A Novel null homozygous mutation confirms <i>CACNA2D2</i> as a gene mutated in epileptic encephalopathy

Contribution to epileptic encephalopathy (EE) of mutations in CACNA2D2, encoding α2δ-2 subunit of Voltage Dependent Calcium Channels, is unclear. To date only one CACNA2D2 mutation altering channel functionality has been identified in a single family. In the same family, a rare CELSR3 polymorphism also segregated with disease. Involvement of CACNA2D2 in EE is therefore not confirmed, while that of CELSR3 is questionable. In a patient with epilepsy, dyskinesia, cerebellar atrophy, psychomotor delay and dysmorphic features, offspring to consanguineous parents, we performed whole exome sequencing (WES) for homozygosity mapping and mutation detection. WES identified extended autozygosity on chromosome 3, containing two novel homozygous candidate mutations: c.1295delA (p.Asn432fs) in CACNA2D2 and c.G6407A (p.Gly2136Asp) in CELSR3. Gene prioritization pointed to CACNA2D2 as the most prominent candidate gene. The WES finding in CACNA2D2 resulted to be statistically significant (p = 0.032), unlike that in CELSR3. CACNA2D2 homozygous c.1295delA essentially abolished α2δ-2 expression. In summary, we identified a novel null CACNA2D2 mutation associated to a clinical phenotype strikingly similar to the Cacna2d2 null mouse model. Molecular and statistical analyses together argued in favor of a causal contribution of CACNA2D2 mutations to EE, while suggested that finding in CELSR3, although potentially damaging, is likely incidental

Detection of phylogenetically informative polymorphisms in the entire euchromatic portion of human Y chromosome from a Sardinian sample

Background: Next-Generation Sequencing methods have led to a great increase in phylogenetically useful markers within the male specific portion of the Y chromosome, but previous studies have limited themselves to the study of the X-degenerate regions. Methods: DNA was extracted from peripheral blood samples of adult males whose paternal grandfathers were born in Sardinia. The DNA samples were sequenced, genotyped and subsequently analysed for variant calling for approximately 23.1 Mbp of the Y chromosome. A phylogenetic tree was built using Network 4.6 software. Results: From low coverage whole genome sequencing of 1,194 Sardinian males, we extracted 20,155 phylogenetically informative single nucleotide polymorphisms from the whole euchromatic region, including the X-degenerate, X-transposed, and Ampliconic regions, along with variants in other unclassified chromosome intervals and in the readable sequences of the heterochromatic region. Conclusions: The non X-degenerate classes contain a significant portion of the phylogenetic variation of the whole chromosome and their inclusion in the analysis, almost doubling the number of informative polymorphisms, refining the known molecular phylogeny of the human Y chromosome

Synchronic macrophage response andPlasmodium falciparummalaria

Human Chitotriosidase (CHIT), produced by activated macrophage, is a member of the chitinase family, a group of enzymes with the capability to hydrolyze chitin. Recently plasma CHIT activity was found elevated in children with acuteP. falciparummalaria compared with healthy African children, as a consequence of macrophage activation due to the presence of parasites. In this study we recruited at the local Centre Medical Saint Camille (CMSC) of Ouagadougou, the capital of Burkina Faso, 62 African children (30 males and 32 females, aged 2–140 months; median 16.5 months), affected by acuteP. falciparummalaria, born and living in Burkina Faso. Control subjects included 140 healthy African children (79 males and 61 females) with age ranging from 10 to 100 months (median 22 months) at evaluation time. They did not show signs of acute infectious disease and their blood smears forP. falciparumwere negative. This study was approved by the local Ethical Committees of CMSC. Parents of the participating children in the study were orally informed of the scope of this research. For plasma CHIT assay, 3 ml of EDTA-blood was centrifuged and plasma samples were stored at –40°C determination by fluorimetric method at the Centre for Metabolic Diseases— University of Catania, Italy

Novel mutations in c2orf71 causing an early onset form of cone-rod dystrophy: A molecular diagnosis after 20 years of clinical follow-up

Purpose: Cone rod-dystrophies (CRDs) are pigmentary retinopathies mainly involving cones. CRDs typically present with decreased visual acuity and loss of sensitivity in the central visual field, reflecting the primary dysfunction of cones associated with night blindness and concentric visual field loss due to rod dysfunction. We describe the phenotype, natural history, and molecular analysis results of an early onset form of CRD. Methods: An otherwise healthy 25-year-old man from Sardinia, Italy, initially presented with subacute visual loss and central scotoma in both eyes. He underwent a complete ophthalmic examination, electrophysiologic testing, and genetic counseling. We first applied a candidate gene approach on ABCA4 to detect mutations; then, we performed exome sequencing (WES) on all family members to identify causative mutations. Results: The ophthalmic examination was unremarkable except the fundus examination, which revealed a well-circumscribed ring-shaped area of choroidal and RPE atrophy surrounding the fovea in the left eye and small white patches of atrophy around the fovea in the right eye. The ocular features and medical history were consistent with a diagnosis of CRD. Twenty years later, he showed a marked impairment in visual function, secondary to severe atrophic maculopathy associated with sparse pigmentary deposits. Molecular analysis identified two novel frameshift mutations in C2orf71: c.3039dupC: p.Ser1014Leufs*93 and c.1804_1805delAG:p. His603Argfs*77

Physico-Chemical Investigation and Antimicrobial Efficacy of Ozonated Oils: The Case Study of Commercial Ozonated Olive and Sunflower Seed Refined Oils

Drug resistance represents one of the great plagues of our time worldwide. This largely limits the treatment of common infections and requires the development of new antibiotics or other alternative approaches. Noteworthy, the indiscriminate use of antibiotics is mostly responsible for the selection of mutations that confer drug resistance to microbes. In this regard, recently, ozone has been raising interest for its unique biological properties when dissolved in natural oils. Ozonated oils have been reported to act in a non-specific way on microorganisms hindering the acquisition of advantageous mutations that result in resistance. Here, we focused on the antimicrobial effect of two commercial olive (OOO) and sunflower seeds (OSO) oils. Nuclear magnetic resonance spectroscopy and thermal analysis showed the change in the chemical composition of the oils after ozonation treatment. Different ozonated oil concentrations were then used to evaluate their antimicrobial profile against Candida albicans, Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Escherichia coli by agar diffusion and broth dilution methods. Cytotoxicity was also evaluated in keratinocytes and epithelial cells. Overall, our results revealed that both OOO and OSO showed a potent microbicidal effect, especially against C. albicans (IC50 = OOO: 0.3 mg/mL and OSO: 0.2 mg/mL) and E. faecalis (IC50 = OOO: 0.4 mg/mL and OSO: 2.8 mg/mL) albeit exerting a certain effect also against S. aureus and E. coli. Moreover, both OOO and OSO do not yield any relevant cytotoxic effect at the active concentrations in both cell lines. This indicates that the ozonated oils studied are not toxic for mammalian cells despite exerting a potent antimicrobial effect on specific microorganisms. Therefore, OOO and OSO may be considered to integrate standard therapies in the treatment of common infections, likely overcoming drug resistance issues

Synchronic macrophage response and <i>Plasmodium falciparum</i> malaria

Human Chitotriosidase (CHIT), produced by activated macrophage, is a member of the chitinase family, a group of enzymes with the capability to hydrolyze chitin. Recently plasma CHIT activity was found elevated in children with acute P. falciparum malaria compared with healthy African children, as a consequence of macrophage activation due to the presence of parasites. In this study we recruited at the local Centre Medical Saint Camille (CMSC) of Ouagadougou, the capital of Burkina Faso, 62 African children (30 males and 32 females, aged 2–140 months; median 16.5 months), affected by acute P. falciparum malaria, born and living in Burkina Faso. Control subjects included 140 healthy African children (79 males and 61 females) with age ranging from 10 to 100 months (median 22 months) at evaluation time. They did not show signs of acute infectious disease and their blood smears for P. falciparum were negative. This study was approved by the local Ethical Committees of CMSC. Parents of the participating children in the study were orally informed of the scope of this research. For plasma CHIT assay, 3 ml of EDTA-blood was centrifuged and plasma samples were stored at –40°C determination by fluorimetric method at the Centre for Metabolic Diseases— University of Catania, Italy

SARS-CoV-2 Evolution: Implications for Diagnosis, Treatment, Vaccine Effectiveness and Development

The COVID-19 pandemic, driven by the rapid evolution of the SARS-CoV-2 virus, presents ongoing challenges to global public health. SARS-CoV-2 is characterized by rapidly evolving mutations, especially in (but not limited to) the spike protein, complicating predictions about its evolutionary trajectory. These mutations have significantly affected transmissibility, immune evasion, and vaccine efficacy, leading to multiple pandemic waves with over half a billion cases and seven million deaths globally. Despite several strategies, from rapid vaccine development and administration to the design and availability of antivirals, including monoclonal antibodies, already having been employed, the persistent circulation of the virus and the emergence of new variants continue to result in high case numbers and fatalities. In the past four years, immense research efforts have contributed much to our understanding of the viral pathogenesis mechanism, the COVID-19 syndrome, and the host–microbe interactions, leading to the development of effective vaccines, diagnostic tools, and treatments. The focus of this review is to provide a comprehensive analysis of the functional impact of mutations on diagnosis, treatments, and vaccine effectiveness. We further discuss vaccine safety in pregnancy and the implications of hybrid immunity on long-term protection against infection, as well as the latest developments on a pan-coronavirus vaccine and nasal formulations, emphasizing the need for continued surveillance, research, and adaptive public health strategies in response to the ongoing SARS-CoV-2 evolution race

Methylenetetrahydrofolate Reductase Gene Polymorphisms in Burkina Faso: Impact on Plasma Fasting Homocysteine and after Methionine Loading Test

SUMMARY In Burkina Faso the levels of plasma homocysteine (Hcy) are lower and the methionine loading tests suggest a more effective Hcy metabolism. The polymorphisms of methylenetetrahydrofolate reductase (MTHFR) showed a relevant difference in the allele frequencies of T MTHFR-677 in young and in old subjects, while the allele frequency of C MTHFR-1298C was comparable in young and old subjects. The aim of this paper was to study the impact of the MTHFR polymorphisms on plasma fasting Hcy and after methionine loading in Burkina Faso. The young subjects with CC MTHFR-677 genotype had levels of Hcy significantly lower than CT and TT subjects. The level of Hcy in subjects who had AA, AC and CC MTHFR-1298 genotypes were comparable. The levels of Hcy after the methionine loading test were significantly higher in CT and TT MTHFR-677 genotype. These results suggest that the genetic situation in Burkina Faso is different from that of other Western countries and this guarantees the maintenance of lower plasma levels of Hcy in young and old Africans. The elevated levels of plasma Hcy in old subjects compared to young subjects, against the low prevalence of the T allele in elderly subjects is discussed. (Clin. Lab. 2007;53:XXX-XXX) KEY WORDS Burkina Faso, homocysteine, methionine loading test, MTHFR, C677T, A1298

Microsatellites and SNPs linkage analysis in a Sardinian genetic isolate confirms several essential hypertension loci previously identified in different populations

Background. A multiplicity of study designs such as gene candidate analysis, genome wide search (GWS) and, recently, whole genome association studies have been employed for the identification of the genetic components of essential hypertension (EH). Several genome-wide linkage studies of EH and blood pressure-related phenotypes demonstrate that there is no single locus with a major effect while several genomic regions likely to contain EH-susceptibility loci were validated by multiple studies. Methods. We carried out the clinical assessment of the entire adult population in a Sardinian village (Talana) and we analyzed 16 selected families with 62 hypertensive subjects out of 267 individuals. We carried out a double GWS using a set of 902 uniformly spaced microsatellites and a high-density SNPs map on the same group of families. Results. Three loci were identified by both microsatellites and SNP scans and the obtained linkage results showed a remarkable degree of similarity. These loci were identified on chromosome 2q24, 11q23.1–25 and 13q14.11–21.33. Further support to these findings is their broad description present in literature associated to EH or related phenotypes. Bioinformatic investigation of these loci shows several potential EH candidate genes, several of whom already associated to blood pressure regulation pathways. Conclusion. Our search for major susceptibility EH genetic factors evidences that EH in the genetic isolate of Talana is due to the contribution of several genes contained in loci identified and replicated by earlier findings in different human populations

A comparison of different methods to analyse data collected during time-to-exhaustion tests

Despite their widespread use in exercise physiology, time-to-exhaustion (TTE) tests present an often-overlooked challenge to researchers, which is how to computationally deal with between- and within-subject differences in exercise duration. We aimed to verify the best analysis method to overcome this problem