Role of endocrine status and cell type in adhesion of human endometrial cells to the peritoneum in nude mice

Objective: To investigate the role of different cellular types (epithelial and stromal endometrial cells and peritoneal cells) in the ectopic implantation of endometrium and to evaluate the importance of endocrine environment on the adhesion of endometrial cells to the peritoneum. Design: Experimental prospective study. Setting: University hospital, department of cell biology. Animal(s): One hundred one nude mice. Intervention(s): Monolayer culture of human epithelial and stromal endometrial cells obtained from patients undergoing hysterectomy or laparoscopy for benign disease. Intraperitoneal injection of cells into nude mice. Main Outcome Measure(s): Two weeks after cell injection, adhesion of endometrial cells was evaluated by histological and immunohistochemical examination. Result(s): Mixed cultures of stromal and epithelial cells, but not purified epithelial or stromal cells alone, adhered to the mouse peritoneum and led to endometriotic-like nodules. Pretreatment of cells with estrogen alone or with estrogen and progestin resulted in a higher percentage of animals developing endometriotic-like nodules, whereas treatment with progestin alone did not affect endometriotic implantation. Conclusion(s): Our data indicate that the success of endometrial cell implantation is dependent on the cooperativeness between stromal and epithelial endometrial cells, as well as on the endocrine environment of endometrial cells, but not that of recipient animals. The results emphasize the role of both endometrial cell types for ectopic implantation. (C) 2002 by American Society for Reproductive Medicine.Peer reviewe

Scoring system for renal pathology in Fabry disease: report of the International Study Group of Fabry Nephropathy (ISGFN)

Background. In Fabry nephropathy, alpha-galactosidase deficiency leads to accumulation of glycosphingolipids in all kidney cell types, proteinuria and progressive loss of kidney function. Methods. An international working group of nephrologists from 11 Fabry centres identified adult Fabry patients, and pathologists scored histologic changes on renal biopsies. A standardized scoring system was developed with a modified Delphi technique assessing 59 Fabry nephropathy cases. Each case was scored independently of clinical information by at least three pathologists with an average final score reported. Results. We assessed 35 males (mean age 36.4 years) and 24 females (43.9 years) who mostly had clinically mild Fabry nephropathy. The average serum creatinine was 1.3mg/dl (114.9 μmol/l); estimated glomerular filtration rate was 81.7 ml/min/1.73 m2 and urine protein to creatinine ratio was 1.08 g/g (122.0 mg/mmol). Males had greater podocyte vacuolization on light microscopy (mean score) and glycosphingolipid inclusions on semi-thin sections than females. Males also had significantly more proximal tubule, peritubular capillary and vascular intimal inclusions. Arteriolar hyalinosis was similar, but females had significantly more arterial hyalinosis. Chronic kidney disease stage correlated with arterial and glomerular sclerosis scores. Significant changes, including segmental and global sclerosis, and interstitial fibrosis were seen even in patients with stage 1-2 chronic kidney disease with minimal proteinuria. Conclusions. The development of a standardized scoring system of both disease-specific lesions, i.e. lipid deposition related, and general lesions of progression, i.e. fibrosis and sclerosis, showed a spectrum of histologic appearances even in early clinical stage of Fabry nephropathy. These findings support the role of kidney biopsy in the baseline evaluation of Fabry nephropathy, even with mild clinical disease. The scoring system will be useful for longitudinal assessment of prognosis and responses to therapy for Fabry nephropath

Extensive blood transcriptome analysis reveals cellular signaling networks activated by circulating glycocalyx components reflecting vascular injury in COVID-19

BackgroundDegradation of the endothelial protective glycocalyx layer during COVID-19 infection leads to shedding of major glycocalyx components. These circulating proteins and their degradation products may feedback on immune and endothelial cells and activate molecular signaling cascades in COVID-19 associated microvascular injury. To test this hypothesis, we measured plasma glycocalyx components in patients with SARS-CoV-2 infection of variable disease severity and identified molecular signaling networks activated by glycocalyx components in immune and endothelial cells.MethodsWe studied patients with RT-PCR confirmed COVID-19 pneumonia, patients with COVID-19 Acute Respiratory Distress Syndrome (ARDS) and healthy controls (wildtype, n=20 in each group) and measured syndecan-1, heparan sulfate and hyaluronic acid. The in-silico construction of signaling networks was based on RNA sequencing (RNAseq) of mRNA transcripts derived from blood cells and of miRNAs isolated from extracellular vesicles from the identical cohort. Differentially regulated RNAs between groups were identified by gene expression analysis. Both RNAseq data sets were used for network construction of circulating glycosaminoglycans focusing on immune and endothelial cells.ResultsPlasma concentrations of glycocalyx components were highest in COVID-19 ARDS. Hyaluronic acid plasma levels in patients admitted with COVID-19 pneumonia who later developed ARDS during hospital treatment (n=8) were significantly higher at hospital admission than in patients with an early recovery. RNAseq identified hyaluronic acid as an upregulator of TLR4 in pneumonia and ARDS. In COVID-19 ARDS, syndecan-1 increased IL-6, which was significantly higher than in pneumonia. In ARDS, hyaluronic acid activated NRP1, a co-receptor of activated VEGFA, which is associated with pulmonary vascular hyperpermeability and interacted with VCAN (upregulated), a proteoglycan important for chemokine communication.ConclusionsCirculating glycocalyx components in COVID-19 have distinct biologic feedback effects on immune and endothelial cells and result in upregulation of key regulatory transcripts leading to further immune activation and more severe systemic inflammation. These consequences are most pronounced during the early hospital phase of COVID-19 before pulmonary failure develops. Elevated levels of circulating glycocalyx components may early identify patients at risk for microvascular injury and ARDS. The timely inhibition of glycocalyx degradation could provide a novel therapeutic approach to prevent the development of ARDS in COVID-19

Progranulin signaling in sepsis, community-acquired bacterial pneumonia and COVID-19: a comparative, observational study

BACKGROUND Progranulin is a widely expressed pleiotropic growth factor with a central regulatory effect during the early immune response in sepsis. Progranulin signaling has not been systematically studied and compared between sepsis, community-acquired pneumonia (CAP), COVID-19 pneumonia and a sterile systemic inflammatory response (SIRS). We delineated molecular networks of progranulin signaling by next-generation sequencing (NGS), determined progranulin plasma concentrations and quantified the diagnostic performance of progranulin to differentiate between the above-mentioned disorders using the established biomarkers procalcitonin (PCT), interleukin-6 (IL-6) and C-reactive protein (CRP) for comparison. METHODS The diagnostic performance of progranulin was operationalized by calculating AUC and ROC statistics for progranulin and established biomarkers in 241 patients with sepsis, 182 patients with SIRS, 53 patients with CAP, 22 patients with COVID-19 pneumonia and 53 healthy volunteers. miRNAs and mRNAs in blood cells from sepsis patients (n = 7) were characterized by NGS and validated by RT-qPCR in an independent cohort (n = 39) to identify canonical gene networks associated with upregulated progranulin at sepsis onset. RESULTS Plasma concentrations of progranulin (ELISA) in patients with sepsis were 57.5 (42.8-84.9, Q25-Q75) ng/ml and significantly higher than in CAP (38.0, 33.5-41.0~ng/ml, p < 0.001), SIRS (29.0, 25.0-35.0~ng/ml, p < 0.001) and the healthy state (28.7, 25.5-31.7~ng/ml, p < 0.001). Patients with COVID-19 had significantly higher progranulin concentrations than patients with CAP (67.6, 56.6-96.0 vs. 38.0, 33.5-41.0~ng/ml, p < 0.001). The diagnostic performance of progranulin for the differentiation between sepsis vs. SIRS (n = 423) was comparable to that of procalcitonin. AUC was 0.90 (95% CI = 0.87-0.93) for progranulin and 0.92 (CI = 0.88-0.96, p = 0.323) for procalcitonin. Progranulin showed high discriminative power to differentiate bacterial CAP from COVID-19 (sensitivity 0.91, specificity 0.94, AUC 0.91 (CI = 0.8-1.0) and performed significantly better than PCT, IL-6 and CRP. NGS and partial RT-qPCR confirmation revealed a transcriptomic network of immune cells with upregulated progranulin and sortilin transcripts as well as toll-like-receptor 4 and tumor-protein 53, regulated by miR-16 and others. CONCLUSIONS Progranulin signaling is elevated during the early antimicrobial response in sepsis and differs significantly between sepsis, CAP, COVID-19 and SIRS. This suggests that progranulin may serve as a novel indicator for the differentiation between these disorders. TRIAL REGISTRATION Clinicaltrials.gov registration number NCT03280576 Registered November 19, 2015

The evolving SARS-CoV-2 epidemic in Africa: Insights from rapidly expanding genomic surveillance.

Investment in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequencing in Africa over the past year has led to a major increase in the number of sequences that have been generated and used to track the pandemic on the continent, a number that now exceeds 100,000 genomes. Our results show an increase in the number of African countries that are able to sequence domestically and highlight that local sequencing enables faster turnaround times and more-regular routine surveillance. Despite limitations of low testing proportions, findings from this genomic surveillance study underscore the heterogeneous nature of the pandemic and illuminate the distinct dispersal dynamics of variants of concern-particularly Alpha, Beta, Delta, and Omicron-on the continent. Sustained investment for diagnostics and genomic surveillance in Africa is needed as the virus continues to evolve while the continent faces many emerging and reemerging infectious disease threats. These investments are crucial for pandemic preparedness and response and will serve the health of the continent well into the 21st century

The evolving SARS-CoV-2 epidemic in Africa: Insights from rapidly expanding genomic surveillance

INTRODUCTION Investment in Africa over the past year with regard to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequencing has led to a massive increase in the number of sequences, which, to date, exceeds 100,000 sequences generated to track the pandemic on the continent. These sequences have profoundly affected how public health officials in Africa have navigated the COVID-19 pandemic. RATIONALE We demonstrate how the first 100,000 SARS-CoV-2 sequences from Africa have helped monitor the epidemic on the continent, how genomic surveillance expanded over the course of the pandemic, and how we adapted our sequencing methods to deal with an evolving virus. Finally, we also examine how viral lineages have spread across the continent in a phylogeographic framework to gain insights into the underlying temporal and spatial transmission dynamics for several variants of concern (VOCs). RESULTS Our results indicate that the number of countries in Africa that can sequence the virus within their own borders is growing and that this is coupled with a shorter turnaround time from the time of sampling to sequence submission. Ongoing evolution necessitated the continual updating of primer sets, and, as a result, eight primer sets were designed in tandem with viral evolution and used to ensure effective sequencing of the virus. The pandemic unfolded through multiple waves of infection that were each driven by distinct genetic lineages, with B.1-like ancestral strains associated with the first pandemic wave of infections in 2020. Successive waves on the continent were fueled by different VOCs, with Alpha and Beta cocirculating in distinct spatial patterns during the second wave and Delta and Omicron affecting the whole continent during the third and fourth waves, respectively. Phylogeographic reconstruction points toward distinct differences in viral importation and exportation patterns associated with the Alpha, Beta, Delta, and Omicron variants and subvariants, when considering both Africa versus the rest of the world and viral dissemination within the continent. Our epidemiological and phylogenetic inferences therefore underscore the heterogeneous nature of the pandemic on the continent and highlight key insights and challenges, for instance, recognizing the limitations of low testing proportions. We also highlight the early warning capacity that genomic surveillance in Africa has had for the rest of the world with the detection of new lineages and variants, the most recent being the characterization of various Omicron subvariants. CONCLUSION Sustained investment for diagnostics and genomic surveillance in Africa is needed as the virus continues to evolve. This is important not only to help combat SARS-CoV-2 on the continent but also because it can be used as a platform to help address the many emerging and reemerging infectious disease threats in Africa. In particular, capacity building for local sequencing within countries or within the continent should be prioritized because this is generally associated with shorter turnaround times, providing the most benefit to local public health authorities tasked with pandemic response and mitigation and allowing for the fastest reaction to localized outbreaks. These investments are crucial for pandemic preparedness and response and will serve the health of the continent well into the 21st century

Role of Zr and Ga additions on the hydrogen process of Nd-Fe-B magnets

International audienceHighly coercive Nd-Fe-B powders for bonded magnets have been successfully produced applying hydrogen processes such as hydrogen decrepitation (HD) or hydrogen-disproportionation-dehydrogenation-recombination (HDDR). XAFS experiments were performed in order to localise the extra elements. Zr addition results in the formation of fine ZrB2 precipitates, whereas Ga substitutes the Fe(4c) site in the Nd2Fe14B structure. Systematic analyses of the hydrogen disproportionation kinetics by thermomagnetic measurements have permitted to point out different regimes of reactivity depending on both the used extra elements and the temperature. It is found that the ZrB2 precipitates act as barriers limiting hydrogen diffusion at the crystallites boundaries and lead to uncompleted disproportionation reactions. Then the Nd2Fe14B remaining particles act as nucleation centres that can be used to induce oriented growth during the recombination step

Belgique

INTRODUCTION | Renal ischemia/reperfusion (I/R) is the leading cause of acute kidney injury (AKI). Ischemic preconditioning (IPC) may help to attenuate the severity of I/Rassociated AKI. Whole-body irradiation induces renal IPC in mice, although the pathways remain largely unknown. Furthermore, the impact of kidney-centred irradiation on renal resistance against I/R has not been studied. First, we comprehensively investigate the pathways involved in renal irradiation. Next, we assess the functional impact of renal irradiation applied I/R injury. Finally, we test whether Sunitinib-mediated inhibition of the angiogenesis prevents irradiation-associated IPC. METHODS | Exp1: Renal irradiation (8.5Gy) was performed in male C57bl/6 mice(n=10). One month later, total kidney RNA was extracted from irradiated and control (n=5) mice for comparative RNA-Seq. Exp2: After renal irradiation, the right kidneys were removed, and the left kidneys undergo ischemia(30min)/reperfusion(48h) at Days 7-14-28 post irradiation(n=8). Exp3: Following the same protocol of I/R at Day14, 3 groups were compared(n=8): 1/irradiation; 2/irradiation and gavage with Sunitinib from Day2 to 13; 3/control group without irradiation or gavage. RESULTS | Exp1: RNAseq showen up-regulation of angiogenesis signalling pathways. Expressions of angiogenesis markers (CD31, VEGF) showed an increase at both mRNA and protein levels in irradiated kidneys (p<0.01). Exp2: Following I/R, Blood Urea Nitrogen (BUN) and Creatinine (SCr) levels were lower in the irradiated mice compared to controls: (BUN: 86.2±6.8 vs. 454.5±27.2mg/dl; SCr: 0.1±0.01 vs. 1.7±0.2mg/dl, p<0.01). The renal infiltration by CD11b-(187±32 vs 477±20/mm2) and F4-80-positive cells (110±22 vs 212±25/mm2) was reduced in the irradiated group. VEGF and CD31 expression was increased in irradiated kidneys at both mRNA and protein levels(p<0,01). Exp3: One-way analysis of variance followed by Tukey’s test showed that, following I/R, BUN and SCr levels were lower in the irradiated group compared to controls (BUN: 106.1±33.6 vs. 352.2±54.3mg/dl; SCr: 0.3±0.13 vs. 1±0.2mg/dl), and in irradiated group compared to the irradiated-exposed group to Sunitinib (BUN: 106.1±33.6 vs. 408.4±54.9mg/dl; SCr: 0.3±0.12 vs. 1.5±0.3mg/dl; p<0.01). CONCLUSION | Renal irradiation induces the activation of angiogenesis in mice. Renal irradiation leads to IPC, with preserved renal function and attenuated inflammation post I/R. Exposure to the anti-angiogenic drug Sunitinib post-irradiation prevents the irradiation-induced IPC.Impact de l'irradiation dans le préconditionnement ischémique réna

Belgique

Introduction L’irradiation corporelle induit un conditionnement ischémique rénal(CIR) chez la souris. Les mécanismes cellulaires sont méconnus, hormis un possible rôle de la néo-angiogenèse. Description claire et complète de l'expérience Dans cette étude, nous étudions les voies impliquées dans l'irradiation rénale. Ensuite, nous analysons l'impact fonctionnel sur les reins avant ischémie rénale/reperfusion(I/R). Enfin, nous testons si l'inhibition de l'angiogenèse par le Sunitinib empêche le CIR associé à l'irradiation. Méthodes (une explication détaillée de votre analyse est attendue) Exp1. Une irradiation rénale(8Gy) est réalisée chez des C57bl/6 mâles(n=10). Un mois plus tard, l'ARN rénal total est extrait pour un RNAseq comparatif. Exp2. À 7- 14-28 jours post irradiation rénale, les reins droits sont néphrectomisés et les reins gauches subissent une ischémie(30min)/reperfusion(48h) (n=8/timing). Exp3. Suivant le même protocole d'I/R à J14, 3 groupes sont comparés(n=8/groupe) : 1/ irradiation ; 2 /irradiation et gavage au Sunitinib de J2 à J13 ; 3/ groupe témoin sans irradiation ni gavage. Résultats obtenus ou attendus Exp1. RNAseq montre une up-régulation des voies de l'angiogenèse. L’expression de VEGF et CD31 est significativement augmentée au niveau ARNm et protéique dans les reins irradiés. Exp2. Après I/R à J14 post irradiation, les taux sériques d’urée(BUN) et de créatinine(SCr) sont plus faibles chez les souris irradiées par rapport aux témoins(BUN : 86,2±6,8 vs 454,5±27,2 mg/dl ; SCr: 0,1±0,01 vs 1,7±0,2 mg/dl, p<0,01). L'infiltration rénale par les macrophages CD11b(187±32 vs 477±20/mm²) et F4-80(110±22 vs 212±25/mm²) est significativement moindre dans le groupe irradié. L'expression de VEGF et CD31 est majorée dans les reins irradiés à partir de J14. Exp3. Après I/R, les taux sériques de BUN et de SCr dans le groupe irradié-exposé au Sunitinib sont similaires au groupe contrôle(BUN 352,2±54,3 vs. 408,4 ± 54,9 mg/dl ; SCr : 1,5±0,3 vs. 1±0,2 mg/dl). Conclusion L'irradiation rénale induit l'activation de l'angiogenèse chez la souris et est associée à un CIR, avec fonction rénale préservée et inflammation atténuée post I/R. L'exposition au Sunitinib post-irradiation empêche ce CIR.Impact de l'irradiation dans le préconditionnement ischémique réna

Belgique

Background: Whole-body irradiation induces renal ischemic preconditioning (RIP) in mice, possibly via neoangiogenesis. Here, we test whether Sunitinib-mediated inhibition of angiogenesis prevents the irradiation-associated RIP. ● Methods: After kidney-centred irradiation (8.56 Gy), the right kidneys were removed and harvested, and the left kidneys underwent ischemia (30min) / reperfusion (48h) (I/R) at Day 14. Three groups were compared (n=8/group): 1/irradiation; 2/irradiation and gavage with Sunitinib (40 mg/kg) from D2 to D13; 3/control group without irradiation or gavage. Renal sections from the 3 groups post-I/R were stained by Periodic Acid Schiff (PAS). I/R-associated acute tubular necrosis was blindly evaluated by a renal pathologist using the histological Jablonski score. The expression of inflammatory markers CD11b and F4/80 was comparatively quantified by immunostaining. The expression of vascular markers CD31 and VEGF in nonischemic kidneys were quantified by real-time qPCR. ● Results: One-way analysis of variance followed by Tukey’s test showed that, following I/R, serum levels of urea (BUN) and creatinine (SCr) were significantly lower in pre-irradiated mice compared to controls (BUN: 106.1±33.6 vs. 352.2±54.3mg/dl; SCr: 0.3±0.13 vs. 1±0.2mg/dl), as well as in pre-irradiated mice compared to the irradiated mice fed with Sunitinib (BUN: 106.1±33.6 vs. 408.4±54.9mg/dl; SCr: 0.3±0.12 vs. 1.5±0.3mg/dl). No difference was observed between the Sunitinib group and the control group. Jablonski’s severity score was lower in pre-irradiated mice compared to control group and Sunitinib group (p<0.01). The renal infiltration by CD11b- (560±32 vs. 308±21/mm²) and F4-80 positive cells (430±35 vs. 312±19/mm²) was significantly reduced in the irradiated group compared to controls. At mRNA levels, the renal expression of VEGF and CD31 was increased in the irradiated group but not in the Sunitinib group (p<0.01). ● Conclusions: Renal irradiation before I/R is associated with preserved renal function and attenuated inflammation post I/R. Sunitinib administration prevents the irradiation-induced RIP.Impact de l'irradiation dans le préconditionnement ischémique réna