Dipeptide repeat proteins inhibit homology-directed DNA double strand break repair in C9ORF72 ALS/FTD

BACKGROUND: The C9ORF72 hexanucleotide repeat expansion is the most common known genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two fatal age-related neurodegenerative diseases. The C9ORF72 expansion encodes five dipeptide repeat proteins (DPRs) that are produced through a non-canonical translation mechanism. Among the DPRs, proline-arginine (PR), glycine-arginine (GR), and glycine-alanine (GA) are the most neurotoxic and increase the frequency of DNA double strand breaks (DSBs). While the accumulation of these genotoxic lesions is increasingly recognized as a feature of disease, the mechanism(s) of DPR-mediated DNA damage are ill-defined and the effect of DPRs on the efficiency of each DNA DSB repair pathways has not been previously evaluated. METHODS AND RESULTS: Using DNA DSB repair assays, we evaluated the efficiency of specific repair pathways, and found that PR, GR and GA decrease the efficiency of non-homologous end joining (NHEJ), single strand annealing (SSA), and microhomology-mediated end joining (MMEJ), but not homologous recombination (HR). We found that PR inhibits DNA DSB repair, in part, by binding to the nucleolar protein nucleophosmin (NPM1). Depletion of NPM1 inhibited NHEJ and SSA, suggesting that NPM1 loss-of-function in PR expressing cells leads to impediments of both non-homologous and homology-directed DNA DSB repair pathways. By deleting NPM1 sub-cellular localization signals, we found that PR binds NPM1 regardless of the cellular compartment to which NPM1 was directed. Deletion of the NPM1 acidic loop motif, known to engage other arginine-rich proteins, abrogated PR and NPM1 binding. Using confocal and super-resolution immunofluorescence microscopy, we found that levels of RAD52, a component of the SSA repair machinery, were significantly increased iPSC neurons relative to isogenic controls in which the C9ORF72 expansion had been deleted using CRISPR/Cas9 genome editing. Western analysis of post-mortem brain tissues confirmed that RAD52 immunoreactivity is significantly increased in C9ALS/FTD samples as compared to controls. CONCLUSIONS: Collectively, we characterized the inhibitory effects of DPRs on key DNA DSB repair pathways, identified NPM1 as a facilitator of DNA repair that is inhibited by PR, and revealed deficits in homology-directed DNA DSB repair pathways as a novel feature of C9ORF72-related disease

A study on the characteristics of Algerian Hassi-Messaoud asphaltenes:Algerian Hassi-Messaoud asphaltenes: solubility and precipitation

This study focuses on detailed characterizations of asphaltene fractions extracted from the Algerian Hassi-Messaoud oil field. It was found that the extracted asphaltenes are not completely soluble in toluene, instead two fractions of asphaltenes were obtained upon solubilizing the heptane-precipitated neat asphaltenes in toluene. Extensive characterizations of the toluene-soluble and insoluble fractions were carried out using elemental analysis, Fourier transform infrared (FTIR), thermogravimetric analysis (TGA), X-ray diffraction (XRD) and solid-state nuclear magnetic resonance (ssNMR). It was suggested that the high oxygen content and uneven compositional structures are the main contributors to asphaltene instability. The toluene-insoluble fractions were found to have higher polarity and aromaticity as well as more oxygen content than the neat asphaltenes and toluene-soluble fractions

The spectrum of MEFV gene mutations and genotype-phenotype correlation in Egyptian patients with familial Mediterranean fever

Background: Familial Mediterranean fever (FMF) is an autosomal recessive disease mainly affecting subjects of the Mediterranean origin. It is an auto-inflammatory periodic disorder that is caused by mutations in the Mediterranean fever gene (MEFV) located on chromosome 16.Methods: The current study was designed to assess the prevalence and frequency of different MEFV gene mutations among 104 FMF clinically diagnosed Egyptian patients and to evaluate the change extent in the values of some biochemical markers (ESR, CRP, Fibrinogen-C, SAA and IL1) in different participants with different FMF severity scores.Results: According to allele status 28 patients (27%) were homozygous mutation carriers, 38 (36.5%) were with compound heterozygous mutations and 38 (36.5%) were identified as heterozygous for one of the studied mutations. Of the studied mutations, M694I, E148Q, V726A, M680I, and M694V accounted for 28.1%, 26.8%, 16.9%, and 11.3% of mutations respectively. The R761H and P369S mutations were rarely encountered mutations (1.4%). The clinical features with M694I were associated with more severe clinical course. There is a drastic elevation in the levels of estimated parameters as their levels were increased as long as the severity of the disease increased.Conclusions: The diagnosis of FMF cannot be performed on the basis of genetic testing or clinical criteria alone. So, we recommended the combination between clinical and molecular profiling for FMF diagnosis and scoring

Comparative evaluation of the material of the artificial levees : A case study along the Tisza and Maros Rivers, Hungary

Artificial levees have major importance in protecting human livesand infrastructure as they are essential elements of the flood protectionmeasures. Nevertheless, the lack of the necessary information about theirstructure and internal composition might cause high risks. To monitor theirstability, integrated surveys are needed, including geophysical andgeotechnical methods. Levees along the rivers in Hungary were constructedmore than 150 years ago, and they were heightened several times; therefore,investigations are required to assure their performance in flood riskmitigation. Our investigation aimed to utilise non-invasive geophysicaltechniques, primarily electrical resistivity imaging, with the validationof geotechnical investigations to map and compare the compositional andstructural variations of two very different levee sections along RiverTisza and River Maros. Integrating the analysed drilling data with ERTprofiles showed that the main composition of the investigated Tisza leveesection is fine and medium silt with an average resistivity 30 Ωm, however,the investigated section of Maros levee was built of not only of fine andmedium silt but also of medium and coarse sand exhibiting higherresistivity values reaching up to 2200 Ωm. Several physical parameters weremeasured to study the nature of constituting levee materials like moisturecontent, grain-size, porosity, bulk-density, saturated hydraulicconductivity, and resistivity. It was found that most of them show aconnection with resistivity, but the hydraulic conductivity did not show adirect connection, however the latter could exhibit the aquitard nature ofTisza levee materials and the non-aquitard nature of Maros levee materials

FTD/ALS-associated poly(GR) protein impairs the Notch pathway and is recruited by poly(GA) into cytoplasmic inclusions

C9ORF72 repeat expansion is the most common genetic mutation in frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). Abnormal dipeptide repeat proteins (DPRs) generated from repeat-associated non-AUG (RAN) translation of repeat-containing RNAs are thought to be pathogenic; however, the mechanisms are unknown. Here we report that (GR)80 and (PR)80 are toxic in neuronal and non-neuronal cells in Drosophila. In contrast to reported shorter poly(GR) forms, (GR)80 is mostly localized throughout the cytosol without detectable accumulation in the nucleolus, accompanied by suppression of Notch signaling and cell loss in the wing. Some Notch target genes are also downregulated in brains and iPSC-derived cortical neurons of C9ORF72 patients. Increased Notch expression largely suppressed (GR)80-induced cell loss in the wing. When co-expressed in Drosophila, HeLa cells, or human neurons, (GA)80 recruited (GR)80 into cytoplasmic inclusions, partially decreasing the toxicity of (GR)80 and restoring Notch signaling in Drosophila. Thus, different DPRs have opposing roles in cell loss and we identify the Notch pathway as one of the receptor signaling pathways that might be compromised in C9ORF72 FTD/ALS

Prevalence of Body Dysmorphic Disorder (BDD) among the Lebanese University students: Associated risk factors and repercussion on mental health

Background: Body Dysmorphic Disorder (BDD) is a psychological illness characterized by persistent and intrusive preoccupation with an imagined or slight defect in appearance. For individuals with BDD, these perceived defects are significant and prominent, leading to emotional distress, depression, anxiety, and impairment in daily functioning. Despite its impact, no prior studies have explored the prevalence of BDD among Lebanese university students. Objectives: This study aimed to determine the prevalence of BDD and investigate its association with mental health status (depression and anxiety), religiosity, eating disorder risk, and self-esteem among Lebanese University students.  Methodology: A cross-sectional study was conducted in March 2020, involving students from the Lebanese University. Data were collected using the dysmorphic concern questionnaire (DCQ), Rosenberg self-esteem scale (RSE), religiosity scale, patient health questionnaire (PHQ-9), generalized anxiety disorder assessment (GAD-7), Eating Attitude test-26 scale (EAT-26). Results: A total of 6448 participants were enrolled in our study. The overall prevalence of BDD among university students was (6.4%).Our results showed that Lebanese students with BDD were more likely to have anxiety ORadj 1.3 (95% CI:1.2-1.7) p-value 0.001, depression ORadj 1.2 (95% C:1.15-1.5) p-value 0.007, and eating disorder (Bulimia & Food preoccupation ORadj 1.06 (95% CI:1.03-1.2) P-value 0.0, and oral control ORadj 1.09 (95% CI:1.05-1.1)  P-value < 0.001) compared to those with no BDD. We also found that students with BDD had Lower Self-esteem ORadj 0.88 (95%CI: 0.78-0.9) P-value <0.001), and less likely to be religious ORadj 0.88 (95%CI: 0.82-0.9) P-value 0.02) compared to those with no BDD. Conclusion: This pioneering study in Lebanon sheds light on the prevalence of BDD among Lebanese university students and its associated factors. BDD was found to be significantly linked to elevated levels of depression, anxiety, and eating disorders, along with lower levels of self-esteem and religiosity. These findings underscore the importance of early detection of BDD during adolescence and young adulthood, necessitating timely psychological intervention to prevent chronicity and complications. Moreover, promoting widespread awareness through various social media and public platforms is essential to address this psychological issue effectively