Μελέτη της in vitro κυτταρολυτικής δραστικότητας των ΝΚ κυττάρων ασθενών πασχόντων από Μυελικές Κακοήθειες

Στην παρούσα προοπτική μελέτη εκτιμήθηκε η συσχέτιση μεταξύ της ΝΚ κυτταροτοξικής δραστικότητας, των Μυελικών Κατασταλτικών κυττάρων (Myeloid Derived Suppressor cells - MDSCs) και των Τ-ρυθμιστικών κυττάρων (T-Regulatory cells – Tregs), ως προς τη διάρκεια της ανταπόκρισης και την ολική επιβίωση, ασθενών πασχόντων από υψηλού κινδύνου Μυελοδυσπλαστικό σύνδρομο (ΜΔΣ) ή ολιγοβλαστική Οξεία Μυελογενή Λευχαιμία (ΟΜΛ) που λάμβαναν θεραπεία με τον υπομεθυλιωτικό παράγοντα 5-Αζασυτιδίνη. Ο απώτερος σκοπός της μελέτης ήταν η αναγνώριση πρώιμων βιοδεικτών με προγνωστική σημασία ως προς την συνολική ανταπόκριση των ασθενών αυτών στη χορηγούμενη θεραπεία. Οι ασθενείς που συμμετείχαν στην παρούσα μελέτη είχαν ήδη λάβει τουλάχιστον τρείς κύκλους θεραπείας με τον υπομεθυλιωτικό παράγοντα 5-Αζασυτιδίνη. Η in vitro προσδιορισμένη ΝΚ-κυτταροτοξικότητα σε αυτή την ομάδα των ασθενών έχει προγνωστική αξία ως προς τη διάρκεια της ανταπόκρισης και ως προς την ολική επιβίωση, οι ασθενείς που επιδείκνυαν υψηλή ΝΚ-κυτταροτοξικότητα παρουσίαζαν μακρύτερη επιβίωση και μακρύτερη διάρκεια ανταπόκρισης σε σύγκριση με τους ασθενείς που επιδείκνυαν χαμηλή ΝΚ-κυτταροτοξικότητα. Οι πληθυσμοί των ανοσορρυθμιστικών κυττάρων MDSCs και Tregs στο περιφερικό αίμα ασθενών μετά βραχεία έκθεση σε 5-Αζασυτιδίνη, δε διέφεραν στατιστικώς σημαντικά σε σύγκριση με τον πληθυσμό των υγιών μαρτύρων. Σύμφωνα με τα αποτελέσματα της μελέτης μας, η θεραπευτική δράση της 5-Αζασυτιδίνης διενεργείται, τουλάχιστον μερικώς, μέσω ανοσορρυθμιστικής επίδρασης του παράγοντα αυτού. Με βάση την πρόσφατη αναζήτηση της βιβλιογραφίας, η μελέτη αυτή αποτελεί την πρώτη δημοσιευμένη μελέτη που καταγράφει θετική συσχέτιση μεταξύ της ΝΚ-κυτταροτοξικότητας και της ολικής επιβίωσης ασθενών υπό 5-Αζασυτιδίνη.In our study, we prospectively examined the correlation between NKcytotoxic activity, Myeloid-derived Suppressor cells (MDSCs) and TRegulatory cells (Tregs), with the duration of response (DOR) and the overall survival (OS) of patients with high-risk myelodysplastic syndrome (MDS) and oligoblastic acute myeloid leukaemia (AML) treated with 5-Αzacytidine (AZA), with the aim to identify early biomarkers predictive of the final outcome. Patients enrolled at the study had already received at least three cycles of 5-Αzacytidine treatment. Peripheral blood NKcytotoxicity at completion of chemotherapy is predictive of DOR and OS; AZA-treated patients with high NK-cytotoxicity survived longer and response to treatment was sustained for a longer period than patients with low NK-cytotoxicity. In contrast, numbers of MDSCs and Tregs in the peripheral blood (PB) of patients after a short exposure to 5-Αzacytidine were not significantly different from normal donors. The results of our study suggest that the therapeutic activity of 5-Αzacytidine is at least partly mediated by an immunomodulatory effect. To our knowledge, this is the first study that shows a positive correlation between NK-cytotoxicity and OS of AZA-treated patients

Anti-Ro60 Seropositivity Determines Anti-Ro52 Epitope Mapping in Patients With Systemic Sclerosis

Epitope mapping of anti-Ro52 antibodies (Abs) has been extensively studied in patients with Sjögren's syndrome (SjS) and systemic lupus erythematosus (SLE). Comprehensive epitope mapping in systemic sclerosis (SSc), where anti-Ro52 antibodies are also frequently detected, has not been performed. The aim of the present study was to fully characterize Ro52 epitopes in anti-Ro52-positive SSc using Ro52 fragments spanning the full antigen. Further analysis was made according to anti-Ro60 status. Epitope mapping was performed in 43 anti-Ro52-positive SSc patients. Seventy eight anti-Ro52-positive pathological controls, including 20 patients with SjS, 28 patients with SLE, 15 patients with dermatomyositis (DM), and 15 patients with primary biliary cholangitis (PBC), and 20 anti-Ro52-negative healthy individuals as normal controls were also tested. Five recombinant Ro52 fragments [Ro52-1 (aa 1-127), Ro52-2 (aa 125-268), Ro52-3 (aa 268-475), Ro52-4 (aa 57-180), and Ro52-5 (aa 181-320) were used to test reactivity by line-immunoassay and in house ELISA. Anti-Ro60 reactivity was tested by ELISA. All anti-Ro52 positive sera reacted with Ro52-2; none recognized Ro52-3. Antibodies against Ro52-1 were less frequently found in SSc than in SjS/SLE (11.6 vs. 41.7%, p = 0.001); and antibodies against Ro52-4 were less frequently found in SSc than in SjS/SLE (27.9 vs. 50%, p = 0.03). In SSc patients, reactivity against Ro52-1 was more frequent in anti-Ro52+/anti-Ro60+ than in anti-Ro52+/anti-Ro60-patients (33.3 vs. 0%, p = 0.003). In this comprehensive analysis of Ro52 epitope mapping in SSc, the coiled coil domain remains the predominant epitope on Ro52. Contrary to SjS and SLE, patients with SSc fail to identify epitopic regions within the N-terminus of the protein, especially if they lack con-current anti-Ro60 reactivity

Could Immunophenotype Guide Molecular Analysis in Patients with Myeloid Malignancies?

Objective: Immunophenotype has been correlated with molecular aberrations in several studies. The aim of this study was the discovery of immunophenotypic features related to mutations in AML and MDS patients connected to prognostic factors. Moreover, an effort to evaluate a method for the detection of the most common NPM1 mutations of exon12 and Internal Tandem Duplications (ITD) mutations of FLT3 gene by flow cytometry was performed. Method: Patients with de novo myeloid neoplasms [ AML and MDS (AML-M3 patients were excluded)] were included. FLT3/ITD/TKD and NPM1 mutations were detected by PCR and fragment analysis. The immunophenotypic analysis was performed by multi-dimensional flow cytometry (FC) with a standardized panel of monoclonal antibodies on peripheral blood or bone marrow samples. Nucleophosmin Antibody and CD135 were used for the mutations immunophenotypic detection. Results: NPM1 and/or FLT3 mutations correlated with low or no expression of more immature cells markers such as CD34, CD117, HLADR, as well as higher expression of more mature markers such as CD11b. The higher expression of CD33 should be mentioned as well. The presence of NPM1mut and FLT3/ITD does not seem to be detectable by FC at least using these two monoclonal antibodies. The presence of CD7 aberrant lymphoid marker’s expression was associated with FLT3mut, NPM1wt genotype. CD56 or CD2 positivity was found only in patients’ samples negative for NPM1 and/or FLT3 mutations. Conclusions: Certain immunophenotype findings including the presence of aberrant lymphoid markers may be indicative of the presence of mutations in NPM1 and FLT3 linked to prognosis

Global patient outcomes after elective surgery: prospective cohort study in 27 low-, middle- and high-income countries.

BACKGROUND: As global initiatives increase patient access to surgical treatments, there remains a need to understand the adverse effects of surgery and define appropriate levels of perioperative care. METHODS: We designed a prospective international 7-day cohort study of outcomes following elective adult inpatient surgery in 27 countries. The primary outcome was in-hospital complications. Secondary outcomes were death following a complication (failure to rescue) and death in hospital. Process measures were admission to critical care immediately after surgery or to treat a complication and duration of hospital stay. A single definition of critical care was used for all countries. RESULTS: A total of 474 hospitals in 19 high-, 7 middle- and 1 low-income country were included in the primary analysis. Data included 44 814 patients with a median hospital stay of 4 (range 2-7) days. A total of 7508 patients (16.8%) developed one or more postoperative complication and 207 died (0.5%). The overall mortality among patients who developed complications was 2.8%. Mortality following complications ranged from 2.4% for pulmonary embolism to 43.9% for cardiac arrest. A total of 4360 (9.7%) patients were admitted to a critical care unit as routine immediately after surgery, of whom 2198 (50.4%) developed a complication, with 105 (2.4%) deaths. A total of 1233 patients (16.4%) were admitted to a critical care unit to treat complications, with 119 (9.7%) deaths. Despite lower baseline risk, outcomes were similar in low- and middle-income compared with high-income countries. CONCLUSIONS: Poor patient outcomes are common after inpatient surgery. Global initiatives to increase access to surgical treatments should also address the need for safe perioperative care. STUDY REGISTRATION: ISRCTN5181700

The state of play in European coaching & mentoring

This report provides an overview of the main findings from the 2017 European Coaching and Mentoring Research Project, undertaken by Jonathan Passmore and Hazel Brown, in partnership with the EMCC and the wider European coaching and mentoring industry. The study was planned in 2016 and undertaken during a 12-week period, between March and May 2017. This is one of a number reports published. This Executive Report is available free of charge, along with a National Report in countries that achieved over 50 coach or mentor participants. Each National Report is published in the language chosen by of the respective national coaching community. The aim of these national reports is to deepen understanding of coaching and mentoring and to widen engagement with coaching and mentoring.peer-reviewe

In vitro NK - cytolytic activity of patients suffering from myeloid malignancies

In our study, we prospectively examined the correlation between NK- cytotoxic activity, Myeloid-derived Suppressor cells (MDSCs) and T-Regulatory cells (Tregs), with the duration of response (DOR) and the overall survival (OS) of patients with high-risk myelodysplastic syndrome (MDS) and oligoblastic acute myeloid leukaemia (AML) treated with 5-Αzacytidine (AZA), with the aim to identify early biomarkers predictive of the final outcome. Patients enrolled at the study had already received at least three cycles of 5-Αzacytidine treatment. Peripheral blood NK-cytotoxicity at completion of chemotherapy is predictive of DOR and OS; AZA-treated patients with high NK-cytotoxicity survived longer and response to treatment was sustained for a longer period than patients with low NK-cytotoxicity. In contrast, numbers of MDSCs and Tregs in the peripheral blood (PB) of patients after a short exposure to 5-Αzacytidine were not significantly different from normal donors. The results of our study suggest that the therapeutic activity of 5-Αzacytidine is at least partly mediated by an immunomodulatory effect. To our knowledge, this is the first study that shows a positive correlation between NK-cytotoxicity and OS of AZA-treated patients.Στην παρούσα προοπτική μελέτη εκτιμήθηκε η συσχέτιση μεταξύ της ΝΚ κυτταροτοξικής δραστικότητας, των Μυελικών Κατασταλτικών κυττάρων (Myeloid Derived Suppressor cells - MDSCs) και των Τ-ρυθμιστικών κυττάρων (T-Regulatory cells – Tregs), ως προς τη διάρκεια της ανταπόκρισης και την ολική επιβίωση, ασθενών πασχόντων απο υψηλού κινδύνου Μυελοδυσπλαστικό σύνδρομο (ΜΔΣ) ή ολιγοβλαστική Οξεία Μυελογενή Λευχαιμία (ΟΜΛ) που λάμβαναν θεραπεία με τον υπομεθυλιωτικό παράγοντα 5-Αζασυτιδίνη. Ο απώτερος σκοπός της μελέτης ήταν η αναγνώριση πρώϊμων βιοδεικτών με προγνωστική σημασία ως προς την συνολική ανταπόκριση των ασθενών αυτών στη χορηγούμενη θεραπεία.Οι ασθενείς που συμμετείχαν στην παρούσα μελέτη είχαν ήδη λάβει τουλάχιστον τρείς κύκλους θεραπείας με τον υπομεθυλιωτικό παράγοντα 5-Αζασυτιδίνη. Η in vitro προσδιορισμένη ΝΚ-κυτταροτοξικότητα σε αυτή την ομάδα των ασθενών έχει προγνωστική αξία ως προς τη διάρκεια της ανταπόκρισης και ως προς την ολική επιβίωση, οι ασθενείς που επιδείκνυαν υψηλή ΝΚ-κυτταροτοξικότητα παρουσίαζαν μακρύτερη επιβίωση και μακρύτερη διάρκεια ανταπόκρισης σε σύγκριση με τους ασθενείς που επιδείκνυαν χαμηλή ΝΚ-κυτταροτοξικότητα. Οι πληθυσμοί των ανοσορρυθμιστικών κυττάρων MDSCs και Tregs στο περιφερικό αίμα ασθενών μετα βραχεία έκθεση σε 5-Αζασυτιδίνη, δε διέφεραν στατιστικώς σημαντικά σε σύγκριση με τον πληθυσμό των υγιών μαρτύρων.Σύμφωνα με τα αποτελέσματα της μελέτης μας, η θεραπευτική δράση της 5-Αζασυτιδίνης διενεργείται, τουλάχιστον μερικώς, μέσω ανοσορρυθμιστικής επίδρασης του παράγοντα αυτού. Με βάση την πρόσφατη αναζήτηση της βιβλιογραφίας, η μελέτη αυτή αποτελεί την πρώτη δημοσιευμένη μελέτη που καταγράφει θετική συσχέτιση μεταξύ της ΝΚ-κυτταροτοξικότητας και της ολικής επιβίωσης ασθενών υπο 5-Αζασυτιδίνη

Wound heating: Immunological aspects

Wound heating is a complex biological process, comprised of a series of a sequential events aiming to repair injured tissue. The role of the immune system in this process is not only to recognise and combat the newly presented antigens at the site of injury, but also to participate in the debridement of the damaged area and to contribute to the process of heating. In this review, we discuss the molecules and cells of the immune system that participate in tissue repair. We describe the mechanisms of immune recognition during initial insult and the innate and adaptive immune responses to injury. Finally, we address the role of the immune system in regeneration and repair. (C) 2006 Elsevier Ltd. All rights reserved

Clinical Value of Plasma Soluble Urokinase-Type Plasminogen Activator Receptor Levels in Term Neonates with Infection or Sepsis: A Prospective Study

Background. suPAR, the soluble form of the urokinase-type plasminogen activator receptor, has been identified as a biomarker of infection in adults but its properties in neonatal infection are not known. Methods. Plasma suPAR levels were determined by ELISA in 47 term neonates with infection (19 bacterial and 28 viral) and in 18 healthy neonates as controls. Thirteen out of 47 infected neonates were septic. In all infected neonates, suPAR levels were repeated at 24 hours, 48 hours, 3-5 days, and 7-10 days following admission. Results. Plasma suPAR levels were significantly increased in infected neonates upon admission, whereas they were highest in septic neonates, in comparison with controls (P < 0.001) and correlated positively with serumCRP levels (P = 0.001). At infection subsidence, suPAR concentrations decreased significantly in comparison with baseline (P < 0.001) but remained higher than in controls (P = 0.01). Receiver operating characteristic analysis resulted in significant areas under the curve for detecting either infected or septic neonates, but not for discriminating between bacterial and viral cause of infection. Conclusions. suPAR is a diagnostic biomarker of infection or sepsis in term neonates; however, it cannot discriminate bacterial from viral infections and also its utility for monitoring the response to treatment is questioned

Clinical Value of Plasma Soluble Urokinase-Type Plasminogen Activator Receptor Levels in Term Neonates with Infection or Sepsis: A Prospective Study

Background. suPAR, the soluble form of the urokinase-type plasminogen activator receptor, has been identified as a biomarker of infection in adults but its properties in neonatal infection are not known. Methods. Plasma suPAR levels were determined by ELISA in 47 term neonates with infection (19 bacterial and 28 viral) and in 18 healthy neonates as controls. Thirteen out of 47 infected neonates were septic. In all infected neonates, suPAR levels were repeated at 24 hours, 48 hours, 3–5 days, and 7–10 days following admission. Results. Plasma suPAR levels were significantly increased in infected neonates upon admission, whereas they were highest in septic neonates, in comparison with controls P<0.001 and correlated positively with serum CRP levels (P=0.001). At infection subsidence, suPAR concentrations decreased significantly in comparison with baseline (P<0.001) but remained higher than in controls (P=0.01). Receiver operating characteristic analysis resulted in significant areas under the curve for detecting either infected or septic neonates, but not for discriminating between bacterial and viral cause of infection. Conclusions. suPAR is a diagnostic biomarker of infection or sepsis in term neonates; however, it cannot discriminate bacterial from viral infections and also its utility for monitoring the response to treatment is questioned