Damage-Associated Molecular Patterns and Pattern Recognition Receptors in the Podocyte.

ABSTRACT: Podocytes possess immune system components allowing for a variety of innate responses to endogenous and exogenous stimuli. Recently, several groups have linked inappropriate innate immune signaling to podocyte injury, particularly chronic, sustained injury; however, the immune capabilities of podocytes have not been fully elucidated. Damage associated molecular patterns (DAMPs) are endogenous danger molecules released from damaged cells, including podocytes, and can elicit an inflammatory response and recruit immune cells to areas of injury. This is done through binding to pattern recognition receptors (PRR). Thought largely to be protective and responsive to injury or infection, recent evidence suggests signaling via DAMP pathways can aggravate and promote chronic diseases already associated with inflammation. The purpose of this narrative review is to highlight current knowledge with respect to specific podocyte DAMPs and PRRs, and to provide insight into ongoing work and possible future research avenues to advance our understanding of podocyte immune mechanisms.

Retrospective evaluation of a right heart parameter score in the assessment of precapillary pulmonary hypertension in dogs (2017-2021): 135 cases.

OBJECTIVE: To determine if emergency and critical care residents can identify moderate to severe precapillary pulmonary hypertension on cardiologist-obtained cineloops using a pulmonary hypertension score (PHS) and report the interobserver variability of the PHS. DESIGN: Multicenter, retrospective, case-control study from 2017 to 2021. SETTING: Private referral center and veterinary teaching hospital. ANIMALS: One hundred and thirty-five client-owned dogs that underwent diagnostic echocardiography. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Medical records of dogs with stage B1 myxomatous mitral valve disease (MMVD) and dogs diagnosed with precapillary pulmonary hypertension (PCPH) via echocardiograms were reviewed. Dogs were categorized by a cardiologist into 5 groups (normal, B1 MMVD, mild, moderate, and severe PCPH) based on Doppler pulmonary pressure gradients and right heart morphology. Cineloops from each case were subjectively evaluated by emergency and critical care residents for the presence of right atrial and ventricular enlargement, right ventricular hypertrophy, interventricular septal flattening, and pulmonary artery and trunk enlargement to form a composite pulmonary hypertension score out of 8 (PHS8). When available, signs of peritoneal effusion and distention of the caudal vena cava were subjectively assessed to generate a pulmonary hypertension score out of 10 (PHS10). There was excellent discrimination of moderate to severe PCPH versus grouped absent to mild PCPH using PHS8 (area under the receiver operator curve [AUC] [95% confidence interval, CI] = 0.90 [0.84-0.95], P < 0.0001) and PHS10 (AUC [95% CI] = 0.89 [0.81-0.97], P < 0.0001). PHS8 ≥3 was 64% sensitive and 98% specific for moderate to severe PCPH (positive likelihood ratio [LR+] 32, negative likelihood ration [LR-] 0.37). PHS10 ≥ 3.3 was 64% sensitive and 92% specific for moderate to severe PCPH (LR+ 8, LR- 0.39). Interobserver agreement was good to excellent (intraclass correlation coefficient [ICC] = 0.74 [95% CI: 0.66-0.80], n = 135). CONCLUSIONS: Residents identified moderate to severe PCPH in dogs using PHS on cineloops previously obtained by a cardiologist. The interrater agreement was good to excellent with limited training. Prospective studies to determine if residents can obtain diagnostic images for PHS are warranted.

Endothelial NOX5 Obliterates the Reno-Protective Effect of Nox4 Deletion by Promoting Renal Fibrosis via Activation of EMT and ROS-Sensitive Pathways in Diabetes.

Chronic hyperglycemia induces intrarenal oxidative stress due to the excessive production of reactive oxygen species (ROS), leading to a cascade of events that contribute to the development and progression of diabetic kidney disease (DKD). NOX5, a pro-oxidant NADPH oxidase isoform, has been identified as a significant contributor to renal ROS in humans. Elevated levels of renal ROS contribute to endothelial cell dysfunction and associated inflammation, causing increased endothelial permeability, which can disrupt the renal ecosystem, leading to progressive albuminuria and renal fibrosis in DKD. This study specifically examines the contribution of endothelial cell-specific human NOX5 expression in renal pathology in a transgenic mouse model of DKD. This study additionally compares NOX5 with the previously characterized NADPH oxidase, NOX4, in terms of their relative roles in DKD. Regardless of NOX4 pathway, this study found that endothelial cell-specific expression of NOX5 exacerbates renal injury, albuminuria and fibrosis. This is attributed to the activation of the endothelial mesenchymal transition (EMT) pathway via enhanced ROS formation and the modulation of redox-sensitive factors. These findings underscore the potential therapeutic significance of NOX5 inhibition in human DKD. The study proposes that inhibiting NOX5 could be a promising approach for mitigating the progression of DKD and strengthens the case for the development of NOX5-specific inhibitors as a potential therapeutic intervention.

Left atrial measurement in lateral versus sternal recumbency in cats undergoing focused cardiac ultrasound examination.

Objective: To compare left atrial measurements carried out by an emergency and critical care (ECC) clinician on cats in lateral and sternal recumbency. Animals and procedures: A prospective observational study was conducted between December 2019 and January 2021 at the university teaching hospital at University of Liège. One hundred and two hospitalized cats were enrolled. Focused cardiac ultrasound (FOCUS) was performed in right lateral and sternal recumbency by a single FOCUS-trained ECC resident. Standard right parasternal long- and short-axis views were recorded. After randomization of the cineloops, the same blinded resident measured maximal left atrial dimension (LAD) and the ratio of left atrial to aortic diameter (LA:Ao). Reproducibility was assessed using the Bland-Altman method. Results: The LA:Ao and LAD measurements in lateral (LA:Ao median: 1.37, range: 1.02 to 3.22; LAD median: 13.25, range: 7.90 to 32.90) and sternal (LA:Ao median: 1.38, range: 1.06 to 3.22; LAD median: 13.00, range: 8.00 to 32.90) recumbency were not significantly different (bias: -0.003, CI -0.014, 0.007; and bias: -0.101, CI -0.231, 0.029, respectively). Conclusions and clinical relevance: The FOCUS technique was successfully applied in sternal recumbency in almost all cats. The LAD and LA:Ao measured in sternal and lateral recumbency were not significantly different. Cardiac left atrial measurements obtained using FOCUS can be reliably assessed in sternal recumbency in hospitalized, stable cats.

Mesure de l'oreillette gauche en décubitus latéral versus sternal chez les chats soumis à une échographie cardiaque focalisée. Objectif: Comparer les mesures de l'oreillette gauche effectuées par un clinicien des urgences et soins intensifs (ECC) sur des chats en décubitus latéral et sternal. Animaux et procédures: Une étude observationnelle prospective a été menée entre décembre 2019 et janvier 2021 au CHU de l'Université de Liège. Cent deux chats hospitalisés ont été enrôlés. L'échographie cardiaque focalisée (FOCUS) a été réalisée en décubitus latéral droit et sternal par un seul résident ECC formé au FOCUS. Des vues parasternales droites grand et petit axe standards ont été enregistrées. Après randomisation des cineloops, le même résident en aveugle a mesuré la dimension auriculaire gauche maximale (LAD) et le rapport entre le diamètre de l'oreillette gauche et celui de l'aorte (LA:Ao). La reproductibilité a été évaluée à l'aide de la méthode de Bland-Altman. Résultats: Les mesures LA:Ao et LAD en décubitus latéral (LA:Ao médian : 1,37, intervalle : 1,02 à 3,22; LAD médian : 13,25, intervalle : 7,90 à 32,90) et sternal (LA:Ao médian : 1,38, intervalle : 1,06 à 3,22; médiane LAD : 13,00, intervalle : 8,00 à 32,90) n'étaient pas significativement différents (biais : −0,003, IC −0,014, 0,007; et biais : −0,101, IC −0,231, 0,029, respectivement). Conclusions et pertinence clinique: La technique FOCUS a été appliquée avec succès en décubitus sternal chez presque tous les chats. Le LAD et LA:Ao mesurés en décubitus sternal et latéral n'étaient pas significativement différents. Les mesures de l'oreillette cardiaque gauche obtenues à l'aide de FOCUS peuvent être évaluées de manière fiable en décubitus sternal chez les chats hospitalisés et stables.(Traduit par Dr Serge Messier).

The Proteome of Circulating Large Extracellular Vesicles in Diabetes and Hypertension.

Hypertension and diabetes induce vascular injury through processes that are not fully understood. Changes in extracellular vesicle (EV) composition could provide novel insights. Here, we examined the protein composition of circulating EVs from hypertensive, diabetic and healthy mice. EVs were isolated from transgenic mice overexpressing human renin in the liver (TtRhRen, hypertensive), OVE26 type 1 diabetic mice and wild-type (WT) mice. Protein content was analyzed using liquid chromatography-mass spectrometry. We identified 544 independent proteins, of which 408 were found in all groups, 34 were exclusive to WT, 16 were exclusive to OVE26 and 5 were exclusive to TTRhRen mice. Amongst the differentially expressed proteins, haptoglobin (HPT) was upregulated and ankyrin-1 (ANK1) was downregulated in OVE26 and TtRhRen mice compared with WT controls. Conversely, TSP4 and Co3A1 were upregulated and SAA4 was downregulated exclusively in diabetic mice; and PPN was upregulated and SPTB1 and SPTA1 were downregulated in hypertensive mice, compared to WT mice. Ingenuity pathway analysis identified enrichment in proteins associated with SNARE signaling, the complement system and NAD homeostasis in EVs from diabetic mice. Conversely, in EVs from hypertensive mice, there was enrichment in semaphroin and Rho signaling. Further analysis of these changes may improve understanding of vascular injury in hypertension and diabetes.

High fat diet is protective against kidney injury in hypertensive-diabetic mice, but leads to liver injury.

Chronic kidney disease (CKD) is a worldwide health burden with increases risk of end-stage renal function if left untreated. CKD induced in the context of metabolic syndrome (MS) increases risks of hypertension, hyperglycemia, excess body fat and dyslipidemia. To test if combining a high-fat diet (HFD) regimen onto the hypertensive/ diabetic phenotype would mimic features of MS induced-CKD in mice, hyperglycemia was induced in genetically hypertensive mice (Lin), followed by HFD regimen. For that, 8-week-old male were subjected to streptozotocin (STZ) intraperitoneal (i.p.) injections (50 mg/kg, 5 days consecutive). LinSTZ were fed a 60% kCal HFD for 8 weeks. Lin mice treated with STZ developed polydipsia, became hypertensive and hyperglycemic. HFD induced weight gain, protected against glomerular hypertrophy, scarring, and albuminuria at endpoint compared to regular diet fed LinSTZ. On the other hand, HFD induced steatosis, liver fibrosis, inflammation, and increase in AST/ALT ratio, characteristics of non-alcoholic liver disease. Taken together, our results show that LinSTZ mice fed a HFD did not lead to a more robust model of MS-induced CKD, protected against kidney injury, but inducing liver damage. More studies are necessary to understand the kidney protective mechanisms of HFD when superimposed with hypertension and type 1 diabetes.

Parkin coregulates glutathione metabolism in adult mammalian brain.

We recently discovered that the expression of PRKN, a young-onset Parkinson disease-linked gene, confers redox homeostasis. To further examine the protective effects of parkin in an oxidative stress model, we first combined the loss of prkn with Sod2 haploinsufficiency in mice. Although adult prkn-/-//Sod2± animals did not develop dopamine cell loss in the S. nigra, they had more reactive oxidative species and a higher concentration of carbonylated proteins in the brain; bi-genic mice also showed a trend for more nitrotyrosinated proteins. Because these redox changes were seen in the cytosol rather than mitochondria, we next explored the thiol network in the context of PRKN expression. We detected a parkin deficiency-associated increase in the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) in murine brain, PRKN-linked human cortex and several cell models. This shift resulted from enhanced recycling of GSSG back to GSH via upregulated glutathione reductase activity; it also correlated with altered activities of redox-sensitive enzymes in mitochondria isolated from mouse brain (e.g., aconitase-2; creatine kinase). Intriguingly, human parkin itself showed glutathione-recycling activity in vitro and in cells: For each GSSG dipeptide encountered, parkin regenerated one GSH molecule and was S-glutathionylated by the other (GSSG + P-SH [Formula: see text] GSH + P-S-SG), including at cysteines 59, 95 and 377. Moreover, parkin's S-glutathionylation was reversible by glutaredoxin activity. In summary, we found that PRKN gene expression contributes to the network of available thiols in the cell, including by parkin's participation in glutathione recycling, which involves a reversible, posttranslational modification at select cysteines. Further, parkin's impact on redox homeostasis in the cytosol can affect enzyme activities elsewhere, such as in mitochondria. We posit that antioxidant functions of parkin may explain many of its previously described, protective effects in vertebrates and invertebrates that are unrelated to E3 ligase activity.

Advancing Discovery Research in Nephrology in Canada: A Conference Report From the 2021 Molecules and Mechanisms Mediating Kidney Health and Disease (M3K) Scientific Meeting and Investigator Summit.

Purpose of conference: New discoveries arising from investigations into fundamental aspects of kidney development and function in health and disease are critical to advancing kidney care. Scientific meetings focused specifically on fundamental biology of the kidney can facilitate interactions, support the development of collaborative groups, and accelerate translation of key findings. The Canadian fundamental kidney researcher community has lacked such a forum. On December 3 to 4, 2021, the first Molecules and Mechanisms Mediating Kidney Health and Disease (M3K) Scientific Meeting and Investigator Summit was held to address this gap with the goal of advancing fundamental kidney research nationally. The meeting was held virtually and was supported by a planning and dissemination grant from the Canadian Institutes of Health Research. Attendees included PhD scientists, nephrology clinician scientists, engineers, industry representatives, graduate students, medical residents, and fellows. Sources of information: This report was prepared from the scientific program, registration numbers, and details obtained from the online platform WHOVA, and summaries written by organizers and participants of the 2021 meeting. Methods: A 21-person team, consisting of the organizing committee members and participants from the meeting, was assembled. Key highlights of the meeting and future directions were identified and the team jointly assembled this report. Key findings: Participation in the meeting was strong, with more than 140 attendees across a range of disciplines. The program featured state-of-the-art presentations on diabetic nephropathy, the immune system, kidney development, and fibrosis, and was heavily focused on trainee presentations. The moderated "Investigator Summit" identified key barriers to research advancement and discussed strategies for overcoming them. These included establishment of a pan-Canadian fundamental kidney research network, development of key resources, cross-pollination with clinical nephrology, better reintegration into the Canadian Society of Nephrology, and further establishment of identity and knowledge translation. Limitations and implications: The 2021 M3K meeting represented a key first step in uniting fundamental kidney researchers in Canada. However, it was universally agreed that regular meetings were necessary to sustain this momentum. The proceedings of this meeting and future actions to sustain the M3K Scientific Meeting and Investigator Summit are presented in this article.

Objectif de la conférence: De nouvelles découvertes découlant des enquêtes sur les aspects fondamentaux du développement et de la fonction des reins en santé ou malades sont essentielles pour faire progresser les soins rénaux. Les réunions scientifiques axées spécifiquement sur la biologie fondamentale du rein peuvent faciliter les interactions, appuyer le développement de groupes de collaboration et accélérer l'application des principaux résultats. La communauté canadienne des chercheurs fondamentaux en néphrologie a manqué d'un tel forum. Les 3 et 4 décembre 2021, le premier Sommet des chercheurs et la réunion scientifique M3K (Molecules and Mechanisms Mediating Kidney Health and Disease) sur les molécules et les médiateurs de la santé et des maladies rénales ont eu lieu pour combler cette lacune; l'objectif était de faire progresser la recherche fondamentale en néphrologie à l'échelle nationale. La réunion s'est tenue virtuellement et était financée par une subvention de planification et de diffusion des Instituts de recherche en santé du Canada. Des doctorants, cliniciens-chercheurs en néphrologie, ingénieurs, représentants de l'industrie, étudiants diplômés, résidents en médecine et en surspécialisation figuraient parmi les participants. Sources: Ce rapport a été préparé à partir du program scientifique, des informations et des numéros d'inscription tirés de la plateforme en ligne WHOVA, et des résumés rédigés par les organisateurs et les participants à la réunion de 2021. Méthodologie: Une équipe de 20 personnes composée de membres du comité organisateur et de participants à la réunion a été formée. Les principaux points saillants de la réunion et les orientations futures ont été déterminés, puis l'équipe a rédigé conjointement le présent rapport. Principaux résultats: La réunion s'est avérée un succès; plus de 140 personnes provenant d'un large éventail de disciplines y ont participé. Le program comprenait des présentations de pointe sur la néphropathie diabétique, le système immunitaire, le développement des reins et la fibrose, et était fortement axé sur des présentations par des stagiaires. Le « Sommet des chercheurs ¼, animé par un modérateur, a permis de déterminer les principaux obstacles à l'avancement de la recherche et de discuter des stratégies pour les surmonter. Ces dernières incluent notamment la création d'un réseau pancanadien de recherche fondamentale en néphrologie, le développement de ressources clés, la pollinisation croisée avec la néphrologie clinique, une « meilleure réintégration dans la Société canadienne de néphrologie ¼ et la poursuite de l'établissement de l'identité et de l'application des connaissances. Limites et implications: La réunion M3K de 2021 a constitué une première étape clé dans l'unification des chercheurs fondamentaux en néphrologie au Canada. On a cependant universellement convenu que des réunions régulières étaient nécessaires pour maintenir cet élan. Le compte rendu de cette réunion ainsi que les actions futures pour soutenir la réunion scientifique M3K et le Sommet des chercheurs sont présentés dans le présent article.

Septic pericardial disease and mediastinal abscessation in a cat with an intrathoracic needle foreign body.

OBJECTIVE: To describe the successful management of a cat with an intrathoracic sewing needle foreign body and septic pericardial effusion. CASE SUMMARY: A 10-year-old neutered female domestic longhair cat was referred for an intrathoracic metallic foreign body identified via thoracic radiography. Two weeks prior, the cat may have ingested a sewing needle. She was presented hemodynamically unstable; point-of-care thoracic ultrasound identified pericardial effusion with right atrial tamponade. Pericardiocentesis stabilized hemodynamic parameters. The effusion was grossly purulent, and bacterial culture grew an Actinomyces sp. The cat underwent a median sternotomy to remove the foreign body, debride associated mediastinal abscesses, and perform a partial pericardiectomy. Over the next 10 days, the cat was managed in hospital with a left unilateral thoracostomy tube and intermittent lavage of the pleural cavity. The cat was discharged, and follow-up at 7 days showed no evidence of pericardial or pleural effusion. NEW OR UNIQUE INFORMATION PROVIDED: Contrary to previous reports, this case shows that extra-gastrointestinal, specifically intrathoracic, sewing needle foreign bodies can cause significant morbidity in cats. To the authors' knowledge, this report is the first to describe septic pericardial disease resulting from documented foreign body ingestion in the cat. It is also the first case report of successful surgical management of mediastinal abscessation in the cat.

The use of caffeine as a respiratory stimulant in a cat.

OBJECTIVE: To report the use of caffeine as a respiratory stimulant in a cat with hypoventilation. CASE SUMMARY: A cat was mechanically ventilated due to persistent hypercapnia (Pet co2  > 75 mm Hg) following median sternotomy and thymectomy. After 3 days of mechanical ventilatory support, the cat would initiate breaths but failed weaning due to persistent hypercapnia. Following administration of intravenous caffeine (total 12 mg/kg over 24 h), respiratory and mental status rapidly improved. The cat was subsequently extubated and able to maintain Pvco2  < 50 mm Hg. The cat was later diagnosed with myasthenia gravis. Quality of life 13 months after discharge was reported as excellent by her owner. NEW OR UNIQUE INFORMATION PROVIDED: Caffeine may be considered as a respiratory stimulant in cats with hypoventilation.

The accuracy of tactile assessment of canine nose temperature to identify rectal hyperthermia and hypothermia in dogs presenting on an emergency basis.

The goals of this study were to evaluate whether touch can identify a warm nose as opposed to a cold nose, to examine the correlation between thermographically measured nose temperatures and rectal temperatures, and to calculate the accuracy of tactile assessment of nose temperature in detecting rectal hyperthermia and hypothermia in dogs. A total of 100 dogs presenting to an emergency room was prospectively enrolled. Tactile nose assessment was carried out on triage. Noses were subjectively categorized as warm, cold, or intermediate (neither warm nor cold). Thermographic nose temperatures were recorded using a thermal imaging camera. Tactile assessment categorized noses as warm, intermediate, or cold (P < 0.01). There was no correlation between thermographically measured nose temperature and rectal temperature (r = 0.02). Tactile assessment of noses as warm had a sensitivity of 29.4% and a specificity of 79.5% for detecting rectal hyperthermia; calculated test accuracy was 71%. Tactile assessment of noses as cold had a sensitivity of 54.5% and a specificity of 62.9%; calculated test accuracy was 62%. It was concluded that nose temperatures do not correlate with rectal temperatures. Tactile assessment of nose temperature is inaccurate for identifying rectal hyperthermia or hypothermia.

Les objectifs de cette étude étaient d'évaluer si le toucher peut identifier un nez chaud par opposition à un nez froid, d'examiner la corrélation entre les températures nasales mesurées thermographiquement et les températures rectales, et de calculer la précision de l'évaluation tactile de la température nasale dans la détection de l'hyperthermie et l'hypothermie rectale chez le chien. Un total de 100 chiens se présentant aux urgences a été enrôlé de manière prospective. Une évaluation tactile du nez a été réalisée lors du triage. Les nez ont été classés subjectivement comme chaud, froid ou intermédiaire (ni chaud ni froid). Les températures thermographiques du nez ont été enregistrées à l'aide d'une caméra thermique. L'évaluation tactile a classé les nez comme chauds, intermédiaires ou froids (P < 0,01). Il n'y avait pas de corrélation entre la température nasale mesurée par thermographie et la température rectale (r = 0,02). L'évaluation tactile des nez chauds avait une sensibilité de 29,4 % et une spécificité de 79,5 % pour détecter l'hyperthermie rectale; la précision calculée du test était de 71 %. L'évaluation tactile des nez froids avait une sensibilité de 54,5 % et une spécificité de 62,9 %; la précision calculée du test était de 62 %. Il a été conclu que les températures nasales ne sont pas corrélées avec les températures rectales. L'évaluation tactile de la température du nez est imprécise pour identifier l'hyperthermie ou l'hypothermie rectale.(Traduit par les auteurs).

Fatty acid mimetic PBI-4547 restores metabolic homeostasis via GPR84 in mice with non-alcoholic fatty liver disease.

Non-alcoholic Fatty Liver Disease (NAFLD) is the most common form of liver disease and is associated with metabolic dysregulation. Although G protein-coupled receptor 84 (GPR84) has been associated with inflammation, its role in metabolic regulation remains elusive. The aim of our study was to evaluate the potential of PBI-4547 for the treatment of NAFLD and to validate the role of its main target receptor, GPR84. We report that PBI-4547 is a fatty acid mimetic, acting concomitantly as a GPR84 antagonist and GPR40/GPR120 agonist. In a mouse model of diet-induced obesity, PBI-4547 treatment improved metabolic dysregulation, reduced hepatic steatosis, ballooning and NAFLD score. PBI-4547 stimulated fatty acid oxidation and induced gene expression of mitochondrial uncoupling proteins in the liver. Liver metabolomics revealed that PBI-4547 improved metabolic dysregulation induced by a high-fat diet regimen. In Gpr84-/- mice, PBI-4547 treatment failed to improve various key NAFLD-associated parameters, as was observed in wildtype littermates. Taken together, these results highlight a detrimental role for the GPR84 receptor in the context of meta-inflammation and suggest that GPR84 antagonism via PBI-4547 may reflect a novel treatment approach for NAFLD and its related complications.

Vascular contributions to 16p11.2 deletion autism syndrome modeled in mice.

While the neuronal underpinnings of autism spectrum disorder (ASD) are being unraveled, vascular contributions to ASD remain elusive. Here, we investigated postnatal cerebrovascular development in the 16p11.2df/+ mouse model of 16p11.2 deletion ASD syndrome. We discover that 16p11.2 hemizygosity leads to male-specific, endothelium-dependent structural and functional neurovascular abnormalities. In 16p11.2df/+ mice, endothelial dysfunction results in impaired cerebral angiogenesis at postnatal day 14, and in altered neurovascular coupling and cerebrovascular reactivity at postnatal day 50. Moreover, we show that there is defective angiogenesis in primary 16p11.2df/+ mouse brain endothelial cells and in induced-pluripotent-stem-cell-derived endothelial cells from human carriers of the 16p11.2 deletion. Finally, we find that mice with an endothelium-specific 16p11.2 deletion (16p11.2ΔEC) partially recapitulate some of the behavioral changes seen in 16p11.2 syndrome, specifically hyperactivity and impaired motor learning. By showing that developmental 16p11.2 haploinsufficiency from endothelial cells results in neurovascular and behavioral changes in adults, our results point to a potential role for endothelial impairment in ASD.

Nox (NADPH Oxidase) 1, Nox4, and Nox5 Promote Vascular Permeability and Neovascularization in Retinopathy.

Hypertension is a risk factor for the vascular permeability and neovascularization that threatens vision in diabetic retinopathy. Excess reactive oxygen species derived from the Nox (NADPH oxidase) isoforms, Nox1 and Nox4, contributes to vasculopathy in diabetic retinopathy; however, if Nox1/4 inhibition is beneficial in hypertensive diabetic retinopathy is unknown. Here, we determined that diabetic spontaneously hypertensive rats had exacerbated retinal vascular permeability and expression of angiogenic and inflammatory factors, compared with normotensive diabetic Wistar Kyoto rats. GKT136901, a specific dual inhibitor of Nox1 and Nox4, prevented these events in diabetic Wistar Kyoto rats and spontaneously hypertensive rats. Retinal neovascularization does not develop in diabetic rodents, and therefore, the oxygen-induced retinopathy model is used to evaluate this pathology. We previously demonstrated that Nox1/4 inhibition reduced retinal neovascularization in oxygen-induced retinopathy. However, although Nox5 is expressed in human retina, its contribution to retinopathy has not been studied in vivo, largely due to its absence from the rodent genome. We generated transgenic mice with inducible human Nox5 expressed in endothelial cells (vascular endothelial-cadherin+Nox5+ mice). In vascular endothelial-cadherin+Nox5+ mice with oxygen-induced retinopathy, retinal vascular permeability and neovascularization, as well as the expression of angiogenic and inflammatory factors, were increased compared with wild-type littermates. In bovine retinal endothelial cells, which express Nox1, Nox4, and Nox5, Nox1/4 inhibition, as well as Nox5 silencing RNA, reduced the high glucose-induced upregulation of oxidative stress, angiogenic, and inflammatory factors. Collectively, these data indicate the potential of Nox1, Nox4, and Nox5 inhibition to reduce vision-threatening damage to the retinal vasculature.

Comparison of 3 intraosseous catheter sites and methods of determining placement success in cadaver rabbits.

The study goals were to determine if intraosseous (IO) catheters can be placed with greater success into the humerus, femur, or tibia of cadaver rabbits, and to evaluate the accuracy of perceived success (PS) and objective clinical success (OCS) criteria against true intramedullary catheterization confirmed by fluoroscopy. This was a prospective study utilizing 12 rabbit cadavers. Twenty-two participants attempted IO catheter placement at 3 sites. Perceived success, OCS, and fluoroscopic true success (FTS) were recorded. A Fisher's exact test was used to compare PS, OCS, and FTS, and FTS rates between sites (P < 0.05). A Wilcoxon test was used to compare speed of placement (P < 0.05). Overall, of 66 attempts, PS was reported in 86.4%, OCS was documented in 62.1%, FTS was confirmed in 43.9%. Perceived success and OCS overestimated FTS (P ≤ 0.01 and P = 0.027, respectively). Confirmation of FTS occurred in 10/22 (45.5%) humeral, 5/22 (22.7%) femoral, and 14/22 (63.6%) tibial (P = 0.03) attempts. Median time until placement for the humerus was 37.5 seconds (range: 15 to 125 seconds); the femur 135 seconds (range: 91 to 148 seconds); the tibia 49 seconds (range: 19 to 150 seconds). The humerus and tibia were faster to catheterize than the femur (P = 0.01 and 0.03, respectively). Participant PS and OCS criteria overestimated FTS. The humerus or tibia may be more successful and are faster to catheterize.

Les objectifs de la présente étude étaient de déterminer si des cathéters intra-osseux (IO) peuvent être placés avec plus de succès dans l'humérus, le fémur ou le tibia de cadavres de lapins, et d'évaluer la précision des critères du succès perçu (PS) et du succès clinique objectif (OCS) versus le cathétérisme intramédullaire réel confirmé par fluoroscopie. Il s'agissait d'une étude prospective utilisant 12 cadavres de lapin. Vingt-deux participants ont tenté le placement des cathéters IO aux trois sites. Le PS, l'OCS et le succès réel par fluoroscopie (FTS) furent notés. Un test exact de Fisher fut utilisé pour comparer PS, OCS, et FTS, et les taux de FTS entre les sites (P < 0,05). Un test de Wilcoxon a été utilisé pour comparer la vitesse de placement (P < 0,05). Globalement, des 66 essais, PS a été rapporté dans 86,4 % des cas, OCS a été documenté dans 62,1 % des cas, et FTS a été confirmé dans 43,9 % des cas. Le PS et l'OCS surestimaient le FTS (P ≤ 0,01 et P = 0,027, respectivement). La confirmation de FTS s'est produite dans 10/22 (45,5 %) des essais sur l'humérus, 5/22 (22,7 %) des essais sur le fémur, et 14/22 (63,6 %) des essais sur le tibia (P = 0,03).Le temps médian du placement pour l'humérus était de 37,5 secondes (écart : 15 à 125 secondes); pour le fémur de 135 secondes (écart : 91 à 148 secondes); et pour le tibia de 49 secondes (écart : 19 à 150 secondes). Le cathétérisme de l'humérus et du tibia étaient plus rapides que celui du fémur (P = 0,01 et 0,03, respectivement). Les critères pour le PS et l'OCS des participants surestimaient le FTS. L'humérus et le tibia sont plus rapides à cathétériser et le taux de succès est meilleur.(Traduit par Docteur Serge Messier).

PBI-4050 via GPR40 activation improves adenine-induced kidney injury in mice.

PBI-4050 (3-pentylbenzenacetic acid sodium salt), a novel first-in-class orally active compound that has completed clinical Phases Ib and II in subjects with chronic kidney disease (CKD) and metabolic syndrome respectively, exerts antifibrotic effects in several organs via a novel mechanism of action, partly through activation of the G protein receptor 40 (GPR40) receptor. Here we evaluate the effects of PBI-4050 in both WT and Gpr40-/- mice on adenine-induced tubulointerstitial injury, anemia and activation of the unfolded protein response (UPR) pathway. Adenine-induced CKD was achieved in 8-week-old C57BL/6 mice fed a diet supplemented with 0.25% adenine. After 1 week, PBI-4050 or vehicle was administered daily by oral-gavage for 3 weeks. Gpr40-/- mice were also subjected to adenine-feeding, with or without PBI-4050 treatment. PBI-4050 improved renal function and urine concentrating ability. Anemia was present in adenine-fed mice, while PBI-4050 blunted these effects and led to significantly higher plasma erythropoietin (EPO) levels. Adenine-induced renal fibrosis, endoplasmic reticulum (ER) stress and apoptosis were significantly decreased by PBI-4050. In parallel, Gpr40-/- mice were more susceptible to adenine-induced fibrosis, renal function impairment, anemia and ER stress compared with WT mice. Importantly, PBI-4050 treatment in Gpr40-/- mice failed to reduce renal injury in this model. Taken together, PBI-4050 prevented adenine-induced renal injury while these beneficial effects were lost upon Gpr40 deletion. These data reinforce PBI-4050's use as a renoprotective therapy and identify GPR40 as a crucial mediator of its beneficial effects.

Podocyte NADPH Oxidase 5 Promotes Renal Inflammation Regulated by the Toll-Like Receptor Pathway.

AIMS: Oxidative stress associated with a proinflammatory state occurs in endothelial dysfunction, hypertension, chronic kidney disease, and diabetes. The NADPH oxidase (Nox) family of reactive oxygen species (ROS) generating enzymes is implicated in these processes, yet little information regarding the role of Nox5 is available. Our aim was to investigate the role of Nox5 in promoting renal inflammation and identify mechanisms regulating its activity. RESULTS: Mice with podocyte-specific Nox5 (Nox5pod+) expression demonstrated greater glomerular inflammation and increased expression of Toll-like receptors (TLRs) and proinflammatory cytokines. In a lipopolysaccharide (LPS) model of acute kidney injury, Nox5pod+ and control littermates exhibited increased TLR and Nox1 expression. Compared with control littermates, Nox5pod+ animals developed greater glomerular inflammation and ROS production. Immortalized human podocytes (hPODs) incubated with LPS demonstrated TLR induction, increased Nox5 expression, and enhanced ROS production. Inhibition of interleukin-1 receptor-associated kinases (IRAK)-1 and -4 that lie downstream of TLR inhibited LPS-induced ROS production. Interaction between IRAK1 and Nox5 was confirmed by coimmunoprecipitation. Furthermore, LPS treatment of hPODs resulted in phosphorylation of threonine residue(s) in Nox5 that was attenuated by an IRAK1/4 inhibitor. Innovation and Conclusion: These results are the first to demonstrate that Nox5 is a downstream target of the TLR pathway and that Nox5-derived ROS may be modulated by IRAK1/4 activity. Nox5-derived ROS in podocytes can promote a proinflammatory state in the kidney via induction of cytokine expression and upregulation of TLRs leading to a feed-forward loop in which TLR activation enhances Nox5-mediated ROS production.

GRK2 knockdown in mice exacerbates kidney injury and alters renal mechanisms of blood pressure regulation.

The renin-angiotensin system regulates blood pressure and fluid balance in the body primarily via angiotensin receptor 1 (AT1R). Renal AT1R was found to be primarily responsible for Ang II-mediated hypertension. G protein-coupled receptor kinase 2 (GRK2) modulates AT1R desensitization and increased GRK2 protein expression is reported in hypertensive patients. However, the consequences of GRK2 inhibition on kidney functions remain unknown. We employed shGRK2 knockdown mice (shGRK2 mice) to test the role of GRK2 in kidney development and function that can be ultimately linked to the hypertensive phenotype detected in shGRK2 mice. GRK2 knockdown reduced kidney size, nephrogenesis and glomerular count, and impaired glomerular filtration. Glomerular damage in adult shGRK2 mice was associated with increased renin- and AT1R-mediated production of reactive oxygen species. The AT1R blocker, Losartan, normalized elevated blood pressure and markedly improved glomerular filtration in the shGRK2 knockdown mice. Our findings provide evidence for the crucial role of GRK2 in renal regulation of blood pressure. It also suggests that the detrimental outcomes of GRK2 inhibitors on the kidney should be carefully examined when used as antihypertensive.

A Newly Discovered Antifibrotic Pathway Regulated by Two Fatty Acid Receptors: GPR40 and GPR84.

Numerous clinical conditions can lead to organ fibrosis and functional failure. There is a great need for therapies that could effectively target pathophysiological pathways involved in fibrosis. GPR40 and GPR84 are G protein-coupled receptors with free fatty acid ligands and are associated with metabolic and inflammatory disorders. Although GPR40 and GPR84 are involved in diverse physiological processes, no evidence has demonstrated the relevance of GPR40 and GPR84 in fibrosis pathways. Using PBI-4050 (3-pentylbenzeneacetic acid sodium salt), a synthetic analog of a medium-chain fatty acid that displays agonist and antagonist ligand affinity toward GPR40 and GPR84, respectively, we uncovered an antifibrotic pathway involving these receptors. In experiments using Gpr40- and Gpr84-knockout mice in models of kidney fibrosis (unilateral ureteral obstruction, long-term post-acute ischemic injury, and adenine-induced chronic kidney disease), we found that GPR40 is protective and GPR84 is deleterious in these diseases. Moreover, through binding to GPR40 and GPR84, PBI-4050 significantly attenuated fibrosis in many injury contexts, as evidenced by the antifibrotic activity observed in kidney, liver, heart, lung, pancreas, and skin fibrosis models. Therefore, GPR40 and GPR84 may represent promising molecular targets in fibrosis pathways. We conclude that PBI-4050 is a first-in-class compound that may be effective for managing inflammatory and fibrosis-related diseases.