Applying two approaches to detect unmeasured confounding due to time-varying variables in a self-controlled risk interval design evaluating COVID-19 vaccine safety signals, using myocarditis as a case example.

We test the robustness of the self-controlled risk interval (SCRI) design in a setting where time between doses may introduce time-varying confounding, using both negative control outcomes (NCOs) and quantitative bias analysis (QBA). All vaccinated cases identified from 5 European databases between 1 September 2020 and end of data availability were included. Exposures were doses 1-3 of the Pfizer, Moderna, AstraZeneca, and Janssen COVID-19 vaccines; outcomes were myocarditis and otitis externa (NCO). The SCRI used a 60-day control window and dose-specific 28-day risk windows, stratified by vaccine brand and adjusted for calendar time. The QBA included two scenarios: (i) baseline probability of the confounder was higher in the control window and (ii) vice versa. The NCO was not associated with any of the COVID-19 vaccine types or doses except Moderna dose 1 (IRR = 1.09, 95%CI 1.01-1.09). The QBA suggested even the strongest literature-reported confounder (COVID-19; RRmyocarditis = 18.3) could only explain away part of the observed effect from IRR = 3 to IRR = 1.40. The SCRI seems robust to unmeasured confounding in the COVID-19 setting, although a strong unmeasured confounder could bias the observed effect upward. Replication of our findings for other safety signals would strengthen this conclusion.

Describing diversity of real world data sources in pharmacoepidemiologic studies: The DIVERSE scoping review.

PURPOSE: Real-world evidence (RWE) is increasingly used for medical regulatory decisions, yet concerns persist regarding its reproducibility and hence validity. This study addresses reproducibility challenges associated with diversity across real-world data sources (RWDS) repurposed for secondary use in pharmacoepidemiologic studies. Our aims were to identify, describe and characterize practices, recommendations and tools for collecting and reporting diversity across RWDSs, and explore how leveraging diversity could improve the quality of evidence. METHODS: In a preliminary phase, keywords for a literature search and selection tool were designed using a set of documents considered to be key by the coauthors. Next, a systematic search was conducted up to December 2021. The resulting documents were screened based on titles and abstracts, then based on full texts using the selection tool. Selected documents were reviewed to extract information on topics related to collecting and reporting RWDS diversity. A content analysis of the topics identified explicit and latent themes. RESULTS: Across the 91 selected documents, 12 topics were identified: 9 dimensions used to describe RWDS (organization accessing the data source, data originator, prompt, inclusion of population, content, data dictionary, time span, healthcare system and culture, and data quality), tools to summarize such dimensions, challenges, and opportunities arising from diversity. Thirty-six themes were identified within the dimensions. Opportunities arising from data diversity included multiple imputation and standardization. CONCLUSIONS: The dimensions identified across a large number of publications lay the foundation for formal guidance on reporting diversity of data sources to facilitate interpretation and enhance replicability and validity of RWE.

APOL1 Gene Variants and Risk for Cardiovascular Disease.

INTRODUCTION: The association of APOL1 risk variants with cardiovascular risk and cardiovascular disease (CVD) in observational and clinical trials has had inconsistent results. We aim to assess the relationship between the presence of APOL1 risk variants and the CVD risk in Afro-descendant patients with end-stage renal disease (ESRD). METHODS: We performed an observational, cross-sectional study of Afro-descendant adult patients with ESRD who were on the waitlist for a kidney transplant. Associations of APOL1 genotypes (high-risk [HR] = 2 alleles; low-risk [LR] = 0 or 1 allele) with cardiovascular risk were the primary clinical endpoint. The relation was evaluated using univariate and multivariate analysis. RESULTS: We enrolled a total of 102 patients with ESRD; 37% (38 patients) had APOL1 HR status with two alleles in homozygous (G1/G1 = 21 and G2/G2 = 3) or compound heterozygote (G1/G2 = 14) form and 63% (64 patients) had APOL1 LR status. No significant association was found between HR APOL1 genotypes and high cardiovascular risk (in adjusted Colombia Framingham Risk Score). APOL1 HR versus LR variants were not independently associated with left ventricular hypertrophy or systolic dysfunction. No cardiovascular deaths occurred during the follow-up. CONCLUSION: In Afro-descendent patients with ESRD, APOL1 HR status is not associated with the increase in cardiovascular risk profile and metabolic disturbances.

Effects of the COVID-19 pandemic on food insecurity in El Salvador during 2020.

Objective: This study sought to quantify the prevalence of food insecurity among Salvadorian households, to identify the determinants of food insecurity and to explore the impact of the COVID-19 pandemic on food insecurity. Methods: A nationwide, representative random sample of 2358 households was used for this cross-sectional study. The Household Hunger Scale (HHS) was used to assess the prevalence of food insecurity during a 30-day period. For comparison, three items were used from the Household Food Insecurity Experience Scale (HFIES), which measures hunger occurring during a 12-month time frame. For determinant analysis, binary logistic regression was used for the HHS and ordered logistic regression for the HFIES. Results: The prevalence of food insecurity was 6.45% (152/2356) among Salvadorian households when the HHS was used, affecting 5.48% (129/2356) to a moderate degree and 0.98% (23/2356) to a severe degree. The prevalence significantly increased when the HFIES scale items were used, with 35.41% (835/2358) of households being affected, a figure closer to the national poverty level. Determinants of food insecurity according to the HHS included agricultural problems (P = 0.00, odds ratio [OR] =1.69), the household's prepandemic income (P = 0.00, OR = 0.48) and higher educational levels (i.e. having a secondary education [P = 0.00, OR = 0.31], technical [P = 0.03, OR = 0.24] or university education [P = 0.00, OR = 0.05]). When using the HFIES, the determinants were similar (i.e. income, agricultural problems, educational level). In more than 94% (744/785) of households, participants reported that food insecurity was exacerbated by the COVID-19 pandemic. Conclusions: When compared with other relevant international studies, the prevalence of food insecurity identified using the HHS - only 6.45% - was low for El Salvador. However, when using the HFIES scale, the prevalence rose to 35.41% of households. Some determinants align with previous studies, namely income, educational level and agricultural problems. The COVID-19 pandemic appeared to have direct effects on food insecurity.

The Emergence of New Catalytic Abilities in an Endoxylanase from Family GH10 by Removing an Intrinsically Disordered Region.

Endoxylanases belonging to family 10 of the glycoside hydrolases (GH10) are versatile in the use of different substrates. Thus, an understanding of the molecular mechanisms underlying substrate specificities could be very useful in the engineering of GH10 endoxylanases for biotechnological purposes. Herein, we analyzed XynA, an endoxylanase that contains a (ß/α)8-barrel domain and an intrinsically disordered region (IDR) of 29 amino acids at its amino end. Enzyme activity assays revealed that the elimination of the IDR resulted in a mutant enzyme (XynAΔ29) in which two new activities emerged: the ability to release xylose from xylan, and the ability to hydrolyze p-nitrophenyl-ß-d-xylopyranoside (pNPXyl), a substrate that wild-type enzyme cannot hydrolyze. Circular dichroism and tryptophan fluorescence quenching by acrylamide showed changes in secondary structure and increased flexibility of XynAΔ29. Molecular dynamics simulations revealed that the emergence of the pNPXyl-hydrolyzing activity correlated with a dynamic behavior not previously observed in GH10 endoxylanases: a hinge-bending motion of two symmetric regions within the (ß/α)8-barrel domain, whose hinge point is the active cleft. The hinge-bending motion is more intense in XynAΔ29 than in XynA and promotes the formation of a wider active site that allows the accommodation and hydrolysis of pNPXyl. Our results open new avenues for the study of the relationship between IDRs, dynamics and activity of endoxylanases, and other enzymes containing (ß/α)8-barrel domain.

Midpalatal suture density ratio after rapid maxillary expansion evaluated by cone-beam computed tomography.

INTRODUCTION: Identifying the stage of midpalatal suture (MPS) fusion is essential for rapid maxillary expansion (RME). The objective of this study was to evaluate the relationship between MPS density ratio and the skeletal response after RME using cone-beam computed tomography (CBCT). METHODS: CBCT scans of 64 patients (age, 13.81 ± 1.68 years; 42 females and 22 males) who underwent RME as part of comprehensive orthodontic treatment were used for this study. Two CBCT records were made at T1 (pre-RME) and T2 (post-RME). MPS density was assessed by gray scale and the skeletal effects through linear measurements: greater palatine foramina, the width of the nasal cavity (NW), infraorbital foramina. The data were collected at 4 different times: baseline, 24 hours, 48 hours, and 4 weeks. RESULTS: There was no statistically significant correlation between MPS density ratio and greater palatine foramina or infraorbital foramina. Negative correlations with MPS density ratio are presented with the difference of age post-RME and pre-RME (r = -0.46, P = 0.033) in males, and NW (r = -0.41, P = 0.007) in females. A significant association between gender and the MPS density ratio was found (P = 0.015) in females. CONCLUSIONS: There is a weak negative correlation between the MPS density ratio and the difference of age post-RME and pre-RME in males and the NW in females. Females presented greater density ratios than males. MPS density ratio through CBCT does not appear to be an adequate parameter to predict the skeletal effect in adolescent patients and young adults.

Preprints in times of COVID19: the time is ripe for agreeing on terminology and good practices.

Over recent years, the research community has been increasingly using preprint servers to share manuscripts that are not yet peer-reviewed. Even if it enables quick dissemination of research findings, this practice raises several challenges in publication ethics and integrity. In particular, preprints have become an important source of information for stakeholders interested in COVID19 research developments, including traditional media, social media, and policy makers. Despite caveats about their nature, many users can still confuse pre-prints with peer-reviewed manuscripts. If unconfirmed but already widely shared first-draft results later prove wrong or misinterpreted, it can be very difficult to "unlearn" what we thought was true. Complexity further increases if unconfirmed findings have been used to inform guidelines. To help achieve a balance between early access to research findings and its negative consequences, we formulated five recommendations: (a) consensus should be sought on a term clearer than 'pre-print', such as 'Unrefereed manuscript', "Manuscript awaiting peer review" or ''Non-reviewed manuscript"; (b) Caveats about unrefereed manuscripts should be prominent on their first page, and each page should include a red watermark stating 'Caution-Not Peer Reviewed'; (c) pre-print authors should certify that their manuscript will be submitted to a peer-review journal, and should regularly update the manuscript status; (d) high level consultations should be convened, to formulate clear principles and policies for the publication and dissemination of non-peer reviewed research results; (e) in the longer term, an international initiative to certify servers that comply with good practices could be envisaged.

Regulatory reliance to approve new medicinal products in Latin American and Caribbean countries.

OBJECTIVE: To describe the current status of regulatory reliance in Latin America and the Caribbean (LAC) by assessing the countries' regulatory frameworks to approve new medicines, and to ascertain, for each country, which foreign regulators are considered as trusted regulatory authorities to rely on. METHODS: Websites from LAC regulators were searched to identify the official regulations to approve new drugs. Data collection was carried out in December 2019 and completed in June 2020 for the Caribbean countries. Two independent teams collected information regarding direct recognition or abbreviated processes to approve new drugs and the reference (trusted) regulators defined as such by the corresponding national legislation. RESULTS: Regulatory documents regarding marketing authorization were found in 20 LAC regulators' websites, covering 34 countries. Seven countries do not accept reliance on foreign regulators. Thirteen regulatory authorities (Argentina, Colombia, Costa Rica, Dominican Republic, Ecuador, El Salvador, Guatemala, Mexico, Panama, Paraguay, Peru, Uruguay, and the unique Caribbean Regulatory System for 15 Caribbean States) explicitly accept relying on marketing authorizations issued by the European Medicines Agency, United States Food and Drug Administration, and Health Canada. Ten countries rely also on marketing authorizations from Australia, Japan, and Switzerland. Argentina, Brazil, Chile, and Mexico are reference authorities for eight LAC regulators. CONCLUSIONS: Regulatory reliance has become a common practice in the LAC region. Thirteen out of 20 regulators directly recognize or abbreviate the marketing authorization process in case of earlier approval by a regulator from another jurisdiction. The regulators most relied upon are the European Medicines Agency, United States Food and Drug Administration, and Health Canada.

Genomic Epidemiology of Severe Acute Respiratory Syndrome Coronavirus 2, Colombia.

Coronavirus disease (COVID-19) in Colombia was first diagnosed in a traveler arriving from Italy on February 26, 2020. However, limited data are available on the origins and number of introductions of COVID-19 into the country. We sequenced the causative agent of COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), from 43 clinical samples we collected, along with another 79 genome sequences available from Colombia. We investigated the emergence and importation routes for SARS-CoV-2 into Colombia by using epidemiologic, historical air travel, and phylogenetic observations. Our study provides evidence of multiple introductions, mostly from Europe, and documents >12 lineages. Phylogenetic findings validate the lineage diversity, support multiple importation events, and demonstrate the evolutionary relationship of epidemiologically linked transmission chains. Our results reconstruct the early evolutionary history of SARS-CoV-2 in Colombia and highlight the advantages of genome sequencing to complement COVID-19 outbreak investigations.

Silver nanoparticles from Justicia spicigera and their antimicrobial potentialities in the biocontrol of foodborne bacteria and phytopathogenic fungi.

In the present work, the biosynthesis of silver-nanoparticles (AgNP) was evaluated using the aqueous extract from Justicia spicigera. The obtained silver nanoparticles were characterized using UV-visible spectroscopic techniques, energy dispersive X-ray spectrometers (EDS), zeta potential and dynamic light scattering. The antimicrobial activity of biosynthesized AgNP was tested against foodborne bacteria (Bacillus cereus, Klebsiella pneumoniae and Enterobacter aerogenes) and phytopathogenic fungi (Colletotrichum sp., Fusarium solani, Alternaria alternata and Macrophomina phaseolina). The elemental profile of synthesized nanoparticles using J. spicigera shows higher counts at 3keV due to silver, confirming the formation of silver nanoparticles. Scanning electron microscopy (SEM) analysis showed a particle size between 86 and 100nm with spherical morphology. AgNP showed effective antibacterial and antifungal activity against the tested organisms principally with B. cereus, K. pneumoniae, E. aerogenes, A. alternata and M. phaseolina. Therefore, further studies are needed to confirm the potential of AgNP from J. spicigera in the control of indicator organisms under field conditions.

Cold-active pectinolytic activity produced by filamentous fungi associated with Antarctic marine sponges.

BACKGROUND: Pectinase enzymes catalyze the breakdown of pectin, a key component of the plant cell wall. At industrial level, pectinases are used in diverse applications, especially in food-processing industry. Currently, most of the industrial pectinases have optimal activity at mesophilic temperatures. On the contrary, very little is known about the pectinolytic activities from organisms from cold climates such as Antarctica. In this work, 27 filamentous fungi isolated from marine sponges collected in King George Island, Antarctica, were screened as new source of cold-active pectinases. RESULTS: In semi-quantitative plate assays, 8 out 27 of these isolates showed pectinolytic activities at 15 °C and one of them, Geomyces sp. strain F09-T3-2, showed the highest production of pectinases in liquid medium containing pectin as sole carbon source. More interesting, Geomyces sp. F09-T3-2 showed optimal pectinolytic activity at 30 °C, 10 °C under the temperature of currently available commercial mesophilic pectinases. CONCLUSION: Filamentous fungi associated with Antarctic marine sponges are a promising source of pectinolytic activity. In particular, pectinases from Geomyces sp. F09-T3-2 may be potentially suitable for biotechnological applications needing cold-active pectinases. To the best of our knowledge, this is the first report describing the production of pectinolytic activity from filamentous fungi from any environment in Antarctica.

Changes in the phenylalanine ammonia lyase activity, total phenolic compounds, and flavonoids in Prosopis glandulosa treated with cadmium and copper.

The aim of the present work is to evaluate the changes on the phenylalanine ammonia lyase (PAL) activity, phenolic compounds accumulation and photochemical efficiency in leaves of P. glandulosa treated with Cd2+ (0.001 M) and Cu2+ (0.52 M) concentrations for 96 h under hydroponic conditions. The results showed that only leaves treated with copper had a decrease in photochemical efficiency and leaf epidermal polyphenols in P. glandulosa leaves after 96 h of exposure. On the other hand the reverse-phase HPLC analysis revealed higher levels of phenolic compound (gallic, vanillic and caffeic acids) and flavonoids (rutin and kaempferol-3-O-glucosides) in plant leaves from Cu and Cd-treatments with respect to control plants. Finally, highest increments in PAL activity was observed in extracts of leaves treated with Cu and Cd (about 205 and 284%), respectively, with respect to control plants after 96 h treatment. These suggest that activation of phenylpropanoid pathway represent a source of nonenzymatic antioxidants that protect at P. glandulosa against oxidative stress when exposed to cadmium and copper. Hence future studies are necessary to elucidate the participation of phenylpropanoid pathway in the reduction of metal toxicity in Prosopis species.

Systematic review of cross-national drug utilization studies in Latin America: methods and comparability.

PURPOSE: We systematically review the cross-national drug utilization studies performed in Latin America (LA) in order to analyze the methods applied and assess the validity of the data to ensure the comparability. METHODS: A systematic search in Medline, Embase, and BIREME was performed. Drug utilization studies including LA countries and comparing drug exposure data on volume were included. The data validity was judged independently by two authors as having low, medium, high, or unclear risk of bias. RESULTS: Out of 1191 articles, 25 were kept for full text reading. Finally, five studies were selected. Eight different Latin American countries were involved in the comparisons. The selected studies analyzed wholesale data from a private research company collecting information from the private healthcare sector. In three studies, a high risk of bias in the extrapolation method applied was identified. In one study, a risk of data collection bias was detected. The most frequent limitation detected by the original authors was related to the unavailability of information from the public sector in LA. CONCLUSION: Drug utilization studies comparing data cross-nationally are scarce in LA. In general, validity of the comparisons is hampered by a potential risk of extrapolation bias given the lack of available data on drug consumption from the public healthcare sector. Setting up systems to remediate this situation is a future challenge for researchers and (supra)national authorities in the region.

[Hyperuricaemia and metabolic syndrome in obese children and adolescents]. / Hiperuricemia y componentes del síndrome metabólico en niños y adolescentes obesos.

INTRODUCTION: Hyperuricaemia has been suggested as an additional metabolic factor in adult obese patients, but it has not been sufficiently studied in paediatric. OBJECTIVES: To assess the relationship between serum uric acid levels (SUAL) with the level of general and visceral obesity, and other biochemical parameters in children and adolescents of Santiago, Chile. SUBJECTS AND METHOD: A cross sectional study was conducted on 770 children and adolescents (ages: 6-15 y.) from a public school in Santiago, Chile, of whom 227 (29%) were obese (BMI>2 SD, WHO growth standards). Ninety subjects were randomly selected and 77 with no other chronic disease (41 males) accepted to participate. Data was collected on weight, stature, abdominal circumference (AC), visceral adiposity using ultrasound, and other biochemical measurements including fasting glucose, insulin, serum lipids, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and SUAL. RESULTS: The mean SUAL was 0.200±0.065 mmol/L, and was increased in children with hyperinsulinism (adjusted by age: 0.221±0.075 vs. 0.183±0.054 mmol/L; P<.01), with no significant differences according to HOMA. Differences were also found between children with ALT>or<26 U/mL: 0.238±0.070 vs. 0.178±0.054 mmol/L, P<.001. The logistic regression showed the increased SUAL was only associated with increased ALT. No significant differences were found in general or visceral adiposity measurements or fatty liver. CONCLUSIONS: Children and adolescents from Santiago, Chile have higher uric acid serum uric acid levels as well as an association with increased ALT and insulin. It is demonstrated in this study that uric acid should be measured in obese children and adolescents, and in their follow up.

[Antifungal effect of phenolic and carotenoids extracts from chiltepin (Capsicum annum var. glabriusculum) on Alternaria alternata and Fusarium oxysporum]. / Efecto antifúngico de extractos fenólicos y de carotenoides de chiltepín (Capsicum annum var. glabriusculum) en Alternaria alternata y Fusarium oxysporum.

The effect of phenolic and carotenoid extracts from chiltepin fruits on mycelial growth and the inhibition of conidial germination of Alternaria alternata and Fusarium oxysporum were investigated in the present work. Phenolic extracts inhibited mycelial growth of A.alternata by 38.46%, and significantly reduced conidial germination on the fifth day after treatment to 92% in relation to control. No significant changes were observed in the inhibition of mycelial growth in Fusarium oxysporum; however, the number of germinated conidia was reduced, showing 85% inhibition five days after treatment in relation to control. Moreover, carotenoid extracts showed 38.5% inhibition of mycelial growth and 85.3% inhibition of conidial germination of A.alternata, five days after treatment. Carotenoid extracts showed less inhibition of mycelial growth (20.3%) in F.oxysporum, with respect to A.alternata; while there was greater inhibition of conidial germination (96%) on the fifth day after treatment. Phenolic and carotenoid extracts from chiltepin may be a promising alternative as a natural fungicide against fungi of agricultural importance.

[Migrations in Latin American countries. Characteristics of the pediatric population]. / Migraciones en países de América Latina. Características de la población pediátrica.

INTRODUCTION: Migration is a growing phenomenon among Latin American countries (LAC) as well as others; however, scarce information is available studying its impact on paediatric groups and its association with socioeconomic variables. OBJECTIVE: To study the association among socioeconomic variables and the immigration rate of paediatric population in LAC. MATERIAL AND METHODS: Official rates of migration of LAC were obtained from: International Organization for Migration, Pan American Health Organization, and United Nations Development Programme. Demographic and socioeconomic information was also obtained for: gross domestic product (GDP), human development index (HDI), Gini coefficient of inequality (GC), alphabetization rate for adults (AA), net migration rate (NMR), and immigration of children<15 years (IM15). Description, linear correlations and analysis of differences between groups of countries were assessed. RESULTS: The NMR was positive for Costa Rica, Panama, Venezuela, Chile and Argentina. No association among NMR and GDP, HDI, GC, AA was found. A correlation of IM15 was found with: GC (r=0.668, P=.01), with GDP (r=-0.720; P=.01), AA (r=-0.755; P=.01) and with HDI (r=-0.799; P=.01). Rate of IM15 was lower in LA countries with advanced/medium development (GDP>median) vs those with low development (Fisher, P<.0001). CONCLUSIONS: There is a direct inverse association between GDP per capita, HDI, AA and GC and the proportion of each country IN15. We did not observe an association between NMR and HDI, AA, and GC. The health impact of these migrations should be analysed.

[Effect of supplementation with a single dose of vitamin D in children with cerebral palsy. Preliminary randomised controlled study]. / Efecto de la suplementación con vitamina D en monodosis en niños con parálisis cerebral. Estudio preliminar controlado y aleatorizado.

INTRODUCTION: Children with cerebral palsy (CP) have an increased risk of vitamin D (VD) deficiency. Although there are many studies on VD and CP, there is limited information about VD supplementation in these patients. OBJECTIVE: To evaluate the effect of supplementation with a single dose of VD on the plasma concentrations of 25-hydroxy-vitamin-D (25OHD) in children with CP. PATIENTS AND METHOD: Prospective-randomised-controlled-trial, including 30 Chilean children (19 males) with CP, median age 9.9 years (6.2-13.5). Clinical and biochemical variables including 25OHD, were recorded (time 0 and 8 weeks). Patients were allocated to the supplemented (S) group receiving 100,000 IU oral D3 at baseline, and compared with the placebo (P) group. RESULTS: Among clinical features are highlighted: gastrostomy (60%), underweight (30%), bed-ridden (93.3%), antiepileptic drugs (70%), and 43.3% used VD metabolism inducing antiepileptics. Baseline biochemical measurements were normal. The 25OHD was insufficient in 4/30 and deficient in 6/30. 25OHD levels were not associated with the variables studied. Eight patients completed the study in the S group, and 10 in P group. The placebo and supplementation groups had no significant difference in baseline variables. Serum calcium, phosphate, and alkaline phosphatase levels at 8 weeks were normal in both groups, with no statistically significant differences. 25OHD in the P group was normal in 6/10, and insufficient+deficient in 4/10, and the S group was normal in all (8/8) (exact Fisher test P=.07). CONCLUSIONS: A single dose of 100,000 IU VD could normalise the concentrations of 25OHD after 8 weeks of supplementation in Children with CP, but more studies are required to confirm these results.

Sugar transporters spatially organize microbiota colonization along the longitudinal root axis of Arabidopsis.

Plant roots are functionally heterogeneous in cellular architecture, transcriptome profile, metabolic state, and microbial immunity. We hypothesized that axial differentiation may also impact spatial colonization by root microbiota along the root axis. We developed two growth systems, ArtSoil and CD-Rhizotron, to grow and then dissect Arabidopsis thaliana roots into three segments. We demonstrate that distinct endospheric and rhizosphere bacterial communities colonize the segments, supporting the hypothesis of microbiota differentiation along the axis. Root metabolite profiling of each segment reveals differential metabolite enrichment and specificity. Bioinformatic analyses and GUS histochemistry indicate microbe-induced accumulation of SWEET2, 4, and 12 sugar uniporters. Profiling of root segments from sweet mutants shows altered spatial metabolic profiles and reorganization of endospheric root microbiota. This work reveals the interdependency between root metabolites and microbial colonization and the contribution of SWEETs to spatial diversity and stability of microbial ecosystem.

Expanding the Toolbox for Genetic Manipulation in Pseudogymnoascus: RNAi-Mediated Silencing and CRISPR/Cas9-Mediated Disruption of a Polyketide Synthase Gene Involved in Red Pigment Production in P. verrucosus.

Fungi belonging to the genus Pseudogymnoascus have garnered increasing attention in recent years. One of the members of the genus, P. destructans, has been identified as the causal agent of a severe bat disease. Simultaneously, the knowledge of Pseudogymnoascus species has expanded, in parallel with the increased availability of genome sequences. Moreover, Pseudogymnoascus exhibits great potential as a producer of specialized metabolites, displaying a diverse array of biological activities. Despite these significant advancements, the genetic landscape of Pseudogymnoascus remains largely unexplored due to the scarcity of suitable molecular tools for genetic manipulation. In this study, we successfully implemented RNAi-mediated gene silencing and CRISPR/Cas9-mediated disruption in Pseudogymnoascus, using an Antarctic strain of Pseudogymnoascus verrucosus as a model. Both methods were applied to target azpA, a gene involved in red pigment biosynthesis. Silencing of the azpA gene to levels of 90% or higher eliminated red pigment production, resulting in transformants exhibiting a white phenotype. On the other hand, the CRISPR/Cas9 system led to a high percentage (73%) of transformants with a one-nucleotide insertion, thereby inactivating azpA and abolishing red pigment production, resulting in a white phenotype. The successful application of RNAi-mediated gene silencing and CRISPR/Cas9-mediated disruption represents a significant advancement in Pseudogymnoascus research, opening avenues for comprehensive functional genetic investigations within this underexplored fungal genus.

A comparison of four self-controlled study designs in an analysis of COVID-19 vaccines and myocarditis using five European databases.

INTRODUCTION: The aim of this study was to assess the possible extent of bias due to violation of a core assumption (event-dependent exposures) when using self-controlled designs to analyse the association between COVID-19 vaccines and myocarditis. METHODS: We used data from five European databases (Spain: BIFAP, FISABIO VID, and SIDIAP; Italy: ARS-Tuscany; England: CPRD Aurum) converted to the ConcePTION Common Data Model. Individuals who experienced both myocarditis and were vaccinated against COVID-19 between 1 September 2020 and the end of data availability in each country were included. We compared a self-controlled risk interval study (SCRI) using a pre-vaccination control window, an SCRI using a post-vaccination control window, a standard SCCS and an extension of the SCCS designed to handle violations of the assumption of event-dependent exposures. RESULTS: We included 1,757 cases of myocarditis. For analyses of the first dose of the Pfizer vaccine, to which all databases contributed information, we found results consistent with a null effect in both of the SCRI and extended SCCS, but some indication of a harmful effect in a standard SCCS. For the second dose, we found evidence of a harmful association for all study designs, with relatively similar effect sizes (SCRI pre = 1.99, 1.40 - 2.82; SCRI post 2.13, 95 %CI - 1.43, 3.18; standard SCCS 1.79, 95 %CI 1.31 - 2.44, extended SCCS 1.52, 95 %CI = 1.08 - 2.15). Adjustment for calendar time did not change these conclusions. Findings using all designs were also consistent with a harmful effect following a second dose of the Moderna vaccine. CONCLUSIONS: In the context of the known association between COVID-19 vaccines and myocarditis, we have demonstrated that two forms of SCRI and two forms of SCCS led to largely comparable results, possibly because of limited violation of the assumption of event-dependent exposures.