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Introduction. This study addressed the hypothesis that subtotal nephrectomy associated with a high-phosphorus diet (5/6Nx + P) in rats represents a suitable animal model to mimic the cardiovascular consequences of chronic kidney disease (CKD) including calcified aortic valve disease (CAVD). Indeed, the latter contributes to the high morbidity and mortality of CKD patients and sorely lacks preclinical models for pathophysiological and pharmacological studies. Methods. Renal and cardiovascular function and structure were compared between sham-operated and 5/6 Nx rats + P 10 to 12 weeks after surgery. Results. As expected, 11 weeks after surgery, 5/6Nx + P rats developed CKD as demonstrated by their increase in plasma creatinine and urea nitrogen and decrease in glomerular filtration rate, estimated by using fluorescein-isothiocyanate-labelled sinistrin, anemia, polyuria, and polydipsia compared to sham-operated animals on a normal-phosphorus diet. At the vascular level, 5/6Nx + P rats had an increase in the calcium content of the aorta; a decrease in mesenteric artery dilatation in response to a stepwise increase in flow, illustrating the vascular dysfunction; and an increase in blood pressure. Moreover, immunohistology showed a marked deposition of hydroxyapatite crystals in the aortic valve of 5/6Nx + P rats. Echocardiography demonstrated that this was associated with a decrease in aortic valve cusp separation and an increase in aortic valve mean pressure gradient and in peak aortic valve velocity. Left-ventricular diastolic and systolic dysfunction as well as fibrosis were also present in 5/6Nx + P rats. Conclusion. This study demonstrates that 5/6Nx + P recapitulates the cardiovascular consequences observed in humans with CKD. In particular, the initiation of CAVD was shown, highlighting the interest of this animal model to study the mechanisms involved in the development of aortic stenosis and test new therapeutic strategies at an early stage of the disease.

OBJECTIVE Angiopoietins (Angs) regulate endothelial permeability. Ang-1 and 2 (Ang-1 and Ang-2) are implied in endothelial stability through an antagonism effect. The objectives of the present study were to describe and compare changes in Ang levels after transcatheter aortic valve replacement (TAVR) and surgical aortic valve replacement (SAVR). DESIGN A prospective, single-center study. PARTICIPANTS Adult patients with aortic stenosis scheduled for SAVR or TAVR. INTERVENTIONS None. MEASUREMENTS AND MAIN RESULTS Ang-1 and Ang-2 were measured using an enzyme-linked immunosorbent assay right before surgery (T0), at the end of surgery (T1), and at day one (T2). Sixty consecutive patients (SAVR group [n = 30] and TAVR group [n = 30]) were included between January and June 2017. Ang-1 decreased significantly after both TAVR (T0: 3,663 [2,602-4,262]; T1: 1,611 [981-2,409]; T2: 1,082 [652-1,589] ng/mL; p < 0.0001) and SAVR (T0: 1,603 [975-2,849]; T1: 783 [547-1,024]; T2: 828 [460-1,227] ng/mL; p = 0.0001). Ang-2 increased significantly after SAVR (T0: 2,472 [1,502-3,622]; T1: 2,997 [1,759-3,839]; T2: 5,421 [3,557-7,087] ng/mL; p < 0.0001) but did not change markedly after TAVR (T0: 3,343 [2,661-6,272]; T1: 3,788 [2,574-5,016]; T2: 3,446 [3,029-6,313] ng/mL; p = 0.066). Among patients with paravalvular leakage, the changes in the plasma Ang-2 level and the Ang-2/Ang-1 ratio were greater. CONCLUSION SAVR induces greater alterations of Ang homeostasis than TAVR, confirming a role for the use of cardiopulmonary bypass. Paravalvular leakage after TAVR is associated with Ang changes similar to those observed with SAVR.

Background The uraemic toxins that accumulate as renal function deteriorates can potentially affect drug pharmacokinetics. This study’s objective was to determine whether plasma concentrations of certain uraemic toxins are correlated with blood concentrations of two immunosuppressants. Methods DRUGTOX was a cross-sectional study of 403 adult patients followed up after kidney transplantation and who had undergone therapeutic drug monitoring (TDM) of calcineurin inhibitors (tacrolimus or cyclosporin) between August 2019 and March 2020. For each patient, immunosuppressant trough concentrations (C0) were measured in whole blood samples and then normalized against the total daily dose (C0:D ratio). The sample was assayed for five uraemic toxins [urea, trimethylamine N-oxide (TMAO), indole acetic acid (IAA), p-cresylsulphate (PCS) and indoxylsulphate (IxS)] using liquid chromatography–tandem mass spectrometry. Results The median age was 56 years [interquartile range (IQR) 48–66] and the median estimated glomerular filtration rate was 41 mL/min/1.73 m2 (IQR 30–57). Age, sex, body mass index (BMI), urea, IxS and PCS were significantly associated with an increment in the tacrolimus C0:D ratio. A multivariate analysis revealed an independent association with IxS [odds ratio 1.36 (95% confidence interval 1.00–1.85)] after adjustment for sex, age and BMI, whereas adjustment for age weakened the association for PCS and urea. In a univariate logistic analysis, age, sex, BMI and the TMAO level (but not PCS, IxS, IAA or urea) were significantly associated with an increment in the cyclosporine C0:D ratio. Conclusions Even though TDM and dose adaptation of immunosuppressants keep levels within the therapeutic window, increased exposure to tacrolimus (but not cyclosporine) is associated with an accumulation of PCS, IxS and urea.

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Pea (Pisum sativum L.) is subjected to attack by certain soil-borne phylogenetic of fungus-like eukaryotic microorganisms such as Pythium debaryanum and soil-borne fungi Rhizoctonia solani and Fusarium solani which cause damping-off diseases. Isolation of associated fungi and P. debaryanum and three species of Trichoderma was carried out from soil rhizosphere of pea plants. Antagonistic effect of Rhizobium leguminosarum combined with Trichoderma lignorum, T. longibrachiatum and T. koningii against pathogenic fungi was investigated in vitro and in vivo under greenhouse conditions. The effect of combinations between the tested Rhizobium sp. and the other tested Trichoderma spp. on the disease incidence caused by the pathogenic microorganisms was evaluated when used as seed and soil treatments. Disease assessments, nitrogen fixation and yield parameters after 50 and 90 days from sowing in comparison with un-treated and fungicide treatments were recorded. Mycoparasitic activity of the tested Trichoderma spp. against each of the pathogenic fungi and Pythium debaryanum was studied using scanning electron microscope. Results showed that the soil treatments were more effective in controlling damping-off disease than seed treatments in overall experiments. Combination of R. leguminosarum with T. longibrachiatum gave the best results in reducing percentage of post-emergence damping-off (13.33 and 6.67%) and root rot (14.72 and 9.58%) caused by P. debaryanum and R. solani, respectively. Survived plants, nitrogen fixation and yield parameters were also increased. Treatment of R. leguminosarum combined with T. koningii against infection by F. solani gave the best results in reducing percentages of post-emergence damping-off (6.67%) and root rot (13.06%) with increment of survived plants, nitrogen fixation and yield parameters. In conclusion, combinations of R. leguminosarum with the Trichoderma species were effective than application of each one alone against pea damping-off disease.

BACKGROUND AND AIMS Calcific aortic valve disease (CAVD) results from fibro-calcific degeneration of the aortic valve leaflets, causing haemodynamic disorders and major cardiovascular complications. In chronic kidney disease (CKD) patients, this pathological remodelling is more frequent, appears earlier, progresses more rapidly and leads to a mortality rate three times higher than in the general population. The uremic toxin indoxyl-sulfate (IS) is a powerful predictor of cardiovascular mortality in CKD patients and a strong promoter of aortic calcification. However, its role in aortic valve leaflets mineralization has never been studied. This work aimed to evaluate whether IS can influence the mineralization of primary human valvular interstitial cells (hVICs) cultured in vitro. METHOD Primary hVICs were isolated from aortic tricuspid valves collected from patients with CAVD undergoing valve replacement surgery at the Amiens Picardie University Hospital. The concentrations of IS used in this study were selected according to the EUTOX registry: IS normal [Isn]: 0.5 µg/mL, IS uremic [Isu]: 37 µg/mL and IS maximum [Ismax]: 233 µg/mL. Human VICs mineralization were induced by exposure to an osteogenic medium containing 2.4 mmol/L calcium and 2.5 mmol/L phosphate (pro-calcific condition) and quantified by the o-cresolphthalein complexone method. The osteogenic transition of hVICs was assessed by qRT-PCR following Bmp2 and Runx2 mRNA expression. The cells’ inflammatory potential was assessed by qRT-PCR and ELISA following IL-1β, IL-6 and TNF-α expression and secretion. Activation of NFkB pathway was studied by analysing the phosphorylation of P65 via western blot. A siRNA approach was used to determine the signaling pathway involved in the effects of IS. RESULTS In pro-calcific conditions, IS increased hVICs osteogenic transition and calcification in a concentration-dependent manner, an effect that was blocked in hVICs transfected with siRNA targeting AhR expression. Data obtained by western blot showed that the exposure to [ISu] and [ISmax] promoted P65 phosphorylation in an AhR dependent manner. Blockade of the NFкB pathway with siRNA targeting P65 inhibited IS-induced expression of Bmp2 and RunX2 as well as hVICs mineralization, suggesting that hVICs inflammation plays a key role in the procalcific effects of IS. Confirming this hypothesis, the exposure to [ISu] and [ISmax] increased the expression of IL-1β, IL-6 and TNF-α transcripts, an effect abolished by silencing AhR and P65. Interestingly, hVICs did not secrete TNF-α and produced only a very low quantity of IL-1β in response to IS. By contrast, the exposure to [ISu] and [ISmax] induced an important release of IL-6 in hVICs culture medium, which was blocked when AhR and P65 were silenced. In this model, the use of an antibody neutralizing IL-6 completely abolished IS-induced hVICs osteogenic transition and calcification. CONCLUSION Our study demonstrates that IS displays direct pro-calcific effects on primary hVICs through AhR-dependent activation of the NFkB pathway and subsequent release of IL-6. We report that the autocrine action of IL-6 is responsible for IS-induced hVICs osteogenic transition and calcification. The observation that antibodies neutralizing IL-6 can block IS pro-calcific effects might be of particular interest knowing that interventional studies evaluating the cardiovascular impact of anti-IL-6 antibodies in patients with CKD are underway.

CXCR1 and CXCR2 chemokine receptors, mainly activated by interleukin 8 (IL-8 or CXCL8), are expressed in a variety of cells including, leukocytes, fibroblasts, endothelial cells, and smooth muscle cells. Numerous intracellular mediators are activated by these G protein-coupled receptors based on several factors, including the nature of the ligand, its concentration, and the binding sites with the receptor, levels of the receptor, cell type, and stimulatory environment. Much focus is currently being directed towards CXCR1/2 inhibitors, as these receptors primarily induce the chemotaxis of leukocytes, especially neutrophils, during inflammation, a key process in cardiovascular disease (CVD) progression. CXCR1/2 inhibitors show beneficial effects in various animal models of CVD. These effects include reducing the atherosclerotic plaque area, improving the serum lipid profile, attenuation of the damage following ischemia-reperfusion, the regulation of blood pressure, and the restriction of cardiac remodeling. Based on these encouraging results, testing CXCR1/2 inhibitors in clinical trials could be of a great importance to limit the inflammatory complications associated with CVDs.

In the current study, evaluation the ability of three isolates of Sclerotium rolfsii isolated from tomato, bean and cabbage to infect their host plants and two non-host plants was carried out. Tomato plants were strongly affected with the three isolates of the pathogen followed by cabbage plants, whereas common bean plants were the least in this respect. Six species of Trichoderma were examined for their bio-control potentials against S. rolfsii the causative of root rot in tomato, common bean and cabbage. In vitro antagonistic test showed that T. koningii exhibited high inhibitory effect as the percentage of inhibition value was higher in case of S. rolfsii (tomato isolate) followed by T. harzianum with S. rolfsii (bean isolate) and S. rolfsii (cabbage isolate). Under greenhouse conditions; T. koningii, T. viride and T. harzianum showed the highest antagonistic effect against the three isolates of S. rolfsii in pots experiment. Microscopic examinations showed that most of Trichoderma spp. grew over mycelia of the tested pathogen with surrounded, coiling, lysis of hyphae and collapse of mycelium. T. koningii and T. viride exhibited the best performance regarding to the defense enzyme secretion; chitinase and β-glucanase followed by T. harzianum. Also, all the tested Trichoderma spp. produced Indole acetic acid (IAA) and Gibberellic acid (GA) as a plant growth promoting substances in variable values.

Resume Au niveau de la paroi vasculaire, on peut distinguer de la peripherie du vaisseau vers la lumiere, l’adventice, la media composee essentiellement de cellules musculaires lisses et l’intima composee essentiellement de cellules endotheliales. La vasomotricite est la capacite du vaisseau a se contracter et a se relâcher. Pour cela, l’endothelium repond a des stimuli et les traduit en reponses biologiques qui seront transmises au muscle lisse. Le muscle lisse possede egalement l’ensemble des systemes lui permettant de se contracter ou de se relâcher independamment des signaux produits par l’endothelium. On distingue donc une vasomotricite liee a l’action directe du muscle lisse et une vasomotricite dependante de l’endothelium. Les reponses vasomotrices du muscle lisse sont liees a l’activation de voies de signalisation entrainant des variations de concentrations de calcium intracellulaire. L’augmentation de calcium resultant entraine une vasoconstriction alors que la diminution de calcium entraine une vasorelaxation. La vasoconstriction dependante de l’endothelium est liee a la production de facteurs produits par l’endothelium (les derives du metabolisme de l’acide arachidonique : les endoperoxydes, le thromboxane A2, la prostaglandine H2, l’endotheline, les ROS (Reactive Oxygen Species, especes reactives de l'oxygene) et l’angiotensine II) entrainant une contraction du muscle lisse. Les cellules endotheliales sont capables de moduler la vasomotricite grâce a la production de facteurs vasodilatateurs que sont la prostacycline (PG12), le monoxyde d'azote (NO) et le facteur hyperpolarisant derive de l'endothelium (EDHF, endothelium derived hyperpolarizing factor). La fonction vasculaire et son maintien sont un element essentiel garantissant la sante du vaisseau.

Resume Les angiopoietines 1 et 2 (Ang-1 et Ang-2) et leurs recepteurs associes Tie1 et Tie2 representent un systeme de regulation extremement fin de l’endothelium vasculaire. Ang-1 est un facteur secrete dans le microenvironnement endothelial par les pericytes, et agit comme un agoniste du recepteur Tie2 exercant de multiples effets vasculaires protecteurs. A l’inverse, Ang-2, secrete principalement par les cellules endotheliales, bloque la signalisation induite par le recepteur Tie2 et antagonise les effets d’Ang-1, ce qui a pour consequence de destabiliser l’endothelium vasculaire. Le ratio Ang-2/Ang-1 est donc un element cle dans la regulation de la fonction endotheliale. Toutes situations physiopathologiques capables de moduler ce ratio en augmentant la production d’Ang-2 ou en diminuant celle d’Ang-1 pourraient donc etre a l’origine d’une dysfonction endotheliale. C’est ce que l’on observe au cours des etats inflammatoires qu’ils soient aigus, comme au cours du sepsis, ou chroniques, comme au cours des maladies cardio-vasculaires. La mesure des concentrations circulantes d’Ang-1 et d’Ang-2 constitue donc potentiellement un bon marqueur biologique de la dysfonction endotheliale. Les principaux objectifs de cette revue sont, d’une part, de presenter succinctement la structure et les differentes activites biologiques au niveau vasculaire des angiopoietines et, d’autre part, de presenter les differentes etudes cliniques et biologiques dans lesquelles les concentrations circulantes des angiopoietines se sont averees etre de bons marqueurs biologiques de la dysfonction endotheliale.

The role of raw cow's milk and whey (40 and 50% dilution in water) and number of sprays against cucumber powdery mildew caused by Sphaerotheca fuliginea was studied in greenhouse experiments during 2017/18 and 2018/19. As a result of number of sprays, the disease severity percentage and area under the disease progress curve (AUDPC) were affected by the number of sprays with raw milk and whey. The number of sprays played an important role in controlling powdery mildew, and significant differences have occurred among all applied sprays. Four sprays with intervals of one week was the effective treatment in reducing disease severity, AUDPC values, compared to other treatments, i.e., two and three sprays in comparison to the fungicide as well as a control treatment. Additionally, treated plants showed a significant increase in plant growth and yield parameters in case of spraying four times. Also, the activity of defense-related enzymes (catalase, peroxidase, and polyphenoloxidase) and amino acid proline was high in response to increasing the number of sprays at four times compared with untreated plants. Light and scanning electron microscopic observations showed a clear change in the morphology of S. fuliginea spores, in addition to hyphae degradation and loss the vitality of conidia as well as rupturing the content of conidia. The results obtained also indicated that cow's milk and whey dilution effectively reduced the powdery mildew signs and symptoms on infected cucumber plants at 50% with the four sprays treatment being the most effective treatment.

Background: Vascular calcification (VC) is a cardiovascular complication associated with a high mortality rate among patients with diseases such as atherosclerosis and chronic kidney disease. During VC, vascular smooth muscle cells (VSMCs) undergo an osteogenic switch and secrete a heterogeneous population of extracellular vesicles (EVs). Recent studies have shown involvement of EVs in the inflammation and oxidative stress observed in VC. We aimed to decipher the role and mechanism of action of macrophage-derived EVs in the propagation of inflammation and oxidative stress on VSMCs during VC.Methods: The macrophage murine cell line RAW 264.7 treated with lipopolysaccharide (LPS-EK) was used as a cellular model for inflammatory and oxidative stress. EVs secreted by these macrophages were collected by ultracentrifugation and characterized by transmission electron microscopy, cryo-electron microscopy, nanoparticle tracking analysis, and the analysis of acetylcholinesterase activity, as well as that of CD9 and CD81 protein expression by western blotting. These EVs were added to a murine VSMC cell line (MOVAS-1) under calcifying conditions (4 mM Pi—7 or 14 days) and calcification assessed by the o-cresolphthalein calcium assay. EV protein content was analyzed in a proteomic study and EV cytokine content assessed using an MSD multiplex immunoassay.Results: LPS-EK significantly decreased macrophage EV biogenesis. A 24-h treatment of VSMCs with these EVs induced both inflammatory and oxidative responses. LPS-EK-treated macrophage-derived EVs were enriched for pro-inflammatory cytokines and CAD, PAI-1, and Saa3 proteins, three molecules involved in inflammation, oxidative stress, and VC. Under calcifying conditions, these EVs significantly increase the calcification of VSMCs by increasing osteogenic markers and decreasing contractile marker expression.Conclusion: Our results show that EVs derived from LPS-EK–treated-macrophages are able to induce pro-inflammatory and pro-oxidative responses in surrounding cells, such as VSMCs, thus aggravating the VC process.

Like all proteins, plasma albumin can undergo glycation to produce glycated albumin. This glycation will change the structure and therefore functionalities of albumin. Because of its accessibility, its high concentration and its half-life the glycation of albumin is greater than that of haemoglobin. Laboratory measurement of glycated albumin is possible. The value of glycated albumin reflects the glycaemic balance over a period of 3 weeks, and shows its interest in cases where the determination of HbA1c is impossible or deficient. It also seems that the glycated albumin has a prognostic interest in the chronic kidney disease patients dialyzed or not. In some patients, the assay of glycated albumin could replace measurement of fructosamines and provide additional information to HbA1c values.

Ischemic stroke is highly prevalent in chronic kidney disease (CKD) patients and has been associated with a higher risk of neurological deterioration and in-hospital mortality. To date, little is known about the processes by which CKD worsens ischemic stroke. This work aimed to investigate the cellular and molecular mechanism associated with ischemic stroke severity in an in vivo model of CKD. CKD was induced through right kidney cortical electrocautery in 8-week-old female C57BL/6 J mice followed by left total nephrectomy. Transient middle cerebral artery occlusion (tMCAO) was performed 6 weeks after left nephrectomy. Twenty-four hours after tMCAO, the infarct volumes were significantly wider in CKD than in SHAM mice. CKD mice displayed decreased neuroscore, impaired ability to remain on rotarod device, weaker muscular strength and decreased prehensile score. Apoptosis, neuronal loss, glial cells recruitment and microglia/macrophages M1 signature genes CD32, CD86, IL-1β, IL-6, MCP1 and iNOS were significantly increased within ischemic lesions of CKD mice. This effect was associated with decreased AMP kinase phosphorylation and increased activation of the NFΚB pathway. Pharmacological targeting of AMP kinase activity, which is known to block microglia/macrophages M1 polarization, appears promising to improve stroke recovery in CKD.

Introduction and aims Calcific aortic valve disease (CAVD) is the most common heart valve disease in western countries. It has been reported that activation of the calcium-sensing receptor(CaSR) expressed by vascular smooth muscle cells prevents vascular calcification. However, to date, the CaSR's expression and function in cardiac valves have not been studied. The present study sought to evaluate the presence of the CaSR within human valvular interstitial cells (hVICs), assess the CaSR's functionality, and ascertain its involvement in hVIC calcification. Methods and results Data from Western blot, flow cytometry and immunocytochemistry experiments demonstrated that primary hVICs express the CaSR. The receptor was functional, since the incubation of hVICs with the calcimimetic R-568 significantly increased Ca2+-induced ERK1/2 phosphorylation, and exposure to the calcilytic NPS2143 reduced ERK1/2 activation. A reduction in endogenous CaSR expression by hVICs (using siRNA) was associated with significantly lower levels of Ca2+-induced mineralization (quantified using Alizarin Red staining). Similar data were obtained after the pharmacological inhibition of CaSR activity by the calcilytic NPS2143. In contrast, overexpression of a functional CaSR amplified Ca2+-induced calcification. Pharmacological activation of the CaSR with the calcimimetic R-568 showed similar effects. CaSR's procalcific properties are associated with increased osteogenic transition (as characterized by elevated mRNA expression of bone morphogenetic protein 2 and osterix), and reduced the expression of the calcification inhibitor osteopontin. Histological analysis of 12 human aortic tricuspid valves showed that CaSR expression was greater in calcified areas than in non-calcified areas. These data were confirmed by Western blots. Conclusions To the best of our knowledge, this study is the first to have demonstrated that hVICs express a functional CaSR. Taken as a whole, our data suggest that activation of the CaSR expressed by hVICs might be a key promoter of CAVD progression.

Late blight caused by Phytophthora infestans is one of the most devastating diseases on potato worldwide. Partial resistance (PR) to the oomycete pathogen Phytophthora infestans was studied for ten potato cultivars in Beheira, Gharbia and Kafr El-Sheikh governorates in Egypt. Final disease severity (FDS%), area under disease progress curve (AUDPC) and rate of disease increase (r-value) were used as epiphytotic parameters to estimate PR during the growing seasons of 2016 and 2017. The cvs. Burren, Cara and Bellini exhibited the highest levels of PR to late blight, where they showed lowest values of FDS (%), AUDPC and r-value in both seasons of evaluation. Four measures i.e., means x, regression coefficient (b1), standard deviation (S2d) and ecovalance (wi) were used for determining the partial stability resistance of the tested potato cvs., to late blight infection. Stability parameters clearly showed that cv. Bellini was the most stable to late blight under wide range of environmental conditions. The stability analysis can help breeders to monitor the level of resistance of the tested plant material as well as to monitor the disease pressure in different environments.

Background and Aims In addition to affecting glomerular filtration rate, chronic kidney disease (CKD) changes the absorption, distribution and nonrenal clearance of a number of drugs. The uremic toxins that accumulate with as renal function deteriorates can potentially affect drug pharmacokinetics. Given the requirement for therapeutic drug monitoring (TDM) with most immunosuppressants, the latter are good models for understanding the potential impact of uremic toxins on blood concentrations of drugs. We focused on two calcineurin inhibitors (tacrolimus and cyclosporine) with similar pharmacological properties: low but variable oral bioavailability, strong binding to plasma proteins (albumin for tacrolimus and lipoproteins for cyclosporine), metabolism by cytochrome P450 3A4/5, and excretion in the bile. Our starting hypothesis was that uremic toxins influence the exposure of immunosuppressants. The study’s objective was to determine whether plasma concentrations of certain uremic toxins were correlated with blood concentrations of the two immunosuppressants. Method DRUGTOX was a cross-sectional study of 403 adult patients followed up after kidney transplantation at Amiens University Medical Center (Amiens, France) and who had undergone TDM of calcineurin inhibitors between August 4th, 2019, and March 11th, 2020. The patients had been treated daily with oral tacrolimus or cyclosporine. Two groups were constituted: 203 patients with low uremic toxin levels (estimated glomerular filtration rate (eGFR) according to the Modification of Diet in Renal Disease equation >40 mL/min/1.73m2) and 200 patients with high uremic toxin levels (eGFR Results The median [interquartile range] age was 56 [48-66] and the median eGFR was 41 [30-57] ml/min/1.73 m2. The majority of the patients (56.5%) were at CKD stage 3. A total of 248 (61.5%) patients were being treated with tacrolimus and 155 (38.5%) were being treated with cyclosporine. The median time between transplantation and study inclusion was 6.5 years. Plasma urea, TMAO, IAA, PCS and IxS levels rose progressively with the CKD stage. In a univariate logistic analysis, age, sex, BMI, urea, IxS and PCS were significantly associated with an increment in the tacrolimus C0/D ratio. A multivariate analysis revealed an independent association with IxS (odds ratio [95% confidence interval]: 1.36 [1.00; 1.85], after adjustment for sex, age and BMI, whereas adjustment for age weakened the association for PCS (OR: 1.22 (0.99; 1.55)) and urea (OR: 1.71 (0.93; 3.16)). In a univariate logistic analysis, age, sex, BMI, and the TMAO level (but neither PCS, IxS, IAA nor urea) were significantly associated with an increment in the cyclosporine C0/D ratio. Conclusion Even though TDM and dose adaptation of immunosuppressants keeps levels within the therapeutic window, elevated exposure to tacrolimus (but not cyclosporine) is associated with an accumulation of PCS, IxS and urea. This might be explained by an interaction between tacrolimus and highly albumin-bound uremic toxins. Our findings may have implications for other drugs that bind with high affinity to albumin (since an elevated free fraction might lead to unexpected exposures, which can alter their efficacy and/or tolerance).

Background Bone remodeling is a complex process, and many conditions (including drug exposure) lead to osteoporosis. Here, we sought to detect new disproportionality signals for drugs associated with osteoporosis. Methods We performed a disproportionality analysis of the World Health Organization’s VigiBase® pharmacovigilance database through April 12, 2020. The frequency of reports on osteoporosis for all identified drug classes was compared with that for all other drugs and quoted as the reporting odds ratio (ROR) [95% confidence interval (CI)]. Results Of the 7,594,968 cases spontaneously recorded to VigiBase®, 4,758 concerned osteoporosis. New disproportionality signals with a pharmacologically plausible mechanism were found for drugs used in neurology (levodopa (ROR [95%CI]: 10.18 [4.33–25.10]), selective serotonin agonists (4.22 [2.34–7.00]) and memantine (4.10 [1.56–8.93])), hematology (romiplostim (4.93 [1.15–21.10])), pulmonology (macitentan (3.02 [1.84–4.90])), ophthalmology (ranibizumab (3.31 [1.00–10.51])) and rheumatology (tofacitinib (3.65 [3.00–4.40])). The robustness of these new results is supported by the significant RORs for the vast majority of drugs already known to induce osteoporosis and/or increase the fracture risk, namely glucocorticoids, gonadotropin-releasing hormone analogs, anti-aromatases, androgen receptor blockers, thyroid hormones, proton pump inhibitors, thiazolidinediones, vitamin K antagonists, loop diuretics, protease inhibitors, nucleoside and nucleotide reverse transcriptase inhibitors, and enzyme-inducing antiepileptics including barbiturates and derivatives, hydantoin derivatives, carboxamide derivatives and fatty acid derivatives. Conclusion We established up a comprehensive list of drugs potentially associated with osteoporosis and highlighted those with pharmacologically plausible mechanisms leading to bone fragility. Our results might pave the way for additional exploration of these mechanisms.

International audience; Chronic kidney disease (CKD) is a highly prevalent condition and is associated with a high comorbidity burden, polymedication, and a high mortality rate. A number of conventional and nonconventional risk factors for comorbidities and mortality in CKD have been identified. Among the nonconventional risk factors, uremic toxins are valuable therapeutic targets. The fact that some uremic toxins are gut-derived suggests that intestinal chelators might have a therapeutic effect. The phosphate binders used to prevent hyperphosphatemia in hemodialysis patients act by complexing inorganic phosphate in the gastrointestinal tract but might conceivably have a nonspecific action on gut-derived uremic toxins. Since phosphorous is a major nutrient for the survival and reproduction of bacteria, changes in its intestinal concentration may impact the gut microbiota's activity and composition. Furthermore, AST-120 is an orally administered activated charcoal adsorbent that is widely used in Asian countries to specifically decrease uremic toxin levels. In this narrative review, we examine the latest data on the use of oral nonspecific and specific intestinal chelators to reduce levels of gut-derived uremic toxins.

This study addressed the hypothesis that soluble epoxide hydrolase (sEH), which metabolizes endothelium-derived epoxyeicosatrienoic acids, plays a role in vascular calcification. The sEH inhibitor trans-4-(4-(3-adamantan-1-yl-ureido)-cyclohexyloxy)-benzoic acid (t-AUCB) potentiated the increase in calcium deposition of rat aortic rings cultured in high-phosphate conditions. This was associated with increased tissue-nonspecific alkaline phosphatase activity and mRNA expression level of the osteochondrogenic marker Runx2. The procalcifying effect of t-AUCB was prevented by mechanical aortic deendothelialization or inhibition of the production and action of epoxyeicosatrienoic acids using the cytochrome P450 inhibitor fluconazole and the antagonist 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE), respectively. Similarly, exogenous epoxyeicosatrienoic acids potentiated the calcification of rat aortic rings through a protein kinase A (PKA)-dependent mechanism and of human aortic vascular smooth muscle cells when sEH was inhibited by t-AUCB. Finally, a global gene expression profiling analysis revealed that the mRNA expression level of sEH was decreased in human carotid calcified plaques compared to adjacent lesion-free sites and was inversely correlated with Runx2 expression. These results show that sEH hydrolase plays a protective role against vascular calcification by reducing the bioavailability of epoxyeicosatrienoic acids.

International audience; Vascular dysfunction is an essential element found in many cardiovascular pathologies and in pathologies that have a cardiovascular impact such as chronic kidney disease (CKD). Alteration of vasomotricity is due to an imbalance between the production of relaxing and contracting factors. In addition to becoming a determining factor in pathophysiological alterations, vascular dysfunction constitutes the first step in the development of atherosclerosis plaques or vascular calcifications. In patients with CKD, alteration of vasomotricity tends to emerge as being a new, less conventional, risk factor. CKD is characterized by the accumulation of uremic toxins (UTs) such as phosphate, para-cresyl sulfate, indoxyl sulfate, and FGF23 and, consequently, the deleterious role of UTs on vascular dysfunction has been explored. This accumulation of UTs is associated with systemic alterations including inflammation, oxidative stress, and the decrease of nitric oxide production. The present review proposes to summarize our current knowledge of the mechanisms by which UTs induce vascular dysfunction.

Background and Aims Ischemic stroke is highly prevalent in patients with chronic kidney disease (CKD) and has been associated with poor outcomes. In experimental studies, CKD mice display wider and more inflammatory ischemic lesions as well as poorer functional outcomes than mice with normal renal function. In rodents with normal renal function, treatment with a mineralocorticoid receptor antagonist decreases brain ischemic lesion and improves functional outcomes. The goal of this study was to investigate the impact of a treatment with spironolactone, a mineralocorticoid receptor antagonist, on stroke outcomes in CKD mice. Method Male C57BL/6J mice were randomly assigned to CKD or SHAM groups. CKD was induced through right kidney cortical electrocautery in 8 week-old mice followed 2 weeks later by left total nephrectomy. Mice received spironolactone (50 µg/kg/day) or placebo (subcutaneous tablets, Innovative Research of America) for 6 weeks until stroke induction by transient middle cerebral artery occlusion (tMCAO). Neurological deficits were monitored daily, 3 days before and 1 day after stroke induction. Mice were euthanized 24 hours after tMCAO. Total infarct volumes and apoptosis were determined by cresyl violet staining and TUNEL respectively. Immunohistochemical detection of neurons was performed using an anti-NeuN antibody. To characterize the pro-inflammatory polarization of microglia/macrophages, M1 signature genes’ expression (CD-32, IL-1β and IL-6) was analyzed by qPCR. Results Untreated CKD mice displayed wider ischemic lesions (40.3 ± 4.5 versus 23.8 ± 3.9 mm3; p < 0.05) and poorer neurological score, grip strength and prehensile abilities compared to untreated sham mice. Pro-inflammatory M1 signature genes’ expression (CD-32, IL-1β and IL-6) was significantly increased in the ischemic hemisphere of CKD mice compared to the non-ischemic hemisphere. Treatment with spironolactone decreased ischemic lesion (19.3 ± 5.1 mm3; p < 0.05) and improved neurological score, grip strength and prehensile abilities in CKD mice. Treatment with spironolactone also reduced apoptosis, neuronal loss and microglia/macrophages pro-inflammatory M1 signature genes’ expression in CKD mice. In contrast, treatment with spironolactone had no effect in sham mice. Conclusion These data demonstrate that the mineralocorticoid receptor could be a promising therapeutic target to decrease brain inflammation and improve stroke recovery in CKD.

International audience; In chronic kidney disease (CKD), calcium-sensing receptor (CaSR) expression and function have been extensively studied in parathyroid tissue and vascular tissues. To examine whether similar changes occurred in other tissues, we measured total and surface CaSR expression in monocytes of patients with various stages of CKD and healthy volunteers respectively in cross-sectional studies. We further explored in vitro the impact of uremic serum on CaSR expression in monocytes (U937 and THP-1 cell lines), and whether human peripheral blood mononuclear cells or U937 and THP-1 monocytes might modify vascular calcium deposition in rat carotid arteries in vitro. CKD was associated with a decrease in peripheral blood mononuclear cell CaSR expression both in total and at the monocyte surface alone (43% and 34%, respectively in CKD stages 4-5). This decrease was associated with a reduction in the ability of monocytes to inhibit vascular calcification in vitro. Pretreatment with the calcimimetic NPSR568 of peripheral blood mononuclear cells isolated from patients with CKD significantly improved monocyte capacity to reduce carotid calcification in vitro. The fewer peripheral blood mononuclear cells expressing cell surface CaSR, the more calcimimetic treatment enhanced the decrease of carotid calcium content. Thus, we demonstrate that monocyte CaSR expression is decreased in patients with CKD and provide in vitro evidence for a potential role of this decrease in the promotion of vascular calcification. Hence, targeting this alteration or following monocyte CaSR expression as an accessible marker might represent a promising therapeutic strategy in CKD-associated arterial calcification.

Strawberry (Fragaria x ananassa Duch.) is a widely grown vegetable crop in Egypt. During a routine survey,wilt and root rot symptoms were observed on strawberry plants cv. Festival. Samples were collected and isolation was done on potato dextrose agar (PDA) medium. The frequently isolated fungi were Pestalotiopsis spp. along with other associated fungi. Koch’s postulates were fulfilled against two strawberry cultivars Fortuna and Festival under greenhouse conditions. Pestalotiopsis fungal isolates were identified by amplification and sequencing of ITS, TEF-1α and β-tubulin gene regions. The BLASTn search of GenBank database revealed that all obtained sequences exhibited similarity of 99–100% to Neopestalotiopsis rosae. Four fungicides were evaluated against N. rosae through in vitro and in vivo trials. In vitro trials revealed that fungicides Thiram and Hymexazole inhibited the mycelial growth of N. rosae at concentrations 1000 and 1250 ppm, respectively. In vivo trials showed that Thiram reduced significantly disease severity of crown and root rot on both cultivars.

In chronic kidney disease (CKD), the progressive decrease in renal function leads to disturbances of mineral metabolism that generally cause secondary hyperparathyroidism. The increase in serum parathyroid hormone is associated with reduced serum calcium and calcitriol levels and/or increased serum fibroblast growth factor-23 and phosphate levels. The resulting CKD-associated disorder of mineral and bone metabolism is associated with various other metabolic dysregulations such as acidosis, malnutrition, inflammation, and accumulation of uremic toxins. It favors the occurrence of vascular calcification, which results from an imbalance between numerous inhibitors and promoters of soft-tissue mineralization. This review provides an overview of the most recent state of knowledge concerning the mechanisms that lead to the development of vascular calcification in the CKD setting. It further proposes directions for potential new therapeutic targets.

The plant pathogenic oomycete, Phytophthora infestans, is the causal agent of late blight disease in tomato and potato. For characterizing Egyptian P. infestans isolates by DNA marker analysis, 40 isolates of P. infestans were collected from different locations in Egypt during two growing seasons (2012/2013 and 2013/2014). The 40 isolates were grouped into seven genotypes, in which 24 alleles were detected. The identified genotypes were not completely associated with geographic location and sample collection years. These results provide genetic and geographical information for developing a program to manage late blight disease.

Cardiovascular calcification is a growing burden and a leading contributor to acute cardiovascular events, but no therapeutics are currently available. Recently, Choi et al. [(1)][1] put forward dipeptidyl peptidase (DPP)-4 as a new pharmacological target to prevent aortic valve calcification. DPP

Fusarium oxysporum f. sp. cucumerinum (FOC) is one of the important fungi that infect cucumber plants, causing significant yield losses. Plant pathogenic fungi differ in their disease severity and aggressiveness degree with diverse infection modes governed by specialized proteins involved in signaling pathways leading to pathogenicity, the area which is less explored. To understand the pathogenicity and virulence variation of FOC isolates, a comprehensive proteomic study was done to compare the polypeptide patterns of mycelial, conidial, and secreted proteins of the highly pathogenic (hp-FOC) and weakly pathogenic (wp-FOC) isolates. Two-dimensional gel electrophoresis (2-DE) coupled with LC-MS/MS analysis led to the identification of shared and unique proteins involved in metabolism and pathogenicity in both fungal isolates. This study may outline the biological and molecular basis of the different fungal structures to understand the mechanism of their action inside the host tissues. The determination of proteins that express the ability of the pathogen to induce pathogenesis may aid in detecting specialized inhibitors that suppress pathogenic protein activities to control the disease, this in turn may contribute to reduce the excessive use of fungicides, which further enhance the productivity, crop quality, and environmental safety.