The Ribosomal Protein RpL22 Interacts In Vitro with 5′-UTR Sequences Found in Some Drosophila melanogaster Transposons

Mobility of eukaryotic transposable elements (TEs) are finely regulated to avoid an excessive mutational load caused by their movement. The transposition of retrotransposons is usually regulated through the interaction of host- and TE-encoded proteins, with non-coding regions (LTR and 5′-UTR) of the transposon. Examples of new potent cis-acting sequences, identified and characterized in the non-coding regions of retrotransposons, include the insulator of gypsy and Idefix, and the enhancer of ZAM of Drosophila melanogaster. Recently we have shown that in the 5′-UTR of the LTR-retrotransposon ZAM there is a sequence structured in tandem-repeat capable of operating as an insulator both in Drosophila (S2R+) and human cells (HEK293). Here, we test the hypothesis that tandem repeated 5′-UTR of a different LTR-retrotransposon could accommodate similar regulatory elements. The comparison of the 5′-UTR of some LTR-transposons allowed us to identify a shared motif of 13bp, called Transposable Element Redundant Motif (TERM). Surprisingly, we demonstrated, by Yeast One-Hybrid assay, that TERM interacts with the D. melanogaster ribosomal protein RpL22. The Drosophila RpL22 has additional Ala-, Lys- and Prorich sequences at the amino terminus, which resembles the carboxy-terminal portion of histone H1 and histone H5. For this reason, it has been hypothesized that RpL22 might have two functions, namely the role in organizing the ribosome, and a potential regulatory role involving DNA-binding similar to histone H1, which represses transcription in Drosophila. In this paper, we show, by two independent sets of experiments, that DmRpL22 is able to directly and specifically bind DNA of Drosophila melanogaster

Metabolomic fingerprinting of renal disease progression in Bardet-Biedl syndrome reveals mitochondrial dysfunction in kidney tubular cells.

Chronic kidney disease (CKD) is a major clinical sign of patients with Bardet-Biedl syndrome (BBS), especially in those carrying BBS10 mutations. Twenty-nine patients with BBS and 30 controls underwent a serum-targeted metabolomic analysis. In vitro studies were conducted in two kidney-derived epithelial cell lines, where Bbs10 was stably deleted (IMCD3-Bbs10-/-cells) and over-expressed. The CKD status affected plasmatic metabolite fingerprinting in both patients with BBS and controls. Specific phosphatidylcholine and acylcarnitines discriminated eGFR decline only in patients with BBS. IMCD3-Bbs10-/ cells displayed intracellular lipidaccumulation, reduced mitochondrial potential membrane and citrate synthase staining. Mass-Spectrometry-based analysis revealed that human BBS10 interacted with six mitochondrial proteins, in vitro. In conclusion, renal dysfunction correlated with abnormal phosphatidylcholine and acylcarnitines plasma levels in patients with BBS; in vitro, Bbs10 depletion caused mitochondrial defects while human BBS10 interacted with several mitochondria-related proteins, suggesting an unexplored role of this protein

Nephroplex: a kidney-focused NGS panel highlights the challenges of PKD1 sequencing and identifies a founder BBS4 mutation

Background: Genetic testing of patients with inherited kidney diseases has emerged as a tool of clinical utility by improving the patients' diagnosis, prognosis, surveillance and therapy. Methods: The present study applied a Next Generation Sequencing (NGS)-based panel, named NephroPlex, testing 115 genes causing renal diseases, to 119 individuals, including 107 probands and 12 relatives. Thirty-five (poly)cystic and 72 non (poly)cystic individuals were enrolled. The latter subgroup of patients included Bardet-Biedl syndrome (BBS) patients, as major components. Results: Disease-causing mutations were identified in 51.5 and 40% of polycystic and non-polycystic individuals, respectively. Autosomal dominant polycystic kidney disease (ADPKD) patients with truncating PKD1 variants showed a trend towards a greater slope of the age-estimated glomerular filtration rate (eGFR) regression line than patients with (i) missense variants, (ii) any PKD2 mutations and (iii) no detected mutations, according to previous findings. The analysis of BBS individuals showed a similar frequency of BBS4,9,10 and 12 mutations. Of note, all BBS4-mutated patients harbored the novel c.332+1G>GTT variant, which was absent in public databases, however, in our internal database, an additional heterozygote carrier was found. All BBS4-mutated individuals originated from the same geographical area encompassing the coastal provinces of Naples. Discussion: In conclusion, these findings indicate the potential for a genetic panel to provide useful information at both clinical and epidemiological levels

Impact of COVID-19 Pandemic on Children and Adolescents with Neuropsychiatric Disorders: Emotional/Behavioral Symptoms and Parental Stress

The objective of our study was to evaluate the impact of the COVID-19 pandemic on the emotional and behavioral symptoms in minors with neuropsychiatric disorders and on parental stress through a standardized neuropsychological assessment, comparing the data collected before the pandemic with those collected during the lock-down. Another goal of our study was to analyze the relationship between parental stress and behavioral/emotional symptoms in children. Our study was conducted on 383 families of patients who had already been referred at the Child Neuropsychiatry Unit of the University Hospital of Salerno for different neuropsychiatric conditions. All the parents completed two neuropsychological standardized questionnaires for the assessment of parental stress (PSI—Parenting Stress Index-Short Form) and the emotional/behavioral problems of their children (Child Behaviour CheckList). The data collected during the pandemic were compared with those collected from questionnaires administered during the six months preceding the pandemic, as is our usual clinical practice. The comparison between the mean scores of PSI and CBCL before and after the pandemic showed a statistically significant increase in all subscales analyzed in the total sample. The correlation analysis showed significant positive relationship between the subscale Total Stress of PSI and the subscales Total Problems and Internalizing Problems of CBCL. Our study suggested that the COVID-19 pandemic and the corresponding measures adopted led to an increase in internalizing and externalizing symptoms in children and adolescents with neuropsychiatric disorder. Similarly, parental stress increased during COVID-19 and ahigher level of stress in parents can be related to the internalizing symptoms of their children

Generation of Induced Pluripotent Stem Cells from Patients with Duchenne Muscular Dystrophy and their induction to Neurons

Duchenne muscular dystrophy (DMD) is an X-linked recessive disease characterized by deficient expression of the cytoskeletal protein dystrophin. DMD has been associated with intellectual disability and mental retardation (MR) and is present in about a third of all patients. Loss of Dp71, the major dystrophin-gene product in brain, and the dystrophin associated proteins (DAPs) are thought to contribute to severity of MR, but the specific function of the neural dystrophin proteins are poorly understood for a limited access to DMD patients brain tissue (1). Differentiation of induced Pluripotent Stem Cells (iPSCs) provides an opportunity to generate an unlimited supply of living neurons genetically identical to those present in patients. In this study we obtained DMD-iPSCs from peripheral blood mononuclear cells of DMD patients with cognitive impairment and we performed morphological (fluorescence and electron microscopy), molecular (Western Blot and Real Time PCR) and functional (electrophysiology) characterization both of iPSC-derived Neural Stem Cells (NSCs) and the differentiated neurons. Preliminary data showed a reduction of Dp71 and DAPs proteins, including the AQP4, potassium channel Kir4.1, α- and β-dystroglycan (α/βDG) and α-syntrophin (αSyn), both at transcriptional and traductional level, coupled with membrane dys-arrangment in DMD-iPSCs compared with healthy iPSCs. Moreover, we demonstrated that the neurons obtained from the differentiation of iPSCs derived from DMD patient showed after confocal analysis, altered cytoskeleton and reduction in Dp71expression, and by single-cell imaging experiments and electrophysiology, altered intracellular calcium homeostasis, in analogy with what shown in the dystrophic mdx mouse neurons (2). Overall these results showed that the Dp71 and DAPs alterations affect also the neural precursor as well as the differentiated neurons in DMD patients, so suggesting a key role in the pathogenesis of neurocognitive deficits in DMD disease

Reliability of the Performance of Upper Limb assessment in Duchenne muscular dystrophy

Abstract The Performance of Upper Limb was specifically designed to assess upper limb function in Duchenne muscular dystrophy. The aim of this study was to assess (1) a cohort of typically developing children from the age of 3 years onwards in order to identify the age when the activities assessed in the individual items are consistently achieved, and (2) a cohort of 322 Duchenne children and young adults to establish the range of findings at different ages. We collected normative data for the scale validation on 277 typically developing subjects from 3 to 25 years old. A full score was consistently achieved by the age of 5 years. In the Duchenne cohort there was early involvement of the proximal muscles and a proximal to distal progressive involvement. The scale was capable of measuring small distal movements, related to activities of daily living, even in the oldest and weakest patients. Our data suggest that the assessment can be reliably used in both ambulant and non ambulant Duchenne patients in a multicentric setting and could therefore be considered as an outcome measure for future trials

BIOMASS: ENVIRONMENTAL COMPATIBILITY AND STRATEGIES FOR ENERGY PLANNING

1 ABSTRACT Authorities and agencies in many nations are discussing ambitious targets in order to mitigate the most dangerous impact of human activity (climate change) on the environment; by considering health of human society, and also concerns about economics aspects, some regions have already set reduction targets. The European Union has set itself the ambitious target to increase the share of renewable sources in final energy consumption to 20 % by 2020, in order to reduce both GHGs emissions and reliance on fossil fuels from foreign markets. In the framework of this productive change, biomass (one of the major sources of renewable energy) plants are getting a lot of attention, but it is necessary to determine whether, in comparison with individuated climate and resource benefits, the use of this type of energy is also environmentally beneficial. A complete assessment of the impact of a biomass energy plant requires the evaluation of many different aspects at local and global scales. From one side there are important positive aspect such as limitation of greenhouse gas (GHG) emission, use of secondary materials, improvement of local activities, right destination of organic sludges, absence of toxicity, and limited cost; from the other side, the effects on land use, need of chemicals and imported resources, expansion and/or intensification of agriculture in other locations, and, chiefly, formation of secondary pollutants (dust, nitrogen oxides, and VOC) must be carefully considered. The present thesis has the aim to evaluate all these aspects, by providing a methodological approach and a scheme, to be applied for practical application in real situations and in order to perform a quantitative analysis of the environmental impacts. The proposed methodology permits to evaluate the environmental compatibility of a specific biomass plant and the potential energy production from biomass at regional level. Some specific numerical applications, referred to structure of plants and local impact assessment, and also to planning considerations utilized to define the bioenergy potentials and environmental consequences in Italian regions of different structure. The overall description of technological schemes and process and territorial acceptability of bioenergy plants has been completed with considerations about production of biogas and biomethane, and fermentation of low-cost natural materials to produce second generation biofuels. The obtained conclusions, that with their numerical definition can be considered very useful for the specific studied situations, can also be considered fairly representative as a methodological approach, with the general purpose to study with a global approach the effects of biomass energy plants

BIOMASS: ENVIRONMENTAL COMPATIBILITY AND STRATEGIES FOR ENERGY PLANNING

1 ABSTRACT Authorities and agencies in many nations are discussing ambitious targets in order to mitigate the most dangerous impact of human activity (climate change) on the environment; by considering health of human society, and also concerns about economics aspects, some regions have already set reduction targets. The European Union has set itself the ambitious target to increase the share of renewable sources in final energy consumption to 20 % by 2020, in order to reduce both GHGs emissions and reliance on fossil fuels from foreign markets. In the framework of this productive change, biomass (one of the major sources of renewable energy) plants are getting a lot of attention, but it is necessary to determine whether, in comparison with individuated climate and resource benefits, the use of this type of energy is also environmentally beneficial. A complete assessment of the impact of a biomass energy plant requires the evaluation of many different aspects at local and global scales. From one side there are important positive aspect such as limitation of greenhouse gas (GHG) emission, use of secondary materials, improvement of local activities, right destination of organic sludges, absence of toxicity, and limited cost; from the other side, the effects on land use, need of chemicals and imported resources, expansion and/or intensification of agriculture in other locations, and, chiefly, formation of secondary pollutants (dust, nitrogen oxides, and VOC) must be carefully considered. The present thesis has the aim to evaluate all these aspects, by providing a methodological approach and a scheme, to be applied for practical application in real situations and in order to perform a quantitative analysis of the environmental impacts. The proposed methodology permits to evaluate the environmental compatibility of a specific biomass plant and the potential energy production from biomass at regional level. Some specific numerical applications, referred to structure of plants and local impact assessment, and also to planning considerations utilized to define the bioenergy potentials and environmental consequences in Italian regions of different structure. The overall description of technological schemes and process and territorial acceptability of bioenergy plants has been completed with considerations about production of biogas and biomethane, and fermentation of low-cost natural materials to produce second generation biofuels. The obtained conclusions, that with their numerical definition can be considered very useful for the specific studied situations, can also be considered fairly representative as a methodological approach, with the general purpose to study with a global approach the effects of biomass energy plants