Extracellular vesicles (EVs), comprising exosomes and microvesicles, are small membranous structures secreted by nearly all cell types. They have emerged as crucial mediators in intercellular communication, playing pivotal roles in diverse physiological and pathological processes, notably within the realm of immunity. These roles go beyond mere cellular interactions, as extracellular vesicles stand as versatile and dynamic components of immune regulation, impacting both innate and adaptive immunity. Their multifaceted involvement includes immune cell activation, antigen presen- tation, and immunomodulation, emphasising their significance in maintaining immune homeostasis and contributing to the pathogenesis of immune-related disorders. Extracellular vesicles participate in immunomodulation by delivering a wide array of bioactive molecules, including proteins, lipids, and nucleic acids, thereby influencing gene expression in target cells. This manuscript presents a comprehensive review that encompasses in vitro and in vivo studies aimed at elucidating the mechanisms through which EVs modulate human immunity. Understanding the intricate interplay between extracellular vesicles and immunity is imperative for unveiling novel therapeutic targets and diagnostic tools applicable to various immunological disorders, including autoimmune diseases, infectious diseases, and cancer. Furthermore, recognising the potential of EVs as versatile drug delivery vehicles holds significant promise for the future of immunotherapies.

Mitochondrial DNA (mtDNA) is a 16,569 base pairs, double-stranded, circular molecule that contains 37 genes coding for 13 subunits of the respiratory chain plus 2 rRNAs and 22 tRNAs. Mutations in these genes have been identified in patients with a variety of disorders affecting every system in the body. The advent of next generation sequencing technologies has provided the possibility to perform the whole mitochondrial DNA sequencing, allowing the identification of disease-causing pathogenic variants in a single platform. In this study, the whole mtDNA of 100 patients from South Italy affected by mitochondrial diseases was analyzed by using an amplicon-based approach and then the enriched libraries were deeply sequenced on the Ion Torrent platform from Thermofisher Scientific. After bioinformatics analysis, we were able to find 26 nonsynonymous variants with a MAF <1% that were associated with different pathological phenotypes, expanding the mutational spectrum of these diseases. Moreover, among the new mutations found, we have also analyzed the 3D structure of the MT-ATP6 A200T gene variation in order to confirm suspected functional alterations. This work brings light on new variants possibly associated with several mitochondriopathies in patients from South Italy and confirms that deep sequencing approach, compared to the standard methods, is a reliable and time-cost reducing strategy to detect all the variants present in the mitogenome, making the possibility to create a genomics landscape of mitochondrial DNA variations in human diseases.

The flavoenzyme N-ribosyldihydronicotinamide (NRH):quinone oxidoreductase 2 (NQO2) catalyzes two-electron reductions of quinones. NQO2 contributes to the metabolism of biogenic and xenobiotic quinones, including a wide range of antitumor drugs, with both toxifying and detoxifying functions. Moreover, NQO2 activity can be inhibited by several compounds, including drugs and phytochemicals such as flavonoids. NQO2 may play important roles that go beyond quinone metabolism and include the regulation of oxidative stress, inflammation, and autophagy, with implications in carcinogenesis and neurodegeneration. NQO2 is a highly polymorphic gene with several allelic variants, including insertions (I), deletions (D) and single-nucleotide (SNP) polymorphisms located mainly in the promoter, but also in other regulatory regions and exons. This is the first systematic review of the literature reporting on NQO2 gene variants as risk factors in degenerative diseases or drug adverse effects. In particular, hypomorphic 29 bp I alleles have been linked to breast and other solid cancer susceptibility as well as to interindividual variability in response to chemotherapy. On the other hand, hypermorphic polymorphisms were associated with Parkinson's and Alzheimer's disease. The I and D promoter variants and other NQO2 polymorphisms may impact cognitive decline, alcoholism and toxicity of several nervous system drugs. Future studies are required to fill several gaps in NQO2 research.

PM2.5 exposure represents a risk factor for the public health. PM2.5 is able to cross the blood-alveolar and blood-brain barriers and reach the brain through three routes: nasal olfactory pathway, nose-brain pathway, blood-brain barrier pathway. We evaluated the effect of PM2.5 to induce cytotoxicity and reduced viability on in vitro cultures of OECs (Olfactory Ensheathing Cells) and SH-SY5Y cells. PM2.5 samples were collected in the metropolitan area of Catania, and the gravimetric determination of PM2.5, characterization of 10 trace elements and 16 polycyclic aromatic hydrocarbons (PAHs) were carried out for each sample. PM2.5 extracts were exposed to cultures of OECs and SH-SY5Y cells for 24-48-72 h, and the cell viability assay (MTT) was evaluated. Assessment of mitochondrial and cytoskeleton damage, and the assessment of apoptotic process were performed in the samples that showed lower cell viability. We have found an annual average value of PM2.5 = 16.9 ?g/m3 and a maximum value of PM2.5 = 27.6 ?g/m3 during the winter season. PM2.5 samples collected during the winter season also showed higher concentrations of PAHs and trace elements. The MTT assay showed a reduction in cell viability for both OECs (44%, 62%, 64%) and SH-SY5Y cells (16%, 17%, 28%) after 24-48-72 h of PM2.5 exposure. Furthermore, samples with lower cell viability showed a decrease in mitochondrial membrane potential, increased cytotoxicity, and also impaired cellular integrity and induction of the apoptotic process after increased expression of vimentin and caspase-3 activity, respectively. These events are involved in neurodegenerative processes and could be triggered not only by the concentration and time of exposure to PM2.5, but also by the presence of trace elements and PAHs on the PM2.5 substrate. The identification of more sensitive cell lines could be the key to understanding how exposure to PM2.5 can contribute to the onset of neurodegenerative processes.

Background: Fabry's Disease (FD) is a rare, multi-organ lysosomal disease, caused by the deficiency of the enzyme ?-galactosidase A. This leads to the accumulation of glycosphingolipids, particularly globotriaosylsphingosine (Lyso-Gb3), in visceral tissues and vascular endothelium throughout the body, with renal, cardiac, and central nervous system damage such that quality and expectancy of life are impaired. Progressive accumulation of Gb3 in podocytes, epithelial cells and the tubular cells of the distal tubule and loop of Henle contribute to the renal symptoms of Fabry disease, which manifest as proteinuria and reduced glomerular filtration rate leading to chronic kidney disease and progression to endstage renal disease. We report the case of a patient with hypertension and proteinuria whose diagnosis of Fabry's disease was suspected due to the ultrasound finding of parapelvic cysts. Case presentation: A 41-year-old man with proteinuria and hypertension presented to our outpatient Department. The patient underwent a genetic investigation for Fabry due to the ultrasound finding of parapelvic cysts. We found a novel mutation c.160del in exon 1 of GLA gene with aminoacidic change in p. (Asp55ThrfsTer66). The new mutation was also found in the mother and his brother. In males, the elevated accumulation of LysoGB3 was associated with the presence of renal parapelvic cysts. Conclusion: This mutation is not reported in Fabry disease-associated mutation databases and has not been previously described in the literature but, the absent enzyme activity found in our patient, the significant blood accumulation of LysoGb3, as well as the type of mutation suggest its pathogenetic role. In addition, the novel mutation would appear to correlate with the occurrence of pelvic cysts in male subjects. Therefore Fabry's disease should be suspected in male patients with hypertension, proteinuria and ultrasonographic finding of parapelvic cysts, especially in patients with otherwise unexplained cardiac, neurologic and/or ocular abnormalities. Further studies are needed to corroborate the finding.

Background: Anderson-Fabry Disease (AFD), a rare multisystemic lysosomal storage disease, is caused by GLA gene mutation leading to ?-galactosidase A (?-Gal A) deficiency. AFD exhibits an X-linked inheritance pattern, with a more pronounced and early-onset phenotype in males. The phenotypic manifestation in females is variable due to cellular mosaicism resulting from lyonization of the X chromosome, insidious and potentially severe, particularly with regard to cardiac involvement. Objective: To assess the differences in cardiac manifestations of AFD in women (F) versus men (M), in relation to the type of mutation involved: classical (CL), late-onset [LO] or variants of uncertain significance [VUS]).

Background: La malattia di Anderson-Fabry (AFD), rara patologia da accumulo lisosomiale ad interessamento multisistemico, causata dalla mutazione del gene GLA, che codifica per l'enzima ?-galattosidasi A (?-Gal A), presenta un pattern di trasmissione ereditaria X-linked, manifestando un fenotipo più marcato ad esordio precoce nei maschi (forma classica). Nelle donne la manifestazione fenotipica è variabile ed insidiosa a causa del mosaicismo cellulare derivante dalla lyonizzazione del cromosoma X, che determina una presentazione della malattia più sfumata, coinvolgente talvolta singoli organi, ma imprevedibile e potenzialmente grave, in particolar modo a livello cardiaco. Obiettivo: Valutare le differenze nella manifestazione cardiologica della AFD nelle donne (F) rispetto agli uomini (M), esaminando e confrontando anche il tipo di mutazione coinvolta (classica, late-onset [LO] o variante a significato incerto [VUS]).

ntroduction: Long non-coding RNA H19 (lncH19) is highly expressed in colorectal cancer (CRC) and plays critical roles in tumor development, proliferation, metastasis, and drug resistance. Indeed, the expression of lncH19 usually affects the outcomes of chemo-, endocrine, and targeted therapies. ITF2357 (givinostat) is a histone deacetylase inhibitor (HDACi) that revealed a significant anti-tumor action by inducing apoptosis in different tumor models, including leukemia, melanoma, and glioblastoma. However, no data are present in the literature regarding the use of this compound for CRC treatment. Here, we investigate the role of lncH19 in ITF2357-induced apoptosis in CRC cells. Methods: The HCT-116 CRC cell line was stably silenced for H19 to investigate the role of this lncRNA in ITF2357-induced cell death. Cell viability assays and flow cytometric analyses were performed to assess the anti-proliferative and pro- apoptotic effects of ITF2357 in CRC cell lines that are silenced or not for lncH19. RT-PCR and Western blot were used to study the effects of ITF2357 on autophagy and apoptosis markers. Finally, bioinformatics analyses were used to identify miRNAs targeting pro-apoptotic factors that can be sponged by lncH19. Results: ITF2357 increased the expression levels of H19 and reduced HCT-116 cell viability, inducing apoptosis, as demonstrated by the increase in annexin-V positivity, caspase 3 cleavage, and poly (ADP-ribose) polymerase (PARP-1) degradation. Interestingly, the apoptotic effect of ITF2357 was much less evident in lncH19-silenced cells. We showed that lncH19 plays a functional role in the pro-apoptotic activity of the drug by stabilizing TP53 and its transcriptional targets, NOXA and PUMA. ITF2357 also induced autophagy in CRC cells, which was interpreted as a pro-survival response not correlated with lncH19 expression. Furthermore, ITF2357 induced apoptosis in 5- fluorouracil-resistant HCT-116 cells that express high levels of lncH19. Conclusion: This study shows that lncH19 expression contributes to ITF2357- induced apoptosis by stabilizing TP53. Overall, we suggest that lncH19 expression may be exploited to favor HDACi-induced cell death and overcome 5-fluorouracil chemoresistance.

In the last years, extracellular vesicles (EVs) from different plant matrices have been isolated and gained the interest of the scientific community for their intriguing biological properties. In this study, we isolated and characterized nano- vesicles from lemon juice (LNVs) and evaluated their antioxidant effects. We tested LNV antioxidant activity using human dermal fibroblasts that were pre- treated with LNVs for 24 h and then stimulated with hydrogen peroxide (H2O2) and UVB irradiation. We found that LNV pre-treatment reduced ROS levels in fi- broblasts stimulated with H2O2 and UVB. This reduction was associated with the activation of the AhR/Nrf2 signaling pathway, whose protein expression and nu- clear localization was increased in fibroblasts treated with LNVs. By using zebra- fish embryos as in vivo model, we confirmed the antioxidant effects of LNVs. We found that LNVs reduced ROS levels and neutrophil migration in zebrafish em- bryos stimulated with LPS.

Background Metastatic disease is the major cause of cancer-related deaths. Increasing evidence shows that pri- mary tumor cells can promote metastasis by preparing the local microenvironment of distant organs, inducing the formation of the so-called "pre-metastatic niche". In recent years, several studies have highlighted that among the tumor-derived molecular components active in pre-metastatic niche formation, small extracellular vesicles (sEVs) play a crucial role. Regarding liver metastasis, the ability of tumor-derived sEVs to affect the activities of non-parenchymal cells such as Kupffer cells and hepatic stellate cells is well described, while the effects on hepatocytes, the most con- spicuous and functionally relevant hepatic cellular component, remain unknown. Methods sEVs isolated from SW480 and SW620 CRC cells and from clinical samples of CRC patients and healthy sub- jects were used to treat human healthy hepatocytes (THLE-2 cells). RT-qPCR, Western blot and confocal microscopy were applied to investigate the effects of this treatment. Results Our study shows for the first time that TGF?1-carrying CRC_sEVs impair the morphological and func- tional properties of healthy human hepatocytes by triggering their TGF?1/SMAD-dependent EMT. These abilities of CRC_sEVs were further confirmed by evaluating the effects elicited on hepatocytes by sEVs isolated from plasma and biopsies from CRC patients. Conclusions Since it is known that EMT of hepatocytes leads to the formation of a fibrotic environment, a well- known driver of metastasis, these results suggest that CRC_sEV-educated hepatocytes could have an active and until now neglected role during liver metastasis formation.

La Distrofia dei Cingoli Recessiva di tipo 8 (LGMDR8) è stata descritta per la prima volta nelle popolazioni Hutterite del Nord America associata alla mutazione D487N localizzata nel gene che codifica TRIM32. Tale proteina appartiene ad una grande famiglia caratterizzata da un motivo tripartito, coinvolta in processi di ubiquitinazione con attività E3 ligasica.TRIM32 possiede a partire dalla porzione N-terminale un dominio RING, un B-box, un dominio Coiled-Coil e nella porzione C-terminale sei regioni ripetute NHL essenziali per l'interazione della proteina con i suoi substrati. Noi descriviamo un paziente di 54 anni del sud Italia affetto da distrofia dei cingoli. Già nella prima decade di vita, il paziente ha mostrato difficoltà nella deambulazione e stazione eretta peggiorati nel corso degli anni, ipotrofia dei cingoli specie scapolari con deficit del deltoide e bicipite laterale. Agli arti inferiori il deficit è accentuato sui tibiali anteriori e peronieri. Il paziente mostrava una moderata iperCkemia, segno di Gowers positivo, ROT ridotti ai 4 arti, insufficienza ventilatoria. L'analisi immunoistologica del tessuto muscolare bioptico del deltoide ha mostrato fibre ipo/ipertrofiche di forma prevalentemente rotondeggianti, diverse fibre con nuclei in posizione centrale, aumento di collagene, tessuto adiposo e della quota lipidica. L'analisi molecolare condotta mediante NGS sul DNA del paziente ha permesso l'identificazione della variante R596G in omozigosi nel gene TRIM32. Questa variante è già stata riportata in letteratura come probabilmente patologica correlata a LGMDR8. R596G ricade nel sesto dominio NHL del gene, critico per il riconoscimento delle proteine target che devono essere ubiquitinilate. Tutte le analisi in silico come Polyphen, MutPred2, Polyphen2, Panther, Sift, Mutation Taster sono a supporto di un suo effetto deleterio. Il paziente presentava, inoltre, la mutazione Leu276Ile in eterozigosi nel gene FKRP che è la mutazione più frequentemente associata a LGMDR9. La presenza di entrambe le varianti nel gene FKRP E TRIM32 potrebbero avere un effetto additivo sul fenotipo del paziente. Entrambe le proteine sono essenziali per mantenere l'integrità del costamero e dunque l'efficienza della contrazione muscolare.

TRIM32 is a member of the Tripartite-Motif family of proteins characterized by a RING finger, a B-box motif, a coiled-coil region and six NHL-repeats. TRIM32 has E3 ubiquitin ligase activity by participating in myofibrillar protein turnover. D487N mutation in the TRIM32 gene, has been associated with Limb Girdle Muscular Dystrophy Recessive type 8 (LGMDR8) in the inbred Hutterite population. We reported R596G homozygous missense variant in TRIM32 gene in a 54-year-old patient with LGMD from south Italy.

Abstract Background: Gaucher disease (GD) is a rare autosomal recessive inherited disorder caused by the lysosomal enzyme acid ?-glucosidase deficiency. Many patients experience a critical delay in the diagnosis of up to 8-10 years due to its rarity and variability in signs and symptoms, with the consultation of several specialists. Patients and methods: This prospective observational study analyzed the prevalence of GD in 600 patients with monoclonal gammopathy of uncertain significance (MGUS) from January 2018 until February 2022. Results: The mean age of participants was 66 years, with a mean monoclonal component of 0.58 g/dL. In 433 MGUS patients with available data, anemia (hemoglobin level < 10 g/dL) was present in 31 patients (7%), and thrombocytopenia (platelet count <100.000/mm3 ) in 24 (5.5%). Of 600 MGUS patients tested for acid ?-glucosidase enzyme activity, 7 patients (1.2%) had activity below 2.5 nmol/h/mL. In comparison, GBA gene analysis was executed in 110 patients. It revealed 4 patients (0.7%) affected by GD (3 patients with compound heterozygous mutation and 1 with homozygous mutation), with a prevalence of 1 every 150 MGUS patients. Furthermore, 12 out of the remaining 106 evaluated patients (11%) were carriers of a single heterozygous mutation while having regular enzyme activity. Conclusions: The clinical heterogeneity of GD and frequent lack of awareness among physicians often lead to diagnostic delays and severe clinical manifestations. The role of MGUS in the presence of at least one clinical sign, such as low platelet count, organomegaly, bone pain, or bleeding tendency, could aid in initiating GD screening with DBS, thus reducing the period between symptom onset and the diagnosis of this rare disease.

Sea urchins are emblematic models in developmental biology and display several characteristics that set them apart from other deuterostomes. To uncover the genomic cues that may underlie these specificities, we generated a chromosome-scale genome assembly for the sea urchin Paracentrotus lividus and an extensive gene expression and epigenetic profiles of its embryonic development. We found that, unlike vertebrates, sea urchins retained ancestral chromosomal linkages but underwent very fast intrachromosomal gene order mixing. We identified a burst of gene duplication in the echinoid lineage and showed that some of these expanded genes have been recruited in novel structures (water vascular system, Aristotle's lantern, and skeletogenic micromere lineage). Finally, we identified gene-regulatory modules conserved between sea urchins and chordates. Our results suggest that gene-regulatory networks controlling development can be conserved despite extensive gene order rearrangement.

In respiratory infections, anemia is both a consequence of acute inflammation and a predictor of poor clinical outcomes. There are few studies investigating the role of anemia in COVID-19, suggesting a potential role in predicting disease severity. In this study, we aimed to assess the association between the presence of anemia at admission and incidence of severe disease and death in patients hospitalized for COVID-19. Data from all adult patients admitted for COVID-19 in University Hospital "P. Giaccone" Palermo, and University Hospital of Bari, Italy, were retrospectively collected from 1st of September 2020 to 31 August 2022. The association between anemia (defined as Hb < 13 g/dl and < 12 g/dl in males and females, respectively), in-hospital mortality and severe COVID-19 was tested using a Cox's regression analysis. Severe COVID-19 forms were defined as admission to intensive or sub-intensive care unit or a qSOFAscore >= 2 or CURB65scores >= 3. p values were calculated using the Student's t test for continuous variables and the Mantel-Haenszel Chi-square test for categorical ones. The association between anemia and the mortality was made using a Cox's regression analysis, adjusted, in two models, for the potential confounders and using a propensity score. Among the 1562 patients included in the analysis, prevalence of anemia was 45.1% (95% CI 43-48%). Patients with anemia were significantly older (p < 0.0001), reported more co-morbidities, and presented higher baseline levels of procalcitonin, CRP, ferritin and IL-6. Overall, the crude incidence of mortality was about four times higher in patients with anemia compared to those without. After adjusting for 17 potential confounders, the presence of anemia significantly increased the risk of death (HR = 2.68; 95% CI: 1.59-4.52) and of risk of severe COVID-19 (OR = 2.31; 95% CI: 1.65-3.24). The propensity score analysis substantially confirmed these analyses. Our study provides evidence that, in patients hospitalized for COVID-19, anemia is both associated with a more pronounced baseline pro-inflammatory profile and higher incidence of in-hospital mortality and severe disease.

Alzheimer's disease (AD) is characterized by agglomerated proteins constitued by amyloid-beta (A?). A? possesses neutoxic effect and is also a substrate of tissue transglutaminase (TG2), an ubiquitarian protein that plays a key role in AD. Several reports indicated that in AD is involved oxidative stress and that the treatment with antioxidants mitigates the effects of oxidative stress in the central nervous system. In particular, astaxanthin, an antioxidant with antiflammatory properties, could have an important therapeutic role in AD. Herein, we have evaluated the effect of astaxanthin pretreatment on olfactory ensheathing cells (OECs) exposed to the native peptide of A?(1-42) or its fragments A?(25-35) and A?(35-25). OECs are glial cells located in the olfactory system, the first to show a deficit in neurodegenerative diseases. Vimentin, GFAP, nestin, cyclin D1, TG2 expression and the activation of the apoptotic pathway were assessed through immunocytochemical techniques. The percentage of cell viability and ROS levels were detected. In addition, to monitor the mitochondrial status, we used delayed luminescence (DL). We found that astaxanthin was able to reduce TG2 expression up-regulated by A?, reducing also GFAP and Vimentin levels. Astaxanthin pre-treatment stimulates cellular repair increasing cyclin D1 and nestin levels and inhibing apoptotic pathway activation. In parallel, we observed a significant change of DL intensity in OECs exposed to the toxic fragment A? (25-35), that completely disappear when OECs were pre-incubated with astaxantin. Therefore, we can suggest that astaxanthin pretreatment could represent an innovative mechanism to counteract the aberrant TG2 overexpression in AD.

In recent years, research has focused on colorectal cancer to implement modern treatment approaches to improve patient survival. In this new era, ?? T cells constitute a new and promising candidate to treat many types of cancer because of their potent killing activity and their ability to recognize tumor antigens independently of HLA molecules. Here, we focus on the roles that ?? T cells play in antitumor immunity, especially in colorectal cancer. Furthermore, we provide an overview of small-scale clinical trials in patients with colorectal cancer employing either in vivo activation or adoptive transfer of ex vivo expanded ?? T cells and suggest possible combinatorial approaches to treat colon cancer.