Presence and Persistence of ESKAPEE Bacteria before and after Hospital Wastewater Treatment.

The metagenomic surveillance of antimicrobial resistance in wastewater has been suggested as a methodological tool to characterize the distribution, status, and trends of antibiotic-resistant bacteria. In this study, a cross-sectional collection of samples of hospital-associated raw and treated wastewater were obtained from February to March 2020. Shotgun metagenomic sequencing and bioinformatic analysis were performed to characterize bacterial abundance and antimicrobial resistance gene analysis. The main bacterial phyla found in all the samples were as follows: Proteobacteria, Bacteroides, Firmicutes, and Actinobacteria. At the species level, ESKAPEE bacteria such as E. coli relative abundance decreased between raw and treated wastewater, but S. aureus, A. baumannii, and P. aeruginosa increased, as did the persistence of K. pneumoniae in both raw and treated wastewater. A total of 172 different ARGs were detected; blaOXA, blaVEB, blaKPC, blaGES, mphE, mef, erm, msrE, AAC(6'), ant(3″), aadS, lnu, PBP-2, dfrA, vanA-G, tet, and sul were found at the highest abundance and persistence. This study demonstrates the ability of ESKAPEE bacteria to survive tertiary treatment processes of hospital wastewater, as well as the persistence of clinically important antimicrobial resistance genes that are spreading in the environment.

Ibero-Panamerican Federation of Periodontology Delphi study on the trends of periodontology and periodontics by the year 2030. A Latin American consensus.

BACKGROUND AND OBJECTIVE: As elsewhere in the world, the prevalence of periodontitis in stages I-II is high in the Latin American population, this scenario emphasizes the need for identification of urgent needs for allocating adequate resources to provide diagnosis, prevention, and treatment of these diseases. The aim of this Delphi study was to predict the trends in periodontology/periodontics in the Latin American region by the year 2030. METHODS: A steering committee and an advisory group of experts in periodontology/periodontics were selected from 16 countries. An open questionnaire of 60 questions was validated and used following the Delphi methodology. RESULTS: Two hundred and twenty-five experts from Latin America answered the two rounds of the questionnaire. Moderate to strong consensus was reached on 45 questions (75%). The prediction was that the prevalence of gingivitis and periodontitis in stages I and II will be maintained, the importance of the link with systemic diseases will increase, and the impact of prevention and periodontal treatment will also increase, mainly in the private sector. There was a strong consensus that plastic and regenerative surgical procedures will increase, as well as the demand for training in the specialty of periodontology. CONCLUSIONS: The present study has provided relevant and useful information on predictions in periodontology/periodontics in Latin America, with important level of consensus among experts. It has been predicted that periodontitis will still be a highly prevalent disease, and its links with other medical conditions should demand more attention by health authorities to develop adequate prevention and management policies and strategies.

Anxious to detect deceit: an empirical investigation of social defense theory.

Social Defense Theory (SDT) states that anxious attachment reflects an adaptive sentinel strategy, whereby anxious people should be better able to detect lies than secure people. Existing research on this issue, however, has not been able to evaluate whether heightened lie detection among anxious individuals is due to an actual ability or a bias to assume that others are lying (one that pays off when others are, in fact, lying). We addressed this issue in a study in which 254 adults had to determine whether people in videos were lying or telling the truth about their experiences. Contrary to the predictions of SDT, highly anxious people did not have a heightened ability to separate lies from truths, but were biased to assume that others were lying regardless of the authenticity of their statements.

Characterization of immunoglobulin loci in the gigantic genome of Ambystoma mexicanum.

Background: The axolotl, Ambystoma mexicanum is a unique biological model for complete tissue regeneration. Is a neotenic endangered species and is highly susceptible to environmental stress, including infectious disease. In contrast to other amphibians, the axolotl is particularly vulnerable to certain viral infections. Like other salamanders, the axolotl genome is one of the largest (32 Gb) and the impact of genome size on Ig loci architecture is unknown. To better understand the immune response in axolotl, we aimed to characterize the immunoglobulin loci of A. mexicanum and compare it with other model vertebrates. Methods: The most recently published genome sequence of A. mexicanum (V6) was used for alignment-based annotation and manual curation using previously described axolotl Ig sequences or reference sequences from other vertebrates. Gene models were further curated using A. mexicanum spleen RNA-seq data. Human, Xenopus tropicalis, Danio rerio (zebrafish), and eight tetrapod reference genomes were used for comparison. Results: Canonical A. mexicanum heavy chain (IGH), lambda (IGL), sigma (IGS), and the putative surrogate light chain (SLC) loci were identified. No kappa locus was found. More than half of the IGHV genes and the IGHF gene are pseudogenes and there is no clan I IGHV genes. Although the IGH locus size is proportional to genome size, we found local size restriction in the IGHM gene and the V gene intergenic distances. In addition, there were V genes with abnormally large V-intron sizes, which correlated with loss of gene functionality. Conclusion: The A. mexicanum immunoglobulin loci share the same general genome architecture as most studied tetrapods. Consistent with its large genome, Ig loci are larger; however, local size restrictions indicate evolutionary constraints likely to be imposed by high transcriptional demand of certain Ig genes, as well as the V(D)J recombination over very long genomic distance ranges. The A. mexicanum has undergone an extensive process of Ig gene loss which partially explains a reduced potential repertoire diversity that may contribute to its impaired antibody response.

Association of Gut Microbiota with Dietary-dependent Childhood Obesity.

AIM: To evaluate the taxonomic profile of the gut microbiota using metagenomics and the association with diet-dependent childhood obesity. METHODS: A cross-sectional study of a subsample of 46 children was conducted. The children were classified as normal-weight, overweight, and obese according to their age and sex and the World Health Organization (WHO) guidelines. Dietary patterns were determined through principal component analysis. The profile of the human gut microbiota was determined by bioinformatic analysis using whole metagenome shotgun sequencing. The association of gut microbiota and z-BMI, waist circumference and hip circumference, and the possible modifying effect of diet were analyzed using multiple regression models. RESULTS: Children with an abundance of Holdemania spp. and high protein and complex carbohydrate consumption had a lower z-BMI (ß -19.06, p = 0.011), waist circumference (ß -171.92, p = 0.003), and hip circumference (ß -157.57, p = 0.004). In contrast, observed a positive association between Coprococcus catus and the low intake of this dietary pattern with hip circumference (ß 147.87, p = 0.025). Furthermore, the presence of Bilophila spp. and Paraprevotella xylaniphila with high saturated fat and simple carbohydrate consumption we observed a positive association between z-BMI (ß 47.5, p = 0.002), hip circumference (ß 44.54, p = 0.025), and waist circumference (ß 44.34, p = 0.004). CONCLUSION: We suggest that the synergism between diet and the profile of children's gut microbiota can be a factor that could be associated with the development of obesity and its complications in childhood.

Put your mask on first to help others: Attachment and sentinel behavior during the COVID-19 pandemic.

The objective of this research was to learn whether attachment style is related to the ways people try to warn, protect, and care for others during the pandemic and what kinds, if any, personal protective measures they are taking. Data were collected in early May 2020 from 200 Amazon MTurk (AMT) workers who participated in exchange for payment. People who were high in attachment-related anxiety were more likely to behave as "sentinels" (i.e., warning loved ones to engage in safe practices such as hand washing, wearing a face mask), whereas those high in attachment avoidance were less likely to do so. These findings suggest that insecure attachment may contribute to peoples' willingness to protect themselves and others during the pandemic.

Author Correction: Analysis of sequencing strategies and tools for taxonomic annotation: Defining standards for progressive metagenomics.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Regeneración de tejido blando en una clase 3 mediante un colgajo desplazado lateral e injerto de tejido conjuntivo. Reporte de un caso clínico / Elian class 3 regeneration of soft tissue using a laterally moved flap and connective tissue graft. A clinical case report

RESUMEN: Se describe el caso clínico de una paciente de 60 años, sexo femenino, sana, la cual presentaba en el diente 1.1 una recesión de 6 mm de longitud con extensa pérdida ósea en vestibular correspondiente a una clase 3 de Elian. En el presente reporte se expone los pasos que se siguieron para llegar a una Clase 2 de Elian, mediante un colgajo desplazado lateral con injerto de tejido conjuntivo subepitelial.

ABSTRACT: We describe the clinical case of a 60-year-old female patient, healthy, who presented a 6mm-long recession on tooth 1.1 with extensive vestibular bone loss corresponding to an Elian class 3. In the present report, the steps followed to reach an Elian class 2,by means of a laterally moved flap with subepithelial connective tissue graft, are shown.

Estimates of donated sperm use in the United States: National Survey of Family Growth 1995-2017.

OBJECTIVE: To provide national estimates of donor insemination (DI) use in the United States and a description of the population of users. DESIGN: Population estimates were generated from nationally representative data through weighted proportions and count estimates. SETTING: Not applicable. PATIENT(S): Participants were U.S. women of childbearing age (15-44 years) sampled for interview in the National Survey of Family Growth. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Respondents who reported having received artificial insemination were asked the origin of the sperm. Responses could include husband/partner, donor only, or mixed donor and husband/partner. RESULT(S): In 1995, an estimated 170,701 (95% confidence interval 106,577-234,825) women had undergone DI using donor or mixed sperm. In 2015-2017, 440,986 (95% confidence interval 108,458-773,513) women were estimated to have used it. The DI users were mostly white, urban, older, college-educated, and had high family incomes. CONCLUSION(S): The DI use changed over time, from a decrease between 1995 and 2013 to a precipitous growth in 2015 to 2017. In recent years, nearly half a million women may be dealing with personal, relationship, and familial issues born of DI use. The United States does not maintain records on the usage of donor sperm, but better tracking of the use and outcomes of treatment would provide better estimates of the size of the affected population.

The effect of admitting fault versus shifting blame on expectations for others to do the same.

A wealth of research has investigated how and why people cast blame. However, less is known about blame-shifting (i.e., blaming someone else for one's own failures) and how exposure to a blame-shifting agent might lead to expectations that other agents will also shift blame. The present research tested whether exposure to a blame-shifting (versus responsibility-taking) agent would lead perceivers to expect a second, unrelated target to also shift blame. Contrary to our expectations, people expected greater blame-shifting after exposure to a responsible agent, particularly when perceivers were surprised by this reaction to failure. Discussion focuses on how people habitually expect some people to shift blame for their mishaps, and how expectancy violations when people act in unexpected ways predict the extent to which perceivers expect unrelated agents to also shift blame.

Analysis of sequencing strategies and tools for taxonomic annotation: Defining standards for progressive metagenomics.

Metagenomics research has recently thrived due to DNA sequencing technologies improvement, driving the emergence of new analysis tools and the growth of taxonomic databases. However, there is no all-purpose strategy that can guarantee the best result for a given project and there are several combinations of software, parameters and databases that can be tested. Therefore, we performed an impartial comparison, using statistical measures of classification for eight bioinformatic tools and four taxonomic databases, defining a benchmark framework to evaluate each tool in a standardized context. Using in silico simulated data for 16S rRNA amplicons and whole metagenome shotgun data, we compared the results from different software and database combinations to detect biases related to algorithms or database annotation. Using our benchmark framework, researchers can define cut-off values to evaluate the expected error rate and coverage for their results, regardless the score used by each software. A quick guide to select the best tool, all datasets and scripts to reproduce our results and benchmark any new method are available at https://github.com/Ales-ibt/Metagenomic-benchmark . Finally, we stress out the importance of gold standards, database curation and manual inspection of taxonomic profiling results, for a better and more accurate microbial diversity description.

Lower IgG somatic hypermutation rates during acute dengue virus infection is compatible with a germinal center-independent B cell response.

BACKGROUND: The study of human B cell response to dengue virus (DENV) infection is critical to understand serotype-specific protection and the cross-reactive sub-neutralizing response. Whereas the first is beneficial and thus represents the ultimate goal of vaccination, the latter has been implicated in the development of severe disease, which occurs in a small, albeit significant, fraction of secondary DENV infections. Both primary and secondary infections are associated with the production of poly-reactive and cross-reactive IgG antibodies. METHODS: To gain insight into the effect of DENV infection on the B cell repertoire, we used VH region high-throughput cDNA sequencing of the peripheral blood IgG B cell compartment of 19 individuals during the acute phase of infection. For 11 individuals, a second sample obtained 6 months later was analyzed for comparison. Probabilities of sequencing antibody secreting cells or memory B cells were estimated using second-order Monte Carlo simulation. RESULTS: We found that in acute disease there is an increase in IgG B cell diversity and changes in the relative use of segments IGHV1-2, IGHV1-18, and IGHV1-69. Somewhat unexpectedly, an overall low proportion of somatic hypermutated antibody genes was observed during the acute phase plasmablasts, particularly in secondary infections and those cases with more severe disease. CONCLUSIONS: Our data are consistent with an innate-like antiviral recognition system mediated by B cells using defined germ-line coded B cell receptors, which could provide a rapid germinal center-independent antibody response during the early phase of infection. A model describing concurrent T-dependent and T-independent B cell responses in the context of DENV infection is proposed, which incorporates the selection of B cells using hypomutated IGHV segments and their potential role in poly/cross-reactivity. Its formal demonstration could lead to a definition of its potential implication in antibody-dependent enhancement, and may contribute to rational vaccine development efforts.

Longitudinal analysis of the peripheral B cell repertoire reveals unique effects of immunization with a new influenza virus strain.

BACKGROUND: Despite the potential to produce antibodies that can neutralize different virus (heterotypic neutralization), there is no knowledge of why vaccination against influenza induces protection predominantly against the utilized viral strains (homotypic response). Identification of structural patterns of the B cell repertoire associated to heterotypic neutralization may contribute to identify relevant epitopes for a universal vaccine against influenza. METHODS: Blood samples were collected from volunteers immunized with 2008/2009 trivalent inactivated vaccine (TIV), pandemic H1N1 (pdmH1N1) monovalent inactivated vaccine (MIV) and the 2014/2015 TIV. Neutralization was assessed by hemagglutination and microneutralization test. IgG V(H) amplicons derived from peripheral blood RNA from pre-immune and 7 days post vaccination were subjected to 454-Roche sequencing. Full reconstruction of the sampled repertoires was done with ImmunediveRsity. RESULTS: The TIV induced a predominantly homotypic neutralizing serologic response, while the 09 MIV induced a heterotypic neutralizing seroconversion in 17% of the individuals. Both the 08/09 and the 14/15 TIV were associated with a reduction in clonotypic diversity, whereas 09 MIV was the opposite. Moreover, TIV and MIV induced distinctive patterns of IGHV segment use that are consistent with B cell selection by conserved antigenic determinants shared by the pre-pandemic and the pandemic strains. However, low somatic hypermutation rates in IgG after 09 MIV immunization, but not after 08/09 and 14/15 TIV immunization were observed. Furthermore, no evidence of the original antigenic sin was found in the same individuals after vaccination with the three vaccines. CONCLUSIONS: Immunization with a new influenza virus strain (2009 pdmH1N1) induced unique effects in the peripheral B cell repertoire clonal structure, a stereotyped response involving distinctive IGHV segment use and low somatic hypermutation levels. These parameters were contrastingly different to those observed in response to pre-pandemic and post-pandemic vaccination, and may be the result of clonal selection of common antigenic determinants, as well as germinal center-independent responses that wane as the pandemic strain becomes seasonal. Our findings may contribute in the understanding of the structural and cellular basis required to develop a universal influenza vaccine.

Reconstructing and mining the B cell repertoire with ImmunediveRsity.

The B cell antigen receptor repertoire is highly diverse and constantly modified by clonal selection. High-throughput DNA sequencing (HTS) of the lymphocyte repertoire (Rep-Seq) represents a promising technology to explore such diversity ex-vivo and assist in the identification of antigen-specific antibodies based on molecular signatures of clonal selection. Therefore, integrative tools for repertoire reconstruction and analysis from antibody sequences are needed. We developed ImmunediveRity, a stand-alone pipeline primarily based in R programming for the integral analysis of B cell repertoire data generated by HTS. The pipeline integrates GNU software and in house scripts to perform quality filtering, sequencing noise correction and repertoire reconstruction based on V, D and J segment assignment, clonal origin and unique heavy chain identification. Post-analysis scripts generate a wealth of repertoire metrics that in conjunction with a rich graphical output facilitates sample comparison and repertoire mining. Its performance was tested with raw and curated human and mouse 454-Roche sequencing benchmarks providing good approximations of repertoire structure. Furthermore, ImmunediveRsity was used to mine the B cell repertoire of immunized mice with a model antigen, allowing the identification of previously validated antigen-specific antibodies, and revealing different and unexpected clonal diversity patterns in the post-immunization IgM and IgG compartments. Although ImmunediveRsity is similar to other recently developed tools, it offers significant advantages that facilitate repertoire analysis and repertoire mining. ImmunediveRsity is open source and free for academic purposes and it runs on 64 bit GNU/Linux and MacOS. Available at: https://bitbucket.org/ImmunediveRsity/immunediversity/.

Cirugía plástica periodontal, una realidad asistencial: reporte de casos / Periodontal plastic surgery, a care reality: case report

En la actualidad, la periodoncia no solo se encarga de resolver los procesos infecciosos que afectan al periodonto, sino que también es responsable de la preservación de la función, del confort y de la estética de los tejidos periodontales. Es por eso que hoy en día las técnicas de cirugía plástica periodontal son una herramienta esencial para la resolución de defectos mucogingivales en donde el tejido conectivo subepitelial, debido a su biología, se ha convertido en la mejor alternativa para resolver las secuelas derivadas de trauma maxilofacial, tratamientos de ortodoncia en biotipos finos, trauma dentoalveolar, resección de tumores, tratamiento de las secuelas de la periodontitis, en donde al restaurar la pérdida de tejido existente y modificar el biotipo periodontal mejora el pronóstico dentario. En Chile, el tratamiento periodontal realizado en la atención secundaria se ha centrado históricamente en el control de infecciones. Sin embargo, debido a la gran demanda de casos relacionados con los tejidos periodontales sin periodontitis, hoy enfrentamos una nueva realidad clínica asistencial que amerita el empleo de nuevas técnicas y procedimientos quirúrgicos tendentes a la restauración de los tejidos periodontales perdidos o mutilados. En este reporte de casos se expone la labor realizada en el servicio dental del Hospital El Salvador, dependiente del Servicio de Salud Metropolitano Oriente, en donde desde el año 2006 se realizan intervenciones quirúrgicas con técnicas de cirugía plástica periodontal para resolver casos complejos.

Periodontics is not only charged with resolving infectious processes that affect the periodontium, but is also responsible for preservation of function, comfort and aesthetics of the periodontal tissues. For this reason current plastic periodontal surgery techniques are an essential tool for the resolution of mucogingival defects, where the subepithelial connective tissues, due to their biology, have become the best alternative to resolve the sequelae of maxillo-facial trauma (TMF), fine biotype orthodontic treatment, dentoalveolar trauma (TDA), tumor resection, and treatment of the sequelae of periodontitis. In all of these conditions, the restoration of lost tissue and the modification of existing periodontal biotype dental improves their prognosis. In Chile, the periodontal treatment performed in secondary care has historically focused on infection control. However, due to the high demand for cases involving periodontal tissues without periodontitis, there is now a new reality that must focus clinical attention on the use of new techniques and surgical procedures designed to restore lost or mutilated periodontal tissues. This case report presents the work of the El Salvador Hospital Dental Department of East Metropolitan Health Service (SSMO), where periodontal plastic surgery techniques to treat complex cases have been performed since 2006.

Hybridization-based antibody cDNA recovery for the production of recombinant antibodies identified by repertoire sequencing.

High-throughput sequencing of the antibody repertoire is enabling a thorough analysis of B cell diversity and clonal selection, which may improve the novel antibody discovery process. Theoretically, an adequate bioinformatic analysis could allow identification of candidate antigen-specific antibodies, requiring their recombinant production for experimental validation of their specificity. Gene synthesis is commonly used for the generation of recombinant antibodies identified in silico. Novel strategies that bypass gene synthesis could offer more accessible antibody identification and validation alternatives. We developed a hybridization-based recovery strategy that targets the complementarity-determining region 3 (CDRH3) for the enrichment of cDNA of candidate antigen-specific antibody sequences. Ten clonal groups of interest were identified through bioinformatic analysis of the heavy chain antibody repertoire of mice immunized with hen egg white lysozyme (HEL). cDNA from eight of the targeted clonal groups was recovered efficiently, leading to the generation of recombinant antibodies. One representative heavy chain sequence from each clonal group recovered was paired with previously reported anti-HEL light chains to generate full antibodies, later tested for HEL-binding capacity. The recovery process proposed represents a simple and scalable molecular strategy that could enhance antibody identification and specificity assessment, enabling a more cost-efficient generation of recombinant antibodies.

Pandemic influenza A(H1N1) 2009 and respiratory syncytial virus associated hospitalizations.

OBJECTIVES: To determine the contribution of influenza and respiratory syncytial virus (RSV) as the cause of lower respiratory tract infection (LRTI) associated hospitalizations during the first year of the influenza A(H1N1) 2009 pandemic and to assess the severity of illness during the second pandemic wave. METHODS: Patients admitted with LRTI from April 2009 through March 2010 were assessed for the presence of influenza and RSV. Pandemic influenza virus was detected by means of a nested RT-PCR assay and/or the CDC's real time-PCR protocol. RSV was detected using a one-step RT-PCR assay. The characteristics of patients admitted during the first and second pandemic outbreaks were compared. RESULTS: 657 patients with LRTI were admitted during the study period. Pandemic influenza virus was detected in 180 and RSV in 133. Influenza was the most common cause of infection in adults, while RSV was more common in children. There were no differences in disease severity between the first and second pandemic outbreaks. CONCLUSIONS: Pandemic influenza virus was associated to increased numbers of hospitalizations and deaths; particularly in adults. The severity of the first and second pandemic outbreaks was similar. RSV continues to be the main pathogen responsible for hospitalizations in young children.

Evaluación de la presión arterial y la función renal en los habitantes mayores de 14 años del poblado rural de Huallín- Ancash / Evaluation of the arterial pressure and the renal function in the greater in habitants of 14 years of the rural town of Huallin-Ancash

Objetivo: Hacer un despistaje de hipertensión arterial (HTA) y enfermedad renal en una población rural de altura. Métodos: Se estudió a los habitantes del poblado rural de Huallin (Ancash- Perú) ubicado a 3,00 metros sobre el nivel del mar, quienes se midió la presión arterial (en tres oportunidades), se definió HTA si la PA> 140/90, se midió la proteinuria y hematuria con tiras reactivas. Los resultados fueron evaluados con el programa estadístico Epinfo. Resultados: Se evaluó a un total de 201 pobladores mayores de 14 años. La frecuencia de HTA sistólica aislada fue del 2 por ciento, la HTA diástolica fue de 23 por ciento, siendo más frecuente en el sexo masculino (31 por ciento vs 18 por ciento, p<0.05). La HTA diastólica en estado I fue las más frecuente en ambos sexos (70.6 por ciento). La frecuencia de proteinuria fue de 5 por ciento y la de hematuria fue de 13 por ciento. La PA sistólica fue mayor en los pacientes con proteinuria (127 vs 115 mmHg, p=0.02). Conclusión: La frecuencia de HTA y proteinuria, es superior a los reportado previamente para poblaciones de altura, la PA sistólica fue mayor en los pobladores con proteinuria, estos resultados sugieren un cambio en el perfil epidemiológico de la población rural de altura.