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Depression is characterized by a loss of synaptic connections in regions of the brain involved in emotional regulation and cognitive function, and antidepressant treatments generally result in a compensatory increase in the number of these connections. Previous studies have shown that a single sub-anesthetic dose of ketamine, a rapid-acting antidepressant, increases synaptic connectivity in excitatory neurons projecting from the prefrontal cortex to other brain regions involved in cognitive regulation and function, countering the destabilization and loss of synapses observed in depression. However, there is little understanding of the molecular pathways and mechanisms involved in synapse restoration at the synapse structure level. Due to the role of proteins involved in synapse formation and maintenance, we sought to determine whether ketamine’s antidepressant mechanism is dependent on these proteins in projecting neurons of excitatory circuits that contribute to emotional regulation. Mice were divided into groups with or without chronic stress and with conditional knockouts for the proteins of interest. They were then subjected to a series of behavioral tests and their brain tissue was analyzed for changes in excitatory synapse number. We found that ketamine’s promotion of synapse formation and sustained antidepressant-like effect on behavior was hindered in knockout mice. These findings revealed that ketamine is dependent on proteins involved in synapse formation and maintenance for its rescue of depressive-like behavior and restoration of excitatory synaptic connectivity.

The authors thank all the people involved in collecting/providing material or organising sampling campaigns: Peter Schuchert (MHNG, Switzerland), Tullia Isotta Terraneo (KAUST, Saudi Arabia), Malek Amr Gusti (KAUST, Saudi Arabia), Timothy Ravasi (OIST, Japan), the captain and crew of the MV Dream-Master (Saudi Arabia), the KAUST Coastal and Marine Resources Core Lab, Inga Dehnert (UNIMIB, Italy), Nicholas WL Yap (NUS, Singapore), Sudhanshi S Jain (NUS, Singapore), Stephen Keable (Australian Museum), Penny Berents (Australian Museum), Anne Hoggett (Australian Museum), Lyle Vail (Australian Museum). Additionally, we wish to thank Leen P. van Ofwegen (Naturalis, The Netherlands) for his valuable help in identifying the octocoral Paralemnalia sp., Peter Schuchert for his comments on an earlier version of the manuscript, and two anonymous referees for their thorough revision of this work. Permissions relevant to undertake the research have been obtained from the applicable governmental agencies. Fieldwork at St. Eustatius was funded through a Martin Fellowship from Naturalis Biodiversity Center to SM, while logistic support was supplied by St. Eustatius Marine Parks (STENAPA), the Caribbean Netherlands Science Institute (CNSI) and Scubaqua Dive Centre. Samples from Eilat (Israel) were collected during the HyDRa Project funded by the EU FP7 Research Infrastructure Initiative ‘ASSEMBLE’ (Grant #227799) to DP. Financial support to DP for collecting samples at Lizard Island (Australia) was provided by the 2018 John and Laurine Proud Fellowship and the Australian Museum’s Lizard Island Research Station. Fieldwork in Mozambique was conducted during the Green Bubbles financed by EU’s H2020 research and innovation programme to DP, under the Marie Sklodowska-Curie grant agreement no 643712 (Permit n° 09/2018 ANAC). Fieldwork in Singapore was partially funded by the National Research Foundation, Prime Minister’s Office, Singapore under its Marine Science R&D Programme (MSRDP-P03) to DH.

Microplastics pollution differentially impacts coral reef systems, by threatening corals physically, through physiological distress and by increasing diseases. However, most of the studies to date have focused on scleractinian corals. The present work reports for the first time the patterns of microplastic ingestion and adhesion by the alcyonacean Coelogorgia palmosa. Feeding and adhesion tests were carried out with various concentrations of polyethylene microbeads. Results showed a wide range of surface adhesion, ranging from 3 to 1573 microbeads per coral fragment, suggesting that adhesion driven by mucus is the main mechanism of microplastic trapping. Polyethylene was ingested by 60% of coral fragments, and the average number of ingested microbeads was much lower compared to scleractinian corals. Considering the ecological importance of soft corals in coral reef ecosystems, specific attention regarding microplastic pollution effects on this taxon is recommended.

An integrated approach using morphological and genetic data is needed to disentangle taxonomic uncertainties affecting the hydrozoan families Sphaerocorynidae and Zancleopsidae. Here we used this approach to accurately characterise species in these families, identify the previously unknown polyp stages of the genera Euphysilla and Zancleopsis, which were originally described exclusively based on the medusa stages, describe a new sphaerocorynid genus and species, and assess the phylogenetic position of the two families within the Capitata. The monotypic genus Astrocoryne was found to be a synonym of Zancleopsis. Astrocoryne cabela was therefore transferred to the genus Zancleopsis as Zancleopsis cabela comb. nov. The new polyp-based genus and species Kudacoryne diaphana gen. nov. sp. nov. was erected within the Sphaerocorynidae. Both taxa are primarily based on genetic data, but the introduction of this new genus was made necessary by the fact that it clustered with the genera Heterocoryne and Euphysilla, despite showing Sphaerocoryne-like polyps. Interestingly, the species analysed in this work showed contrasting biogeographical patterns. Based on our data and literature records, some species appear to have a wide circumtropical range, whereas others are limited to few localities. Overall, these results lay the ground for future investigations aimed at resolving the taxonomy and systematics of these two enigmatic families.

A growing number of emerging SARS-CoV-2 variants is being identified worldwide, potentially impacting the effectiveness of current vaccines. We report the data obtained in several Italian regions involved in the SARS-CoV-2 variant monitoring from the beginning of the epidemic and spanning the period from October 2020 to March 2021.

Background: If analytical antiretroviral-treatment (ART) interruption (ATI) might significantly impact quantitative or qualitative peripheral-total HIV-DNA is still debated. Methods: Six chronically HIV-1 infected patients enrolled in APACHE-study were analysed for peripheral-total HIV-DNA and residual viremia, major-resistance-mutations (MRMs) and C2-V3-C3 evolution at pre-ATI (T1), during ATI (T2) and at achievement of virological success after ART-resumption (post-ATI, T3). These data were obtained at three comparable time-points in five chronically HIV-1 infected patients on suppressive ART for ≥1 year, enrolled in MODAt-study. Results: At T1, APACHE and MODAt individuals had similar peripheral-total HIV-DNA and residual viremia (p = 0.792 and 0.662, respectively), and no significant changes for these parameters were observed between T1 and T3 in both groups. At T1, 4/6 APACHE and 2/5 MODAt carried HIV-DNA MRMs. MRMs disappeared at T3 in 3/4 APACHE. All disappearing MRMs were characterized by T1 intra-patient prevalence &lt, 80%, and mainly occurred in APOBEC3-related sites. All MRMs persisted over-time in the 2 MODAt. C2-V3-C3 genetic-distance significantly changed from T1 to T3 in APACHE individuals (+0.36[0.11–0.41], p = 0.04), while no significant changes were found in MODAt. Accordingly, maximum likelihood trees (bootstrap &gt, 70%) and genealogical sorting indices (GSI &gt, 0.50 with p-value &lt, 0.05) showed that T1 C2-V3-C3 DNA sequences were distinct from T2 and T3 viruses in 4/6 APACHE. Virus populations at all three time-points were highly interspersed in MODAt. Conclusions: This pilot study indicates that short ATI does not alter peripheral-total HIV-DNA burden and residual viremia, but in some cases could cause a genetic diversification of peripheral viral reservoir in term of both MRMs rearrangement and viral evolution.

Scleractinian corals provide habitats for a broad variety of cryptofauna, which in turn may contribute to the overall functioning of coral symbiomes. Among these invertebrates, hydrozoans belonging to the genus Zanclea represent an increasingly known and ecologically important group of coral symbionts. In this study, we analysed 321 Zanclea colonies associated with 31 coral genera collected from 11 localities across the Indo-Pacific and Caribbean regions, and used a multi-disciplinary approach to shed light on the evolution and biogeography of the group. Overall, we found high genetic diversity of hydrozoans that spans nine clades corresponding to cryptic or pseudo-cryptic species. All but two clades are associated with one or two coral genera belonging to the Complex clade, whereas the remaining ones are generalists associated with both Complex and Robust corals. Despite the observed specificity patterns, no congruence between Zanclea and coral phylogenies was observed, suggesting a lack of coevolutionary events. Most Zanclea clades have a wide distribution across the Indo-Pacific, including a generalist group extending also into the Caribbean, while two host-specific clades are possibly found exclusively in the Red Sea, confirming the importance of this peripheral region as an endemicity hotspot. Ancestral state reconstruction suggests that the most recent common ancestor of all extant coral-associated Zanclea was a specialist species with a perisarc, occurring in what is now known as the Indo-Pacific. Ultimately, a mixture of geography- and host-related diversification processes is likely responsible for the observed enigmatic phylogenetic structure of coral-associated Zanclea.

Twenty years of research have led to the model, acknowledged by the 2013 Nobel Prize in Physiology and Medicine, that soluble N -ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) mediate the fusion of intracellular membranes. SNAREs are also able to recruit other molecules playing essential roles in the spatial and temporal regulation of vesicular traffic, and they now even appear as multitask proteins in intracellular vesicle biogenesis, transport and fusion. Here we focus on recent findings at the molecular level which suggest how SNAREs operate as membrane fusion nanomachines that are finely tuned to allow fusion to occur at the right time and place, and how they are integrated into complex molecular networks of interaction.

The natural ends of linear chromosomes resemble those of accidental double-strand breaks (DSBs). DSBs induce a multifaceted cellular response that promotes the repair of lesions and slows down cell cycle progression. This response is not elicited at chromosome ends, which are organized in nucleoprotein structures called telomeres. Besides counteracting DSB response through specialized telomere-binding proteins, telomeres also prevent chromosome shortening. Despite of the different fate of telomeres and DSBs, many proteins involved in the DSB response also localize at telomeres and participate in telomere homeostasis. In particular, the DSB master regulators Tel1/ATM and Mec1/ATR contribute to telomere length maintenance and arrest cell cycle progression when chromosome ends shorten, thus promoting a tumor-suppressive process known as replicative senescence. During senescence, the actions of both these apical kinases and telomere-binding proteins allow checkpoint activation while bulk DNA repair activities at telomeres are still inhibited. Checkpoint-mediated cell cycle arrest also prevents further telomere erosion and deprotection that would favor chromosome rearrangements, which are known to increase cancer-associated genome instability. This review summarizes recent insights into functions and regulation of Tel1/ATM and Mec1/ATR at telomeres both in the presence and in the absence of telomerase, focusing mainly on discoveries in budding yeast.

Toxoplasmosis acquired in early pregnancy is a potentially severe complication for the fetus. Evaluating the risk of transplacental infection in pregnant women accessing the Tuscany Reference Center for Infectious Diseases in Pregnancy during the last 20 years with suspected or confirmed toxoplasmosis acquired in early pregnancy was the aim of the study.We retrospectively enrolled all pregnant women undergoing amniocentesis for toxoplasmosis acquired in the first 16 gestational weeks in the period 1999-2019, comparing patients with certain acute infection (seroconversion occurred in pregnancy, CAIP) with those with suspected acute infection (IgG positive with low/intermediate IgG avidity index, SAIP).237 patients were enrolled, 187 (78.9%) with SAIP and 50 (21.1%) with CAIP. Specific IgM was detected in 47.5% and 76.7% (p-value 0.001), and the mean IgG avidity index was 22.7% and 7.1% (p-value 0.001) in the SAIP and in the CAIP group, respectively. The mean delay from diagnosis to antibiotic initiation was 14.6 in SAIP and 11 days in CAIP group. Toxoplasma DNA was detected in the amniotic fluid in one case in a patient with CAIP. Excluding 24 newborns with not available data, prevalence of congenital infection was 0.47% [1/213 (95% CI 0.08%-2.61%)], 0% [0/178 (95% CI 0%-2.11%)] in SAIP and 2.8% [1/35 (95% CI 0.51%-14.53%)] in CAIP group.Toxoplasmosis acquired in early pregnancy has a low risk of fetal infection. Actively discussing case-by-case amniocentesis indication with patients, especially when a recent toxoplasmosis is not properly confirmed, is desirable.

Human activities may change beta diversity—the spatial variation in species composition—in different ways. Positive and negative trends in beta diversity are referred as biotic differentiation and homogenization, respectively. In this context, river damming is likely to be a major cause of changes in beta diversity over time. Here, we evaluated the impact of damming on zooplankton beta diversity in two Brazilian reservoirs. We predicted that damming would cause biotic differentiation due to the creation of areas with different hydrological conditions, which would allow the colonization and population growth of species belonging to different zooplankton groups. Our results for the total zooplankton community were consistent with the hypothesis of biotic differentiation, either due to the increased mean beta diversity or due to the tendency of increasing beta diversity over time after damming. An indicator species analysis also showed that a large proportion of taxa that can be categorized as euplanktonic were mainly indicators of the period after damming, whereas the opposite was true for testate amoebae. Increased beta diversity should be interpreted as an impact of damming. However, we speculate that, under a process of water quality deterioration, biotic homogenization is likely to occur, reversing the patterns we observed.

Bark and wood-boring beetles feeding on coniferous and deciduous trees in different forest ecosystems are often associated with various species of fungi. In 2019, widespread attacks of Liparthrum colchicum Semenov (Coleoptera Curculionidae Scolytinae) were observed on Laurus nobilis L. in Tuscany (Italy). Samples of colonized terminal twigs were collected to investigate the presence of phytopathogenic fungi associated with the scolytid. Two different colonies of the Geosmithia genus were identified as Geosmithia pallida and Geosmithia langdonii. To our knowledge this is the first report of G.pallida and G. langdonii associated with L. colchicum on Bay tree, in Italy.

The human dopamine transporter gene SLC6A3 has been consistently implicated in several neuropsychiatric diseases but the disease mechanism remains elusive. In this risk synthesis, we have concluded that SLC6A3 represents an increasingly recognized risk with a growing number of familial mutants associated with neuropsychiatric and neurological disorders. At least five loci were related to common and severe diseases including alcohol use disorder (high activity variant), attention-deficit/hyperactivity disorder (low activity variant), autism (familial proteins with mutated networking) and movement disorders (both regulatory variants and familial mutations). Association signals depended on genetic markers used as well as ethnicity examined. Strong haplotype selection and gene-wide epistases support multimarker assessment of functional variations and phenotype associations. Inclusion of its promoter region’s functional markers such as DNPi (rs67175440) and 5’VNTR (rs70957367) may help delineate condensate-based risk action, testing a locus-pathway-phenotype hypothesis for one gene-multidisease etiology.

Canis Major OB1 (CMa OB1) is a Galactic stellar association with a very intriguing star-formation scenario. There are more than two dozen known star clusters in its line of sight, but it is not clear which ones are physically associated with CMa OB1. We use a clustering code that employs 5-dimensional data from the Gaia DR2 catalogue to identify physical groups and obtain their astrometric parameters and, in addition, we use two different isochrone-fitting methods to estimate the ages of these groups. We find 15 stellar groups with distances between 570 pc and 1650 pc, including 10 previously known and 5 new open cluster candidates. Four groups, precisely the youngest ones ($, Comment: 23 pages, 28 figures, 7 tables

This study contains predictions for mass spectrometry and plasma instrument measurements during upcoming Venus flybys of BepiColombo and Solar Orbiter and discusses the possibility of a phosphine detection with mass spectrometry in Venus’ upper atmosphere. The results extend the ones published previously in Gruchola et al. (2019) , where predictions for the proposed ESA mission EnVision and the Venus flyby of JUICE were included. Both the Venus flyby of BepiColombo and the one of Solar Orbiter will take place in August 2021, only 1 day apart. BepiColombo, carrying the neutral mass gas spectrometer STROFIO, could probe the atmosphere around closest approach and obtain data on the thermal and hot neutral particle populations in the upper atmosphere. According to this study, the thermal hydrogen population as well as the hot species H, C, N and O, including isotopes, should be visible to STROFIO. Especially data on the abundance of the hot species would yield important insight into the planetary escape processes. The Solar Orbiter on the other hand carries the plasma instrument SWA-HIS, designed to measure the energetic solar wind ions. During its second Venus gravity assist it will traverse the magnetosheath, the region between bow shock and ion composition boundary, where in addition to the solar wind ions energetic planetary ions are present. The planetary pickup ions can be measured by SWA-HIS, providing important information on the outflow of planetary ions and the ionization processes in the upper atmosphere itself. The recent reports of phosphine in Venus’ cloud decks with an abundance of 20 ppb at 80 km probably overestimate the actual PH3 abundance, as the data are currently being reanalyzed (Greaves et al., 2021). However, even with this upper limit of 20 ppb a phosphine detection with a mass spectrometer e.g. on-board ESA’s proposed EnVision mission, seems unlikely. To resolve the PH3 peak and the 16O18O peak a mass resolution of almost 10’000 is required, and the PH2D peak is masked by the Cl fragment peak of HCl. Furthermore, NGMS on-board Pioneer Venus with a mass resolution of around 440 did most likely not detect phosphine, as it scanned only a few masses in the mass range of interest where more abundant species than phosphine are present.

In order to develop a more sensitive and reliable method for detection of serum antibodies against Mycoplasma hyopneumoniae infection in pigs, six recombinant proteins of M. hyopneumoniae (P102, P95, P46, P97 like, Lppt, and hypothetical P987) were used for the standardization of an indirect enzyme-linked immunosorbent assay (ELISA). The proteins were evaluated against 50 sera of the specific pathogen-free and 50 sera of pigs with lesions suggestive of infection. The sensitivity was 88%, 86%, 78%, 74%, 66%, and 60% for the proteins P102, P95, P46, P97 like, Lppt, and hypothetical protein P987, respectively. Moreover, the proteins were used to establish the seroprevalence in two different commercial herds (254 sera pigs from farm considered free of M. hyopneumoniae and 246 from farm with clinical signs of enzootic pneumonia and positive serology for M. hyopneumoniae) and the positive rate was 65.2% for P95, 54.6% for P102, 40.2% for P46, 37.2% for P97 like, 17.4% for the hypothetical P987, and 14% for Lppt protein. In addition, the ELISA with six recombinant proteins was compared to commercial HerdCheck kit using 118 random pig sera samples and the results showed that ELISA with recombinant proteins were more sensitive than the commercial test. These data show that the recombinant proteins P95 and P102 are potential targets to be used in diagnostic tests to detect antibodies against M. hyopneumoniae. Although more studies are necessary, this study provides insights that these recombinant proteins can be useful in epidemiological investigations and as potential biomarkers in differentiating infected animals from those vaccinated.

Background Environmental fluctuation during embryonic and fetal development can permanently alter an organism’s morphology, physiology, and behaviour. This phenomenon, known as developmental plasticity, is particularly relevant to reptiles that develop in subterranean nests with variable oxygen tensions. Previous work has shown hypoxia permanently alters the cardiovascular system of snapping turtles and may improve cardiac anoxia tolerance later in life. The mechanisms driving this process are unknown but may involve epigenetic regulation of gene expression via DNA methylation. To test this hypothesis, we assessed in situ cardiac performance during 2 h of acute anoxia in juvenile turtles previously exposed to normoxia (21% oxygen) or hypoxia (10% oxygen) during embryogenesis. Next, we analysed DNA methylation and gene expression patterns in turtles from the same cohorts using whole genome bisulfite sequencing, which represents the first high-resolution investigation of DNA methylation patterns in any reptilian species. Results Genome-wide correlations between CpG and CpG island methylation and gene expression patterns in the snapping turtle were consistent with patterns observed in mammals. As hypothesized, developmental hypoxia increased juvenile turtle cardiac anoxia tolerance and programmed DNA methylation and gene expression patterns. Programmed differences in expression of genes such as SCN5A may account for differences in heart rate, while genes such as TNNT2 and TPM3 may underlie differences in calcium sensitivity and contractility of cardiomyocytes and cardiac inotropy. Finally, we identified putative transcription factor-binding sites in promoters and in differentially methylated CpG islands that suggest a model linking programming of DNA methylation during embryogenesis to differential gene expression and cardiovascular physiology later in life. Binding sites for hypoxia inducible factors (HIF1A, ARNT, and EPAS1) and key transcription factors activated by MAPK and BMP signaling (RREB1 and SMAD4) are implicated. Conclusions Our data strongly suggests that DNA methylation plays a conserved role in the regulation of gene expression in reptiles. We also show that embryonic hypoxia programs DNA methylation and gene expression patterns and that these changes are associated with enhanced cardiac anoxia tolerance later in life. Programming of cardiac anoxia tolerance has major ecological implications for snapping turtles, because these animals regularly exploit anoxic environments throughout their lifespan.

Approximately one‐third of randomly produced knockout mouse lines produce homozygous offspring, which fail to survive the perinatal period. The majority of these die around or after embryonic day (E)14.5, presumably from cardiovascular insufficiency. For diagnosing structural abnormalities underlying death and diseases and for researching gene function, the phenotype of these individuals has to be analysed. This makes the creation of reference data, which define normal anatomy and normal variations the highest priority. While such data do exist for the heart and arteries, they are still missing for the venous system. Here we provide high‐quality descriptive and metric information on the normal anatomy of the venous system of E14.5 embryos. Using high‐resolution digital volume data and 3D models from 206 genetically normal embryos, bred on the C57BL/6N background, we present precise descriptive and metric information of the venous system as it presents itself in each of the six developmental stages of E14.5. The resulting data shed new light on the maturation and remodelling of the venous system at transition of embryo to foetal life and provide a reference that can be used for detecting venous abnormalities in mutants. To explore this capacity, we analysed the venous phenotype of embryos from 7 knockout lines (Atp11a, Morc2a, 1700067K01Rik, B9d2, Oaz1, Celf4 and Coro1c). Careful comparisons enabled the diagnosis of not only simple malformations, such as dual inferior vena cava, but also complex and subtle abnormalities, which would have escaped diagnosis in the absence of detailed, stage‐specific referenced data., We provide high‐quality descriptive and metric information on the normal anatomy of the venous system of E14.5 mouse embryos. Our data shed new light on the maturation and remodelling of the venous system at the transition of embryo to foetal life and provide a reference that can be used for detecting venous abnormalities in mutants. Using our data, we were able to diagnose not only simple malformations but also complex and subtle abnormalities of embryos of seven knockout lines.

Context To optimise litter quality, the use of gypsum (CaSO4·2H2O) may be a good strategy. Brazil is the largest exporter of chicken meat in the world. The Asian market, in particular, demands chicken feet; however, quality standards for export demand that the feet have to be free (or almost free) of injuries. It is therefore essential to reduce the incidence of footpad lesions. Aims To determine whether the replacement of lime (Ca(OH)2) by gypsum in poultry broiler litter improves litter quality, pododermatitis scores, animal performance and health. Methods In total, 320, 1-day-old male Cobb 500 broilers were divided into the following four groups: control group in which no gypsum or lime were added to the litter (TC); group with 0.5 kg of lime/m2 of litter (TL 0.5); group with 0.5 kg of gypsum/m2 of litter (TG 0.5); and group with 1.0 kg of gypsum/m2 of litter (TG 1.0). The pH, dry matter, water activity and surface temperature of the litter were measured on Days 21, 35 and 42. Animal performance parameters were measured on Days 1, 21, 35 and 42; blood collection was performed on Day 42 for biochemical and haematological analysis, in addition to excreta samples and litter for bacterial counts. At the end of the experiment, 32 broilers were humanely killed for tissue analysis (liver and intestine) and histopathology. The degrees of footpad injury (Day 42) were graded using a 5-point scale, from 0 (no injury), to 1, 2, 3 and 4 (severe injury). Key results No differences were found between treatments regarding performance (P > 0.05). With respect to intestinal parameters, there were significant differences in villus height and crypt depth, with TL 0.5, and TG 1.0 showing larger villi than did the control group. The shallowest crypt depths were seen in TC and TG 0.5. Litter dry matter content was significantly higher than in the control (TC) at Day 35 in TG 0.5, and at Day 42 in the TG 1.0 treatments. The litter temperature was significantly higher in the TL 0.5 treatment than in the TG 1.0 at 21 days, but they were not different from that in the control. At Day 35, both TG treatments had significantly lower litter temperature than did the control group, but no differences were observed at Day 42. The treatments had no effect on haematology or biochemical properties, nor on litter or bacterial counts in excreta. Broilers raised on litter treated with gypsum showed 50% fewer Grade 3 and 4 footpad lesions than did the control animals on Day 42, an important finding mainly for lesion grades between 3 and 4 that could prevent export of feet. Conclusions Gypsum at 0.5 kg/m2 showed the best results regarding litter quality, reducing the incidence of footpad lesions without compromising animal health or performance. Implications Applying gypsum to litter at 0.5 kg/m2 reduces the number of broilers with pododermatitis, an economic benefit to the poultry industry.

Interspecific associations are common in coral reefs, but those involving hydrozoans and octocorals have not been widely investigated. The hydroid Pteroclava krempfi (Hydrozoa, Cladocorynidae) lives in association with different soft coral taxa (Alcyonacea), showing a widespread distribution. However, very little information is available on the ecology of these relationships. Here, we tested for differences in the taxon-specific prevalence and habitat preference of the symbiosis and determined ecological traits of the P. krempfi-host associations in central Red Sea reefs. P. krempfi was found associated with the alcyonacean genera Lobophytum, Rhytisma, Sarcophyton and Sinularia, updating its host range and geographic distribution. The symbiosis prevalence was high in the area and especially at inshore sites compared to midshore and offshore sites. Rhytisma was the most common host, while the association with Lobophytum showed the lowest taxon-specific prevalence. P. krempfi did not show a clear preference for a specific alcyonacean size, and an increase in host size automatically led to an increase in the surface occupied by hydrozoans, although they rarely colonized more than 50% of the upper surface of the host. The spatial distribution of the hydroids on the host surface appeared related to the host genus and size as well as to the coverage of the hydroids. Despite the nature of this symbiosis requiring further investigation, P. krempfi did not seem to play a role in affecting the bleaching susceptibilities of the host colonies. The study shows that the Red Sea coral reef symbioses are more widespread than previously known and therefore deserve more attention.

Corals show spatial acclimatisation to local environment conditions. However, the various cellular mechanisms involved in local acclimatisation and variable bleaching patterns in corals remain to be thoroughly understood. In this study, the modulation of a protein implicated in cellular heat stress tolerance, the heat shock protein 70, was compared at both gene (hsp70) and protein (Hsp70) expression level in bleaching tolerant near-coast Acropora muricata colonies and bleaching susceptible reef colonies, in the lagoon of Belle Mare (Mauritius). The relative Hsp70 levels varied significantly between colonies from the two different locations, colonies having different health conditions and the year of collection. Before the bleaching event of 2016, near-coast colonies had higher basal levels of both Hsp70 gene and protein compared to reef colonies. During the bleaching event, the near-coast colonies did not bleach and had significantly higher relative levels of both Hsp70 gene and protein compared to bleached reef colonies. No significant genetic differentiation between the two studied coral populations was observed and all the colonies analysed were associated with Symbiodiniaceae of the genus Symbiodinium (Clade A) irrespective of location and sampling period. These findings provide further evidence of the involvement of Hsp70 in conferring bleaching tolerance to corals. Moreover, the consistent expression differences of Hsp70 gene and protein between the near-coast and reef coral populations in a natural setting indicate that the modulation of this Hsp is involved in local acclimatisation of corals to their environments.

Metabarcoding studies using environmental DNA (eDNA) and high‐throughput sequencing (HTS) are rapidly becoming an important tool for assessing and monitoring marine biodiversity, detecting invasive species, and supporting basic ecological research. Several barcode loci targeting teleost fish and elasmobranchs have previously been developed, but to date primer sets focusing on other marine megafauna, such as marine mammals, have received less attention. Similarly, there have been few attempts to identify potentially “universal” barcode loci which may be informative across multiple marine vertebrate orders. Here we describe the design and validation of two new sets of primers targeting hypervariable regions of the vertebrate mitochondrial 12S and 16S rRNA genes, which have conserved priming sites across virtually all cetaceans, pinnipeds, elasmobranchs, boney fish, sea turtles, and birds, and amplify fragments with consistently high levels of taxonomically diagnostic sequence variation. “In silico” validation using the OBITOOLS software showed our new barcode loci outperformed most existing vertebrate barcode loci for taxon detection and resolution. We also evaluated sequence diversity and taxonomic resolution of the new barcode loci in 680 complete marine mammal mitochondrial genomes demonstrating that they are effective at resolving amplicons for most taxa to the species level. Finally, we evaluated the performance of the primer sets with eDNA samples from aquarium communities with known species composition. These new primers will potentially allow surveys of complete marine vertebrate communities in single HTS metabarcoding assessments, simplifying workflows, reducing costs, and increasing accessibility to a wider range of investigators.

Coral diseases are emerging as a major threat to coral reefs worldwide, and although many of them have been described, knowledge on their epizootiology is still limited. This is the case of the Halofolliculina ciliate infections, recognized as the skeletal eroding band (SEB) and Caribbean ciliate infection (CCI), two diseases caused by ciliates belonging to the genus Halofolliculina (Class Heterotrichea). Despite their similar macroscopic appearance, the two diseases are considered different and their pathogens have been hypothesized to belong to different Halofolliculina species. In this work, we analysed the morphology and genetic diversity of Halofolliculina ciliates collected in the Caribbean Sea, Red Sea and Indo-Pacific Ocean. Our analyses showed a strong macroscopic similarity of the lesions and similar settlement patterns of the halofolliculinids from the collection localities. In particular, the unique erosion patterns typical of the SEB were observed also in the Caribbean corals. Fine-scale morphological and morphometric examinations revealed a common phenotype in all analysed ciliates, unequivocally identified as Halofolliculina corallasia. Phylogenetic analyses based on nuclear and mitochondrial (COI) molecular markers consistently found all samples as monophyletic. However, although the nuclear marker displayed an extremely low intra-specific diversity, consistent with the morphological recognition of a single species, the analyses based on COI showed a certain level of divergence between samples from different localities. Genetic distances between localities fall within the intra-specific range found in other heterotrich ciliates, but they may also suggest the presence of a H. corallasia species complex. In conclusion, the presented morpho-molecular characterization of Halofolliculina reveals strong similarities between the pathogens causing SEB and CCI and call for further detailed studies about the distinction of these two coral diseases.

Food-item authentication and traceability is an issue of primary concern, due to both socio-economical and health implications. DNA-based methods are increasingly being recognised as powerful tools to assess the reliability of supplier labels for any type of food. This is especially true for products characterised by a short shelf life and high-processing supply chain, such as seafood. In this work, a DNA barcoding approach was applied to assess the accuracy of species labelling in 150 cephalopod seafood products sold in the Italian market. Overall, high levels of mislabelling in squid, cuttlefish, and octopus items were identified, and in some cases, even species not included in the current food Regulations. Additionally, an application of the recently developed naked-eye detection tool ‘NanoTracer’, consisting in the combination of DNA barcoding with gold nanoparticle-based, was demonstrated to authenticate common cuttlefish (Sepia officinalis) seafood. The primer pairs used to set the fast detection system for S. officinalis were designed based on the most comprehensive DNA barcoding (COI and 16s rRNA) datasets ever assembled for cephalopods, assuring the specificity of the method. ‘NanoTracer’ allowed a simple, rapid, accurate and cost-effective authentication, revealing its potential adaptability to any type of seafood and other food categories.

Funder: British Heart Foundation; Id: http://dx.doi.org/10.13039/501100000274, Adopting an integrative approach, by combining studies of cardiovascular function with those at cellular and molecular levels, this study investigated whether maternal treatment with melatonin protects against programmed cardiovascular dysfunction in the offspring using an established rodent model of hypoxic pregnancy. Wistar rats were divided into normoxic (N) or hypoxic (H, 10% O2) pregnancy ± melatonin (M) treatment (5 μg·ml−1.day−1) in the maternal drinking water. Hypoxia ± melatonin treatment was from day 15–20 of gestation (term is ca. 22 days). To control for possible effects of maternal hypoxia‐induced reductions in maternal food intake, additional dams underwent pregnancy under normoxic conditions but were pair‐fed (PF) to the daily amount consumed by hypoxic dams from day 15 of gestation. In one cohort of animals from each experimental group (N, NM, H, HM, PF, PFM), measurements were made at the end of gestation. In another, following delivery of the offspring, investigations were made at adulthood. In both fetal and adult offspring, fixed aorta and hearts were studied stereologically and frozen hearts were processed for molecular studies. In adult offspring, mesenteric vessels were isolated and vascular reactivity determined by in‐vitro wire myography. Melatonin treatment during normoxic, hypoxic or pair‐fed pregnancy elevated circulating plasma melatonin in the pregnant dam and fetus. Relative to normoxic pregnancy, hypoxic pregnancy increased fetal haematocrit, promoted asymmetric fetal growth restriction and resulted in accelerated postnatal catch‐up growth. Whilst fetal offspring of hypoxic pregnancy showed aortic wall thickening, adult offspring of hypoxic pregnancy showed dilated cardiomyopathy. Similarly, whilst cardiac protein expression of eNOS was downregulated in the fetal heart, eNOS protein expression was elevated in the heart of adult offspring of hypoxic pregnancy. Adult offspring of hypoxic pregnancy further showed enhanced mesenteric vasoconstrictor reactivity to phenylephrine and the thromboxane mimetic U46619. The effects of hypoxic pregnancy on cardiovascular remodelling and function in the fetal and adult offspring were independent of hypoxia‐induced reductions in maternal food intake. Conversely, the effects of hypoxic pregnancy on fetal and postanal growth were similar in pair‐fed pregnancies. Whilst maternal treatment of normoxic or pair‐fed pregnancies with melatonin on the offspring cardiovascular system was unremarkable, treatment of hypoxic pregnancies with melatonin in doses lower than those recommended for overcoming jet lag in humans enhanced fetal cardiac eNOS expression and prevented all alterations in cardiovascular structure and function in fetal and adult offspring. Therefore, the data support that melatonin is a potential therapeutic target for clinical intervention against developmental origins of cardiovascular dysfunction in pregnancy complicated by chronic fetal hypoxia.

Coral bleaching represents the most serious threat to contemporary coral reefs. In response, focus is being laid on understanding the cellular processes involved in the response of corals to the environmental stresses and the molecular mechanisms that determine the bleaching patterns. In the present study, a component of the cellular stress response such as the expression of the heat shock proteins (Hsps) was analyzed following the coral bleaching event which occurred in the central Red Sea (Saudi Arabia) in 2015. During this event, corals of different species, growth forms and sites showed variable bleaching susceptibility. In particular, we investigated the expression of Hsp70, Hsp60 and Hsp32 in both healthy and bleached colonies belonging to four different coral species (Goniopora lobata, Porites lobata, Seriatopora hystrix and Stylophora pistillata), in order to explore the intra- and inter-specific modulation of these biomarkers as well as the existence of spatial patterns of Hsp expression. In healthy colonies, the level of all the biomarkers was significantly different among the different species, although within each species it remained similar regardless of the distance from the shore. All the coral species showed a significant modulation of the Hsp expression in response to bleaching, whose typology and amplitude were species-specific. In all the species, Hsp70 and Hsp60 showed a coordinated dual expression, which, in response to bleaching resulted in an up-regulation in G. lobata and P. lobata and in a down-regulation in S. hystrix and S. pistillata. Hsp32 was up-regulated in all four species following bleaching, indicative of elevated oxidative stress. Overall, the protein expression profiles of each species contribute to assess the role of Hsps in regulating the susceptibility to thermal stresses of the various coral taxa of the Red Sea.

In photosynthesis, the efficiency with which a photogenerated exciton reaches the reaction center is dictated by chromophore energies and the arrangement of chromophores in the supercomplex. Here, we explore the interplay between the arrangement of light-harvesting antennae and the efficiency of exciton transport in purple bacterial photosynthesis. Using a Miller-Abrahams-based exciton hopping model, we compare different arrangements of light-harvesting proteins on the intracytoplasmic membrane. We find that arrangements with aggregated LH1s have a higher efficiency than arrangements with randomly distributed LH1s in a wide range of physiological light fluences. This effect is robust to the introduction of defects on the intracytoplasmic membrane. Our result explains the absence of species with aggregated LH1 arrangements in low-light niches and the large increase seen in the expression of LH1 dimer complexes in high fluences. We suggest that the effect seen in our study is an adaptive strategy toward solar light fluence across different purple bacterial species.

Systematic pre-participation screening of subjects practicing sports activity has the potential to identify athletes at risk of sudden cardiac death. However, limited evidence are present concerning the yield of echocardiography as a second-line exam in athletes with abnormal pre-participation screening.Consecutive athletes were screened (2011-2017) in a community-based sports medicine center in Tuscany, with familial history, physical examination and ECG. Patients with abnormal/1 borderline ECG findings, symptoms/signs of cardiovascular diseases, cardiovascular risk factors or family history of juvenile/genetic cardiac disease underwent echocardiography.A total of 30109 athletes (age 21 [15;31]) were evaluated. Of these, 6234 (21%) were aged 8-11 years, 18309 (61%) 12-18 years, 4442 (15%) 19-35 years, 1124 (4%)35 years. A total of 2569 (9%) athletes were addressed to echocardiography. Referral rates increased significantly with age (5% in preadolescents to 38% in master athletes, P 0.01). Subclinical heart diseases were found in 290/30109 (0.8%) and were common35 years (135/1124, 11%), but rare at 19-35 years (91/4442, 2%), very rare18 years (64/24 543, 0.2%; P 0.01). Seventy-four (0.3%) athletes were disqualified because of the structural alterations identified, 29 (0.1%) with cardiac structural diseases at risk for sudden death.Italian community-based pre-participation screening showed an age-dependent yield, with a three-fold increase in referral in athletes 35 years. Subclinical structural abnormalities potentially predisposing to sudden death were rare (0.01%), mostly in post-pubertal and senior athletes. Age-specific pre-participation screening protocols may help optimize resources and improve specificity.

Background Accurate diagnosis of peanut allergy is a significant clinical challenge. Here, a novel diagnostic blood test using the peanut bead-based epitope assay ("peanut BBEA") was developed utilizing the LEAP cohort and then validated using two independent cohorts. Methods The development of the peanut BBEA diagnostic test followed the National Academy of Medicine's established guidelines with discovery performed on 133 subjects from the non-interventional arm of the LEAP trial and an independent validation performed on 82 subjects from the CoFAR2 and 84 subjects from the POISED study. All samples were analyzed using the peanut BBEA methodology, which measures levels of IgE to two Ara h 2 sequential (linear) epitopes and compares their combination to a threshold pre-specified in the model development phase. When a patient has an inconclusive outcome by skin prick testing (or sIgE), IgE antibody levels to this combination of two epitopes can distinguish whether the patient is "Allergic" or "Not Allergic." Diagnoses of peanut allergy in all subjects were confirmed by double-blind placebo-controlled food challenge and subjects' ages were 7-55 years. Results In the validation using CoFAR2 and POISED cohorts, the peanut BBEA diagnostic test correctly diagnosed 93% of the subjects, with a sensitivity of 92%, specificity of 94%, a positive predictive value of 91%, and negative predictive value of 95%. Conclusions In validation of the peanut BBEA diagnostic test, the overall accuracy was found to be superior to existing diagnostic tests for peanut allergy including skin prick testing, peanut sIgE, and peanut component sIgE testing.

Curcumin-containing nanocapsule powder formulations have not been used in ruminant feed to date, despite the fact that curcumin is known to be a functional food additive. The objective of this study was to determine whether ethyl polymethacrylate (Eudragit L-100) nanocapsules loaded with curcumin (N-CU) would improve health and growth of lambs. Thirty-two male Lacaune lambs (body weight [BW] = 16 ± 0.99 kg; 45 d of age) were randomly assigned to 1 of 4 treatments: T0, T1, T2 and T4, representing supplementation of curcumin at 0, 1, 2, and 4 mg/kg concentrate, respectively. The animals in each treatment were allocated in 4 pens of 2 lambs each (8 lambs per treatment). The experiment lasted 17 d, with samples and measurements collected on d 0, 7, 12, and 17. The T2 lambs had greater average daily gain than T0 lambs. Regression analysis showed that the ideal dose of N-CU to enhance weight gain was 1.89 mg/kg concentrate. There were significant interactions (P

A diverse collection of Juglans species and hybrids with potential to serve as walnut rootstocks was evaluated to identify resistance to Phytophthora cinnamomi, a destructive pathogen affecting commercial production of Persian walnuts worldwide. A total of 35 Juglans genotypes, comprising hybrids and Juglans microcarpa, plus three Juglans regia genotypes as comparison, were inoculated during two seasons (spring and autumn) for 3 years, with two isolates of P. cinnamomi. Inoculations were carried out on excised shoots of the different genotypes by replacing a bark disk with a P. cinnamomi culture plug. After incubation, necrosis length caused by the pathogen was measured. Data were collected and statistically analyzed with generalized linear mixed models. This work pointed out a seasonal influence on some Juglans genotype response to P. cinnamomi: both hybrids and J. microcarpa groups were significantly less sensitive in autumn compared with spring (P = 0.0006), thus this condition must be considered when selecting Juglans for resistance to this pathogen. Three genotypes of J. regia, known for its susceptibility to P. cinnamomi, were used as comparison. Results show good levels of resistance to P. cinnamomi in J. microcarpa genotypes, confirming literature results. Among J. microcarpa genotypes, the Jmi03 is quite promising for its in vitro propagation. A number of Juglans hybrids, H5/18, 7/28,8/29, 10/43, and 6/22, showed significantly higher levels of resistance to P. cinnamomi, compared with susceptible J. regia genotypes. Evaluation, in naturally infected fields, of clonal genotypes and seedling-progenies of resistant genotypes, grafted with commercial walnut varieties, is currently under way and will provide additional information for successful usage.

BACKGROUND: Multifood Oral Immunotherapy (mOIT) with adjunctive anti-IgE (omalizumab, Xolair(®)) treatment affords safe, effective, and rapid desensitization to multiple foods, although the specific immune mechanisms mediating this desensitization remain to be fully elucidated. METHODS: Participants in our phase 2 mOIT trial (NCT02643862) received omalizumab from baseline to week 16 and mOIT from week 8 to week 36. We compared the immune profile of PBMCs and plasma taken at baseline, week 8 and week 36 using high-dimensional mass cytometry, component-resolved diagnostics, the indirect basophil activation test, and Luminex. RESULTS: We found (i) decreased frequency of IL4(+) peanut-reactive CD4(+) T cells and a marked downregulation of GPR15 expression and CXCR3 frequency among γδ and CD8(+) T cell subsets at week 8 during the initial, omalizumab-alone induction phase; (ii) significant upregulation of the skin-homing receptor CCR4 in peanut-reactive CD4(+) T and Th2 effector memory (EM) cells and of cutaneous lymphocyte-associated antigen (CLA) in peanut-reactive CD8(+) T and CD8(+) EM cells (iii) downregulation of CD86 expression among antigen-presenting cell subsets; and (iv) reduction in pro-inflammatory cytokines, notably IL-17, at week 36 post-OIT. We also observed significant attenuation of the Th2 phenotype post-OIT, defined by downregulation of IL-4 peanut-reactive T cells and OX40 in Th2EM cells, increased allergen component-specific IgG(4)/IgE ratio, and decreased allergen-driven activation of indirectly sensitized basophils. CONCLUSIONS: This exploratory study provides novel comprehensive insight into the immune underpinnings of desensitization through omalizumab-facilitated mOIT. Moreover, this study provides encouraging results to support the complex immune changes that can be induced by OIT.

Material for the special course: Diet composition and impacts of invasive round gobies at the East and West of Bornholm

Background & aims: Genome-wide association studies in primary biliary cholangitis (PBC) have failed to find X chromosome (chrX) variants associated with the disease. Here, we specifically explore the chrX contribution to PBC, a sexually dimorphic complex autoimmune disease. Methods: We performed a chrX-wide association study, including genotype data from 5 genome-wide association studies (from Italy, United Kingdom, Canada, China, and Japan; 5244 case patients and 11,875 control individuals). Results: Single-marker association analyses found approximately 100 loci displaying P < 5 × 10-4, with the most significant being a signal within the OTUD5 gene (rs3027490; P = 4.80 × 10-6; odds ratio [OR], 1.39; 95% confidence interval [CI], 1.028-1.88; Japanese cohort). Although the transethnic meta-analysis evidenced only a suggestive signal (rs2239452, mapping within the PIM2 gene; OR, 1.17; 95% CI, 1.09-1.26; P = 9.93 × 10-8), the population-specific meta-analysis showed a genome-wide significant locus in East Asian individuals pointing to the same region (rs7059064, mapping within the GRIPAP1 gene; P = 6.2 × 10-9; OR, 1.33; 95% CI, 1.21-1.46). Indeed, rs7059064 tags a unique linkage disequilibrium block including 7 genes: TIMM17B, PQBP1, PIM2, SLC35A2, OTUD5, KCND1, and GRIPAP1, as well as a superenhancer (GH0XJ048933 within OTUD5) targeting all these genes. GH0XJ048933 is also predicted to target FOXP3, the main T-regulatory cell lineage specification factor. Consistently, OTUD5 and FOXP3 RNA levels were up-regulated in PBC case patients (1.75- and 1.64-fold, respectively). Conclusions: This work represents the first comprehensive study, to our knowledge, of the chrX contribution to the genetics of an autoimmune liver disease and shows a novel PBC-related genome-wide significant locus.

Summary Plants undergo several developmental transitions during their life cycle. In grapevine, a perennial woody fruit crop, the transition from vegetative/green‐to‐mature/woody growth involves transcriptomic reprogramming orchestrated by a small group of genes encoding regulators, but the underlying molecular mechanisms are not fully understood.We investigated the function of the transcriptional regulator VviNAC33 by generating and characterizing transgenic overexpressing grapevine lines and a chimeric repressor, and by exploring its putative targets through a DNA affinity purification sequencing (DAP‐seq) approach combined with transcriptomic data.We demonstrated that VviNAC33 induces leaf de‐greening, inhibits organ growth and directly activates the expression of STAY‐GREEN PROTEIN 1 (SGR1), which is involved in Chl and photosystem degradation, and AUTOPHAGY 8f (ATG8f), which is involved in the maturation of autophagosomes. Furthermore, we show that VviNAC33 directly inhibits AUXIN EFFLUX FACILITATOR PIN1, RopGEF1 and ATP SYNTHASE GAMMA CHAIN 1T (ATPC1), which are involved in photosystem II integrity and activity.Our results show that VviNAC33 plays a major role in terminating photosynthetic activity and organ growth as part of a regulatory network governing the vegetative‐to‐mature phase transition., See also the Commentary on this article by Kelly & Allan, 231: 505–507.

Antibody-Mediated Rejection (AMR) due to donor-specific antibodies (DSA) is associated with poor outcomes after lung transplantation. Currently, there are no guidelines regarding the selection of treatment protocols. We studied how DSA characteristics including titers, C1q, and mean fluorescence intensity (MFI) values in undiluted and diluted sera may predict a response to therapeutic plasma exchange (TPE) and inform patient prognosis after treatment. Among 357 patients consecutively transplanted without detectable pre-existing DSAs between 01/01/16 and 12/31/18, 10 patients were treated with a standardized protocol of five TPE sessions with IVIG. Based on DSA characteristics after treatment, all patients were divided into three groups as responders, partial responders, and nonresponders. Kaplan-Meier Survival analyses showed a statistically significant difference in patient survival between those groups (P = 0.0104). Statistical analyses showed that MFI in pre-TPE 1:16 diluted sera was predictive of a response to standardized protocol (R2 = 0.9182) and patient survival (P = 0.0098). Patients predicted to be nonresponders who underwent treatment with a more aggressive protocol of eight TPE sessions with IVIG and bortezomib showed improvements in treatment response (P = 0.0074) and patient survival (P = 0.0253). Dilutions may guide clinicians as to which patients would be expected to respond to a standards protocol or require more aggressive treatment.

The aim of this work was to reinforce actions tending to reduce Shiga toxin-producing Escherichia coli (STEC) in beef products from an Argentinean commercial abattoir implementing Hazard Analysis and Critical Control Point (HACCP) practices. An environmental stx map was built with 421 environmental samples from the slaughter, quartering, cool chamber and deboning sectors (February-May 2013). For stx determination, 125 carcass and 572 anatomical cut samples were used. Based on the environmental stx mapping results, improvement actions were designed and implemented (June and July 2013). After implementing improvement actions, 160 carcass and 477 anatomical cut samples were collected to identify stx and verify the impact of improvement actions (August-December 2013). Our results showed stx-positivity in pre-operational (10.1%) and operational (15.5%) environmental samples and in carcass and beef cut samples before (4.8 and 10.1%; p = 0.144) and after (1.2 and 4.8%; p = 0.0448) implementing improvement actions, respectively. Although improvement actions reduced stx in beef cuts, it is difficult to implement and sustain a system based on stx zero-tolerance only by reinforcing Good Manufacturing Practices, Sanitation Standard Operating Procedures and HACCP practices. The application of combined intervention strategies to reduce STEC in carcasses and beef cuts should be therefore considered.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was isolated from the oro/pharyngeal swabs of two Italian COVID-19 patients, physicians in a COVID-19 division hospital, with different courses of the disease. The complete genome sequences show that the two isolates belong to the B1.1 lineage, but contain a nucleotide mutation in the ORF6, leading to a stop codon and to the deletion of 6 amino acids in the C terminus. This deletion was unique, compared to the currently available sequences deposited in the GISAID and GenBank database. It did not affect the in vitro viral replication, neither the neutralizing activities of the patients' antibodies. Based on homology analysis with other Coronaviruses, the two isolated lacked the ORF6 aminoacidic portion responsible for the inhibition of the antiviral Interferon (IFN)-based host response. IFN seems to have a dual role of in SARS-CoV-2 infected patients: not only antiviral activity, but also a detrimental role in case of excessive production. A deletion in the SARS-CoV-2 ORF6 protein might have a specific, still unknown role in the viral pathogenesis.

[Image: see text] Asparagine deprivation by l-asparaginase (L-ASNase) is an effective therapeutic strategy in acute lymphoblastic leukemia, with resistance occurring due to upregulation of ASNS, the only human enzyme synthetizing asparagine (Annu. Rev. Biochem.2006, 75 (1), 629–654). l-Asparaginase efficacy in solid tumors is limited by dose-related toxicities (OncoTargets and Therapy 2017, pp 1413–1422). Large-scale loss of function genetic in vitro screens identified ASNS as a cancer dependency in several solid malignancies (Cell2017, 170 (3), 564–576.e16. Cell2017, 170 (3), 577–592.e10). Here we evaluate the therapeutic potential of targeting ASNS in melanoma cells. While we confirm in vitro dependency on ASNS silencing, this is largely dispensable for in vivo tumor growth, even in the face of asparagine deprivation, prompting us to characterize such a resistance mechanism to devise novel therapeutic strategies. Using ex vivo quantitative proteome and transcriptome profiling, we characterize the compensatory mechanism elicited by ASNS knockout melanoma cells allowing their survival. Mechanistically, a genome-wide CRISPR screen revealed that such a resistance mechanism is elicited by a dual axis: GCN2-ATF4 aimed at restoring amino acid levels and MAPK-BCLXL to promote survival. Importantly, pharmacological inhibition of such nodes synergizes with l-asparaginase-mediated asparagine deprivation in ASNS deficient cells suggesting novel potential therapeutic combinations in melanoma.

Adeno-associated virus (AAV) is a small, non-enveloped virus used as vector in gene therapy, mainly produced in human cells and in baculovirus systems. Intense studies on these platforms led to the production of vectors with titers between 103 and 105 viral genomes (vg) per cells. In spite of this, vector yields need to be improved to satisfy the high product demands of clinical trials and future commercialization. Our studies and those of other groups have explored the possibility to exploit the yeast Saccharomyces cerevisiae to produce rAAV. We previously demonstrated that yeast supports AAV genome replication and capsid assembly. The purpose of this study was to evaluate the quality of the encapsidated AAV DNA. Here, we report the construction of a yeast strain expressing Rep68/40 from an integrated copy of the Rep gene under the control of the yeast constitutive ADH promoter and Capsid proteins from the Cap gene under the control of an inducible GAL promoter. Our results indicate that a portion of AAV particles generated by this system contains encapsidated AAV DNA. However, the majority of encapsidated DNA consists of fragmented regions of the transgene cassette, with ITRs being the most represented sequences. Altogether, these data indicate that, in yeast, encapsidation occurs with low efficiency and that rAAVs resemble pseudo-vectors that are present in clinical-grade rAAV preparations.

Large dams disrupt river ecosystems, causing negative effects on fish assemblages. Few studies, however, have investigated how Neotropical fish diversity responds to river regulation in longer temporal scales. In this study, we used a multi-metric approach to investigated temporal changes in taxonomic and functional fish diversity over 15 years-period following the construction of a large dam in the Upper Tocantins River, Amazon. We analyzed biological and environmental data collected in the impounded area before (2000–2002) and after (2003–2014) river regulation. Diversity metrics responded differently to the impoundment. Some metrics were not affected, as they showed little variation over years, such as fish abundance, species, and trait richness. Other metrics changed significantly between periods, such as taxonomic composition, temporal beta diversity, and trait abundance. Species with traits that are sensitive to hydrological alterations declined in the impoundment, while sedentary species were favored. Assemblage trajectories were variable, but community structure developed into a different state in the impoundment. Landscape variables explained changes in fish dissimilarity over time. In general, our study supports the notion that fish diversity in Amazonian rivers is vulnerable to river damming, but also indicates that different metrics show particular responses to this disturbance.

Background Scalp Eschar and Neck LymphAdenopathy after Tick bite is a zoonotic non-pathogen-specific disease most commonly due to Rickettsia slovaca and Rickettsia raoultii. Diagnosis is mostly based only on epidemiological and clinical findings, without serological or molecular corroboration. We presented a clinical case in which diagnosis was supported by entomological identification and by R. slovaca DNA amplifications from the tick vector. Case presentation A 6-year-old child presented with asthenia, scalp eschar and supraclavicular and lateral-cervical lymphadenopathy. Scalp Eschar and Neck LymphAdenopathy After Tick bite syndrome following a Dermacentor marginatus bite was diagnosed. Serological test on serum revealed an IgG titer of 1:1024 against spotted fever group rickettsiae, polymerase chain reaction assays on tick identified Rickettsia slovaca. Patient was successfully treated with doxycycline for 10 days. Conclusions A multidisciplinary approach including epidemiological information, clinical evaluations, entomological identification and molecular investigations on tick, enabled proper diagnosis and therapy.

Aloe ferox Mill is widely used as a traditional herbal medicine for the treatment of a broad spectrum of illnesses given its laxative, anti-inflammatory, bitter tonic, anti-oxidant, antimicrobial and anti-cancer properties. Using the in vivo alkaline comet assay in animals (OECD 489), this study investigated the potential in vivo genotoxicity of dried Aloe ferox juice at dose levels of 500, 1000, and 2000 mg/kg/day in mice. Aloe ferox showed no genotoxic activity in preparations of single cells from the colon of the treated Hsd:ICR (CD-1) male mice. No statistically significant increase in DNA migration over the negative control was observed by analysis of variance for both comet parameters, tail moment and tail intensity, apart from the positive control ethyl methanesulphonate that induced clear and statistically significant increases in DNA migration parameters over the concurrent controls. The new reported scientific evidence unequivocally demonstrates that dried Aloe ferox juice containing hydroxyanthracene derivatives does not induce DNA damage in preparations of single cells from colon in in vivo comet genotoxicity studies. This suggests that the hyperplastic changes and mucosal hyperplasia observed after long-term administration of Aloe vera non-decolourised whole leaf extract may be attributed to an epigenetic effect of the material under investigation.

Non-expressor of pathogenesis-related genes 1 (NPR1) is a key regulator of plant innate immunity and systemic disease resistance. The model for NPR1 function is based on experimental evidence obtained largely from dicots; however, this model does not fit all aspects of Poaceae family, which includes major crops such as wheat, rice and barley. In addition, there is little scientific data on NPR1's role in mutualistic symbioses. We assessed barley (Hordeum vulgare) HvNPR1 requirement during the establishment of mutualistic symbiosis between barley and beneficial Alphaproteobacterium Rhizobium radiobacter F4 (RrF4). Upon RrF4 root-inoculation, barley NPR1-knockdown (KD-hvnpr1) plants lost the typical spatiotemporal colonization pattern and supported less bacterial multiplication. Following RrF4 colonization, expression of salicylic acid marker genes were strongly enhanced in wild-type roots; whereas in comparison, KD-hvnpr1 roots exhibited little to no induction. Both basal and RrF4-induced root-initiated systemic resistance against virulent Blumeria graminis were impaired in leaves of KD-hvnpr1. Besides these immune-related differences, KD-hvnpr1 plants displayed higher root and shoot biomass than WT. However, RrF4-mediated growth promotion was largely compromised in KD-hvnpr1. Our results demonstrate a critical role for HvNPR1 in establishing a mutualistic symbiosis between a beneficial bacterium and a cereal crop.

A cook's mate working in an Austrian restaurant reported acutely occurring urticarial skin lesions after processing and cooking squid. The prick-to-prick test with squid showed a ++ positive urticarial reaction. Elevated specific IgE antibody levels to squid, shrimp, and house dust mites as well as to tropomyosin from shrimp and house dust mite could be detected in the ImmunoCAP. By means of immunoblot and ELISA, a reaction to squid extract as well as increased IgE antibody levels to squid and tropomyosin from squid could be detected. The patient was diagnosed with a clinically and occupationally relevant type I allergy to squid with cross-reaction to tropomyosin of other invertebrates and therefore recognized as an occupational disease.