The current coronavirus disease-2019 (COVID-19) pandemic is due to the novel coronavirus SARS-CoV-2. The scientific community has mounted a strong response by accelerating research and innovation, ...and has quickly set the foundation for understanding the molecular determinants of the disease for the development of targeted therapeutic interventions. The replication of the viral genome within the infected cells is a key stage of the SARS-CoV-2 life cycle. It is a complex process involving the action of several viral and host proteins in order to perform RNA polymerization, proofreading and final capping. This review provides an update of the structural and functional data on the key actors of the replicatory machinery of SARS-CoV-2, to fill the gaps in the currently available structural data, which is mainly obtained through homology modeling. Moreover, learning from similar viruses, we collect data from the literature to reconstruct the pattern of interactions among the protein actors of the SARS-CoV-2 RNA polymerase machinery. Here, an important role is played by co-factors such as Nsp8 and Nsp10, not only as allosteric activators but also as molecular connectors that hold the entire machinery together to enhance the efficiency of RNA replication.
Glycoconjugate vaccines have been effectively used in humans for about 40 years. The glycoconjugates have substituted plain polysaccharide vaccines that have many limitations, especially in infants. ...The covalent linking of protein to carbohydrates has allowed to overcome T-cell-dependent type-2 response of sugars. Glycoconjugates can show improved responses (over plain saccharides) also in elderly and immunocompromised (and depending on the endpoint also in immunocompetent adults), but infants represent the main target of these vaccines because of their unique immune system. Differently from the plain polysaccharide vaccines, the glycoconjugates are also able to induce Immunoglobulin G (IgG) response in infants.
Recently, vaccines containing conjugates directly expressed in Escherichia coli (bioconjugates) have been tested in the clinic against Shigella dysenteriae type 1, uropathogenic E. coli, and Streptococcus pneumoniae.
Here, we report an overall comparison of classical- and bioconjugate vaccines in terms of the structural properties and the immunological response elicited.
•Chemical conjugates are obtained by covalent coupling of polysaccharide to carrier protein.•Chemical conjugates are a very consolidated platform.•Bioconjugates are conjugates directly expressed in Escherichia coli.•Bioconjugates tested in clinic against Shigella dysenteriae, E.coli and S.pneumoniae.•Overall comparison of characteristics and immunological response of conjugate vaccines.
Within a principal‐agent model, the paper studies how hidden information affects incentives to invest in demand‐enhancing R&D of a firm competing in the product market. The analysis shows that, when ...the R&D outcome is private information of the innovating firm not only compared to its competitor, but also relative to its supplier, a contractual cost arises which neutralizes the standard strategic benefit of R&D and reduces the incentives to invest. Moreover, within this setting, more intense competition always stifles innovation.
The availability of standardized guidelines regarding the use of electronic fetal monitoring (EFM) in clinical practice has not effectively helped to solve the main drawbacks of fetal heart rate ...(FHR) surveillance methodology, which still presents inter- and intra-observer variability as well as uncertainty in the classification of unreassuring or risky FHR recordings. Given the clinical relevance of the interpretation of FHR traces as well as the role of FHR as a marker of fetal wellbeing autonomous nervous system development, many different approaches for computerized processing and analysis of FHR patterns have been proposed in the literature. The objective of this review is to describe the techniques, methodologies, and algorithms proposed in this field so far, reporting their main achievements and discussing the value they brought to the scientific and clinical community. The review explores the following two main approaches to the processing and analysis of FHR signals: traditional (or linear) methodologies, namely, time and frequency domain analysis, and less conventional (or nonlinear) techniques. In this scenario, the emerging role and the opportunities offered by Artificial Intelligence tools, representing the future direction of EFM, are also discussed with a specific focus on the use of Artificial Neural Networks, whose application to the analysis of accelerations in FHR signals is also examined in a case study conducted by the authors.
Central centrifugal cicatricial alopecia (CCCA) is the most common form of scarring alopecia among women of African ancestry. The disease is occasionally observed to affect women in families in a ...manner that suggests an autosomal dominant trait and usually manifests clinically after intense hair grooming. We sought to determine whether there exists a genetic basis of CCCA and, if so, what it is.
We used exome sequencing in a group of women with alopecia (discovery set), compared the results with those in a public repository, and applied other filtering criteria to identify candidate genes. We then performed direct sequencing to identify disease-associated DNA variations and RNA sequencing, protein modeling, immunofluorescence staining, immunoblotting, and an enzymatic assay to evaluate the consequences of potential etiologic mutations. We used a replication set that consisted of women with CCCA to confirm the data obtained with the discovery set.
In the discovery set, which included 16 patients, we identified one splice site and three heterozygous missense mutations in
in 5 patients (31%). (The approximate prevalence of the disease is up to 5.6%.)
encodes peptidyl arginine deiminase, type III (PADI3), an enzyme that post-translationally modifies other proteins that are essential to hair-shaft formation. All three CCCA-associated missense mutations in
affect highly conserved residues and are predicted to be pathogenic; protein modeling suggests that they result in protein misfolding. These mutations were found to result in reduced PADI3 expression, abnormal intracellular localization of the protein, and decreased enzymatic activity - findings that support their pathogenicity. Immunofluorescence staining showed decreased expression of PADI3 in biopsy samples of scalp skin obtained from patients with CCCA. We then directly sequenced
in an additional 42 patients (replication set) and observed genetic variants in 9 of them. A post hoc analysis of the combined data sets showed that the prevalence of
mutation was higher among patients with CCCA than in a control cohort of women of African ancestry (P = 0.002 by the chi-square test; P = 0.006 by Fisher's exact test; and after adjustment for relatedness of persons, P = 0.03 and P = 0.04, respectively).
Mutations in
, which encodes a protein that is essential to proper hair-shaft formation, were associated with CCCA. (Funded by the Ram Family Foundation and others.).
Uncombable hair syndrome (UHS), also known as “spun glass hair syndrome,” “pili trianguli et canaliculi,” or “cheveux incoiffables” is a rare anomaly of the hair shaft that occurs in children and ...improves with age. UHS is characterized by dry, frizzy, spangly, and often fair hair that is resistant to being combed flat. Until now, both simplex and familial UHS-affected case subjects with autosomal-dominant as well as -recessive inheritance have been reported. However, none of these case subjects were linked to a molecular genetic cause. Here, we report the identification of UHS-causative mutations located in the three genes PADI3 (peptidylarginine deiminase 3), TGM3 (transglutaminase 3), and TCHH (trichohyalin) in a total of 11 children. All of these individuals carry homozygous or compound heterozygous mutations in one of these three genes, indicating an autosomal-recessive inheritance pattern in the majority of UHS case subjects. The two enzymes PADI3 and TGM3, responsible for posttranslational protein modifications, and their target structural protein TCHH are all involved in hair shaft formation. Elucidation of the molecular outcomes of the disease-causing mutations by cell culture experiments and tridimensional protein models demonstrated clear differences in the structural organization and activity of mutant and wild-type proteins. Scanning electron microscopy observations revealed morphological alterations in hair coat of Padi3 knockout mice. All together, these findings elucidate the molecular genetic causes of UHS and shed light on its pathophysiology and hair physiology in general.
Throughout the world, emergency departments (ED) are characterized by overcrowding and excessive waiting times. Furthermore, the related delays significantly increase patient mortality and make ...inefficient use of resources to the detriment of the satisfaction of employees and patients. In this work, lean thinking is applied to the ED of Cardarelli Hospital of Naples with the aim of increasing patient flow, improving the processes that contribute to facilitating the flow of patients through the various stages of medical treatment and eliminating all bottlenecks (queue) as well as all activities that generate waste.
This project was performed at National Hospital A.O.R.N. A. Cardarelli of Naples. The historical times of access to the ED were analysed from January 2015 to June 2015, for a total of 16,563 records. Subsequently, starting in November 2015, corrective actions were implemented according to the Lean Approach. Data collected after the introduced improvements were collected from April 2016 to June 2016 and compared to those collected during the starting period.
The results acquired before application of the Lean Thinking strategy illustrated the as-is process with its drawbacks. An analysis of the non-added value activities was performed to identify the procedures that need to be improved. After implementation of the corrective actions, we observed a positive increase in the performance of the ED, quantified as percentages of hospitalized patients according to triage codes and waiting times.
This work demonstrates the applicability of Lean Thinking to ED processes and its effectiveness in terms of increasing the efficiency of services and reducing waste (waiting times).
In the field of electronic fetal health monitoring, computerized analysis of fetal heart rate (FHR) signals has emerged as a valid decision-support tool in the assessment of fetal wellbeing. Despite ...the availability of several approaches to analyze the variability of FHR signals (namely the FHRV), there are still shadows hindering a comprehensive understanding of how linear and nonlinear dynamics are involved in the control of the fetal heart rhythm. In this study, we propose a straightforward processing and modeling route for a deeper understanding of the relationships between the characteristics of the FHR signal. A multiparametric modeling and investigation of the factors influencing the FHR accelerations, chosen as major indicator of fetal wellbeing, is carried out by means of linear and nonlinear techniques, blockwise dimension reduction, and artificial neural networks. The obtained results show that linear features are more influential compared to nonlinear ones in the modeling of HRV in healthy fetuses. In addition, the results suggest that the investigation of nonlinear dynamics and the use of predictive tools in the field of FHRV should be undertaken carefully and limited to defined pregnancy periods and FHR mean values to provide interpretable and reliable information to clinicians and researchers.
Objective
To evaluate whether accelerated brain aging occurs in individuals with mood or psychotic disorders.
Methods
A systematic review following PRISMA guidelines was conducted. A meta‐analysis ...was then performed to assess neuroimaging‐derived brain age gap in three independent groups: (1) schizophrenia and first‐episode psychosis, (2) major depressive disorder, and (3) bipolar disorder.
Results
A total of 18 papers were included. The random‐effects model meta‐analysis showed a significantly increased neuroimaging‐derived brain age gap relative to age‐matched controls for the three major psychiatric disorders, with schizophrenia (3.08; 95%CI 2.32; 3.85; p < 0.01) presenting the largest effect, followed by bipolar disorder (1.93; 0.53; 3.34; p < 0.01) and major depressive disorder (1.12; 0.41; 1.83; p < 0.01). The brain age gap was larger in older compared to younger individuals.
Conclusion
Individuals with mood and psychotic disorders may undergo a process of accelerated brain aging reflected in patterns captured by neuroimaging data. The brain age gap tends to be more pronounced in older individuals, indicating a possible cumulative biological effect of illness burden.
Meningococcal meningitis remains a substantial cause of mortality and morbidity worldwide. Until recently, countries in the African meningitis belt were susceptible to devastating outbreaks, largely ...attributed to serogroup A Neisseria meningitidis (MenA). Vaccination with glycoconjugates of MenA capsular polysaccharide led to an almost complete elimination of MenA clinical cases. To understand the molecular basis of vaccine-induced protection, we generated a panel of oligosaccharide fragments of different lengths and tested them with polyclonal and monoclonal antibodies by inhibition enzyme-linked immunosorbent assay, surface plasmon resonance, and competitive human serum bactericidal assay, which is a surrogate for protection. The epitope was shown to optimize between three and six repeating units and to be O-acetylated. The molecular interactions between a protective monoclonal antibody and a MenA capsular polysaccharide fragment were further elucidated at the atomic level by saturation transfer difference NMR spectroscopy and X-ray crystallography. The epitope consists of a trisaccharide anchored to the antibody via the O- and N-acetyl moieties through either H-bonding or CH–π interactions. In silico docking showed that 3-O-acetylation of the upstream residue is essential for antibody binding, while O-acetate could be equally accommodated at three and four positions of the other two residues. These results shed light on the mechanism of action of current MenA vaccines and provide a foundation for the rational design of improved therapies.
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