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Additional file 5: The gene specific primers were selected from the literature or were designed using Primer-BLAST software with the gene sequences obtained from the GenBank of the National Center for Biotechnology Information. NP: accession not provided.

Objectives: For decades, the detection of anti-HLA antibodies in candidates for solid-organ transplant has been performed with the traditional complement-dependent cytotoxicity method; this assay has been then integrated with the introduction of solid-phase assays. Over the past 20 years, the Luminex assay has become the most widely used in clinical laboratories due to both increased sensitivity and specificity versus enzyme-linked immunosorbent assay. However, even the Luminex technique has shown some critical issues, and choosing the most reliable method still remains challenging. In this study, we verified the concordance of the results obtained in detecting anti-HLA antibodies with 2 kit vendors that provide reagents for the Luminex platform. Materials and Methods: We used 314 serum samples from patients on wait lists for solid-organ transplant. Sera were tested with LABScreen Mixed-LSM12 (One Lambda-Thermo Fisher, Canoga Park, CA, USA) and LIFECODES LifeScreen Deluxe-LMX (Gen-Probe-Immucor, Stanford, CT, USA),which we indicated as vendor A and vendor B, respectively. Anti-HLA class I and class II antibody analyses were conducted by verifying the concordance of the results with Cohen kappa coefficient statistics and confidence interval. Results: The kappa coefficient statistics showed “substantial” reliability for class I (0.61; confidence interval, 0.50-0.73) and “moderate” reliability for class II (0.56; confidence interval, 0.43-0.69). There were no considerable differences in results between the 2 kits regarding overall assignment of negativity or positivity of a sample. Discordant data between positive values for a test and negative for the other were found for samples with weak antibody positivity. Conclusions: Some discordant data were probably attributable to several factors such as the composition of the kits, the antibody titer in the serum, whether sera were diluted, different washing methods, and type of plate used.

This doctoral dissertation is concerned with an exploratory study on how emerging learning ecologies enabled by Web 2.0 and social web are affecting the self-organized practices and dispositions in the digital settings of individual PhD students. The research endorses a constructivist grounded theory approach, where data collection has been undertaken across three Italian and one UK universities and has included a sequence of online questionnaires, individual interviews and focus groups. The findings being generated provide a repertoire of social media practices for research purposes; a framework conceptualizing the trajectories in the digital, in terms of Space, Time, Socialization, Digital identity, Stance and Tensions; the forms of resilience and the tensions underlying the PhD researchers’ digital engagement. The affordances of PhD e-researchers’ emerging ecologies are therefore understood as multi-dimensional and transitional trajectories intentionally undertaken by the individuals and generating a range of reactions toward the opportunities provided by the open Web.

La proteina prionica è un normale costituente dell'ospite, ubiquitaria nei tessuti dei vertebrati, di cui esistono due isoforme: la forma normale, PrPC, e la forma proteasi-resistente PrPSc specifica della malattia. La principale differenza tra le due è solo conformazionale, in quanto la PrPC contiene esclusivamente alfa eliche, mentre la PrPSc contiene filamenti beta. La malattia da prioni è spiegata dalla teoria prionica, la quale ipotizza che la PrPC sia in grado di ripiegarsi in modo anomalo, originando così il prione patogeno PrPSc. Quest’ultimo ha la capacità di autoreplicarsi e polimerizzare, formando placche amiloidotiche che precipitano sulla superficie dei neuroni provocandone la morte. Sulla base di pregressi studi di relazione struttura-attività, è stata sintetizzata una serie di composti, potenzialmente capaci di inibire la conversione della PrPC in PrPSc. Tali composti sono caratterizzati dalla presenza di un nucleo di varia natura (fenossibenzenico, cicloesilossibenzenico, ecc..), opportunamente sostituito con gruppi con diverse proprietà steriche ed elettroniche, legato, mediante una porzione acetammidica, ad un anello pirrolidinico o piperidinico, e recante una catena alchilica di diversa lunghezza. La sperimentazione farmacologica ha previsto un saggio HTS basato sulla scansione di target intensamente fluorescenti (SIFT, scanning for intensely fluorescent targets) al fine di valutare la capacità dei composti di interferire con l’interazione PrPC-PrPSc a livello molecolare. Successivamente è stata valutata la loro capacità di inibire la propagazione di PrPSc in sistemi cellulari (cellule ScN2a e SMB). L’attività antiprionica è stata riscontrata per la maggior parte dei composti sintetizzati; i dati sono, inoltre, caratterizzati da una significativa concordanza tra i risultati forniti dai due tipi di saggio.

Significance Chronic obstructive pulmonary disease (COPD) is an inflammatory disease characterized by limitation of expiratory airflow. Cellular and molecular pathways involved in disease pathogenesis are not completely defined. Our study reveals that metabolism and immune response cooperate in COPD pathogenesis and progression. COPD subjects with different disease stages showed progressive increase of systemic leptin, an adipose tissue-derived proinflammatory molecule, that, at high concentrations, impaired the capacity of T cells to engage in glycolysis and to generate regulatory T cells. Thus, the loss of these immunoregulatory circuits during COPD determined the hyperactivation of effector T cells that amplified inflammation, leading to progressive decline of lung function. Understanding these immunometabolic mechanisms can have important implications for monitoring COPD progression and for disease treatment., Chronic obstructive pulmonary disease (COPD) is an inflammatory condition associated with abnormal immune responses, leading to airflow obstruction. Lungs of COPD subjects show accumulation of proinflammatory T helper (Th) 1 and Th17 cells resembling that of autoreactive immune responses. As regulatory T (Treg) cells play a central role in the control of autoimmune responses and their generation and function are controlled by the adipocytokine leptin, we herein investigated the association among systemic leptin overproduction, reduced engagement of glycolysis in T cells, and reduced peripheral frequency of Treg cells in different COPD stages. These phenomena were also associated with an impaired capacity to generate inducible Treg (iTreg) cells from conventional T (Tconv) cells. At the molecular level, we found that leptin inhibited the expression of forkhead-boxP3 (FoxP3) and its splicing variants containing the exon 2 (FoxP3-E2) that correlated inversely with inflammation and weakened lung function during COPD progression. Our data reveal that the immunometabolic pathomechanism leading to COPD progression is characterized by leptin overproduction, a decline in the expression of FoxP3 splicing forms, and an impairment in Treg cell generation and function. These results have potential implications for better understanding the autoimmune-like nature of COPD and the pathogenic events leading to lung damage.

Purpose Human leukocyte antigen (HLA)-G is a non-classic major histocompatibility complex HLA class I molecule. HLA-G may have tolerogenic properties which are linked to epigenetic-sensitive pathways. There is a correlation of sHLA-G levels and graft acceptance in transplantation studies. There are previous data on correlation of sHLA-G with graft rejection as well as with viral infections such as hepatitis C virus (HCV) in kidney transplanted patients. Here, we report the sHLA-G expression in patients on the waiting list for kidney transplantation, with and without anti-HCV compared to a control group. Methods Serum of 67 patients on the waiting list for kidney transplantation (n = 43 with anti-HCV and n = 24 without anti-HCV) was analyzed. Among these patients, n = 39 were on the waiting list for the first transplantation, while n = 28 were patients who returned in the list. The control group included n = 23 blood donors with anti-HCV (n = 13) and without anti-HCV (n = 10). Results The expression of sHLA-G was significantly lower in the control group (39.6 ± 34.1 U/ml) compared to both - patients on the waiting list for the first transplantation (62.5 ± 42.4 U/ml, p=0.031) and patients who returned in the list (76.7 ± 53.9 U/ml, p=0.006). No significant differences were observed in all anti-HCV positive groups. A positive linear correlation between sHLA-G and TNF-α, and patient age was observed. Conclusions Serum sHLA-G values were significantly increased in both - patients on the waiting list for the first transplantation and patients who returned in the list, as compared to control group. Our findings confirm the key tolerogenic role of sHLA-G levels as epigenetic-related marker for measuring the state of kidney allograft acceptance.

BACKGROUND: In epileptic patients with motor disability, it's difficult to disentangle the effects of antiepileptic drugs (AEDs) on bone health from those provoked by impaired mobility. The aim of this study was to evaluate the effects of AEDs on bone mineral status by phalangeal quantitative ultrasound (QUS), a no-radiation and non-invasive method, in pediatric patients with motor impairment and epilepsy. METHODS: We enrolled 56 patients (31 females, 25 males) with epilepsy and motor impairment and 24 children with only motor disability (13 females, 11 males). Patients were stratified by Gross Motor Function Classification System Scale (GMFCS) in 4 groups: group A1 with epilepsy and mild motor impairment (GMFCS levels I-II), group A2 with only mild motor impairment, group B1 with epilepsy and severe motor impairment (GMFCS levels III-V), group B2 with only severe motor impairment. The bone mineral status was evaluated by QUS and amplitude-dependent speed of sound (AD-SoS) Z-score was calculated for each patient. RESULTS: The four groups showed no significant differences in age, gender and 25- hydroxyvitamin D levels. The group B1 had a statistically lower amplitude-dependent speed of sound Z-score as compared to group A2 (p

The medical field is becoming the sphere of a quite peculiar confrontation, a sort of test of tension between two different approaches: on one side self-referencing, the concept that the whole set of knowledge and capabilities generated by medical science establishes self-sufficiency of our field; on the other, the quest to broaden our concept of reason overcoming this self-imposed limitation of reason. At stake is the need to overcome the dissatisfaction and the crisis our field is currently going through. This article will go over some aspects of what we believe to be a paradigm of a much general and crucial cultural problem.