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Severe clinical diseases associated to αCoronavirus (αCoV) infections were recently demonstrated for the first time in humans and a closely related but distinct canine CoV (CCoV) variant was identified in the nasopharyngeal swabs of children with pneumonia hospitalized in Malaysia, in 2017-2018. The complete genome sequence analysis demonstrated that the isolated strain, CCoV-HuPn-2018, was a novel canine-feline-like recombinant virus with a unique nucleoprotein. The occurrence of three human epidemics/pandemic caused by CoVs in the recent years and the detection of CCoV-HuPn-2018, raises questions about the ability of these viruses to overcome species barriers from their reservoirs jumping to humans. Interestingly, in this perspective, it is interesting to consider the report concerning new CCoV strains with a potential dual recombinant origin through partial S-gene exchange with porcine transmissible gastroenteritis virus (TGEV) identified in pups died with acute gastroenteritis in 2009. The significance of the ability of CCoVs to evolve is still unclear, but several questions arisen on the biology of these viruses, focusing important epidemiological outcomes in the field, in terms of both virus evolution and prophylaxis. The new CCoV-Hupn-2018 should lead researchers to pay more attention to the mechanisms of recombination among CoVs, rather than to the onset of variants as a result of mutations, suggesting a continuous monitoring of these viruses and in particular of SARS-CoV-2.

Objectives The main goal of this study was to accurately detect azole resistance in species of the Aspergillus fumigatus complex by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Methods Identification of isolates (n = 868) was done with MALDI-TOF MS using both commercial and in-house libraries. To determine azole susceptibility, the EUCAST E.Def. 9.3.2 method was applied as the reference standard. Identification of resistant isolates was confirmed by DNA sequence analysis. Protein spectra obtained by MALDI-TOF MS were analysed to differentiate species within the A. fumigatus complex and to detect azole-resistant A. fumigatus sensu stricto isolates. Results Correct discrimination of A. fumigatus sensu stricto from cryptic species was accomplished in 100% of the cases applying principal component analysis (PCA) to protein spectra generated by MALDI-TOF MS. Furthermore, a specific peak (4586 m/z) was found to be present only in cryptic species. The application of partial least squares (PLS) discriminant analysis allowed 98.43% (±0.038) discrimination between susceptible and azole-resistant A. fumigatus sensu stricto isolates. Finally, based on PLS and SVM, A. fumigatus sensu stricto isolates with different cyp51A gene mutations were correctly clustered in 91.5% of the cases. Conclusions MALDI-TOF MS combined with peak analysis is a novel tool that allows the differentiation of A. fumigatus sensu stricto from other species within the A. fumigatus complex, as well as the detection of azole-resistant A. fumigatus sensu stricto. Although further studies are still needed, the results reported here show the great potential of MALDI-TOF and machine learning for the rapid detection of azole-resistant Aspergillus fumigatus isolates from clinical origins.

Unresectable neuroendocrine neoplasms (NENs) often poorly respond to standard therapeutic approaches. Alkylating agents, in particular temozolomide, commonly used to treat high-grade brain tumors including glioblastomas, have recently been tested in advanced or metastatic NENs, where they showed promising response rates. In glioblastomas, prediction of response to temozolomide is based on the assessment of the methylation status of the MGMT gene, as its product, O 6-methylguanine-DNA methyltransferase, may counteract the damaging effects of the alkylating agent. However, in NENs, such a biomarker has not been validated yet. Thus, we have investigated MGMT methylation in 42 NENs of different grades and from various sites of origin by two different approaches: in contrast to methylation-specific PCR (MSP), which is commonly used in glioblastoma management, amplicon bisulfite sequencing (ABS) is based on high-resolution, next-generation sequencing and interrogates several additional CpG sites compared to those covered by MSP. Overall, we found MGMT methylation in 74% (31/42) of the NENs investigated. A higher methylation degree was observed in well-differentiated tumors and in tumors originating in the gastrointestinal tract. Comparing MSP and ABS results, we demonstrate that the region analyzed by the MSP test is sufficiently informative of the MGMT methylation status in NENs, suggesting that this predictive parameter could routinely be interrogated also in NENs.

Introduction: The resistance to antibiotics of Helicobacter pylori (H. pylori) is the main factor that affects current therapeutic treatments. The main objective of this study is to describe the pattern of antibiotic resistances in children with an infection due to H. pylori. Patients and methods: An observational, retrospective study was conducted from 2014 to 2019, which included patients between 5 and 17 years old, on whom a gastroscopy, with a gastric biopsy culture positive for H. pylori, and an antibiotic sensitivity study was performed. The antibiotic sensitivity studies were performed using an epsilometer (E-test). The cut-off points to define the resistances were those proposed by the European Committee on Antimicrobial Susceptibility Testing — EUCAST. The eradication study was performed using the 13C-urea breath test or the H. pylori monoclonal test in faeces 6–8 weeks after finalising the treatment. Results: The study included 80 patients (63.8% females), with a mean age of 11.9 years (SD ± 2.7 DS). Over one-third (38.8%) of the patients had received previous treatment for H. pylori. In the endoscopy, peptic ulcer lesions were observed in 10% of patients. More than two-thirds (67.5%) had resistance to at least one drug. 16.3% presented double resistance. The primary resistances were: clarithromycin, 44.9%, metronidazole 16.3%, levofloxacine 7.9%, and amoxicillin 2%. Patients that received treatment according to the new ESPGHAN 2017 guidelines had significantly higher eradication rates compared to those that received treatment according to previous guidelines (80% vs. 55.8%, P = 0.04). Conclusions: The high rate of H. pylori resistances, and as a result, the low eradication rates, are still a very important cause for concern. The first line treatment, when this is indicated must be given following the antibiotic sensitivity studies, and in the cases where these cannot be done or are not available, at least in accordance with the regional resistance rates. The correct application of the new guidelines significantly improves the eradication rate. Resumen: Introducción: Las resistencias antibióticas de Helicobacter pylori (H. pylori) son el principal factor que afecta a la eficacia de los regímenes terapéuticos actuales. El objetivo principal del estudio es describir el patrón de resistencias antibióticas en ni˜nos con infección por H. pylori. Pacientes y métodos: Estudio observacional retrospectivo de 2014 a 2019 en el que se incluyen pacientes entre 5–17 años a los que se realizó gastroscopia, con cultivo de biopsia gástricapositivo para H. pylori y estudio de sensibilidad a antibióticos. Los estudios de sensibilidad antibiótica se realizaron mediante E-test. Los puntos de corte para definir las resistencias fueron los propuestos por el EUCAST. El estudio de erradicación se realizó con test del aliento con urea marcada con C 13 o test monoclonal de antígeno de H. pylori en heces a las 6–8 semanas de finalizar el tratamiento. Resultados: Ochenta pacientes (63,8% mujeres). Media de edad 11,9 años (±2,7 DS). Un 38,8% habían recibido tratamiento previo para H. pylori. Un 10% presentaron en la endoscopia lesiones ulcerosas pépticas. El 67,5% presentaba resistencia al menos a un fármaco. Un 16,3% presentaron doble resistencia. Las resistencias primarias fueron: claritromicina 44,9%, metronidazol 16,3%, levofloxacino 7,9% y amoxicilina 2%. Los pacientes que recibieron tratamiento acorde a las nuevas guías ESPGHAN 2017 presentaron tasas de erradicación significativamente superiores en comparación con los que recibieron tratamiento acorde a las guías previas (80% vs. 55,8% p = 0,04). Conclusiones: La alta tasa de resistencias de H. pylori y, en consecuencia, las bajas tasas de erradicación, siguen siendo una preocupación muy importante. El tratamiento de primera línea, cuando esté indicado debe hacerse guiado por estudios de sensibilidad antibiótica y en los casos en el que no se puedan realizar o no estén disponibles, al menos de acuerdo con las tasas regionales de resistencia. La aplicación correcta de las nuevas guías mejora de forma significativa el nivel de erradicación.

Introduction: There is no consensus on the management of the coronavirus disease (COVID-19) in patients with secondary immunosuppression due to either an underlying hematological disease or to the effects of immunochemotherapy (ICT). Some of them may present persistent infection with multiple relapses of COVID-19, requiring several admissions. This study evaluated the clinical characteristics and outcomes after treatment of 5 patients with follicular lymphoma (FL), previously treated with ICT, who developed several episodes of COVID-19. Methods: We analyzed the clinical evolution and response to treatment with antiviral agent, steroids, and convalescent plasma in 5 patients with FL and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) persistent infection. Reverse transcriptase polymerase chain reaction tests and peripheral blood immunophenotype were performed for all patients. Results: All patients required hospitalization due to pneumonia with severity criteria and were re-admitted after a median of 22 days (13–42) from the previous discharge. They all showed B-cell depletion by immunophenotyping, and no traces of immunoglobulin antibodies against SARS-CoV-2 were detected in any of the cases. The survival rate was 80%. Conclusion: The combination therapy evidenced clinical benefits, demonstrating its capacity to control infection in immunosuppressed FL patients treated with ICT.

Stable isotopes are increasingly being used to unlock the wealth of information contained in specimens preserved in museum collections. However, preservation methods that employ formalin may confound ecological interpretations. To quantify the effects of formalin fixation and subsequent fluid storage in ethanol on the isotopic signatures of small mammal hair, we analyzed δ 13C and δ 15N values from specimens of seven rodent species that were sampled repeatedly both before and after varying lengths of formalin fixation (1–11 days) and ethanol storage (1–6 years). We supplemented these data with a 2-week fixation experiment using deer mice (Peromyscus maniculatus) in which no ethanol storage was employed. As expected, preservation in formalin and ethanol had no discernable effect on δ 15N values. In contrast, specimen δ 13C values decreased in a saturating fashion during formalin fixation and over subsequent years of fluid storage in ethanol. On the basis of models that we fit to these time series, we estimate the long-term effect of fixation and storage on δ 13C values to be −0.92‰ after 4 years. This biologically relevant shift in δ 13C values should be accounted for when inferring the diets of species from fluid-stored museum collections and when comparing across specimens with different preservation histories.

CD36 glycoprotein is a candidate receptor involved in the gustatory detection of lipids and emerging evidence has suggested that genetic variations in CD36 may modulate the oral perception threshold to fatty acids. Here, we analyzed the association of -31118 G > A polymorphism in CD36 gene with nutritional status and preferences for fatty foods in Mexican children. Genotyping of SNP rs1761667 was performed in school-age children (n = 63) in addition to sensory tests evaluating the preference and satisfaction score assigned to oil-based sauces of different fatty acid composition. The G allele was associated with high BMI z-score in children (OR = 2.43, 95% (CI 1.02–5.99); p = 0.02) but CD36 genotypes (AA, GA, and GG) did not show significant association with the preference and satisfaction scores assigned to oil-based sauces. The BMI z-score showed no association with the preference to oil-based sauces; however, children with normal weight gave higher satisfaction scores to sauces with a high content of unsaturated fatty acids than to sauces rich in saturated fatty acids (0.56 ± 1.26 vs. 0.06 ± 1.22; p = 0.02). Therefore, the G allele of -31118 G > A SNP in CD36 gene is associated with overweight and obesity in Mexican children but do not appear to modulate the preferences and satisfaction scores to fat. info:eu-repo/semantics/acceptedVersion

The Mediterranean diet (MD) is becoming a milestone for the prevention of chronic diseases, such as cardiovascular diseases (CVDs), Alzheimer’s and Parkinson’s disease. Ancel Keys in the 1950’s showed a low mortality rate, particularly for coronary heart disease, among people resident in the Mediterranean area. The MD is characterized by the intake of the high amount of vegetables, fruit, and cereals and regular but moderate consumption of wine, fish, and dairy products, while olive oil is the main source of culinary fat. Therefore, it is principally a plant-based diet rich in polyphenols, a heterogeneous category of compounds with different properties and bioavailabilities. Among polyphenols, anthocyanins have been combined into the human food regime for centuries. They have been utilized as traditional herbal remedies for their ability to treat several conditions, as potent anti-oxidants, anti-diabetic and anti-carcinogenic compounds. This review summarizes our knowledge on the health-enhancing component of the anthocyanins-rich diet.

Background & Aims: The ANSWER study reported that long-term albumin administration in patients with cirrhosis and uncomplicated ascites improves survival. During treatment, serum albumin increased within a month and remained stable thereafter. In this post hoc analysis, we aimed to determine whether on-treatment serum albumin levels could guide therapy. Methods: Logistic regression was used to assess the association between baseline serum albumin and mortality, as well as to determine on-treatment factors associated with mortality and to predict the achievement of a given on-treatment serum albumin level. Survival was assessed by Kaplan-Meier estimates and second-order polynomial regression. Patients whose on-treatment serum albumin remained below normal were compared with a subset of patients from the control arm matched by principal score. Results: Baseline serum albumin was closely associated with 18-month mortality in untreated patients; albumin treatment almost effaced this relationship. On-treatment serum albumin and MELD-Na at month 1 were the sole independent variables associated with mortality. Second-order polynomial regression revealed that survival improved in parallel with increased 1-month on-treatment serum albumin. Kaplan-Meier estimations showed that any value of 1-month on-treatment serum albumin (0.1 g/dl intervals) in the range 2.5–4.5 g/dl discriminated patient survival. In the normal range of serum albumin, the best discriminant value was 4.0 g/dl. Compared to untreated patients, survival even improved in patients whose on-treatment serum albumin remained below normal. Conclusion: Baseline serum albumin per se should not guide the decision to start albumin therapy. Conversely, 1-month on-treatment serum albumin levels are strongly associated with outcomes and could guide the use of albumin – 4.0 g/dl being the target threshold. However, even patients whose serum albumin remains below normal benefit from long-term albumin administration. Lay summary: The ANSWER study has shown that long-term albumin administration improves survival and prevents the occurrence of major complications in patients with cirrhosis and ascites. This study shows that the achievement of these beneficial effects is related to a significant increase in serum albumin concentration. Even though the best results follow the achievement of a serum albumin concentration of 4 g/dl, a survival benefit is also achieved in patients who fail to normalise serum albumin.

Familial adenomatous polyposis (FAP) is a rare genetic disorder caused mainly by monoallelic mutations of APC gene. The hereditary breast and ovarian cancer (HBOC) syndrome is an autosomal dominantly inherited disease, which mostly predisposes to breast and ovarian cancers as a result of germline mutations in BRCA1 or BRCA2 genes. In a family, mutations in two cancer susceptibility genes are extremely rare. We studied a family with a case of a 46 years-old woman affected with FAP and ovarian cancer. The patient was affected with profuse FAP since the age of 18 years and a serous ovarian cancer was diagnosed at the age of 45 years. She reported other FAP cases and one case of breast cancer in maternal family. Initially, she was tested for FAP predisposition with mutational analysis of APC gene that revealed the presence of a frameshift mutation, c.3927_3931delAAAGA (p.Glu1309AspfsX4). The presence of ovarian cancer in the patient and of a breast cancer case in the maternal family, suggested an extended analysis to HBOC susceptibility genes that led to the detection of a frameshift mutation, c.3756_3759delGTCT (p.Ser1253Argfs), in BRCA1 gene. The genetic analysis was extended also to family members. The occurrence of the double mutation in APC and BRCA1 genes in the patient was responsible for the onset of FAP and ovarian cancer respectively. The genetic counselling in hereditary cancer with a careful analysis of the pedigree allows identifying the gene to be analyzed. The development of multi-gene panels testing for cancer predisposition, with next generation sequencing (NGS), may reveal mutations in genes of high and moderate penetrance for cancer, although at a low frequency and allows diagnosing a possible double heterozygosity. This enables an adjusted treatment for the affected patient and is critical as it allows initiation of early risk-reducing measures for identified mutation carriers among family members.

Notwithstanding several research efforts in the past years, robust and replicable molecular signatures for autism spectrum disorders from peripheral blood remain elusive. The available literature on blood transcriptome in ASD suggests that through accurate experimental design it is possible to extract important information on the disease pathophysiology at the peripheral level. Here we exploit the availability of a resource for molecular biomarkers in ASD, the Italian Autism Network (ITAN) collection, for the investigation of transcriptomic signatures in ASD based on a discordant sibling pair design. Whole blood samples from 75 discordant sibling pairs selected from the ITAN network where submitted to RNASeq analysis and data analyzed by complementary approaches. Overall, differences in gene expression between affected and unaffected siblings were small. In order to assess the contribution of differences in the relative proportion of blood cells between discordant siblings, we have applied two different cell deconvolution algorithms, showing that the observed molecular signatures mainly reflect changes in peripheral blood immune cell composition, in particular NK cells. The results obtained by the cell deconvolution approach are supported by the analysis performed by WGCNA. Our report describes the largest differential gene expression profiling in peripheral blood of ASD subjects and controls conducted by RNASeq. The observed signatures are consistent with the hypothesis of immune alterations in autism and an increased risk of developing autism in subjects exposed to prenatal infections or stress. Our study also points to a potential role of NMUR1, HMGB3, and PTPRN2 in ASD.

Introduction of alien species leading to unfavorable economic impacts and ecosystem collapse is a well-known threat. The aim was to define if high salinity may be a limiting factor for the survival and spread of the invasive crayfish Procambarus clarkii. Both sexes were exposed to increasing salt concentrations reaching 35.3‰ after 65 days to simulate the natural transition from freshwater into seawater. Higher mortality was recorded for salinity-treated females than for males. Condition indexes gave evidence of minor adverse effects, whereas altered values of oxidative stress biomarkers suggested a perturbation of redox state induced by the exposure conditions. The female survivors showed a strengthening of levels of oxidative stress biomarkers. A moderate oxidative pressure was recorded for gills in both sexes. Nevertheless, the non-indigenous red swamp crayfish has shown great resistance and adaptability to these simulated adverse environmental conditions. Survival of P. clarkii at high salinity may suggest that both sexes can be able to invade estuarine brackish water and lagoons initially exploited as ecological corridors, causing ecological imbalances in transitional ecosystems and in seawater. Moreover, this alien invasive species could be able to descend rivers up to the sea and vice versa to colonize new environments.

Introduction Blood coagulation constituents might exert immunomodulatory functions in the CNS and may trigger neuroinflammation and demyelination. We evaluated whether particular single-nucleotide polymorphisms (SNPs), thought to be involved in fibrinogen-mediated hemostatic pathways, are overrepresented in patients with MS compared with controls. Methods The case–control study consisted of 119 MS patients recruited consecutively at our clinic, and 68 healthy controls. Afterwards, we created a cumulative genetic risk score (CGRS) which included the 5 selected hemostatic risk alleles (Beta-Fibrinogen 455G/A, Glycoprotein IIIa P1A2, Factor V Leiden, Factor V H2R, and Prothrombin 20210G/A). Multivariate ordinal logistic regression and multivariate multinomial logistic regression were applied to evaluate the effect of CGRS on MS susceptibility. Results The FGB 455 G/A and Factor V H1299R variants might be associated with MS status, in the recessive and dominant model, respectively. A cumulative association of the five SNPs investigated with the disease was observed. Discussion We found that MS patients carried more pro-hemostatic variants than healthy controls. An increasing number of unfavorable alleles might increase the likelihood of being in the MS group, in the cumulative analysis. Our findings encourage to evaluating these variants in a larger population-based cohort.

Navua sedge (Cyperus aromaticus) is an aggressive perennial sedge in pastures and crops in the wet tropical regions of Australia and numerous Pacific Island countries and territories. A biological ...

High genetic variation and extensive gene flow may help forest trees with adapting to ongoing climate change, yet the genetic bases underlying their adaptive potential remain largely unknown. We investigated range-wide patterns of potentially adaptive genetic variation in 64 populations of European beech (Fagus sylvatica L.) using 270 SNPs from 139 candidate genes involved either in phenology or in stress responses. We inferred neutral genetic structure and processes (drift and gene flow) and performed differentiation outlier analyses and gene-environment association (GEA) analyses to detect signatures of divergent selection.Beech range-wide genetic structure was consistent with the species’ previously identified postglacial expansion scenario and recolonization routes. Populations showed high diversity and low differentiation along the major expansion routes. A total of 52 loci were found to be putatively under selection and 15 of them turned up in multiple GEA analyses. Temperature and precipitation related variables were equally represented in significant genotype-climate associations. Signatures of divergent selection were detected in the same proportion for stress response and phenology-related genes. The range-wide adaptive genetic structure of beech appears highly integrated, suggesting a balanced contribution of phenology and stress-related genes to local adaptation, and of temperature and precipitation regimes to genetic clines. Our results imply a best-case scenario for the maintenance of high genetic diversity during range shifts in beech (and putatively other forest trees) with a combination of gene flow maintaining within-population neutral diversity and selection maintaining between-population adaptive differentiation.

Catalytic nucleic acids, such as ribozymes, are central to a variety of origin-of-life scenarios. Typically, they require elevated magnesium concentrations for folding and activity, but their function can be inhibited by high concentrations of monovalent salts. Here we show that geologically plausible high-sodium, low-magnesium solutions derived from leaching basalt (rock and remelted glass) inhibit ribozyme catalysis, but that this activity can be rescued by selective magnesium up-concentration by heat flow across rock fissures. In contrast to up-concentration by dehydration or freezing, this system is so far from equilibrium that it can actively alter the Mg:Na salt ratio to an extent that enables key ribozyme activities, such as self-replication and RNA extension, in otherwise challenging solution conditions. The principle demonstrated here is applicable to a broad range of salt concentrations and compositions, and, as such, highly relevant to various origin-of-life scenarios.

Background: Injuries contribute to morbidity and mortality in children. This study was carried out to describe the pattern of childhood injuries and associated risk factors in Dar es Salaam, Tanzania. Methods: This case control study was conducted in six selected health facilities in Dar es Salaam, Tanzania. Data were col- lected using a structured questionnaire. Cases and controls were children below 18 years who had suffered injuries and those without injury associated condition respectively. Results: A total of 492 cases and 492 controls were included in the study, falls (32%), burns (26%), Road Traffic Injuries (14%) and cuts (10%) were the major types of injuries identified. Younger parents/guardians {Adjusted odds ratio (AOR)= 1.4; 95% CI: 1.4 -3.6}, more than six people in the same house (AOR= 1.8; 95% CI: 1.3-2.6), more than three children in the house {AOR= 1.4; 95% CI (1.0-2.0)}, absence of parent/guardian at time of injury occurrence (AOR= 1.6; 95% CI: 1.1-2.3), middle socio-economic (AOR=1.6; 95%CI: 1.1-2.4) and low socio-economic status (AOR= 1.5; 95% CI: 1.0-2.1) were independent risk factors for childhood injury. Conclusion: Falls, burns and road traffic injuries were the main injury types in this study. Inadequate supervision, over- crowding, lower socio-economic status and low maternal age were significant risk factors for childhood injuries. Keywords: Childhood injuries; risk factors; Dar es Salaam; Tanzania.

Plasmacytoid dendritic cells (pDCs) are potent producers of type I IFN (IFN-I) during viral infection and respond to IFN-I in a positive feedback loop that promotes their function. IFN-I shapes dendritic cell responses during helminth infection, impacting their ability to support Th2 responses. However, the role of pDCs in type 2 inflammation is unclear. Previous studies have shown that pDCs are dispensable for hepatic or splenic Th2 responses during the early stages of murine infection with the trematode Schistosoma mansoni at the onset of parasite egg laying. However, during S. mansoni infection, an ongoing Th2 response against mature parasite eggs is required to protect the liver and intestine from acute damage and how pDCs participate in immune responses to eggs and adult worms in various tissues beyond acute infection remains unclear. We now show that pDCs are required for optimal Th2 cytokine production in response to S. mansoni eggs in the intestinal-draining mesenteric lymph nodes throughout infection and for egg-specific IFN-γ at later time points of infection. Further, pDC depletion at chronic stages of infection led to increased hepatic and splenic pathology as well as abrogated Th2 cell cytokine production and activation in the liver. In vitro, mesenteric lymph node pDCs supported Th2 cell responses from infection-experienced CD4+ T cells, a process dependent on pDC IFN-I responsiveness, yet independent of Ag. Together, these data highlight a previously unappreciated role for pDCs and IFN-I in maintaining and reinforcing type 2 immunity in the lymph nodes and inflamed tissue during helminth infection.

Angela J Okragly,1 Katie Brannon Corwin,2 Marikka Elia,3 Dongmei He,3 Oliver Schroeder,3 Qing Zhang,3 Tatiyana Shiyanova,2 Stuart Bright,1 Sarah B Dicker,4 Lukasz Chlewicki,4 Stephanie ME Truhlar,3 Julian Davies,3 Chetan N Patel,2 Robert J Benschop1 1Immunology Research, Eli Lilly and Company, Indianapolis, IN, USA; 2BioTechnology Discovery Research, Eli Lilly and Company, Indianapolis, IN, USA; 3BioTechnology Discovery Research Eli Lilly and Company, San Diego, CA, USA; 4ADME, Eli Lilly and Company, Indianapolis, IN, USACorrespondence: Angela J OkraglyImmunology Research, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN, 46285, USATel +1- 317-276-2839Email okragly_angela@lilly.comBackground: Interleukin-33 (IL-33) is an alarmin that is released following cellular damage, mechanical injury, or necrosis. It is a member of the IL-1 family and binds to a heterodimer receptor consisting of ST2 and IL-1RAP to induce the production of a wide range of cellular mediators, including the type 2 cytokines IL-4, IL-5 and IL-13. This relationship has led to the hypothesis that the IL-33/ST2 pathway is a driver of allergic disease and inhibition of the IL-33 and ST2 association could have therapeutic benefit.Methods: In this paper, we describe the selection of a phage antibody through the ability to bind human IL-33 and block IL-33/ST2 interaction. This hit antibody was then affinity matured by site-directed mutagenesis of the antibody complementarity-determining regions (CDRs). Further characterization of a fully human monoclonal antibody (mAb), torudokimab (LY3375880) included demonstration of human IL-33 neutralization activity in vitro with an NFκB reporter assay and IL-33 induced mast cell cytokine secretion assay, followed by an in vivo IL-33-induced pharmacodynamic inhibition assay in mice that used IL-5 production as the endpoint.Results: Torudokimab is highly specific to IL-33 and does not bind any of the other IL-1 family members. Furthermore, torudokimab binds human and cynomolgus monkey IL-33 with higher affinity than the binding affinity of IL-33 to ST2, but does not bind mouse, rat, or rabbit IL-33. Torudokimab’s half-life in cynomolgous monkey projects monthly dosing in the clinic.Conclusion: Due to torudokimab’s high affinity, its ability to completely neutralize IL-33 activity in vitro and in vivo, and the observed cynomolgus monkey pharmacokinetic properties, this molecule was selected for clinical development.Keywords: IL-33, Th2 immune response, monoclonal antibody

As part of an Integrated Multiple Case Study (Yin, 2013), a method applied for the implementation of a Sustainable Management Model for the Mining-Metallurgical Industry of Mexico, the individual study of the mining analysis unit is presented, which is in the closing stage and belongs to the Au-Ag-Pb-Cu-Zn Mineralization Trend of the national territory. In addition to collaborating with the validation of the Model in field, this study aims to evaluate whether environmental, practices comply with legislative requirements and align with the international suggestions of the UN (2016) through Sustainable Development Goals selected from the agenda 2030. The contribution of this study lies in the importance that is generated from the closure strategy they have followed and how it impacts on the environment, involving ecological and social aspects primarily; the mining unit in the closing stage has been involved in clashes led by radical groups, arguing excessive devastation of important areas, forcing the corporation to execute plans for total closure.

Since 2013, the olive landscapes have gradually degenerated due to the spread and establishment of Xylella fastidiosa subsp. pauca (hereafter Xf) in Apulia, southern Italy. From 2013 to 2019, a total of approximately 54,000 hectares of olive orchards in the south of this region have been seriously damaged, and their restoration will progressively regenerate the economic, social, cultural and environmental nonmarket benefits. Since there is a willingness to restore the affected landscape in the best interest of the local citizens, this research aims to predict their preference heterogeneity and willingness to pay (WTP) to improve this landscape and continue research and experimentation in relation to Olive Quick Decline Syndrome Disease by the bacterium. For this purpose, a choice experiment method is used. The social field survey includes a representative sample of 683 respondents in three major cities (Foggia, Bari and Lecce) of Apulia region. The results reveal that for the local citizens interviewed, the most appreciated olive landscape services are cultural heritage and aesthetic values. In addition, the findings revealed citizens’ positive appreciation of improving the damaged olive landscape, while respondents are not willing to pay a premium for research. The results show that the average value that Apulians are willing to pay for landscape restoration is about 5.7 million of € per year. Further, this research has implications for land use planners in the study area, which faces issues of harmful pathogen management and land revival.

Susceptibility status of Aedes albopictus (Skuse) sampled from residential areas in Interior, Sandakan and Tawau divisions of Sabah, Malaysia, was evaluated based on the WHOrecommended doses of organochlorine and organophosphate larvicides. To determine susceptibility status, larval bioassays were carried out and post 24-hour mortalities based on WHO resistance classifications were adopted. The results demonstrated that Ae. albopictus larvae were resistant toward 5 out of the 8 larvicides tested. Larvae from all populations were resistant against bromophos, fenitrothion, malathion, temephos and dichlorodiphenyltrichloroethane (DDT), with mortalities ranging from 0.00 to 89.33%. Dieldrin, on the other hand, could induce 100.00% mortalities in all populations, followed by fenthion and chlorpyrifos, with mortalities ranging from 97.33 to 100.00% and 81.33 to 100.00% respectively. Despite most populations exhibiting similitude in their resistance status, larvae from Sandakan exhibited the highest resistance level whereas the lowest level was observed in Keningau. In view of the inadequacy of some larvicides in controlling Ae. albopictus in this study, integrated management such as insecticide rotation or combination of interventions is warranted.

Introduction: Monitoring resistance to first line Antiretroviral therapy (ART) is crucial in preventing accumulation of viral mutations following the implementation of the World Health Organization “treat all” initiative. We estimated the rate and predictors of virological treatment failure among adults living with HIV/AIDS in Dar es Salaam, Tanzania. Methodology: A retrospective cohort study involving adults aged 18 and above receiving first line ART in Dar as Salaam between 2016 and 2018 were recruited using multistage random sampling. Clinical and laboratory data were extracted from Care and Treatment Clinic database-2 (CTC2) followed by participant’s interviews. Adjusted Cox-regression modelling was used to determine independent predictors of treatment failure. Results: A total of 340 participants with mean age of 37 were recruited. Overall, 10.59% had virological failure and the rate of failure was 5.24 (95% CI:3.72; 7.27) per 100 person-months at risk with a median failure time of 18 months. Independent predictors of treatment failure were being a male (Adjusted hazard ratio (aHR) 2.78, 95%CI:1.16;6.63), having used treatment for less than two years (aHR, 12.48, 95%CI:3.64-22.71) and co-infection with Tuberculosis (aHR 2.1, 95%CI: 1.0;5.9). Conclusions: HIV virological failure occurs early during treatment in this population. Male clients, co-infected with Tuberculosis were at higher risk of ART failure within two years of treatment. Substantial stride has been made towards the achievement of the last UNAIDS 90 goal but tailored counseling and close monitoring of HIV/TB co-infected male clients following ART initiation could accelerate efforts to close the gap. Further studies on pre-treatment drug resistance mutations are called for.

This study examined some factors associated with the utilization of maternal health care servicesby adolescent mothers (15-19 years) in Tanzania in order to provide advice accordingly. The studyused cross-sectional study of adolescent mothers aged 15-19 years using Demographic HealthSurvey and Malaria indicator Survey 2015/16 data. The dependent variables were number ofantenatal care visits, the place where an adolescent mother delivered and post-natal checkup(adolescent mother’s health checking after being discharged or after a home delivery). Theindependent variables were birth order, education level of a mother, marital status of a mother,media exposure, wealth index, distance to health facility. Multiple binary logistic regression wasused to examine an association between each dependent variable and their respective independentvariables. Data was analyzed using IBM SPSS statistics and STATA. This study used 550adolescent mothers in the analysis. Majority of the adolescent mothers had less than four AntenatalCare (ANC) visits (53.5%), while 68.5% of adolescent mothers delivered at a health facility.Adolescent mothers with two or more children had less odds of having at least four ANCscompared to those with one child, whereas adolescent mothers with at least secondary educationhad greater odds of delivering at a health facility compared to those who had no education.Adolescent mothers who had at least four antenatal care visits and those who are married hadgreater odds of checking their health after being discharged compared to adolescent mothers whohad less than 4 ANCs and single adolescent mothers. It was advised that provision of maternaleducation to young girls on the importance of safe delivery and health checking after delivery isvery important to reduce adolescent maternal morbidity and mortality in the country. Keywords: Adolescent; Maternal Health; Logistic regression; Chi-square

La presente investigación se desarrolló en el municipio de Vallbona de les Monges, Lleida, España, en la Cooperativa L ́Olivera Societat, durante el ciclo vegetativo 2013. Los sistemas de conducción estudiados fueron LYS (constituido por un monoplano ascendente y uno descendente) y monoplano ascendente (MA). Se evaluó la influencia del sistema de conducción sobre el rendimiento y la calidad de mosto de vid (Vitis vinífera L.) Var. “Macabeu”, el comportamiento agronómico, estructura del dosel, potencial hídrico, componentes del rendimiento y calidad del mosto, entre otras variables. Se realizó análisis de varianza de una sola clasificación y la comparación de medias con la prueba de Tukey. Las plantas del sistema LYS presentaron una mayor superficie foliar externa total (SFET) de 18 153.5 m2∙ha-1, un menor número de capas de hojas (NCH, 2.4), mayor exposición de los racimos de uvas que el MA. El porcentaje de brotación fue de 97 %, rendimiento de 31.72 t·ha-1, y 11.6 grado probable de alcohol. Estas variables tuvieron diferencias estadísticas significativas (P

The synthesis of both enantiomers of a key intermediate in the synthesis of halofuginone was accomplished by a Candida antarctica lipase B (CAL-B)-catalyzed kinetic resolution of the corresponding racemate. When the resolution was carried out in the versatile solvent cyclopentyl methyl ether (CPME) using p-chlorophenylbutyrate (PCPB) as the acylating reagent, the highest enantiomeric ratio (E) values were measured, and highly enantioenriched (95% ee) compounds could be obtained in a single iteration. As an example, one of the two enantiomers was used as a starting material to prepare (+)-halofuginone in a three-step procedure.

Oak forests (Fagaceae) are native in Lebanonand occupy the largest areas of approximately 40,000 ha. The most common species are Quercus calliprinos, Q. infectoria, Q. cerris var. pseudo cerris andQ.brantii. Due to climate change and human activities, oak forests have become more vulnerable to native and exotic invasive pests. A total of 26insect species associated with oak trees were recently identified in Lebanon. The most dangerous insect pestisthe gypsy moth Lymantria dispar (Lepidoptera), onQ. calliprinos andQ. Cerris. The oak leafminer Phyllonorycter libanotica (Lepidoptera)and the Eriophidae(Accari) arethe most species recorded on Q. infectoriaand Q.calliprinosfollowed by the giant mealybug Ceroputo pilosellae(Hemiptera) on Q. infectoriaand Q.calliprinosandthe oak mothThaumetopoea sp.(Lepidoptera) on Q. calliprinos and Q. Cerris. Eightnew species were recorded for the first time in Lebanon on oak and are listed in this paper. Fivespecies of Cinipidae (Hymenoptera): Andricus caputmedusae, A. cecconi, A. sternlichti, Plagiotrochus quercusilicisand Neuroterus quercusbaccarum, one speciesof Scolytidae,Xylosandrus compactus(Coleoptera),one species ofKermesidaeKermes echinatus (Hemiptera)and one species of Diaspididae,Koroneaspis aegilopos(Hemiptera).Keywords: Lebanon, oak, forest decline, invasive species, outbreak

A hallmark of meiosis is chromosomal pairing, which requires telomere tethering and rotation on the nuclear envelope through microtubules, driving chromosome homology searches. Telomere pulling toward the centrosome forms the “zygotene chromosomal bouquet.” Here, we identified the “zygotene cilium” in oocytes. This cilium provides a cable system for the bouquet machinery and extends throughout the germline cyst. Using zebrafish mutants and live manipulations, we demonstrate that the cilium anchors the centrosome to counterbalance telomere pulling. The cilium is essential for bouquet and synaptonemal complex formation, oogenesis, ovarian development, and fertility. Thus, a cilium represents a conserved player in zebrafish and mouse meiosis, which sheds light on reproductive aspects in ciliopathies and suggests that cilia can control chromosomal dynamics.

Splenic marginal zone B-cell lymphoma (SMZL) is a heterogeneous clinico-biological entity. The clinical course is variable, multiple genes are mutated with no unifying mechanism, and essential regulatory pathways and surrounding microenvironments are diverse. We sought to clarify the heterogeneity of SMZL by resolving different subgroups and their underlying genomic abnormalities, pathway signatures, and microenvironment compositions to uncover biomarkers and therapeutic vulnerabilities. We studied 303 SMZL spleen samples collected through the IELSG46 multicenter international study (NCT02945319) by using a multiplatform approach. We carried out genetic and phenotypic analyses, defined self-organized signatures, validated the findings in independent primary tumor metadata and determined correlations with outcome data. We identified 2 prominent genetic clusters in SMZL, termed NNK (58% of cases, harboring NF-κB, NOTCH, and KLF2 modules) and DMT (32% of cases, with DNA-damage response, MAPK, and TLR modules). Genetic aberrations in multiple genes as well as cytogenetic and immunogenetic features distinguished NNK- from DMT-SMZLs. These genetic clusters not only have distinct underpinning biology, as judged by differences in gene-expression signatures, but also different outcomes, with inferior survival in NNK-SMZLs. Digital cytometry and in situ profiling segregated 2 basic types of SMZL immune microenvironments termed immune-suppressive SMZL (50% of cases, associated with inflammatory cells and immune checkpoint activation) and immune-silent SMZL (50% of cases, associated with an immune-excluded phenotype) with distinct mutational and clinical connotations. In summary, we propose a nosology of SMZL that can implement its classification and also aid in the development of rationally targeted treatments.

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.

Metabolic pathway regulates cell fate Lineage-specific regulators direct cell fate decisions, but the precise mechanisms are not well known. Using an in vivo chemical suppressor screen of a bloodless zebrafish mutant, Rossmann et al. show that the lineage-specific chromatin factor tif1γ directly regulates mitochondrial genes to drive red blood cell differentiation. Loss of tif1γ reduces coenzyme Q synthesis and function, impeding mitochondrial respiration and leading to epigenetic alterations and repression of erythropoiesis. The loss of blood in the mutant fish can be rescued by the addition of coenzyme Q. This work establishes a mechanism by which a chromatin factor tunes a metabolic pathway in a tissue-specific manner. Science , this issue p. 716

Previous studies using gram-positive and -negative bacteria demonstrated that hydrolysis of silicon nitride (Si3N4) in aqueous suspensions elutes nitrogen and produces gaseous ammonia while buffering pH. According to immunochemistry assays, fluorescence imaging, and in situ Raman spectroscopy, we demonstrate here that the antipathogenic surface chemistry of Si3N4 can be extended to polymethylmethacrylate (PMMA) by compounding it with a minor fraction (~8 vol.%) of Si3N4 particles without any tangible loss in bulk properties. The hydrolytic products, which were eluted from partly exposed Si3N4 particles at the composite surface, exhibited fungicidal action against Candida albicans. Using a specific nitrative stress sensing dye and highly resolved fluorescence micrographs, we observed in situ congestion of peroxynitrite (ONOO−) radicals in the mitochondria of the Candida cells exposed to the PMMA/Si3N4 composite, while these radicals were absent in the mitochondria of identical cells exposed to monolithic PMMA. These in situ observations suggest that the surface chemistry of Si3N4 mimics the antifungal activity of macrophages, which concurrently produce NO radicals and superoxide anions (O2•−) resulting in the formation of candidacidal ONOO−. The fungicidal properties of PMMA/Si3N4 composites could be used in dental appliances to inhibit the uncontrolled growth of Candida albicans and ensuing candidiasis while being synergic with chemoprophylaxis. Statement of significance In a follow-up of previous studies of gram-positive and gram-negative bacteria, we demonstrate here that the antipathogenic surface chemistry of Si3N4 could be extended to polymethylmethacrylate (PMMA) containing a minor fraction (~8 vol.%) of Si3N4 particles without tangible loss in bulk properties. Hydrolytic products eluted from Si3N4 particles at the composite surface exhibited fungicidal action against Candida albicans. Highly resolved fluorescence microscopy revealed congestion of peroxynitrite (ONOO−) radicals in the mitochondria of the Candida cells exposed to the PMMA/Si3N4 composite, while radicals were absent in the mitochondria of identical cells exposed to monolithic PMMA. The fungicidal properties of PMMA/Si3N4 composites could be used in dental appliances to inhibit uncontrolled growth of Candida albicans and ensuing candidiasis in synergy with chemoprophylaxis.

The ability of the new coronavirus SARS‐CoV‐2 to spread and contaminate is one of the determinants of the COVID‐19 pandemic status. SARS‐CoV‐2 has been detected in saliva consistently, with similar sensitivity to that observed in nasopharyngeal swabs. We conducted ultrasound‐guided postmortem biopsies in COVID‐19 fatal cases. Samples of salivary glands (SGs; parotid, submandibular, and minor) were obtained. We analyzed samples using RT‐qPCR, immunohistochemistry, electron microscopy, and histopathological analysis to identify SARS‐CoV‐2 and elucidate qualitative and quantitative viral profiles in salivary glands. The study included 13 female and 11 male patients, with a mean age of 53.12 years (range 8–83 years). RT‐qPCR for SARS‐CoV‐2 was positive in 30 SG samples from 18 patients (60% of total SG samples and 75% of all cases). Ultrastructural analyses showed spherical 70–100 nm viral particles, consistent in size and shape with the Coronaviridae family, in the ductal lining cell cytoplasm, acinar cells, and ductal lumen of SGs. There was also degeneration of organelles in infected cells and the presence of a cluster of nucleocapsids, which suggests viral replication in SG cells. Qualitative histopathological analysis showed morphologic alterations in the duct lining epithelium characterized by cytoplasmic and nuclear vacuolization, as well as nuclear pleomorphism. Acinar cells showed degenerative changes of the zymogen granules and enlarged nuclei. Ductal epithelium and serous acinar cells showed intense expression of ACE2 and TMPRSS receptors. An anti‐SARS‐CoV‐2 antibody was positive in 8 (53%) of the 15 tested cases in duct lining epithelial cells and acinar cells of major SGs. Only two minor salivary glands were positive for SARS‐CoV‐2 by immunohistochemistry. Salivary glands are a reservoir for SARS‐CoV‐2 and provide a pathophysiological background for studies that indicate the use of saliva as a diagnostic method for COVID‐19 and highlight this biological fluid's role in spreading the disease. © 2021 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Coevolutionary adaptation between humans and helminths has developed a finely tuned balance between host immunity and chronic parasitism due to immunoregulation. Given that these reciprocal forces drive selection, experimental models of helminth infection are ideally suited for discovering how host protective immune responses adapt to the unique tissue niches inhabited by these large metazoan parasites. This review highlights the key discoveries in the immunology of helminth infection made over the last decade, from innate lymphoid cells to the emerging importance of neuroimmune connections. A particular emphasis is placed on the emerging areas within helminth immunology where the most growth is possible, including the advent of genetic manipulation of parasites to study immunology and the use of engineered T cells for therapeutic options. Lastly,we cover the status of human challenge trials with helminths as treatment for autoimmune disease, which taken together, stand to keep the study of parasitic worms at the forefront of immunology for years to come.

Arthrospira platensis is a source of proteins, polysaccharides, polyunsaturated fatty acids (in tiny amounts) and phenolic components that give it different biological properties. The type of nutri...

Tributyltin-based (TBT) antifouling paints, widely used for the treatment of flooded surfaces, have been banned in 2008 for their high environmental persistence and bioaccumulation in aquatic organisms. Although it is still present in aquatic ecosystems, oxidative stress driven by TBT has been still poorly investigated in fish. The aim of the study was to examine the time-course stress responses in liver of rainbow trout that received a single intraperitoneal injection of tributyltin chloride (TBTC) or tributyltin ethoxide (TBTE), both at a dose of 0.05 and 0.5 mg/kg. Levels of metallothioneins, total glutathione, malondialdehyde, superoxide dismutase, catalase, glutathione peroxidase and glutathione S-transferase were evaluated at 3 and 6 days post-injection. Tin load was measured in the muscle of the same fish. Differences were observed in the time-course accumulation of tin with a clear dose-response relationship. Although individual oxidative stress biomarkers varied, the biomarker profile indicated different stress mechanisms caused by both TBTC and TBTE. The weak induction of metal-trapping metallothioneins and the changes of oxidative stress biomarkers suggested a stress-pressure in both TBT-treated trout, advising for an ecotoxicological risk for freshwater ecosystems.

Objective: The study was undertaken to identify a monogenic cause of early onset, generalized dystonia. Methods: Methods consisted of genome-wide linkage analysis, exome and Sanger sequencing, clinical neurological examination, brain magnetic resonance imaging, and protein expression studies in skin fibroblasts from patients. Results: We identified a heterozygous variant, c.388G>A, p.Gly130Arg, in the eukaryotic translation initiation factor 2 alpha kinase 2 (EIF2AK2) gene, segregating with early onset isolated generalized dystonia in 5 patients of a Taiwanese family. EIF2AK2 sequencing in 191 unrelated patients with unexplained dystonia yielded 2 unrelated Caucasian patients with an identical heterozygous c.388G>A, p.Gly130Arg variant, occurring de novo in one case, another patient carrying a different heterozygous variant, c.413G>C, p.Gly138Ala, and one last patient, born from consanguineous parents, carrying a third, homozygous variant c.95A>C, p.Asn32Thr. These 3 missense variants are absent from gnomAD, and are located in functional domains of the encoded protein. In 3 patients, additional neurological manifestations were present, including intellectual disability and spasticity. EIF2AK2 encodes a kinase (protein kinase R [PKR]) that phosphorylates eukaryotic translation initiation factor 2 alpha (eIF2α), which orchestrates the cellular stress response. Our expression studies showed abnormally enhanced activation of the cellular stress response, monitored by PKR-mediated phosphorylation of eIF2α, in fibroblasts from patients with EIF2AK2 variants. Intriguingly, PKR can also be regulated by PRKRA (protein interferon-inducible double-stranded RNA-dependent protein kinase activator A), the product of another gene causing monogenic dystonia. Interpretation: We identified EIF2AK2 variants implicated in early onset generalized dystonia, which can be dominantly or recessively inherited, or occur de novo. Our findings provide direct evidence for a key role of a dysfunctional eIF2α pathway in the pathogenesis of dystonia. ANN NEUROL 2021;89:485–497.

The nature of endometrial morular metaplasia (MorM) is still unknown. The nuclear β-catenin accumulation and the not rare ghost cell keratinization suggest a similarity with hard keratin-producing odontogenic and hair matrix tumors rather than with squamous differentiation. We aimed to compare MorM to hard keratin-producing tumors. Forty-one hard keratin-producing tumors, including 26 hair matrix tumors (20 pilomatrixomas and 6 pilomatrix carcinomas) and 15 odontogenic tumors (adamantinomatous craniopharyngiomas), were compared to 15 endometrioid carcinomas with MorM with or without squamous/keratinizing features. Immunohistochemistry for β-catenin, CD10, CDX2, ki67, p63, CK5/6, CK7, CK8/18, CK19, and pan-hard keratin was performed; 10 cases of endometrioid carcinomas with conventional squamous differentiation were used as controls. In adamantinomatous craniopharyngiomas, the β-catenin-accumulating cell clusters (whorl-like structures) were morphologically similar to MorM (round syncytial aggregates of bland cells with round-to-spindled nuclei and profuse cytoplasm), with overlapping squamous/keratinizing features (clear cells with prominent membrane, rounded squamous formations, ghost cells). Both MorM and whorl-like structures consistently showed positivity for CD10 and CDX2, with low ki67; cytokeratins pattern was also overlapping, although more variable. Hard keratin was focally/multifocally positive in 8 MorM cases and focally in one conventional squamous differentiation case. Hair matrix tumors showed no morphological or immunophenotypical overlap with MorM. MorM shows wide morphological and immunophenotypical overlap with the whorl-like structures of adamantinomatous craniopharyngiomas, which are analogous to enamel knots of tooth development. This suggests that MorM might be an aberrant mimic of odontogenic differentiation. Supplementary Information The online version contains supplementary material available at 10.1007/s00428-021-03060-2.

Feline coronavirus (FCoV) exists as two different genotypes, FCoV type I and II, each including two biotypes, feline enteric coronavirus (FECV) and feline infectious peritonitis virus (FIPV), the latter being a virulent variant originating from the former virus. Recently, two amino acid substitutions, M1058L and S1060A, within the spike protein have been associated to the FECV/FIPV virulence change. In this study, we have analysed the frequency of detection of such mutations in FIPV and FECV strains circulating in Italian cats and obtained information about their evolutionary relationships with reference isolates. A total of 40 FCoV strains, including 19 strains from effusions or tissue samples of FIP cats and 21 strains from faecal samples of non-FIP cats, were analysed. Mutation M1058L was detected in 16/18 FCoV-I and 1/1 FCoV-II strains associated with FIP, while change S1060A was presented by two FIPV strains. By phylogenetic analysis, FCoV sequences clustered according to the genotype but not according to the biotype, with FECV/FIPV strains recovered from the same animal being closely related. Further studies are needed to better define the genetic signatures associated with the FECV/FIPV virulence shift.

Monitoring the invasion process of the Asian tiger mosquito Aedes albopictus and its interaction with the contender Aedes aegypti, is critical to prevent and control the arthropod-borne viruses (i.e., Arboviruses) they transmit to humans. Generally, the superior ecological competitor Ae. albopictus displaces Ae. aegypti from most geographic areas, with the combining factors of biology and environment influencing the competitive outcome. Nonetheless, detailed studies asserting displacement come largely from sub-tropical areas, with relatively less effort being made in tropical environments, including no comprehensive research about Aedes biological interactions in Mesoamerica. Here, we examine contemporary and historical mosquito surveillance data to assess the role of shifting abiotic conditions in shaping the spatiotemporal distribution of competing Aedes species in the Republic of Panama. In accordance with prior studies, we show that Ae. albopictus has displaced Ae. aegypti under suboptimal wet tropical climate conditions and more vegetated environments within the southwestern Azuero Peninsula. Conversely, in the eastern Azuero Peninsula, Ae. aegypti persists with Ae. albopictus under optimal niche conditions in a dry and more seasonal tropical climate. While species displacement was stable over the course of two years, the presence of both species generally appears to fluctuate in tandem in areas of coexistence. Aedes albopictus was always more frequently found and abundant regardless of location and climatic season. The heterogenous environmental conditions of Panama shape the competitive outcome and micro-geographic distribution of Aedes mosquitoes, with potential consequences for the transmission dynamics of urban and sylvatic zoonotic diseases.

The demand for natural antioxidants to be used in food industry is increasing, as synthetic antioxidants are toxic and have high production costs. Specifically, food processing and preservation require antioxidants resistant to thermal sterilization processes. In this study, twenty-five strains among microalgae and cyanobacteria were screened as antioxidants producers. The species Enallax sp., Synechococcus bigranulatus and Galdieria sulphuraria showed the highest content of chlorophyll a and total carotenoids. In vitro stability and antioxidant activity of the ethanolic extracts were performed. The results revealed that pigments present in the extracts, obtained from the previously mentioned species, were stable at room temperature and exhibited in vitro free radical scavenging potential with IC50 values of 0.099 ± 0.001, 0.048 ± 0.001 and 0.13 ± 0.02 mg mL-1, respectively. Biocompatibility assay showed that the extracts were not toxic on immortalized cell lines. The antioxidant activity was also tested on a cell-based model by measuring intracellular ROS levels after sodium arsenite treatment. Noteworthy, extracts were able to exert the same protective effect, before and after the pasteurization process. Results clearly indicate the feasibility of obtaining biologically active and thermostable antioxidants from microalgae. Green solvents can be used to obtain thermo-resistant antioxidants from cyanobacteria and microalgae which can be used in the food industry. Thus, the substitution of synthetic pigments with natural ones is now practicable. Graphical abstract: [Figure not available: see fulltext.].

Converging clinical and preclinical evidence demonstrates that depressive phenotypes are associated with synaptic dysfunction and dendritic simplification in cortico-limbic glutamatergic areas. On the other hand, the rapid antidepressant effect of acute ketamine is consistently reported to occur together with the rescue of dendritic atrophy and reduction of spine number induced by chronic stress in the hippocampus and prefrontal cortex of animal models of depression. Nevertheless, the molecular mechanisms underlying these morphological alterations remain largely unknown. Here, we found that miR-9-5p levels were selectively reduced in the hippocampus of rats vulnerable to Chronic Mild Stress (CMS), while acute subanesthetic ketamine restored its levels to basal condition in just 24h; miR-9-5p expression inversely correlated with the anhedonic phenotype. A decrease of miR-9-5p was reproduced in an in vitro model of stress, based on primary hippocampal neurons incubated with the stress hormone corticosterone. In both CMS animals and primary neurons, decreased miR-9-5p levels were associated with dendritic simplification, while treatment with ketamine completely rescued the changes. In vitro modulation of miR-9-5p expression showed a direct role of miR-9-5p in regulating dendritic length and spine density in mature primary hippocampal neurons. Among the putative target genes tested, Rest and Sirt1 were validated as biological targets in primary neuronal cultures. Moreover, in line with miR-9-5p changes, REST protein expression levels were remarkably increased in both CMS vulnerable animals and corticosterone-treated neurons, while ketamine completely abolished this alteration. Finally, the shortening of dendritic length in corticosterone-treated neurons was shown to be partly rescued by miR-9-5p overexpression and dependent on REST protein expression. Overall, our data unveiled the functional role of miR-9-5p in the remodeling of dendritic arbor induced by stress/corticosterone in vulnerable animals and its rescue by acute antidepressant treatment with ketamine., Highlights • miR-9-5p is reduced in the hippocampus of stress-vulnerable rats. • miR-9-5p is reduced in primary neurons treated with corticosterone. • Acute ketamine restores stress-induced morphological changes and miR-9-5p levels. • miR-9-5p directly modulates dendritic length and spine number in primary neurons. • REST is a validated target of miR-9-5p.

The systematic status of the controversial crustacean family Leptobathynellidae is investigated using molecular and morphological methods in this study. Partial sequences of the nuclear 18S gene are studied from 28 genera of Bathynellacea from several continents. The analysis includes some of the most plesiomorphic genera of the family Parabathynellidae, such as Billibathynella and Brevisomabathynella from Australia; Habrobathynella and Parvulobathynella from India; the diverse Iberobathynella; the cosmopolitan genus Hexabathynella; and representative genera of two subfamilies of Bathynellidae (Gallobathynellinae and Bathynellinae). We used a molecular approach to analyse the systematic relationships amongst 64 species from Europe, North America, Australia and Asia, and review the morphological characters relevant at the family level. The molecular phylogeny clearly shows the presence of three highly divergent clades that could represent the three families. This is the first molecular phylogenetic reconstruction of Bathynellacea that can be used to: (1) verify the validity of Leptobathynellidae, (2) explore the diversity of the families and (3) explore the phylogenetic relationships among families. We propose a plausible evolutionary scenario for the order Bathynellacea.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the causal agent of COVID-19 and stands at the center of the current global human pandemic, with death toll exceeding one million. The urgent need for a vaccine has led to the development of various immunization approaches. mRNA vaccines represent a cell-free, simple, and rapid platform for immunization, and therefore have been employed in recent studies toward the development of a SARS-CoV-2 vaccine. Herein, we present the design of an mRNA vaccine, based on lipid nanoparticles (LNPs)-encapsulated SARS-CoV-2 human Fc-conjugated receptor-binding domain (RBD-hFc). Several ionizable lipids have been evaluated in vivo in a luciferase (luc) mRNA reporter assay, and two leading LNPs formulations have been chosen for the subsequent RBD-hFc mRNA vaccine strategy. Intramuscular administration of LNP RBD-hFc mRNA elicited robust humoral response, a high level of neutralizing antibodies and a Th1-biased cellular response in BALB/c mice. The data in the current study demonstrate the potential of these lipids as promising candidates for LNP-based mRNA vaccines in general and for a COVID19 vaccine in particular.

Metabolic transformation is a hallmark of cancer and a critical target for cancer therapy. Cancer metabolism and behaviour are regulated by cell-intrinsic factors as well as metabolite availability in the tumour microenvironment (TME). This metabolic niche within the TME is shaped by four tiers of regulation: (1) intrinsic tumour cell metabolism, (2) interactions between cancer cells and non-cancerous cells, (3) tumour location and heterogeneity and (4) whole-body metabolic homeostasis. Here, we define these modes of metabolic regulation and review how distinct cell types contribute to the metabolite composition of the TME. Finally, we connect these insights to understand how each of these tiers offers unique therapeutic potential to modulate the metabolic profile and function of all cells inhabiting the TME. The tumour microenvironment (TME) is a unique cellular and metabolic landscape. Elia and Haigis describe how metabolism influences, and is affected by, the complexity of cellular interactions within the TME and highlight opportunities for therapeutic intervention.

Sacubitril/valsartan (S/V) is a new drug which has been recently recommended by the international guidelines for the treatment of patients with chronic heart failure (HF) with reduced ejection fraction (HFrEF). Compared to angiotensin-converting enzyme inhibitors (ACEi) and angiotensin II receptor blockers (ARBs), S/V is associated with a better cardiovascular outcome, and to a greater beneficial effect on myocardial reverse remodeling. Recent evidence has shown that S/V is not only recommended in chronic patients but it is also approved in the acute setting; moreover, its safety and tolerability have been demonstrated also in the pediatric population. This review summarizes data on the effectiveness and tolerability of S/V in HFrEF patients and offers practical insights to manage this drug in every setting, providing an overview from randomized clinical trials' data to real-world evidence.

This bibliometric study investigates the research methodology practices of Master of Arts in Information Studies (MAIS) students at the University of Dar es Salaam, Tanzania. The study established students’ insufficient understanding and application of research methodology concepts. Survey research was predominant, with purposive and convenience non-probability sampling methods being extensively used. Simple random sampling and stratified sampling were the probability sampling methods highly used. Findings further show advanced qualitative and quantitative data analyses were inadequately applied. In practice, the study findings can help Library and Information Science institutions around the globe improve teaching research methodology to produce quality theses with logical conclusions which can develop new theories. Quality theses can translate into increased quantity and quality of journal articles and growth of the Library and Information Science discipline. Thus, there is a need to strengthen research methodology training for students and lecturers to generate generalizable findings that meet diverse needs.