Validation of Messenger Ribonucleic Acid Markers Differentiating Among Human Acute Respiratory Distress Syndrome Subgroups in an Ovine Model of Acute Respiratory Distress Syndrome Phenotypes

BACKGROUND: The discovery of biological subphenotypes in acute respiratory distress syndrome (ARDS) might offer a new approach to ARDS in general and possibly targeted treatment, but little is known about the underlying biology yet. To validate our recently described ovine ARDS phenotypes model, we compared a subset of messenger ribonucleic acid (mRNA) markers in leukocytes as reported before to display differential expression between human ARDS subphenotypes to the expression in lung tissue in our ovine ARDS phenotypes model (phenotype 1 (Ph1): hypoinflammatory; phenotype 2 (Ph2): hyperinflammatory). METHODS: We studied 23 anesthetized sheep on mechanical ventilation with observation times between 6 and 24 h. They were randomly allocated to the two phenotypes (n = 14 to Ph1 and n = 9 to Ph2). At study end, lung tissue was harvested and preserved in RNAlater. After tissue homogenization in TRIzol, total RNA was extracted and custom capture and reporter probes designed by NanoString Technologies were used to measure the expression of 14 genes of interest and the 6 housekeeping genes on a nCounter SPRINT profiler. RESULTS: Among the 14 mRNA markers, in all animals over all time points, 13 markers showed the same trend in ovine Ph2/Ph1 as previously reported in the MARS cohort: matrix metalloproteinase 8, olfactomedin 4, resistin, G protein-coupled receptor 84, lipocalin 2, ankyrin repeat domain 22, CD177 molecule, and transcobalamin 1 expression was higher in Ph2 and membrane metalloendopeptidase, adhesion G protein-coupled receptor E3, transforming growth factor beta induced, histidine ammonia-lyase, and sulfatase 2 expression was higher in Ph1. These expression patterns could be found when different sources of mRNA – such as blood leukocytes and lung tissue – were compared. CONCLUSION: In human and ovine ARDS subgroups, similar activated pathways might be involved (e.g., oxidative phosphorylation, NF-κB pathway) that result in specific phenotypes

Low flow rate alters haemostatic parameters in an ex-vivo extracorporeal membrane oxygenation circuit

Extracorporeal membrane oxygenation (ECMO) is a life-saving modality used to manage cardiopulmonary failure refractory to conventional medical and surgical therapies. Despite advances in ECMO equipment, bleeding and thrombosis remain significant complications. While the flow rate for ECMO support is well recognized, less is known about the minimum-rate requirements and haemostasis. We investigated the relationship between different ECMO flow rates, and their effect on haemolysis and coagulation.Ten ex-vivo ECMO circuits were tested using donated

Reciprocal epithelial: Endothelial paracrine interactions during thyroid development govern follicular organization and C-cells differentiation.

The thyroid is a highly vascularized endocrine gland, displaying a characteristic epithelial organization in closed spheres, called follicles. Here we investigate how endothelial cells are recruited into the developing thyroid and if they control glandular organization as well as thyrocytes and C-cells differentiation. We show that endothelial cells closely surround, and then invade the expanding thyroid epithelial cell mass to become closely associated with nascent polarized follicles. This close and sustained endothelial:epithelial interaction depends on epithelial production of the angiogenic factor, Vascular Endothelial Growth Factor-A (VEGF-A), as its thyroid-specific genetic inactivation reduced the endothelial cell pool of the thyroid by >90%. Vegfa KO also displayed decreased C-cells differentiation and impaired organization of the epithelial cell mass into follicles. We developed an ex vivo model of thyroid explants that faithfully mimicks bilobation of the thyroid anlagen, endothelial and C-cells invasion, folliculogenesis and differentiation. Treatment of thyroid explants at e12.5 with a VEGFR2 inhibitor ablated the endothelial pool and reproduced ex vivo folliculogenesis defects observed in conditional Vegfa KO. In the absence of any blood supply, rescue by embryonic endothelial progenitor cells restored folliculogenesis, accelerated lumen expansion and stimulated calcitonin expression by C-cells. In conclusion, our data demonstrate that, in developing mouse thyroid, epithelial production of VEGF-A is necessary for endothelial cells recruitment and expansion. In turn, endothelial cells control epithelial reorganization in follicles and C-cells differentiation

Chamigny (Seine-et-Marne), Rue de la Marne - RD 80 : lieu-dit "La Grande Maison" : rapport de diagnostic

Un projet d'aménagement de lotissement déposé par la société Nexity sur la commune de Chamigny a conduit à la prescription d'un diagnostic archéologique par le Service régional d'Archéologie (DRAC, Île-de-France).Portant sur une surface totale prescrite de 22 044 m2, ce diagnostic a été réalisé par l'Inrap de mi-septembre à début octobre 2015.10,79 % de l'emprise prescrite ont été sondés sous forme de tranchées, de sondages profonds et d'ouvertures complémentaires. Ce diagnostic a permis la mise au jour de quatre sites chronologiques distincts. En premier, lieu, un niveau paléolithique moyen a été découvert lors de l'ouverture des sondages profonds 2 et 3, à une profondeur de 2,75 m et sur une épaisseur de 0,25 m. Ensuite, la présence d'une fosse datée du Néolithique ancien et plus particulièrement de la culture Blicquy-Villeneuve-Saint-Germain indique la présence d'une occupation de type habitat. Une sépulture collective de 5 m de long sur 3 m de large, dont l'état de conservation parait médiocre, dont est au moins attestée à deux étapes différentes du Néolithique final (IIIe millénaire avant J.-C.) avec une réutilisation à l'Epi-Campaniforme, a été mise au jour.Enfin, une très forte présence du haut Moyen Âge, dont la période mérovingienne est la plus représentée, a été attestée sur toute l'emprise. Considérant les découvertes faites pour cette période (notamment un four de potier et des indices concernant la métallurgie des non-ferreux), il se peut qu'il s'agisse d'un secteur artisanal au sein d'un habitat

Chamigny (Seine-et-Marne), Rue de la Marne - RD 80 : lieu-dit "La Grande Maison" : rapport de diagnostic

Un projet d'aménagement de lotissement déposé par la société Nexity sur la commune de Chamigny a conduit à la prescription d'un diagnostic archéologique par le Service régional d'Archéologie (DRAC, Île-de-France).Portant sur une surface totale prescrite de 22 044 m2, ce diagnostic a été réalisé par l'Inrap de mi-septembre à début octobre 2015.10,79 % de l'emprise prescrite ont été sondés sous forme de tranchées, de sondages profonds et d'ouvertures complémentaires. Ce diagnostic a permis la mise au jour de quatre sites chronologiques distincts. En premier, lieu, un niveau paléolithique moyen a été découvert lors de l'ouverture des sondages profonds 2 et 3, à une profondeur de 2,75 m et sur une épaisseur de 0,25 m. Ensuite, la présence d'une fosse datée du Néolithique ancien et plus particulièrement de la culture Blicquy-Villeneuve-Saint-Germain indique la présence d'une occupation de type habitat. Une sépulture collective de 5 m de long sur 3 m de large, dont l'état de conservation parait médiocre, dont est au moins attestée à deux étapes différentes du Néolithique final (IIIe millénaire avant J.-C.) avec une réutilisation à l'Epi-Campaniforme, a été mise au jour.Enfin, une très forte présence du haut Moyen Âge, dont la période mérovingienne est la plus représentée, a été attestée sur toute l'emprise. Considérant les découvertes faites pour cette période (notamment un four de potier et des indices concernant la métallurgie des non-ferreux), il se peut qu'il s'agisse d'un secteur artisanal au sein d'un habitat

Chamigny (Seine-et-Marne), Rue de la Marne - RD 80 : lieu-dit "La Grande Maison" : rapport de diagnostic

Un projet d'aménagement de lotissement déposé par la société Nexity sur la commune de Chamigny a conduit à la prescription d'un diagnostic archéologique par le Service régional d'Archéologie (DRAC, Île-de-France).Portant sur une surface totale prescrite de 22 044 m2, ce diagnostic a été réalisé par l'Inrap de mi-septembre à début octobre 2015.10,79 % de l'emprise prescrite ont été sondés sous forme de tranchées, de sondages profonds et d'ouvertures complémentaires. Ce diagnostic a permis la mise au jour de quatre sites chronologiques distincts. En premier, lieu, un niveau paléolithique moyen a été découvert lors de l'ouverture des sondages profonds 2 et 3, à une profondeur de 2,75 m et sur une épaisseur de 0,25 m. Ensuite, la présence d'une fosse datée du Néolithique ancien et plus particulièrement de la culture Blicquy-Villeneuve-Saint-Germain indique la présence d'une occupation de type habitat. Une sépulture collective de 5 m de long sur 3 m de large, dont l'état de conservation parait médiocre, dont est au moins attestée à deux étapes différentes du Néolithique final (IIIe millénaire avant J.-C.) avec une réutilisation à l'Epi-Campaniforme, a été mise au jour.Enfin, une très forte présence du haut Moyen Âge, dont la période mérovingienne est la plus représentée, a été attestée sur toute l'emprise. Considérant les découvertes faites pour cette période (notamment un four de potier et des indices concernant la métallurgie des non-ferreux), il se peut qu'il s'agisse d'un secteur artisanal au sein d'un habitat

The effect of hyperoxia on inflammation and platelet responses in an ex vivo extracorporeal membrane oxygenation circuit

Use of extracorporeal membrane oxygenation (ECMO) is expanding, however, it is still associated with significant morbidity and mortality. Activation of inflammatory and innate immune responses and hemostatic alterations contribute to complications. Hyperoxia may play a role in exacerbating these responses. Nine ex vivo ECMO circuits were tested using fresh healthy human whole blood, with two oxygen levels: 21% inspired fraction of oxygen (FiO(2); mild hyperoxia; n = 5) and 100% FiO(2)(severe hyperoxia; n = 4). Serial blood samples were taken for analysis of platelet aggregometry, leukocyte activation, inflammatory, and oxidative stress markers. ECMO resulted in reduced adenosine diphosphate- (P < .05) and thrombin receptor activating peptide-induced (P < .05) platelet aggregation, as well as increasing levels of the neutrophil activation marker, neutrophil elastase (P = .013). Additionally, levels of the inflammatory chemokine interleukin-8 were elevated (P < .05) and the activity of superoxide dismutase, a marker of oxidative stress, was increased (P = .002). Hyperoxia did not augment these responses, with no significant differences detected between mild and severe hyperoxia. Our ex vivo model of ECMO revealed that the circuit itself triggers a pro-inflammatory and oxidative stress response, however, exposure to supra-physiologic oxygen does not amplify that response. Extended-duration studies and inclusion of an endothelial component could be beneficial in characterizing longer term changes

An appraisal of lung computer tomography in very early anti-inflammatory treatment of two different ovine ARDS phenotypes

Abstract Mortality and morbidity of Acute Respiratory Distress Syndrome (ARDS) are largely unaltered. A possible new approach to treatment of ARDS is offered by the discovery of inflammatory subphenotypes. In an ovine model of ARDS phenotypes, matching key features of the human subphenotypes, we provide an imaging characterization using computer tomography (CT). Nine animals were randomized into (a) OA (oleic acid, hypoinflammatory; n = 5) and (b) OA-LPS (oleic acid and lipopolysaccharides, hyperinflammatory; n = 4). 48 h after ARDS induction and anti-inflammatory treatment, CT scans were performed at high (H) and then low (L) airway pressure. After CT, the animals were euthanized and lung tissue was collected. OA-LPS showed a higher air fraction and OA a higher tissue fraction, resulting in more normally aerated lungs in OA-LPS in contrast to more non-aerated lung in OA. The change in lung and air volume between H and L was more accentuated in OA-LPS, indicating a higher recruitment potential. Strain was higher in OA, indicating a higher level of lung damage, while the amount of lung edema and histological lung injury were largely comparable. Anti-inflammatory treatment might be beneficial in terms of overall ventilated lung portion and recruitment potential, especially in the OA-LPS group

Thyroid follicle development requires Smad1/Smad5- and endothelial-dependent basement membrane assembly

Thyroid follicles, the functional units of the thyroid gland, are delineated by a monolayer of thyrocytes resting on a continuous basement membrane. Developmental mechanisms whereby follicles are formed by reorganization of a non-structured mass of non-polarized epithelial cells (folliculogenesis) largely unknown. Here we show that assembly of the epithelial basement membrane is critical for folliculogenesis and is controlled by endothelial cell invasion and by BMP-Smad signaling in thyrocytes. Thyroid-specific double Smad1 and Smad5 knockout mice (Smad1/5(dKO)) displayed growth retardation, hypothyroidism and defective follicular architecture. In Smad1/5(dKO)embryonic thyroids, epithelial cells remained associated in large clusters and formed small follicles. Although similar follicular defects are found in Vegfa(KO)thyroids, Smad1/5(dKO)thyroids had normal endothelial cell density yet impaired endothelial differentiation. Interestingly, both Vegfa(KO)and Smad1/5(dKO)thyroids displayed impaired basement membrane assembly. Furthemore, conditioned medium (CM) from embryonic endothelial progenitor cells (eEPC) rescued the folliculogenic defects of both Smad1/5(dKO)and Vegfa(KO)thyroids. Laminin α1β1γ1, abundantly released by eEPC into CM, was critically required for folliculogenesis. Thus, epithelial Smad signaling and endothelial cell invasion promote folliculogenesis via assembly of the basement membrane

Thyroid follicle development requires Smad1/Smad5- and endothelial-dependent basement membrane assembly

Thyroid follicles, the functional units of the thyroid gland, are delineated by a monolayer of thyrocytes resting on a continuous basement membrane. Developmental mechanisms whereby follicles are formed by reorganization of a non-structured mass of non-polarized epithelial cells (folliculogenesis) largely unknown. Here we show that assembly of the epithelial basement membrane is critical for folliculogenesis and is controlled by endothelial cell invasion and by BMP-Smad signaling in thyrocytes. Thyroid-specific double Smad1 and Smad5 knockout mice (Smad1/5(dKO)) displayed growth retardation, hypothyroidism and defective follicular architecture. In Smad1/5(dKO)embryonic thyroids, epithelial cells remained associated in large clusters and formed small follicles. Although similar follicular defects are found in Vegfa(KO)thyroids, Smad1/5(dKO)thyroids had normal endothelial cell density yet impaired endothelial differentiation. Interestingly, both Vegfa(KO)and Smad1/5(dKO)thyroids displayed impaired basement membrane assembly. Furthemore, conditioned medium (CM) from embryonic endothelial progenitor cells (eEPC) rescued the folliculogenic defects of both Smad1/5(dKO)and Vegfa(KO)thyroids. Laminin α1β1γ1, abundantly released by eEPC into CM, was critically required for folliculogenesis. Thus, epithelial Smad signaling and endothelial cell invasion promote folliculogenesis via assembly of the basement membrane.status: publishe