Expression and localization of CHI3L1 in monocyte derived dendritic cells

Chitinase-3-like-1 protein (CHI3L1) also called YKL-40, is a 40 kDa mammalian glycoprotein which is a heparin, chitin and collagen binding member of the mammalian chitinase-like proteins. Biological activities of CHI3L1 embrace regulation of cell proliferation, adhesion, angiogenesis, migration and activation. CHI3L1 is produced by variety of cells, including neutrophils, monocytes/macrophages, osteoclasts and Kupffer cells [1]. CHI3L1 secretion is induced by interferon (INF)-g and interleukin (IL)-6 and is an acute phase reactant associated with disease severity and mortality in a variety of infectious [2]. In this study, we have examined the expression and localization of CHI3L1 during the differentiation and maturation of monocyte derived dendritic cells by real time RT-PCR, Western Blot, Confocal Immunofluorescence, and Immunocytochemical assays. Potential nuclear localization signal (NLS) was determinated using the open source software cNLS Mapper and Chimera. Peripheral blood monocytes were differentiated toward immature DCs (iDC) and mature DCs (mDCs) through a combination of factors and cytokines. Our result showed, for the first time, that CHI3L1 is expressed during the process of differentiation and maturation of DCs in time dependent manner. Furthermore, CHI3L1 is evenly distributed in cytoplasm and in the nucleus of both the iDCs and mDCs. In conclusion, the discovery of CHI3L1 expression in DCs has opened new dilemma for designing DC-based cancer immunotherapeutic. In fact, on the light of these results one can’t exclude that as well as activated Tumor-associated macrophages (TAMs) also DCs infiltration could to be a significant unfavorable prognostic factor for cancer patients

Chitotriosidase expression in monocyte-derived dendritic cells

Chitotriosidase (CHIT1) belongs to 18 glycosyl-hydrolase family, an ancient gene family that is widely expressed from prokaryotes to eukaryotes [1]. CHIT1 is a very critical enzyme to regulate the susceptibility to infection of organisms containing chitin as structural components. Conversely, during the development of acute/chronic inflammatory disorders, the enzymatic activity of CHIT1 increases significantly. The CHIT1 is expressed in activated macrophages as well in different lines monocyte-derived such as Kupffer cells and osteoclasts [2]. So far, it is unknown whether CHIT1 is expressed in other cells involved in the immune response such as monocyte-derived DCs. In this study we have investigated whether CHIT1 is produced in monocyte-derived DCs (moDCs) and the differential expression of CHIT1 during the different stage of moDCs differentiation. The presence of CHIT1 were examined by real time RT-PCR, Western Blot and Confocal Immunofluorescence, in Immature Dendritic cells (iDCs), generated from human monocytes by stimulation with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) and in mature Dendritic cells (mDCs), obtained by using lipopolysaccharide (LPS) and interferon-gamma (IFN-g). We observed that CHIT1 was expressed during the DCs differentiation and maturation process in time dependent manner. The maturation of DCs showed a significantly increased expression of CHIT1 mRNA and protein. Furthermore, the CHIT1 was evenly distributed in cytoplasm both in iDCs and in mDCs. The enzymatic activity confirmed that CHIT1 could play a role in moDCs function. Taken together, our data confirm the crucial role of CHIT1 in primary immune responses and indicate that could be correlated with the immunogenicity of DCs

Clinical Features, Cardiovascular Risk Profile, and Therapeutic Trajectories of Patients with Type 2 Diabetes Candidate for Oral Semaglutide Therapy in the Italian Specialist Care

Introduction: This study aimed to address therapeutic inertia in the management of type 2 diabetes (T2D) by investigating the potential of early treatment with oral semaglutide. Methods: A cross-sectional survey was conducted between October 2021 and April 2022 among specialists treating individuals with T2D. A scientific committee designed a data collection form covering demographics, cardiovascular risk, glucose control metrics, ongoing therapies, and physician judgments on treatment appropriateness. Participants completed anonymous patient questionnaires reflecting routine clinical encounters. The preferred therapeutic regimen for each patient was also identified. Results: The analysis was conducted on 4449 patients initiating oral semaglutide. The population had a relatively short disease duration (42%  60% of patients, and more often than sitagliptin or empagliflozin. Conclusion: The study supports the potential of early implementation of oral semaglutide as a strategy to overcome therapeutic inertia and enhance T2D management

Acute Delta Hepatitis in Italy spanning three decades (1991–2019): Evidence for the effectiveness of the hepatitis B vaccination campaign

Updated incidence data of acute Delta virus hepatitis (HDV) are lacking worldwide. Our aim was to evaluate incidence of and risk factors for acute HDV in Italy after the introduction of the compulsory vaccination against hepatitis B virus (HBV) in 1991. Data were obtained from the National Surveillance System of acute viral hepatitis (SEIEVA). Independent predictors of HDV were assessed by logistic-regression analysis. The incidence of acute HDV per 1-million population declined from 3.2 cases in 1987 to 0.04 in 2019, parallel to that of acute HBV per 100,000 from 10.0 to 0.39 cases during the same period. The median age of cases increased from 27 years in the decade 1991-1999 to 44 years in the decade 2010-2019 (p < .001). Over the same period, the male/female ratio decreased from 3.8 to 2.1, the proportion of coinfections increased from 55% to 75% (p = .003) and that of HBsAg positive acute hepatitis tested for by IgM anti-HDV linearly decreased from 50.1% to 34.1% (p < .001). People born abroad accounted for 24.6% of cases in 2004-2010 and 32.1% in 2011-2019. In the period 2010-2019, risky sexual behaviour (O.R. 4.2; 95%CI: 1.4-12.8) was the sole independent predictor of acute HDV; conversely intravenous drug use was no longer associated (O.R. 1.25; 95%CI: 0.15-10.22) with this. In conclusion, HBV vaccination was an effective measure to control acute HDV. Intravenous drug use is no longer an efficient mode of HDV spread. Testing for IgM-anti HDV is a grey area requiring alert. Acute HDV in foreigners should be monitored in the years to come

Tinnitus and organ preservation protocols in head and neck cancers treatment: a descriptive review of the literature

INTRODUCTION: Head and neck cancer represents the sixth most common cancer group worldwide and significant progress has been made in the non-surgical management. however, the possibility of audiological side effects has increased. in this article we performed a review of the literature to analyze the principal ear side effects derived from non-surgical protocols. EViDENCE aCQUiSitiON: We performed a review of the international literature. For each eligible article, we collected the study design, the sample size and the demographical characteristics, the organ preservation protocols, and the findings about tinnitus or hearing loss. Due to the lack of homogeneity of the data extracted we performed a descriptive review of the literature. EViDENCE SyNthESiS: initial search found 350 studies and 332 were excluded; 18 were reviewed in full text; 10 studies met the inclusion criteria. Overall, 3606 patients were included. the number of patients ranged between 43 and 880. the age ranged between 9 and 84 years. the males are 1878 and females are 931. In 796 case the sex was not specified. Seven studies are monocentric and retrospective; two studies are multicentric and retrospective and only one study is prospective, randomized and observational with the patients randomized in three groups. Data on tinnitus or hearing impairment are evaluated by questionaries that analyze the patient-reported outcomes or by auditory evaluation like the pure tone audiometry or the auditory Brain recordings. CONClUSiONS: the management of audiological side effects is fundamental in order to guarantee a good quality of life. and rehabilitation should be started immediately after the therapies. all ENt specialist, head and neck surgeons, oncologists and radiotherapist oncologists must be vigilant about ototoxicity and implement the strategies to decrease additional morbidity due to ototoxicity

Studies on the Accumulation of Secondary Metabolites and Evaluation of Biological Activity of In Vitro Cultures of <i>Ruta montana</i> L. in Temporary Immersion Bioreactors

The present work focuses on in vitro cultures of Ruta montana L. in temporary immersion PlantformTM bioreactors. The main aim of the study was to evaluate the effects of cultivation time (5 and 6 weeks) and different concentrations (0.1–1.0 mg/L) of plant growth and development regulators (NAA and BAP) on the increase in biomass and the accumulation of secondary metabolites. Consequently, the antioxidant, antibacterial, and antibiofilm potentials of methanol extracts obtained from the in vitro-cultured biomass of R. montana were evaluated. High-performance liquid chromatography analysis was performed to characterize furanocoumarins, furoquinoline alkaloids, phenolic acids, and catechins. The major secondary metabolites in R. montana cultures were coumarins (maximum total content of 1824.3 mg/100 g DM), and the dominant compounds among them were xanthotoxin and bergapten. The maximum content of alkaloids was 561.7 mg/100 g DM. Concerning the antioxidant activity, the extract obtained from the biomass grown on the 0.1/0.1 LS medium variant, with an IC50 0.90 ± 0.03 mg/mL, showed the best chelating ability among the extracts, while the 0.1/0.1 and 0.5/1.0 LS media variants showed the best antibacterial (MIC range 125–500 µg/mL) and antibiofilm activity against resistant Staphylococcus aureus strains

Cytostatic Effects of Polyethyleneimine Surfaces on the Mesenchymal Stromal Cell Cycle

Polyelectrolytes assembled layer-by-layer (PEMs) are commonly used as functional coatings to build-up biological interfaces, particularly suitable as compatible layers for the interaction with a biological medium, providing suitable conditions to promote or prevent cell seeding while maintaining the phenotype. The proper assessment of the biocompatibility of PEMs and the elucidation of the related mechanisms are therefore of paramount importance. In this study, we report in detail the effect of two different PEM endings, polystyrene sulfonate (PSS) and polyethylenimine (PEI), respectively, on the cell adhesion, growth, and viability of human bone mesenchymal stromal cells (MSCs). The results have shown that PSS-ended substrates appear to be the most suitable to drive the cell adhesion and phenotype maintenance of MSCs, showing good biocompatibility. On the contrary, while the cells seem to adhere more quickly and strongly on the PEI-ended surfaces, the interaction with PEI significantly affects the growth and viability, reducing the cell spreading capability, by sequestering the adhesion molecules already in the very early steps of cell&ndash;substrate contact. These results point to the promotion of a cytostatic effect of PEI, rather than the often-claimed cytotoxicity

Synovial Fluid-Derived Extracellular Vesicles of Patients with Arthritides Contribute to Hippocampal Synaptic Dysfunctions and Increase with Mood Disorders Severity in Humans

Arthritides are a highly heterogeneous group of disorders that include two major clinical entities, localized joint disorders such as osteoarthritis (OA) and systemic autoimmune-driven diseases such as rheumatoid arthritis (RA). Arthritides are characterized by chronic debilitating musculoskeletal conditions and systemic chronic inflammation. Poor mental health is also one of the most common comorbidities of arthritides. Depressive symptoms which are most prevalent, negatively impact patient global assessment diminishing the probability of achieving the target of clinical remission. Here, we investigated new insights into mechanisms that link different joint disorders to poor mental health, and to this issue, we explored the action of the synovial fluid-derived extracellular vesicles (EVs) on neuronal function. Our data show that the exposure of neurons to different concentrations of EVs derived from both RA and OA synovial fluids (RA-EVs and OA-EVs) leads to increased excitatory synaptic transmission but acts on specific modifications on excitatory or inhibitory synapses, as evidenced by electrophysiological and confocal experiments carried out in hippocampal cultures. The treatment of neurons with EVs membrane is also responsible for generating similar effects to those found with intact EVs suggesting that changes in neuronal ability arise upon EVs membrane molecules&prime; interactions with neurons. In humans with arthritides, we found that nearly half of patients (37.5%) showed clinically significant psychiatric symptoms (CGIs score &ge; 3), and at least mild anxiety (HAM-A &ge; 7) or depression (MADRS and HAM-D &ge; 7); interestingly, these individuals revealed an increased concentration of synovial EVs. In conclusion, our data showing opposite changes at the excitatory and inhibitory levels in neurons treated with OA- and RA-EVs, lay the scientific basis for personalized medicine in OA and RA patients, and identify EVs as new potential actionable biomarkers in patients with OA/RA with poor mental health

IGFBP-6 Alters Mesenchymal Stromal Cell Phenotype Driving Dasatinib Resistance in Chronic Myeloid Leukemia

Chronic myeloid leukemia (CML), BCR-ABL1-positive, is classified as a myeloproliferative characterized by Philadelphia chromosome/translocation t(9;22) and proliferating granulocytes. Despite the clinical success of tyrosine kinase inhibitors (TKi) agents in the treatment of CML, most patients have minimal residual disease contained in the bone marrow microenvironment, within which stromal cells assume a pro-inflammatory phenotype that determines their transformation in cancer-associated fibroblasts (CAF) which, in turn can play a fundamental role in resistance to therapy. Insulin-like Growth Factor Binding Protein-6 (IGFBP-6) is expressed during tumor development, and is involved in immune-escape and inflammation as well, providing a potential additional target for CML therapy. Here, we aimed at investigating the role of IGFBP-6/SHH/TLR4 axis in TKi response. We used a CML cell line, LAMA84-s, and healthy bone marrow stromal cells, HS-5, in mono- or co-culture. The two cell lines were treated with Dasatinib and/or IGFBP-6, and the expression of inflammatory markers was tested by qRT-PCR; furthermore, expression of IGFBP-6, TLR4 and Gli1 were evaluated by Western blot analysis and immumocytochemistry. The results showed that both co-culture and Dasatinib exposure induce inflammation in stromal and cancer cells so that they modulate the expression of TLR4, and these effects were more marked following IGFBP-6 pre-treatment suggesting that this molecule may confer resistance through the inflammatory processes. This phenomenon was coupled with sonic hedgehog (SHH) signaling. Indeed, our data also demonstrate that HS-5 treatment with PMO (an inducer of SHH) induces significant modulation of TLR4 and overexpression of IGFPB-6 suggesting that the two pathways are interconnected with each other and with the TLR-4 pathway. Finally, we demonstrated that pretreatment with IGFBP-6 and/or PMO restored LAMA-84 cell viability after treatment with Dasatinib, suggesting that both IGFBP-6 and SHH are involved in the resistance mechanisms induced by the modulation of TLR-4, thus indicating that the two pathways may be considered as potential therapeutic targets

New Data on Native and Alien Vascular Flora of Sicily (Italy): New Findings and Updates

In this paper, based on fieldwork and herbaria surveys, new data concerning the presence of 32 native and alien vascular species for Sicily (Italy) are provided. Among the native species, the occurrence of the following taxa is reported for the first time or confirmed after many decades of non-observation: Aira multiculmis, Arum maculatum, Carex flacca subsp. flacca, Mentha longifolia, Oxybasis chenopodioides, Najas minor and Xiphion junceum. Furthermore, we document the presence of three native species (Cornus mas, Juncus foliosus and Limonium avei) that, despite being repeatedly observed in Sicily and reported in the literature, are inexplicably omitted by the most recent authoritative checklists regarding the flora of Italy. Finally, fifteen alien species new to Sicily (including one new to Europe, i.e., Pyrus betulifolia) are reported and seven poorly documented allochthonous taxa are confirmed for the island, and for two of them, a status change is proposed. These new or confirmed records allow us to better define the European and national distribution of the targeted taxa and offer new insights on the native and alien flora of Sicily