C6orf10 low-frequency and rare variants in italian multiple sclerosis patients

In light of the complex nature of multiple sclerosis (MS) and the recently estimated contribution of low-frequency variants into disease, decoding its genetic risk components requires novel variant prioritization strategies. We selected, by reviewing MS Genome Wide Association Studies (GWAS), 107 candidate loci marked by intragenic single nucleotide polymorphisms (SNPs) with a remarkable association (p-value <= 5 x 10(-6)). A whole exome sequencing (WES)-based pilot study of SNPs with minor allele frequency (MAF) <= 0.04, conducted in three Italian families, revealed 15 exonic low-frequency SNPs with affected parent-child transmission. These variants were detected in 65/120 Italian unrelated MS patients, also in combination (22 patients). Compared with databases (controls gnomAD, dbSNP150, ExAC, Tuscany-1000 Genome), the allelic frequencies of C6orf10 rs 16870005 and IL2RA rs12722600 were significantly higher (i.e., controls gnomAD, p = 9.89 x 10(-7) and p < 1 x 10(-20)). TET2 rs61744960 and TRAF3 rs138943371 frequencies were also significantly higher, except in Tuscany-1000 Genome. Interestingly, the association of C6orf10 rs16870005 (Ala431Thr) with MS did not depend on its linkage disequilibrium with the HLA-DRB1 locus. Sequencing in the MS cohort of the C6orf10 3' region revealed 14 rare mutations (10 not previously reported). Four variants were null, and significantly more frequent than in the databases. Further, the C6orf10 rare variants were observed in combinations, both intra-locus and with other low-frequency SNPs. The C6orf10 Ser389Xfr was found homozygous in a patient with early onset of the MS. Taking into account the potentially functional impact of the identified exonic variants, their expression in combination at the protein level could provide functional insights in the heterogeneous pathogenetic mechanisms contributing to MS.In light of the complex nature of multiple sclerosis (MS) and the recently estimated contribution of low-frequency variants into disease, decoding its genetic risk components requires novel variant prioritization strategies. We selected, by reviewing MS Genome Wide Association Studies (GWAS), 107 candidate loci marked by intragenic single nucleotide polymorphisms (SNPs) with a remarkable association (p-value ≤ 5 × 10−6). A whole exome sequencing (WES)-based pilot study of SNPs with minor allele frequency (MAF) ≤ 0.04, conducted in three Italian families, revealed 15 exonic low-frequency SNPs with affected parent-child transmission. These variants were detected in 65/120 Italian unrelated MS patients, also in combination (22 patients). Compared with databases (controls gnomAD, dbSNP150, ExAC, Tuscany-1000 Genome), the allelic frequencies of C6orf10 rs16870005 and IL2RA rs12722600 were significantly higher (i.e., controls gnomAD, p = 9.89 × 10−7 and p < 1 × 10−20). TET2 rs61744960 and TRAF3 rs138943371 frequencies were also significantly higher, except in Tuscany-1000 Genome. Interestingly, the association of C6orf10 rs16870005 (Ala431Thr) with MS did not depend on its linkage disequilibrium with the HLA-DRB1 locus. Sequencing in the MS cohort of the C6orf10 3′ region revealed 14 rare mutations (10 not previously reported). Four variants were null, and significantly more frequent than in the databases. Further, the C6orf10 rare variants were observed in combinations, both intra-locus and with other low-frequency SNPs. The C6orf10 Ser389Xfr was found homozygous in a patient with early onset of the MS. Taking into account the potentially functional impact of the identified exonic variants, their expression in combination at the protein level could provide functional insights in the heterogeneous pathogenetic mechanisms contributing to MS

Severe Reduction in Number and Function of Peripheral T Cells Does Not Afford Protection toward Emphysema and Bronchial Remodeling Induced in Mice by Cigarette Smoke

8openThe protein Lck (p56(Lck)) is a Src family tyrosine kinase expressed at all stages of thymocyte development and is required for maturation of T cells. The targeted disruption of Lck gene in mice results in severe block in thymocyte maturation with substantial reduction in the development of CD4(+)CD8(+) thymocytes, severe reduction of peripheral T cells, and disruption of T-cell receptor signaling with defective function of T-cell responses. To investigate the role of T lymphocyte in the development of cigarette smoke-induced pulmonary changes, Lck(-/-) mice and corresponding congenic wild-type mice were chronically exposed to cigarette smoke, and their lungs were analyzed by biochemical, immunologic, and morphometric methods. Smoking mice from both genotypes showed disseminated foci of emphysema and large areas of goblet cell metaplasia in bronchial and bronchiolar epithelium. Morphometric evaluation of lung changes and lung elastin determination confirmed that mice from both genotypes showed the same degree of emphysematous lesions. Thus, cigarette smoke exposure in the presence of severe reduction in number and function of peripheral T cells does not influence the development of pulmonary changes induced by cigarette smoke. The data obtained suggest that innate immunity is a leading actor in the early development of pulmonary changes in smoking mice and that the adaptive immune response may play a role at later stages.openDe Cunto, Giovanna; Lunghi, Benedetta; Bartalesi, Barbara; Cavarra, Eleonora; Fineschi, Silvia; Ulivieri, Cristina; Lungarella, Giuseppe; Lucattelli, MonicaDE CUNTO, Giovanna; Lunghi, Benedetta; Bartalesi, Barbara; Cavarra, Eleonora; Fineschi, Silvia; Ulivieri, Cristina; Lungarella, Giuseppe; Lucattelli, Monic

Managing chronic myeloid leukemia for treatment-free remission: a proposal from the GIMEMA CML WP

Several papers authored by international experts have proposed recommendations on the management of BCR-ABL1+ chronic myeloid leukemia (CML). Following these recommendations, survival of CML patients has become very close to normal. The next, ambitious, step is to bring as many patients as possible into a condition of treatment-free remission (TFR). The Gruppo Italiano Malattie EMatologiche dell'Adulto (GIMEMA; Italian Group for Hematologic Diseases of the Adult) CML Working Party (WP) has developed a project aimed at selecting the treatment policies that may increase the probability of TFR, taking into account 4 variables: the need for TFR, the tyrosine kinase inhibitors (TKIs), the characteristics of leukemia, and the patient. A Delphi-like method was used to reach a consensus among the representatives of 50 centers of the CML WP. A consensus was reached on the assessment of disease risk (EUTOS Long Term Survival [ELTS] score), on the definition of the most appropriate age boundaries for the choice of first-line treatment, on the choice of the TKI for first-line treatment, and on the definition of the responses that do not require a change of the TKI (BCR-ABL1 6410% at 3 months, 641% at 6 months, 640.1% at 12 months, 640.01% at 24 months), and of the responses that require a change of the TKI, when the goal is TFR (BCR-ABL1 &gt;10% at 3 and 6 months, &gt;1% at 12 months, and &gt;0.1% at 24 months). These suggestions may help optimize the treatment strategy for TFR

Effect of roflumilast on inflammatory cells in the lungs of cigarette smoke-exposed mice

<p>Abstract</p> <p>Background</p> <p>We reported that roflumilast, a phosphodiesterase 4 inhibitor, given orally at 5 mg/kg to mice prevented the development of emphysema in a chronic model of cigarette smoke exposure, while at 1 mg/kg was ineffective. Here we investigated the effects of roflumilast on the volume density (V_V) of the inflammatory cells present in the lungs after chronic cigarette smoke exposure.</p> <p>Methods</p> <p>Slides were obtained from blocks of the previous study and V_Vwas assessed immunohistochemically and by point counting using a grid with 48 points, a 20× objective and a computer screen for a final magnification of 580×. Neutrophils were marked with myeloperoxidase antibody, macrophages with Mac-3, dendritic cells with fascin, B-lymphocytes with B220, CD4+ T-cells with CD4+ antibody, and CD8+T-cells with CD8-α. The significance of the differences was calculated using one-way analysis of variance.</p> <p>Results</p> <p>Chronic smoke exposure increased neutrophil V_Vby 97%, macrophage by 107%, dendritic cell by 217%, B-lymphocyte by 436%, CD4+ by 524%, and CD8+ by 417%. The higher dose of roflumilast prevented the increase in neutrophil V_Vby 78%, macrophage by 82%, dendritic cell by 48%, B-lymphocyte by 100%, CD4+ by 98% and CD8+ V_Vby 88%. The lower dose of roflumilast did not prevent the increase in neutrophil, macrophage and B-cell V_Vbut prevented dendritic cells by 42%, CD4+ by 55%, and CD8+ by 91%.</p> <p>Conclusion</p> <p>These results indicate (<it>i</it>) chronic exposure to cigarette smoke in mice results in a significant recruitment into the lung of inflammatory cells of both the innate and adaptive immune system; (<it>ii</it>) roflumilast at the higher dose exerts a protective effect against the recruitment of all these cells and at the lower dose against the recruitment of dendritic cells and T-lymphocytes; (<it>iii</it>) these findings underline the role of innate immunity in the development of pulmonary emphysema and (<it>iiii</it>) support previous results indicating that the inflammatory cells of the adaptive immune system do not play a central role in the development of cigarette smoke induced emphysema in mice.</p

Infected pancreatic necrosis: outcomes and clinical predictors of mortality. A post hoc analysis of the MANCTRA-1 international study

: The identification of high-risk patients in the early stages of infected pancreatic necrosis (IPN) is critical, because it could help the clinicians to adopt more effective management strategies. We conducted a post hoc analysis of the MANCTRA-1 international study to assess the association between clinical risk factors and mortality among adult patients with IPN. Univariable and multivariable logistic regression models were used to identify prognostic factors of mortality. We identified 247 consecutive patients with IPN hospitalised between January 2019 and December 2020. History of uncontrolled arterial hypertension (p = 0.032; 95% CI 1.135-15.882; aOR 4.245), qSOFA (p = 0.005; 95% CI 1.359-5.879; aOR 2.828), renal failure (p = 0.022; 95% CI 1.138-5.442; aOR 2.489), and haemodynamic failure (p = 0.018; 95% CI 1.184-5.978; aOR 2.661), were identified as independent predictors of mortality in IPN patients. Cholangitis (p = 0.003; 95% CI 1.598-9.930; aOR 3.983), abdominal compartment syndrome (p = 0.032; 95% CI 1.090-6.967; aOR 2.735), and gastrointestinal/intra-abdominal bleeding (p = 0.009; 95% CI 1.286-5.712; aOR 2.710) were independently associated with the risk of mortality. Upfront open surgical necrosectomy was strongly associated with the risk of mortality (p &lt; 0.001; 95% CI 1.912-7.442; aOR 3.772), whereas endoscopic drainage of pancreatic necrosis (p = 0.018; 95% CI 0.138-0.834; aOR 0.339) and enteral nutrition (p = 0.003; 95% CI 0.143-0.716; aOR 0.320) were found as protective factors. Organ failure, acute cholangitis, and upfront open surgical necrosectomy were the most significant predictors of mortality. Our study confirmed that, even in a subgroup of particularly ill patients such as those with IPN, upfront open surgery should be avoided as much as possible. Study protocol registered in ClinicalTrials.Gov (I.D. Number NCT04747990)

Hepatitis B virus infection in hemodialysis: recent discoveries

Controlling the spread of HBV infection in dialysis units has been one of the major advances in the treatment of patients with ESRD. The frequency of HBsAg carriers on maintenance dialysis is low in developed countries. However, prevalence and incidence rates of HBV infection among dialysis patients in less developed countries remain very high. Patients on maintenance dialysis and chronic HBsAg carriage are typically anicteric and rarely develop symptoms of hepatitis. The relationship between HBsAg and aminotransferase activity in dialysis population is stronger than has so far been recognized. HBsAg positivity is significantly associated with hepatocellular damage in dialysis patients. Aminotransferase activity in dialysis population is usually depressed; this hampers the recognition of HBV-related liver disease among dialysis patients by biochemical tests. The HBV viral load in HBsAg positive patients is low and stable over time; the low mortality attributable to liver disease in dialysis patients could be related to this finding. Numerous assays for detecting HBV DNA in serum of dialysis patients are available; they should not be used for purposes of routine screening within dialysis units. Continued implementation of the long-recommended infection control procedures against HBV within dialysis units is strongly suggested

L'epatite E in Dialisi

Abstract non disponibil

Transplantation of kidneys from HCV-positive donors: a safe strategy?

Hepatitis C Virus (HCV) infection is the most important cause of liver disease after renal transplantation (RT). The impact of HCV on patient and graft survival after RT remains controversial; however, the great majority of studies with large size and adequate follow-up have shown the detrimental impact of HCV on long-term patient and graft survival after RT. The use of kidneys from anti-HCV positive donors could help decrease the continuing disparity between the number of patients on the transplant waiting list and the number of patients receiving a transplant each year. Single-center experiences have suggested transplanting kidneys from anti-HCV positive donors only in anti-HCV positive dialysis patients. Such practice has not demonstrated any adverse effect on the short-term patient survival; the waiting times for RT were shortened. A better alternative seems to be a policy of transplanting kidneys from anti-HCV positive donors only in HCV RNA positive recipients. This requires HCV RNA testing of all anti-HCV positive dialysis patients awaiting RT. Matching donors and recipients for HCV genotype has been suggested; however, the assessment of donor HCV genotype is currently hampered by time constraints. Recent evidence based on large data base demonstrated that RT recipients of HCV-positive donors are at independent increased risk of mortality; unadjusted 3-year patient survival was 85% versus 93% (P=0.01) in all recipients of donor HCV-positive and HCV-negative kidneys, respectively. This was observed in all recipient subgroups including elderly and HCV-positive recipients. In the near future, rapid nucleic acid testing (NAT) of donors and recipients will allow the assessment of the HCV viremic status in order to maximize organ use. With appropriate informed consent, use of a renal graft from an HCV positive donor may be offered to an HCV infected recipient. Additional studies are needed to clarify the link between donor HCV-positive kidneys and patient mortality

Kinetics of hepatitis C virus load during hemodialysis: novel perspectives

Hepatitis C virus (HCV) infection remains frequent among dialysis patients. The relationship between viral load and liver disease progression is currently a matter of debate; however, low HCV viral load (HCV RNA) is a well established predictor of successful antiviral treatment. Dialysis patients have immune compromise due to uremia; in spite of this, HCV viral load is not high and does not increase over time. A number of studies support the notion that this dynamics of HCV viral load may be related to lowering of HCV RNA titers during the HD procedure. It has been suggested that the intradialytic reduction of HCV is membrane-dependent; polysulphone (PS) and hemophan membranes appear more effective. Various mechanisms have been mentioned to explain the reduction of HCV RNA during hemodialysis (HD): adsorption of HCV onto dialysis membrane, HCV escape into spent dialysate, destruction of HCV particles or increased interferon (IFN) activity. Several investigators have noted that, at the end of the HD procedure, a virological rebound of HCV viremia occurs. Some suggestions have been made to minimize the risk for HCV escape into spent dialysate. However, controversial evidence on these issues exists: the HCV RNA reduction during HD procedures has not been seen in some studies and the relationship between the intra-dialytic reduction of HCV RNA titers and dialysis membrane is not completely understood. It is evident that additional long-term longitudinal studies by serial viral load estimations are needed to better define the dynamics of HCV viral load in the dialysis population

Management of hepatitis B after renal transplantation: an update

Hepatitis B Virus (HBV) infection remains an important cause of liver disease in renal transplant (RT) recipients and the outcome of HBV infected RT recipients is less favorable than that of noninfected RT recipients. The concern about graft loss induced by interferon (IFN) therapy precludes its use; lamivudine, a second-generation analog, has been recently approved for the treatment of hepatitis B and promises to be highly effective in this setting. Several uncontrolled trials have reported a high rate of biochemical (ranging between 80% and 100%) and virological (ranging between 67% and 100%) response in RT recipients, comparable to immunocompetent patients. Lamivudine has an excellent safety profile in RT recipients. However, the emergence of viral mutations leading to resistance has been reported during lamivudine therapy in RT recipients. In addition, numerous issues remain to be clarified about lamivudine use after RT including the management of viral resistance, the role of HBV genotypes in the response to lamivudine, and the duration of therapy and response. The combination of newer nucleoside analogues with lamivudine, analogous to HIV, may further improve the efficacy of antiviral therapy against HBV after RT