Modeling RTT Syndrome by iPSC-Derived Neurons from Male and Female Patients with Heterogeneously Severe Hot-Spot MECP2 Variants

Rett syndrome caused by MECP2 variants is characterized by a heterogenous clinical spectrum accounted for in 60% of cases by hot-spot variants. Focusing on the most frequent variants, we generated in vitro iPSC-neurons from the blood of RTT girls with p.Arg133Cys and p.Arg255*, associated to mild and severe phenotype, respectively, and of an RTT male harboring the close to p.Arg255*, p.Gly252Argfs*7 variant. Truncated MeCP2 proteins were revealed by Western blot and immunofluorescence analysis. We compared the mutant versus control neurons at 42 days for morphological parameters and at 120 days for electrophysiology recordings, including girls' isogenic clones. A precocious reduced morphological complexity was evident in neurons with truncating variants, while in p.Arg133Cys neurons any significant differences were observed in comparison with the isogenic wild-type clones. Reduced nuclear size and branch number show up as the most robust biomarkers. Patch clamp recordings on mature neurons allowed the assessment of cell biophysical properties, V-gated currents, and spiking pattern in the mutant and control cells. Immature spiking, altered cell capacitance, and membrane resistance of RTT neurons, were particularly pronounced in the Arg255* and Gly252Argfs*7 mutants. The overall results indicate that the specific markers of in vitro cellular phenotype mirror the clinical severity and may be amenable to drug testing for translational purposes

Case report: a typical Silver-Russell syndrome patient with hand dystonia: the valuable support of the consensus statement to the wide syndromic spectrum

The amount of Insulin Growth Factor 2 (IGF2) controls the rate of embryonal and postnatal growth. The IGF2 and adjacent H19 are the imprinted genes of the telomeric cluster in the 11p15 chromosomal region regulated by differentially methylated regions (DMRs) or imprinting centers (ICs): H19/IGF2:IG-DMR (IC1). Dysregulation due to IC1 Loss-of-Methylation (LoM) or Gain-of-Methyaltion (GoM) causes Silver–Russell syndrome (SRS) or Beckwith–Wiedemann syndrome (BWS) disorders associated with growth retardation or overgrowth, respectively. Specific features define each of the two syndromes, but isolated asymmetry is a common cardinal feature, which is considered sufficient for a diagnosis in the BWS spectrum. Here, we report the case of a girl with right body asymmetry, which suggested BWS spectrum. Later, BWS/SRS molecular analysis identified IC1_LoM revealing the discrepant diagnosis of SRS. A clinical re-evaluation identified a relative macrocephaly and previously unidentified growth rate at lower limits of normal at birth, feeding difficulties, and asymmetry. Interestingly, and never previously described in IC1_LoM SRS patients, since the age of 16, she has developed hand-writer’s cramps, depression, and bipolar disorder. Trio-WES identified a VPS16 heterozygous variant [NM_022575.4:c.2185C>G:p.Leu729Val] inherited from her healthy mother. VPS16 is involved in the endolysosomal system, and its dysregulation is linked to autosomal dominant dystonia with incomplete penetrance and variable expressivity. IGF2 involvement in the lysosomal pathway led us to speculate that the neurological phenotype of the proband might be triggered by the concurrent IGF2 deficit and VPS16 alteration

Pleural mesothelioma risk in the construction industry: a case-control study in Italy, 2000-2018

Objectives Workers in the construction industry have been exposed to asbestos in various occupations. In Italy, a National Mesothelioma Registry has been implemented more than 20 years ago. Using cases selected from this registry and exploiting existing control data sets, we estimated relative risks for pleural mesothelioma (PM) among construction workers. DesignCase-control study. SettingCases from the National Mesothelioma Registry (2000-2018), controls from three previous case-control studies. MethodsWe selected male PM incident cases diagnosed in 2000-2018. Population controls were taken from three studies performed in six Italian regions within two periods (2002-2004 and 2012-2016). Age-adjusted and period-adjusted unconditional logistic regression models were fitted to estimate odds ratios (OR) for occupations in the construction industry. We followed two approaches, one (primary) excluding and the other (secondary) including subjects employed in other non-construction blue collar occupations for >5 years. For both approaches, we performed an overall analysis including all cases and, given the incomplete temporal and geographic overlap of cases and controls, three time or/and space restricted sensitivity analyses. ResultsThe whole data set included 15 592 cases and 2210 controls. With the primary approach (4797 cases and 1085 controls), OR was 3.64 (2181 cases) for subjects ever employed in construction. We found elevated risks for blue-collar occupations (1993 cases, OR 4.52), including bricklayers (988 cases, OR 7.05), general construction workers (320 cases, OR 4.66), plumbers and pipe fitters (305 cases, OR 9.13), painters (104 cases, OR 2.17) and several others. Sensitivity analyses yielded very similar findings. Using the secondary approach, we observed similar patterns, but ORs were remarkably lower. ConclusionsWe found markedly increased PM risks for most occupations in the construction industry. These findings are relevant for compensation of subjects affected with mesothelioma in the construction industry

Infectivity in Skeletal Muscle of Cattle with Atypical Bovine Spongiform Encephalopathy

The amyloidotic form of bovine spongiform encephalopathy (BSE) termed BASE is caused by a prion strain whose biological properties differ from those of typical BSE, resulting in a clinically and pathologically distinct phenotype. Whether peripheral tissues of BASE-affected cattle contain infectivity is unknown. This is a critical issue since the BASE prion is readily transmissible to a variety of hosts including primates, suggesting that humans may be susceptible. We carried out bioassays in transgenic mice overexpressing bovine PrP (Tgbov XV) and found infectivity in a variety of skeletal muscles from cattle with natural and experimental BASE. Noteworthy, all BASE muscles used for inoculation transmitted disease, although the attack rate differed between experimental and natural cases (∼70% versus ∼10%, respectively). This difference was likely related to different prion titers, possibly due to different stages of disease in the two conditions, i.e. terminal stage in experimental BASE and pre-symptomatic stage in natural BASE. The neuropathological phenotype and PrPres type were consistent in all affected mice and matched those of Tgbov XV mice infected with brain homogenate from natural BASE. The immunohistochemical analysis of skeletal muscles from cattle with natural and experimental BASE showed the presence of abnormal prion protein deposits within muscle fibers. Conversely, Tgbov XV mice challenged with lymphoid tissue and kidney from natural and experimental BASE did not develop disease. The novel information on the neuromuscular tropism of the BASE strain, efficiently overcoming species barriers, underlines the relevance of maintaining an active surveillance

Perceptions of ecosystem services and disservices associated with open water swimming

Open water swimming is becoming an increasingly popular recreational activity. Understanding the human well-being benefits that open water swimmers derive from interaction with outdoor spaces is fundamental to environmental decision making, yet little is currently known about the benefits, threats and needs of open water swimmers in relation to the environments they swim in. In this study we provide a balanced assessment of the perceived cultural ecosystem services (CES) and ecosystem disservices (EDS) experienced by open water swimmers in the UK. Factor analysis based on an online questionnaire with 505 respondents revealed three distinct clusters of CES benefits that contribute to human well-being (‘spiritual value and interaction with nature’, ‘mental and physical health’, and ‘social bonds’). Swimming frequency, site type, gender and whether respondents were members of environmental groups affected response patterns towards CES benefits. There were five distinct clusters of EDS (‘natural hazards’, ‘security and safety (health)’, ‘security and safety (weather)’, ‘recreation’ and ‘material (biotic and abiotic)’. Key EDS ranked as an issue (by more than 50% of respondents) related to water quality, currents, rubbish and litter in water bodies and boat traffic. Swimming duration, age and swimming frequency in particular affected response patterns surrounding EDS, and to a small extent the site type. The results can be used to support the effective implementation of national and international policy, which aims to improve human well-being by encouraging use of outdoor spaces, whilst promoting the conservation and sustainable use of freshwater and marine ecosystems

Perinatal Transmission and Outcome of Neonates Born to SARS-CoV-2-Positive Mothers: The Experience of 2 Highly Endemic Italian Regions

COVID-19 is the disease caused by the novel coronavirus SARS-CoV-2, responsible of the pandemic declared in March 2020 and still ongoing. COVID-19 affects all ages but presents less complications and fatalities in children. Neonatal infections have rarely been reported worldwide, and vertical transmission is uncertain

CONFRONTO TRA ONDATE DI COVID-19 IN OPERATORI SANITARI: STUDIO RETROSPETTIVO IN UN’AZIENDA OSPEDALIERO-UNIVERSITARIA ITALIANA

L’adozione di protocolli preventivi, volti a garantire la tutela della salute degli operatori sanitari durante la pandemia COVID-19, si è dimostrata un’efficace strategia per ridurre la circolazione del virus in un setting ad elevato rischio come quello assistenziale. Durante i primi mesi della pandemia, in Italia, tali misure hanno consentito di ottenere una prevalenza estremamente bassa di infezione da SARS-CoV-2 (0.4%) tra i circa 6000 operatori sanitari dell’Azienda Ospedaliera Universitaria di Bari, uno dei principali hub COVID-19 nel Sud Italia. Tale risultato è stato, tuttavia, ottenuto durante un periodo di bassa trasmissione comunitaria del virus nella Regione Puglia

Environmental and biological monitoring of PAHs exposure in coke-oven workers at the Taranto plant compared to two groups from the general population of Apulia, Italy

Background: Polycyclic aromatic hydrocarbons (PAHs) exposure in the coke industry poses a risk for workers’ health as well as for subjects living in the plant vicinity.Objectives: To assess PAHs exposure in coke-oven workers (CW) at the Taranto plant, Apulia, and in subjects from the general population living near (NC) and far away (FC) from the plant. Methods: Exposure was assessed by personal air sampling and urinary 1-hydroxypyrene (1-OHP) measured in 100 CW, 18 NC and 15 FC. Results:Median airborne benzo[a]pyrene (BaP) levels were 152, 1.5, and 3.6 ng/m3 in CW, NC, and FC, respectively. In CW, median 1-OHP increased from 1.45 to 1.96 μg/g creatinine (crt) during the work shift (p>0.05); in NC and FC, 1-OHP levels were 0.56 and 0.53 μg/g crt. No significant differences between NC and FC for both air and urinary indices were found. BaP exposure in CW exceeded the recently proposed German acceptable (70 ng/m3) and tolerable (700 ng/m3) risk-based limit values in 82 and 11% of subjects, respectively. In NC and FC, BaP exposure exceeded the European target value for ambient air (1 ng/m3) in 67 and 60% of subjects, respectively. Biomonitoring showed that 21% of CW had 1-OHP levels higher than the proposed biological limit value for the coke-oven industry (4.4 μg/g crt), while 93% of FC, and 88% of NC, had 1-OHP levels exceeding the Italian reference value (0.3 μg/g crt). Among non-smokers, a linear regression between 1-OHP and BaP (Pearson value r=0.65, p<0.05) allowed us to estimate levels of 1.2 and 1.9 μg/g crt for 1-OHP end-of-shift corresponding to acceptable and tolerable limit values. Conclusions: Although lower than in the past, PAHs exposure in the coke plant still poses a health risk for workers and the general population and requires further efforts to improve workplace conditions

Lost in the dark: Antipatharia-Symbiodiniaceae association in the deep waters of the Red Sea

Black corals (Hexacorallia: Antipatharia) are a major component of mesophotic and deep marine ecosystems. Due to their preference for light deprived environments, black corals have historically been considered azooxanthellate, yet recent works have found them in association with dinoflagellates of the family Symbiodiniaceae down to 396 m depth. While corals and Symbiodiniaceae generally establish a symbiotic relationship in shallow water environments, the implications of this association is less well understood at deeper depths, where low light penetration cannot sustain efficient photosynthetic activity for the algae. However, Symbiodinaceae are not obligate autotrophs, and their capacity for heterotrophic feeding categorizes them as mixotrophs. In this study, we investigated the presence and diversity of Symbiodiniaceae associated with the deep-sea black coral Bathypathes thermophila (Antipatharia: Schizopathidae), collected from 204 to 655 m depth in the Saudi Arabian Red Sea. Using high-throughput sequencing of the ITS2 region, we report (1) the deepest record to date of Symbiodiniaceae associated with an anthozoan from 655 m, and (2) the first Red Sea record of Antipatharia in association with Symbiodiniaceae. Our analyses revealed that 14 out of 27 colonies of B. thermophila were associated with Symbiodiniaceae of the genera Cladocopium and Durusdinium. We unveiled 16 novel ITS2 type profiles, possibly unique to black corals and/or to these depths, along with seven profiles that were already known from shallow-water hard corals. No significant pattern was detected in terms of community diversity in relation to depth or sampling locality. Our study supports the existence of black corals-Symbiodiniaceae association and warrants further research to better understand the evolutionary processes and physiological mechanisms driving this association, specifically in light deprived environments

From the shallow to the mesophotic: a characterization of Symbiodiniaceae diversity in the Red Sea NEOM region

IntroductionThe northern Red Sea has been coined a refuge for reef corals due to the exceptional thermal tolerance of these organisms. With ocean warming threatening coral reefs worldwide, a panoptic characterization of corals living in extreme conditions may provide insights into future responses of corals to environmental change. Among other factors, the genotype of the endosymbiotic algae in the family Symbiodiniaceae has been shown to have major implications on the distribution and resilience of their coral hosts. In this study, we aim at genotyping the Symbiodiniaceae communities associated with three depth generalist and one depth specialist coral species, characterized by the ability to withstand environmental conditions that are apparently limiting for other corals and occurring in a unique geographical region.MethodsWe sampled 50 corals from the northern Saudi Arabian Red Sea and the Gulf of Aqaba, covering a 97 m bathymetric gradient. We used high-throughput ITS2 gene sequencing and recovered different patterns of host–algal associations.Results and discussionThe majority of the recovered algal genotypes appeared host- and environment-specific, while others were more widely distributed. At large, coral specimens were overwhelmingly associated with symbionts from the genus Cladocopium and specifically with many previously undescribed genotypes. This suggests the selection of specific genotypes, which might confer resistance and/or resilience to their host counterparts. Interestingly, we found a limited association with Durusdinium spp. and other known tolerant taxa in mesophotic corals in the northern Red Sea, but not in the Gulf of Aqaba. The broad absence of Durusdinium spp., typically ascribed to be stress tolerant, warrants further investigation into Symbiodiniaceae species that convey environmental resilience. Our data will serve as a baseline to explore the occurrence of specific symbionts that might be contributing to coral acclimation and adaptation and to assay how biodiversity might be impacted if subjected to increasing stressors