A population-based study of children suggests blunted morning cortisol rhythms are associated with alterations of the systemic inflammatory state

Background: In children, digital media, lifestyle, and the COVID pandemic have impacted sunlight exposure, exercise, and diet patterns - cues that entrain the circadian clock. We hypothesized that low morning cortisol reflects a weak circadian clock, impacting the pro-inflammatory state. The primary objective was to test relationships between diurnal cortisol fluctuations and the inflammatory state in children as a means of providing indirect support for this hypothesis. Methods: The Cardiovascular Health Intervention Program (CHIP) was a population-based cross-sectional and longitudinal study of circadian health in public elementary school children in Southern Maine, USA (recruitment period 2012–2017). Participants were 689 students in 4th grade (baseline; age=9.2 ± 0.4 years), and 647 students in 5th grade (age=10.5 ± 0.5 years). Nine salivary cortisol measures per child (2 awakening and 1 prior to bed for 3 sequential days) (n = 1336 child phenotype days; n = 7987 cortisol assays), 10 cytokines measured in morning and evening saliva samples (n = 202 child phenotype days), and lipids were measured. Clinical outcomes were blood pressure, weight and height (body mass index [BMI]; BMI = kg/m2), among others. Findings: Upon-waking cortisol levels were 0.28 ± 0.13 µg/dL, 30-minute post-waking 0.33 ± 0.15 µg/dL, and evening 0.08 ± 0.10 µg/dL. Salivary cytokine levels (n = 202) showed interleukins (IL) IL-1β and IL-8 were highest in early morning (upon awakening; AM), and IL-6 and tumor necrosis factor (TNF) TNF-α highest before bed (PM) (IL-1β AM \u3e PM [−4.02 fold; p \u3c 0.001]; IL-8 AM \u3e PM [−1.36 fold; p \u3c 0.001]; IL-6 AM \u3c PM [+1.49 fold; p \u3c 0.001]; TNF-α AM \u3c PM [+1.73 fold; p = 0.03]. Regression modeling showed high morning cortisol was associated with high morning IL-1β (p = 3.82 ×10−6), but low evening IL-1β (p = 6.27 ×10−4). Regression modeling of BMI z-score as the response variable showed the expected significant relationships to high density lipoprotein (HDL) (negative; p \u3c 0.001), mean arterial pressure (positive; p \u3c 0.001), and morning cortisol (negative; p = 0.01) but only weak relationships to either evening cortisol (p = 0.1) or cytokine (positive; p = 0.02; from the model with smallest Rsquared) levels. Interpretation: We provide preliminary data on diurnal fluctuations of inflammatory cytokines in saliva in a population-based cohort of children. Correlation of morning and evening cortisol levels with inflammatory cytokines in the same saliva samples showed that high morning cortisol was associated with high morning IL-1β and low evening IL-1β. Future studies may test the hypothesis that strong diurnal cycling of IL-1β may serve as a homeostatic mechanism keeping the immune system in check, and that low morning cortisol (possible circadian misalignment) may lead to less stringent control of inflammatory networks

A set of proteomic tools for the characterization of proteins from diverse organisms : plants, fungi and parasites

L’analyse protéomique par spectrométrie de masse s’est imposée comme une méthode incontournable pour la caractérisation des protéines. Grâce aux progrès de l’instrumentation et de la bioinformatique, l’interprétation automatisée des spectres MS/MS permet aujourd’hui d’identifier des milliers de protéines dans un type cellulaire. Cependant, cette méthodologie s’applique encore difficilement aux organismes dont les génomes n’ont pas été séquencés, et donc pour lesquels il n’existe pas de banques de séquences peptidiques de référence. Notre travail a porté sur le développement et l’application d’une méthodologie d’interprétation des données MS/MS pour les organismes à génomes non séquencés. Cette méthodologie est basée sur le séquençage de novo suivi de recherche MS-BLAST. Ainsi nous avons pu : Identifier les différents partenaires de complexes protéiques tels que les protéines des complexes TgGAP50, TgAlba, TgSORTLR impliqués dans la motilité, la virulence ou le trafic intracellulaire des protéines du parasite Toxoplasma gondii, Identifier et caractériser des variants d’hémoglobine humaine, Identifier les protéines différentiellement exprimées lors des interactions vigne et champignons à génomes non séquencés dans la maladie de l’esca, Caractériser finement la N-glycosylation de l’invertase vacuolaire du raisin. Nous avons pu réaliser nos études sur des échantillons d’origines très différentes : homme, plantes, champignons, parasites et nous avons apporté des éléments de réponses moléculaires aux questions biologiques.The proteomic analysis by mass spectrometry is now an essential method for the characterization of proteins. Thanks to advances in instrumentation and bioinformatics, automated interpretation of MS/MS spectra can now identify thousands of proteins in a cell type. However, this methodology remains poorly applied to the organisms that genomes are not sequenced and therefore where there is no database of reference for peptides sequences. Our work has focused on the development and application of a methodology for the interpretation of MS/MS data for the organisms that genomes are not sequenced. This methodology is based on the de novo sequencing followed by MS-BLAST search. Thus we have: Identify different partners of protein complexes such as proteins TgGAP50, TgAlba and TgSORTLR complex, involved in motility, virulence or intracellular protein trafficking of Toxoplasma gondii, Identify and characterize human hemoglobin variants, Identify the proteins differentially expressed during interaction of vines and fungi that genomes are not sequenced in esca disease, Finely characterize the N-glycosylation of the grape vacuolar invertase. We have achieved our studies on samples of very different origins: human, plants, fungi, parasites, and we provided evidence of molecular responses to biological questions

Un ensemble d'outils protéomiques pour la caractérisation de protéines d'organismes très divers : plantes, champignons et parasites

The proteomic analysis by mass spectrometry is now an essential method for the characterization of proteins. Thanks to advances in instrumentation and bioinformatics, automated interpretation of MS/MS spectra can now identify thousands of proteins in a cell type. However, this methodology remains poorly applied to the organisms that genomes are not sequenced and therefore where there is no database of reference for peptides sequences. Our work has focused on the development and application of a methodology for the interpretation of MS/MS data for the organisms that genomes are not sequenced. This methodology is based on the de novo sequencing followed by MS-BLAST search. Thus we have: Identify different partners of protein complexes such as proteins TgGAP50, TgAlba and TgSORTLR complex, involved in motility, virulence or intracellular protein trafficking of Toxoplasma gondii, Identify and characterize human hemoglobin variants, Identify the proteins differentially expressed during interaction of vines and fungi that genomes are not sequenced in esca disease, Finely characterize the N-glycosylation of the grape vacuolar invertase. We have achieved our studies on samples of very different origins: human, plants, fungi, parasites, and we provided evidence of molecular responses to biological questions.L’analyse protéomique par spectrométrie de masse s’est imposée comme une méthode incontournable pour la caractérisation des protéines. Grâce aux progrès de l’instrumentation et de la bioinformatique, l’interprétation automatisée des spectres MS/MS permet aujourd’hui d’identifier des milliers de protéines dans un type cellulaire. Cependant, cette méthodologie s’applique encore difficilement aux organismes dont les génomes n’ont pas été séquencés, et donc pour lesquels il n’existe pas de banques de séquences peptidiques de référence. Notre travail a porté sur le développement et l’application d’une méthodologie d’interprétation des données MS/MS pour les organismes à génomes non séquencés. Cette méthodologie est basée sur le séquençage de novo suivi de recherche MS-BLAST. Ainsi nous avons pu : Identifier les différents partenaires de complexes protéiques tels que les protéines des complexes TgGAP50, TgAlba, TgSORTLR impliqués dans la motilité, la virulence ou le trafic intracellulaire des protéines du parasite Toxoplasma gondii, Identifier et caractériser des variants d’hémoglobine humaine, Identifier les protéines différentiellement exprimées lors des interactions vigne et champignons à génomes non séquencés dans la maladie de l’esca, Caractériser finement la N-glycosylation de l’invertase vacuolaire du raisin. Nous avons pu réaliser nos études sur des échantillons d’origines très différentes : homme, plantes, champignons, parasites et nous avons apporté des éléments de réponses moléculaires aux questions biologiques

Un ensemble d'outils protéomiques pour la caractérisation de protéines d'organismes très divers (plantes, champignons et parasites)

L analyse protéomique par spectrométrie de masse s est imposée comme une méthode incontournable pour la caractérisation des protéines. Grâce aux progrès de l instrumentation et de la bioinformatique, l interprétation automatisée des spectres MS/MS permet aujourd hui d identifier des milliers de protéines dans un type cellulaire. Cependant, cette méthodologie s applique encore difficilement aux organismes dont les génomes n ont pas été séquencés, et donc pour lesquels il n existe pas de banques de séquences peptidiques de référence. Notre travail a porté sur le développement et l application d une méthodologie d interprétation des données MS/MS pour les organismes à génomes non séquencés. Cette méthodologie est basée sur le séquençage de novo suivi de recherche MS-BLAST. Ainsi nous avons pu : Identifier les différents partenaires de complexes protéiques tels que les protéines des complexes TgGAP50, TgAlba, TgSORTLR impliqués dans la motilité, la virulence ou le trafic intracellulaire des protéines du parasite Toxoplasma gondii, Identifier et caractériser des variants d hémoglobine humaine, Identifier les protéines différentiellement exprimées lors des interactions vigne et champignons à génomes non séquencés dans la maladie de l esca, Caractériser finement la N-glycosylation de l invertase vacuolaire du raisin. Nous avons pu réaliser nos études sur des échantillons d origines très différentes : homme, plantes, champignons, parasites et nous avons apporté des éléments de réponses moléculaires aux questions biologiques.The proteomic analysis by mass spectrometry is now an essential method for the characterization of proteins. Thanks to advances in instrumentation and bioinformatics, automated interpretation of MS/MS spectra can now identify thousands of proteins in a cell type. However, this methodology remains poorly applied to the organisms that genomes are not sequenced and therefore where there is no database of reference for peptides sequences. Our work has focused on the development and application of a methodology for the interpretation of MS/MS data for the organisms that genomes are not sequenced. This methodology is based on the de novo sequencing followed by MS-BLAST search. Thus we have: Identify different partners of protein complexes such as proteins TgGAP50, TgAlba and TgSORTLR complex, involved in motility, virulence or intracellular protein trafficking of Toxoplasma gondii, Identify and characterize human hemoglobin variants, Identify the proteins differentially expressed during interaction of vines and fungi that genomes are not sequenced in esca disease, Finely characterize the N-glycosylation of the grape vacuolar invertase. We have achieved our studies on samples of very different origins: human, plants, fungi, parasites, and we provided evidence of molecular responses to biological questions.STRASBOURG-Bib.electronique 063 (674829902) / SudocSudocFranceF

The Toxoplasma gondii dense granule protein TgGRA3 interacts with host Golgi and dysregulates anterograde transport

After entry into the host cell, the intracellular parasite Toxoplasma gondii resides within a membrane-bound compartment, the parasitophorous vacuole (PV). The PV defines an intracellular, parasite-specific niche surrounded by host organelles, including the Golgi apparatus. The mechanism by which T. gondii hijacks the host Golgi and subverts its functions remains unknown. Here, we present evidence that the dense granule protein TgGRA3 interacts with host Golgi, leading to the formation of tubules and the entry of host Golgi material into the PV. Targeted disruption of the TgGRA3 gene delays this engulfment of host Golgi. We also demonstrate that TgGRA3 oligomerizes and binds directly to host Golgi membranes. In addition, we show that TgGRA3 dysregulates anterograde transport in the host cell, thereby revealing one of the mechanisms employed by T. gondii to recruit host organelles and divert their functions. This article has an associated First Person interview with the first author of the paper

In-depth glycoproteomic characterisation of grape berry vacuolar invertase using a combination of mass spectrometry-based approaches

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Proteomic and Bioinformatic Analysis of Decellularized Pancreatic Extracellular Matrices

Tissue microenvironments are rich in signaling molecules. However, factors in the tissue matrix that can serve as tissue-specific cues for engineering pancreatic tissues have not been thoroughly identified. In this study, we performed a comprehensive proteomic analysis of porcine decellularized pancreatic extracellular matrix (dpECM). By profiling dpECM collected from subjects of different ages and genders, we showed that the detergent-free decellularization method developed in this study permits the preservation of approximately 62.4% more proteins than a detergent-based method. In addition, we demonstrated that dpECM prepared from young pigs contained approximately 68.5% more extracellular matrix proteins than those prepared from adult pigs. Furthermore, we categorized dpECM proteins by biological process, molecular function, and cellular component through gene ontology analysis. Our study results also suggested that the protein composition of dpECM is significantly different between male and female animals while a KEGG enrichment pathway analysis revealed that dpECM protein profiling varies significantly depending on age. This study provides the proteome of pancreatic decellularized ECM in different animal ages and genders, which will help identify the bioactive molecules that are pivotal in creating tissue-specific cues for engineering tissues in vitro

Physiological Changes in Green Stems of Vitis vinifera L. cv. Chardonnay in Response to Esca Proper and Apoplexy Revealed by Proteomic and Transcriptomic Analyses

International audienc

Proteomic insights into changes in grapevine wood in response to esca proper and apoplexy

Among fungal grapevine trunk diseases, esca proper poses a significant threat for viticulture. Apoplexy, mainly occurring on grapevines affected by esca proper, is also a threat. To verify if different responses are activated in the woody tissues of apoplectic (A) and esca proper-affected (E) vines, two-dimensional gel electrophoresis coupled to mass spectrometry analysis was used to examine changes in the trunk wood of E and A field-grown plants. Asymptomatic and black streaked trunk (BST) wood from A and E plants were compared to asymptomatic and BST wood of control plants. Twenty-seven differentially expressed protein spots were identified. For eleven targeted proteins, expression of the relative transcripts was also monitored by qRT-PCR. Hierarchical tree clustering revealed differences in the distribution of spots containing carbohydrate metabolism proteins and heat shock proteins between asymptomatic- and BST-wood of grapevine, irrespective of the type of plant examined (control or diseased grapevines). Asymptomatic wood was mainly characterized by down-expression of proteins involved in cell growth and defense responses. The proteome of BST wood, characterized by extensive presence of grapevine trunk disease agents, revealed over-expression of proteins involved in defense. There was no evidence of strong different response in the trunk wood of apoplectic and esca proper affected plants. This could mean that, despite the different feature of these external crown symptoms, grapevine responses at trunk wood level is very similar in both cases. This plant response might therefore either simply be due to the fact that plants can react in the same way to different stresses, or that apoplexy represents a different effect provoked by the same cause or causal agent(s)