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Introduction: Seasonal influenza generally represents an underestimated public health problem with significant socioeconomic implications. Monitoring and detecting influenza epidemics are important tasks that require integrated strategies. Wastewater-based epidemiology (WBE) is an emerging field that uses wastewater data to monitor the spread of disease and assess the health of a community. It can represent an integrative surveillance tool for better understanding the epidemiology of influenza and prevention strategies in public health. Methods: We conducted a study that detected the presence of Influenza virus RNA using a wastewater-based approach. Samples were collected from five wastewater treatment plants in five different municipalities, serving a cumulative population of 555,673 Sicilian inhabitants in Italy. We used the RT-qPCR test to compare the combined weekly average of Influenza A and B viral RNA in wastewater samples with the average weekly incidence of Influenza-like illness (ILI) obtained from the Italian national Influenza surveillance system. We also compared the number of positive Influenza swabs with the viral RNA loads detected from wastewater. Our study investigated 189 wastewater samples. Results: Cumulative ILI cases substantially overlapped with the Influenza RNA load from wastewater samples. Influenza viral RNA trends in wastewater samples were similar to the rise of ILI cases in the population. Therefore, wastewater surveillance confirmed the co-circulation of Influenza A and B viruses during the season 2022/2023, with a similar trend to that reported for the weekly clinically confirmed cases. Conclusion: Wastewater-based epidemiology does not replace traditional epidemiological surveillance methods, such as laboratory testing of samples from infected individuals. However, it can be a valuable complement to obtaining additional information on the incidence of influenza in the population and preventing its spread.
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Vírus da Influenza A , Influenza Humana , Águas Residuárias , Sicília/epidemiologia , Humanos , Influenza Humana/epidemiologia , Influenza Humana/virologia , Águas Residuárias/virologia , Vírus da Influenza A/isolamento & purificação , Vírus da Influenza A/genética , Estações do Ano , Vírus da Influenza B/isolamento & purificação , Vírus da Influenza B/genética , RNA Viral/análise , Cidades/epidemiologiaRESUMO
BACKGROUND: Antimicrobial resistance (AMR) is a topic of concern, especially in high-level care departments like neonatal intensive care units (NICUs). The systematic use of an "active" epidemiological surveillance system allows us to observe and analyze any changes in microbial distribution, limiting the risk of healthcare-associated infection (HAI) development. METHODS: We have conducted a longitudinal observational study in the five NICUs of Palermo, comparing the "pre-pandemic period" (March 2014-February 2020) with the "pandemic" one (March 2020-February 2022). The primary aim of the study was to evaluate the cumulative prevalence of carriage from multi-drug resistant (MDR) bacteria in the cumulative NICUs (NICU C). RESULTS: During the "pre-pandemic period", 9407 swabs were collected (4707 rectal, 4700 nasal); on the contrary, during the "pandemic period", a total of 2687 swabs were collected (1345 rectal, 1342 nasal). A statistically significant decrease in MDR-Gram-negative bacteria (GNB) carriage prevalence was detected during the pandemic. At the same time, there was a general worsening of the carriage of carbapenemase-forming MDR-GNB (CARBA-R+) and methicillin-resistant Staphylococcus aureus (MRSA) during the pandemic period. A significant reduction in methicillin-susceptible Staphylococcus aureus (MSSA) carriage was detected too. CONCLUSIONS: The surveillance of MDRO carriage in NICUs is fundamental for limiting the social and economic burden of HAIs.
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Long COVID is a recognized post-viral syndrome characterized by neurological, somatic and neuropsychiatric symptoms that might last for long time after SARS-CoV-2 infection. An ever-growing number of patients come to the observation of General Practitioners complaining of mild or moderate symptoms after the resolution of the acute infection. Nine General Practitioners from the Rome area (Italy) performed a retrospective analysis in order to evaluate the role of the supplementation with Palmitoylethanolamide co-ultramicronized with Luteolin (PEALUT) on neurologic and clinical symptoms reported by their patients after COVID-19 resolution. Supplementation with PEALUT helped to improve all patient-reported symptoms, especially pain, anxiety and depression, fatigue, brain fog, anosmia and dysgeusia, leading to an overall improvement in patients' health status. To our knowledge these are the first data presented on Long COVID patients collected in a territorial setting. Despite their preliminary nature, these results highlight the pathogenetic role of "non-resolving" neuroinflammation in Long COVID development and consequently the importance of its control in the resolution of the pathology and put the focus on the General Practitioner as the primary figure for early detection and management of Long COVID syndrome in a real-life setting. Future randomized, controlled, perspective clinical trials are needed to confirm this preliminary observation.
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COVID-19 , Clínicos Gerais , Humanos , Síndrome de COVID-19 Pós-Aguda , Luteolina , Estudos Retrospectivos , SARS-CoV-2RESUMO
TG2576 mice show highest levels of the full length mutant Swedish Human Amyloid Precursor Protein (APPKM670/671LN) during prodromal and early sympotomatic stages. Interestingly, this occurs in association with the unbalanced expression of two of its RNA Binding proteins (RBPs) opposite regulators, the Fragile-X Mental Retardation Protein (FMRP) and the heteronuclear Ribonucleoprotein C (hnRNP C). Whether an augmentation in overall translational efficiency also contributes to the elevation of APP levels at those early developmental stages is currently unknown. We investigated this possibility by performing a longitudinal polyribosome profiling analysis of APP mRNA and protein in total hippocampal extracts from Tg2576 mice. Results showed that protein polysomal signals were exclusively detected in pre-symptomatic (1 months) and early symptomatic (3 months) mutant mice. Differently, hAPP mRNA polysomal signals were detected at any age, but a peak of expression was found when mice were 3-month old. Consistent with an early but transient rise of translational efficiency, the phosphorylated form of the initial translation factor eIF2α (p-eIF2α) was reduced at pre-symptomatic and early symptomatic stages, whereas it was increased at the fully symptomatic stage. Pharmacological downregulation of overall translation in early symptomatic mutants was then found to reduce hippocampal levels of full length APP, Aßspecies, BACE1 and Caspase-3, to rescue predominant LTD at hippocampal synapses, to revert dendritic spine loss and memory alterations, and to reinstate memory-induced c-fosactivation. Altogether, our findings demonstrate that overall translation is upregulated in prodromal and early symptomatic Tg2576 mice, and that restoring proper translational control at the onset of AD-like symptoms blocks the emergence of the AD-like phenotype.
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Secretases da Proteína Precursora do Amiloide/metabolismo , Precursor de Proteína beta-Amiloide/metabolismo , Sintomas Prodrômicos , Regulação para Cima , Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides , Animais , Modelos Animais de Doenças , Fator de Iniciação 2 em Eucariotos/metabolismo , Feminino , Proteína do X Frágil da Deficiência Intelectual , Hipocampo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurônios/metabolismo , Fosforilação , RNA Mensageiro/metabolismo , Sinapses/metabolismoRESUMO
During development, ribosome biogenesis and translation reach peak activities, due to impetuous cell proliferation. Current models predict that protein synthesis elevation is controlled by transcription factors and signalling pathways. Developmental models addressing translation factors overexpression effects are lacking. Eukaryotic Initiation Factor 6 (eIF6) is necessary for ribosome biogenesis and efficient translation. eIF6 is a single gene, conserved from yeasts to mammals, suggesting a tight regulation need. We generated a Drosophila melanogaster model of eIF6 upregulation, leading to a boost in general translation and the shut-down of the ecdysone biosynthetic pathway. Indeed, translation modulation in S2 cells showed that translational rate and ecdysone biosynthesis are inversely correlated. In vivo, eIF6-driven alterations delayed Programmed Cell Death (PCD), resulting in aberrant phenotypes, partially rescued by ecdysone administration. Our data show that eIF6 triggers a translation program with far-reaching effects on metabolism and development, stressing the driving and central role of translation.
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Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Ecdisona/biossíntese , Regulação da Expressão Gênica no Desenvolvimento , Fatores de Iniciação de Peptídeos/genética , Biossíntese de Proteínas/genética , Animais , Animais Geneticamente Modificados , Apoptose/genética , Linhagem Celular , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/metabolismo , Discos Imaginais/crescimento & desenvolvimento , Discos Imaginais/metabolismo , Fatores de Iniciação de Peptídeos/metabolismo , Ribossomos/genética , Ribossomos/metabolismo , Transdução de Sinais/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
TDP-43 is a well known RNA binding protein involved in the pathogenesis of Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Lobar Dementia (FTLD). In physiological conditions, TDP-43 mainly localizes in the nucleus and shuttles, at least in neurons, to the cytoplasm to form TDP-43 RNA granules. In the nucleus, TDP-43 participates to the expression and splicing of RNAs, while in the cytoplasm its functions range from transport to translation of specific mRNAs. However, if loss or gain of these TDP-43 functions are affected in ALS/FTLD pathogenesis is not clear. Here, we report that TDP-43 localizes on ribosomes not only in primary neurons but also in SH-SY5Y human neuroblastoma cells. We find that binding of TDP-43 to the translational machinery is mediated by an interaction with a specific ribosomal protein, RACK1, and that an increase in cytoplasmic TDP-43 represses global protein synthesis, an effect which is rescued by overexpression of RACK1. Ribosomal loss of RACK1, which excludes TDP-43 from the translational machinery, remarkably reduces formation of TDP-43 cytoplasmic inclusions in neuroblastoma cells. Finally, we corroborate the interaction between TDP-43 and RACK1 on polyribosomes of neuroblastoma cells with mis-localization of RACK1 on TDP-43 positive cytoplasmic inclusions in motor neurons of ALS patients. In conclusions, results from this study suggest that TDP-43 represents a translational repressor not only for specific mRNAs but for overall translation and that its binding to polyribosomes through RACK1 may promote, under conditions inducing ALS pathogenesis, the formation of cytoplasmic inclusions.
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Esclerose Lateral Amiotrófica/genética , Proteínas de Ligação a DNA/biossíntese , Demência Frontotemporal/genética , Proteínas de Ligação ao GTP/biossíntese , Proteínas de Neoplasias/biossíntese , Receptores de Superfície Celular/biossíntese , Esclerose Lateral Amiotrófica/patologia , Núcleo Celular/genética , Citoplasma/genética , Proteínas de Ligação a DNA/genética , Demência Frontotemporal/patologia , Proteínas de Ligação ao GTP/genética , Regulação da Expressão Gênica , Humanos , Neurônios Motores/metabolismo , Neurônios Motores/patologia , Mutação , Proteínas de Neoplasias/genética , Neuroblastoma , Polirribossomos/genética , Biossíntese de Proteínas/genética , Splicing de RNA/genética , RNA Mensageiro/biossíntese , Receptores de Quinase C Ativada , Receptores de Superfície Celular/genética , Ribossomos/genéticaRESUMO
Ribosomopathies are a family of inherited disorders caused by mutations in genes necessary for ribosomal function. Shwachman-Diamond Bodian Syndrome (SDS) is an autosomal recessive disease caused, in most patients, by mutations of the SBDS gene. SBDS is a protein required for the maturation of 60S ribosomes. SDS patients present exocrine pancreatic insufficiency, neutropenia, chronic infections, and skeletal abnormalities. Later in life, patients are prone to myelodisplastic syndrome and acute myeloid leukemia (AML). It is unknown why patients develop AML and which cellular alterations are directly due to the loss of the SBDS protein. Here we derived mouse embryonic fibroblast lines from an SbdsR126T/R126T mouse model. After their immortalization, we reconstituted them by adding wild type Sbds. We then performed a comprehensive analysis of cellular functions including colony formation, translational and transcriptional RNA-seq, stress and drug sensitivity. We show that: 1. Mutant Sbds causes a reduction in cellular clonogenic capability and oncogene-induced transformation. 2. Mutant Sbds causes a marked increase in immature 60S subunits, limited impact on mRNA specific initiation of translation, but reduced global protein synthesis capability. 3. Chronic loss of SBDS activity leads to a rewiring of gene expression with reduced ribosomal capability, but increased lysosomal and catabolic activity. 4. Consistently with the gene signature, we found that SBDS loss causes a reduction in ATP and lactate levels, and increased susceptibility to DNA damage. Combining our data, we conclude that a cell-specific fragile phenotype occurs when SBDS protein drops below a threshold level, and propose a new interpretation of the disease.
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Homeostase , Fenótipo , Proteínas/genética , Subunidades Ribossômicas Maiores de Eucariotos/genética , Trifosfato de Adenosina/metabolismo , Animais , Linhagem Celular , Transformação Celular Neoplásica , Dano ao DNA , Fibroblastos/metabolismo , Ácido Láctico/metabolismo , Camundongos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Subunidades Ribossômicas Maiores de Eucariotos/metabolismoRESUMO
To detect central neuron activation, expression of the transcription factor Fos and phosphorylation of the protein kinase ERK (pERK) can be visualized by immunocytochemistry. These approaches have been extensively used to quantify the activation of nociceptive neurons in the spinal dorsal horn (DH) following peripheral stimulation in vivo. Here we propose an alternative and simplified in vitro model to investigate Fos and pERK expression based on the stimulation of acutely dissected spinal cord slices to mimic acute inflammatory changes in DH. Transverse slices were obtained from postnatal (P8-P12) CD1 mice and were treated for 5 min with capsaicin (CAP, 2 µM). CAP induces a strong release of glutamate from primary afferent terminals which, in turn, excites spinal DH neurons. Since ERK phosphorylation and Fos expression occur following different time frames, two distinct protocols were used to detect their activation. Thus, for studying Fos immunoreactivity CAP-treated slices were left for 3h in Krebs solution after stimulation. Instead, for studying pERK immunoreactivity slices were maintained in Krebs solution for only 15 min after stimulation. Both Fos and pERK were significantly up-regulated following CAP challenge. To validate our model we tested the efficacy of octreotide (OCT, 1 µM) in preventing the CAP effect on Fos and pERK expression. OCT is a synthetic antinociceptive analog of somatostatin, one of the neuropeptides involved in the negative modulation of pain signals in DH. After CAP, OCT reduced the response to both Fos and pERK. Our data validate the use of Fos and pERK immunoreactivity in vitro to investigate the activation of spinal nociceptive pathways and testing potentially antinociceptive molecules.