Rare copy number variations (CNVs) are a major cause of genetic diseases. Simple targeted methods are required for their confirmation and segregation analysis. We developed a simple and universal CNV ...assay based on digital PCR (dPCR) and universal locked nucleic acid (LNA) hydrolysis probes.
We analyzed the mapping of the 90 LNA hydrolysis probes from the Roche Universal ProbeLibrary (UPL). For each CNV, selection of the optimal primers and LNA probe was almost automated; probes were reused across assays and each dPCR assay included the CNV amplicon and a reference amplicon. We assessed the assay performance on 93 small and large CNVs and performed a comparative cost-efficiency analysis.
UPL-LNA probes presented nearly 20000000 occurrences on the human genome and were homogeneously distributed with a mean interval of 156 bp. The assay accurately detected all the 93 CNVs, except one (<200 bp), with coefficient of variation <10%. The assay was more cost-efficient than all the other methods.
The universal dPCR CNV assay is simple, robust, and cost-efficient because it combines a straightforward design allowed by universal probes and end point PCR, the advantages of a relative quantification of the target to the reference within the same reaction, and the high flexibility of the LNA hydrolysis probes. This method should be a useful tool for genomic medicine, which requires simple methods for the interpretation and segregation analysis of genomic variations.
•A geochemical approach was used to investigate wastewater irrigated soil and crop quality.•Specific drip irrigation can lead to localized metals accumulation in the early cycles.•Transient ...significant Na and B contents can develop under the drip emitter.•The main Pb source in the Cretan soil was petrol-Pb aerosols.
To fight against sanitary risks due to the use of raw or insufficiently treated wastewaters, an irrigation system combining a farm-scale decentralized wastewater treatment and an improved drip irrigation management was developed. The whole soil-water-plant system was monitored on an experimental tomato field in Crete to assess the potential element accumulation due to drip irrigation with secondary treated wastewaters during 3 years. Although a decrease of the majority of element average concentrations (Mn, As, Cr, Cu, Pb, Zn and B) was observed in irrigated soils possibly due to crop export, increasing amounts of elements, especially for As, Cd, Zn, Cu, Ni and Na were measured in the bio-accessible and mobilizable fractions of the soil. Moreover, we show that surface drip irrigation can lead to very localized accumulation of trace metals driven by geochemical processes as pointed out by soil mapping. Transient but significant Na and B contents can also develop under the emitter in the soil fractions during secondary treated wastewater irrigation. Pb displayed no spatial relation with the drip emitter and its main sources were petrol-Pb aerosols and Saharan dusts. The trace metal contents and salinity indicators in crops stayed below the regulatory thresholds. Sequential dissolutions combined with the Pb isotopic tool traced element mobility and dynamics in an irrigated soil at short term, and were useful to distinguish between medium-term experiment at bulk soil scale and short-term impacts of secondary treated wastewaters irrigation at the drip emitter scale.
Deferasirox is the first oral iron chelator and, as such, is widely used for the treatment of chronic iron overload. However, recent data from large studies confirmed the renal toxicity of ...deferasirox. We report a case of Fanconi syndrome associated with acute renal failure in a patient receiving deferasirox. In particular, new insights regarding the pathophysiology of the renal disease due to this treatment are discussed. This case highlights the importance of a careful monitoring of kidney function, markers of proximal tubulopathy and ferritinaemia in patients receiving deferasirox.
Salinization in coastal aquifers is a global phenomenon resulting from the overexploitation of scarce water resources. The Gaza Strip is one of the most severe cases of salinization, as accelerated ...degradation of the water quality endangers the present and future water supply for over 1 million people. We investigate the chemical and isotopic (87Sr/86Sr, delta11B, delta18O, delta2H, and delta34S(SO4)) compositions of groundwater from the southern Mediterranean coastal aquifer (Israel) and the Gaza Strip in order to elucidate the origin of salinity and boron contamination. The original salinity in the eastern part of the aquifer is derived from discharge of saline groundwater from the adjacent Avedat aquitard (Na/Cl < 1, 87Sr/86Sr approximately 0.7079, and delta11B approximately 40 per thousand). As the groundwater flows to the central part of the aquifer, a dramatic change in its composition occurs (Na/Cl > 1, high B/Cl, SO4/Cl, and HCO3, 87Sr/86Sr approximately 0.7083; delta11B approximately 48 per thousand), although the delta18O-delta2H slope is identical to that of the Avedat aquitard. The geochemical data suggest that dissolution of pedogenic carbonate and gypsum minerals in the overlying loessial sequence generated the Ca-rich solution that triggered base exchange reactions and produced Na- and B-rich groundwater. The geochemical data show that most of the salinization process in the Gaza Strip is derived from the lateral flow of the Na-rich saline groundwater, superimposed with seawater intrusion and anthropogenic nitrate pollution. The methodology of identification of multiple salinity sources can be used to establish a long-term management plan for the Gaza Strip and can also be implemented to understand complex salinization processes in other similarly stressed coastal aquifers.
The Corsica-Sardinia batholith in the southern realm of the Hercynian belt of Europe shows evidence for gravitational collapse of this part of the mountain belt, causing major felsic and mafic ...magmatism. The latest intrusions are composed of leucomonzogranite and late metaluminous and alkaline granite, associated with tholeiitic layered complexes and dykes. Three dating methods on zircon (Pb-evaporation, ID-TIMS and SHRIMP) were used to unravel the chronology of these felsic and mafic rocks. Dating of zircons by the conventional U–Pb method, using TIMS after zircon dissolution, achieved an analytical uncertainty of 1 Ma for favourable cases. The TIMS Pb-evaporation technique resulted in ages with an uncertainty range of 4 to 8 Ma. After 15 to 20 analyses with the SHRIMP method, a precision ranging from 2 to 5 Ma was obtained (all at 2
σ). The three methods applied to the same zircon population extracted from four A-type granites, show that the uncertainty ranges within 2–5 Ma according to the sample. This error seems to correspond to the real geochronological uncertainty that can be achieved.
The results obtained show that all six tested alkaline granites were emplaced during a very short interval of about 3–5 Ma at about 288 Ma, almost contemporaneous with the leucomonzogranite emplacement (291–287 Ma) that ended the batholith formation. In addition, there is no significant gap with the age of emplacement of the mafic tholeiitic magmatism (around 286 Ma) crosscutting the “A-type” granites. The late alkaline granites definitely do not show up here as precursors of the Tethyan rifting that began at about 170 Ma, i.e. some 100 Ma after their emplacement. It is thus necessary to examine if alternative hypotheses to the anorogenic model of the A-type Younger Granite province better fit the new geochronological data. A model involving depleted continental-crust derived magma should be compatible with the timing and geodynamical constraints as far as isotopic data are concerned.
OBJECTIVE:Cardiovascular (CV) events are more prevalent in chronic kidney disease (CKD) than in the general population, being the main cause of morbi-mortality. The aim of this project is to study ...the CV impact of the increase in the bioavailability of epoxyeicosatrienoic acids (EETs) by inhibiting soluble epoxide hydrolase (sEH) in the 5/6 nephrectomy model in 129/Sv mice.
DESIGN AND METHOD:5/6 Nx mice were treated with the sEH inhibitor t-AUCB, a diuretic (amiloride), or placebo for 10 weeks. Echocardiography was performed one week before the sacrifice to assess heart function. Hearts were weighed and histologic analyses were performed to evaluate fibrosis. Vascular function was studied ex vivo on the mesenteric artery. Sequential blood and urine tests were performed to assess kidney function.
RESULTS:Following 5/6 Nx, mice developed CKD. The CV consequences were heart hypertrophy (heart weight/tibia length Nx 5/6 vs. Sham7,9 ± 0,3 vs. 6,5 ± 0,5 mg/mm, p < 0,05), diastolic dysfunction (E/A ratioNx 5/6 vs. Sham1,29 ± 0,05 vs. 0,97 ± 0,04, p < 0,001) and diffuse fibrosis (Nx 5/6 vs. Sham57,8 ± 8,8 vs. 1,8 ± 1,8 %, p < 0,001). t-AUCB significantly prevented left ventricular hypertrophy (Nx 5/6 t-AUCB6,7 ± 0,2 mg/mm, p < 0,05 vs. Nx 5/6), diastolic dysfunction (Nx 5/6 t-AUCB1,12 ± 0,04 p < 0,05 vs. Nx 5/6), while beneficial effects on fibrosis were not significant (Nx 5/6 t-AUCB25,0 ± 12,7 % pns vs. Nx 5/6). No beneficial effects were present with amiloride. No vascular dysfunction was observed in our study.
CONCLUSIONS:Inhibition of sEH reduces the CV consequences associated with CKD. The beneficial effects related to the increase in the bioavailability of EETs hold a therapeutic potential in CKD patients.
Background: Autosomal dominant polycystic kidney disease is the most frequent hereditary kidney disease and is generally due to mutations in PKD1 and PKD2 , encoding polycystins 1 and 2. In autosomal ...dominant polycystic kidney disease, hypertension and cardiovascular disorders are highly prevalent, but their mechanisms are partially understood. Methods: Since endothelial cells express the polycystin complex, where it plays a central role in the mechanotransduction of blood flow, we generated a murine model with inducible deletion of Pkd1 in endothelial cells ( Cdh5-Cre ERT2 ; Pkd1 fl/fl ) to specifically determine the role of endothelial polycystin-1 in autosomal dominant polycystic kidney disease. Results: Endothelial deletion of Pkd1 induced endothelial dysfunction, as demonstrated by impaired flow-mediated dilatation of resistance arteries and impaired relaxation to acetylcholine, increased blood pressure and prevented the normal development of arteriovenous fistula. In experimental chronic kidney disease induced by subtotal nephrectomy, endothelial deletion of Pkd1 further aggravated endothelial dysfunction, vascular remodeling, and heart hypertrophy. Conclusions: Altogether, this study provides the first in vivo demonstration that specific deletion of Pkd1 in endothelial cells promotes endothelial dysfunction and hypertension, impairs arteriovenous fistula development, and potentiates the cardiovascular alterations associated with chronic kidney disease.
Heterozygous variants in RAR-related orphan receptor B (
) have recently been associated with susceptibility to idiopathic generalized epilepsy. However, few reports have been published so far ...describing pathogenic variants of this gene in patients with epilepsy and intellectual disability (ID). In this study, we aimed to delineate the epilepsy phenotype associated with
pathogenic variants and to provide arguments in favor of the pathogenicity of variants.
Through an international collaboration, we analyzed seizure characteristics, EEG data, and genotypes of a cohort of patients with heterozygous variants in
. To gain insight into disease mechanisms, we performed ex vivo cortical electroporation in mouse embryos of 5 selected variants, 2 truncating and 3 missense, and evaluated on expression and quantified changes in axonal morphology.
We identified 35 patients (17 male, median age 10 years, range 2.5-23 years) carrying 32 different heterozygous variants in
, including 28 single-nucleotide variants or small insertions/deletions (12 missense, 12 frameshift or nonsense, 2 splice-site variants, and 2 in-frame deletions), and 4 microdeletions; de novo in 18 patients and inherited in 10. Seizures were reported in 31/35 (89%) patients, with a median age at onset of 3 years (range 4 months-12 years). Absence seizures occurred in 25 patients with epilepsy (81%). Nineteen patients experienced a single seizure type: absences, myoclonic absences, or absences with eyelid myoclonia and focal seizures. Nine patients had absence seizures combined with other generalized seizure types. One patient had presented with absences associated with photosensitive occipital seizures. Three other patients had generalized tonic-clonic seizures without absences. ID of variable degree was observed in 85% of the patients. Expression studies in cultured neurons showed shorter axons for the 5 tested variants, both truncating and missense variants, supporting an impaired protein function.
In most patients, the phenotype of the
-related disorder associates absence seizures with mild-to-moderate ID. In silico and in vitro evaluation of the variants in our cohort, including axonal morphogenetic experiments in cultured neurons, supports their pathogenicity, showing a hypomorphic effect.
PDF
![[OP.4D.03] IMPACT OF THE IN...](http://api.summon.serialssolutions.com/2.0.0/image/issn/XR4PS5YY4K/0263-6352/small "[OP.4D.03] IMPACT OF THE INHIBITION OF SOLUBLE EPOXIDE HYDROLASE ON CARDIOVASCULAR CONSEQUENCES OF CHRONIC KIDNEY DISEASE")
