AMFR dysfunction causes autosomal recessive spastic paraplegia in human that is amenable to statin treatment in a preclinical model

Hereditary spastic paraplegias (HSP) are rare, inherited neurodegenerative or neurodevelopmental disorders that mainly present with lower limb spasticity and muscle weakness due to motor neuron dysfunction. Whole genome sequencing identified bi-allelic truncating variants in AMFR, encoding a RING-H2 finger E3 ubiquitin ligase anchored at the membrane of the endoplasmic reticulum (ER), in two previously genetically unexplained HSP-affected siblings. Subsequently, international collaboration recognized additional HSP-affected individuals with similar bi-allelic truncating AMFR variants, resulting in a cohort of 20 individuals from 8 unrelated, consanguineous families. Variants segregated with a phenotype of mainly pure but also complex HSP consisting of global developmental delay, mild intellectual disability, motor dysfunction, and progressive spasticity. Patient-derived fibroblasts, neural stem cells (NSCs), and in vivo zebrafish modeling were used to investigate pathomechanisms, including initial preclinical therapy assessment. The absence of AMFR disturbs lipid homeostasis, causing lipid droplet accumulation in NSCs and patient-derived fibroblasts which is rescued upon AMFR re-expression. Electron microscopy indicates ER morphology alterations in the absence of AMFR. Similar findings are seen in amfra-/- zebrafish larvae, in addition to altered touch-evoked escape response and defects in motor neuron branching, phenocopying the HSP observed in patients. Interestingly, administration of FDA-approved statins improves touch-evoked escape response and motor neuron branching defects in amfra-/- zebrafish larvae, suggesting potential therapeutic implications. Our genetic and functional studies identify bi-allelic truncating variants in AMFR as a cause of a novel autosomal recessive HSP by altering lipid metabolism, which may potentially be therapeutically modulated using precision medicine with statins

Investigation of pharmaceuticals in a conventional wastewater treatment plant: Removal efficiency, seasonal variation and impact of a nearby hospital

Discharges from the wastewater treatment plants (WWTPs) are among the main sources of contamination to receiving surface water, therefore the quality of treated wastewater needs to be properly monitored. However, not only the effluents of larger WWTPs employing advanced treatment processes have been considered, but also those from more conventional WWTPs. In this study, the occurrence and behavior of pharmaceuticals have been investigated in a conventional WWTP which receives wastewater from an urban area and a near-by hospital. 24-h composite samples were collected during one week before (influent wastewater, IWW) and after (effluent wastewater, EWW) treatment along three monitoring campaigns distributed over one year. Moreover, seven daily IWW samples discharged from a hospital were also collected. A preliminary wide-scope screening using liquid chromatography (LC) coupled to high resolution mass spectrometry allowed to identify a wide number of pharmaceuticals in the samples. Based on the screening findings, a list of 40 compounds was established for subsequent target quantitative analyses by LC-tandem mass spectrometry. Up to 75% of the compounds investigated were present in all wastewater samples. Analyte concentrations in hospital discharge samples were significantly higher, evidencing an important contribution in terms of pharmaceuticals content. Antibiotics showed the highest concentrations during the winter season, which could be related to the increase in the prescription of these compounds to treat respiratory infections. Data from this work show that the biological treatment applied was able to eliminate nearly half of the compounds under study, although still 12 pharmaceuticals were not or poorly removed.University Jaume I/[UJI-B2018-55]/UJI/EspañaMinistry of Science Innovation and University/[Ref RTI2018-097417-B-I00]//EspañaGeneralitat Valenciana/[Prometeo 2019/040]//EspañaUniversidad de Costa Rica/[OAICECAB-12-235-2016]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro en Investigación en Contaminación Ambiental (CICA