Prolonged sleep deprivation induces a cytokine-storm-like syndrome in mammals.

Most animals require sleep, and sleep loss induces serious pathophysiological consequences, including death. Previous experimental approaches for investigating sleep impacts in mice have been unable to persistently deprive animals of both rapid eye movement sleep (REMS) and non-rapid eye movement sleep (NREMS). Here, we report a "curling prevention by water" paradigm wherein mice remain awake 96% of the time. After 4 days of exposure, mice exhibit severe inflammation, and approximately 80% die. Sleep deprivation increases levels of prostaglandin D2 (PGD2) in the brain, and we found that elevated PGD2 efflux across the blood-brain-barrier-mediated by ATP-binding cassette subfamily C4 transporter-induces both accumulation of circulating neutrophils and a cytokine-storm-like syndrome. Experimental disruption of the PGD2/DP1 axis dramatically reduced sleep-deprivation-induced inflammation. Thus, our study reveals that sleep-related changes in PGD2 in the central nervous system drive profound pathological consequences in the peripheral immune system.

Widely targeted metabolomics reveal the glucosinolate profile and odor-active compounds in flowering Chinese cabbage powder.

Widely targeted metabolomics were performed to explore the differences in glucosinolate and odor-active compound levels between flowering Chinese cabbage powder (FCCP) under vacuum-drying and oven-drying conditions. Twenty-three aliphatic, five indole, and three aromatic glucosinolates were identified in two pretreated FCCP. Higher aliphatic glucosinolate levels were retained in vacuum-dried cabbage powder compared to oven-dried samples, and they were negatively correlated with treated temperatures. A total of 36 major odor contributing compounds were detected, including 5 sulfur compounds, 10 aldehydes, 9 heterocyclic compounds, 7 nitriles, 3 acids, and 2 others. 5-Hexenenitrile and (methyldisulfanyl)methan, provide typical pungent, sulfous, and vegetable notes in FCCP. Four major GSLs, namely 2(R)-hydroxy-3-butenyl glucosinolate, (2S)-2-hydroxy-4-pentenyl glucosinolate, 5-(methylthio)pentyl glucosinolate and 2-phenylethyl glucosinolate were the key precursors to form odor-active compounds. Higher temperatures in thermal effects promotes the formation of sulfur-containing and nitrile compounds compared to the vacuum-dried ones. This work can provide a guide for flavor and nutrition retention in FCCP process.

Flavor characteristics of peeled walnut kernels under two-steps roasting processes.

Currently, the effects of roasting methods on the flavor profile of peeled walnut kernels (PWKs) remain unknown. The effects of hot air binding (HAHA), radio frequency (HARF), and microwave irradiation (HAMW) on PWK were evaluated using olfactory, sensory, and textural techniques. Solvent Assisted Flavor Evaporation-Gas Chromatography-Olfactometry (SAFE-GC-O) identified 21 odor-active compounds with total concentrations of 229 µg/kg, 273 µg/kg and 499 µg/kg due to HAHA, HARF, and HAMW, respectively. HAMW exhibited the most prominent nutty taste, with the highest response among roasted milky sensors with the typical aroma of 2-ethyl-5-methylpyrazine. HARF had the highest values for chewiness (5.83 N·mm) and brittleness (0.68 mm); however, these attributes did not contribute to the flavor profile. The partial least squares regression (PLSR) model and VIP values showed 13 odor-active compounds were responsible for the sensory differences from different processes. The two-step treatment with HAMW improved the flavor quality of PWK.

A high-performance genetically encoded fluorescent indicator for in vivo cAMP imaging.

cAMP is a key second messenger that regulates diverse cellular functions including neural plasticity. However, the spatiotemporal dynamics of intracellular cAMP in intact organisms are largely unknown due to low sensitivity and/or brightness of current genetically encoded fluorescent cAMP indicators. Here, we report the development of the new circularly permuted GFP (cpGFP)-based cAMP indicator G-Flamp1, which exhibits a large fluorescence increase (a maximum ΔF/F0 of 1100% in HEK293T cells), decent brightness, appropriate affinity (a Kd of 2.17 µM) and fast response kinetics (an association and dissociation half-time of 0.20 and 0.087 s, respectively). Furthermore, the crystal structure of the cAMP-bound G-Flamp1 reveals one linker connecting the cAMP-binding domain to cpGFP adopts a distorted ß-strand conformation that may serve as a fluorescence modulation switch. We demonstrate that G-Flamp1 enables sensitive monitoring of endogenous cAMP signals in brain regions that are implicated in learning and motor control in living organisms such as fruit flies and mice.

Flavor characteristics of peanut butter pretreated by radio frequency heating, explosion puffing, microwave, and oven heating.

Currently, the effect of different pretreatments (i.e., radio frequency (RF), explosion puffing (EP), microwave (MW) and oven heating (OH)) on the flavor characteristics of peanut butter is unclear. Consequently, this study identified volatile aroma and non-volatile taste using HS-SPME/GC-MS combined with the use of an electronic nose, electronic tongue, and sniffing. 53 volatile compounds in four peanut butters were identified, MW-treated samples exhibited the most aroma-active compounds (43), followed by samples treated using OH (42), EP (38) and RF (21). Different pretreatment resulted in significant flavor differences in the aroma and taste. The peanut butter under MW pretreatment had a strongest nutty notes among the treatments. RF methods yielded smaller particle sizes and better texture compared to conventional OH. However, instantaneous heating using EP did not result in improvements to the aroma or taste. A combination of MW and RF may improve the flavor quality of peanut butter.

Analysis of equivalent thickness of geological media for lab-scale study of radon exhalation.

Geological media are omnipresent in nature. Lab-scale tests are frequently employed in radon exhalation measurements for these media. Thus, it is critical to find the thickness of the medium at an experimental scale that is equivalent to the medium thickness in a real geological system. Based on the diffusion-advection transport of radon, theoretical models of the surface radon exhalation rate for homogeneous semi-infinite and finite-thickness systems were derived (denoted as Jse and Jfi, respectively). Analysis of the equivalency of Jse and Jfi was subsequently carried out by introducing several dimensionless parameters, including the ratio of the exhalation rates for the semi-infinite and finite-thickness models, ε, and the number of diffusion lengths required to achieve a desired ε value, n. The results showed that when radon transport in geological media is dominantly driven by diffusion effect, if n > 3.6626, then ε > 95%; if n > 5.9790, then ε > 99.5%. When radon migration is dominantly driven by advection effect, if n > 2.5002, then ε > 95%; if n > 4.0152, then ε > 99.5%. Therefore, if the thickness of the geological media (x0) is greater than a certain n times the radon diffusion length of the media (L), the media can be modeled as semi-infinite. To validate the model, a pure radon diffusion experiment (no advection) was developed using uranium mill tailings, laterite, and radium-bearing rocklike material with different thicknesses (x0). The theoretical model was demonstrated to be reliable and valid. This study provides a basis for determining the appropriate thickness of geological media in lab-scale radon exhalation measurement experiments with open bottom.

Identification of volatile sulfur-containing compounds and the precursor of dimethyl sulfide in cold-pressed rapeseed oil by GC-SCD and UPLC-MS/MS.

Volatile sulfur-containing compounds (VSCs) provide an important contribution to foods due to their special odors. In this study, VSCs in 21 cold-pressed rapeseed oils (CROs) from 9 regions in China were extracted and separated by headspace solid-phase microextraction combined with gas chromatography coupled with sulfur chemiluminescence detection. 19 VSCs were identified by authentic standards, and the total concentration of VSCs in all CROs ranged from 49.0 to 18129 µg/kg. Dimethyl sulfide (DMS), with its high odor activity value (7-14574), was the most significant aroma contributor to the CROs. Furthermore, S-methylmethionine (SMM) in rapeseed was first affirmed by ultra-performance liquid chromatography-tandem mass spectrometry and isotope quantitation. The positive correlation coefficient between DMS and SMM was 0.793 (p < 0.05), which confirmed SMM as a crucial precursor of DMS in CROs. This study provided a theoretical basis for selecting rapeseed materials by the distribution of essential VSCs and the source of DMS.

Investigation of volatile thiol contributions to rapeseed oil by odor active value measurement and perceptual interactions.

Volatile thiols are important aroma components of rapeseed oil. This study established an identification and quantification method of volatile thiols via headspace solid-phase microextraction and gas chromatography-sulfur chemiluminescence detection. Four thiols (phenylmethanthiol, 3-sulfanyl-1-hexanol, 2-methyl-3-furanthiol, and 2-furylmethanthiol) were newly identified in microwaved rapeseed oil, and cause sesame, roasted meat, and garlic odors. The total concentration of the four thiols in rapeseed oil obtained from 13 rapeseed varieties ranged from 11.47 to 153.72 µg/kg. Determination of the threshold revealed that 3-sulfanyl-1-hexanol possessed the highest odor active value (7565), followed by phenylmethanthiol (3589), 2-furylmethanthiol (626), and 2-methyl-3-furanthiol (28). Further, perceptual interactions between volatile thiols and characteristic odor (3-butenyl isothiocyanate) of rapeseed oil were evaluated by Feller's addition model and S-curve method, which revealed that 2-methyl-3-furanthiol, 2-furylmethanthiol, phenylmethanthiol, and 3-sulfanyl-1-hexanol present a positive effect with 3-butenyl isothiocyanate. This study provides deep insights into the impact of sulfur-containing compounds on the aroma of rapeseed oil.

Species Diversity and Chemotypes of Fusarium Species Associated With Maize Stalk Rot in Yunnan Province of Southwest China.

Maize stalk rot caused by Fusarium species is one of the most important fungal diseases of maize throughout the world. The disease is responsible for considerable yield losses and has also been associated with mycotoxin contamination of the crop. In this study, a survey of maize stalk rot was performed in seven locations of Yunnan Province in China during the cropping season of 2015 and 2016. Based on morphological and molecular characteristics, 204 isolates belonging to 12 Fusarium spp. from symptomatic stalks of maize were identified. Among the isolated strains, 83 were identified as Fusarium meridionale (40.5%), 46 as Fusarium boothii (22.5%), 34 as Fusarium temperatum (16.5%), 12 as Fusarium equiseti (5.9%), 10 as Fusarium asiaticum (4.9%), six as Fusarium proliferatum (3.0%), four as Fusarium verticillioides (2.0%), four as Fusarium incarnatum (2.0%), two as Fusarium avenaceum (1.0%), one as Fusarium cerealis (0.5%), one as Fusarium graminearum (0.5%), and one as Fusarium cortaderiae (0.5%). Fusarium cortaderiae was the first report on the causal agent of maize stalk rot disease in China. These isolates were divided into five chemotypes: nivalenol (NIV), deoxynivalenol (DON), beauvericin (BEA), zearalenone (ZEN), and fumonisin (FUM). Phylogenetic analysis based on partial sequences of the translation elongation factor 1α (TEF1-α) showed a high degree of interspecific polymorphisms among the isolates. Pathogenicity analysis on maize stalks indicated that all the 12 species of Fusarium were able to cause the disease symptoms with different aggressiveness. This study on population, pathogenicity, and toxigenic chemotypes of Fusarium species associated with maize stalk rot in Yunnan Province of southwest China, will help design an effective integrated control strategy for this disease.

Hi-C profiling of cancer spheroids identifies 3D-growth-specific chromatin interactions in breast cancer endocrine resistance.

BACKGROUND: Organoids or spheroids have emerged as a physiologically relevant in vitro preclinical model to study patient-specific diseases. A recent study used spheroids of MCF10 cells to model breast cancer progression and identified targetable alterations more similar to those in vivo. Thus, it is practical and essential to explore and characterize the spheroids of the commonly used human breast cancer (BC) cells. METHODS: In this study, we conducted Hi-C analyses in three-dimensional (3D) spheroids of MCF10A, MCF7 and MCF7TR cells and compared TADs and looping genes with those in 2D monolayers. Furthermore, we performed in silico functional analysis on 3D-growth-specific looping genes and to compare patient outcomes with or without endocrinal therapy. Finally, we performed 3C/RT-qPCR validations in 3D spheroids and 3D-FISH confirmations in organoids of breast cancer patient tissues. RESULTS: We found that chromatin structures have experienced drastic changes during the 3D culture growth of BC cells although there is not much change in the quantity of chromatin domains. We also observed that the strengths of looping genes were statistically different between 2D monolayers and 3D spheroids. We further identified novel 3D growth-specific looping genes within Hippo relevant pathways, of which two genes showed potential prognostic values in measuring the outcome of the endocrine treatment. We finally confirmed a few selected genes in Hippo relevant pathways with enhanced looping in organoids of breast cancer patient tissues. CONCLUSIONS: Hence, our work has provided significant insights into our understanding of 3D-growth-specific chromatin architecture in tamoxifen-resistant breast cancer. Our analyses suggest that the strengthened looping-mediated Hippo relevant pathways may contribute to endocrine therapy resistance in breast cancer patients.

Unraveling of the Aroma-Active Compounds in Virgin Camellia Oil (Camellia oleifera Abel) Using Gas Chromatography-Mass Spectrometry-Olfactometry, Aroma Recombination, and Omission Studies.

Camellia oil is a popular edible oil in China as a result of its nutritional properties. However, the key odorants of camellia oil remain unclear. In this study, the volatiles of virgin camellia oil (VCO) were extracted by solvent-assisted and non-solvent-assisted methods. A total of 66 volatile compounds were identified using gas chromatography-mass spectrometry-olfactometry, with flavor dilution factors ranging from 1 to 729 via aroma extraction dilution analysis. Among them, 10 odorants were identified for the first time in VCO. Moreover, 41 volatiles were confirmed as aroma-active compounds with odor activity values greater than 1. Aroma recombination and omission studies demonstrated that aldehydes, esters, acids, and heterocyclic compounds significantly contribute to the aroma profiles of VCO. Hexanal, octanal, (E,E)-2,4-heptadienal, (E,E)-2,4-nonadienal, decyl acetate, ethyl benzoate, ethyl 2-methylbutanoate, 2-methylbutyl (Z)-2-methyl-2-butenoate, 2-methylbutanoic acid, hexanoic acid, 2-pentylfuran, and 2-methyl-3-furanthiol could impart roasted-like, nut-like, fat-like, fruit-like, grass-like, and sweat-like odors and were the key odorants in VCO. The lipoxygenase pathway was possibly responsible for the formation of key odorants in VCO. This work provides an extract aroma consistent for virgin camellia oil.

Key Odorant Differences in Fragrant Brassica napus and Brassica juncea Oils Revealed by Gas Chromatography-Olfactometry, Odor Activity Values, and Aroma Recombination.

Fragrant Brassica species seed oils (FBO) produced in China are mainly obtained from rapeseed (Brassica napus: B. napus) and mustard seeds (Brassica juncea: B. juncea). The characterization and differences of aroma profiles between those two species remain unclear. In this study, the volatile compounds in FBOs were systemically extracted by headspace solid-phase microextraction and solvent-assisted flavor evaporation combined with ultrasound and identified by comprehensive two-dimensional gas chromatography and time-of-flight mass spectrometry (GC×GC-TOFMS) and gas chromatography-olfactometry (GC-O). Ninety-three odorants were identified as aroma-active compounds with flavor dilution (FD) factors ranging from 1 to 6561. Moreover, 63 key compounds exhibited their odor activity values (OAVs) to be greater than 1. The oils of the two species were successfully recombinated with their key odorants. B. juncea oils presented stronger pungent-like, pickled-like, and fishy like notes compared to B. napus oils. The key odor differences were primarily attributed to the concentration of 3-butenenitrile, 4-(methylsulfanyl)butanenitrile, 5-(methylsulfanyl)pentanenitrile, 3-isothiocyanato-1-propene, 3-methyl-3-butenenitrile, isothiocyanatocyclopropane, (methylsulfanyl)acetonitrile, dimethyl sulfide, dimethyl trisulfide, and 3-(methyldisulfanyl)-1-propene. This work provides a guide for the selection of raw materials and odor markers in fragrant B. napus and B. juncea oils.

The 3D genomic landscape of differential response to EGFR/HER2 inhibition in endocrine-resistant breast cancer cells.

BACKGROUND: Recent studies suggested that crosstalk between ERα and EGFR/HER2 pathways plays a critical role in mediating endocrine therapy resistance. Several inhibitors targeting EGFR/HER2 signaling, including FDA-approved lapatinib and gefitinib as well as a novel dual tyrosine kinase inhibitor (TKI) sapitinib, showed greater therapeutic efficacies. However, how 3D chromatin landscape responds to the inhibition of EGFR/HER2 pathway remains to be elucidated. METHODS: In this study, we conducted in situ Hi-C and RNA-seq in two ERα+ breast cancer cell systems, 1) parental MCF7 cells and its associated tamoxifen-resistant MCF7TR cells; and 2) parental T47D cells and its associated tamoxifen-resistant T47DTR cells, before and after the treatment of sapitinib. RESULTS: We identified differential responses in topologically associated domains (TADs), looping genes and expressed genes. Interestingly, we found that many differential TADs and looping genes are reversible after sapitinib treatment, indicating that EGFR/HER2 signaling may play a role in reshaping and rewiring the high order genome organization. We further examined and recapitulated the reversible looping genes in 3D spheroids of breast cancer cells, demonstrating that 3D cell culture spheroid of breast cancer cells could be a potential preclinical breast cancer model for studying 3D chromatin regulation. CONCLUSIONS: Our study has provided significant insights into our understanding of 3D genomic landscape changes in response to EGFR/HER2 Inhibition in endocrine-resistant breast cancer cells. Our data provides a rich resource for further evaluating chromatin structural responses to EGFR/HER2 targeted therapies in endocrine-resistant breast cancer cells. Our analyses suggest that these alterations of chromatin structures and transcriptional programs may provide new avenues for intervention or designing of patient selection for targeted endocrine treatment.

Modeling and analysis of Hi-C data by HiSIF identifies characteristic promoter-distal loops.

Current computational methods on Hi-C analysis focused on identifying Mb-size domains often failed to unveil the underlying functional and mechanistic relationship of chromatin structure and gene regulation. We developed a novel computational method HiSIF to identify genome-wide interacting loci. We illustrated HiSIF outperformed other tools for identifying chromatin loops. We applied it to Hi-C data in breast cancer cells and identified 21 genes with gained loops showing worse relapse-free survival in endocrine-treated patients, suggesting the genes with enhanced loops can be used for prognostic signatures for measuring the outcome of the endocrine treatment. HiSIF is available at https://github.com/yufanzhouonline/HiSIF .

Author Correction: Temporal dynamic reorganization of 3D chromatin architecture in hormone-induced breast cancer and endocrine resistance.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Free and Bound Aroma Compounds of Turnjujube (Hovenia acerba Lindl.) during Low Temperature Storage.

Free and bound aroma volatiles from turnjujube during low temperature storage were extracted by headspace solid-phase microextraction. They were then characterized and identified using gas chromatography-mass spectrometry. Turnjujube was harvested and stored for 7, 14, and 21 days at 7 °C, the common temperature of display refrigerators in grocery stores. The results showed that 41 free and 24 bound aroma compounds were detected for the first time in turnjujube in both freshly harvested and stored turnjujube. The free and bound aroma compounds of turnjujube were markedly influenced by the storage time. The major free aroma compounds in turnjujube included esters, alcohols, aliphatic aldehydes, and aliphatic ketones. The major bound aroma compounds included borneol, eugenol, and isoeugenol, which contributed to sweet, floral, and herbaceous aroma after their hydrolysis. Freshly harvested turnjujube mostly had a fruity and herbaceous aroma, which diminished after storage at 7 °C. In contrast, the fatty aroma enhanced gradually over storage, and the floral aroma enhanced noticeably after storage for seven days. Foul odor was not detected even after storage at 7 °C for 21 days. The formation mechanisms of some aroma compounds were proposed.

Free and bound volatile compounds in the Rubus coreanus fruits of different ripening stages.

The aim of the present study was to investigate the free and bound volatiles in the Rubus coreanus (RC) fruits of different ripening stages. Thirty-seven free volatiles and 28 bound volatiles were identified in RC fruit for the first time. The contents of free (E)-2-hexen-1-ol, 1-hexanol, 2-heptanol, ß-myrcene, (E), (Z)-ß-ocimene, allo-ocimene, linalool, cosmene, α-terpineol, methyl salicylate, eugenol, and ß-damascenone remain high, and increased with the ripening of RC fruit. The contents of 11 bound volatiles decreased during the ripening, and became lower than the contents of their free volatiles in the ripe fruit. The ripe black fruit is closely correlated to the free nonanal, sulcatone, (E)-2-hexen-1-ol, 1-hexanol, 2-heptanol, 1-heptanol, 1-nonanol, (E)-linalool oxide (furanoid), and ß-damascenone, and bound (E)-2-hexen-1-ol and (E)- ß-ocimene. The ripe RC fruit is more fruity and floral than unripe fruit. The gradually hydrolyzed bound volatiles can enhance the fruity, floral, and herbaceous odors. PRACTICAL APPLICATIONS: Rubus coreanus (RC) fruit is a functional natural fruit. Both fresh and processed Rubus coreanus fruits including jams, confitures, wine, yogurt, vinegar, and beverages, as well as ingredients in functional foods or cosmetics have been extensively consumed. However, the free and bound aroma compounds in RC fruit have not been well understood. This work illustrates the contributions of free and bound volatiles to the flavor of RC fruit.

Temporal dynamic reorganization of 3D chromatin architecture in hormone-induced breast cancer and endocrine resistance.

Recent studies have demonstrated that chromatin architecture is linked to the progression of cancers. However, the roles of 3D structure and its dynamics in hormone-dependent breast cancer and endocrine resistance are largely unknown. Here we report the dynamics of 3D chromatin structure across a time course of estradiol (E2) stimulation in human estrogen receptor α (ERα)-positive breast cancer cells. We identified subsets of temporally highly dynamic compartments predominantly associated with active open chromatin and found that these highly dynamic compartments showed higher alteration in tamoxifen-resistant breast cancer cells. Remarkably, these compartments are characterized by active chromatin states, and enhanced ERα binding but decreased transcription factor CCCTC-binding factor (CTCF) binding. We finally identified a set of ERα-bound promoter-enhancer looping genes enclosed within altered domains that are enriched with cancer invasion, aggressiveness or metabolism signaling pathways. This large-scale analysis expands our understanding of high-order temporal chromatin reorganization underlying hormone-dependent breast cancer.

Changes of the free and bound volatile compounds in Rubus corchorifolius L. f. fruit during ripening.

The changes of free and bound volatile compounds in Rubus corchorifolius fruit during ripening were determined with a headspace SPME-GC-MS method. The results suggest that the free aldehydes, alcohols, esters and phenols increases, while that of free terpenoids decreases, with the ripening of the fruit. The bound aldehydes, alcohols, terpenoids, esters and phenols gradually decreases during ripening because these bound compounds are hydrolyzed to their free form. The characteristic free aroma compounds of ripened red fruit were found to be hexanal, 2-heptanone, ethyl hexanoate, 4-terpineol, geranial and methyleugenol. The free aroma compounds in red and yellow fruits exhibit similar odor profiles, and both of them are much sweeter, more floral and greener than the green fruit. The overall aroma of the fruits all ripening stages are mainly attributed to the free aroma compounds including ß-damascenone, hexanal, 2-hexenal and linalool. The formation mechanisms of some volatile compounds were proposed.

High strength gelatin-based nanocomposites reinforced by surface-deacetylated chitin nanofiber networks.

In this study, chitin nanofiber (ChNF) was deacetylated on the crystalline surface by NaOH treatment, leading to the fibrillation of mostly individualized nanofibers with high aspect ratio. The small diameter and high strength of chitin nanofibers make them promising reinforcing fillers for composites. Herein by introducing into the gelatin, surface-deacetylated chitin nanofiber (S-ChNF)/gelatin nanocomposites were fabricated in different component ratios using immersion method followed with drying. Due to the reinforcing effect attributed to S-ChNF, mechanical properties of the S-ChNF/gelatin were significantly improved in both stress and Young's modulus while still maintaining high transparency regardless of nanofiber content. Morphology and Fourier-transform infrared characterization revealed that S-ChNF preserved nanonetwork structures in the gelatin matrix and exhibited good compatibility through hydrogen bonding, which further confirmed the improvement in mechanical properties. Therefore, these S-ChNF/gelatin nanocomposites based on biocompatible and biodegradable raw materials have potential applications in biomedical and food packaging industries.