Characterization of a disease-associated mutation affecting a putative splicing regulatory element in intron 6b of the cystic fibrosis transmembrane conductance regulator (CFTR) gene

Cystic fibrosis (CF) is a common recessive disorder caused by >1600 mutations in the CF transmembrane conductance regulator (CFTR) gene. About 13% of CFTR mutations are classified as “splicing mutations,” but for almost 40% of these, their role in affecting the pre-mRNA splicing of the gene is not yet defined. In this work, we describe a new splicing mutation detected in three unrelated Italian CF patients. By DNA analyses and mRNA studies, we identified the c.1002–1110_1113delTAAG mutation localized in intron 6b of the CFTR gene. At the mRNA level, this mutation creates an aberrant inclusion of a sequence of 101 nucleotides between exons 6b and 7. This sequence corresponds to a portion of intron 6b and resembles a cryptic exon because it is characterized by an upstream ag and a downstream gt sequence, which are most probably recognized as 5′- and 3′-splice sites by the spliceosome. Through functional analysis of this splicing defect, we show that this mutation abolishes the interaction of the splicing regulatory protein heterogeneous nuclear ribonucleoprotein A2/B1 with an intronic splicing regulatory element and creates a new recognition motif for the SRp75 splicing factor, causing activation of the cryptic exon. Our results show that the c.1002–1110_1113delTAAG mutation creates a new intronic splicing regulatory element in intron 6b of the CFTR gene exclusively recognized by SRp75

Alexithymia for cardiologists: a clinical approach to the patient

Alexithymia literally meaning ‘no words for emotions’ is a term used in mental health settings to describe people who have difficulties in identifying and verbalizing their emotional states. There is evidence in the literature that this personality trait may influence negatively the illness behavior when an acute coronary event occurs. In fact, people with high alexithymia are more likely to experience wrong appraisal and interpretation of symptoms, and because of their difficulty in describing feelings to others, they can be poor in reporting symptoms at the first consultation with a physician. This behavioral pattern (alexithymic) may put patients with acute myocardial infarction at higher risk for delayed medical care. Here, we aim to present an overview of alexithymia from the perspective of the clinical cardiologist, with a focus on the definition, clinical recognition, and potential impact on cardiovascular health

Listeria monocytogenes in Five Sardinian Swine Slaughterhouses: Prevalence, Serotype, and Genotype Characterization

In a 3-year study (2008 to 2011) to estimate the prevalence and the contamination sources of Listeria monocytogenes in pork meat in Sardinia, Italy, 211 samples were collected from five Sardinian swine slaughterhouses: 171 samples from slaughtered pigs and 40 from the slaughterhouse environment. Fifty L. monocytogenes isolates were characterized by PCR-based serotyping, presence of virulence-associated genes, and pulsed-field gel electrophoresis restriction analysis. The overall prevalence of L. monocytogenes was 33% in swine carcasses, 7% in cecal material, 23% on meat contact surfaces, and 25% on noncontact surfaces. Only two serotypes were detected: 1/2c (78%) and 1/2a (22%). In all, based on the presence of virulence-associated genes, eight pathogenic profiles were detected. Only 42% of all isolates carried the full complement of virulence-associated genes and were allotted to profile 1. Six pulsed-field gel electrophoresis profiles persisted in the slaughterhouses; restriction profiles appeared to be specific to each plant

Vitamin E–loaded dialyzer resets PBMC-operated cytokine network in dialysis patients

Vitamin E–loaded dialyzer resets PBMC-operated cytokine network in dialysis patients.BackgroundIn hemodialysis patients the activity of stimulated Th1 lymphocytes is depressed, while Th2 cells are constitutively primed. Such phenomena may depend on monocyte activation and altered release of interleukin (IL)-12 and IL-18, which regulate Th cell differentiation. Reactive oxygen species (ROS) activate monocytes; therefore, a hemodialyzer with antioxidant activity would contrast ROS, prevent monocyte activation, reset IL-12 and IL-18 release, and restore Th1/Th2 balance.MethodsTen patients on regular dialysis treatment (RDT) with cellulosic membrane (CM) were shifted to vitamin E–coated dialyzer (VE). During treatment with CM and after 3, 6, and 12months of treatment with VE, peripheral blood mononuclear cells (PBMC) and purified CD4+ cells were isolated, and cultured, resting, mitogen-stimulated, and interferon γ (IFNγ), IL-4, IL-10, IL-12, and IL-18 release was measured. Vitamin E and A plasma levels and the effects of a single dialysis session on peripheral blood NO levels were assayed.ResultsThe constitutive release of IL-4 and IL-10 by CD4+ cells was abated significantly by treatment with VE (nadir -77.8% and -55.3%, respectively, at 12months). INFγ release by mitogen-stimulated CD4+ recovered with VE (zenith +501% at 12months). PBMC constitutive production of IL-12 and IL-18 was significantly reduced by VE (nadir at 12months -64.7% and -51.3%, respectively). VE increased plasma levels of vitamins E and A. NO plasma levels fell after a single dialysis treatment with VE (-17%, P < 0.05) in contrast with CU (+27.1%, P < 0.05).ConclusionThe network of cytokines released by monocytes and Th cells is reset toward normality by treatment with vitamin E–coated dialyzer

AIRE acetylation and deacetylation: effect on protein stability and transactivation activity

The AIRE protein plays a remarkable role as a regulator of central tolerance by controlling the promiscuous expression of tissue-specific antigens in thymic medullary epithelial cells. Defects in AIRE gene cause the autoimmune polyendocrinopathy- candidiasis-ectodermal dystrophy, a rare disease frequent in Iranian Jews, Finns, and Sardinian population. AIRE protein is primarily known as a transcriptional regulator and is capable of interacting with numerous proteins. The first characterized partner of AIRE is the ubiquitous transcription factor CREB-binding protein (CBP), which regulates DNA transcription through the acetylation and deacetylation of histones. More recently, the role of p300 in AIRE acetylation, which could influence the selection of AIRE activated genes, has been described. Results: In this study, we have precisely mapped, by mass spectrometry experiments, the sites of protein acetylation and, by mutagenesis assays, we have described a set of acetylated lysines as being crucial in influencing the subcellular localization of AIRE. Furthermore, we have also determined that the de-acetyltransferase enzymes HDAC1-2 are involved in the lysine de-acetylation of AIRE. Conclusions: On the basis of our results and those reported in literature, we propose a model in which lysines acetylation increases the stability of AIRE in the nucleus. In addition, we observed that the interaction of AIRE with deacetylases complexes inhibits its transcriptional activity and is probably responsible for the instability of AIRE, which becomes more susceptible to degradation in the proteasome

Quark mixing in the discrete dark matter model

We consider a model in which dark matter is stable as it is charged under a Z2 symmetry that is residual after an A4 flavour symmetry is broken. We consider the possibility to generate the quark masses by charging the quarks appropriately under A4. We find that it is possible to generate the CKM mixing matrix by an interplay of renormalisable and dimension-six operators. In this set-up, we predict the third neutrino mixing angle to be large and the dark matter relic density to be in the correct range. However, low energy observables - in particular meson-antimeson oscillations - strongly limit the available parameter space.Comment: 10 pages, 4 figures; version 3 as appeared in Nuclear Physics

650 Quality of life of patients with end stage heart failure treated with left ventricular assist devices

Abstract Aims Nowadays continuous flow left ventricular assist devices (LVAD) have become a reality for patients with end-stage heart failure (HF) who are failing maximal medical treatment, both eligible or not for heart transplantation. LVADs have demonstrated to improve functional capacity and clinical outcomes, including breathing and activity tolerance, but the impact on patients quality of life (QoL) is still a vexed and open question. Moreover, the device implantation actually requires significant life style changes, high motivation and adherence to treatment, both for patients and their caregivers. Patients must learn to live with the device and to interface with the controller and batteries system, adjusting everyday life's activities to the device presence. The purpose of our study is to evaluate how the implantation of HeartMate 3™ impact on patients-related QoL and clinical outcomes, compared with general population and other chronic diseases. Methods and results Eight patients (pts) with a diagnosis of end-stage HF were implanted with the HeartMate 3™ LVAD from May 2017 to October 2019 in the Cardiac Surgery Unit of Brotzu Hospital. During a follow-up visit (28.7 ± 11.9 months after surgery), the 7 pts presenting were assessed with two questionnaires, the SF-12 and EuroQoL-5D, in order to evaluate improvement of quality of life compared with clinical presentation. Afterwards we compared the average SF-12 total result with 28 controls from general population and with other important chronic diseases. The analysis of SF-12 questionnaires showed an average of 28.00 ± 8.98 (VN: 12–47) as total score, 11.7 ± 3.4 (VN: 6–20) as physical health component score and 16.2 ± 6.5 (VN: 6–27) as mental health component score. These findings, though within the range of 'normality', are closer to the lower scores and show the negative impact of LVAD in everyday patients-related quality of life. Comparing the NYHA functional class with these results, we found a statistically significant negative linear correlation for both total (−0.80, P = 0.03) and mental health component scores (−0.75, P = 0.049). Average total score of our pts significatively differs compared with 28 controls of general population (28.00 ± 8.98 vs. 38.64 ± 6.80, P = 0.014), as well as compared with other chronic diseases like Wilson's disease (P &lt; 0.001), celiac disease (P &lt; 0.001), obsessive-compulsive disorder (P &lt; 0.001), panic disorder (P &lt; 0.001), major depressive disorder (P = 0.009), multiple sclerosis (P &lt; 0.011), food disorders (P = 0.023), and carotid atherosclerosis (P = 0.049). The EuroQoL-5D questionnaire investigates five particular 'dimensions' of the subjectively perceived health-related quality of life (mobility, self-care, usual activities, pain/discomfort, and anxiety/depression) and shows similar. We found a poor average score at Visual Analogue Scale (58.5 ± 18.86) and a statistically significant negative linear correlation with NYHA functional class (−0.76, P = 0.046). The 42.86% referred no pain or discomfort and in the other four dimensions most of them reported 'moderate limitations', both physical or mental, especially for self-care domain (85.71%), as inevitable result of the device's size. Conclusions LVAD can improve clinical outcomes and functional capacity of carefully select pts with end-stage HF, but the complications encountered during mechanical support and the lifestyle changes required can affect negatively patients well-being. Many studies have shown that most patients experience significant improvement even in QoL's perception but for others the device have a negative impact on many aspects of normal daily living as well as emotional, mental and social functioning. Our study confirms this conflicting results: physical related-quality of life improves after LVAD implantation but emotional and psychological distress may persist, especially during long-term support, as a result of complications, co-morbidities and personal's attitudes, values and way of life

Gender differences and cardiometabolic risk. the importance of the risk factors

Metabolic syndrome (Mets) is a clinical condition characterized by a cluster of major risk factors for cardiovascular disease (CVD) and type 2 diabetes: proatherogenic dyslipidemia, elevated blood pressure, dysglycemia, and abdominal obesity. Each risk factor has an independent effect, but, when aggregated, they become synergistic, doubling the risk of developing cardiovascular diseases and causing a 1.5-fold increase in all-cause mortality. We will highlight gender differences in the epidemiology, etiology, pathophysiology, and clinical expression of the aforementioned Mets components. Moreover, we will discuss gender differences in new biochemical markers of metabolic syndrome and cardiovascular risk