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BackgroundAccurate detection of severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) is necessary to mitigate the coronavirus disease-19 (COVID-19) pandemic. However, the test reagents and assay platforms are varied and may not be sufficiently robust to diagnose COVID-19.MethodsWe reviewed 85 studies (21,530 patients), published from five regions of the world, to highlight issues involved in the diagnosis of COVID-19 in the early phase of the pandemic, following the standards outlined in the PRISMA statement. All relevant articles, published up to May 31, 2020, in PubMed, BioRiXv, MedRiXv, and Google Scholar, were included. We evaluated the qualitative (9749 patients) and quantitative (10,355 patients) performance of RT-PCR and serologic diagnostic tests for real-world samples, and assessed the concordance (5,538 patients) between methods in meta-analyses.ResultsThe RT-PCR tests exhibited heterogeneity in the primers and reagents used. Of 1,957 positive RT-PCR COVID-19 participants, 1,585 had positive serum antibody (IgM +/- IgG) tests (sensitivity 0.81, 95%CI 0.66-.90). While 3,509 of 3581 participants RT-PCR negative for COVID-19 were found negative by serology testing (specificity 0.98, 95%CI 0.94-0.99). The chemiluminescent immunoassay exhibited the highest sensitivity, followed by ELISA and lateral flow immunoassays. Serology tests had higher sensitivity and specificity for laboratory-approval than for real-world reporting data.ConclusionsThe robustness of the assays/platforms is influenced by variability in sampling and reagents. Serological testing complements and may minimize false negative RT-PCR results. Lack of standardized assay protocols in the early phase of pandemic might have contributed to the spread of COVID-19.

Atypical Gaucher disease is caused by variants in the PSAP gene. Saposin C is one of four homologous proteins derived from sequential cleavage of the saposin precursor protein, prosaposin. It is an essential activator for glucocerebrosidase, which is deficient in Gaucher disease. Although atypical Gaucher disease due to deficiency of saposin C is rare, it exhibits vast phenotypic heterogeneity. Here, we report on a Pakistani family that exhibits features of Gaucher disease, i.e., prelingual profound sensorineural hearing impairment, vestibular dysfunction, hepatosplenomegaly, kyphosis, and thrombocytopenia. The family was investigated using exome and Sanger sequencing. A homozygous missense variant c.1076A>C: p.(Glu359Ala) in exon 10 of the PSAP gene was observed in all affected family members. In conclusion, we identified a new likely pathogenic missense variant in PSAP in a large consanguineous Pakistani family with atypical Gaucher disease. Gaucher disease due to a deficiency of saposin C has not been previously reported within the Pakistani population. Genetic screening of patients with the aforementioned phenotypes could ensure adequate follow-up and the prevention of further complications. Our finding expands the genetic and phenotypic spectrum of atypical Gaucher disease due to a saposin C deficiency.

BackgroundAccurate detection of SARS-CoV-2 is necessary to mitigate the COVID-19 pandemic. However, the test reagents and assay platforms are varied and may not be sufficiently robust to diagnose COVID-19.MethodsWe reviewed 85 studies (21 530 patients), published from five regions of the world, to highlight issues involved in the diagnosis of COVID-19 in the early phase of the pandemic. All relevant articles, published up to 31 May 2020, in PubMed, BioRiXv, MedRiXv and Google Scholar, were included. We evaluated the qualitative (9749 patients) and quantitative (10 355 patients) performance of RT-PCR and serologic diagnostic tests for real-world samples, and assessed the concordance (5538 patients) between test performance in meta-analyses. Synthesis of results was done using random effects modelling and bias was evaluated according to QUADAS-2 guidelines.ResultsThe RT-PCR tests exhibited heterogeneity in the primers and reagents used. Of 1957 positive RT-PCR COVID-19 participants, 1585 had positive serum antibody (IgM±IgG) tests (sensitivity 0.81, 95% CI 0.66 to 0.90). While 3509 of 3581 participants RT-PCR negative for COVID-19 were found negative by serology testing (specificity 0.98, 95% CI 0.94 to 0.99). The chemiluminescent immunoassay exhibited the highest sensitivity, followed by ELISA and lateral flow immunoassays. Serology tests had higher sensitivity and specificity for laboratory approval than for real-world reporting data.DiscussionThe robustness of the assays/platforms is influenced by variability in sampling and reagents. Serological testing complements and may minimise false negative RT-PCR results. Lack of standardised assay protocols in the early phase of pandemic might have contributed to the spread of COVID-19.

Background Atrial fibrillation (AF) is associated with a high risk of stroke and mortality. Some years ago, the EURObservational Research Programme launched the General Long-Term Registry with the aim to evaluate contemporary management of AF patients in Europe, the current use of vitamin K antagonists (VKAs), direct-acting oral anticoagulants (DOACs) and other AF treatments, in relation to guideline recommendations. Purpose The present report aims to describe the characteristics of a large database on the management of AF in Spain, using the Spanish cohort included in the EORP-AF Long-Term Registry. Methods The EORP-AF Long-Term General Registry is a prospective, observational, large-scale multicentre registry sponsored and conducted by the ESC, enrolling AF patients in current cardiology practices in 250 centres from 27 participating ESC countries. Patients were enrolled consecutively when presenting with AF as primary or secondary diagnosis to inpatients and outpatient cardiology services from October 2013 to September 2016. The first Spanish patient in the EORP-AF Long-Term Registry was included in 2014. Initially, the aim was to carry out a follow-up up to 3 years but this was reduced to 2 years by the Executive Committee. To date, only data from the first year of follow-up is available for the Spanish cohort. Results A cohort of 729 AF Spanish patients was included (57.1% male, median age 75 [IQR 67–81] years, median CHA2DS2-VASc and HAS-BLED of 3 [IQR 2–5] and 2 [IQR 1–2], respectively). A relatively low proportion of patients (634, 87%) received oral anticoagulants (OACs), of which 389 (53.4%) were on VKAs and 245 (33.6%) were on DOACs (rate ratio = 1.59 [95% CI 1.35–1.87], p Conclusions Baseline data of the Spanish cohort are similar to that reported for the whole EORP cohort, including similar stroke and bleeding risks. OAC use slightly increased at 1-year, with low discontinuation rates which could be related with a low incidence of thromboembolic events. However, despite the ∼8% rate of major bleeding in overall, the use of a safer therapy such as DOACs is still low compared to VKAs, being the antiplatelets commonly used concomitantly with OACs Funding Acknowledgement Type of funding source: Private grant(s) and/or Sponsorship. Main funding source(s): Unconditional grant by Boehringer-Ingelheim

Pseudoexon (PE) inclusion has been implicated in various dystrophinopathies, however, its splicing characteristics have not been fully investigated. This study aims to analyze the splicing characteristics of dystrophin PEs and compare them with those of dystrophin canonical exons (CEs). Forty-two reported dystrophin PEs were divided into a splice site (ss) group and a splicing regulatory element (SRE) group. Five dystrophin PEs with characteristics of poison exons were identified and categorized as the possible poison exon group. The comparative analysis of each essential splicing signal among different groups of dystrophin PEs and dystrophin CEs revealed that the possible poison exon group had a stronger 3&prime, ss compared to any other group. As for auxiliary SREs, different groups of dystrophin PEs were found to have a smaller density of diverse types of exonic splicing enhancers and a higher density of several types of exonic splicing silencers compared to dystrophin CEs. In addition, the possible poison exon group had a smaller density of 3&prime, ss intronic splicing silencers compared to dystrophin CEs. To our knowledge, our findings indicate for the first time that poison exons might exist in DMD (the dystrophin gene) and present with different splicing characteristics than other dystrophin PEs and CEs.

DNA samples from five members of a multiplex non-consanguineous Cameroonian family, segregating prelingual and progressive autosomal recessive non-syndromic sensorineural hearing impairment, underwent whole exome sequencing. We identified novel bi-allelic compound heterozygous pathogenic variants in CLIC5. The variants identified, i.e., the missense [NM_016929.5:c.224T&gt, C, p.(L75P)] and the splicing (NM_016929.5:c.63+1G&gt, A), were validated using Sanger sequencing in all seven available family members and co-segregated with hearing impairment (HI) in the three hearing impaired family members. The three affected individuals were compound heterozygous for both variants, and all unaffected individuals were heterozygous for one of the two variants. Both variants were absent from the genome aggregation database (gnomAD), the Single Nucleotide Polymorphism Database (dbSNP), and the UK10K and Greater Middle East (GME) databases, as well as from 122 apparently healthy controls from Cameroon. We also did not identify these pathogenic variants in 118 unrelated sporadic cases of non-syndromic hearing impairment (NSHI) from Cameroon. In silico analysis showed that the missense variant CLIC5-p.(L75P) substitutes a highly conserved amino acid residue (leucine), and is expected to alter the stability, the structure, and the function of the CLIC5 protein, while the splicing variant CLIC5-(c.63+1G&gt, A) is predicted to disrupt a consensus donor splice site and alter the splicing of the pre-mRNA. This study is the second report, worldwide, to describe CLIC5 involvement in human hearing impairment, and thus confirms CLIC5 as a novel non-syndromic hearing impairment gene that should be included in targeted diagnostic gene panels.

DNA samples from five members of a multiplex non-consanguineous Cameroonian family, segregating prelingual and progressive autosomal recessive non-syndromic sensorineural hearing impairment, underwent whole exome sequencing. We identified novel bi-allelic compound heterozygous pathogenic variants in CLIC5. The variants identified, i.e., the missense [NM_016929.5:c.224T>C; p.(L75P)] and the splicing (NM_016929.5:c.63+1G>A), were validated using Sanger sequencing in all seven available family members and co-segregated with hearing impairment (HI) in the three hearing impaired family members. The three affected individuals were compound heterozygous for both variants, and all unaffected individuals were heterozygous for one of the two variants. Both variants were absent from the genome aggregation database (gnomAD), the Single Nucleotide Polymorphism Database (dbSNP), and the UK10K and Greater Middle East (GME) databases, as well as from 122 apparently healthy controls from Cameroon. We also did not identify these pathogenic variants in 118 unrelated sporadic cases of non-syndromic hearing impairment (NSHI) from Cameroon. In silico analysis showed that the missense variant CLIC5-p.(L75P) substitutes a highly conserved amino acid residue (leucine), and is expected to alter the stability, the structure, and the function of the CLIC5 protein, while the splicing variant CLIC5-(c.63+1G>A) is predicted to disrupt a consensus donor splice site and alter the splicing of the pre-mRNA. This study is the second report, worldwide, to describe CLIC5 involvement in human hearing impairment, and thus confirms CLIC5 as a novel non-syndromic hearing impairment gene that should be included in targeted diagnostic gene panels.

Otitis media (OM), a very common disease in young children, can result in hearing loss. In order to potentially replicate previously reported associations between OM and PLG, exome and Sanger sequencing, RNA-sequencing of saliva and middle ear samples, 16S rRNA sequencing, molecular modeling, and statistical analyses including transmission disequilibrium tests (TDT) were performed in a multi-ethnic cohort of 718 families and simplex cases with OM. We identified four rare PLG variants c.112A > G (p.Lys38Glu), c.782G > A (p.Arg261His), c.1481C > T (p.Ala494Val) and c.2045 T > A (p.Ile682Asn), and one common variant c.1414G > A (p.Asp472Asn). However TDT analyses for these PLG variants did not demonstrate association with OM in 314 families. Additionally PLG expression is very low or absent in normal or diseased middle ear in mouse and human, and salivary expression and microbial α-diversity were non-significant in c.1414G > A (p.Asp472Asn) carriers. Based on molecular modeling, the novel rare variants particularly c.782G > A (p.Arg261His) and c.2045 T > A (p.Ile682Asn) were predicted to affect protein structure. Exploration of other potential disease mechanisms will help elucidate how PLG contributes to OM susceptibility in humans. Our results underline the importance of following up findings from genome-wide association through replication studies, preferably using multi-omic datasets.

This work investigates the performance of the on-demand machine learning (ODML) algorithm introduced in Leal et al. (Transp. Porous Media 133(2), 161-204, 2020) when applied to different reactive transport problems in heterogeneous porous media. This approach was devised to accelerate the computationally expensive geochemical reaction calculations in reactive transport simulations. We demonstrate that even with a strong heterogeneity present, the ODML algorithm speeds up these calculations by one to three orders of magnitude. Such acceleration, in turn, significantly advances the entire reactive transport simulation. The performed numerical experiments are enabled by the novel coupling of two open-source software packages: Reaktoro (Leal 2015) and Firedrake (Rathgeber et al. ACM Trans. Math. Softw. 43(3), 2016). The first library provides the most recent version of the ODML approach for the chemical equilibrium calculations, whereas, the second framework includes the newly implemented conservative Discontinuous Galerkin finite element scheme for the Darcy problem, i.e., the Stabilized Dual Hybrid Mixed (SDHM) method Ntifiez et al. (Int. J. Model. Simul. Petroleum Industry, 6, 2012)., Computational Geosciences, 26, ISSN:1420-0597, ISSN:1573-1499

We performed in vitro susceptibility testing for eravacycline in comparison to 4 other antimicrobials against 10 Mycoplasma genitalium , 40 Mycoplasma hominis , 44 Mycoplasma pneumoniae , 20 Ureaplasma parvum , and 20 Ureaplasma urealyticum isolates. All eravacycline MICs were ≤0.25 μg/ml, except that for one isolate of M. genitalium , for which the MIC was 2 μg/ml.

Congenital hearing impairment is a sensory disorder that is genetically highly heterogeneous. By performing exome sequencing in two families with congenital nonsyndromic profound sensorineural hearing loss (SNHL), we identified autosomal dominantly inherited missense variants [p.(Asn283Ser), p.(Thr116Ile)] in GREB1L, a neural crest regulatory molecule. The p.(Thr116Ile) variant was also associated with bilateral cochlear aplasia and cochlear nerve aplasia upon temporal bone imaging, an ultra-rare phenotype previously seen in patients with de novo GREB1L variants. An important role of GREB1L in normal ear development has also been demonstrated by greb1l&minus, /&minus, zebrafish, which show an abnormal sensory epithelia innervation. Last, we performed a review of all disease-associated variation described in GREB1L, as it has also been implicated in renal, bladder and genital malformations. We show that the spectrum of features associated with GREB1L is broad, variable and with a high level of reduced penetrance, which is typically characteristic of neurocristopathies. So far, seven GREB1L variants (14%) have been associated with ear-related abnormalities. In conclusion, these results show that autosomal dominantly inherited variants in GREB1L cause profound SNHL. Furthermore, we provide an overview of the phenotypic spectrum associated with GREB1L variants and strengthen the evidence of the involvement of GREB1L in human hearing.

Cirrhotic cardiomyopathy is characterized by the presence of structural and functional cardiac alterations in patients suffering from hepatic cirrhosis, without previously known cardiac causes that may explain it. Clinically, it is characterized by the presence of variable grades of diastolic and systolic dysfunction, alterations in the electric conductance (elongation of corrected QT interval) and inadequate chronotropic response. This pathology has been related to substandard response in the management of patients with portal hypertension and poor outcome after transplant. Even when the first description of this pathology dates back from 1953, it remains a poorly studied and frequently underdiagnosed entity. Echocardiography prevails as a practical diagnostic tool for this pathology since simple measurements as the E/A index can show diastolic dysfunction. Systolic dysfunction discloses as a diminished ejection fraction of the left ventricle and the latent forms are detected by echocardiography studies with pharmacological stress. In recent years new techniques such as the longitudinal strain have been studied and they seem promising for the detection of early alterations.La miocardiopatía cirrótica se caracteriza por la presencia de alteraciones cardiacas estructurales y funcionales en pacientes con cirrosis hepática, sin que existan otras causas de enfermedad cardiaca. Clínicamente se caracteriza por la presencia de grados variables de disfunción diastólica y sistólica, alteraciones de la conducción eléctrica (prolongación del intervalo QT) y respuesta cronotrópica inapropiada. Esta patología se ha relacionado con desenlaces clínicos adversos, mala respuesta en el manejo de la hipertensión portal y resultados desfavorables posterior a trasplante hepático ortotópico. A pesar de que las primeras descripciones datan de 1953, es una entidad poco estudiada y frecuentemente subdiagnosticada. El ecocardiograma es una herramienta de diagnóstico importante en esta entidad. Mediciones simples como el índice E/A pueden traducir disfunción diastólica. La disfunción sistólica se manifiesta con disminución de la fracción de eyección del ventrículo izquierdo y las formas latentes se detectan mediante estudios de ecocardiografía con estrés farmacológico; en los últimos años se han estudiado otras técnicas como el strain longitudinal, que parecen prometedoras en la detección de alteraciones tempranas.

Sarcina ventriculi is a gram-positive anaerobic bacterium reported rarely in patients with a history of gastrointestinal surgery and delayed gastric emptying. Sarcina has been implicated in the development of gastric ulcers, emphysematous gastritis, and gastric perforation. So far, less than 30 cases of Sarcina isolated from gastric specimens have been reported, including 3 cases associated with life-threatening illness: emphysematous gastritis and gastric perforation. Herein, we report a case of a 58-yearold woman with history of gastric surgery who presented for evaluation of persistent gastric pain and incurable ulcer. She underwent total gastrectomy, and the resected stomach demonstrated a perforated ulcer with the presence of Sarcina microorganisms. We also report a second case of a 56-year-old woman with history of NSAID use who presented with gastric outlet obstruction. The gastric biopsy identifi ed concurrent Helicobacter pylori and Sarcina. Given Sarcina‘s association with emphysematous gastritis and gastric perforation, its identifi cation on gastric biopsies should be clearly stated in pathology reports and, depending on the clinical scenario, prompt clinicians to add adjunctive antimicrobials to anti-ulcer therapeutic regimens.

SUMMARY Recently developed approaches for highly multiplexed imaging have revealed complex patterns of cellular positioning and cell-cell interactions with important roles in both cellular- and tissue-level physiology. However, tools to quantitatively study cellular patterning and tissue architecture are currently lacking. Here, we develop a spatial analysis toolbox, the histo-cytometric multidimensional analysis pipeline (CytoMAP), which incorporates data clustering, positional correlation, dimensionality reduction, and 2D/3D region reconstruction to identify localized cellular networks and reveal features of tissue organization. We apply CytoMAP to study the microanatomy of innate immune subsets in murine lymph nodes (LNs) and reveal mutually exclusive segregation of migratory dendritic cells (DCs), regionalized compartmentalization of SIRPa dermal DCs, and preferential association of resident DCs with select LN vasculature. The findings provide insights into the organization of myeloid cells in LNs and demonstrate that CytoMAP is a comprehensive analytics toolbox for revealing features of tissue organization in imaging datasets., In Brief Stoltzfus et al. present CytoMAP, a spatial analytics platform that incorporates diverse statistical and visualization modules for analysis of cellular positioning, cell-cell interactions, global tissue structure, and heterogeneity of tissue microenvironments. Exploration of myeloid cell localization in lymph nodes reveals fundamental positional relationships between dendritic cell subsets and local vasculature., Graphical Abstract

To determine if the Primary Health care professionals prescribe physical exercise (PE) using adequate knowledge criteria.A comparative cross-sectional descriptive and observational study carried out in 32 health centres in the Region of Murcia. A total of 476 doctors and 327 nurses were included. A questionnaire was administered to all of them, in which the variables analysed were: Level of overall and specific knowledge by blocks of pathologies (lipids, diabetes, hypertension, obesity, and generalities) on the prescribing of PE to patients with cardiovascular risk, as well as related variables associated with the quality of prescribing PE.Statistically significant differences in knowledge were found in the group that recommends heart rate measurement in «30% to 70%» of their patients. Differences in the subgroups were observed in the sections on diabetes (P0.05), obesity (P 0.05), and hypertension (P0.05). When analysing the knowledge values obtained according to the proportion of patients to whom «very rarely» PE time is specified, significant differences are observed (P0.05), showing doctors with a higher level of knowledge. The differences were accentuated between both professional groups in the group of respondents who specify time to «less than 30%» of their patients (P0.01). There are no differences between those professionals who specify time to «more than 31%» of their patients, although the trend shows higher knowledge values among doctors.The prescription of physical exercise among Primary Care professionals is not carried out using adequate knowledge criteria.

We evaluated six commercial molecular tests targeting Mycoplasma pneumoniae, namely, the BioFire FilmArray respiratory panel (RP), the Meridian Alethia Mycoplasma Direct, the GenMark ePlex respiratory pathogen panel (RPP), the Luminex NxTAG RPP, the ELITech ELITe InGenius Mycoplasma MGB research use only (RUO) PCR, and the SpeeDx Resistance Plus MP assays. Laboratory-developed PCR assays at the University of Alabama at Birmingham and the Centers for Disease Control and Prevention were used as reference standards. Among 428 specimens, 212 were designated confirmed positives for M. pneumoniae. The highest clinical sensitivities were found with the InGenius PCR (99.5%) and the FilmArray RP (98.1%). The Resistance Plus MP identified 93.3% of the confirmed-positive specimens, whereas 83.6, 64.6, and 55.7% were identified by the ePlex RPP, NxTAG RPP, and Mycoplasma Direct assays, respectively. There was no significant difference between the sensitivity of the reference methods and that of the FilmArray RP and InGenius assays, but the remaining four assays detected significantly fewer positive specimens (P

Objectives Vaccines that prime Wilms' tumor 1 (WT1)‐specific CD8+ T cells are attractive cancer immunotherapies. However, immunogenicity and clinical response rates may be enhanced by delivering WT1 to CD141+ dendritic cells (DCs). The C‐type lectin‐like receptor CLEC9A is expressed exclusively by CD141+ DCs and regulates CD8+ T‐cell responses. We developed a new vaccine comprising a human anti‐CLEC9A antibody fused to WT1 and investigated its capacity to target human CD141+ DCs and activate naïve and memory WT1‐specific CD8+ T cells. Methods WT1 was genetically fused to antibodies specific for human CLEC9A, DEC‐205 or β‐galactosidase (untargeted control). Activation of WT1‐specific CD8+ T‐cell lines following cross‐presentation by CD141+ DCs was quantified by IFNγ ELISPOT. Humanised mice reconstituted with human immune cell subsets, including a repertoire of naïve WT1‐specific CD8+ T cells, were used to investigate naïve WT1‐specific CD8+ T‐cell priming. Results The CLEC9A‐WT1 vaccine promoted cross‐presentation of WT1 epitopes to CD8+ T cells and mediated priming of naïve CD8+ T cells more effectively than the DEC‐205‐WT1 and untargeted control‐WT1 vaccines. Conclusions Delivery of WT1 to CD141+ DCs via CLEC9A stimulates CD8+ T cells more potently than either untargeted delivery or widespread delivery to all Ag‐presenting cells via DEC‐205, suggesting that cross‐presentation by CD141+ DCs is sufficient for effective CD8+ T‐cell priming in humans. The CLEC9A‐WT1 vaccine is a promising candidate immunotherapy for malignancies that express WT1., This project developed a new vaccine comprising a human anti‐CLEC9A antibody fused to the immunogenic, broadly expressed tumor antigen, WT1. This vaccine specifically delivered WT1 to human CD141+ (cDC1) dendritic cells, for efficient cross‐presentation and activation of naïve and memory WT1‐specific CD8+ T cells. The anti‐CLEC9A‐WT1 antibody is a promising new vaccine candidate for malignancies that express WT1.

BACKGROUND: Otitis media (OM) susceptibility has significant heritability, however the role of rare variants in OM is mostly unknown. Our goal is to identify novel rare variants that confer OM susceptibility. METHODS: We performed exome and Sanger sequencing of >1,000 DNA samples from 551 multi-ethnic families with OM and unrelated individuals, RNA-sequencing, and microbiome sequencing and analyses of swabs from the outer ear, middle ear, nasopharynx and oral cavity. We also examined protein localization and gene expression in infected and healthy middle ear tissues. RESULTS: A large, intermarried pedigree that includes 81 OM-affected and 53 unaffected individuals co-segregates two known rare A2ML1 variants, a common FUT2 variant and a rare, novel pathogenic variant c.1682A>G (p.Glu561Gly) within SPINK5 (LOD=4.09). Carriage of the SPINK5 missense variant resulted in increased relative abundance of Microbacteriaceae in the middle ear, along with occurrence of Microbacteriaceae in the outer ear and oral cavity but not the nasopharynx. Eight additional novel SPINK5 variants were identified in twelve families and individuals with OM. A role for SPINK5 in OM susceptibility is further supported by lower RNA counts in variant carriers, strong SPINK5 localization in outer ear skin, faint localization to middle ear mucosa and eardrum, and increased SPINK5 expression in human cholesteatoma. CONCLUSION: SPINK5 variants confer susceptibility to non-syndromic OM. These variants potentially contribute to middle ear pathology through breakdown of mucosal and epithelial barriers, immunodeficiency such as poor vaccination response, alteration of head and neck microbiota, and facilitation of entry of opportunistic pathogens into the middle ear.

Peanut is a crop of the Kayabi tribe, inhabiting the Xingu Indigenous Park, Brazil. Morphological analysis of Xingu accessions showed variation exceeding that described for cultivated peanuts. This raised questions as to the origin of the Xingu accessions: are they derived from different species, or is their diversity a result of different evolutionary and selection processes? To answer these questions, cytogenetic and genotyping analyses were conducted. The karyotypes of Xingu accessions analyzed are very similar to each other, to an A. hypogaea subsp. fastigiata accession and to the wild allotetraploid A. monticola. The accessions share the number and general morphology of the chromosomes; DAPI+ bands; 5S and 45S rDNA loci distribution and a high genomic affinity with A. duranensis and A. ipaënsis genomic probes. However, the number of CMA3 + bands differs from those determined for A. hypogaea and A. monticola, which are also different from each other. SNP genotyping grouped all Arachis allotetraploids into four taxonomic groups: Xingu accessions were closer to A. monticola and A. hypogaea subsp. hypogaea. Our data suggests that the morphological diversity within these accessions is not associated with a different origin and can be attributed to morphological plasticity and different selection by the Indian tribes.

Este artículo se encuentra disponible en la siguiente URL: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0228916 Objectives This study sought to develop a tool for evaluating person-centered therapeutic relationships within physiotherapy services, and to examine the content validity of the same. Methods A mixed qualitative and quantitative study was performed in three distinct phases: 1) the items were generated based on a literature review and a content analysis of focus groups of patients and physiotherapists; 2) an e-Delphi survey process was performed based on three rounds to select and refine the proposed questionnaire; 3) two rounds of cognitive interviews were conducted to evaluate the comprehension of items, the clarity of language and the appropriateness and relevance of content. Results Thirty-one items were generated based on the seven domains identified after the analysis of four focus groups of physiotherapists and four patient focus groups. Nine experts participated in the e-Delphi survey. Fifty-five patients participated in the two rounds of the cognitive pre-tests. Participating patients were from public and private physical therapy services. Based on the participants’ suggestions, four items were removed, and four were added, whereas 16 were reworded. Conclusions The final tool comprised 31 items divided into seven domains. The response format was based on a 5-point Likert frequency scale. The response options ranged from “strongly agree” to “strongly disagree”.

Objectives\ud Vaccines that prime Wilms' tumor 1 (WT1)‐specific CD8+ T cells are attractive cancer immunotherapies. However, immunogenicity and clinical response rates may be enhanced by delivering WT1 to CD141+ dendritic cells (DCs). The C‐type lectin‐like receptor CLEC9A is expressed exclusively by CD141+ DCs and regulates CD8+ T‐cell responses. We developed a new vaccine comprising a human anti‐CLEC9A antibody fused to WT1 and investigated its capacity to target human CD141+ DCs and activate naïve and memory WT1‐specific CD8+ T cells.\ud \ud Methods\ud WT1 was genetically fused to antibodies specific for human CLEC9A, DEC‐205 or β‐galactosidase (untargeted control). Activation of WT1‐specific CD8+ T‐cell lines following cross‐presentation by CD141+ DCs was quantified by IFNγ ELISPOT. Humanised mice reconstituted with human immune cell subsets, including a repertoire of naïve WT1‐specific CD8+ T cells, were used to investigate naïve WT1‐specific CD8+ T‐cell priming.\ud \ud Results\ud The CLEC9A‐WT1 vaccine promoted cross‐presentation of WT1 epitopes to CD8+ T cells and mediated priming of naïve CD8+ T cells more effectively than the DEC‐205‐WT1 and untargeted control‐WT1 vaccines.\ud \ud Conclusions\ud Delivery of WT1 to CD141+ DCs via CLEC9A stimulates CD8+ T cells more potently than either untargeted delivery or widespread delivery to all Ag‐presenting cells via DEC‐205, suggesting that cross‐presentation by CD141+ DCs is sufficient for effective CD8+ T‐cell priming in humans. The CLEC9A‐WT1 vaccine is a promising candidate immunotherapy for malignancies that express WT1.

Sinoatrial node dysfunction and deafness (SANDD) syndrome is rare and characterized by a low heart beat and severe-to-profound deafness. Additional features include fatigue, dizziness, and episodic syncope. The sinoatrial node (SAN) drives heart automaticity and continuously regulates heart rate. The CACNA1D gene encoding the Ca(v)1.3 protein expressed in inner hair cells, atria and SAN, induces loss-of-function in channel activity and underlies SANDD. To date, only one variant c.1208_1209insGGG:p.(G403_V404insG) has been reported for SANDD syndrome. We studied five Pakistani families with SANDD and characterized a new missense variant p.(A376V) in CACNA1D in one family, and further characterized the founder variant p.(G403_V404insG) in four additional pedigrees. We show that affected individuals in the four families which segregate p.(G403_V404insG) share a 1.03 MB haplotype on 3p21.1 suggesting they share a common distant ancestor. In conclusion, we identified new and known variants in CACNA1D in five Pakistani families with SANDD. This study is of clinical importance as the CACNA1D founder variant is only observed in families from the Khyber Pakhtunkhwa (KPK) province, in Pakistan. Therefore, screening patients with congenital deafness for SAN dysfunction in this province could ensure adequate follow-up and prevent cardiac failure associated with SAN.

Consanguineous Pakistani pedigrees segregating deafness have contributed decisively to the discovery of 31 of the 68 genes associated with nonsyndromic autosomal recessive hearing loss (HL) worldwide. In this study, we utilized genome-wide genotyping, Sanger and exome sequencing to identify 163 DNA variants in 41 previously reported HL genes segregating in 321 Pakistani families. Of these, 70 (42.9%) variants identified in 29 genes are novel. As expected from genetic studies of disorders segregating in consanguineous families, the majority of affected individuals (94.4%) are homozygous for HL-associated variants, with the other variants being compound heterozygotes. The five most common HL genes in the Pakistani population are SLC26A4, MYO7A, GJB2, CIB2 and HGF, respectively. Our study provides a profile of the genetic etiology of HL in Pakistani families, which will allow for the development of more efficient genetic diagnostic tools, aid in accurate genetic counseling, and guide application of future gene-based therapies. These findings are also valuable in interpreting pathogenicity of variants that are potentially associated with HL in individuals of all ancestries. The Pakistani population, and its infrastructure for studying human genetics, will continue to be valuable to gene discovery for HL and other inherited disorders.

Polydactyly is a common congenital anomaly of the hand and foot. Postaxial polydactyly (PAP) is characterized by one or more posterior or postaxial digits. In a Pakistani family with autosomal recessive nonsyndromic postaxial polydactyly type A (PAPA), we performed genomewide genotyping, linkage analysis, and exome and Sanger sequencing. Exome sequencing revealed a homozygous nonsense variant (c.478C>T, p.[Arg160*]) in the FAM92A gene within the mapped region on 8q21.13-q24.12 that segregated with the PAPA phenotype. We found that FAM92A is expressed in the developing mouse limb and E11.5 limb bud including the progress zone and the apical ectodermal ridge, where it strongly localizes at the cilia level, suggesting an important role in limb patterning. The identified variant leads to a loss of the FAM92A/Chibby1 complex that is crucial for ciliogenesis and impairs the recruitment and the colocalization of FAM92A with Chibby1 at the base of the cilia. In addition, we show that Fam92a-/- homozygous mice also exhibit an abnormal digit morphology, including metatarsal osteomas and polysyndactyly, in addition to distinct abnormalities on the deltoid tuberosity of their humeri. In conclusion, we present a new nonsyndromic PAPA ciliopathy due to a loss-of-function variant in FAM92A. © 2018 American Society for Bone and Mineral Research.

Introduction: Dementia units (DU) provide comprehensive and specialised care to patients with dementia. However, assessment in these units normally focuses on patient management. The aim of this study was to determine satisfaction of the caregivers of patients managed in a DU and how they rated DU care. Methods: We created a self-administered questionnaire which was completed by 236 caregivers visiting the DU in 2015. The questionnaire included 6 dimensions (accessibility, organisation, professionalism, relationship with staff, information, and facilities); data were analysed using problem rates. Results: A total of 53.4% caregivers completed the questionnaire; most were women, patients’ spouses, or first-degree relatives. The overall problem rates was 15.0% (95% CI, 13.9-16.1), with sizeable differences between dimensions: from 0.1% (95% CI, 0.0-0.4) for ‘relationship with staff’ and ‘professionalism’ to 49.3% (95% CI, 45.4-53.2) for ‘information’. Waiting times over 30 minutes were perceived as excessive. Information problems were not directly related to patient management. Satisfaction was scored 8.29/10 (median 8; SD 1.45); 77.3% (95% CI, 69.8-74.8) of respondents were highly satisfied. Ninety-nine percent of the caregivers stated that they would recommend the DU. Conclusions: This questionnaire provides interesting data on care quality as perceived by patients’ caregivers. Our results have allowed us to identify problems and implement corrective actions. Our questionnaire has proved to be a useful tool for evaluating and improving care quality in DUs. Resumen: Introducción: Las unidades de demencia (UD) realizan un abordaje integral y especializado de las personas con demencia. Su evaluación suele limitarse a aspectos asistenciales. El objetivo del trabajo fue conocer la satisfacción y calidad percibida por familiares/cuidadores de pacientes atendidos por una UD. Métodos: Encuesta autocumplimentada con cuestionario de elaboración propia a 236 acompañantes de pacientes ya conocidos en la UD durante 2015, estructurada en 6 dimensiones (accesibilidad, organización, profesionalidad, trato, información e infraestructura) y analizada mediante tasas de problemas. Resultados: Tasa de respuesta de 53,4%, resultando la mayoría mujeres, cónyuges o familiares de primer grado de los pacientes. La tasa de problemas global fue 15,0% (IC 95%: 13,9-16,1), con amplias variaciones por dimensión: desde 0,1% (IC 95%: 0,0-0,4) en «trato» y «profesionalidad» hasta 49,3% (IC 95%: 45,4-53,2) en «información». La espera se percibe excesiva a partir de 30 min. Los problemas de información se centraron en actividades no directamente asistenciales. La satisfacción fue 8,29/10 (mediana 8; DE: 1,45) con 77,3% (IC 95%: 69,8-74,8) de puntuaciones excelentes. El 99% recomendarían la UD. Conclusiones: El cuestionario ha ofrecido una visión global de la calidad percibida por los familiares/cuidadores de los pacientes atendidos, y permitido identificar y priorizar dónde implantar acciones correctoras. Ha demostrado ser una herramienta útil para evaluar y mejorar la calidad asistencial en las UD. Keywords: Dementia, Perceived quality, Patient satisfaction, Questionnaires, Caregiver, Health organisations, Palabras clave: Demencia, Calidad percibida, Satisfacción del paciente, Cuestionarios, Cuidador, Organizaciones en salud

LHFPL5, the gene for DFNB67, underlies autosomal recessive nonsyndromic hearing impairment. We identified seven Pakistani families that mapped to 6p21.31, which includes the LHFPL5 gene. Sanger sequencing of LHFPL5 using DNA samples from hearing impaired and unaffected members of these seven families identified four variants. Among the identified variants, two were novel: one missense c.452 G > T (p.Gly151Val) and one splice site variant (c.*16 + 1 G > A) were each identified in two families. Two known variants: c.250delC (p.Leu84*) and c.380 A > G (p.Tyr127Cys) were also observed in two families and a single family, respectively. Nucleotides c.452G and c.*16 + 1G and amino-acid residue p.Gly151 are under strong evolutionary conservation. In silico bioinformatics analyses predicted these variants to be damaging. The splice site variant (c.*16 + 1 G > A) is predicted to affect pre-mRNA splicing and a loss of the 5' donor splice site in the 3'-untranslated region (3'-UTR). Further analysis supports the activation of a cryptic splice site approximately 357-bp downstream, leading to an extended 3'-UTR with additional regulatory motifs. In conclusion, we identified two novel variants in LHFPL5, including a unique 3'-UTR splice site variant that is predicted to impact pre-mRNA splicing and regulation through an extended 3'-UTR.

Molecular tests to diagnose conditions involving the disruption of normal microbiota are difficult to optimize. Using Nugent-scored Gram stain (NS) as the reference standard, we evaluated the performance of 3 molecular assays for the diagnosis of bacterial vaginosis (BV) and examined the impact of an incremental increase in bacterial targets. The BD Affirm assay includes a DNA probe for Gardnerella vaginalis, the Hologic transcription-mediated amplification (TMA) analyte-specific reagent (ASR) assay adds a second Lactobacillus sp. target, and the recently cleared in vitro diagnostic use (IVD) Aptima BV assay includes a third target (Atopobium vaginae). The diagnosis of vulvovaginal candidiasis (VVC) by the Affirm and Candida vaginitis Hologic TMA ASR assays was assessed using microscopy for yeast as the reference standard. From May to December 2018, 111 women with vaginitis symptoms prompting the clinician to order an Affirm test were enrolled with informed consent for the collection of additional specimens. Clinicians accurately predicted BV as the most likely diagnosis for 71% of the 45 patients with BV. Coinfection occurred in 13.5% of patients. For BV, the specificity of the Aptima IVD assay (86.3%) was higher than the Affirm assay (60.6%, P = 0.0002), but sensitivities were not significantly different. For VVC, the sensitivity of the ASR assay (100%) was higher than Affirm (75.9%; P = 0.023) and the specificity of the Affirm assay (98.8%) was higher than the ASR assay (86.6%; P = 0.004).

The rise in prices of the iron and manganese ores in the British market since the 1860s, triggered a real rush in several counties of the province of Alentejo, such as Santiago do Cacém (Cercal) and Odemira, with award of several concessions. With generally poor ores and lack of efficient and cheap transports, an iron-making attempt to increase profits by direct reduction was unsuccessfully carried at site, in the early1880s. However, the intention of local iron making persisted, as it appears from a royal charter of July 1892 that was not materialized. Thus, the lack of demand for these ores compromised local mining activity, which gradually declined until the end of the century., Na segunda metade do século XIX a procura e a alta de preços dos minérios de ferro e manganês no mercado britânico motivaram uma euforia mineira no Alentejo (sul de Portugal), com proliferação de concessões em concelhos como, entre outros, Santiago do Cacém (Cercal) e Odemira. Com minérios em geral pobres e sem transportes eficazes e baratos, a tentativa de valorização local da produção incluiu, na década de 1880, um malogrado ensaio de redução direta à boca da mina mantendo-se a intenção na década seguinte, explicitada num alvará de julho de 1892. Sem concretização e persistindo as limitações referidas, a ausência de procura levou ao definhar da atividade extrativa na região.

A new tetraglycosylated flavonoid derivative of kaempferol was isolated from the butanol extract of leaves of Platycyamus regnellii (Fabaceae) using high-speed countercurrent chromatography (HSCCC). The butanol extract (960.5 mg) was initially fractionated with the solvent system EtOAC-BuOH-H2O 2:8:10 (v/v/v) (elution in normal phase mode). The fractions containing the major tetraglycosylated flavonoid were further purified with the same solvent system by modifying the ratios to 3:7:10 (v/v/v) and alternating the elution to reversed phase mode. The structure of the isolated flavonoid (48.1 mg, 99.7% purity by ultra-high performance liquid chromatography with diode array detector (UHPLC-DAD)) was characterized by spectrometric and spectroscopic methods and identified as kaempferol 3-O-β-D-glucopiranosyl-(1®2)-α-L-rhamnopiranoside-(1®6)-α-L-rhamnopiranoside-7-O-α-L-rhamnopiranoside. Other minor flavonoids were also detected and tentatively identified using ultra-high performance liquid chromatography with diode array detector coupled to mass spectrometry using the electrospray ionization interface (UHPLC-DAD-ESI-MS/MS). By using the latter technique, a second tetraglycosylated flavonol with an O-methyl substituent (either rhamnetin, 3-O-methyl-quercetin or isorhamnetin) was detected, together with a triglycosylated kaempferol and diglycosylated flavonoids (either rhamnetin, 3-O-methyl-quercetin or isorhamnetin).

Resumen Objetivos Analizar la calidad de vida mediante el test SRS 22 en los pacientes con escoliosis de 20 o mas grados Cobb. Material y metodo Estudio descriptivo prospectivo realizado a pacientes con escoliosis de al menos 20 grados Cobb y con edad comprendida entre 10 y 20 anos. Fue realizado entre abril y mayo de 2016.Se les registro peso, talla, indice de masa corporal y cuestionario SR 22, que es un cuestionario especifico de calidad de vida para pacientes con escoliosis. Los pacientes se dividieron en 2 grupos para su analisis: a) escoliosis entre 20 y 29 grados Cobb (n = 44); y b) escoliosis de 30 grados Cobb o superior (n = 32). Resultados Existen diferencias significativas en las dimensiones que evaluan el dolor, la autopercepcion de la imagen y la satisfaccion del tratamiento, siendo peor valoradas cuando el grado de escoliosis es de 30 grados Cobb o superior. No hay diferencias significativas en la funcion/actividad ni en la salud mental. La puntuacion global del cuestionario tambien fue peor en el grupo con mayor grado de escoliosis. El peso, talla e indice de masa corporal no han mostrado diferencias significativas segun el mayor o menor grado de escoliosis. Conclusiones La escoliosis afecta de forma importante a la calidad de vida de las personas que la padecen, existiendo una correlacion negativa entre la gravedad de la escoliosis medida mediante grados Cobb y la calidad de vida.

Salicylic acid (SA) has a long history of safe use as ingredient in topical cosmetic products. In 2016, the Committee for Risk Assessment of the European Chemicals Agency proposed to classify SA as a Category 2 reproductive toxicant based on adverse developmental effects in animal toxicity studies. This hazard-based classification (based on mg/kg doses) requires a reassessment of the safety of the current SA concentrations in cosmetic consumer products. Herein, a safety reassessment was performed in which margins of safety were calculated based on literature data on the NOAEL plasma exposure levels from animal reproductive toxicity studies with ASA (rapidly converts to SA in plasma), human SA plasma levels from oral exposure to ASA and human dermal exposure to SA-containing cosmetic products. In addition, a literature review was performed, which shows that there are no adverse developmental effects despite extensive human clinical oral use of ASA up to the maximum recommended therapeutic doses. The plasma exposure-based safety assessment for SA combined with an absence of any clinical health risk with oral ASA use in the literature supports that there is an acceptable margin of safety for the consumer exposure to SA as authorized in the current EU cosmetic regulation.

Understanding patient perspectives regarding shared decision-making is crucial to providing informed, patient-centered care. Little is known about perceptions of vulnerable patients regarding shared decision-making during surgical consultation. The purpose of this study was to evaluate whether a validated tool reflects perceptions of shared decision-making accurately among patients seeking surgical consultation for gallstones at a safety-net hospital.A mixed methods study was conducted in a sample of adult patients with gallstones evaluated at a safety-net surgery clinic between May to July 2016. Semi-structured interviews were conducted after their initial surgical consultation and analyzed for emerging themes. Patients were administered the Shared Decision-Making Questionnaire and Autonomy Preference Scale. Univariate analyses were performed to identify factors associated with shared decision-making and to compare the results of the surveys to those of the interviews.The majority of patients (N = 30) were female (90%), Hispanic (80%), Spanish-speaking (70%), and middle-aged (45.7 ± 16 years). The proportion of patients who perceived shared decision-making was greater in the Shared Decision-Making Questionnaire versus the interviews (83% vs 27%, P .01). Age, sex, race/ethnicity, primary language, diagnosis, Autonomy Preference Scale score, and decision for operation was not associated with shared decision-making. Contributory factors to this discordance include patient unfamiliarity with shared decision-making, deference to surgeon authority, lack of discussion about different treatments, and confusion between aligned versus shared decisions.Available questionnaires may overestimate shared decision-making in vulnerable patients suggesting the need for alternative or modifications to existing methods. Furthermore, such metrics should be assessed for correlation with patient-reported outcomes, such as satisfaction with decisions and health status.

Peanut,Arachishypogaea(Linnaeus, 1753) is an allotetraploid cultivated plant with two subgenomes derived from the hybridization between two diploid wild species,A.duranensis(Krapovickas & W. C. Gregory, 1994) andA.ipaensis(Krapovickas & W. C. Gregory, 1994), followed by spontaneous chromosomal duplication. To understand genome changes following polyploidy, the chromosomes ofA.hypogaea, IpaDur1, an induced allotetraploid (A.ipaensis×A.duranensis)4xand the diploid progenitor species were cytogenetically compared. The karyotypes of the allotetraploids share the number and general morphology of chromosomes; DAPI+bands pattern and number of 5S rDNA loci. However, one 5S rDNA locus presents a heteromorphic FISH signal in both allotetraploids, relative to corresponding progenitor. Whilst forA.hypogaeathe number of 45S rDNA loci was equivalent to the sum of those present in the diploid species, in IpaDur1, two loci have not been detected. Overall distribution of repetitive DNA sequences was similar in both allotetraploids, althoughA.hypogaeahad additional CMA3+bands and few slight differences in the LTR-retrotransposons distribution compared to IpaDur1. GISH showed that the chromosomes of both allotetraploids had preferential hybridization to their corresponding diploid genomes. Nevertheless, at least one pair of IpaDur1 chromosomes had a clear mosaic hybridization pattern indicating recombination between the subgenomes, clear evidence that the genome of IpaDur1 shows some instability comparing to the genome ofA.hypogaeathat shows no mosaic of subgenomes, although both allotetraploids derive from the same progenitor species. For some reasons, the chromosome structure ofA.hypogaeais inherently more stable, or, it has been at least, partially stabilized through genetic changes and selection.

BACKGROUND: Autosomal recessive ichthyosis with hypotrichosis (ARIH; MIM 602400) syndrome is characterized by diffused congenital ichthyosis and generalized non-scarring hypotrichosis. The underlying genetic cause of ARIH syndrome has been associated with sequence variants of the gene ST14, encoding type II transmembrane serine protease matriptase, which maps to chromosome 11q24.3. OBJECTIVES: The current report aimed to investigate the clinical features and genetic cause of ARIH syndrome in a large consanguineous family of Pakistani origin. MATERIALS & METHODS: The technique of homozygosity mapping with highly polymorphic microsatellite markers was employed to establish linkage within the family. Sanger sequencing of exons and intron-exonboundaries of ST14 was performed to identify the potential pathogenic sequence variants, followed by structural analysis of the mutated protein. RESULTS: Linkage was established to chromosome 11q24.3, comprising the gene ST14. Sequence analysis led to the identification of a novel homozygous missense variant (c.1315G>A, p.Gly439Ser) in the ST14 gene that co-segregated with the disease phenotype in all affected members. Homology modelling and molecular docking analysis of ST14 with wild-type TMEFF1 protein was performed which revealed that glycine at position 439 is crucial for maintaining normal structural confirmation and interaction with the EGF domain of TMEFF1 protein. CONCLUSION: Taken together, the data strongly advocate this ST14 variant as the underlying genetic cause of ARIH syndrome in this first reported affected family from Pakistan. Moreover, the present study adds to the spectrum of mutations in the ST14 gene, implicating them in the pathogenesis of ARIH syndrome.

Mice reconstituted with human hematopoietic stem cells are valuable models to study aspects of the human immune system in vivo. We describe a humanized mouse model (hu mice) in which fully functional human CD141+ and CD1c+ myeloid and CD123+ plasmacytoid dendritic cells (DC) develop from human cord blood CD34+ cells in immunodeficient mice. CD141+ DC are the human equivalents of murine CD8+ /CD103+ DC which are essential for the induction of tumor-inhibitory cytotoxic T lymphocyte (CTL) responses, making them attractive targets to exploit for the development of new cancer immunotherapies. We used CD34+ -engrafted NSG-A2 mice to investigate activation of DC subsets by synthetic dsRNA or ssRNA analogues polyinosinic-polycytidylic acid/poly I:C and Resiquimod/R848, agonists for TLR3 and TLR8 respectively, both of which are expressed by CD141+ DC. Injection of hu mice with these agonists resulted in upregulation of costimulatory molecules CD80, CD83 and CD86 by CD141+ and CD1c+ DC alike, and their combination further enhanced expression of these molecules by both subsets. When combined, poly I:C and R848 enhanced serum levels of key cytokines associated with cross-presentation and the induction of CTL responses including IFN-α, IFN-β, IL-12 and CXCL10. These data advocate a combination of poly I:C and R848 TLR agonists as means of activating human DC for immunotherapy. This article is protected by copyright. All rights reserved.

Precursors of chelate pyridine–N-heterocyclic carbene (N^C:) ligands with methyl- or benzyl-substituted imidazolylidene fragments were synthesized. They were used to obtain 12 iridium bis-cyclometalated complexes of the type [Ir(C^N)2(N^C:)]+ (C^N = 2-(phenyl)pyridinato-C2,N, ppy, 2-(4,6-difluorophenyl)pyridinato-C2,N, dfppy). The ancillary N^C: ligands contain different structural modifications. The aim of the work was to analyze the effect that changes in the two types of ligands have on the photophysical and electrochemical properties and also on the behavior of these materials as photosensitizers. The X-ray crystal structures of five complexes were determined. The complexes emitted in the blue-green region. It was expected that the frontier orbitals and thus the photophysical and electrochemical properties would be controlled by the main C^N ligands, and it was demonstrated that the effect of the modifications in the N^C: ligand, especially the presence of a nitro group in the pyridine ring or the int...

The interaction of RNA and DNA with two aluminium-containing complexes has been studied at 25 °C using 1H-NMR, UV and CD spectroscopies, viscosity measurements and DFT simulations. The complex identified at pH = 7, Al13Cacx, can interact with polynucleotides by external binding. Even though the affinity of the aluminium complex for RNA and DNA is similar, the conformational changes induced upon interaction are greater with RNA. In contrast, at pH = 5 the complex present in solution is Al2Cac2, where the aluminium atoms have different configurations, such as octahedral (OH) and distorted square planar (DSP). Al2Cac2 is prone to interact covalently with both P (RNA or DNA) to yield Al2Cac2/P complexes. These complexes undergo slow decomposition in two consecutive steps to give Al2Cac1/P after losing the cacodylate moiety belonging to the Al(OH) and Al2Cac0/P when the second cacodylate residue of the Al(DSP) is lost. These steps are (roughly) twice as fast with DNA than with RNA and for both systems the release of the first cacodylate molecule is about 10 times faster than the second. The cacodylate present in the buffer acts not only as a complexing agent of Al but also as an active vehicle that leads the metal to its biological target; once the target is reached, the two cacodylate molecules are released, leaving Al covalently linked to the N7 of guanine and adenine. These features reveal the great complexity of aluminium interaction with biological systems, an important outcome due to the implications of this metal in human health.

Vegetation filters (VFs), a type of land application system, are a robust technology based on natural treatment mechanisms for the removal of wastewater contaminants. Their capacity to attenuate emerging organic contaminants (EOCs) has not yet been evaluated. The present study reports the results of a 2-year EOC monitoring carried out using a poplar VF receiving wastewater primarily treated by an Imhoff tank. The compounds selected included analgesics, a �-adrenergic blocker, stimulants, an anticonvulsant, an anti-depressant, an anti-inflammatory, an antibiotic and analgesic and stimulant metabolites. EOCs were analysed in the Imhoff tank effluent, in the infiltrated water at a depth of 90 cm and in the groundwater at a depth of 10 m. The results demonstrated that EOC attenuation was more significant in the first 90 cm than in the rest of the soil profile. The removal efficiency for all of the selected EOCs was higher than 90% with the exception of ketoprofen, which may pose a higher threat of groundwater contamination. The observed attenuation correlated with the hydrophobicity and charge state of the EOCs. The higher persistence of the metabolites 4-AAA and 4-FAA shows that progression in the degradation pathway does not always imply a mitigation of contamination.

A patient in Pennsylvania, USA, with common variable immunodeficiency sought care for fever, cough, and abdominal pain. Imaging revealed lesions involving multiple organs. Liver resection demonstrated necrotizing granulomas, recognizable tegument, and calcareous corpuscles indicative of an invasive cestode infection. Sequencing revealed 98% identity to a Versteria species of cestode found in mink.

A patient in Pennsylvania, USA, with common variable immunodeficiency sought care for fever, cough, and abdominal pain. Imaging revealed lesions involving multiple organs. Liver resection demonstrated necrotizing granulomas, recognizable tegument, and calcareous corpuscles indicative of an invasive cestode infection. Sequencing revealed 98% identity to a Versteria species of cestode found in mink.

Background The ePlex BCID Gram-Positive (GP) panel utilizes electrowetting technology to detect the most common causes of GP bacteremia (20 targets) and 4 antimicrobial resistance genes in positive blood culture bottles. Rapid detection of intrinsic vancomycin resistance and acquired resistance genes (mecA, mecC, vanA, vanB) enables early optimization of antimicrobial therapy whereas early detection of common contaminants decreases unnecessary antibiotic utilization and hospitalizations. Methods In this prospective study, we evaluated the performance of the BCID-GP panel compared to traditional standard of care culture and susceptibility testing with organism identification using the BioMerieux Vitek MS Matrix Assisted Laser Desorption Ionization (MALDI) Time of Flight mass spectrometry. Samples submitted for standard of care testing in Biomerieux BacT/Alert resin FA/FN blood culture bottles on the BacT/Alert VIRTUO automated blood culture system with GP bacteria on direct exam (n=100) were included. Results All GP bacteria were represented on the BCID-GP panel, most tests 97/100 (97%) yielded valid results, 53 common skin contaminants (50 coagulase negative staphylococci (CNS), 2 Bacillus, 1 Corynebacterium) were identified, and 7/7 coinfections with Gram negative (GN) bacteria were detected by the Pan GN target and identified by the BCID-GN panel. Discordant analyses revealed a positive percent agreement (PPA) of 96/97 (99%) with 1 false negative CNS and a negative percent agreement (NPA) of 92/97 (94.8%) with 5 false positives for either S. epidermidis or Corynebacterium. Detection of vanA yielded a PPA of 4/4 and NPA of 9/9. mecA gene detection exhibited a PPA of 14/14 and NPA of 14/14 for S. aureus and a PPA of 31/32 (97%) and NPA of 16/16 for coagulase negative staphylococci with 1 false negative methicillin resistant S. epidermidis. Conclusion Detection of acquired vancomycin resistance (n=4) and absence of mecA gene detection in Staphylococcus species (n=30) represent opportunities for early optimization of antimicrobial therapy in 34/100 (34%) of samples. The BCID-GP panel provides rapid accurate detection of resistant isolates and common contaminants enabling high quality data driven optimization of antimicrobial therapy. Disclosures Todd P. McCarty, MD, Cidara (Grant/Research Support)GenMark (Grant/Research Support, Other Financial or Material Support, Honoraria for Research Presentation)T2 Biosystems (Consultant) Sixto M. Leal, Jr., MD, PhD, Abnova (Grant/Research Support)AltImmune (Grant/Research Support)Amplyx Pharmaceuticals (Grant/Research Support)Astellas Pharmaceuticals (Grant/Research Support)CNINE Dx (Grant/Research Support)GenMark Diagnostics (Grant/Research Support, Other Financial or Material Support, Honoraria- Research Presentation)IHMA (Grant/Research Support)IMMY Dx (Grant/Research Support)JMI/Sentry (Grant/Research Support)mFluiDx Dx (Grant/Research Support)SpeeDx Dx (Grant/Research Support)Tetraphase Pharmaceuticals (Grant/Research Support)