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Two artisanal fermentation processes for Criollo cocoa beans with different turning start times (24 h and 48 h) were studied. The aromatic profile of cocoa turned every 24 h (B1) displayed volatile compounds associated with fermented, bready, and fruity aromas. When cocoa beans were fermented with a different turning technique with a start time of 48 h (B2), they provided volatile compounds mainly associated with descriptors of floral, woody, sweet, fruity and chocolate aromas. The turning start time of 48 h stimulated a microbial profile dominated by yeast such as Hanseniaspora opuntiae, Pichia manshurica, and Meyerozyma carpophila, favoring the production of several key aroma markers associated with cocoa bean fermentation quality, such as phenylethyl acetate, 2-phenylacetaldehyde, 3-methylbutanal, 2-phenylethyl alcohol, 2,3-butanedione, 3-methylbutanoic acid, and 2-methylpropanoic acid, while an immediate turning start time (24 h) favored an aerobic environment that stimulated the rapid growth of Acetobacter pasteurianus, Bacillus subtilis and a higher biodiversity of lactic acid bacteria (LAB) (e.g., Lactobacillus plantarum and Pediococcus acidilactici), which increased the production of ethyl acetate and 3-hydroxy-2-butanone. Volatile compound generation and microbial populations were evaluated and analyzed by multivariate analysis (principal component analysis and partial least squares discriminant analysis) to find correlations and significant differences. This study shows that the method of turning Criollo cacao beans can lead to the formation of desirable aromatic compounds.

Bacillus Calmette-Guèrin (BCG) remains as the only vaccine employed to prevent tuberculosis (TB) during childhood. Among factors likely contributing to the variable efficacy of BCG is the modification in its antigenic repertoire that may arise from in vitro growth conditions. Our vaccine candidate, BCGΔBCG1419c, improved protection against TB in mice and guinea pigs with bacteria grown in either 7H9 OADC Tween 80 or in Proskauer Beck Tween 80 media in independent studies. Here, we compared the proteomes of planktonic cultures of BCG and BCGΔBCG1419c, grown in both media. Further to this, we compared systemic immunogenicity ex vivo elicited by both types of BCG strains and cultures when used to vaccinate BALB/c mice. Both the parental strain BCG Pasteur ATCC 35734, and its isogenic mutant BCGΔBCG1419c, had several medium-dependent changes. Moreover, ex vivo immune responses to a multiantigenic (PPD) or a single antigenic (Ag85A) stimulus were also medium-dependent. Then, not only the presence or absence of the BCG1419c gene in our strains under study affected the proteome produced in vitro but also that this was affected by culture medium, potentially leading to changes in the capacity to induce ex vivo immune responses., Competing Interests: Declaration of competing interest M.A.F.V., C.P.R., and M.J.A.S are inventors in the patent 363576 issued in Mexico for the BCGΔBG1419c vaccine candidate. All other authors have no interests to declare., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Cocoa bean fermentation is an important process because during this process, aroma compounds are produced, the astringency decreases, and the embryo dies. The fermentation processes of the Criollo and Forastero types have been studied separately without comparing them at the same time and in the same place. The aim of this work was to determine differences in the profile of volatile and nonvolatile compounds of Criollo and Forastero cocoa from the fermentation process to the final stage of obtaining the liquor. The experiments were carried out at the same time in the Maya region. Volatile compounds were determined by HS-SPME GC-MS (headspace solid phase-microextraction with gas chromatography-mass spectrometry). Sugars, organic acids, and alkaloids were determined by ultrahigh-performance liquid chromatography (UHPLC-PDA/UV). Criollo cocoa liquor was defined by the volatile and nonvolatile compounds such as acetic acid, phenylethyl alcohol, benzaldehyde, 2-phenylethyl acetate, acetophenone and 3-methylbutanal., which are associated with sour, honey, almond, flowery and chocolate aroma. Forastero cocoa liquor was represented with a significant difference by acetic acid, isobutyl acetate, 2,3-diethyl-5-methylpyrazine and ethyl octanoate and these could provide aroma descriptors such as sour, fruity and nutty. This study characterized for the first time the dynamics of volatile compounds during the fermentation, drying, and roasting stages and in the final cocoa liquor of Criollo and Forastero from cocoa beans of the same origin., Competing Interests: The authors declare no conflict of interest., (©2023PublishedbyElsevierLtd.)

Vinasses represent important final disposal problems due to their physical-chemical composition. This work analyzed the composition of tequila vinasses and increased 5-hydroxymethylfurfural, furfural, and phenolic compounds using thermal hydrolysis with hydrogen peroxide as a catalyst. A statistical Taguchi design was used, and a UPLC-MS (XEVO TQS Micro) analysis determined the presence and increase of the components. The treatment at 130 °C, 40 min, and 0.5% of catalyst presented the highest increase for 5-HMF (127 mg/L), furfural (3.07 mg/L), and phenol compounds as chlorogenic (0.36 mg/L), and vanillic acid (2.75 mg/L). Additionally, the highest removal of total sugars (57.3%), sucrose (99.3%), and COD (32.9%). For the treatment T130:30m:0P the syringic (0.74 mg/L) and coumaric (0.013 mg/L) acids obtained the highest increase, and the treatment T120:30m:1P increased 3-hydroxybenzoic (1.30 mg/L) and sinapic (0.06 mg/L) acid. The revaluation of vinasses through thermal treatments provides guidelines to reduce the impact generated on the environment.

Carotene cleavage dioxygenases (CCDs) are a large family of Fe 2+ dependent enzymes responsible for the production of a wide variety of apocarotenoids, such as bixin. Among the natural apocarotenoids, bixin is second in economic importance. It has a red-orange color and is produced mainly in the seeds of B. orellana . The biosynthesis of bixin aldehyde from the oxidative cleavage of lycopene at 5,6/5',6' bonds by a CCD is considered the first step of bixin biosynthesis. Eight BoCCD ( BoCCD1-1, BoCCD1-3, BoCCD1-4, CCD4-1, BoCCD4-2, BoCCD4-3 and BoCCD4-4 ) genes potentially involved in the first step of B. orellana bixin biosynthesis have been identified. However, the cleavage activity upon lycopene to produce bixin aldehyde has only been demonstrated for BoCCD1-1 and BoCCD4-3. Using in vivo ( Escherichia coli ) and in vitro approaches, we determined that the other identified BoCCDs enzymes (BoCCD1-3, BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4) also participate in the biosynthesis of bixin aldehyde from lycopene. The LC-ESI-QTOF-MS/MS analysis showed a peak corresponding to bixin aldehyde ( m/z 349.1) in pACCRT-EIB E. coli cells that express the BoCCD1 and BoCCD4 proteins, which was confirmed by in vitro enzymatic assay. Interestingly, in the in vivo assay of BoCCD1-4, BoCCD4-1, BoCCD4-2, and BoCCD4-4, bixin aldehyde was oxidized to norbixin ( m/z 380.2), the second product of the bixin biosynthesis pathway. In silico analysis also showed that BoCCD1 and BoCCD4 proteins encode functional dioxygenases that can use lycopene as substrate. The production of bixin aldehyde and norbixin was corroborated based on their ion fragmentation pattern, as well as by Fourier transform infrared (FTIR) spectroscopy. This work made it possible to clarify at the same time the first and second steps of the bixin biosynthesis pathway that had not been evaluated for a long time., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Us-Camas, Aguilar-Espinosa, Rodríguez-Campos, Vallejo-Cardona, Carballo-Uicab, Serrano-Posada and Rivera-Madrid.)

Tuberculosis (TB) is the most important infectious disease worldwide, based on the number of new cases and deaths reported by the World Health Organization. Several vaccine candidates against TB have been characterized at preclinical and clinical levels. The BCGΔBCG1419c vaccine candidate, which lacks the BCG1419c gene that encodes for a c-di-GMP phosphodiesterase, provides improved efficacy against chronic TB, reactivation from latent-like infection and against chronic TB in the presence of type 2 diabetes in murine models. We previously reported that compared with wild type BCG, BCGΔBCG1419c changed levels of several proteins. Here, using a label-free proteomic approach, we confirmed that a novel, second-generation version of BCGΔBCG1419c maintains changes in antigenic proteins already reported, and here we further found differences in secreted proteins, as well as that this new BCGΔBCG1419c version modifies its production of proteins involved in redox and nitrogen/protein metabolism compared with wild type BCG. This work contributes to the proteomic characterization of a novel vaccine candidate that is more effective against TB than parental BCG in diverse murine models., (© The Author(s) 2020. Published by Oxford University Press on behalf of FEMS.)

Fermented cocoa beans can be described as a complex matrix that integrates the chemical history of beans, their processing, and environmental factors. This study presents an analysis that aims to identify volatile compounds of five varieties of fine-aroma cocoa types. The cocoa types studied were Carmelo, Rojo Samuel, Lagarto, Arcoiris, Regalo de Dios, that grow in the Maya lands of Chiapas, Mexico. Profile of volatile compounds was obtained from each cacao type during fermentation and drying process. This profile of volatile compounds also was compared with beans unfermented, using a statistical analysis of Venn diagram and a multivariate Analysis of Principal Components (PCA). One hundred nine different compounds were identified by SPME-HS GC-MS, these compounds mainly related to desirable aromatic notes generated by esters, aldehydes, ketones, and alcohols. The differences in chemical composition of the volatile compounds were associated mainly with the process and not to cocoa varieties. Fermented dry cocoa beans showed a higher content of esters, aldehydes, pyrazines, alcohols, some acids, and furans where Lagarto (CL), Rojo Samuel (CR), and Regalo de Dios (TRD) cocoas type showed a more interesting aromatic profile. On the other hand, as expected dry unfermented cocoas presented a few numbers of aroma compounds, in the five cacao types, where alcohols, ketones and hydrocarbons predominated., Competing Interests: Declaration of Competing Interest We have no conflicts of interest to disclose. The manuscript has been read and approved for submission by all the named authors. Declaration of Competing Interest The authors declare that they have no conflicts of interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Mycobacterium tuberculosis produces mycolic acids which are relevant for persistence, recalcitrance to antibiotics and defiance to host immunity. c-di-GMP is a second messenger involved in transition from planktonic cells to biofilms, whose levels are controlled by diguanylate cyclases (DGC) and phosphodiesterases (PDE). The transcriptional regulator dosR, is involved in response to low oxygen, a condition likely happening to a subset of cells within biofilms. Here, we found that in M. bovis BCG, expression of both BCG1416c and BCG1419c genes, which code for a DGC and a PDE, respectively, decreased in both stationary phase and during biofilm production. The kasA , kasB , and fas genes, which are involved in mycolic acid biosynthesis, were induced in biofilm cultures, as was dosR , therefore suggesting an inverse correlation in their expression compared with that of genes involved in c-di-GMP metabolism. The relative abundance within trehalose dimycolate (TDM) of α-mycolates decreased during biofilm maturation, with methoxy mycolates increasing over time, and keto species remaining practically stable. Moreover, addition of synthetic c-di-GMP to mid-log phase BCG cultures reduced methoxy mycolates, increased keto species and practically did not affect α-mycolates, showing a differential effect of c-di-GMP on keto- and methoxy-mycolic acid metabolism.

Asclepias linaria Cav. (Apocynaceae) is a shrubby plant endemic of Mexico which has been used in traditional medicine. However, the bioactive potential of this plant remains unexplored. In this study, the phenolic composition, antioxidant, and cytotoxic activities of A. linaria leaves were determined. In order to estimate the phenolic composition of the leaves, the total phenolic, flavonoid, and condensed tannins contents were determined. Furthermore, the antioxidant activity was measured by the scavenging activity of the 2,2-diphenyl-1-picrylhydrazyl (DPPH • ) and 2,2'-azino-bis[3-ethylbenzothiazoline-6-sulphonic acid] (ABTS •+ ) radicals and the total antioxidant capacity. The phenolic compounds identified in the A. linaria leaves by ultra-performance liquid chromatography coupled to mass spectrometry (UPLC-MS) include phenolic acids, such as p -coumaric and ferulic acid, as well as flavonoids, such as rutin and quercetin. The leaves' extracts of A. linaria showed a high scavenging activity of DPPH • and ABTS •+ radicals (IC 50 0.12 ± 0.001 and 0.51 ± 0.003 µg/mL, respectively), high total antioxidant capacity values (99.77 ± 4.32 mg of ascorbic acid equivalents/g of dry tissue), and had a cytotoxic effect against K562 and HL60 hematologic neoplasia cells lines, but no toxicity towards the normal mononuclear cell line was observed. These results highlight the potential of A. linaria and could be considered as a possible alternative source of anticancer compounds.

Carotenoid cleavage dioxygenases (CCDs) are enzymes that have been implicated in the biosynthesis of a wide diversity of secondary metabolites with important economic value, including bixin. Bixin is the second most used pigment in the world's food industry worldwide, and its main source is the aril of achiote ( Bixa orellana L.) seeds. A recent transcriptome analysis of B. orellana identified a new set of eight CCD members (BoCCD4s and BoCCD1s) potentially involved in bixin synthesis. We used several approaches in order to discriminate the best candidates with CCDs genes. A reverse transcription-PCR (RT-qPCR) expression analysis was carried out in five developmental stages of two accessions of B. orellana seeds with different bixin contents: (P13W, low bixin producer and N4P, high bixin producer). The results showed that three BoCCDs (BoCCD4-1, BoCCD4-3, and BoCCD1-1) had an expression pattern consistent with bixin accumulation during seed development. Additionally, an alignment of the CCD enzyme family and homology models of proteins were generated to verify whether the newly proposed CCD enzymes were bona fide CCDs. The study confirmed that these three enzymes were well-preserved and belonged to the CCD family. In a second selection round, the three CCD genes were analyzed by in situ RT-qPCR in seed tissue. Results indicated that BoCCD4-3 and BoCCD1-1 exhibited tissue-specific expressions in the seed aril. To test whether the two selected CCDs had enzymatic activity, they were expressed in Escherichia coli ; activity was determined by identifying their products in the crude extract using UHPLC-ESI-QTOF-MS/MS. The cleavage product (bixin aldehyde) was also analyzed by Fourier transform infrared. The results indicated that both BoCCD4-3 and BoCCD1-1 cleave lycopene in vitro at 5,6-5',6'., Competing Interests: The authors declare that they have no competing interests.

Type A Aortic Dissection is a medical-surgical emergency which requires prompt diagnosis and adequate treatment. Since its inception--more than 40 years ago--the surgical treatment has evolved up until now, when it offers an immediate solution to the high mortality rate due to complications, with acceptable morbidity and mortality rates. This improvement is due also to better diagnostic techniques, postoperative management and profound hypothermia with circulatory arrest. The basic techniques to achieve this consist of resection of the entry tear and closure of the false lumen and repair of the aortic regurgitation when present. Failure of them--proximally or distally--leads to persistence of the false lumen and the possibility of complications or late reoperations. The long-term follow-up, monitorized by non-invasive methods, is mandatory to decide the proper management.