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SIGNIFICANCE Currently 10-15% of couples in reproductive age face infertility issues. More importantly, male factor contributes to 50% of these cases (either alone or in combination with female causes). Amongst various reasons, impaired sperm function is the main cause for male infertility. Furthermore, mitochondrial dysfunction and oxidative stress due to increased ROS production, particularly of mitochondrial origin, are believed to be main contributors. Recent Advances: Mitochondrial dysfunction, particularly due to increased ROS production, has often been linked to impaired sperm function/quality. For decades different methods and approaches have been developed to assess mitochondrial features that might correlate with sperm functionality. This connection is now completely accepted, with mitochondrial functionality assessment used more commonly as a readout of sperm functionality. More recently, mitochondria-targeted compounds are on the frontline for both assessment as well as therapeutic approaches. CRITICAL ISSUES In this review we summarize the current methods for assessing key mitochondrial parameters known to reflect sperm quality as well as therapeutic strategies using mitochondrial-targeted antioxidants aiming to improve sperm function in various situations, particularly after sperm cryopreservation. FUTURE DIRECTIONS Although more systematic research is needed, mitochondrial-targeted compounds definitely represent a promising tool to assess as well as to protect and improve sperm function.

The invasive Vespa velutina has been widely referred as an effective predator of honeybees. Despite the potential risk to pollination services provision and honey production, there is no accurate quantification and assessment of its real consequences for honeybees. To date, the identification of the honeybee and other insects in the diet of V. velutina has been investigated by direct observation of adult foraging or examination of food pellets. To overcome these limitations, in this study we used a DNA metabarcoding approach to evaluate the usefulness of different types of sample (jaws and stomachs collected from workers and larval faecal pellets taken from the hornet comb) to investigate the predation of V. velutina upon honeybees, and potentially on other insects. Honeybee DNA was identified in all types of samples, but larval faecal pellets retrieved the higher number of reads of honeybee DNA and the largest diversity at all taxonomic levels. Over all samples we could identify 4 orders, 9 families, 6 genera and 1 species of prey. We estimate that collecting 6 workers is sufficient to identify honeybee predation by a colony using worker’s jaws. Stomachs were the least useful sample type to detect honeybee DNA. The presence of honeybee DNA in all analysed colonies irrespective of collection site, and the variety of insect orders detected in the diet support current concerns over the acknowledged negative impact of V. velutina on managed honeybees and its potential threat to pollination services provision.

Funder: Kay Kendall Leukaemia Fund (KKLF); doi: https://doi.org/10.13039/501100000402, Funder: EC | EC Seventh Framework Programm | FP7 People: Marie-Curie Actions (FP7-PEOPLE - Specific Programme "People" Implementing the Seventh Framework Programme of the European Community for Research, Technological Development and Demonstration Activities (2007 to 2013)); doi: https://doi.org/10.13039/100011264; Grant(s): Mitobiopath-705560, Many cellular processes, including ribosome biogenesis, are regulated through post-transcriptional RNA modifications. Here, a genome-wide analysis of the human mitochondrial transcriptome shows that 2'-O-methylation is limited to residues of the mitoribosomal large subunit (mtLSU) 16S mt-rRNA, introduced by MRM1, MRM2 and MRM3, with the modifications installed by the latter two proteins being interdependent. MRM2 controls mitochondrial respiration by regulating mitoribosome biogenesis. In its absence, mtLSU particles (visualized by cryo-EM at the resolution of 2.6 ��) present disordered RNA domains, partial occupancy of bL36m and bound MALSU1:L0R8F8:mtACP anti-association module, allowing five mtLSU biogenesis intermediates with different intersubunit interface configurations to be placed along the assembly pathway. However, mitoribosome biogenesis does not depend on the methyltransferase activity of MRM2. Disruption of the MRM2 Drosophila melanogaster orthologue leads to mitochondria-related developmental arrest. This work identifies a key checkpoint during mtLSU assembly, essential to maintain mitochondrial homeostasis.

Wetland communities are shaped by high levels of stress, disturbance and competition. Using South African palmiet wetlands as a case study (Prionium serratum dominated valley-bottom wetlands), we explore whether autogenic or allogenic succession is the dominant process driving community dynamics in valley-bottom wetlands in drylands. Several wetland rehabilitation programmes in South Africa use the dominant wetland species palmiet (P. serratum) as a pioneer to facilitate recolonization. However research is needed on palmiet wetland dynamics and formation to guide these restoration efforts. We explore vegetation patterns by analyzing which environmental parameters drive dominance of palmiet, resulting in the characteristic patchiness of palmiet wetlands, and which plant functional traits account for this. In 20 plots from three palmiet wetlands distributed across the Cape Floristic Region of South Africa, key soil, groundwater and vegetation parameters, as well as community composition were measured. Twenty-two dominant species were selected and 13 functional traits measured. Soil pH and relative groundwater depth were the main environmental parameters driving community assembly in palmiet wetlands. Palmiet-dominated communities were characterized by greater stem diameter, leaf length–width ratio, leaf area and cellulose and lignin concentration compared to fynbos communities. These traits suggest adaptations to disturbances such as fires (thicker stems) and floods (long, thin leaves, flexible shoots and thicker stems). We propose three hypotheses of palmiet wetland development which shed light on palmiet wetland restoration and highlight gaps for future research.

Influenza NS1 is a promising anti-influenza target, considering its conserved and druggable structure, and key function in influenza replication and pathogenesis. Notwithstanding, target identification and validation, strengthened by experimental data, are lacking. Here, we further explored our previously designed structure-based antiviral rationale directed to highly conserved druggable NS1 regions across a broad spectrum of influenza A viruses. We aimed to identify NS1-mutated viruses exhibiting a reduced growth phenotype and/or an altered cell apoptosis profile. We found that NS1 mutations Y171A, K175A (consensus druggable pocket 1), W102A (consensus druggable pocket 3), Q121A and G184P (multiple consensus druggable pockets) - located at hot spots amenable for pharmacological modulation - significantly impaired A(H1N1)pdm09 virus replication, in vitro. This is the first time that NS1-K175A, -W102A, and -Q121A mutations are characterized. Our map-and-mutate strategy provides the basis to establish the NS1 as a promising target using a rationale with a higher resilience to resistance development. Highlights: Structure-based anti-influenza strategies that facilitate the rapid identification and validation of robust targets are lacking; We have previously identified and characterized 3 NS1 conserved druggable pockets - amenable to pharmacological modulation; NS1 mutations Y171A, K175A (CDP1), W102A (CDP3), Q121A and G184P (Multi-CDPs) impaired A(H1N1)pdm09 virus replication, in vitro; The new NS1 hot spots: NS1–K175, -W102, and -Q121 were characterized for the first time, using the A(H1N1)pdm09 virus; This map-and-mutate antiviral rationale can be promptly applied to other proteins of (re)emergent viruses. This work was funded by the FCT – Fundação para a Ciência e a Tecnologia, I.P. (project number POCI-01 0145-FEDER-032572 | PTDC/QUI-OUT/32572/2017 and PTDC/SAU-INF/30729/2017); and sup ported by the PhD grant PD/BD/128402/2017 from FCT PhD Pro gramme in Medicines and Pharmaceutical Innovation (i3DU) to JTL Ph. D. candidate. info:eu-repo/semantics/publishedVersion

Reactive oxygen/nitrogen species (ROS/RNS) are produced physiologically by several mechanisms, especially during the inflammatory response. However, their overproduction can lead to the evolution of conditions known as oxidative/nitrosative stress, resulting in the establishment of chronic inflammatory diseases. Chalcones are considered as a class of flavonoids having the molecular pattern 1,3-diaryl-2-propen-1-one. In the last few years, the antioxidant property of chalcones has been extensively studied, mainly due to their ability to inhibit the production or scavenging ROS/RNS. The antioxidant activity of chalcones, focusing on the production of ROS/RNS during the inflammatory response, is demonstrated and discussed in the present review. This literature revision was based on the modulatory effects of chalcones against different enzymes, such as superoxide dismutase (SOD), nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, catalase (CAT), myeloperoxidase (MPO) and inducible nitric oxide synthase (iNOS), and, in the scavenging of ROS/RNS. Whenever possible, the structure-activity relationship (SAR) was established. Through the analysis accomplished in this review, it can be observed that the presence of substituents, e.g. hydroxyl, methoxyl, prenyl, and halogen atoms in the chalcones scaffold, often occurs and can improve their modulatory activities, namely, in the production of ROS/RNS during the inflammatory response.

a Universidade Regional do Cariri, URCA, Cel Ant^onio Luis, 1161, 63105-000, Pimenta, Crato, CE, Brazil b Universidade Federal do Paran a, UFPR, XV de Novembro, 1299, 80.060-000, Centro, Curitiba, PR, Brazil c Universidade Regional de Blumenau, FURB, Ant^onio da Veiga, 140, 89030-903, Itoupava Seca, Blumenau, SC, Brazil d Laborat orio de investiga¸c~ao em Gen etica e Hematologia Translacional, Instituto Gon¸calo Moniz (IGM), Funda¸c~ao Oswaldo Cruz (FIOCRUZ), Salvador, BA 40296- 710, Brazil e Centro Universit ario Doutor Le~ao Sampaio (Unile~ao), Av. Maria Leticia Leite Pereira s/n, Lagoa Seca, Juazeiro do Norte, Brazil

© 2022 Prudêncio et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society. This article, published in RNA, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/., Widespread cotranscriptional splicing has been demonstrated from yeast to human. However, most studies to date addressing the kinetics of splicing relative to transcription used either Saccharomyces cerevisiae or metazoan cultured cell lines. Here, we adapted native elongating transcript sequencing technology (NET-seq) to measure cotranscriptional splicing dynamics during the early developmental stages of Drosophila melanogaster embryos. Our results reveal the position of RNA polymerase II (Pol II) when both canonical and recursive splicing occur. We found heterogeneity in splicing dynamics, with some RNAs spliced immediately after intron transcription, whereas for other transcripts no splicing was observed over the first 100 nt of the downstream exon. Introns that show splicing completion before Pol II has reached the end of the downstream exon are necessarily intron-defined. We studied the splicing dynamics of both nascent pre-mRNAs transcribed in the early embryo, which have few and short introns, as well as pre-mRNAs transcribed later in embryonic development, which contain multiple long introns. As expected, we found a relationship between the proportion of spliced reads and intron size. However, intron definition was observed at all intron sizes. We further observed that genes transcribed in the early embryo tend to be isolated in the genome whereas genes transcribed later are often overlapped by a neighboring convergent gene. In isolated genes, transcription termination occurred soon after the polyadenylation site, while in overlapped genes, Pol II persisted associated with the DNA template after cleavage and polyadenylation of the nascent transcript. Taken together, our data unravel novel dynamic features of Pol II transcription and splicing in the developing Drosophila embryo., This work was supported by funding to M.C.-F. (Fundação para a Ciência e Tecnologia, FCT/Ministério da Ciência, Tecnologia e Ensino Superior - Fundos do Orçamento de Estado [UIDB/50005/2020], and FCT/ FEDER/POR Lisboa 2020, Programa Operacional Regional de Lisboa PORTUGAL 2020, grant LISBOA-01-0145-FEDER-016394) and to R.G.M (FCT grant PTDC/BIA-BID/28441/2017). P.P. was a recipient of an FCT fellowship (SFRH/BD/109689/2015). R.S. was a recipient of an EMBO Long-Term Fellowship (EMBO ALTF 101-2019). This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreements no. 842695 (Marie Skłodowska- Curie Actions) and no. 857119 (RiboMed)

Purpose To study the immunization status and IgM and IgG antibody behavior against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in an unvaccinated population of Northeast Portugal (including RT-PCR diagnosed and undiagnosed individuals). Methods Application of a clinical-epidemiological survey, and analysis of IgM and IgG SARS-COV-2 antibodies (against N core protein) in 362 participants that voluntarily sought the laboratory for testing. Results At the time of the analysis, 31.7% (n = 114) of the study population had a previous SARS-CoV-2 diagnosis, 48.3% of which were asymptomatic, and 71.9% IgG seropositive. Of these, 83.3% and 60% were, respectively, IgM and IgG seropositive within 2 weeks after the initial diagnosis. Both antibodies peaked in the 3rd week post diagnosis, with titers decreasing over the following weeks, until a state of seronegativity was achieved after the 6th week for IgM, and the 21st for IgG. Symptomatic patients showed higher IgM and IgG values, when compared to asymptomatic ones. Fever, the most reported symptom, was found to be positively associated with IgM values. Ages of ≤ 18-year-old and ≥ 65-year-old exhibited the highest median values for both IgM and IgG, with the former being statistically significant. In the undiagnosed group, 13.9% and 11.1% were seropositive for IgM and IgG, respectively. Conclusion IgM and IgG displayed a similar initial increase (within 1/2 weeks), with IgG having a significant decrease after the 21st week post-diagnosis, translating a loss of immunity at this point. The youngest and oldest symptomatic age groups were found to be the highest responders. Antibody assays enabled the identification of previously undiagnosed participants.

BRI2 is a type II transmembrane protein ubiquitously expressed whose physiological function remains poorly understood. Although several recent important advances have substantially impacted on our understanding of BRI2 biology and function, providing valuable information for further studies on BRI2. These findings have contributed to a better understanding of BRI2 biology and the underlying signaling pathways involved. In turn, these might provide novel insights with respect to neurodegeneration processes inherent to BRI2-related pathologies, namely Familial British and Danish dementias, Alzheimer's disease, ITM2B-related retinal dystrophy, and multiple sclerosis. In this review, we provided a state-of-the-art outline of BRI2 biology, both in physiological and pathological conditions, and discuss the proposed molecular underlying mechanisms. Overall, the BRI2 knowledge here reviewed is of extreme importance and may contribute to propose BRI2 and/or BRI2 proteolytic fragments as novel therapeutic targets for neurodegenerative diseases, such as Alzheimer's disease.

ACKNOWLEDGEMENTS : This study was partially supported by the European Maritime and Fisheries Fund MAR2020 project “VALOREJET: Valorização de espécies rejeitadas e de baixo valor commercial,” MAR-01.03.01-FEAMP0003, and by Fundação para a Ciência e Tecnologia through research contracts attributed to Vera Sequeira (CEECIND/02705/2017) and Ana Rita Vieira (CEECIND/01528/2017) and strategic project UID/MAR/04292/2019. The depletion and overexploitation of several fish stock demands for a valorisation of non-target and discarded species. Nonetheless, such species are often poorly studied, and information on their biological parameters must be gathered for effective population management. For 1 year, the reproductive strategy of the piper gurnard Trigla lyra and the red gurnard Chelidonichthys cuculus was studied by monthly samples obtained from commercial boats operating on western Portuguese coast. Both species showed a biased sex ratio towards females, especially for larger length classes. Length at first maturity could be estimated only for red gurnard (22.1 and 19.9 cm for females and males, respectively) because all piper gurnard individuals caught were mature. Piper gurnard showed determinate fecundity and a short spawning season, from November to February with a peak in January, whereas red gurnard showed indeterminate fecundity and a wide spawning season, from late December to May. The relative annual fecundity estimated for red gurnard (1893 ± 728 oocytes x g-1 eviscerated weight [EW]) was higher than the one estimated for piper gurnard (1018 ± 250 oocytes x g-1 EW). Although important information for understanding the species dynamics is presented in this study, additional information on other life-history parameters and of species landings is required. info:eu-repo/semantics/publishedVersion

Females from genus Culicoides are responsible for transmitting important pathogenic agents to animals and humans, including bluetongue, Schmallenberg and African horse sickness viruses and filarids. Species identification in laboratory is usually performed by visualization of their wing pattern and/or using other morphological characters. During the National Entomological Surveillance Program for Bluetongue Disease performed in Portugal (2005–2013) several Culicoides midges were captured near domestic ruminants and, more recently, others were collected in a zoo from the Metropolitan Area of Lisbon near sylvatic animals (2018–2019). From a total of 5,364 analysed specimens, some morphological alterations were observed in 154 specimens (2.9 %) from three different Culicoides spp. (Culicoides obsoletus, Culicoides scoticus and Culicoides dewulfi) on palpi (e.g. defective sensorial pits), antennae (e.g. antennae with defective or fused articles), abdomen (e.g. abdomen with aberrant structures) and spermathecae (e.g. specimens with one and four spermathecae). These types of anatomical alterations probably occur due to genetic or morphogenetic malformation. Also, they are compatible with midge survival and thus maintained in adult specimens. The report of different types of anatomical anomalies is important, since these morphological alterations can lead to incorrect species identification during sample analysis. Thus, this work gives essential information concerning some morphological anomalies that can be observed in Culicoides biting midges, being their first reference in C. scoticus and C. dewulfi species.

Background Plasma cell neoplasms are characterized by the proliferation of a single clone of plasma cells with production of a monoclonal immunoglobulin. They can manifest as a single lesion (plasmacytoma) or as multiple lesions (multiple myeloma). Methods Paraffin-embedded tissue blocks of patients microscopically diagnosed with plasma cell neoplasms in the jaws were retrieved from five pathology files. Data including clinical, radiographic, microscopic and immunohistochemical findings, treatment employed, and follow-up status were retrieved from the pathology reports. Results Fifty-two cases were retrieved (mean age: 59.4 years) without sex predilection. The mandible was the most affected site (67.3%), usually associated with pain and/or paresthesia (53.8%). Lesions in other bones besides the jaws were reported for 24 patients (46.2%). Radiographically, tumors usually presented as poorly defined osteolytic lesions with unilocular or multilocular images, while microscopy revealed diffuse proliferation of neoplastic plasma cells with nuclear displacement and abundant eosinophilic cytoplasm. Two cases were classified as anaplastic, and amyloid deposits were found in two other cases. Immunohistochemistry was positive for plasma cell markers and negative for CD20 and CD3, and monoclonality for kappa light chain predominated. The overall survival rate after 5-years of follow-up was 26.6%. Conclusion Plasma cell neoplasms are aggressive tumors with a poor prognosis and involvement of the jaws may be the first complaint of the patient. Thus, oral pathologists, head and neck surgeons and dentists should be aware of their clinical, radiographic and microscopic manifestations.

Made available in DSpace on 2022-04-29T08:29:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) We evaluated the spatiotemporal distribution of Limnoperna fortunei larvae, its abundance in relation to other zooplanktonic organisms (copepoda and cladocerans), and its macrofouling on net cages in the Chavantes Reservoir, Brazil. We also estimated its growth rate under this condition and limnological variables in the water column. Zooplankton densities were positively related to water temperature, and copepoda was the most abundant group. L. fortunei showed the highest density in November and March (18.565 to 30.014 larvae/m3) and the lowest from June to August (1.587 to 876 larvae/m3). Population parameters of L. fortunei, as derived from von Bertalanffy seasonal growth function, indicated fast annual growth (K = 1.22) in length in the first, second and third years of life at 21, 28, and 30 mm, respectively. We estimate that 62.920 juveniles/m2 will be recruited annually in fish farming, mainly in the reproductive period, which should be the focus of prevention methods. Our work is the first report to associate the abundance of L. fortunei larvae in the water column with the recruitment of juveniles. Instituto de Pesca Agência Paulista de Tecnologia dos Agronegócios Postgraduate Program Centro de Aquicultura Universidade Estadual Paulista Postgraduate Program Instituto de Pesca Agência Paulista de Tecnologia dos Agronegócios Instituto de Biofísica Carlos Chagas Filho Universidade Federal do Rio de Janeiro Departamento de Ecología Genética y Evolución Facultad de Ciencias Exactas y Naturales Universidad de Buenos Aires Postgraduate Program Centro de Aquicultura Universidade Estadual Paulista

Background A plasmid-mediated mechanism of bacterial resistance to polymyxin is a serious threat to public health worldwide. The present study aimed to determine the occurrence of plasmid-mediated colistin resistance genes and to conduct the molecular characterization of mcr-positive Escherichia coli strains isolated from Polish poultry. Methods In this study, 318 E. coli strains were characterized by the prevalence of mcr1–mcr5 genes, antimicrobial susceptibility testing by minimal inhibitory concentration method, the presence of antimicrobial resistance genes was screened by PCR, and the biofilm formation ability was tested using the crystal violet staining method. Genetic relatedness of mcr-1-positive E. coli strains was evaluated by multilocus sequence typing method. Results Among the 318 E. coli isolates, 17 (5.35%) harbored the mcr-1 gene. High antimicrobial resistance rates were observed for ampicillin (100%), tetracycline (88.24%), and chloramphenicol (82.35%). All mcr-1-positive E. coli strains were multidrug-resistant, and as many as 88.24% of the isolates contained the blaTEM gene, tetracycline (tetA and tetB), and sulfonamide (sul1, sul2, and sul3) resistance genes. Additionally, 41.18% of multidrug-resistant, mcr-1-positive E. coli isolates were moderate biofilm producers, while the rest of the strains showed weak biofilm production. Nine different sequence types were identified, and the dominant ST was ST93 (29.41%), followed by ST117 (17.65%), ST156 (11.76%), ST 8979 (11.76%), ST744 (5.88%), and ST10 (5.88%). Moreover, the new ST was identified in this study. Conclusions Our results showed a low occurrence of mcr-1-positive E. coli strains isolated from Polish poultry; however, all the isolated strains were resistant to multiple antimicrobial agents and were able to form biofilms at low or medium level.

Introduction: The species Acinetobacter baumannii, within its genus, is the most important for epidemiological surveillance because it is associated, in humans, with different types of infections. Objectives: To evaluate the occurrence and resistance profile of Acinetobacter baumannii in biological samples of patients attended at an oncological hospital. Methods: Descriptive, retrospective and transversal study, carried out from the results of cultures of hospitalized patients and from the outpatient clinic of an oncological hospital, in the period from January 2017 to December 2018. Results: 1,258 positive cultures were recorded in 2017 and 1,729 in 2018. Within this, were obtained 5% of isolates of Acinetobacter baumannii in the first year of analysis and 8% in the second. The topographic site of highest isolation of A. baumannii, was the respiratory tract. The highest rates of occurrence were in older patients. The most frequent hospital sector of the bacteria were the Intensive Care Units. Acinetobacter baumannii showed lower resistance to colistin and tigecycline in both years of the study. The variation of the Minimum Inhibitory Concentration was more evident in the antibacterial drugs colistin. Conclusion: The results show that Acinetobacter baumannii is an important pathogen that causes nosocomial infections, commonly isolated from the respiratory tract of patients hospitalized at Intensive Care Units which presents a high resistance profile to most antibacterial classes.

Context Cesarean section (CS), breastfeeding, and geographic location can influence the infant microbiota. Objective In this systematic review, evidence of the association between mode of delivery and infant gut microbiota up to 6 months of age was evaluated, as was the role of breastfeeding in this association, according to PRISMA guidelines. Data Source The Pubmed, Web of Science, Scopus, Embase, Medical Database, and Open Grey databases were searched. Data Extraction A total of 31 observational studies with ≥2 infant stool collections up to the sixth month of age and a comparison of gut microbiota between CS and vaginal delivery (VD) were included. Data Analysis Infants born by CS had a lower abundance of Bifidobacterium and Bacteroides spp. at almost all points up to age 6 months. Populations of Lactobacillus, Bifidobacterium longum, Bifidobacterium catenulatum, and Escherichia coli were reduced in infants delivered by CS. Infants born by CS and exclusively breastfed had greater similarity with the microbiota of infants born by VD. Conclusions Species of Bifidobacterium and Bacteroides are potentially reduced in infants born by CS. Geographic location influenced bacterial colonization. Systematic Review Registration PROSPERO registration no. 42017071285

The CADM family of proteins consists of four neuronal specific adhesion molecules (CADM1, CADM2, CADM3 and CADM4) that mediate the direct contact and interaction between axons and glia. In the peripheral nerve, axon-Schwann cell interaction is essential for the structural organization of myelinated fibres and is primarily mediated by the binding of CADM3, expressed in axons, to CADM4, expressed by myelinating Schwann cells. We have identified—by whole exome sequencing—three unrelated families, including one de novo patient, with axonal Charcot-Marie-Tooth disease (CMT2) sharing the same private variant in CADM3, Tyr172Cys. This variant is absent in 230 000 control chromosomes from gnomAD and predicted to be pathogenic. Most CADM3 patients share a similar phenotype consisting of autosomal dominant CMT2 with marked upper limb involvement. High resolution mass spectrometry analysis detected a newly created disulphide bond in the mutant CADM3 potentially modifying the native protein conformation. Our data support a retention of the mutant protein in the endoplasmic reticulum and reduced cell surface expression in vitro. Stochastic optical reconstruction microscopy imaging revealed decreased co-localization of the mutant with CADM4 at intercellular contact sites. Mice carrying the corresponding human mutation (Cadm3Y170C) showed reduced expression of the mutant protein in axons. Cadm3Y170C mice showed normal nerve conduction and myelin morphology, but exhibited abnormal axonal organization, including abnormal distribution of Kv1.2 channels and Caspr along myelinated axons. Our findings indicate the involvement of abnormal axon-glia interaction as a disease-causing mechanism in CMT patients with CADM3 mutations.

Hippoboscid flies, also known as louse flies, are obligate blood-feeders ectoparasites of birds and mammals. By studying louse fly parasites of four Passeriformes species, Eurasian blackbird (Turdus merula), Eurasian blackcap (Sylvia atricapilla), common chaffinch (Fringilla coelebs) and European robin (Erithacus rubecula), with dissimilar time of colonization of Azores islands, we tested whether: (i) island host populations have lower parasite richness than the mainland one; (ii) island host populations undergo higher parasite prevalence, mean intensities and mean abundance than the mainland one; (iii) island parasite diversity are composed exclusively by specific parasites and (iv) parasite richness is positively correlated with the island area and proximity to the continent. For these purposes, 775 birds were sampled for presence of louse flies, by modified fumigation chamber method, from Azores Islands (São Miguel, Terceira and Flores) and Portugal mainland. Insular and mainland parasite assemblages were statistically compared. We record for the first time to Azores, Ornithomya fringillina and Icosta minor from Portugal mainland. Louse flies had highest prevalence and abundance from Azores Islands compared to those observed in mainland birds, especially blackbirds. The insular parasite diversity of Azores blackbirds, blackcaps and chaffinches was richer than the one observed in mainland population. None of the hippoboscid flies observed on the islands and mainland were host-specific. Thus, our findings provide an upgrade of parasite island syndromes knowledge, in the context of the ectoparasites, namely to the hippoboscid flies case., Graphical abstract Image 1, Highlights • We investigate the louse flies of four Passeriformes species from Macaronesia. • Louse flies of Azores blackbirds had highest prevalence than in mainland birds. • Insular parasite diversity was richer than the one observed in mainland population. • Hippoboscid flies observed on the islands and mainland were not host-specific. • Ornithomya fringillina recorded for the first time to Azores.

Menstrual symptoms are commonly cited barriers to physical activity in women. Delay or avoidance of menstruation through hormonal contraceptives may mitigate those barriers. Our purpose is to identify the uterine bleeding patterns of Portuguese athletes, their contraceptive choices and the impact of contraceptive methods in sports performance. We aim also to compare the outcomes between users and non-users of contraception.Observational, descriptive and comparative study based on a customised self-assessment online survey aimed at Portuguese athletes. Athletes had to be at least 16 years old and competing at a national, international or professional level.A total of 256 athletes from 18 modalities were eligible. The mean age was 29 years and the mean age of menarche was 12.8 years. Monthly bleeding pattern was prevailing and 50% presented dysmenorrhoea. More than 85% experience fluctuations in sports performance throughout the month. The majority (71.5%) used contraceptive methods. Only 16% believed that contraception decreased sports performance. Contraceptive users exhibit a lower mean body mass index, blood loss and percentage of dysmenorrhoea than the non-users.Most athletes had the perception that performance varies throughout the month. The majority of participants do not report amenorrhoea and have monthly menses. The use of contraceptive methods is reportedly high, benefit cycle control and apparently without any physical inconvenience.

Awareness of the physiological changes that occur when animals are subjected to climatic changes that are considered stressful is essential to maintain animal welfare and to be able to exploit their reproductive potential efficiently and rationally. The present study was carried out to evaluate climatic variables’ influence on physiological parameters, and Murrah buffalo ejaculates reared in a humid tropical climate in the Amazon. The immediate analyzes pertinent to the physical and morphological characteristics of the ejaculates were carried out and corresponded in the rainy season (RS) volume of 3.4±2.0mL; the mass activity of 4.4±0.5; motility of 80.4±5.6%; vigor of 4.4±0.4; concentration of 657,300±237,865.1 x 106sptz/mL; major defects of 9.0±2.6%; minor defects of 11.2±3.9%; total defects 20.2±5.3% and sperm plasma membrane integrity (SPMI) 84.8±5.6%, whereas in the non-rainy season (nRS), the results were 4.0±2.1mL; the mass activity of 3.0±1.0; motility of 56.2±13.4%; vigor of 3.0±1.0; concentration of 586,000±291,925.9 x 106sptz/mL; major defects of 20.8±9.9%; minor defects of 27.5±6.3%; total defects 48.3±9.3% and SPMI of 57.9±12.4%. Furthermore, a statistical difference (P

The study of Diptera at the scene of a crime can provide essential information for the interpretation of evidence. Phylogeographic reconstruction could help differentiate haplotypes of a dipteran species in a geographical area, clarifying, for example, the details of a possible relocation of a corpse. In addition, inferring the ancestral areas of distribution helps to understand the current status of the species and its biogeographic history. One of the most important species in forensic entomology is Calliphora vicina Rovineau-Desvoidy, 1830 (Diptera: Calliphoridae). The aim of this work is to increase our knowledge of this species in the Iberian Peninsula using 464 specimens from Spain and Portugal. These samples were identified using morphological keys and by molecular methods using fragments of the cox1, 16S and ITS2 genes. The phylogeographic history of these populations was inferred from haplotype networks and the reconstruction of ancestral areas of distribution. The molecular results corroborated the morphological identifications of the samples. Phylogeographic networks showed no geographical structure, as haplotypes are shared among almost all populations. reconstruct ancestral state in phylogenies analyses showed a high rate of movement among populations, possibly related to human activity. These results suggest that this species had a very rapid and recent spatial and demographic expansion throughout the Iberian Peninsula.

Background/aim p16 and PTEN are tumor suppressor genes. Loss of these molecules in oral squamous cell carcinoma (OSCC) has been studied worldwide. In this study, we explored whether p16 cooperates with inactive PTEN during the pathogenesis of OSCC, especially in regard to tumor aggressiveness and proliferation. Materials and methods Immunocytochemistry and western blot analysis were used to examine the levels of p16 and PTEN. Sequencing analysis was performed to identify mutations in the PTEN gene and HPV infection. Fluorescence in situ hybridization was used to examine the presence of the PTEN locus. Results PTEN analysis showed high positivity in T4 samples. HPV-positive tumors correlated with tabagism, tumor size 3 and 4, disease stages 3 and 4, presence of lymph node metastasis (N1) and poor differentiation. Immunoexpression of p16 was strongly correlated with the presence of HPV. Conclusion PTEN demonstrated a higher reactivity in advanced disease stages and p16 was strongly associated with HPV. Viral presence decreases tumor aggressiveness. Patients with advanced stage lesions demonstrated lower survival rate.

Aiming to valorise the Atlantic Rainforest biodiversity in Santa Catarina, the chemical characterisation of the essential oils (EOs) from leaves of Vernonanthura montevidensis (Spreng.) H. Rob. is described for the first time. Fresh leaves collected in the year 2014 and 2015, were submitted to hydrodistillation to give pale blue EOs in yields of 0.21 and 0.19%, respectively. The EOs were characterised by GC-MS and GC-FID semi- and quantitative methods. The monoterpene β-pinene was the major constituent in both samples reaching a maximum of 26.3%. The monoterpene α-pinene and the sesquiterpene β-caryophyllene, were also among the major constituents in both samples. By means of the extracted ion chromatogram procedure, it was possible to detect chamazulene, which was associated with the pale blue colour of the essential oils. In the in vitro antimollicute assays, the essential oil was moderately active against Mycoplasma genitalium and M. pneumoniae with MIC values of 250 µg mL−1.

The presence of Sophonia orientalis (Hemiptera: Cicadellidae) in mainland Portugal is reported here for the first time. Previous reports of S. orientalis in mainland Europe include only three detections in the Iberian Peninsula (Gibraltar, Andalusia and Catalonia). Species identification was confirmed both by morphological analysis of male genitalia and molecular analysis of mitochondrial cytochrome c oxidase subunit I gene sequence. Forty specimens of S. orientalis were found associated to three crops (orange orchards, peach orchards and vineyards) from distinct regions across Portugal. The wide distribution of plots where specimens were collected suggests introduction might not be recent, and that this species has successfully established in the territory. Given the known invasive potential of S. orientalis in Hawaii, where it is a pest of multiple crops and a threat to native plant biodiversity, particular attention should be paid to this species in the future.

The acute: chronic workload ratio (ACWR) is an index of the acute workload relative to the cumulative chronic workloads. The monitoring of physical workloads using the ACWR has emerged and been hypothesized as a useful tool for coaches and athletes to optimize performance while aiming to reduce the risk of potentially preventable load-driven injuries. Our goal was to describe characteristics of the ACWR and investigate the association of the ACWR with the risk of time-loss injuries in adult elite team sport athletes. PubMed, EMBASE and grey literature databases; inception to May 2019. Longitudinal studies that assess the relationship of the ACWR and time-loss injury risk in adult professional or elite team sports. We summarized the population characteristics, workload metrics and ACWR calculation methods. For each workload metric, we plotted the risk estimates for the ACWR in isolation, or when combined with chronic workloads. Methodological quality was assessed using a modified version of the Downs and Black scale. Twenty studies comprising 2375 injuries from 1234 athletes (all males and mean age of 24 years) from different sports were included. Internal (65%) and external loads (70%) were collected in more than half of the studies and the session-rating of perceived exertion and total distance were the most commonly collected metrics. The ACWR was commonly calculated using the coupled method (95%), 1:4 weekly blocks (95%) and subsequent week injury lag (80%). There were 14 different binning methods with almost none of the studies using the same binning categories. The majority of studies suggest that athletes are at greater risk of sustaining a time-loss injury when the ACWR is higher relative to a lower or moderate ACWR. The heterogenous methodological approaches not only reflect the wide range of sports studied and the differing demands of these activities, but also limit the strength of recommendations. CRD42017067585.

Emergence and expansion of frequent multidrug-resistant (MDR) major Salmonella clones/serotypes has been a significant threat in the last years. Metal compounds, such as copper, commonly used in animal-production settings, have been pointed out as possible contributors for the selection of such strains/clones. However, the scarcity of studies limits our understanding of the impact of other metal environmental contaminants as arsenic (used in insecticides/herbicides/coccidiostats). We analysed arsenic tolerance (AsT) dispersion by phenotypic and genotypic (PCR/sequencing/I-CeuI/S1/XbaI-PFGE/hybridization) assays among Salmonella with diverse epidemiological and genetic backgrounds. Then, to better understand ars operon genetic contexts, the whole genome of five representative strains was sequenced. We found a high dispersion of ars operons conferring AsT, especially among copper-tolerant and relevant serotypes/clones related to pig-production setting. The acr3-type was found dispersed in the chromosome of diverse serotypes, including the emergent S. Rissen. Conversely, arsBII was almost confined to the MDR ST34 European clone of S. Typhimurium/S. 4,[5],12:i:-, always along with copper/silver tolerance sil + pco clusters in an integrative conjugative element. These data suggest that AsT is an essential adaptive feature for the ecological success of these epidemic clones/serotypes and alerts for global strategies to reduce arsenic-based compounds' impact thus preventing environmental/food contamination with frequent MDR foodborne pathogens.

Limonium mucronatum (L.f.) Chaz. (Plumbaginaceae), a strict endemic of Morocco, has a restricted range in the littoral zone between Rabat and Tan Tan (Draa valley). This coastal region at the junction of Mediterranean, Canary and Saharan influences is of considerable biogeographical interest. However, little information exists on L. mucronatum communities and its relationship with bioclimatology or chromosome polymorphisms, which may have an impact in the context of plant conservation. In this study, we analysed L. mucronatum communities and performed a cytometric and karyological characterization of this rare species. Results showed two plant communities associated with this species related to a latitudinal and bioclimatic gradient along the surveyed region. Our study provided for the first time new data regarding nuclear DNA amount and about the numbers, positions and organization of 45S rDNA loci in L. mucronatum. Remarkably, cytogenetic analyses revealed homogeneous ploidy across all studied populations with all individuals with 2n¼12 chromosomes. The results obtained are discussed in the context of the originality of flora and the threats to L. mucronatum communities, and in the perspective of a conservation strategy for this endemic species info:eu-repo/semantics/publishedVersion

Located on the northern coast of Santa Maria Island (Azores Archipelago, central North Atlantic), the Lagoinhas section preserves a carbonate buildup correlated with Marine Isotope Substage (MIS) 5e, the warmest interval of the Last Interglacial. The buildup is formed mainly by crustose coralline algae (CCA) identified as Spongites sp., and some subordinate crusts of Lithophyllum sp. and Neogoniolithon sp., as well as cf. Titanoderma sp. Extant CCA buildups are not recorded in the archipelago. Herein, we describe in detail the morphological and taphonomical features of the Lagoinhas CCA buildup and interpret the environment in which it grew. Additionally, this buildup is compared with another of similar age, exposed in the Prainha-Praia do Calhau section on the island's opposite southern coast. The hydrodynamic regime appears to play a crucial role in the development of Azorean CCA buildups during the MIS 5e. We thank Direccao Regional da Ciência e Tecnologia (Regional Government of the Azores), FCT (Fundação para a Ciência e a Tecnologia) of the Portuguese Government, and Camara Municipal de Vila do Porto for financial support. We also acknowledge the field assistance of Camara Municipal de Vila do Porto. We are grateful to the organizers and participants of the 16 editions of the International Workshop Palaeontology in Atlantic Islands who helped with fieldwork (2002 2020). A.C.R. was supported by a grant SFRH/BPD/117810/2016 from FCT (Fundação para Ciência e Tecnologia), Portugal. S.P.A. and R.S.R. acknowledge their research contracts (IF/00465/2015 and IF/01641/2015, respectively) funded by Fundação para a Ciência e a Tecnologia (FCT). C.S.M. is benefiting from a PhD grant M3.1.a/F/100/2015 from Fundo Regional para a Ciência e Tecnologia (FRCT). This work was supported by FEDER funds through the Operational Programme for Competitiveness Factors -COMPETE, by Portuguese National Funds through FCT under the projects UID/BIA/50027/2013, UID/BIA/00329/2013-2023, POCI-01-0145-FEDER-006821, PLATMAR (PTDC/GEO-GEO/0051/2014) and under DRCT-M1.1.a/005/Funcionamento-C-/2016 (CIBIO-A) project from FRCT, and by FEDER funds (in 85%) and by funds of the Regional Government of the Azores (15%) trough Programa Operacional Acores 2020, in the scope of the projects "AZORESBIOPORTAL -PORBIOTA": ACORES-01-0145FEDER-000072 and VRPROTO -Virtual Reality PROTOtype: the geological history of "Pedra-que-pica": ACORES-01-0145-FEDER000078. A.U. benefited from additional support by the Jagiellonian University. info:eu-repo/semantics/publishedVersion

Commercial plantations and alien tree invasions often have substantial negative impacts on local biodiversity. The effect of plantations on faunal communities in the fire-adapted fynbos vegetation of the Cape Floristic Region biodiversity hotspot is not yet well quantified. We studied small mammal community structure in alien Pinus radiata plantations and adjacent fynbos regenerating after clear-felling of plantations on the Cape Peninsula, South Africa. Small mammal sampling over 1,800 trap-nights resulted in 480 captures of 345 individuals (excluding recaptures) representing six species. Significantly more species, individuals (12 X) and biomasses (29 X) of small mammals occurred on recovering fynbos sites compared to plantations. This was commensurate with a higher diversity of plant growth forms, vegetation densities and live vegetation biomass. Only one small mammal species, the pygmy mouse (Mus minutoides), was consistently trapped within plantations. Fynbos sites were dominated by three small mammal species that are ecological generalists and early successional pioneer species, rendering the recovering fynbos slightly depauperate in terms of species richness and evenness relative to other studies done in pristine fynbos. We make three recommendations for forestry that would facilitate the restoration of more diverse natural plant communities and progressively more diverse and dynamic small mammal assemblages in a key biodiversity hotspot.

Conjugative plasmids are extrachromosomal mobile genetic elements pervasive among bacteria. Plasmids' acquisition often lowers cells' growth rate, so their ubiquity has been a matter of debate. Chromosomes occasionally mutate, rendering plasmids cost-free. However, these compensatory mutations typically take hundreds of generations to appear after plasmid arrival. By then, it could be too late to compete with fast-growing plasmid-free cells successfully. Moreover, arriving plasmids would have to wait hundreds of generations for compensatory mutations to appear in the chromosome of their new host. We hypothesize that plasmid-donor cells may use the plasmid as a ‘weapon’ to compete with plasmid-free cells, particularly in structured environments. Cells already adapted to plasmids may increase their inclusive fitness through plasmid transfer to impose a cost to nearby plasmid-free cells and increase the replication opportunities of nearby relatives. A mathematical model suggests conditions under which the proposed hypothesis works, and computer simulations tested the long-term plasmid maintenance. Our hypothesis explains the maintenance of conjugative plasmids not coding for beneficial genes. This article is part of the theme issue ‘The secret lives of microbial mobile genetic elements’.

Differences between males and females are usually more subtle in dioecious plants than animals, but strong sexual dimorphism has evolved convergently in the South African Cape plant genus Leucadendron. Such sexual dimorphism in leaf size is expected largely to be due to differential gene expression between the sexes. We compared patterns of gene expression in leaves among 10 Leucadendron species across the genus. Surprisingly, we found no positive association between sexual dimorphism in morphology and the number or the percentage of sex-biased genes (SBGs). Sex bias in most SBGs evolved recently and was species specific. We compared rates of evolutionary change in expression for genes that were sex biased in one species but unbiased in others and found that SBGs evolved faster in expression than unbiased genes. This greater rate of expression evolution of SBGs, also documented in animals, might suggest the possible role of sexual selection in the evolution of gene expression. However, our comparative analysis clearly indicates that the more rapid rate of expression evolution of SBGs predated the origin of bias, and shifts towards bias were depleted in signatures of adaptation. Our results are thus more consistent with the view that sex bias is simply freer to evolve in genes less subject to constraints in expression level.

Description, Hybridization footprint suggests introgression with local wild relatives in Western European grape varieties., Grapevine (Vitis vinifera L.) diversity richness results from a complex domestication history over multiple historical periods. Here, we used whole-genome resequencing to elucidate different aspects of its recent evolutionary history. Our results support a model in which a central domestication event in grapevine was followed by postdomestication hybridization with local wild genotypes, leading to the presence of an introgression signature in modern wine varieties across Western Europe. The strongest signal was associated with a subset of Iberian grapevine varieties showing large introgression tracts. We targeted this study group for further analysis, demonstrating how regions under selection in wild populations from the Iberian Peninsula were preferentially passed on to the cultivated varieties by gene flow. Examination of underlying genes suggests that environmental adaptation played a fundamental role in both the evolution of wild genotypes and the outcome of hybridization with cultivated varieties, supporting a case of adaptive introgression in grapevine.

Although mutations in more than 90 genes are known to cause CMT, the underlying genetic cause of CMT remains unknown in more than 50% of affected individuals. The discovery of additional genes that harbor CMT2-causing mutations increasingly depends on sharing sequence data on a global level. In this way—by combining data from seven countries on four continents—we were able to define mutations in ATP1A1, which encodes the alpha1 subunit of the Na+,K+-ATPase, as a cause of autosomal-dominant CMT2. Seven missense changes were identified that segregated within individual pedigrees: c.143T>G (p.Leu48Arg), c.1775T>C (p.Ile592Thr), c.1789G>A (p.Ala597Thr), c.1801_1802delinsTT (p.Asp601Phe), c.1798C>G (p.Pro600Ala), c.1798C>A (p.Pro600Thr), and c.2432A>C (p.Asp811Ala). Immunostaining peripheral nerve axons localized ATP1A1 to the axolemma of myelinated sensory and motor axons and to Schmidt-Lanterman incisures of myelin sheaths. Two-electrode voltage clamp measurements on Xenopus oocytes demonstrated significant reduction in Na+ current activity in some, but not all, ouabain-insensitive ATP1A1 mutants, suggesting a loss-of-function defect of the Na+,K+ pump. Five mutants fall into a remarkably narrow motif within the helical linker region that couples the nucleotide-binding and phosphorylation domains. These findings identify a CMT pathway and a potential target for therapy development in degenerative diseases of peripheral nerve axons.

Genome instability (GIN) is a main contributing factor to congenital and somatic diseases, but its sporadic occurrence in individual cell cycles makes it difficult to study mechanistically. One profound manifestation of GIN is the formation of micronuclei (MN), the engulfment of chromosomes or chromosome fragments in their own nuclear structures separate from the main nucleus. Here, we developed MN-seq, an approach for sequencing the DNA contained within micronuclei. We applied MN-seq to mice with mutations in Mcm4 and Rad9a, which disrupt DNA replication, repair, and damage responses. Data analysis and simulations show that centromere presence, fragment length, and a heterogenous landscape of chromosomal fragility all contribute to the patterns of DNA present within MN. In particular, we show that long genes, but also gene-poor regions, are associated with chromosome breaks that lead to the enrichment of particular genomic sequences in MN, in a genetic background-specific manner. Finally, we introduce single-cell micronucleus sequencing (scMN-seq), an approach to sequence the DNA present in MN of individual cells. Together, sequencing micronuclei provides a systematic approach for studying GIN and reveals novel molecular associations with chromosome breakage and segregation.

In multifunctional landscapes, diverse communities of flying vertebrate predators provide vital services of insect pest control. In such landscapes, conservation biocontrol should benefit service-providing species to enhance the flow, stability and resilience of pest control services supporting the production of food and fiber. However, this would require identifying key service providers, which may be challenging when multiple predators interact with multiple pests. Here we provide a framework to identify the functional role of individual species to pest control in multifunctional landscapes. First, we used DNA metabarcoding to provide detailed data on pest species predation by diverse predator communities. Then, these data were fed into an extensive network analysis, in which information relevant for conservation biocontrol is gained from parameters describing network structure (e.g., modularity) and species roles in such network (e.g., centrality, specialization). We applied our framework to a Mediterranean landscape, where 19 bat species were found to feed on 132 insect pest species. Metabarcoding data revealed potentially important bats that consumed insect pest species in high frequency and/or diversity. Network analysis showed a modular structure, indicating sets of bat species that are required to regulate specific sets of insect pests. A few generalist bats had particularly important roles, either at network or module levels. Extinction simulations highlighted six bats, including species of conservation concern, which were sufficient to ensure that over three-quarters of the pest species had at least one bat predator. Combining DNA metabarcoding and ecological network analysis provides a valuable framework to identify individual species within diverse predator communities that might have a disproportionate contribution to pest control services in multifunctional landscapes. These species can be regarded as candidate targets for conservation biocontrol, although additional information is needed to evaluate their actual effectiveness in pest regulation.

Cellular proliferation depends on the accurate and timely replication of the genome. Several genetic diseases are caused by mutations in key DNA replication genes; however, it remains unclear whether these genes influence the normal program of DNA replication timing. Similarly, the factors that regulate DNA replication dynamics are poorly understood. To systematically identify trans-acting modulators of replication timing, we profiled replication in 184 cell lines from three cell types, encompassing 60 different gene knockouts or genetic diseases. Through a rigorous approach that considers the background variability of replication timing, we concluded that most samples displayed normal replication timing. However, mutations in two genes showed consistently abnormal replication timing. The first gene was RIF1, a known modulator of replication timing. The second was MCM10, a highly conserved member of the pre-replication complex. MCM10 mutant cells demonstrated replication timing variability comprising 46% of the genome and at different locations than RIF1 knockouts. Replication timing alterations in MCM10-mutant cells was predominantly comprised of replication initiation defects. Taken together, this study demonstrates the remarkable robustness of the human replication timing program and reveals MCM10 as a novel modulator of DNA replication timing.

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Graphical abstract, Highlights • Human SARSr-CoV Spike protein is highly conserved in both S1 and S2 subunits. • The S1-RBD, -SD1; S2-CR, -HR1 and -CH are the most promising druggable S regions. • 28 main consensus druggable pockets were found with high druggabbility score. • New hot spots were identified for hSARSr-CoVs (n = 181) and Beta-CoVs (n = 72). • A structure-based rationale is disclosed for drug design and discovery., There are no approved target therapeutics against SARS-CoV-2 or other beta-CoVs. The beta-CoV Spike protein is a promising target considering the critical role in viral infection and pathogenesis and its surface exposed features. We performed a structure-based strategy targeting highly conserved druggable regions resulting from a comprehensive large-scale sequence analysis and structural characterization of Spike domains across SARSr- and MERSr-CoVs. We have disclosed 28 main consensus druggable pockets within the Spike. The RBD and SD1 (S1 subunit); and the CR, HR1 and CH (S2 subunit) represent the most promising conserved druggable regions. Additionally, we have identified 181 new potential hot spot residues for the hSARSr-CoVs and 72 new hot spot residues for the SARSr- and MERSr-CoVs, which have not been described before in the literature. These sites/residues exhibit advantageous structural features for targeted molecular and pharmacological modulation. This study establishes the Spike as a promising anti-CoV target using an approach with a potential higher resilience to resistance development and directed to a broad spectrum of Beta-CoVs, including the new SARS-CoV-2 responsible for COVID-19. This research also provides a structure-based rationale for the design and discovery of chemical inhibitors, antibodies or other therapeutic modalities successfully targeting the Beta-CoV Spike protein.

The Mediterranean Basin is a global biodiversity hotspot, hosting a number of native species belonging to families that are found almost exclusively in tropical climates. Yet, whether or not these taxa were able to survive in the Mediterranean region during the Quaternary climatic oscillations remains unknown. Focusing on the European free‐tailed bat (Tadarida teniotis), we aimed to (a) identify potential ancient populations and glacial refugia; (b) determine the post‐glacial colonization routes across the Mediterranean; and (c) evaluate current population structure and demography. Mitochondrial and nuclear markers were used to understand T. teniotis evolutionary and demographic history. We show that T. teniotis is likely restricted to the Western Palearctic, with mitochondrial phylogeny suggesting a split between an Anatolian/Middle East clade and a European clade. Nuclear data pointed to three genetic populations, one of which is an isolated and highly differentiated group in the Canary Islands, another distributed across Iberia, Morocco, and France, and a third stretching from Italy to the east, with admixture following a pattern of isolation by distance. Evolutionary and demographic reconstruction supports a pre‐Last Glacial Maximum (LGM) colonization of Italy and the Anatolian/Middle East, while the remaining populations were colonized from Italy after the Younger Dryas. We also found support for demographic expansion following the Iberian colonization. The results show that during the LGM T. teniotis persisted in Mediterranean refugia and has subsequently expanded to its current circum‐Mediterranean range. Our findings raise questions regarding the physiological and ecological traits that enabled species with tropical affinities to survive in colder climates.

Ongoing climate change is expected to cause an increase in temperature and a reduction of precipitation levels in the Mediterranean region, which might cause changes in many species distributions. These effects negatively influence species gene pools, decreasing genetic variability and adaptive potential. Here, we use mitochondrial DNA and RADseq to analyse population genetic structure and genetic diversity of the bumblebee species Bombus terrestris (subspecies Bombus terrestris lusitanicus), in the Iberian Peninsula. Although this subspecies shows a panmictic pattern of population structure across Iberia and beyond, we found differentiation between subspecies B. t. lusitanicus and B. t. africanus, probably caused by the existence of barriers to gene flow between Iberia and North Africa. Furthermore, the results revealed that the Iberian Peninsula harbours a large fraction of B. terrestris intraspecific genetic variation, with the highest number of mitochondrial haplotypes found when compared with any other region in Europe studied so far, suggesting a potential role for the Iberian Peninsula as a glacial refugium. Our findings strengthen the idea that Iberia is a very important source of diversity for the global genetic pool of this species, because rare alleles might play a role in population resilience against human- or climate-mediated changes.

The Antarctic continent is known for its harsh conditions, but it can sustain well-adapted microorganisms that are able to survive in these extreme conditions. Different bacterial phyla are found predominating the soils of Antarctic Peninsula including those of King George Island, which is part of the South Shetlands archipelago. It has been previously demonstrated that Firmicutes make up a substantial proportion of the bacterial communities in rhizosphere soils of the region. However, far less is known about the presence of this group of bacteria in different soils of this island. Eleven soil samples of Admiralty Bay, King George Island (Yellow soil, Copacabana, Demay Point, Domeyko Glacier, Hennequin, Ipanema, Machu Picchu, Arctowski, Smok Beach, Punta Plaza and Vale Ulman) were obtained and analysed using culture-independent methods (clone libraries based on the 16S rRNA encoding gene). In total, 582 clones were obtained and those related to Firmicutes were found in all samples varying from 3% (Vale Ulman) to 62.2% (Machu Picchu). The genus Sporosarcina (61.4%) followed by the genus Bacillus (29.1%) predominated among the Firmicutes-related clones. Non-metric multidimensional scaling analysis (NMDS) showed that potassium and phosphorous concentrations influenced the diversity and relative abundance of the different genera belonging to Firmicutes in Copacabana and Arctowski soils, while soil texture was the main driver in Punta Plaza, Machu Picchu, Yellow soil and Demay Point. This study contributes to our current knowledge of the diversity of Firmicutes in different soils across King George Island, Antarctic Peninsula.

Fossil evidence suggests that scent glands are basal features of Testudines. However, we know little about the structure of these glands in the Brazilian Kinosternidae. In this study, we described the macroscopic anatomy, histology, and histochemistry of the scent glands of three males and three females of Kinosternon scorpioides scorpioides from the Marajó mesoregion, Pará State, Brazil. In all of the specimens analyzed, regardless of sex, we found four scent glands, including two axillary and two inguinal glands that were structurally similar to each other. Each gland consisted of a single holocrine secretory lobule, a large lumen surrounded by relatively thin glandular secretory epithelium, an adjacent narrow layer of loose connective tissue, and a thick layer of skeletal striated muscle tissue surrounded by a serous tunic. The secretory epithelium produced a characteristic malodorous yellowish substance that was passed via a single duct through a bone channel in the bridge connecting the carapace to the plastron and excreted through an outer pore in the plate of each respective gland. Histologically, the secretory epithelium presented cells with two types of secretory vacuoles. Type 1 vacuoles stained red were the largest and most frequently found, and stained positively with Periodic acid-Schiff (PAS), suggesting they contained glycoproteic complexes. Type 2 vacuoles were translucent, smaller in size and fewer in number, and negative for PAS staining. Because they are very primitive structures, scent glands must play important roles in the lives of chelonians, but their real function remains unknown. Several hypotheses suggest that they can act as protection against ectoparasites, as a repellent of predators, in addition to attracting mates and eliciting other pheromonal responses. In this study, all animals reacted by exuding malodorous substances when handled, as a form of defense. However, these are just assumptions that need to be clarified with additional studies on animal behavior. Anat Rec, 303:1489-1500, 2020. © 2019 American Association for Anatomy.