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Hemophagocytic lymphohistiocytosis (HLH) is a potentially life-threatening entity resulting from the dysregulated activation of T-lymphocytes. Secondary HLH (sHLH) complicates various medical conditions. Similar to 2009 H1N1 influenza, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection appears to trigger a cytokine storm in a subset of individuals. The patient of 64 years old presented himself in the emergency department with cough and fever, and was subjected to the nasopharyngeal swab and tested positive for SARS-CoV-2, as expected from epidemiological data. Hospitalized in cohort isolation, he initially presented a multi-organ impairment and specifically a worsening of his renal function, initial coagulopathy, lymphopenia modest thrombocytopenia and systemic inflammatory commitment. During the course of the 6th day, a picture of a probable cytokine storm and clear multiple organ failure (MOF) was evident. Therefore a clinical diagnosis was made according to HScore criteria of secondary hemophagocytic lymphohistiocytosis. We started high-dose steroid therapy (dexamethasone 8 mg bid). And after 36 h, the patient was significantly improved: alert, oriented, weaned from non-invasive ventilation (NIV) and stable blood chemistry. In conclusion, in coronavirus disease 2019 (COVID-19) positive patients with evidence of cytokine storm and acute respiratory distress syndrome (ARDS), the levels of blood triglycerides and ferritin are useful and often decisive to comfort the diagnosis of sHLH if cytopenia of one or more lines is revealed.

Background: Physical effort is one of the natural stimuli capable of triggering airway obstruction in asthmatics, the so called exercise-induced bronchoconstriction in asthma (EIBa). This study was performed in subjects with mild persistent asthma suspected for EIBa, aiming to find predictors among functional parameters at rest and during exercise for developing EIBa. Methods: In 20 subjects with mild asthma in stable conditions who reported respiratory symptoms on exertion in the past, measurements of baseline functional respiratory parameters and airways responsiveness by a methacholine challenge were obtained on the first day of the study after an adequate pharmacological washout. The day after, a maximal symptom-limited incremental cardiopulmonary exercise test (CPExT) was performed, with subsequent, repeated maneuvers of maximal full forced expiration to monitor the FEV1 change at 1,3,5,7,10 and 15 minutes after the end of exercise for diagnosing EIBa.Results: 19 subjects aged 27±5 years completed the two-days protocol. No functional parameters both at rest and during effort were useful to predict EIBa after stopping exercise that actually occurred in 12 individuals. In contrast to asthmatics without EIBa, in those with EIBa, however, mean Inspiratory Capacity (IC) did not increase with increasing ventilatory requirements during CPExT because 6 of them (50%) displayed dynamic pulmonary hyperinflation (DH) throughout the exercise, as documented by the progressive increase of end-expiratory lung volume. This subgroup of asthmatics with EIBa who in turn showed earlier and greater post-exercise FEV1 fall had significantly lower forced mean expiratory flow between 25% and 75% of forced vital capacity (FEF25-75%) at rest (p20FEV1 (pConclusions No functional respiratory parameters either at rest or during the effort seem to predict EIBa in mild asthmatics after maximal exercise test. In those with EIBa, however, a subgroup developed DH during exercise, and this was associated with baseline reduced forced expiratory flow-rates at lower lung volumes and higher airway hyperresponsiveness, suggesting a prominent small airways impairment.

Background: Coronavirus disease 2019 (COVID-19) may predispose patients to venous thromboembolism (VTE) due to inflammation, hypoxia, immobilization, and diffuse intravascular coagulation, despite standard thrombopropylaxis. Our retrospective study reports the incidence of pulmonary embolism (PE) in patients with COVID-19 and severe respiratory failure(SRF) treated with intermediate to full-dose enoxaparin. .Methods: This retrospective case series analysed data from patients with COVID-19 pneumonia and severe respiratory failure (SRF) admitted to our Respiratory Intensive Care Unit (RICU) between February 27 and April 20, 2020 for non-invasive positive-pressure ventilation. All patients received at least intermediate-dose enoxaparin (40 mg twice daily). If PE was suspected or diagnosed, patients were treated with full-dose enoxaparin (1 mg/kg twice daily). Computed tomography pulmonary angiography (CTPA) was used to detect PE in patients with elevated D-dimer levels (> 3000 ng/mL) and/or other clinical indicators, including sudden worsening of cardiopulmonary status.Results: Ninety-two patients (71 males, 21 females; mean age 58 ± 11 years) with COVID-19 pneumonia and SRF (mean arterial oxygen partial pressure/fractional inspired oxygen [PaO2/FiO2] of 143 ± 45 mm Hg) were admitted to our RICU. Twenty-two patients underwent CTPA (24%), with PEs detected in 11 (12%). Mean PaO2/FiO2 and mean D-dimer levels did not significantly differ between patients with or without PE. Eleven patients (12%) died in the hospital, with a mean age of 70 ± 11 years for deceased patients and 56 ± 11 years for surviving patients (p < 0.0001).Conclusions: PE was diagnosed in 12% of patients despite intermediate to full-dose enoxaparin treatment. However the incidence of PE in our patients was lower than that previously reported. We hypothesize that this reduced PE incidence may have been secondary to the higher than prophylactic enoxaparin dose that was used.

Background: Physical effort is a natural stimulus capable of inducing acute airway obstruction in asthma: the so called exercise-induced asthma (EIA). Aim: Aim of the study was to identify the functional respiratory parameters able to predict the development of EIA in patients suffering of mild asthma. Methods: In 20 patients with mild asthma and suspected of previous EIA, after adequate drugs wash-out, baseline pulmonary function tests and non-specific bronchial stimulation with methacholine were performed. The next day they underwent a symptoms-limited incremental cardio-pulmonary exercise test (CPExT) with measurements of FEV1 at 1,3,5,7,10,15 minutes after the end of the exercise to establish the EIA development. Results: Only 19 patients (aged 27±5 yrs) completed the two-day protocol. Twelve of them experienced EIA. None of the baseline functional respiratory parameters measured at rest was able to predict EIA. In contrast with asthmatics without EIA, in those showing EIA the average Inspiratory Capacity (IC) did not increase during CPExT, because 6 of them (50%) displayed exertional dynamic pulmonary hyperinflation (DH), as documented by a progressive increase of end-expiratory lung volume. These asthmatics, who showed an earlier and larger drop in FEV1 at the end of the exercise, had lower FEF25-75% (%pred.) at rest (p Conclusions: Although this study could not identify predictors for EIA in patients with mild asthma, it demonstrated that asthmatics with EIA who developed DH during exercise, had lower resting FEF25-75% (% pred.) and greater airway hyper-responsiveness, suggesting a prevalent small airways impairment.

Background: The pandemic of new severe acute respiratory syndrome (SARS) due to coronavirus (CoV) 2 (SARS-CoV-2) has stressed the importance of effective diagnostic and prognostic biomarkers of clinical worsening and mortality. Epidemiological data showing a differential impact of SARS-CoV-2 infection on women and men have suggested a potential role for testosterone (T) in determining gender disparity in the SARS-CoV-2 clinical outcomes. Objectives: To estimate the association between T level and SARS-CoV-2 clinical outcomes (defined as conditions requiring transfer to higher or lower intensity of care or death) in a cohort of patients admitted in the respiratory intensive care unit (RICU). Materials and methods: A consecutive series of 31 male patients affected by SARS-CoV-2 pneumonia and recovered in the respiratory intensive care unit (RICU) of the “Carlo Poma” Hospital in Mantua were analyzed. Several biochemical risk factors (ie, blood count and leukocyte formula, C-reactive protein (CRP), procalcitonin (PCT), lactate dehydrogenase (LDH), ferritin, D-dimer, fibrinogen, interleukin 6 (IL-6)) as well as total testosterone (TT), calculated free T (cFT), sex hormone–binding globulin (SHBG), and luteinizing hormone (LH) were determined. Results: Lower TT and cFT were found in the transferred to ICU/deceased in RICU group vs groups of patients transferred to IM or maintained in the RICU in stable condition. Both TT and cFT showed a negative significant correlation with biochemical risk factors (ie, the neutrophil count, LDH, and PCT) but a positive association with the lymphocyte count. Likewise, TT was also negatively associated with CRP and ferritin levels. A steep increase in both ICU transfer and mortality risk was observed in men with TT&nbsp

p53 is the central regulator of cell fate following genotoxic stress and oncogene activation. Its activity is controlled by several posttranslational modifications. Originally defined as a critical layer of p53 regulation in human cell lines, p53 lysine methylation by Set7/9 (also called Setd7) was proposed to fulfill a similar function in vivo in the mouse, promoting p53 acetylation, stabilization, and activation upon DNA damage (Kurash et al., 2008). We tested the physiological relevance of this circuit in an independent Set7/9 knockout mouse strain. Deletion of Set7/9 had no effect on p53-dependent cell-cycle arrest or apoptosis following sublethal or lethal DNA damage induced by radiation or genotoxic agents. Set7/9 was also dispensable for p53 acetylation following irradiation. c-myc oncogene-induced apoptosis was also independent of Set7/9, and analysis of p53 target genes showed that Set7/9 is not required for the p53-dependent gene expression program. Our data indicate that Set7/9 is dispensable for p53 function in the mouse.

Ca2+-binding proteins containing EF-hands are important constituents of intracellular signaling pathways. Recently, three new members of the Neuronal Calcium Sensor subgroup have been cloned in humans. Calsenilin interacts with presenilins, DREAM is a calcium-regulated transcriptional repressor and KChIP3 binds and modulates A-type potassium channels. Here we describe the mouse full-length cDNA and the genomic locus, demonstrating that the three proteins are encoded by the same unique gene. Various mechanisms contribute to the coding potential of this locus. These include alternate translation starts in the first exon and alternative splicing yielding transcripts lacking the EF-hand domains. In situ hybridization, RT-PCR, and Northern blotting reveal nervous system-restricted expression largely coinciding with the distribution of the Kv4.2 α-subunit of potassium channels. The presence of transcripts in early embryonic stages suggests roles for the protein also during development.

PTX3 is a prototypic long pentraxin consisting of a C-terminal 203-amino acid pentraxin-like domain coupled with an N-terminal 178-amino acid unrelated portion. The present study was designed to characterize the structure and ligand binding properties of human PTX3, in comparison with the classical pentraxins C-reactive protein and serum amyloid P component. Sequencing of Chinese hamster ovary cell-expressed PTX3 revealed that the mature secreted protein starts at residue 18 (Glu). Lectin binding and treatment withN-glycosidase F showed that PTX3 isN-glycosylated, sugars accounting for 5 kDa of the monomer mass (45 kDa). Circular dichroism analysis indicated that the protein consists predominantly of β-sheets with a minor α-helical component. While in gel filtration the protein is eluted with a molecular mass of ≅900 kDa, gel electrophoresis using nondenaturing, nonreducing conditions revealed that PTX3 forms multimers predominantly of 440 kDa apparent molecular mass, corresponding to decamers, and that disulfide bonds are required for multimer formation. The ligand binding properties of PTX3 were then examined. As predicted based on modeling, inductive coupled plasma/atomic emission spectroscopy showed that PTX3 does not have coordinated Ca2+. Unlike the classical pentraxins CRP and SAP, PTX3 did not bind phosphoethanolamine, phosphocholine, or high pyruvate agarose. PTX3 in solution, bound to immobilized C1q, but not C1s, and, reciprocally, C1q bound to immobilized PTX3. Binding of PTX3 to C1q is specific and saturable with a K d 7.4 × 10−8 m as determined by solid phase binding assay. The Chinese hamster ovary cell-expressed pentraxin domain bound C1q when multimerized. Thus, as predicted on the basis of computer modeling, the prototypic long pentraxin PTX3 forms multimers, which differ from those formed by classical pentraxins in terms of protomer composition and requirement for disulfide bonds, and does not recognize CRP/SAP ligands. The capacity to bind C1q, mediated by the pentraxin domain, is consistent with the view that PTX3, produced in tissues by endothelial cells or macrophages in response to interleukin-1 and tumor necrosis factor, may act as a local regulator of innate immunity.

SummaryJMJD3 (KDM6B) antagonizes Polycomb silencing by demethylating lysine 27 on histone H3. The interplay of methyltransferases and demethylases at this residue is thought to underlie critical cell fate transitions, and the dynamics of H3K27me3 during neurogenesis posited for JMJD3 a critical role in the acquisition of neural fate. Despite evidence of its involvement in early neural commitment, however, its role in the emergence and maturation of the mammalian CNS remains unknown. Here, we inactivated Jmjd3 in the mouse and found that its loss causes perinatal lethality with the complete and selective disruption of the pre-Bötzinger complex (PBC), the pacemaker of the respiratory rhythm generator. Through genetic and electrophysiological approaches, we show that the enzymatic activity of JMJD3 is selectively required for the maintenance of the PBC and controls critical regulators of PBC activity, uncovering an unanticipated role of this enzyme in the late structuring and function of neuronal networks.

Work of the last decade has proven the ‘one gene– one product–one function’ hypothesis an oversimplification. To further unravel the emerging ‘one gene–multiple products–even more functions’ concept, new methods (such as subtle knock-in and tightly regulated conditional mutations) for the analysis of gene function in health and disease are required. Another class of improvements (such as tetraploid fusion and cassette exchange) addresses the efficiency with which targeted mutant strains can be generated. Recombinase-mediated cassette exchange (RMCE), which in theory is well suited for the rapid generation of multiple alleles of a given locus, is hampered by its low efficiency in the absence of selection and, especially in vivo, by the promiscuity of the participating recombinase recognition sites. Here we present a novel approach which circumvents this problem by the use of two independent recombinase systems. The strategy, which uses loxP on one and FRT on the other side of the cassette together with a Cre/Flpe expression vector, prevents excisive events and results in higher rates of cassette integration without selection than previously described. This method has a huge potential for the generation of allelic series in embryonic stem cells and, importantly, in pre-implantation embryos in vivo.

The pcp2/L7 gene is characterized by its very cell type-specific expression restricted to cerebellar Purkinje cells and retinal bipolar neurons. Although remarkable progress as to the biochemical properties of the encoded protein has been made, knowledge on its physiological functions remains sparse. While characterizing a pcp2-driven transgenic strain, we observed the presence of a longer, so far unknown, pcp2 transcript. Different from another recently discovered splice variant, ret-pcp2, expression of this novel transcript is observed in bipolar as well as cerebellar Purkinje cells of mid-postnatal mice. The protein encoded by our novel variant appears to be less efficient in binding to Gα subunits compared to the original L7/pcp2 protein and it is also less inhibitory with respect to GTPγ binding. Its expression in the eye appears to be independent from eye opening in postnatal mice.

Coronavirus disease (COVID-19) may predispose patients to pulmonary embolism (PE), despite standard thromboprophylaxis. Our retrospective study aimed to report the prevalence of PE in patients with COVID-19 and severe respiratory failure (SRF) treated with intermediate- to full-dose enoxaparin. We analyzed data from patients with COVID-19 pneumonia and SRF admitted to our Respiratory Intensive Care Unit (RICU) from February 27 to April 20, 2020. All patients received at least intermediate-dose enoxaparin (40 mg twice daily). Computed tomography pulmonary angiography (CTPA) was used to detect PE. Ninety-two patients with COVID-19 pneumonia and SRF were admitted to our RICU. Twenty-two patients underwent CTPA (24 %), 11 of whom had PEs (12%). We hypothesize that the enoxaparin treatment may be responsible for the lower prevalence of PE as compared to previous reports of similar patients, even if our report had several limitations, mainly the small sample size.