extraction of juice guava. Mixtures of 100 ml of guava pulp and multi-enzymatic extracts proposed by Doehlert planning were incubated under constant agitation at 150 rpm and 50°C, and a Doehlert design was applied as a multivariate optimization strategy. The optimal conditions using the multi-enzymatic extract were: 0.4% (v/v) of CME for 131 min for the multi-enzymatic treatment using Pseudozyma sp.; 3.0% (v/v) of CME for 154 min using the R. mucilaginosa CME; and 5.0% (v/v) of CME for 90 min using R. oryzicola. The maximum viscosity reduction values for the juices treated with the CME of yeasts were 10.33%, 86.38%, and 13.33% for the juices treated with the CME of Pseudozyma sp., R. mucilaginosa, and R. orizycola, respectively. The physical-chemical properties were improved after treatment with CMEs, yielding a reduction of clarity, increase of total soluble solids and reducing sugars, and decreasing the acidity (pH) for all treatments with enzymatic extracts of all strains. The yeasts studied showed a potential for CME production to be applied to juice, improving the quality of the juice, and R. mucilaginosa was the most prominent yeast due to most significant reduction of viscosity in guava juice.

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Ulcerative Colitis (UC) is an inflammatory disease characterized by colonic mucosal lesions associated with an increased risk of carcinogenesis. UC pathogenesis involves environmental and genetic factors. Genetic studies have indicated the association of gene variants coding for the divalent metal ion transporter SLC11A1 protein (formerly NRAMP1) with UC susceptibility in several animal species. Two mouse lines were genetically selected for high (AIRmax) or low (AIRmin) acute inflammatory responses (AIR). AIRmax is susceptible, and AIRmin is resistant to DSS-induced colitis and colon carcinogenesis. Furthermore, AIRmin mice present polymorphism of the Slc11a1 gene. Here we investigated the possible modulating effect of the Slc11a1 R and S variants in DSS-induced colitis by using AIRmin mice homozygous for Slc11a1 R (AIRminRR) or S (AIRminSS) alleles. We evaluated UC by the disease activity index (DAI), considering weight loss, diarrhea, blood in the anus or feces, cytokines, histopathology, and cell populations in the distal colon epithelium. AIRminSS mice have become susceptible to DSS effects, with higher DAI, IL6, G-CSF, and MCP-1 production and morphological and colon histopathological alterations than AIRminRR mice. The results point to a role of the Slc11a1 S allele in DSS colitis induction in the genetic background of AIRmin mice.

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We report herein the synthesis of primary and secondary ß-chalcogen amines through the regioselective ring-opening reaction of non-activated 2-oxazolidinones promoted by in situ generated chalcogenolate anions. The developed one-step protocol enabled the preparation of ß-selenoamines, ß-telluroamines and ß-thioamines with appreciable structural diversity and in yields of up to 95%.

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Resumo O tema segurança tem sido intensamente discutido, mostrando-se cada vez mais relevante na saúde pública e em projetos de pesquisa envolvendo seres humanos. Participantes de estudos clínicos estão sujeitos a riscos, físicos ou não, que impactam em sua integridade, direitos ou autonomia. Este trabalho apresenta e discute a atuação do Comitê de Monitoramento de Dados e de Segurança para a proteção do participante de pesquisa e minimização de riscos em pesquisa clínica. A metodologia consiste em revisão integrativa da literatura, realizada com o propósito de identificar as funções dos comitês e seu papel na proteção dos participantes. Identificou-se que grande parte das publicações analisadas confirmam que os comitês de monitoramento têm como responsabilidade principal a proteção do participante de pesquisa, além da garantia de integridade e credibilidade da pesquisa.

Abstract Of increasingly relevance in public health and research projects involving human beings, the topic of safety has been intensely discussed. Participants in clinical trials are subject to risks, physical or otherwise, that impact their integrity, rights, or autonomy. This study outlines and discusses the performance of the Data and Safety Monitoring Committee for research participant protection and risk minimization in clinical research. An integrative literature review was conducted to identify the committees' duties and role in protecting participants. Most of the analyzed articles confirm that the monitoring committees are mainly responsible for protecting research participants, as well as ensuring research integrity and credibility.

Resumen La seguridad ha sido un tema muy discutido, por lo que muestra su relevancia para la salud pública y los proyectos de investigación que involucran a seres humanos. Los participantes en estudios clínicos están sujetos a riesgos físicos o de otro tipo, que impactarán su integridad, derechos o autonomía. Este texto realiza un debate sobre el desempeño del Comité de Seguimiento de Datos y Seguridad destinado a la protección de los participantes de investigación y la mitigación de los riesgos en investigación clínica. Se realizó una revisión integradora de la literatura, con el propósito de identificar las funciones de los comités y su rol en la protección de los participantes. La mayoría de las publicaciones analizadas confirman que los comités de seguimiento tienen como principal responsabilidad la protección del participante de la investigación, además de garantizar la integridad y credibilidad de la investigación.

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We identified Pycard and BC017158 genes as putative effectors of the Quantitative Trait locus (QTL) that we mapped at distal chromosome 7 named Irm1 for Inflammatory response modulator 1, controlling acute inflammatory response (AIR) and the production of IL-1ß, dependent on the activation of the NLRP3 inflammasome. We obtained the mapping through genome-wide linkage analysis of Single Nucleotide Polymorphisms (SNPs) in a cross between High (AIRmax) and Low (AIRmin) responder mouse lines that we produced by several generations of bidirectional selection for Acute Inflammatory Response. A highly significant linkage signal (LOD score peak of 72) for ex vivo IL-1ß production limited a 4 Mbp interval to chromosome 7. Sequencing of the locus region revealed 14 SNPs between "High" and "Low" responders that narrowed the locus to a 420 Kb interval. Variants were detected in non-coding regions of Itgam, Rgs10 and BC017158 genes and at the first exon of Pycard gene, resulting in an E19K substitution in the protein ASC (apoptosis associated speck-like protein containing a CARD) an adaptor molecule in the inflammasome complex. Silencing of BC017158 inhibited IL1-ß production by stimulated macrophages and the E19K ASC mutation carried by AIRmin mice impaired the ex vivo IL-1ß response and the formation of ASC specks in stimulated cells. IL-1ß and ASC specks play major roles in inflammatory reactions and in inflammation-related diseases. Our results delineate a novel genetic factor and a molecular mechanism affecting the acute inflammatory response.

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Lipases (triacylglycerol hydrolases, EC 3.1.1.3) are a class of enzymes with high industrial importance. An option for the production of this enzyme is through fungal growth via solid-state fermentation (SSF). Thus, this research presents a study of lipase production by Penicillium roqueforti ATCC 10110 through SSF using cocoa bran residues (Theobroma cacao) as a substrate. To achieve maximum lipase production, fermentation time (0 to 120 h) and palm oil (PO) percentage (0 to 50%) were optimized through analysis of one factor at a time (OFAT), with lipase activity as the response. The amount of cocoa was fixed (5 g), the incubation temperature was maintained at 27 °C, and the moisture content was established at 70%. For a 72 h incubation, the highest enzyme activity achieved using SSF without adding PO was 14.67 ± 1.47 U g-1, whereas with PO (30%), it was 33.33 ± 3.33 U g-1, thus demonstrating a 44% increase in enzyme activity. Through the OFAT methodology, it was possible to confirm that supplementation with palm residue was efficient and maximized the lipase of P. roqueforti ATCC 10110.

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