Evaluating LINE-1 methylation in cleft lip tissues and its association with early pregnancy exposures

Aim: To pilot investigation of methylation of long interspersed nucleotide element-1 in lip tissues from infants with nonsyndromic cleft lip, and its association with maternal periconceptional exposures. Methods: The lateral and medial sides of the cleft lips of 23 affected infants were analyzed for long interspersed nucleotide element-1 methylation by bisulfite conversion and pyrosequencing. Results: The medial side showed 1.8% higher methylation compared with the lateral side; p = 0.031, particularly in male infants (2.7% difference; p = 0.011) or when the mothers did not take folic acid during periconceptional period (2.4% difference; p = 0.011). These results were not statistically significant when Bonferroni adjustment was used. Conclusion: The observed differences in DNA methylation, although nonsignificant after correction for multiple comparisons, suggest that differential regulation of the two sides may impact lip fusion and warrant larger-scale replication

SARS-CoV-2 Breakthrough Infections: Incidence and Risk Factors in a Large European Multicentric Cohort of Health Workers

The research aimed to investigate the incidence of SARS-CoV-2 breakthrough infections and their determinants in a large European cohort of more than 60,000 health workers

Dolore neuropatico nelle neuropatie periferiche geneticamente determinate: qualità della vita, esercizio fisico e marcatori di neuroinfiammazione

Le neuropatie periferiche ereditarie demielinizzanti, assonali o miste, sono un gruppo eterogeneo di patologie a trasmissione genetica che coinvolgono i nervi periferici sia di tipo motorio che sensitivo interessando meno frequentemente il sistema nervoso autonomo e rappresentano circa il 30% delle neuropatie croniche. Tra queste, l’HMSN, anche nota con l’eponimo di malattia di Charcot-Marie-Tooth (CMT), è la più frequente con una prevalenza che risulta essere tra i 9-20 casi su 100.000 abitanti. La debolezza e l’amiotrofia distale, la deambulazione steppante, le deformità tipiche (piede cavo e dita ad artiglio) e i disturbi sensitivi sono le caratteristiche cliniche principali di questa neuropatia ereditaria. La disabilità, legata alla compromissione sensitivo-motoria, è variabile ed un’espressione importante del coinvolgimento sensitivo è il dolore neuropatico, presente frequentemente ma, spesso, trascurato. L’obiettivo primario della presente tesi è stato valutare l’incidenza del dolore cronico neuropatico in un gruppo di pazienti con diagnosi di malattia sensitivo-motoria ereditaria (HMSN), descriverne qualitativamente le caratteristiche ed approfondirne le correlazioni con vari aspetti quali la qualità della vita, la severità di malattia e l’attività fisica. Obiettivo secondario è stato anche quello di valutare il possibile ruolo che la neuroinfiammazione ha nella genesi e nel mantenimento del dolore, anche in relazione all’esercizio fisico. Sono stati inclusi 26 pazienti di età superiore ai 18 anni con diagnosi genetica confermata di malattia di Charcot-Marie-Tooth e seguiti presso il Centro per le Malattie Neuromuscolari del Dipartimento di Medicina Clinica e Sperimentale dell’Università di Pisa. I soggetti di entrambi i sessi, 13 maschi e 13 femmine, hanno un’età media di 48,5 anni (DS=15,4), un’età media alla diagnosi di 42,1 anni (DS=15,2) ed una durata media di sintomi di 11,7 anni (DS=6,6). A tutti i pazienti, dopo la valutazione clinico-elettrofisiologica, sono state somministrate scale per la valutazione del dolore, sia di tipo unidimensionale (Numerical Rating Scale-NRS) che di tipo multidimensionale (Douleur Neuropathique en 4 questions (DN4), Neuropathic Pain Symptom Inventory (NPSI) e McGill Pain Questionnaire (MPQ)) e scale che esplorano altri aspetti, quali la qualità di vita (SF-36 Health Survey), la fatica (Fatigue Severity Scale-FSS), la severità di malattia (Charcot-Marie Tooth Neuropathy Score-CMTNS) e l’attività fisica, valutata tramite un questionario sullo stile di vita e suddivisa, adattando una scala norvegese, in sedentaria(0h/settimana), bassa (<3h/settimana), moderata (3-5h/settimana) ed alta (≥6 h/settimana). In 15 pazienti è stato effettuato prelievo ematico da vena antecubitale del braccio per il dosaggio dei livelli ematici basali di citochine pro-infiammatorie, quali le interleuchine 1β e 6 (IL-β e IL-6) ed il fattore di necrosi tumorale (TNF-α), ed anti-infiammatorie, come l’interleuchina 4 (IL-4). L’utilizzo di questionari validati per il dolore neuropatico, in particolare il DN4 e l’NPSI, ha permesso di individuare i pazienti che riferiscono sintomi e segni sensitivi che, sulla base della letteratura, sono associati a tale condizione. Si è rilevato che il dolore, nel 73% dei casi, sia di tipo neuropatico senza alcuna differenza significativa di genere o tra i diversi sottotipi di CMT. Nel gruppo di pazienti con dolore neuropatico, l’età media, la durata di malattia e l’intensità del dolore sono risultati statisticamente maggiori. La sensazione dolorosa prevalentemente riportata dai pazienti è di tipo “scossa elettrica” in associazione a sintomi quali il formicolio e l’intorpidimento. Il dolore, inoltre, risulta essere profondo e a distribuzione simmetrica distale. Dall’analisi delle scale e dei questionari è emerso come il dolore si correli in maniera statisticamente significativa con il sintomo fatica, con la severità di malattia e con alcuni domini in cui è valutata la qualità di vita, soprattutto la limitazione delle attività specifiche svolte dal paziente per problemi fisici. Dal questionario sullo stile di vita, invece, non si sono evidenziate differenze significative nel livello di attività fisica svolta dai pazienti con e senza dolore neuropatico, facendo quindi ipotizzare che alla base di tale compromissione vi sia la patologia neuropatica nel suo complesso oltre che il dolore. Non si sono rilevate correlazioni significative fra il dolore, l’esercizio e l’infiammazione, valutati attraverso il dosaggio, dopo prelievo ematico, delle citochine IL-1β, TNF-α, IL-6 ed IL-4, risultato probabilmente dovuto alla bassa numerosità del campione (N=15). Il dolore neuropatico, quindi, è un sintomo frequente nei pazienti affetti da questa neuropatia ereditaria ed è associato ad una maggiore severità di malattia e ad una compromissione dei diversi domini in cui la qualità di vita viene valutata, soprattutto quelli che ne esplorano la dimensione “fisica”

Evaluation of hydroxyapatite distribution in a Poly-L-Lactic Acid (PLLA) scaffolds via Micro Computed Tomography (µCT)

Bone Tissue Engineering offers promising clinical alternative substitutes for bone defects, focusing on the use of polymer/ceramic composites. Hydroxyapatite (HA), a bioactive ceramic, has been implemented in bone substitution and regeneration due to its biocompatibility, osteoconductivity and close resemblance to the mineralized phase of human bone. Several techniques have been adopted to characterize composite scaffolds in terms of morphology, pore size and interconnection, filler content and distribution, but most of them are destructives. In this work, composite Poly-L-Lactic Acid (PLLA)-HA scaffolds (17.6 mm diameter and 35.7 mm height) were prepared via Thermally Induced Phase Separation (TIPS) by using a ternary PLLA-dioxane-water solution (polymer-solvent-nonsolvent). Two different concentrations (10% and 20% wt/wt filler/polymer ratio) of HA particles were chosen. The samples were characterized via Scanning Electron Microscopy (SEM) and Microcomputed Tomography (µCT), a non-invasive technique capable to analyze the internal structure of the sample. After the µCT reconstruction, an investigation of the distribution and average sizes of the filler was carried out through a 3D analysis software (CTAn). The obtained results demonstrated the possibility to analyze the internal distribution of the particles via a non-invasive technique; furthermore a homogeneous HA content and a uniform size distribution in the whole sample was observed, which allows one to assess that the scaffold production technique does not create neither a filler gradient nor a particle sedimentation

Influence of Nutritional Status and Physical Exercise on Immune Response in Metabolic Syndrome

Metabolic Syndrome (MetS) is a cluster of metabolic alterations mostly related to visceral adiposity, which in turn promotes glucose intolerance and a chronic systemic inflammatory state, characterized by immune cell infiltration. Such immune system activation increases the risk of severe disease subsequent to viral infections. Strong correlations between elevated body mass index (BMI), type-2-diabetes and increased risk of hospitalization after pandemic influenza H1N1 infection have been described. Similarly, a correlation between elevated blood glucose level and SARS-CoV-2 infection severity and mortality has been described, indicating MetS as an important predictor of clinical outcomes in patients with COVID-19. Adipose secretome, including two of the most abundant and well-studied adipokines, leptin and interleukin-6, is involved in the regulation of energy metabolism and obesity-related low-grade inflammation. Similarly, skeletal muscle hormones-called myokines-released in response to physical exercise affect both metabolic homeostasis and immune system function. Of note, several circulating hormones originate from both adipose tissue and skeletal muscle and display different functions, depending on the metabolic context. This review aims to summarize recent data in the field of exercise immunology, investigating the acute and chronic effects of exercise on myokines release and immune system function

Sexual dimorphism in the cerebrospinal fluid total protein content

Objectives Cerebrospinal fluid (CSF) is a clear, colorless body fluid filling the central nervous system. The determination of the CSF total protein (TP) content represents an important screening test of various pathologies. We aimed to address the effect of sex and age on CSF TP content and the use of the current upper reference limits (URLs). Methods CSF TP content was analysed in a selected population of 1,252 patients (648 women and 604 men; age 18-89 years) who underwent lumbar puncture as a part of the diagnostic work-up. Samples presenting (i) more than 5 white blood cells (WBC)/µL, (ii) discolorations and (iii) reduced glucose were not included. Results The CSF TP content​​ median values were significantly higher in men than in women (46 vs. 37&nbsp;mg/dL) even after adjusting for age and different hospital inpatients. CSF TP content positively correlated with age both in men and in women with a constant difference between sexes of 8.5&nbsp;mg/dL. Applying the most used URLs (mainly 45 and 50&nbsp;mg/dL, but also 60&nbsp;mg/dL), men received a laboratory report suggestive of altered CSF TP content more frequently than women. The use of age- and sex-calibrated CSF TP URLs reduced, but not eliminated, this sex-gap. Conclusions Using the current URLs, a condition of "elevated CSF TP content" may be overestimated in men or, conversely, underestimated in women, regardless of the age and of the diagnosis. These results highlighted the need to apply CSF TP URLs values ​​normalized for both sex and age

A Comprehensive Review of COVID-19-Related Olfactory Deficiency: Unraveling Associations with Neurocognitive Disorders and Magnetic Resonance Imaging Findings

Olfactory dysfunction (OD) is one of the most common symptoms in COVID-19 patients and can impact patients’ lives significantly. The aim of this review was to investigate the multifaceted impact of COVID-19 on the olfactory system and to provide an overview of magnetic resonance (MRI) findings and neurocognitive disorders in patients with COVID-19-related OD. Extensive searches were conducted across PubMed, Scopus, and Google Scholar until 5 December 2023. The included articles were 12 observational studies and 1 case report that assess structural changes in olfactory structures, highlighted through MRI, and 10 studies correlating the loss of smell with neurocognitive disorders or mood disorders in COVID-19 patients. MRI findings consistently indicate volumetric abnormalities, altered signal intensity of olfactory bulbs (OBs), and anomalies in the olfactory cortex among COVID-19 patients with persistent OD. The correlation between OD and neurocognitive deficits reveals associations with cognitive impairment, memory deficits, and persistent depressive symptoms. Treatment approaches, including olfactory training and pharmacological interventions, are discussed, emphasizing the need for sustained therapeutic interventions. This review points out several limitations in the current literature while exploring the intricate effects of COVID-19 on OD and its connection to cognitive deficits and mood disorders. The lack of objective olfactory measurements in some studies and potential validity issues in self-reports emphasize the need for cautious interpretation. Our research highlights the critical need for extensive studies with larger samples, proper controls, and objective measurements to deepen our understanding of COVID-19’s long-term effects on neurological and olfactory dysfunctions