Optimization of HS-SPME/GC-MS Method for Determining Volatile Organic Compounds and Sensory Profile in Cocoa Honey from Different Cocoa Varieties (Theobroma cacao L.).

This study aimed to develop an analytical method using HS-SPME/GC-MS to determine the volatile organic compound (VOC) profiles and evaluate the sensory attributes of cocoa honey from four cocoa varieties (CCN51, PS1319, SJ02, and Parazinho). Using a multivariate factorial experimental design, the HS-SPME/GC-MS method was optimized to determine the VOC profiles. Twenty previously trained tasters participated in the ranking descriptive analysis, while 108 consumers participated in the acceptance and purchase intention tests. A total of 84 volatile organic compounds were identified from various chemical classes, including acids, alcohols, aldehydes, esters, ketones, monoterpenes, oxygenated monoterpenoids, sesquiterpenes, and oxygenated sesquiterpenoids. Palmitic acid was the compound found in the highest concentration in all varieties (5.13-13.10%). Multivariate analysis tools identified key compounds for differentiation and grouping of the samples. The results revealed that the variety significantly influenced both the VOCs' concentrations and sensory profiles. The CCN51, PS1319, and SJ02 varieties exhibited the highest diversity of VOCs and sensory attributes. Notably, the SJ02 and CCN51 varieties demonstrated superior acceptability and purchase intention, with means ranging from 7.21 and 7.08 to 3.71 and 3.56, respectively. These results indicate their potential as promising sources of cocoa honey for the food industry.

Effects of Infrared and Microwave Radiation on the Bioactive Compounds of Microalga Spirulina platensis after Continuous and Intermittent Drying.

Pharmaceutical, nutritional and food industries have recently become interested in the potential of Spirulina platensis, a kind of cyanobacterium with high levels of proteins, vitamins and bioactive compounds. Because of its high moisture, this microalga needs to be submitted to a preservation technique such as drying to be properly used. The aim of this work is to investigate the use of infrared and microwave radiation in the Spirulina platensis drying process. The experiments were performed in continuous and intermittent modes, evaluating different operating conditions for infrared and microwave drying, as well as their effects on the quality of the final product, expressed by the content of bioactive compounds (i.e., total phenolic, total flavonoid, citric acid and phycocyanin contents). The results proved that the use of electromagnetic radiation in the drying of spirulina is an interesting alternative for processing this material if performed under adequate operating conditions. The experiments carried out continuously at lower temperatures and powers and the combination between different temperatures and powers in the intermittent mode resulted in a final product with satisfactory levels of bioactive compounds and low operation times in comparison with conventional methodologies.

Revisiting the Spectrum of Co(CN)63-: The Role of Correlation, Solvation, and Vibronic and Spin-Orbit Couplings.

In the present work, we revisit the spectrum of the hexacyanocobaltate(III) ion, [Co(CN)6]3-, which has been considered a prototype complex in the coordination chemistry, with modern quantum chemistry methods. The main features have been describing by revealing the role of different effects, such as vibronic coupling, solvation and spin-orbit coupling. The UV-vis spectrum is composed by two bands (1A1g → 1T1g and 1A1g → 1T2g), characterized by singlet-singlet metal-centered transitions, and a more intense third one, characterized by charge transfer transition. There is also a small band shoulder. The first two are symmetry-forbidden transitions in the Oh group. Their intensity can only be explained by a vibronic coupling mechanism. For the band shoulder, additional to vibronic coupling, spin-orbit coupling is also necessary, since the transition is characterized as singlet to triplet, 1A1g → 3T1g.

Single-cell transcriptomic analysis of endometriosis.

Endometriosis is a common condition in women that causes chronic pain and infertility and is associated with an elevated risk of ovarian cancer. We profiled transcriptomes of >370,000 individual cells from endometriomas (n = 8), endometriosis (n = 28), eutopic endometrium (n = 10), unaffected ovary (n = 4) and endometriosis-free peritoneum (n = 4), generating a cellular atlas of endometrial-type epithelial cells, stromal cells and microenvironmental cell populations across tissue sites. Cellular and molecular signatures of endometrial-type epithelium and stroma differed across tissue types, suggesting a role for cellular restructuring and transcriptional reprogramming in the disease. Epithelium, stroma and proximal mesothelial cells of endometriomas showed dysregulation of pro-inflammatory pathways and upregulation of complement proteins. Somatic ARID1A mutation in epithelial cells was associated with upregulation of pro-angiogenic and pro-lymphangiogenic factors and remodeling of the endothelial cell compartment, with enrichment of lymphatic endothelial cells. Finally, signatures of ciliated epithelial cells were enriched in ovarian cancers, reinforcing epidemiologic associations between these two diseases.

Use of Refractance Window Drying as an Alternative Method for Processing the Microalga Spirulina platensis.

Microalgae such as Spirulina platensis have recently attracted the interest of the pharmaceutical, nutritional and food industries due to their high levels of proteins and bioactive compounds. In this study, we investigated the use of refractance window (RW) drying as an alternative technology for processing the microalga Spirulina biomass aiming at its dehydration. In addition, we also analyzed the effects of operating variables (i.e., time and temperature) on the quality of the final product, expressed by the content of bioactive compounds (i.e., total phenolics, total flavonoids, and phycocyanin). The results showed that RW drying can generate a dehydrated product with a moisture content lower than 10.0%, minimal visual changes, and reduced process time. The content of bioactive compounds after RW drying was found to be satisfactory, with some of them close to those observed in the fresh microalga. The best results for total phenolic (TPC) and total flavonoids (TFC) content were obtained at temperatures of around 70 °C and processing times around 4.5 h. The phycocyanin content was negatively influenced by higher temperatures (higher than 80 °C) and high exposing drying times (higher than 4.5 h) due to its thermosensibility properties. The use of refractance window drying proved to be an interesting methodology for the processing and conservation of Spirulina platensis, as well as an important alternative to the industrial processing of this biomass.

Epigenomic charting and functional annotation of risk loci in renal cell carcinoma.

While the mutational and transcriptional landscapes of renal cell carcinoma (RCC) are well-known, the epigenome is poorly understood. We characterize the epigenome of clear cell (ccRCC), papillary (pRCC), and chromophobe RCC (chRCC) by using ChIP-seq, ATAC-Seq, RNA-seq, and SNP arrays. We integrate 153 individual data sets from 42 patients and nominate 50 histology-specific master transcription factors (MTF) to define RCC histologic subtypes, including EPAS1 and ETS-1 in ccRCC, HNF1B in pRCC, and FOXI1 in chRCC. We confirm histology-specific MTFs via immunohistochemistry including a ccRCC-specific TF, BHLHE41. FOXI1 overexpression with knock-down of EPAS1 in the 786-O ccRCC cell line induces transcriptional upregulation of chRCC-specific genes, TFCP2L1, ATP6V0D2, KIT, and INSRR, implicating FOXI1 as a MTF for chRCC. Integrating RCC GWAS risk SNPs with H3K27ac ChIP-seq and ATAC-seq data reveals that risk-variants are significantly enriched in allelically-imbalanced peaks. This epigenomic atlas in primary human samples provides a resource for future investigation.

Bidirectional associations between chronic low back pain and sleep quality: A cohort study with schoolteachers.

AIMS: Although both chronic low back pain (cLBP) and sleep problems are prevalent among active workers, the relation between these variables is not well established. This study aimed to examine the bidirectional association between cLBP and sleep in schoolteachers. METHODS: The Pittsburgh Sleep Quality Index (PSQI) and cLBP were self-reported by 530 schoolteachers in Londrina, Brazil, at baseline and after 2 years of follow-up. Generalized estimating equations were adjusted for sociodemographic, lifestyle and mental health variables. RESULTS: Poor sleep quality at baseline was associated with cLBP at follow-up after adjusting for sociodemographic and lifestyle variables (OR=1.61; 95% confidence interval [95% CI]=1.06, 2.47). Changes in the PSQI score over time were also associated with a higher likelihood of cLBP at follow-up (OR=1.13; 95% CI=1.07, 1.20 for each 1-point increase in the PSQI score), regardless of mental health condition. cLBP at baseline was associated with worse sleep quality at follow-up after adjusting for sociodemographic and lifestyle variables (OR=1.56; 95% CI=1.02, 2.37). The presence of cLBP also changed the PSQI score over time (ß coefficient=1.153; 95% CI=0.493, 1.814). CONCLUSIONS: Worse sleep quality was prospectively and bidirectionally associated with cLBP. Concretely, changes in PSQI values after 2 years of follow-up increased the likelihood of reporting cLBP, and baseline cLBP was associated with sleep quality worsening (i.e., higher score in the PSQI). Mental health conditions such as self-rated health, depression and anxiety play a relevant confounding role in the bidirectional associations between sleep and chronic low back pain.

Hydroethanolic Extract of Grape Peel from Vitis labrusca Winemaking Waste: Antinociceptive and Anti-Inflammatory Activities.

Research background: Extracts from grape pomace, including the wine, show many biological effects such as antioxidant and anti-inflammatory activities. Unfortunately, winemakers discard the bagasse, so the waste is not exploited, although it contains bioactive compounds with antioxidant and anti-inflammatory properties. The work aims to analyze the hydroethanolic extract of peels from Vitis labrusca agro-industrial waste and to evaluate its antinociceptive and anti-inflammatory properties. This study is relevant for reusing a residue and adding value to the grape economic chain. Experimental approach: A representative sample of pomace was obtained and the peels were used to produce the extract. The phenolic compounds were determined by mass spectrometry in multiple reaction monitoring mode and Folin-Ciocalteu colorimetric method, using gallic acid as standard. The biological analyses were carried out using mice orally treated with crude extract at doses of 30, 100 and 300 mg/kg. We evaluated mechanical hyperalgesia by the von Frey method, thermal heat hyperalgesia using a hot plate at 55 °C, paw edema using a pachymeter, and neutrophil recruitment by measurement of myeloperoxidase activity. The nephrotoxicity and hepatotoxicity were evaluated by biochemical analyses using blood samples that were collected after the Vitis labrusca administration. Results and conclusions: In all wet winemaking residues peel mass fraction was 75%, and in dry residues 59%. We identified nine anthocyanins (3-O-glucosides: peonidin, delphinidin, petunidin and malvidin; 3-p-coumaroyl-glucosides: cyanidin, peonidin, petunidin and malvidin, and malvidin-3,5-diglucoside), five flavonoids (apigenin-7-glucoside, luteolin-7-glucoside, quercetin-3-galactoside, isorhamnetin-3-glucoside and myricetin-3-rutinoside), and mass fraction of phenolic compounds, expressed as gallic acid equivalents, was 26.62 mg/g. In vivo assays showed that Vitis labrusca extract at mass fractions 100 and 300 mg/kg reduced carrageenan-induced mechanical and thermal hyperalgesia, 50% of the paw edema, and neutrophil recruitment. In addition, there were no indications of nephrotoxicity and hepatotoxicity. Our extract obtained from winemaking residue has analgesic and anti-inflammatory properties, related at least in part to the presence of phenolic compounds, and it is not toxic to renal and hepatic tissues. Novelty and scientific contribution: This bio-product can be used as an alternative to synthetic anti-inflammatory agents with the same pharmacological potential and fewer side effects. We demonstrated that Vitis labrusca winemaking waste can be used for the production of antinociceptive and anti-inflammatory products (nutraceutical, pharmaceutical and cosmetics) without toxicity, contributing to the environmental economy.

Predicting master transcription factors from pan-cancer expression data.

Critical developmental "master transcription factors" (MTFs) can be subverted during tumorigenesis to control oncogenic transcriptional programs. Current approaches to identifying MTFs rely on ChIP-seq data, which is unavailable for many cancers. We developed the CaCTS (Cancer Core Transcription factor Specificity) algorithm to prioritize candidate MTFs using pan-cancer RNA sequencing data. CaCTS identified candidate MTFs across 34 tumor types and 140 subtypes including predictions for cancer types/subtypes for which MTFs are unknown, including e.g. PAX8, SOX17, and MECOM as candidates in ovarian cancer (OvCa). In OvCa cells, consistent with known MTF properties, these factors are required for viability, lie proximal to superenhancers, co-occupy regulatory elements globally, co-bind loci encoding OvCa biomarkers, and are sensitive to pharmacologic inhibition of transcription. Our predictions of MTFs, especially for tumor types with limited understanding of transcriptional drivers, pave the way to therapeutic targeting of MTFs in a broad spectrum of cancers.

Taming the Antiferromagnetic Beast: Computational Design of Ultrashort Mn-Mn Bonds Stabilized by N-Heterocyclic Carbenes.

The development of complexes featuring low-valent, multiply bonded metal centers is an exciting field with several potential applications. In this work, we describe the design principles and extensive computational investigation of new organometallic platforms featuring the elusive manganese-manganese bond stabilized by experimentally realized N-heterocyclic carbenes (NHCs). By using DFT computations benchmarked against multireference calculations, as well as MO- and VB-based bonding analyses, we could disentangle the various electronic and structural effects contributing to the thermodynamic and kinetic stability, as well as the experimental feasibility, of the systems. In particular, we explored the nature of the metal-carbene interaction and the role of the ancillary η6 coordination to the generation of Mn2 systems featuring ultrashort metal-metal bonds, closed-shell singlet multiplicities, and positive adiabatic singlet-triplet gaps. Our analysis identifies two distinct classes of viable synthetic targets, whose electrostructural properties are thoroughly investigated.

In vivo discovery of RNA proximal proteins via proximity-dependent biotinylation.

RNA molecules function as messenger RNAs (mRNAs) that encode proteins and noncoding transcripts that serve as adaptor molecules, structural components, and regulators of genome organization and gene expression. Their function and regulation are largely mediated by RNA binding proteins (RBPs). Here we present RNA proximity labelling (RPL), an RNA-centric method comprising the endonuclease-deficient Type VI CRISPR-Cas protein dCas13b fused to engineered ascorbate peroxidase APEX2. RPL discovers target RNA proximal proteins in vivo via proximity-based biotinylation. RPL applied to U1 identified proteins involved in both U1 canonical and noncanonical functions. Profiling of poly(A) tail proximal proteins uncovered expected categories of RBPs and provided additional evidence for 5'-3' proximity and unexplored subcellular localizations of poly(A)+ RNA. Our results suggest that RPL allows rapid identification of target RNA binding proteins in native cellular contexts, and is expected to pave the way for discovery of novel RNA-protein interactions important for health and disease.

Reprogramming of the FOXA1 cistrome in treatment-emergent neuroendocrine prostate cancer.

Lineage plasticity, the ability of a cell to alter its identity, is an increasingly common mechanism of adaptive resistance to targeted therapy in cancer. An archetypal example is the development of neuroendocrine prostate cancer (NEPC) after treatment of prostate adenocarcinoma (PRAD) with inhibitors of androgen signaling. NEPC is an aggressive variant of prostate cancer that aberrantly expresses genes characteristic of neuroendocrine (NE) tissues and no longer depends on androgens. Here, we investigate the epigenomic basis of this resistance mechanism by profiling histone modifications in NEPC and PRAD patient-derived xenografts (PDXs) using chromatin immunoprecipitation and sequencing (ChIP-seq). We identify a vast network of cis-regulatory elements (N~15,000) that are recurrently activated in NEPC. The FOXA1 transcription factor (TF), which pioneers androgen receptor (AR) chromatin binding in the prostate epithelium, is reprogrammed to NE-specific regulatory elements in NEPC. Despite loss of dependence upon AR, NEPC maintains FOXA1 expression and requires FOXA1 for proliferation and expression of NE lineage-defining genes. Ectopic expression of the NE lineage TFs ASCL1 and NKX2-1 in PRAD cells reprograms FOXA1 to bind to NE regulatory elements and induces enhancer activity as evidenced by histone modifications at these sites. Our data establish the importance of FOXA1 in NEPC and provide a principled approach to identifying cancer dependencies through epigenomic profiling.

New approach for direct analysis of opaque solutions with high content of suspended solids: Determination of calcium and phosphate in raw sugarcane juice.

This paper presents a new analytical approach for element concentration determination in samples containing significant concentrations of dissolved and suspended interferences. The proposed system enables to segregate of the complex matrix, species of interest from other interferences with a minimum requirement of reagents and energy. For this purpose, a new cleanup chamber design was implemented with cationic and anionic resins employed under membrane form and the tangential flow of the solution avoided the drawbacks commonly attributed to the packed and fluidized bed columns, such as the formation of preferred paths, increasing hydrodynamic pressure and clogging. The element concentration determination was colorimetrically performed with an automatic flow analysis system. The strategy was validated with the concentration determination of calcium and phosphorus in raw sugarcane juice. Quantification limit of 0.48 to calcium and 1.13 mg L-1 to phosphorus, linear range between 1 and 50 mg L-1, with RSD of 0.50 and 1.50% (n = 11) respectively.

Ethanol determination in fermented sugarcane substrates by a diffusive micro-distillation device.

The determination of ethanol in fermented substrates is an important parameter for monitoring the production of distilled beverage samples. The correct measurement of its content has a direct impact on the profitability of the process. In this work, a diffusive micro-distillation device (DMDD) is proposed that allows the determination of ethanol directly in the fermented or distilled beverages samples. The DMDD consists of a 5 mL plastic test tube containing a reagent solution of potassium dichromate and sulfuric acid, inserted into another 50 mL polyethylene tube containing the sample. This set is heated in a water bath for 15 min at 80 °C, providing the ethanol diffusion, which reacts with the receptor solution contained in the test tube. The chromium (III) produced by the oxidation reaction, is spectrophotometrically quantified at 589 nm. The proposed procedure has a linear range between 1 and 12% (v/v) with R2 = 0.999 and RSD = 3.8% and results in agreement with those obtained by the distillation-densitometry official method.

Optimization of ultrasound-assisted extraction of bioactive compounds from acerola waste.

The industrial processing of acerola (Malpighia emarginata D.C.) produces huge quantities of waste material that are badly discarded or undervalued. In spite of this, acerola wastes have a high content of antioxidant compounds. The aim of this work was to study the extraction of antioxidant compounds from acerola residues using ultrasound assisted extraction. Using multiple regression techniques, the effects of ethanol concentration in the hydroethanolic solution (C), extraction time (t), temperature (T), and liquid-solid ratio (R) on the total phenolic content, total flavonoid content and antioxidant potential were investigated. The best extraction conditions were identified using the desirability function, which is a multi-response optimization technique. The optimal processing parameters were 67.5% of ethanol concentration, temperature of 80.9 °C, liquid/solid ratio of 59.8 mL/g, and extraction time of 13.6 min. HPLC-UV has been used to identify the main antioxidant compounds obtained under these optimal condition. Based on the results, acerola waste has high potential for better use, such as in food and pharmaceutical applications.

A versatile X-ray phase retarder for lock-in XMCD measurements.

X-ray magnetic circular dichroism (XMCD) is a technique commonly used to probe magnetic properties of materials with element and orbital selectivity, which requires the use of circularly polarized (CP) X-rays. It is possible to accomplish XMCD experiments with fixed CP and alternating the magnetic field orientation, but most reliable data are obtained when alternating the magnetization orientation and the polarization between right and left helicities. A versatile strategy has been developed to perform XMCD experiments using a hard X-ray quarter-wave plate, at both polychromatic dispersive and conventional monochromatic optics, in combination with synchronous data acquisition. The switching frequency waveform is fed into a lock-in amplifier to detect and amplify the XMCD signal. The results on a reference sample demonstrate an improvement in data quality and acquisition time. The instrumentation successfully generated 98% of CP X-rays switching the beam helicity at 13 Hz, with the possibility of faster helicity switching once it is installed at the new Brazilian fourth-generation source, SIRIUS.

Impact of Drug-Polymer Interaction in Amorphous Solid Dispersion Aiming for the Supersaturation of Poorly Soluble Drug in Biorelevant Medium.

The aim of this study was to investigate the influence of the production method and the polymeric carrier on the ability to generate and maintain the supersaturation of a poorly soluble drug in biorelevant medium. The amorphous solid dispersion of sulfamethoxazole, an antibacterial drug, was produced using two different polymers by spray-drying or hot melt extrusion methods. When Eudragit EPO was used, supersaturation was maintained up to 24 h for both techniques at all drug-polymer proportions. However, when Soluplus was employed in hot melt extrusion, a smaller amount of drug was dissolved when compared to the amorphous drug. The proportion of 3:7 drug-Eudragit EPO (w/w) produced by spray-drying presented a higher amount of drug dissolved in supersaturation studies and it was able to maintain the physical stability under different storage conditions throughout the 90-day evaluation. Supersaturation generation and system stability were found to be related to more effective chemical interaction between the polymer and the drug provided by the production method, as revealed by the 1D ROESY NMR experiment. Investigation of drug-polymer interaction is critical in supersaturating drug delivery systems to avoid crystallization of the drug and to predict the effectiveness of the system. Chemical compounds studied in this article: Sulfamethoxazole (PubChem CID: 4539) and Methacrylate copolymer - Eudragit EPO (PubChem CID: 65358).

Prostate cancer reactivates developmental epigenomic programs during metastatic progression.

Epigenetic processes govern prostate cancer (PCa) biology, as evidenced by the dependency of PCa cells on the androgen receptor (AR), a prostate master transcription factor. We generated 268 epigenomic datasets spanning two state transitions-from normal prostate epithelium to localized PCa to metastases-in specimens derived from human tissue. We discovered that reprogrammed AR sites in metastatic PCa are not created de novo; rather, they are prepopulated by the transcription factors FOXA1 and HOXB13 in normal prostate epithelium. Reprogrammed regulatory elements commissioned in metastatic disease hijack latent developmental programs, accessing sites that are implicated in prostate organogenesis. Analysis of reactivated regulatory elements enabled the identification and functional validation of previously unknown metastasis-specific enhancers at HOXB13, FOXA1 and NKX3-1. Finally, we observed that prostate lineage-specific regulatory elements were strongly associated with PCa risk heritability and somatic mutation density. Examining prostate biology through an epigenomic lens is fundamental for understanding the mechanisms underlying tumor progression.

Non-coding somatic mutations converge on the PAX8 pathway in ovarian cancer.

The functional consequences of somatic non-coding mutations in ovarian cancer (OC) are unknown. To identify regulatory elements (RE) and genes perturbed by acquired non-coding variants, here we establish epigenomic and transcriptomic landscapes of primary OCs using H3K27ac ChIP-seq and RNA-seq, and then integrate these with whole genome sequencing data from 232 OCs. We identify 25 frequently mutated regulatory elements, including an enhancer at 6p22.1 which associates with differential expression of ZSCAN16 (P = 6.6 × 10-4) and ZSCAN12 (P = 0.02). CRISPR/Cas9 knockout of this enhancer induces downregulation of both genes. Globally, there is an enrichment of single nucleotide variants in active binding sites for TEAD4 (P = 6 × 10-11) and its binding partner PAX8 (P = 2×10-10), a known lineage-specific transcription factor in OC. In addition, the collection of cis REs associated with PAX8 comprise the most frequently mutated set of enhancers in OC (P = 0.003). These data indicate that non-coding somatic mutations disrupt the PAX8 transcriptional network during OC development.

Analysis of the use of a non-conventional rotary drum for dehydration of microalga Spirulina platensis.

The high content of bioactive compounds in the microalga Spirulina platensis has recently attracted attention from food and pharmaceutical industries. However, for its application an effective preservation technique must be developed. In this paper, we investigated the use of a non-conventional rotary dryer (with an inert bed) for drying the microalga Spirulina biomass and the effects of the operational conditions (air temperature, intermittent feeding interval, filling degree of inert particles, and rotation speed) on its bioactive compounds. The results indicated that this non-conventional drying system offers an effective alternative for expanding the use of this biomass in an adequate form. We identified the conditions in which the dried material had maintained satisfactory contents of phenolics (air temperature of 70 °C and intermittent feeding interval of 10 min), flavonoids (intermittent feeding interval of 17.4 min), and phycocyanin compounds (air temperature of 40 °C), which were near to those present in fresh microalga.