Auxilin is a novel susceptibility gene for congenital heart block which directly impacts fetal heart function

Objective: Neonatal lupus erythematosus (NLE) may develop after transplacental transfer of maternal autoantibodies with cardiac manifestations (congenital heart block, CHB) including atrioventricular block, atrial and ventricular arrhythmias, and cardiomyopathies. The association with anti-Ro/SSA antibodies is well established, but a recurrence rate of only 12%–16% despite persisting maternal autoantibodies suggests that additional factors are required for CHB development. Here, we identify fetal genetic variants conferring risk of CHB and elucidate their effects on cardiac function. Methods: A genome-wide association study was performed in families with at least one case of CHB. Gene expression was analysed by microarrays, RNA sequencing and PCR and protein expression by western blot, immunohistochemistry, immunofluorescence and flow cytometry. Calcium regulation and connectivity were analysed in primary cardiomyocytes and cells induced from pleuripotent stem cells. Fetal heart performance was analysed by Doppler/echocardiography. Results: We identified DNAJC6 as a novel fetal susceptibility gene, with decreased cardiac expression of DNAJC6 associated with the disease risk genotype. We further demonstrate that fetal cardiomyocytes deficient in auxilin, the protein encoded by DNAJC6, have abnormal connectivity and Ca2+ homoeostasis in culture, as well as decreased cell surface expression of the Cav1.3 calcium channel. Doppler echocardiography of auxilin-deficient fetal mice revealed cardiac NLE abnormalities in utero, including abnormal heart rhythm with atrial and ventricular ectopias, as well as a prolonged atrioventricular time intervals. Conclusions: Our study identifies auxilin as the first genetic susceptibility factor in NLE modulating cardiac function, opening new avenues for the development of screening and therapeutic strategies in CHB.publishedVersio

Activated allogeneic donor-derived marrow-infiltrating lymphocytes display tumor specificity and exhibit in vitro antitumor activity

A major issue in allogeneic hematopoietic stem cell transplantation (alloHSCT) is the difficulty of dealing with disease relapse after the transplant. The current standard approach is the use of donor lymphocyte infusions which have documented efficacy but are associated with significant mortality and relevant adverse events related to graft-versus-host disease (GvHD). Another recent approach is the development of adoptive T-cell therapies, but its major limitation is the limited availability of T-cells that are tumor-specific and display a broad antigenic repertoire. We show here that allogeneic donor-derived marrow-infiltrating lymphocytes (alloMILs) obtained from 9 patients who underwent alloHSCT with post-transplant cyclophosphamide and polyclonally stimulated with antiCD3/CD28 beads were reproducibly expanded and activated. Phenotypic characterization of activated alloMIL subpopulations revealed that the prevalent T-cell subtype exhibited a central memory phenotype in both CD4+ and CD8+ compartments. Activated alloMILs specifically recognized tumor cells and displayed in vitro antitumor activity. Selective expansion of small alloMIL subsets specific for different tumor antigens led to heightened tumor specificity and maintenance of a broad antigenic repertoire. Furthermore, activated alloMILs from all patients effectively targeted third-party allogeneic antigens, but did not show any reactivity towards autologous antigens presented in an HLA-dependent fashion, thereby suggesting a possible reduction in the risk of GvHD development. Collectively, these results underscore the intrinsic polyclonal tumor-specificity of activated alloMILs and describe a novel approach for the generation of tumor-specific T-cells that are suitable for adoptive immunotherapy of hematological malignancies relapsed after alloHSCT and that may significantly reduce the toxicities associated with current standard approaches

Determining Risk Factors for the Development of Temporomandibular Disorders during Orthodontic Treatment

Temporomandibular disorders (TMD) represent a complex disease with a multifactorial etiology. Despite several studies on the subject, a causal relationship between orthodontic treatment and different forms of TMD has not been established. The aim of this study was to analyze the effect of orthodontic treatment on two aspects of TMD: myofascial pain and disc displacement. This retrospective cohort study followed 224 orthodontic adult patients at three points in time: before treatment (T0), immediately after treatment (T1), and one year after treatment (T2). Disc displacement and myofascial pain were evaluated through a clinical assessment and with a semi-structured interview, along with headache, neck, and shoulder pain parameters and behavioral and somatic accompanying symptoms. Multivariate logistic regression was used to identify risk factors that could influence the development of TMD in these patients. There was a non-significant increase in disc displacement during orthodontic treatment, which mostly resolved after completion of treatment. Myofascial pain scores worsened during treatment, but improved when compared with the baseline once treatment was complete (T0 = 51.3%, T1 = 64.6%, T2 = 44.9%). Female gender (aOR = 1.9, CI 95%, 1.23–2.36), the presence of somatic symptoms (aOR = 3.6, CI 95%, 2.01–5.84), and symptoms of anxiety or depression (aOR = 2.2, CI 95%, 1.14–4.51) were significant risk factors associated with the development of TMD. There is a low and not significant risk of TMD development during orthodontic treatment. When TMD occurred, they resolved within 1 year of the end of treatment

Impacted and transmigrant mandibular canines: a systematic review

This study systematically reviewed the current literature on impacted and transmigrant lower canines, analyzing 1) current data of incidence, 2) updated research on etiology, 3) interceptive and active treatment strategies

Dental arch response to Haas-type rapid maxillary expansion anchored to deciduous vs permanent molars: A multicentric randomized controlled trial

Objective: To assess maxilla and mandibular arch widths' response to Haas-type rapid maxillary expansion (RME) anchored to deciduous vs permanent molars on children with unilateral posterior crossbite. Materials and Methods: Seventy patients with unilateral posterior crossbite recruited at the Universities of Genova, Siena, and Insubria (Varese) were randomly located into GrE (RME on second deciduous molars) or Gr6 (RME on first permanent molars) and compared. Results: Upper intermolar distance and permanent molar angulation increased significantly in Gr6 vs GrE at T1. Upper intercanine distance increased significantly in GrE vs Gr6 at T1 and T2. GrE showed significant increases for upper intermolar and upper intercanine widths. Gr6 showed statistically significant increases for upper intermolar widths, for upper and lower intercanine widths, and for increases of angulation of upper and lower permanent molars. Conclusions: GrE showed reduced molar angulation increases at T1 and reduced molar angulation decreases at T2 when compared with Gr6. At T2, the net increase of the upper intercanine distance in GrE was still significant compared with Gr6, indicating a more stable expansion in the anterior area

Non-COVID-19 patients in times of pandemic: Emergency department visits, hospitalizations and cause-specific mortality in Northern Italy

The COVID-19 pandemic forced healthcare services organization to adjust to mutating healthcare needs. Not exhaustive data are available on the consequences of this on non-COVID-19 patients. The aim of this study was to assess the impact of the pandemic on non-COVID-19 patients living in a one-million inhabitants\u2019 area in Northern Italy (Bologna Metropolitan Area-BMA), analyzing time trends of Emergency Department (ED) visits, hospitalizations and mortality. We conducted a retrospective observational study using data extracted from BMA healthcare informative systems. Weekly trends of ED visits, hospitalizations, in- and out-of-hospital, all-cause and cause-specific mortality between December 1st, 2019 to May 31st, 2020, were compared with those of the same period of the previous year. Non-COVID-19 ED visits and hospitalizations showed a stable trend until the first Italian case of COVID-19 has been recorded, on February 19th, 2020, when they dropped simultaneously. The reduction of ED visits was observed in all age groups and across all severity and diagnosis groups. In the lockdown period a significant increase was found in overall out-of-hospital mortality (43.2%) and cause-specific out-of-hospital mortality related to neoplasms (76.7%), endocrine, nutritional and metabolic (79.5%) as well as cardiovascular (32.7%) diseases. The pandemic caused a sudden drop of ED visits and hospitalizations of non-COVID-19 patients during the lockdown period, and a concurrent increase in out-of-hospital mortality mainly driven by deaths for neoplasms, cardiovascular and endocrine diseases. As recurrencies of the COVID-19 pandemic are underway, the scenario described in this study might be useful to understand both the population reaction and the healthcare system response at the early phases of the pandemic in terms of reduced demand of care and systems capability in intercepting it

Congenital heart block maternal sera autoantibodies target an extracellular epitope on the α1G T-type calcium channel in human fetal hearts.

Congenital heart block (CHB) is a transplacentally acquired autoimmune disease associated with anti-Ro/SSA and anti-La/SSB maternal autoantibodies and is characterized primarily by atrioventricular (AV) block of the fetal heart. This study aims to investigate whether the T-type calcium channel subunit α1G may be a fetal target of maternal sera autoantibodies in CHB.We demonstrate differential mRNA expression of the T-type calcium channel CACNA1G (α1G gene) in the AV junction of human fetal hearts compared to the apex (18-22.6 weeks gestation). Using human fetal hearts (20-22 wks gestation), our immunoprecipitation (IP), Western blot analysis and immunofluorescence (IF) staining results, taken together, demonstrate accessibility of the α1G epitope on the surfaces of cardiomyocytes as well as reactivity of maternal serum from CHB affected pregnancies to the α1G protein. By ELISA we demonstrated maternal sera reactivity to α1G was significantly higher in CHB maternal sera compared to controls, and reactivity was epitope mapped to a peptide designated as p305 (corresponding to aa305-319 of the extracellular loop linking transmembrane segments S5-S6 in α1G repeat I). Maternal sera from CHB affected pregnancies also reacted more weakly to the homologous region (7/15 amino acids conserved) of the α1H channel. Electrophysiology experiments with single-cell patch-clamp also demonstrated effects of CHB maternal sera on T-type current in mouse sinoatrial node (SAN) cells.Taken together, these results indicate that CHB maternal sera antibodies readily target an extracellular epitope of α1G T-type calcium channels in human fetal cardiomyocytes. CHB maternal sera also show reactivity for α1H suggesting that autoantibodies can target multiple fetal targets