Human γδ T-Cells: from surface receptors to the therapy of high-risk leukemias

γδ T lymphocytes are potent effector cells, capable of efficiently killing tumor and leukemia cells. Their activation is mediated by γδ T-cell receptor (TCR) and by activating receptors shared with NK cells (e.g., NKG2D and DNAM-1). γδ T-cell triggering occurs upon interaction with specific ligands, including phosphoantigens (for Vγ9Vδ2 TCR), MICA-B and UL16 binding protein (for NKG2D), and PVR and Nectin-2 (for DNAM-1). They also respond to cytokines undergoing proliferation and release of cytokines/chemokines. Although at the genomic level γδ T-cells have the potential of an extraordinary TCR diversification, in tissues they display a restricted repertoire. Recent studies have identified various γδ TCR rearrangements following either hematopoietic stem cell transplantation (HSCT) or cytomegalovirus infection, accounting for their "adaptive" potential. In humans, peripheral blood γδ T-cells are primarily composed of Vγ9Vδ2 chains, while a minor proportion express Vδ1. They do not recognize antigens in the context of MHC molecules, thus bypassing tumor escape based on MHC class I downregulation. In view of their potent antileukemia activity and absence of any relevant graft-versus-host disease-inducing effect, γδ T-cells may play an important role in the successful clinical outcome of patients undergoing HLA-haploidentical HSCT depleted of TCR αβ T/CD19+ B lymphocytes to cure high-risk acute leukemias. In this setting, high numbers of both γδ T-cells (Vδ1 and Vδ2) and NK cells are infused together with CD34+ HSC and may contribute to rapid control of infections and leukemia relapse. Notably, zoledronic acid potentiates the cytolytic activity of γδ T-cells in vitro and its infusion in patients strongly promotes γδ T-cell differentiation and cytolytic activity; thus, treatment with this agent may contribute to further improve the patient clinical outcome after HLA-haploidentical HSCT depleted of TCR αβ T/CD19+ B lymphocytes

Antimicrobial and Antibiofilm Peptides

The increasing onset of multidrug-resistant bacteria has propelled microbiology research towards antimicrobial peptides as new possible antibiotics from natural sources. Antimicrobial peptides are short peptides endowed with a broad range of activity against both Gram-positive and Gram-negative bacteria and are less prone to trigger resistance. Besides their activity against planktonic bacteria, many antimicrobial peptides also show antibiofilm activity. Biofilms are ubiquitous in nature, having the ability to adhere to virtually any surface, either biotic or abiotic, including medical devices, causing chronic infections that are difficult to eradicate. The biofilm matrix protects bacteria from hostile environments, thus contributing to the bacterial resistance to antimicrobial agents. Biofilms are very difficult to treat, with options restricted to the use of large doses of antibiotics or the removal of the infected device. Antimicrobial peptides could represent good candidates to develop new antibiofilm drugs as they can act at different stages of biofilm formation, on disparate molecular targets and with various mechanisms of action. These include inhibition of biofilm formation and adhesion, downregulation of quorum sensing factors, and disruption of the pre-formed biofilm. This review focuses on the proprieties of antimicrobial and antibiofilm peptides, with a particular emphasis on their mechanism of action, reporting several examples of peptides that over time have been shown to have activity against biofilm

NK cells and other innate lymphoid cells in hematopoietic stem cell transplantation

Natural killer (NK) cells play a major role in the T-cell depleted haploidentical hematopoietic stem cell transplantation (haplo-HSCT) to cure high-risk leukemias. NK cells belong to the expanding family of innate lymphoid cells (ILCs). At variance with NK cells, the other ILC populations (ILC1/2/3) are non-cytolytic, while they secrete different patterns of cytokines. ILCs provide host defenses against viruses, bacteria, and parasites, drive lymphoid organogenesis, and contribute to tissue remodeling. In haplo-HSCT patients, the extensive T-cell depletion is required to prevent graft-versus-host disease (GvHD) but increases risks of developing a wide range of life-threatening infections. However, these patients may rely on innate defenses that are reconstituted more rapidly than the adaptive ones. In this context, ILCs may represent important players in the early phases following transplantation. They may contribute to tissue homeostasis/remodeling and lymphoid tissue reconstitution. While the reconstitution of NK cell repertoire and its role in haplo-HSCT have been largely investigated, little information is available on ILCs. Of note, CD34+ cells isolated from different sources of HSC may differentiate in vitro toward various ILC subsets. Moreover, cytokines released from leukemia blasts (e.g., IL-1β) may alter the proportions of NK cells and ILC3, suggesting the possibility that leukemia may skew the ILC repertoire. Further studies are required to define the timing of ILC development and their potential protective role after HSCT

Tracheocutaneous fistula in patients undergoing supracricoid partial laryngectomy: the role of chronic aspiration

The aim of the present retrospective controlled study was to analyse and compare risk factors for tracheocutaneous fistula in patients who received tracheostomy after supracricoid partial laryngectomy with those who received tracheostomy for other causes. We enrolled 39 patients with tracheocutaneous fistulas who were divided into two groups. The first received temporary tracheostomy for supracricoid partial laryngectomies (n = 21), while the control group consisted of patients who received temporary tracheostomy for other causes (n = 18). Risk factors believed to play a role in the pathogenesis of tracheocutaneous fistula were examined including advanced age, cardiopathy, local infections, radiotherapy, elevated body mass index, malnutrition, decannulation time and aspiration grade. The Leipzig and Pearson scale score was significantly higher in the supracricoid partial laryngectomy group (p = 0.006 and 0.031 for univariate and multivariate analyses, respectively). The penetration/aspiration scale score was significantly higher in the supracricoid partial laryngectomy group as determined by univariate analysis (p = 0.014). The decannulation time was significantly lower in the supracricoid partial laryngectomy group (p = 0.004 and 0.0004 for univariate and multivariate analyses, respectively). The number of surgical closures for tracheocutaneous fistula was significantly higher in the supracricoid partial laryngectomy group by univariate analysis (p = 0.027). These results suggest that chronic aspiration and related cough may be important pathogenic factors for tracheocutaneous fistula and could be responsible for the significantly higher rates of closure failure in patients after supracricoid partial laryngectomy

Characterization of the proteins involved in the DNA repair mechanism in M. smegmatis.

: Several alkylating agents that either occur in the environment or are self-produced can cause DNA-damaging injuries in bacterial cells. Therefore, all microorganisms have developed repair systems that are able to counteract DNA alkylation damage. The adaptive response to alkylation stress in Escherichia coli consists of the Ada operon, which has been widely described; however, the homologous system in Mycobacterium tuberculosis (MTB) has been shown to have a different genetic organization but it is still largely unknown. In order to describe the defense system of MTB, we first investigated the proteins involved in the repair mechanism in the homologous non-pathogenic mycobacterium M. smegmatis. Ogt, Ada-AlkA and FadE8 proteins were recombinantly produced, purified and characterized. The biological role of Ogt was examined using proteomic experiments to identify its protein partners in vivo under stress conditions. Our results suggested the formation of a functional complex between Ogt and Ada-AlkA, which was confirmed both in silico by docking calculations and by gel filtration chromatography. We propose that this stable association allows the complex to fulfill the biological roles exerted by Ada in the homologous E. coli system. Finally, FadE8 was demonstrated to be structurally and functionally related to its E. coli homologous, Aid

The impact of electronic word-of-mouth management in hotel ecosystem: insights about managers'' decision-making process

Purpose There is a lack of research proving how electronic word-of-mouth (eWOM) is a valuable source of information in the hospitality industry for developing hotels'' intellectual capital. To fill this gap, this study aims to examine hotel managers'' decision-making processes regarding the acceptance and management of eWOM and its impact on the Italian hotel ecosystem. Design/methodology/approach This work takes advantage of the previous contributions to present a hotel''s decision-making process model regarding structural capital. It includes eWOM as a context variable and changes implemented as a dependent variable in a comprehensive model. The structural equation modelling applies to a database obtained through a survey addressed to Italian hotel managers. Findings The results show that eWOM plays an essential role in managers'' motivations to explain hotel changes implementation. The hotel leverages eWOM information and interaction through structural, relational and human capital to enhance products, services and strategies. Research limitations/implications This work contributes to the extant literature by providing a comprehensive framework to explain the consequences of eWOM knowledge management from the intellectual capital view in the Italian hotel ecosystem. Practical implications For practitioners, this research demonstrates how hotel managers should accept and manage eWOM knowledge through intellectual capital to make determinant decisions that improve hotel performance. Originality/value There is a scarcity of research on modelling the acceptability and management of eWOM in the hotel ecosystem from practitioners'' perspectives. This work is the first attempt to determine how eWOM knowledge management boosts hotel intellectual capital and improves service innovation and performance

Predicting FTS products through artificial neural network modelling

Fischer-Tropsch synthesis is essential for converting CO2 into hydrocarbons, creating sustainable fuels and olefins. However, challenges in production yield and reaction kinetics remain. This study introduces an artificial neural network (ANN) to predict FT synthesis products from specific inputs, including temperature, pressure, GHSV, H2/CO2 ratio, and catalyst composition (Fe weight and K as a promoter). The ANN's ability to predict outputs like CH4, C2-4, C5+, CO2 conversion, and CO selectivity, without detailed reaction mechanisms, is a key innovation. This approach circumvents complex kinetic models. The network architecture is optimized for minimal error, and results are validated against a comprehensive database

Microenvironment in neuroblastoma: Isolation and characterization of tumor-derived mesenchymal stromal cells

Background: It has been proposed that mesenchymal stromal cells (MSCs) promote tumor progression by interacting with tumor cells and other stroma cells in the complex network of the tumor microenvironment. We characterized MSCs isolated and expanded from tumor tissues of pediatric patients diagnosed with neuroblastomas (NB-MSCs) to define interactions with the tumor microenvironment. Methods: Specimens were obtained from 7 pediatric patients diagnosed with neuroblastoma (NB). Morphology, immunophenotype, differentiation capacity, proliferative growth, expression of stemness and neural differentiation markers were evaluated. Moreover, the ability of cells to modulate the immune response, i.e. inhibition of phytohemagglutinin (PHA) activated peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cytotoxic function, was examined. Gene expression profiles, known to be related to tumor cell stemness, Wnt pathway activation, epithelial-mesenchymal transition (EMT) and tumor metastasis were also evaluated. Healthy donor bone marrow-derived MSCs (BM-MSC) were employed as controls. Results: NB-MSCs presented the typical MSC morphology and phenotype. They showed a proliferative capacity superimposable to BM-MSCs. Stemness marker expression (Sox2, Nanog, Oct3/4) was comparable to BM-MSCs. NB-MSC in vitro osteogenic and chondrogenic differentiation was similar to BM-MSCs, but NB-MSCs lacked adipogenic differentiation capacity. NB-MSCs reached senescence phases at a median passage of P7 (range, P5-P13). NB-MSCs exhibited greater immunosuppressive capacity on activated T lymphocytes at a 1:2 (MSC: PBMC) ratio compared with BM-MSCs (p = 0.018). NK cytotoxic activity was not influenced by co-culture, either with BM-MSCs or NB-MSCs. Flow-cytometry cell cycle analysis showed that NB-MSCs had an increased number of cells in the G0-G1 phase compared to BM-MSCs. Transcriptomic profiling results indicated that NB-MSCs were enriched with EMT genes compared to BM-MSCs. Conclusions: We characterized the biological features, the immunomodulatory capacity and the gene expression profile of NB-MSCs. The NB-MSC gene expression profile and their functional properties suggest a potential role in promoting tumor escape, invasiveness and metastatic traits of NB cancer cells. A better understanding of the complex mechanisms underlying the interactions between NB cells and NB-derived MSCs should shed new light on potential novel therapeutic approaches