Single-cell dissection of the human motor and prefrontal cortices in ALS and FTLD.

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) share many clinical, pathological, and genetic features, but a detailed understanding of their associated transcriptional alterations across vulnerable cortical cell types is lacking. Here, we report a high-resolution, comparative single-cell molecular atlas of the human primary motor and dorsolateral prefrontal cortices and their transcriptional alterations in sporadic and familial ALS and FTLD. By integrating transcriptional and genetic information, we identify known and previously unidentified vulnerable populations in cortical layer 5 and show that ALS- and FTLD-implicated motor and spindle neurons possess a virtually indistinguishable molecular identity. We implicate potential disease mechanisms affecting these cell types as well as non-neuronal drivers of pathogenesis. Finally, we show that neuron loss in cortical layer 5 tracks more closely with transcriptional identity rather than cellular morphology and extends beyond previously reported vulnerable cell types.

Transcription factor NAC1 activates expression of peptidase-encoding AtCEPs in roots to limit root hair growth.

Plant genomes encode a unique group of papain-type Cysteine EndoPeptidases (CysEPs) containing a KDEL endoplasmic reticulum (ER) retention signal (KDEL-CysEPs or CEPs). CEPs process the cell-wall scaffolding EXTENSIN (EXT) proteins that regulate de novo cell-wall formation and cell expansion. Since CEPs cleave EXTs and EXT-related proteins, acting as cell-wall-weakening agents, they may play a role in cell elongation. The Arabidopsis (Arabidopsis thaliana) genome encodes 3 CEPs (AtCPE1-AtCEP3). Here, we report that the genes encoding these 3 Arabidopsis CEPs are highly expressed in root-hair (RH) cell files. Single mutants have no evident abnormal RH phenotype, but atcep1-3 atcep3-2 and atcep1-3 atcep2-2 double mutants have longer RHs than wild-type (Wt) plants, suggesting that expression of AtCEPs in root trichoblasts restrains polar elongation of the RH. We provide evidence that the transcription factor NAC1 (petunia NAM and Arabidopsis ATAF1, ATAF2, and CUC2) activates AtCEPs expression in roots to limit RH growth. Chromatin immunoprecipitation indicates that NAC1 binds to the promoter of AtCEP1, AtCEP2, and, to a lower extent, AtCEP3 and may directly regulate their expression. Inducible NAC1 overexpression increases AtCEP1 and AtCEP2 transcript levels in roots and leads to reduced RH growth while the loss of function nac1-2 mutation reduces AtCEP1-AtCEP3 gene expression and enhances RH growth. Likewise, expression of a dominant chimeric NAC1-SRDX repressor construct leads to increased RH length. Finally, we show that RH cell walls in the atcep1-3 atcep3-2 double mutant have reduced levels of EXT deposition, suggesting that the defects in RH elongation are linked to alterations in EXT processing and accumulation. Our results support the involvement of AtCEPs in controlling RH polar growth through EXT processing and insolubilization at the cell wall.

Exploring morphological variation in bismuth ferrite nanostructures by pulsed laser deposition: synthesis, structural and electrochemical properties.

Structural and electrochemical properties of bismuth ferrite nanostructures produced by pulsed laser deposition with various morphologies are reported. The nanostructures are also explored as electrode materials for high-performance supercapacitors. Scanning electron microscopy images revealed that various bismuth ferrite morphologies were produced by varying the background pressure (10-6, 0.01, 0.10, 0.25, 0.50, 1.0, 2.0 and 4.0 Torr) in the deposition chamber and submitting them to a thermal treatment after deposition at 500◦C. The as-deposited bismuth ferrite nanostructures range from very compact thin-film (10-6, 0.01, 0.10 Torr), to clustered nanoparticles (0.25, 0.50, 1.0 Torr), to very dispersed arrangement of nanoparticles (2.0 and 4.0 Torr). The electrochemical characteristic of the electrodes was investigated through cyclic voltammetry process. The increase in the specific surface area of the nanostructures as background pressure in the chamber increases does not lead to an increase in interfacial capacitance. This is likely due to the wakening of electrical contact between nanoparticles with increasing porosity of the nanostructures. The thermal treatment increased the contact between nanoparticles, which caused an increase in the interfacial capacitance of the nanostructure deposited under high background pressure in the chamber.

Randomized controlled trials in nursing conducted by Latin American research teams: A scoping review.

INTRODUCTION: Randomized controlled trials (RCTs) are the cornerstone of systematic reviews and other evidence synthesis. RCT identification remains challenging because of limitations in their indexation in major databases and potential language bias. Scientific production in Latin American nursing is steadily increasing, but little is known about its design or main features. We aimed to identify the extent of evidence from RCTs in nursing conducted by Latin American research teams and evaluate their main characteristics, including potential risk of bias. DESIGN: Scoping review with risk of bias assessment. METHODS: We conducted a scoping review including a comprehensive electronic search in five relevant databases. We completed a descriptive data analysis and a risk of bias assessment of eligible studies using Cochrane's guidance. RESULTS: We identified 1784 references of which 47 were RCTs published in 40 journals. Twenty (42.6%) RCTs were published in journals in English. Chronic diseases were the most common health conditions studied (29.7%). Fifteen (31.9%) RCTs had a high risk of bias. Thirty (75%) journals were included in the Journal Citation Report (JCR) catalog and 5 (16.7%) were journals classified under nursing category. Twenty-one (52.5%) journals explicitly required CONSORT checklist recommendations for RCTs reporting. CONCLUSION: Publication of RCTs in nursing by Latin American authors has increased. Most journals where RCTs are published are in English and not specific to nursing. Searches in journals of other disciplines may be necessary to facilitate identification of RCTs in nursing. CONSORT statements need to be actively promoted to facilitate rigorous methodology and reporting of RCTs. CLINICAL RELEVANCE STATEMENT: This study highlights the need for an increased research focus on RCTs in nursing in Latin America, and the importance of enhancing the reporting quality of these studies to support evidence-based nursing practice.

A Bioorthogonal Dual Fluorogenic Probe for the Live-Cell Monitoring of Nutrient Uptake by Mammalian Cells.

Cells rely heavily on the uptake of exogenous nutrients for survival, growth, and differentiation. Yet quantifying the uptake of small molecule nutrients at the single cell level is difficult. Here we present a new approach to studying the nutrient uptake in live single cells using Inverse Electron-Demand Diels Alder (IEDDA) chemistry. We have modified carboxyfluorescein-diacetate-succinimidyl esters (CFSE)-a quenched fluorophore that can covalently react with proteins and is only turned on in the cytosol of a cell following esterase activity-with a tetrazine. This tetrazine serves as a second quencher for the pendant fluorophore. Upon reaction with nutrients modified with an electron-rich or strained dienophile in an IEDDA reaction, this quenching group is destroyed, thereby enabling the probe to fluoresce. This has allowed us to monitor the uptake of a variety of dienophile-containing nutrients in live primary immune cell populations using flow cytometry and live-cell microscopy.

A Lysosome-Targeted Tetrazine for Organelle-Specific Click-to-Release Chemistry in Antigen Presenting Cells.

Bioorthogonal deprotections are readily used to control biological function in a cell-specific manner. To further improve the spatial resolution of these reactions, we here present a lysosome-targeted tetrazine for an organelle-specific deprotection reaction. We show that trans-cyclooctene deprotection with this reagent can be used to control the biological activity of ligands for invariant natural killer T cells in the lysosome to shed light on the processing pathway in antigen presenting cells. We then use the lysosome-targeted tetrazine to show that long peptide antigens used for CD8+ T cell activation do not pass through this organelle, suggesting a role for the earlier endosomal compartments for their processing.

GK-1 effectively reduces angiogenesis and prevents T cell exhaustion in a breast cancer murine experimental model.

Breast cancer is the leading malignancy in women worldwide, both in terms of incidence and mortality. Triple-negative breast cancer (TNBC) is the type with the worst clinical outcomes and with fewer therapeutic options than other types of breast cancer. GK-1 is a peptide that in the experimental model of the metastatic 4T1 breast cancer has demonstrated anti-tumor and anti-metastatic properties. Herein, GK-1 (5 mg/kg, i.v.) weekly administrated not only decreases tumor growth and the number of lung macro-metastases but also lung and lymph nodes micro-metastases. Histological analysis reveals that GK-1 reduced 57% of the intra-tumor vascular areas, diminished the leukemoid reaction's progression, and the spleens' weight and length. A significant reduction in VEGF-C, SDF-1, angiopoietin-2, and endothelin-1 angiogenic factors was induced. Moreover, GK-1 prevents T cell exhaustion in the tumor-infiltrating lymphocytes (TILs) decreasing PD-1 expression. It also increased IFN-γ and granzyme-B expression and the cytotoxic activity of CD8+ TILs cells against tumor cells. All these features were found to be associated with a better antitumor response and prognosis. Altogether, these results reinforce the potential of GK-1 to improve the clinical outcome of triple-negative breast cancer immunotherapy. Translation research is ongoing towards its evaluation in humans.

Deciphering a shared transcriptomic regulation and the relative contribution of each regulator type through endometrial gene expression signatures.

BACKGORUND: While various endometrial biomarkers have been characterized at the transcriptomic and functional level, there is generally a poor overlap among studies, making it unclear to what extent their upstream regulators (e.g., ovarian hormones, transcription factors (TFs) and microRNAs (miRNAs)) realistically contribute to menstrual cycle progression and function. Unmasking the intricacies of the molecular interactions in the endometrium from a novel systemic point of view will help gain a more accurate perspective of endometrial regulation and a better explanation the molecular etiology of endometrial-factor infertility. METHODS: An in-silico analysis was carried out to identify which regulators consistently target the gene biomarkers proposed in studies related to endometrial progression and implantation failure (19 gene lists/signatures were included). The roles of these regulators, and of genes related to progesterone and estrogens, were then analysed in transcriptomic datasets compiled from samples collected throughout the menstrual cycle (n = 129), and the expression of selected TFs were prospectively validated in an independent cohort of healthy participants (n = 19). RESULTS: A total of 3,608 distinct genes from the 19 gene lists were associated with endometrial progression and implantation failure. The lists' regulation was significantly favoured by TFs (89% (17/19) of gene lists) and progesterone (47% (8 /19) of gene lists), rather than miRNAs (5% (1/19) of gene lists) or estrogen (0% (0/19) of gene lists), respectively (FDR < 0.05). Exceptionally, two gene lists that were previously associated with implantation failure and unexplained infertility were less hormone-dependent, but primarily regulated by estrogen. Although endometrial progression genes were mainly targeted by hormones rather than non-hormonal contributors (odds ratio = 91.94, FDR < 0.05), we identified 311 TFs and 595 miRNAs not previously associated with ovarian hormones. We highlight CTCF, GATA6, hsa-miR-15a-5p, hsa-miR-218-5p, hsa-miR-107, hsa-miR-103a-3p, and hsa-miR-128-3p, as overlapping novel master regulators of endometrial function. The gene expression changes of selected regulators throughout the menstrual cycle (FDR < 0.05), dually validated in-silico and through endometrial biopsies, corroborated their potential regulatory roles in the endometrium. CONCLUSIONS: This study revealed novel hormonal and non-hormonal regulators and their relative contributions to endometrial progression and pathology, providing new leads for the potential causes of endometrial-factor infertility.

Testing the hierarchy of predictability in grassland restoration across a gradient of environmental severity.

Ecological restoration is critical for recovering degraded ecosystems but is challenged by variable success and low predictability. Understanding which outcomes are more predictable and less variable following restoration can improve restoration effectiveness. Recent theory asserts that the predictability of outcomes would follow an order from most to least predictable from coarse to fine community properties (physical structure > taxonomic diversity > functional composition > taxonomic composition) and that predictability would increase with more severe environmental conditions constraining species establishment. We tested this "hierarchy of predictability" hypothesis by synthesizing outcomes along an aridity gradient with 11 grassland restoration projects across the United States. We used 1829 vegetation monitoring plots from 227 restoration treatments, spread across 52 sites. We fit generalized linear mixed-effects models to predict six indicators of restoration outcomes as a function of restoration characteristics (i.e., seed mixes, disturbance, management actions, time since restoration) and used variance explained by models and model residuals as proxies for restoration predictability. We did not find consistent support for our hypotheses. Physical structure was among the most predictable outcomes when the response variable was relative abundance of grasses, but unpredictable for total canopy cover. Similarly, one dimension of taxonomic composition related to species identities was unpredictable, but another dimension of taxonomic composition indicating whether exotic or native species dominated the community was highly predictable. Taxonomic diversity (i.e., species richness) and functional composition (i.e., mean trait values) were intermittently predictable. Predictability also did not increase consistently with aridity. The dimension of taxonomic composition related to the identity of species in restored communities was more predictable (i.e., smaller residuals) in more arid sites, but functional composition was less predictable (i.e., larger residuals), and other outcomes showed no significant trend. Restoration outcomes were most predictable when they related to variation in dominant species, while those responding to rare species were harder to predict, indicating a potential role of scale in restoration predictability. Overall, our results highlight additional factors that might influence restoration predictability and add support to the importance of continuous monitoring and active management beyond one-time seed addition for successful grassland restoration in the United States.

Vulvovaginal yeast infections during pregnancy and perinatal outcomes: systematic review and meta-analysis.

BACKGROUND: Vulvovaginal yeast infections in pregnancy are common and can cause extensive inflammation, which could contribute to adverse pregnancy outcomes. Symptomatic yeast infections are likely to cause more inflammation than asymptomatic. The objective of this study was to investigate associations between symptomatic and asymptomatic vulvovaginal yeast infections in pregnancy and perinatal outcomes. METHODS: We did a systematic review and searched eight databases until 01 July 2022. We included studies reporting on pregnant women with and without laboratory confirmed vulvovaginal yeast infection and preterm birth or eight other perinatal outcomes. We used random effects meta-analysis to calculate summary odds ratios (OR), 95% confidence intervals (CI) and prediction intervals for the association between yeast infection and outcomes. We described findings from studies with multivariable analyses. We assessed the risk of bias using published tools. RESULTS: We screened 3909 references and included 57 studies. Only 22/57 studies reported information about participant vulvovaginal symptoms. Preterm birth was an outcome in 35/57 studies (49,161 women). In 32/35 studies with available data, the summary OR from univariable analyses was 1.01 (95% CI 0.84-1.21, I2 60%, prediction interval 0.45-2.23). In analyses stratified by symptom status, we found ORs of 1.44 (95% CI 0.92-2.26) in two studies with ≥ 50% symptomatic participants, 0.84 (95% CI 0.45-1.58) in seven studies with < 50% symptomatic participants, and 1.12 (95% CI 0.94-1.35) in four studies with asymptomatic participants. In three studies with multivariable analysis, adjusted ORs were greater than one but CIs were compatible with there being no association. We did not find associations between vulvovaginal yeast infection and any secondary outcome. Most studies were at high risk of bias in at least one domain and only three studies controlled for confounding. CONCLUSIONS: We did not find strong statistical evidence of an increased risk for preterm birth or eight other adverse perinatal outcomes, in pregnant women with either symptomatic or asymptomatic vulvovaginal yeast infection. The available evidence is insufficient to make recommendations about testing and treatment of vulvovaginal yeast infection in pregnancy. Future studies should assess vulvovaginal symptoms, yeast organism loads, concomitant vaginal or cervical infections, and microbiota using state-of-the-art diagnostics. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42020197564.

Oral cancers preceded by proliferative verrucous leukoplakia exhibit distinctive molecular features.

OBJECTIVE: Proliferative verrucous leukoplakia (PVL) has high rates of malignant transformation into oral squamous cell carcinoma (OSCC), but the clinical and evolutionary pattern of OSCC from PVL (PVL-OSCC) is more favorable than that of OSCC not preceded by PVL (OSCC). Here, we aimed to explore the pathophysiologic differences between PVL-OSCC and OSCC through transcriptomic and DNA methylation analyses. MATERIALS AND METHODS: In this case-control study, oral biopsies from 8 PVL-OSCC and 10 OSCC patients were obtained for global sequencing using RNAseq and a genome-wide DNA methylation analysis via the Infinium EPIC Platform (graphical abstract). RESULTS: One hundred and thirty-three differentially expressed genes (DEGs) were detected, 94 of them upregulated in OSCC. Most of these genes were previously described in cancer and associated with prognosis. The integrative analysis revealed 26 DEGs, corresponding to 37 CpGs, whose promoters were regulated by DNA methylation. Twenty-nine of the CpGs were found as hypermethylated in PVL-OSCC. Only 5 of the genes that were aberrantly methylated and differentially expressed were upregulated in PVL-OSCC patients, whereas 21 were underexpressed. CONCLUSIONS: PVL-OSCC patients presented lower expression of cancer-related genes. Hypermethylation of the promoter region of many genes was also noticed, indicating that DNA methylation could be a regulatory mechanism.

Condom use and HIV testing among adults in Switzerland: repeated national cross-sectional surveys 2007, 2012, and 2017.

BACKGROUND: Monitoring of HIV and sexually transmitted infection (STI) prevention is important for guiding national sexual health programmes for both the general population and key populations. The objectives of this study were to examine trends and patterns of condom use at last intercourse and lifetime HIV testing in 2007, 2012 and 2017 in Switzerland, and to explore factors associated with these behaviours in men and women with opposite-sex partners and with same sex partners. METHODS: We analysed data from the 2007, 2012 and 2017 Swiss Health Survey. For each time point, outcome and population group, we conducted a descriptive analysis of weighted data and conducted multivariable logistic regression to obtain adjusted odds ratios (aOR) with 95% confidence intervals (CI) and compared outcomes between the timepoints. RESULTS: In total, 46,320 people were interviewed: 21,847 men and 23,141 women, who reported having sex only with partners of the opposite sex, 633 men who reported sex with a male partner and 699 women who reported sex with a female partner. Among the three surveys the prevalence of condom did not change but varied from 22 to 26% of men and 15 to 21% in women with only opposite-sex partners (aOR men, 0.93, 95% CI 0.82, 1.06; women 0.98, 95% CI 0.86 to 1.11). In men with any same sex partner the prevalence of condom use was 40% in 2007, 33% in 2012 and 54% in 2017 (aOR 1.80, 95% CI 0.97, 3.34). In multivariable analysis, the factor most strongly associated with condom use was sex with an occasional partner at last intercourse. HIV testing ever increased across all three survey years in people with opposite sex partners: 2017 vs. 2007, aOR men with only opposite-sex partners 1.64 (95% CI 1.49, 1.82), women with only opposite-sex partners 1.67 (1.51, 1.85), men with any same sex partner 0.98 (0.49, 1.96), women with any same sex partner 1.31 (0.74, 2.30). CONCLUSIONS: Monitoring of condom use, and HIV testing should continue and contribute to the development of the national sexual health programme. Stronger promotion of condoms for people with opposite-sex partners might be needed, since overall condom use at last intercourse has not changed since 2007.

Application of Natural Language Processing in Total Joint Arthroplasty: Opportunities and Challenges.

Total joint arthroplasty is becoming one of the most common surgeries within the United States, creating an abundance of analyzable data to improve patient experience and outcomes. Unfortunately, a large majority of this data is concealed in electronic health records only accessible by manual extraction, which takes extensive time and resources. Natural language processing (NLP), a field within artificial intelligence, may offer a viable alternative to manual extraction. Using NLP, a researcher can analyze written and spoken data and extract data in an organized manner suitable for future research and clinical use. This article will first discuss common subtasks involved in an NLP pipeline, including data preparation, modeling, analysis, and external validation, followed by examples of NLP projects. Challenges and limitations of NLP will be discussed, closing with future directions of NLP projects, including large language models.

An Overview of Machine Learning in Orthopedic Surgery: An Educational Paper.

The growth of artificial intelligence combined with the collection and storage of large amounts of data in the electronic medical record collection has created an opportunity for orthopedic research and translation into the clinical environment. Machine learning (ML) is a type of artificial intelligence tool well suited for processing the large amount of available data. Specific areas of ML frequently used by orthopedic surgeons performing total joint arthroplasty include tabular data analysis (spreadsheets), medical imaging processing, and natural language processing (extracting concepts from text). Previous studies have discussed models able to identify fractures in radiographs, identify implant type in radiographs, and determine the stage of osteoarthritis based on walking analysis. Despite the growing popularity of ML, there are limitations including its reliance on "good" data, potential for overfitting, long life cycle for creation, and ability to only perform one narrow task. This educational article will further discuss a general overview of ML, discussing these challenges and including examples of successfully published models.

Educational Overview of the Concept and Application of Computer Vision in Arthroplasty.

Image data has grown exponentially as systems have increased their ability to collect and store it. Unfortunately, there are limits to human resources both in time and knowledge to fully interpret and manage that data. Computer Vision (CV) has grown in popularity as a discipline for better understanding visual data. Computer Vision has become a powerful tool for imaging analytics in orthopedic surgery, allowing computers to evaluate large volumes of image data with greater nuance than previously possible. Nevertheless, even with the growing number of uses in medicine, literature on the fundamentals of CV and its implementation is mainly oriented toward computer scientists rather than clinicians, rendering CV unapproachable for most orthopedic surgeons as a tool for clinical practice and research. The purpose of this article is to summarize and review the fundamental concepts of CV application for the orthopedic surgeon and musculoskeletal researcher.

Biological Use of Nanostructured Silica-Based Materials Functionalized with Metallodrugs: The Spanish Perspective.

Since the pioneering work of Vallet-Regí's group on the design and synthesis of mesoporous silica-based materials with therapeutic applications, during the last 15 years, the potential use of mesoporous silica nanostructured materials as drug delivery vehicles has been extensively explored. The versatility of these materials allows the design of a wide variety of platforms that can incorporate numerous agents of interest (fluorophores, proteins, drugs, etc.) in a single scaffold. However, the use of these systems loaded with metallodrugs as cytotoxic agents against different diseases and with distinct therapeutic targets has been studied to a much lesser extent. This review will focus on the work carried out in this field, highlighting both the pioneering and recent contributions of Spanish groups that have synthesized a wide variety of systems based on titanium, tin, ruthenium, copper and silver complexes supported onto nanostructured silica. In addition, this article will also discuss the importance of the structural features of the systems for evaluating and modulating their therapeutic properties. Finally, the most interesting results obtained in the study of the potential therapeutic application of these metallodrug-functionalized silica-based materials against cancer and bacteria will be described, paying special attention to preclinical trials in vivo.

New Carbonate-Based Materials and Study of Cytotoxic Capacity in Cancer Cells.

Calcium carbonate, one of the most commonly found biominerals produced by organisms, has shown great potential for the development of systems with biological applications due to its excellent biocompatibility, biodegradability, and simple chemical composition. Here, we focus on the synthesis of various carbonate-based materials with vaterite phase control and their subsequent functionalization for applications in treating glioblastoma, one of the most limiting tumors currently without effective treatments. The incorporation of l-cysteine into the systems increased cell selectivity while the incorporation of manganese supplied the materials with cytotoxic capacity. Extensive characterization of the systems by infrared spectroscopy, ultraviolet-visible spectroscopy, X-ray diffraction, X-ray fluorescence, and transmission electron microscopy confirmed the incorporation of the different fragments causing selectivity and cytotoxicity to the systems. To verify their therapeutic activity, the vaterite-based materials were tested in the CT2A cell line (murine glioma) and compared to SKBR3 (breast cancer) and HEK-293T (human kidney) cell lines. These studies on the cytotoxicity of the materials have shown promising results that can encourage future in vivo studies in glioblastoma models.

Evaluation of the sealing capability and morphological fit of prefabricated dowels and fiber-reinforced composite resin: An in vitro study.

STATEMENT OF PROBLEM: Prefabricated dowels do not always provide intraradicular sealing in the root canal dentin, and the lack of sealing predisposes the dowel to adhesive failure and debonding. It is unclear if fiber-reinforced composite resin dowels provide better sealing. PURPOSE: The purpose of this in vitro study was to evaluate the intraradicular sealing and morphological fit of prefabricated dowels and fiber-reinforced composite resin dowels in root canal dentin. The thickness of the resin cement layer and push-out bond strength were determined to assess their effects on the sealing of the dowels. MATERIAL AND METHODS: A total of 50 permanent maxillary central incisors were endodontically treated and divided into 2 subgroups (n=25). In one group, prefabricated dowels were cemented; in the second group, fiber-reinforced composite resin dowels were placed. The thickness of the resin cement layer was assessed in 3 different locations: coronal, middle, and apical of the root canal dentin with fluorescence confocal laser microscopy. The push-out bond strength was then determined, and intraradicular sealing observed by using scanning electron microscopy. RESULTS: Fiber-reinforced composite resin dowels showed a closer intraradicular fit and seal in the root canal dentin, and the morphology of the apical portion of the fiber-reinforced composite resin dowels showed a sealing area with the gutta percha in the apical portion. The mean thickness of the resin cement layer was significantly reduced for the fiber-reinforced composite resin dowels compared with the prefabricated dowels (P<.05) in the cervical area (197.0 µm versus 311.0 µm) and in the apical portion of the root canal (57.3 µm versus 131.6 µm). The mean push-out strength was higher for the fiber-reinforced composite resin dowels (22.98 N/mm2) than that for the prefabricated dowels (16.49 N/mm2) (P<.05). CONCLUSIONS: The morphological fit of fiber-reinforced composite resin dowels provides better intraradicular sealing in the cervical and apical portions, reducing the resin cement thickness. The increased push-out strength can therefore be assumed to result from increased frictional retention compared with prefabricated dowels.

Stopping Hospital Infections With Environmental Services (SHINE): A Cluster-randomized Trial of Intensive Monitoring Methods for Terminal Room Cleaning on Rates of Multidrug-resistant Organisms in the Intensive Care Unit.

BACKGROUND: Multidrug-resistant organisms (MDROs) frequently contaminate hospital environments. We performed a multicenter, cluster-randomized, crossover trial of 2 methods for monitoring of terminal cleaning effectiveness. METHODS: Six intensive care units (ICUs) at 3 medical centers received both interventions sequentially, in randomized order. Ten surfaces were surveyed each in 5 rooms weekly, after terminal cleaning, with adenosine triphosphate (ATP) monitoring or an ultraviolet fluorescent marker (UV/F). Results were delivered to environmental services staff in real time with failing surfaces recleaned. We measured monthly rates of MDRO infection or colonization, including methicillin-resistant Staphylococcus aureus, Clostridioides difficile, vancomycin-resistant Enterococcus, and MDR gram-negative bacilli (MDR-GNB) during a 12-month baseline period and sequential 6-month intervention periods, separated by a 2-month washout. Primary analysis compared only the randomized intervention periods, whereas secondary analysis included the baseline. RESULTS: The ATP method was associated with a reduction in incidence rate of MDRO infection or colonization compared with the UV/F period (incidence rate ratio [IRR] 0.876; 95% confidence interval [CI], 0.807-0.951; P = .002). Including the baseline period, the ATP method was associated with reduced infection with MDROs (IRR 0.924; 95% CI, 0.855-0.998; P = .04), and MDR-GNB infection or colonization (IRR 0.856; 95% CI, 0.825-0.887; P < .001). The UV/F intervention was not associated with a statistically significant impact on these outcomes. Room turnaround time increased by a median of 1 minute with the ATP intervention and 4.5 minutes with UV/F compared with baseline. CONCLUSIONS: Intensive monitoring of ICU terminal room cleaning with an ATP modality is associated with a reduction of MDRO infection and colonization.

Occurrence and transmission potential of asymptomatic and presymptomatic SARS-CoV-2 infections: Update of a living systematic review and meta-analysis.

BACKGROUND: Debate about the level of asymptomatic Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection continues. The amount of evidence is increasing and study designs have changed over time. We updated a living systematic review to address 3 questions: (1) Among people who become infected with SARS-CoV-2, what proportion does not experience symptoms at all during their infection? (2) What is the infectiousness of asymptomatic and presymptomatic, compared with symptomatic, SARS-CoV-2 infection? (3) What proportion of SARS-CoV-2 transmission in a population is accounted for by people who are asymptomatic or presymptomatic? METHODS AND FINDINGS: The protocol was first published on 1 April 2020 and last updated on 18 June 2021. We searched PubMed, Embase, bioRxiv, and medRxiv, aggregated in a database of SARS-CoV-2 literature, most recently on 6 July 2021. Studies of people with PCR-diagnosed SARS-CoV-2, which documented symptom status at the beginning and end of follow-up, or mathematical modelling studies were included. Studies restricted to people already diagnosed, of single individuals or families, or without sufficient follow-up were excluded. One reviewer extracted data and a second verified the extraction, with disagreement resolved by discussion or a third reviewer. Risk of bias in empirical studies was assessed with a bespoke checklist and modelling studies with a published checklist. All data syntheses were done using random effects models. Review question (1): We included 130 studies. Heterogeneity was high so we did not estimate a mean proportion of asymptomatic infections overall (interquartile range (IQR) 14% to 50%, prediction interval 2% to 90%), or in 84 studies based on screening of defined populations (IQR 20% to 65%, prediction interval 4% to 94%). In 46 studies based on contact or outbreak investigations, the summary proportion asymptomatic was 19% (95% confidence interval (CI) 15% to 25%, prediction interval 2% to 70%). (2) The secondary attack rate in contacts of people with asymptomatic infection compared with symptomatic infection was 0.32 (95% CI 0.16 to 0.64, prediction interval 0.11 to 0.95, 8 studies). (3) In 13 modelling studies fit to data, the proportion of all SARS-CoV-2 transmission from presymptomatic individuals was higher than from asymptomatic individuals. Limitations of the evidence include high heterogeneity and high risks of selection and information bias in studies that were not designed to measure persistently asymptomatic infection, and limited information about variants of concern or in people who have been vaccinated. CONCLUSIONS: Based on studies published up to July 2021, most SARS-CoV-2 infections were not persistently asymptomatic, and asymptomatic infections were less infectious than symptomatic infections. Summary estimates from meta-analysis may be misleading when variability between studies is extreme and prediction intervals should be presented. Future studies should determine the asymptomatic proportion of SARS-CoV-2 infections caused by variants of concern and in people with immunity following vaccination or previous infection. Without prospective longitudinal studies with methods that minimise selection and measurement biases, further updates with the study types included in this living systematic review are unlikely to be able to provide a reliable summary estimate of the proportion of asymptomatic infections caused by SARS-CoV-2. REVIEW PROTOCOL: Open Science Framework (https://osf.io/9ewys/).