ANO3 as a Cause of Early‐Onset Chorea Combined with Dystonia: Illustration of Phenotypic Evolution

Here, we signpost a case of a childhood-onset chorea-dominant phenotype later evolved into a dystonia-dominant phenotype during adulthood, in a subject carrying a missense pathogenic variant (c.1528 G > A; p.Glu510Lys)1 in the anoctamin 3 protein-coding gene (ANO3, DYT24, OMIM 610110).

S-CMC-Lys protective effects on human respiratory cells during oxidative stress.

The mucoactive drug S-carbocysteine lysine salt monohydrate (S-CMC-Lys) stimulates glutathione (GSH) efflux from respiratory cells. Since GSH is one of the most important redox regulatory mechanisms, the aim of this study was to evaluate the S-CMC-Lys effects on GSH efflux and intracellular concentration during an oxidative stress induced by the hydroxyl radical (xOH). Experiments were performed on cultured human respiratory WI-26VA4 cells by means of patch-clamp experiments in whole-cell configuration and of fluorimetric analyses at confocal microscope. xOH exposure induced an irreversible inhibition of the GSH and chloride currents that was prevented if the cells were incubated with S-CMC-Lys. In this instance, the currents were inhibited by the specific blocker CFTR(inh)-172. CFT1-C2 cells, which lack a functional CFTR channel, were not responsive to S-CMC-Lys, but the stimulatory effect of the drug was restored in LCFSN-infected CFT1 cells, functionally corrected to express CFTR. Fluorimetric measurements performed on the S-CMC-Lys-incubated cells revealed a significant increase of the GSH concentration that was completely hindered after oxidative stress and abolished by CFTR(inh)-172. The cellular content of reactive oxygen species was significantly lower in the S-CMC-Lys-treated cells either before or after xOH exposure. As a conclusion, S-CMC-Lys could exert a protective function during oxidative stress, therefore preventing or reducing the ROS-mediated inflammatory response

Lactiplantibacillus plantarum monolayer enhanced bactericidal action of carvacrol: biofilm inhibition of viable foodborne pathogens and spoilage microorganisms

The prevalence of biofilm-associated microorganisms and the increasing use of ready-to-eat fresh products represent the current duality the food industry must address. Innovative and eco-friendly antibiofilm solutions and appropriate microbiological food control systems are urgently needed to improve food quality and safety. This study aimed to investigate the in vitro combined efficacy of carvacrol with a pre-formed biofilm monolayer of the probiotic Lactiplantibacillus plantarum DSM 20174. The antimicrobial activity of carvacrol against both planktonic and sessile cells of foodborne pathogens and spoilage microorganisms, alone or in the presence of the pre-formed biofilm of L. plantarum, was investigated by culture-based methods along with flow cytometry (FCM) to monitor cells' cultivability and viability. The synergistic action of carvacrol and the pre-formed biofilm of L. plantarum was evaluated in the 96-well plates. The results showed that L. plantarum pre-formed biofilm monolayer enhanced the antimicrobial effect of carvacrol determining a bactericidal action while the treatment alone induced the viable but not culturable (VBNC) cell state only. Furthermore, the great efficacy of the combined treatment allowed the application of a lower concentration of carvacrol (100 ppm) to achieve significant damage in cell viability. In conclusion, the incorporation of carvacrol into the L. plantarum pre-formed biofilm represents a promising alternative for an antimicrobial functionalized ready-to-eat packaging

Variants in ATP5F1B are associated with dominantly inherited dystonia

Nasca et al. identify a new candidate gene for dystonia, ATP5F1B, encoding a subunit of the mitochondrial ATP synthase (complex V). Likely pathogenic variants in ATP5F1B were associated with early-onset isolated dystonia in two independent families, both with an autosomal dominant mode of inheritance and incomplete penetrance. ATP5F1B is a subunit of the mitochondrial ATP synthase or complex V of the mitochondrial respiratory chain. Pathogenic variants in nuclear genes encoding assembly factors or structural subunits are associated with complex V deficiency, typically characterized by autosomal recessive inheritance and multisystem phenotypes. Movement disorders have been described in a subset of cases carrying autosomal dominant variants in structural subunits genes ATP5F1A and ATP5MC3. Here, we report the identification of two different ATP5F1B missense variants (c.1000A>C;p.Thr334Pro and c.1445T>C;p.Val482Ala) segregating with early-onset isolated dystonia in two families, both with autosomal dominant mode of inheritance and incomplete penetrance. Functional studies in mutant fibroblasts revealed no decrease of ATP5F1B protein amount but severe reduction of complex V activity and impaired mitochondrial membrane potential, suggesting a dominant-negative effect. In conclusion, our study describes a new candidate gene associated with isolated dystonia and confirms that heterozygous variants in genes encoding subunits of the mitochondrial ATP synthase may cause autosomal dominant isolated dystonia with incomplete penetrance, likely through a dominant-negative mechanism

Mutations in the Neuronal Vesicular SNARE VAMP2 Affect Synaptic Membrane Fusion and Impair Human Neurodevelopment

VAMP2 encodes the vesicular SNARE protein VAMP2 (also called synaptobrevin-2). Together with its partners syntaxin-1A and synaptosomal-associated protein 25 (SNAP25), VAMP2 mediates fusion of synaptic vesicles to release neurotransmitters. VAMP2 is essential for vesicular exocytosis and activity-dependent neurotransmitter release. Here, we report five heterozygous de novo mutations in VAMP2 in unrelated individuals presenting with a neurodevelopmental disorder characterized by axial hypotonia (which had been present since birth), intellectual disability, and autistic features. In total, we identified two single-amino-acid deletions and three non-synonymous variants affecting conserved residues within the C terminus of the VAMP2 SNARE motif. Affected individuals carrying de novo non-synonymous variants involving the C-terminal region presented a more severe phenotype with additional neurological features, including central visual impairment, hyperkinetic movement disorder, and epilepsy or electroencephalography abnormalities. Reconstituted fusion involving a lipid-mixing assay indicated impairment in vesicle fusion as one of the possible associated disease mechanisms. The genetic synaptopathy caused by VAMP2 de novo mutations highlights the key roles of this gene in human brain development and function

AI is a viable alternative to high throughput screening: a 318-target study

: High throughput screening (HTS) is routinely used to identify bioactive small molecules. This requires physical compounds, which limits coverage of accessible chemical space. Computational approaches combined with vast on-demand chemical libraries can access far greater chemical space, provided that the predictive accuracy is sufficient to identify useful molecules. Through the largest and most diverse virtual HTS campaign reported to date, comprising 318 individual projects, we demonstrate that our AtomNet® convolutional neural network successfully finds novel hits across every major therapeutic area and protein class. We address historical limitations of computational screening by demonstrating success for target proteins without known binders, high-quality X-ray crystal structures, or manual cherry-picking of compounds. We show that the molecules selected by the AtomNet® model are novel drug-like scaffolds rather than minor modifications to known bioactive compounds. Our empirical results suggest that computational methods can substantially replace HTS as the first step of small-molecule drug discovery

Entry, market structure and innovation in a history-friendly model of the evolution of the pharmaceutical industry

CAMBRIDGE UNIVERSITY PRES

Using Function Points to Measure and Estimate Real-Time and Embedded Software: Experiences and Guidelines

The developers of real-time and embedded software face \u2013just like the developers of other types of software\u2013 the problem of estimating the cost of development. To this end, the most widely used methods and tools require that the functional size of the program to be developed is measured. However, the functional size measurement methods available \u2013namely, Function Point Analysis and its evolutions\u2013 are traditionally considered not well suited for representing the functionality of real-time and embedded software. Actually, the problem is that the definition of Function Points and their counting rules make reference almost exclusively to traditional \u201cbusiness\u201d software. In this paper, the problem of applying FPA to embedded and real-time software is tackled. A set of hints and examples \u2013derived from industrial experience\u2013 are given, supporting the application of standard function point counting to real-time and embedded software. It is then shown that the obtained measures successfully supported the estimation of a set of programs in the avionics domain

Entry, Market Structure and Innovation in a History-Friendly Model of the Evolution of the Pharmaceutical Industry

CAMBRIDGE UNIVERSITY PRES