Background: Background: We previously demonstrated that 76% of allogeneic (ALLO) and 94% of autologous (AUTO) hematopoietic stem cell transplantation (HSCT) recipients develop a serological response after two doses of BNT162b2 anti SARS-Cov-2 mRNA vaccine (Attolico et al, BJH 2021). However, less is known about the lymphocyte (Ly) immune our preliminary evaluations show differences in Ly subsets amounts and T and B maturation profiles between HC and transplant patients. Furthermore, no changes were observed before and after the booster dose even in NR who subsequently developed an antibody response. Finally, our study confirms the predictive role of CD4+/CD8+ ratio for the response to vaccination.

Inflammatory bowel diseases (IBDs) are a group of chronic conditions of the gastrointestinal tract in which nationwide studies have revealed a higher risk of hematological malignancies (HMs). Clonal hematopoiesis (CH) is a premalignant condition defined by the presence of an acquired somatic mutation characterized by a variant allele frequency (VAF) of ≥2%, in a gene frequently associated with HMs. A growing body of evidence suggests a correlation between inflammation and CH; its occurrence in the context of IBD has been previously demonstrated. With the aim to assess CH possible co-occurrence in patients with an IBD associated with HMs, we performed a targeted next-generation sequencing analysis in a cohort of thirteen patients who were referred to our center with IBD associated with HMs. Eleven (85%) patients showed one or more mutations in CH-associated genes; DNMT3A was the most frequently mutated gene, followed by ASXL1 and JAK2. These results may suggest that the mechanisms at the basis of the inflammatory environment could potentially select for the growth of hematopoietic clones harboring specific mutations. In this context, CH emergence may be boosted by the proinflammatory IBD environment, thus acting as a biological link between IBD and the HM onset. If these data are confirmed, IBD patients screened and positive for CH should undergo a hematologic follow-up to assess the risk of developing HM. Future study will clarify the relationship between these conditions.

Introduction. The JAK2 haplotype known as "GGCC or 46/1 haplotype" consists of a germline combination of single nucleotide polymorphisms (SNPs) that are inherited together and are frequently associated with the onset of myeloproliferative neoplasms (MPNs) positive for both JAK2 V617 and exon 12 mutations. It has been reported a significant association between JAK2 negative erythrocytosis and the simultaneous occurrence of JAK2 haplotype GGCC_46/1 and CALR rs1049481_G allele, mapping in the 3' UTR of the gene. In the present study, we investigated the presence of JAK2 haplotype GGCC_46/1 and CALR rs1049481_G allele in a more extensive series of erythrocytosis patients and evaluated a possible correlation with serum erythropoietin (EPO) level. Methods. Fifty-nine erythrocytosis patients, diagnosed at our Center from 2009 to 2020, negative for canonical JAK2 mutations, and secondary causes, were included in this study. Forty-nine (83%) out of 59 cases met only two major WHO 2016 diagnostic criteria for polycythemia vera (PV); the remaining 10 cases were "JAK2 negative PV", as they also showed the minor criterion with subnormal serum EPO. Genomic DNA from all patients was extracted from peripheral blood granulocytes and analyzed for the presence of both SNPs. The occurrence of the JAK2 haplotype was investigated by analyzing rs10974944 tagging SNP, using distal JAK2 intron 12 primers (forward: 5'-CTGTGCAGTCCAAACCCATG-3' and reverse: 5'-TTCTCTGCTTGCTAGTGGGT-3'). This amplification generated a PCR product of 272 bp, revealing a C/G substitution by Sanger sequencing. PCR investigated the occurrence of the G/T allele in the SNP rs1049481 in the CALR gene with primers CALR-UTR-F 5'-GACAAGGAGGATGATGAGGACAAA-3' and CALR-UTR-R 5'- AAAAATGAAAGTTCTCGAGTCTCACAGA-3' generating a 205 bp product. In silico data from 2504 healthy individuals of the 1000G Project (1000G) were used as a control group. Fisher's exact test was used to compare the differences of both SNPs and the frequency of their alleles between cases and controls, respectively; the c-square test was employed for evaluating the distribution of JAK2 GGCC haplotype in erythrocytosis patients and controls. Results. Thirty-eight (64.4%) and 21 (35.6%) cases resulted in being positive and negative for the JAK2 GGCC haplotype, respectively. The JAK2 GGCC haplotype occurred to be associated with erythrocytosis as a statistically significant difference in frequency was detected as respect to 2504 healthy individuals from the 1000G Project (p=0.0013). This finding was also confirmed considering the Genome Aggregation Database (gnomAD v2.1.1, https://gnomad.broadinstitute.org/)(p=0.0024). The association was also demonstrated in terms of allelic frequency (p=0.0131) and genotype distribution (p=0.0030). Regarding CALR rs1049481 SNP, a significant difference in the CALR rs1049481_G allelic rate was confirmed in our cohort compared to 1000G controls (p=0.0360). Overall, the association between JAK2 GGCC haplotype and CALR rs1049481_G was observed in 28/59 (47.5%) erythrocytosis cases compared to healthy individuals (628/2504, 25%), and this difference resulted in being statistically significant (p=0.0002). Based on the EPO level, erythrocytosis patients were divided into two groups: normal (49 cases) or subnormal (10 cases). Interestingly, the simultaneous presence of JAK2 GGCC haplotype and CALR rs1049481_G was statistically significantly associated with the erythrocytosis group showing normal EPO (p=0.0133). Conclusions. This study suggests that the JAK2 GGCC haplotype and the presence of the CALR rs1049481_G allele were significantly associated with erythrocytosis cases, negative for canonical JAK2 mutations. Our findings are in line with recent literature evidence showing that germline predisposition factors such as SNPs and haplotypes may play a crucial role in MPN pathogenesis. Moreover, this study shows that patients showing two major WHO 2016 diagnostic criteria (erythrocytosis and panmyelosis) without JAK2 mutations and with normal EPO levels can benefit from the search for germline polymorphisms combination in JAK2 and CALR driver genes for a better diagnostic classification. Therefore, the presence of these polymorphisms could represent a novel minor criterion for the diagnosis of "JAK2 negative PV". Disclosures No relevant conflicts of interest to declare.

Background:Acute Myeloid Leukemia is the most common acute leukemia in adults; median age at diagnosis is around 68 years. AML is cured in 35 to 40% of adult patients <65 yrs and in 5 to 15% of pts >70 yrs. The number of AML survivors is growing and is relevant the evaluation of health related quality of life associated with disease and treatment to propose potential strategies for intervention.Aims:We collected data on 382 AML adult pts (age<65 yrs) treated in our center. Of these, 77(20.2%) are long term survivors (LTS) (OS>5yrs). We evaluated the QoL of this subset of patients using the EORTC‐QLQ‐C30 questionnaire.Methods:From 2003 to 2013, 382 adult pts (age<65 yrs) were diagnosed with AML in our center. According to Revised Reccomendations (Cheson 2003) and ELN 2010 risk categories, 89 (23.3%) pts were at high risk, 215 (56.3%) intermediate and 78 (20.4%) low risk; 77 (20.2%)pts are long term survivors (OS>5 yrs). In this pts subset median age at diagnosis was 41.5 yrs (range 18–62). According to ELN 2010 risk categories only 7 (9%) were at HR, 36 (46.9%) IR and 34 (44.1%) LR; 42 (54.5%) pts underwent Allo‐HSCT, 13 (16.9%) Auto‐HSCT and 22 (28.6%) were treated with conventional consolidation regimens. OS was >5 yrs in 43 (55.8%) pts, and >10 yrs in 34 (44.2%).Our study included 35/77 (45,4%) randomly selected long term survivors;21(60%) underwent Allo‐HSCT, 8 (22.8%) Auto‐HSCT and 6 (17.2%) were treated with conventional CHT. Relapse occurred in 10 (28.6%) pts. Only 4 (11.4%) had “primary refractory” disease. 21/35 (60%) have an OS>5 yrs and 14/35 (40%) patients an OS>10 yrs. We compared data obtained from our sample to European reference values for the EORTC QLQ‐C30 questionnaire in order to establish factors affecting the QoL of LTS AML pts.Results:Global QoL of AML LTS pts is equal to that in the general population, but a reduction of all values in both functioning and symptoms scales is present. In the functioning scales minimum clinically significant differences (MCID:10 points) are present in the role, cognitive and social indexes. In the symptoms scales, insomnia, constipation and financial difficulties indexes reached the MCID. In order to establish which factors have a stronger impact on the QoL of AML pts, we stratified our sample considering these features: age at diagnosis (cut‐off: 55 years), Allo‐HSCT, OS>10 yrs. Age at diagnosis has an impact on symptoms scales: older patients have a clinically significant difference in fatigue, dyspnea and pain. QoL of younger pts is better but does not reach MCID. Pts who underwent Allo‐HSCT have a worse global QoL and clinically significant differences in symptoms scales, particularly dyspnea, pain, insomnia and fatigue. Pts with an OS>10 yrs have a global QoL and both functional and symptoms scales values equal to that of the general population (IMG 1). Differences in symptoms such as pain, fatigue, dyspnea, are present in patients with an OS<10 yrs.Summary/Conclusion:LTS AML pts are still a minority; with advances in therapeutic procedures they will hopefully grow. Therefore, QoL evaluation is a critical aspect to watch for. Our study suggests that age, disease characteristics and Allo‐HSCT have a strong impact on AML pts QoL. However, pts with an OS>10 years may be considered completely “cured” as their QoL values are equal to those in the general population. A final consideration should be made regarding the reintegration of AML pts in appropriate social and economic contexts, as financial difficulties were one of the most clinically significant values in all questionnaires.image

The relationship between left ventricular ejection fraction (LVEF) and cognitive performance in patients with coronary artery disease (CAD) without overt heart failure is still under debate. In the present study we combine behavioural measures and event-related potentials (ERPs) to verify whether electrophysiological correlates of recognition memory (Old/New effect) is modulated differently as a function of LVEF. Twenty-five male patients with CAD (13 without [LVEF > 55%] and 12 with [LVEF < 40%] left ventricular dysfunction), and a Mini Mental State Examination score > 25 were enrolled. ERPs were recorded while participants performed a visual word recognition task consisting of a test phase in which they were asked to judge whether visual stimuli were previously presented in a learning phase (‘old’) or not (‘new’). ERPs responses from the test phase were analyzed. A late positive ERP component between 300 and 500 ms was differentially modulated in the two groups: a clear old/new effect (enhanced mean amplitude for old respect to new items) was observed in patients without LVEF dysfunction; whereas patients with overt LVEF dysfunction did not show such old/new effect. These data suggest that ERPs may reveal possible functional brain abnormalities that might be not observed at behavioural levels.

BACKGROUND:Epicardial adipose tissue (EAT) is a visceral fat that fulfills two important functions: lipid-storage and secretion of adipokines with pro-inflammatory and pro-atherogenic properties. It has been suggested that EAT may affect the pathogenesis of atherosclerosis and the clinical course of coronary artery disease (CAD). In patients with obesity, diabetes and metabolic syndrome, the epicardial adipose tissue is enlarged. Little is known about the role of EAT in left ventricular dysfunction. Aim of this study was to evaluate the ability of insulin resistance to predict EAT thickness in patients with significant CAD and systolic dysfunction.METHODS:We enrolled 114 subjects diagnosed with CAD by angiography. The majority underwent revascularization after an acute coronary syndrome. Patients were considered affected by significant left ventricular dysfunction when EF was < or = 40%. Three indexes of insulin resistance, the HOMA IR index, the insulin sensitivity QUICKI index, and the novel adiponectin/resistin index (ADIPO-IRAR) were calculated and correlated to EAT thickness. Epicardial fat was measured by echocardiography according to standardized methods.RESULTS:Subjects with diabetes and with a history of hypercholesterolemia had thicker EAT compared to controls. Potassium levels and all three indexes of insulin resistance were the best independent predictors of EAT in the study population as a whole and in the subset of patients with left ventricular dysfunction. In the latter group the novel ADIPO-IRAR index displayed the strongest predictivity.CONCLUSION:Insulin resistance is an independent predictor of EAT thickness in patients affected by CAD, also in the presence of significant left ventricular dysfunction.

BACKGROUND AND AIMS:Chronic heart failure (HF) is characterised by a neurohormonal dysfunction associated with chronic inflammation. A role of metabolic derangement in the pathophysiology of HF has been recently reported. Adiponectin, an adipose-tissue-derived cytokine, seems to play an important role in cardiac dysfunction. We investigated the variation of circulating adiponectin in patients with coronary artery disease (CAD), with or without HF, in order to identify its independent predictors. METHODS AND RESULTS:A total of 107 outpatients with CAD were enrolled in the study and divided into three groups: CAD without left ventricular systolic dysfunction (group 1); CAD with left ventricular dysfunction without HF symptoms (group 2) and CAD with overt HF (group 3). Plasma adiponectin was determined by enzyme-linked immunosorbent assay. Adiponectin concentrations increased progressively from group 1 (7.6 ± 3.6 ng ml⁻¹) to group 2 (9.1 ± 6.7 ng ml⁻¹) and group 3 (13.7 ± 7.6 ng ml⁻¹), with the difference reaching statistical significance in group 3 versus 1 and 2 (p < 0.001). A multivariable model of analysis demonstrated that the best predictors of plasma adiponectin were body mass index, N-terminal pro-brain natriuretic peptide and high-density lipoprotein cholesterol. However, even after adjusting for all three independent predictors, the increase of adiponectin in group 3 still remained statistically significant (p = 0.015). CONCLUSION:Our data confirm the rise of adiponectin in overt HF. The levels of circulating adipokine seem to be mainly predicted by the metabolic profile of patients and by biohumoral indicators, rather than by clinical and echocardiographic indexes of HF severity.

Metabolic and neurodegenerative disorders have a growing prevalence in Western countries. Available epidemiologic and neurobiological evidences support the existence of a pathophysiological link between these conditions. Glucagon-like peptide 1 (GLP-1), whose activity is reduced in insulin resistance, has been implicated in central nervous system function, including cognition, synaptic plasticity, and neurogenesis. We review the experimental researches suggesting that GLP-1 dysfunction might be a mediating factor between Type 2 diabetes mellitus (T2DM) and neurodegeneration. Drug treatments enhancing GLP-1 activity hold out hope for treatment and prevention of Alzheimer's disease (AD) and cognitive decline.

This letter is in response to the report by Zania et al. ([2006][1]). The authors used in vitro (Matrigel) and in vivo (chick chorioallantoic membrane) models of angiogenesis to demonstrate that small molecule PAR1 antagonists, SCH79797 and RWJ56110, block formation of new blood vessels through