Early Recovery of Aphasia through Thrombolysis: The Significance of Spontaneous Speech

Aphasia in one of the most devastating stroke-related consequences for social interaction and daily activities. Aphasia recovery in acute stroke depends on the degree of reperfusion after thrombolysis or thrombectomy. As aphasia assessment tests are often time-consuming for patients with acute troke, physicians have been developing rapid and simple tests. The aim of our study is to evaluate the improvement of language functions in the earliest stage in patients treated with thrombolysis and in nontreated patients using our rapid screening tes

A serine protease secreted from Bacillus subtilis cleaves human plasma transthyretin to generate an amyloidogenic fragment

Aggregation of human wild-type transthyretin (hTTR), a homo-tetrameric plasma protein, leads to acquired senile systemic amyloidosis (SSA), recently recognised as a major cause of cardiomyopathies in 1-3% older adults. Fragmented hTTR is the standard composition of amyloid deposits in SSA, but the protease(s) responsible for amyloidogenic fragments generation in vivo is(are) still elusive. Here, we show that subtilisin secreted from Bacillus subtilis, a gut microbiota commensal bacterium, translocates across a simulated intestinal epithelium and cleaves hTTR both in solution and human plasma, generating the amyloidogenic fragment hTTR(59-127), which is also found in SSA amyloids in vivo. To the best of our knowledge, these findings highlight a novel pathogenic mechanism for SSA whereby increased permeability of the gut mucosa, as often occurs in elderly people, allows subtilisin (and perhaps other yet unidentified bacterial proteases) to reach the bloodstream and trigger generation of hTTR fragments, acting as seeding nuclei for preferential amyloid fibrils deposition in the heart. Peterle et al. show that a subtilisin like serine protease secreted from gut microbiota Bacillus subtilis cleaves the wild-type human transthyretin (hTTR) to generate an amyloidogenic peptide. High propensity of the hTTR fragment to form pathogenic protein aggregates implicates the serine protease in the pathogenesis of acquired senile systemic amyloidosis

Comparison of different sets of immunological tests to identify treatable immunodeficiencies in adult bronchiectasis patients

BACKGROUND: The reported prevalence of immunodeficiencies in bronchiectasis patients is variable depending on the frequency and extent of immunological tests performed. European Respiratory Society guidelines recommend a minimum bundle of tests. Broadening the spectrum of immunological tests could increase the number of patients diagnosed with an immunodeficiency and those who could receive specific therapy. The primary objective of the present study was to assess the performance of different sets of immunological tests in diagnosing any, primary, secondary or treatable immunodeficiencies in adults with bronchiectasis. METHODS: An observational, cross-sectional study was conducted at the Bronchiectasis Program of the Policlinico University Hospital in Milan, Italy, from September 2016 to June 2019. Adult outpatients with a clinical and radiological diagnosis of bronchiectasis underwent the same immunological screening during the first visit when clinically stable consisting of: complete blood count; immunoglobulin (Ig) subclass tests for IgA, IgG, IgM and IgG; total IgE; lymphocyte subsets; and HIV antibodies. The primary endpoint was the prevalence of patients with any immunodeficiencies using five different sets of immunological tests. RESULTS: A total of 401 bronchiectasis patients underwent the immunological screening. A significantly different prevalence of bronchiectasis patients diagnosed with any, primary or secondary immunodeficiencies was found across different bundles. 44.6% of bronchiectasis patients had a diagnosis of immunodeficiency when IgG subclasses and lymphocyte subsets were added to the minimum bundle suggested by the guidelines. CONCLUSION: A four-fold increase in the diagnosis of immunodeficiencies can be found in adults with bronchiectasis when IgG subclasses and lymphocyte subsets are added to the bundle of tests recommended by guidelines

The Soluble Recombinant Neisseria meningitidis Adhesin NadAΔ351–405 Stimulates Human Monocytes by Binding to Extracellular Hsp90

The adhesin NadA favors cell adhesion/invasion by hypervirulent Neisseria meningitidis B (MenB). Its recombinant form NadAΔ351–405, devoid of the outer membrane domain, is an immunogenic candidate for an anti-MenB vaccine able to stimulate monocytes, macrophages and dendritic cells. In this study we investigated the molecular mechanism of NadAΔ351–405 cellular effects in monocytes. We show that NadAΔ351–405 (against which we obtained polyclonal antibodies in rabbits), binds to hsp90, but not to other extracellular homologous heat shock proteins grp94 and hsp70, in vitro and on the surface of monocytes, in a temperature dependent way. Pre-incubation of monocytes with the MenB soluble adhesin interfered with the binding of anti-hsp90 and anti-hsp70 antibodies to hsp90 and hsp70 at 37°C, a condition in which specific cell-binding occurs, but not at 0°C, a condition in which specific cell-binding is very diminished. Conversely, pre-incubation of monocytes with anti-hsp90 and anti-hsp70 antibodies did not affected NadAΔ351–405 cell binding in any temperature condition, indicating that it associates to another receptor on their plasma membrane and then laterally diffuses to encounter hsp90. Consistently, polymixin B interfered with NadAΔ351–405 /hsp90 association, abrogated the decrease of anti-hsp90 antibodies binding to the cell surface due to NadAΔ351–405 and inhibited adhesin-induced cytokine/chemokine secretion without affecting monocyte-adhesin binding. Co-stimulation of monocytes with anti-hsp90 antibodies and NadAΔ351–405 determined a stronger but polymixin B insensitive cell activation. This indicated that the formation of a recombinant NadA/hsp90/hsp70 complex, although essential for full monocyte stimulation, can be replaced by anti-hsp90 antibody/hsp90 binding. Finally, the activation of monocytes by NadAΔ351–405 alone or in the presence of anti-hsp90 antibodies were both inhibited by neutralizing anti-TLR4 antibodies, but not by anti-TLR2 antibodies. We propose that hsp90-dependent recruitment into an hsp90/hsp70/TLR4 transducing signal complex is necessary for the immune-stimulating activity of NadAΔ351–405 anti-MenB vaccine candidate

The T.O.S.C.A. Project: Research, Education and Care

Despite recent and exponential improvements in diagnostic- therapeutic pathways, an existing “GAP” has been revealed between the “real world care” and the “optimal care” of patients with chronic heart failure (CHF). We present the T.O.S.CA. Project (Trattamento Ormonale dello Scompenso CArdiaco), an Italian multicenter initiative involving different health care professionals and services aiming to explore the CHF “metabolic pathophysiological model” and to improve the quality of care of HF patients through research and continuing medical education

Differential effects of heparin and glucose on structural conformation of human alpha1-antitrypsin: evidence for a heparin-induced cleaved form of the inhibitor.

alpha1 Antitrypsin (alpha1AT) is the archetypal member of the serpin superfamily. Current knowledge of its inhibitory mechanism does not provide for any heparin-induced enhancement of serine proteinase inhibition. Since previous results have shown that an apparently altered alpha1AT form may be purified from the plasma of insulin-dependent diabetics by means of heparin-affinity chromatography, in the present work the possibility was tested that heparin at various concentrations modifies the structural conformation and function of human alpha1AT in the absence and presence of glucose, used at concentrations of 15 mM to mimick mild hyperglycemic conditions. Heparin was observed to bind strongly to alpha1AT, causing maximal enhancement of tryptophan fluorescence emission at 50 microg/ml, mostly in the presence of glucose. Circular dichroism spectra revealed that heparin with glucose caused the most relaxed, ordered structure of the inhibitor with increased heat stability. Modification in conformation was accompanied by loss of inhibitory activity, as demonstrated by the inability of alpha1AT to block bovine trypsin in the specific assay and by alterations of its immunological properties. However, despite inactivation, in the presence of heparin-both with and without glucose-alpha1AT was still able to bind trypsin, as revealed by inhibitor-to-proteinase complexes visible in both SDS- and nondenaturing electrophoreses. These complexes showed the same feature regardless of trypsin concentration and differed from those formed at a molar excess of the inhibitor in the absence of heparin, since they underwent rapid, intense fragmentation accompanied by complete loss of the secondary structure of the inhibitor. Even in the absence of trypsin, cleavage of alpha1AT was also observed to occur at both Val321- and Glu344- in the primary sequence of the inhibitor in the presence of 50 microg/ml heparin, with and without glucose. These results suggest that heparin binding to alpha1AT causes profound structural changes in the molecule, involving both the expulsion of the reactive site out of the molecule plane and a relaxed, heat-stable form of the inhibitor, rendered a substrate for the proteinase. Although glucose apparently does not affect alpha1AT functioning, it does enhance the effects of heparin on the alpha1AT structure. The possibility is discussed that, while heparin and glucose binding occurs at different sites on alpha1AT, glucose favors heparin binding by inducing a partially relaxed form in the inhibitor. Differences in structure and charge between the two substances account for both different individual effects on alpha1AT and the predominance of the effects of heparin

Effects of tDCS on reward responsiveness and valuation in Parkinson's patients with impulse control disorders

: Parkinson's disease (PD) patients with impulse control disorders (ICD) frequently report hypersensitivity to rewards. However, a few studies have explored the effectiveness of modulation techniques on symptoms experienced by these patients. In this study, we assessed the effect of anodal tDCS over the DLPFC on reward responsiveness and valuation in PD patients with ICD. 43 participants (15 PD patients with ICD, 13 PD without ICD, and 15 healthy matched controls) were asked to perform a reward-craving test employing both explicit (self-ratings of liking and wanting) and implicit (heart rate and skin conductance response) measures, as well as two temporal discounting tasks with food and money rewards. Each participant performed the experimental tasks during active anodal tDCS of the left dorsolateral prefrontal cortex (DLPFC), anodal tDCS of the primary motor cortex (M1), and sham tDCS. Results showed increased wanting and a steeper temporal discounting of rewards in PD with ICD compared to the other groups. Moreover, we found that PD without ICD exhibit reduced liking for rewards. tDCS results capable to modulate the altered intensity of PD patients' liking, but not wanting and temporal discounting of rewards in PD patients with ICD. These findings confirm that alterations in reward responsiveness and valuation are characteristics of impulse control disorders in patients with PD but suggest that anodal tDCS over the left DLPFC is not capable to influence these processes. At the same time, they provide new insight into affective experience of rewards in PD