Relevance of Tacrolimus Trough Concentration and Hepatitis E virus Genetic Changes in Kidney Transplant Recipients With Chronic Hepatitis E.

Introduction: Hepatitis E virus (HEV) can cause chronic infection (≥3 months) and cirrhosis in immunocompromised patients, especially kidney transplant recipients. Low alanine aminotransferase (ALT) levels and high HEV intrahost diversity have previously been associated with evolution toward chronicity in these patients. We hypothesized that additional clinical and viral factors could be associated with the risk of chronic HEV infection. Methods: We investigated a series of 27 kidney transplant recipients with HEV infection, including 20 patients with chronic hepatitis E. Results: High tacrolimus trough concentration at diagnosis was the most relevant marker associated with chronic hepatitis E (9.2 vs. 6.4 ng/ml, P = 0.04). Most HEV genetic changes selected during HEV infection were compartmentalized between plasma and feces. Conclusion: This compartmentalization highlights the diversity and complexity of HEV replication compartments. Tacrolimus trough concentration at diagnosis of HEV infection could allow an early identification of patients at high risk of chronic hepatitis E and guide treatment initiation.

A pig model of chronic hepatitis E displaying persistent viremia and a downregulation of innate immune responses in the liver.

BACKGROUND: Hepatitis E virus (HEV) is a zoonotic virus transmitted by pig meat and responsible for chronic hepatitis E in immunocompromised patients. It has proved challenging to reproduce this disease in its natural reservoir. We therefore aimed to develop a pig model of chronic hepatitis E to improve the characterization of this disease. METHODS: Ten pigs were treated with a tacrolimus-based regimen and intravenously inoculated with HEV. Tacrolimus trough concentration, HEV viremia, viral diversity, innate immune responses, liver histology, clinical disease and biochemical markers were monitored for 11 weeks post-infection (p.i.). RESULTS: HEV viremia persisted for 11 weeks p.i. HEV RNA was detected in the liver, small intestine, and colon at necropsy. Histological analysis revealed liver inflammation and fibrosis. Several mutations selected in the HEV genome were associated with compartmentalization in the feces and intestinal tissues, consistent with the hypothesis of extrahepatic replication in the digestive tract. Antiviral responses were characterized by a downregulation of IFN pathways in the liver, despite an upregulation of RIG-I and ISGs in the blood and liver. CONCLUSIONS: We developed a pig model of chronic hepatitis E that reproduced the major hallmarks of this disease. This model revealed a compartmentalization of HEV genomes in the digestive tract and a downregulation of innate immune responses in the liver. These original features highlight the relevance of our model for studies of the pathogenesis of chronic hepatitis E and for validating future treatments.

The Performance Diagnostic Checklist-Human Services (1.1): An initial assessment of validity and reliability.

The Performance Diagnostic Checklist-Human Services (PDC-HS) is an assessment used to identify variables contributing to staff performance concerns in human-service settings. In the current study, we introduce and assess the test validity, interrater reliability, and test-retest reliability of the PDC-HS (1.1), a revised version of the assessment that included revised instructions, questions, and intervention planning references. We measured the psychometric properties of the revised assessment by analyzing answers obtained from watching video vignettes of simulated interviews between consultants and a supervisor. Twenty-one participants watched the vignettes and completed the PDC-HS (1.1) based on the answers provided during the interview. We also included an item analysis to identify questions on which participants made errors and an intervention selection task to assess whether participants selected an appropriate intervention to target the indicated domain. The results support the use of the PDC-HS (1.1) in human services settings.

A detailed examination of reporting procedural fidelity in the Journal of Applied Behavior Analysis.

Few reviews on procedural fidelity-the degree to which procedures are implemented as designed-provide details to gauge the quality of fidelity reporting in behavior-analytic research. This review focused on experiments in the Journal of Applied Behavior Analysis (2006-2021) with "integrity" or "fidelity" in the abstract or body. When fidelity data were collected, the coders characterized measurement details (e.g., description of calculation, report of single or multiple values, frequency of fidelity checks, checklist use). The researchers found increasing trends in describing the calculation(s), reporting multiple values, and stating the frequency of measurement. Few studies described using a checklist. Most studies reported fidelity as a percentage, with high obtained values (M = 97%). When not collecting fidelity data was stated as a limitation, authors were unlikely to provide a rationale for the omission. We discuss recommendations for reporting procedural fidelity to increase the quality of and transparency in behavior-analytic research.

Procedural parameters in equivalence-based instruction with individuals diagnosed with autism: A call for systematic research.

Equivalence-based instruction (EBI) is an efficient and efficacious methodology to establish equivalence classes that has been used to teach various academic skills to neurotypical adults. Although previous reviews confirmed the utility of EBI with participants with developmental disabilities, it is unclear whether certain procedural parameters are associated with positive equivalence outcomes. We extended previous reviews by categorizing studies that used EBI with individuals diagnosed with autism spectrum disorder and assessed whether any procedural parameters were associated with better equivalence responding. Due to the wide variability of procedural parameters in EBI research, the best procedural permutations to form equivalence classes with individuals diagnosed with autism spectrum disorder are still unknown. Thus, this paper serves as a call to action for applied researchers. Specifically, we encourage and invite researchers to systematically investigate the necessary variables or combination of variables that may lead to successful equivalence class formation.

A parametric analysis of procedural fidelity errors following mastery of a task: A translational study.

Procedural fidelity is defined as the extent to which the independent variable is implemented as prescribed. Research using computerized tasks has shown that fidelity errors involving consequences for behavior can hinder skill acquisition. However, studies examining the effects of these errors once skills have been mastered are lacking. Thus, this translational study investigated the effects of varying levels of fidelity following mastery of a computerized arbitrary matching-to-sample task. A group design (consisting of five groups) was used in which college students initially completed 250 trials during which no programmed errors (i.e., perfect fidelity) were arranged, followed by an additional 250 trials with consequences delivered across various levels of fidelity (i.e., 20, 40, 60, 80, and 100% of trials administered without errors). The results showed that participants assigned to higher fidelity conditions performed better (on average). These results extended the findings of previous studies by demonstrating how errors involving consequences affect behavior across various stages of learning.

Trends in Reporting Procedural Integrity: A Comparison.

Procedural integrity refers to the extent to which an independent variable is implemented as described. Measuring procedural integrity is one important factor when considering internal and external validity of experiments. Experimental articles in behavior-analytic journals have rarely reported procedural-integrity data. The purpose of this study was to update previous reviews of whether articles published in the Journal of Applied Behavior Analysis reported procedural integrity, spanning a period from 1980 to 2020, and compare reporting in JABA to recent reviews of studies published in Behavior Analysis in Practice (2008-2019) and the Journal of Organizational Behavior Management (2000-2020). Procedural integrity continues to be underreported across all three journals, but an increasing trend in reporting procedural integrity is evident in the Journal of Applied Behavior Analysis and Behavior Analysis in Practice. In addition to our recommendations and implications for research and practice, we provide examples and resources to assist researchers and practitioners with recording and reporting integrity data.

On a persisting curious double standard in behavior analysis: Behavioral scholars' perspectives on procedural fidelity.

Procedural fidelity is the extent to which independent variables are implemented as designed. Despite 40 years of discussion about the importance of procedural fidelity for behavioral research, reporting of fidelity data remains an uncommon practice in behavior-analytic journals. Researchers have speculated about reasons for underreporting, but the perspectives of scholars about when reporting is warranted or necessary have not yet been explored. Thus, the purpose of this study was to evaluate possible reasons for infrequent reporting of fidelity data in behavior-analytic studies. To address this purpose, we conducted focus groups with scholars in applied behavior analysis. Five themes emerged regarding why procedural fidelity data are not typically reported. We provide a discussion about how these themes are interrelated and offer suggestions and recommendations to assist with the collection and reporting of fidelity data.

The Performance Diagnostic Checklist - Human Services: Guidance for Assessment Administration.

The Performance Diagnostic Checklist - Human Services (PDC-HS) is an assessment designed to assess the environmental variables contributing to employee performance concerns in human-service settings. Recent research has demonstrated that interventions indicated by the PDC-HS result in improved employee performance across several human-service settings and that the assessment has acceptable reliability and validity. Although PDC-HS-indicated interventions have been effective at increasing employee performance, there is a need for additional guidance when using the assessment given the limited nature of the original administration guidelines. Thus, the purpose of the current manuscript is to introduce additional guidance for use of the PDC-HS across a variety of situations.

Treatment Integrity Reporting in Behavior Analysis in Practice 2008-2019.

Treatment integrity is the extent to which procedures are implemented in a manner consistent with their prescribed protocols and is necessary for reaching accurate conclusions regarding functional relations between dependent (i.e., behavior) and independent (i.e., the environment) variables. Several studies assessing the frequency that studies report treatment integrity have been conducted. However, no review has included articles from Behavior Analysis in Practice. Thus, the current study reviewed Behavior Analysis in Practice between 2008 and 2019 to assess the frequency of studies reporting treatment integrity data. A total of 193 articles consisting of 205 studies met the inclusionary criteria for this review. Ninety-six studies (46.83%) reported treatment integrity data, compared to 193 (94.15%) that provided interobserver agreement data. Additionally, 98 studies (47.80%) were considered high risk for treatment implementation inaccuracies. Recommendations and implications for research and practice are discussed.

Assessing the efficacy of and preference for positive and corrective feedback.

Feedback is an effective strategy for improving performance and consists of multiple characteristics. One characteristic that can influence feedback efficacy is its nature (whether feedback is positive or corrective) and little is known about the conditions under which individuals may prefer corrective over positive feedback. Thus, the purpose of this study was to assess the efficacy of and preference for positive and corrective feedback during the acquisition of novel tasks. In the first phase, participants received either positive, corrective, or no feedback across three novel tasks. Participants only mastered the task in which they received corrective feedback. In the second phase, participants chose to receive either positive or corrective feedback after completing trials of the previous phase's control task. All participants chose to receive corrective feedback more frequently than positive feedback. We discuss the implications of the results for feedback delivery in the workplace and provide suggestions for future research.

Evolution and phenotypic characterization of whole HBV genome in compliant patients experiencing unexplained entecavir treatment failure.

Entecavir treatment failure can be observed in compliant patients despite an absence of detectable resistance mutations by Pol/RT Sanger sequencing. We hypothesized that these unexplained treatment failures could rely on other mechanisms of viral resistance, especially on mutations selected outside of the Pol/RT domain. Partial virological response to entecavir was observed in three patients treated with immunosuppressive drugs, without selection of Pol/RT resistance mutations. Mutations selected in the whole HBV genome during entecavir treatment and potentially associated with resistance were searched for using deep sequencing and characterized using a phenotypic resistance assay. Mutations Q206K (pre-core/core), Q120K (pre-S1/pre-S2, T-cell epitope) and A300E (spacer domain) were selected during entecavir treatment in patient #1 but were not associated with an increased level of resistance to entecavir or an increase in HBV replication capacity. Core promoter mutations T1753G, A1762T and G1764A were present as major mutations before and after treatment in patient #1. HBs Ag immune escape mutations were present as major mutations before and after treatment in patients #2 (sK122R, sT126I, sP127S and sG145R) and #3 (sM133I). We demonstrated that PVR to entecavir does not require selection of any resistance mutation in the whole HBV genome. Our results demonstrate that major mutations can be selected outside of the Pol/RT domain before or during entecavir treatment. These mutations could contribute to entecavir treatment failure by other mechanisms than an increased level of resistance.

Synthesis and Antiviral Evaluation of (1,4-Disubstituted-1,2,3-Triazol)-(E)-2-Methyl-but-2-Enyl Nucleoside Phosphonate Prodrugs.

A series of hitherto unknown (1,4-disubstituted-1,2,3-triazol)-(E)-2-methyl-but-2-enyl nucleosides phosphonate prodrugs bearing 4-substituted-1,2,3-triazoles were prepared in a straight approach through an olefin acyclic cross metathesis as the key synthetic step. All novel compounds were evaluated for their antiviral activities against HBV, HIV and SARS-CoV-2. Among these molecules, only compound 15j, a hexadecyloxypropyl (HDP)/(isopropyloxycarbonyl-oxymethyl)-ester (POC) prodrug, showed activity against HBV in Huh7 cell cultures with 62% inhibition at 10 µM, without significant cytotoxicity (IC50 = 66.4 µM in HepG2 cells, IC50 = 43.1 µM in HepG2 cells) at 10 µM.

The Role of Phosphatidylinositol Phosphate Kinases during Viral Infection.

Phosphoinositides account for only a small proportion of cellular phospholipids, but have long been known to play an important role in diverse cellular processes, such as cell signaling, the establishment of organelle identity, and the regulation of cytoskeleton and membrane dynamics. As expected, given their pleiotropic regulatory functions, they have key functions in viral replication. The spatial restriction and steady-state levels of each phosphoinositide depend primarily on the concerted action of specific phosphoinositide kinases and phosphatases. This review focuses on a number of remarkable examples of viral strategies involving phosphoinositide kinases to ensure effective viral replication.

Differential utilization of CD4+ by transmitted/founder and chronic envelope glycoproteins in a MSM HIV-1 subtype B transmission cluster.

OBJECTIVE: HIV-1 transmission leads to a genetic bottleneck, with one or a few variants of the donor quasispecies establishing an infection in the new host. We aimed to characterize this bottleneck in more detail, by comparing the properties of HIV envelope glycoproteins from acute and chronic infections within the particular context of a male-to-male transmission cluster. DESIGN: We compared the genotypic and phenotypic properties of envelope glycoproteins from viral variants derived from five study participants from the same transmission cluster. METHODS: We used single-genome amplification to generate a collection of full-length env sequences. We then constructed pseudotyped viruses expressing selected Env variants from the quasispecies infecting each study participant and compared their infectivities and sensitivities to various entry inhibitors. RESULTS: The genotypic analyses confirmed the genetic bottleneck expected after HIV transmission, with a limited number of variants identified in four study participants during acute infection. However, the transmitted sequences harbored no evident common signature and belonged to various genetic lineages. The phenotypic analyses revealed no difference in infectivity, susceptibility to the CCR5 antagonist maraviroc, the fusion inhibitor enfurvitide or type-I interferon between viruses from participants with acute and chronic infections. The key property distinguishing transmitted viruses was a higher resistance to soluble CD4, correlated with greater sensitivity to occupation of the CD4 receptor by the anti-CD4 antibodies LM52 and SK3. CONCLUSION: These results suggest that envelope glycoproteins from transmitted/founder viruses bind CD4 less efficiently than those of viruses from chronic infections.

Revisiting HBV resistance to entecavir with a phenotypic approach.

Treatment adaptation after hepatitis B virus (HBV) treatment failure relies on genotypic resistance testing. However, the results of such tests are not always consistent with treatment response. These discrepancies may be due to differences in resistance levels between isolates with the same genotypic resistance testing profiles. We explored this hypothesis by investigating six cases of entecavir treatment failure with an integrative strategy combining genotypic and phenotypic resistance testing, medical record review and therapeutic drug monitoring. Among isolates with genotypic reduced susceptibility to entecavir, one displayed a higher level of resistance to entecavir (mean fold change in entecavir IC50 of 1 508 ± 531 vs. 318 ± 53, p = 0.008). This isolate harbored a substitution (rt250L) at a position reported to be associated with resistance (rt250V). Reversion to wild-type amino acid at this position partially restored susceptibility to entecavir, confirming that the rt250L mutation was responsible for the high level of resistance to entecavir. This is the first description of entecavir treatment failure associated with selection of the rt250L mutation without other entecavir resistance mutations. One isolate with genotypic resistance to entecavir, harboring the rt173L mutation, displayed a lower level of resistance than the other, harboring the rt202G mutation (mean fold change of 323 ± 124 vs. 6 036 ± 2 100, p = 0.20). These results suggest that isolates harboring the rt250L mutations should be considered resistant to entecavir, whereas isolates harboring the rt173L mutations should be considered to display reduced susceptibility to entecavir. An integrative approach to antiviral drug resistance in HBV would provide a more accurate assessment of entecavir treatment failures and help to improve the accuracy of genotypic testing algorithms.

Type I Phosphatidylinositol-4-Phosphate 5-Kinases α and γ Play a Key Role in Targeting HIV-1 Pr55Gag to the Plasma Membrane.

HIV-1 assembly occurs principally at the plasma membrane (PM) of infected cells. Gag polyprotein precursors (Pr55Gag) are targeted to the PM, and their binding is mediated by the interaction of myristoylated matrix domain and a PM-specific phosphoinositide, the phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P2]. The major synthesis pathway of PI(4,5)P2 involves the activity of phosphatidylinositol-4-phosphate 5-kinase family type 1 composed of three isoforms (PIP5K1α, PIP5K1ß, and PIP5K1γ). To examine whether the activity of a specific PIP5K1 isoform determines proper Pr55Gag localization at the PM, we compared the cellular behavior of Pr55Gag in the context of PIP5K1 inhibition using siRNAs that individually targeted each of the three isoforms in TZM-bl HeLa cells. We found that downregulation of PIP5K1α and PIP5K1γ strongly impaired the targeting of Pr55Gag to the PM with a rerouting of the polyprotein within intracellular compartments. The efficiency of Pr55Gag release was thus impaired through the silencing of these two isoforms, while PIP5K1ß is dispensable for Pr55Gag targeting to the PM. The PM mistargeting due to the silencing of PIP5K1α leads to Pr55Gag hydrolysis through lysosome and proteasome pathways, while the silencing of PIP5K1γ leads to Pr55Gag accumulation in late endosomes. Our findings demonstrated that, within the PIP5K1 family, only the PI(4,5)P2 pools produced by PIP5K1α and PIP5K1γ are involved in the Pr55Gag PM targeting process.IMPORTANCE PM specificity of Pr55Gag membrane binding is mediated through the interaction of PI(4,5)P2 with the matrix (MA) basic residues. It was shown that overexpression of a PI(4,5)P2-depleting enzyme strongly impaired PM localization of Pr55Gag However, cellular factors that control PI(4,5)P2 production required for Pr55Gag-PM targeting have not yet been characterized. In this study, by individually inhibiting PIP5K1 isoforms, we elucidated a correlation between PI(4,5)P2 metabolism pathways mediated by PIP5K1 isoforms and the targeting of Pr55Gag to the PM of TZM-bl HeLa cells. Confocal microscopy analyses of cells depleted from PIP5K1α and PIP5K1γ show a rerouting of Pr55Gag to various intracellular compartments. Notably, Pr55Gag is degraded by the proteasome and/or by the lysosomes in PIP5K1α-depleted cells, while Pr55Gag is targeted to endosomal vesicles in PIP5K1γ-depleted cells. Thus, our results highlight, for the first time, the roles of PIP5K1α and PIP5K1γ as determinants of Pr55Gag targeting to the PM.

A Survey Assessing Privacy Concerns of Smart-Home Services Provided to Individuals with Disabilities.

Privacy has been identified as a primary concern among stakeholders (i.e., service recipients, advocates, administrators, family) when using technology to provide residential services to individuals in need. This paper summarizes a study that distributed a survey to agencies that provide services (e.g., clinical, recreational) and resources (e.g., advocacy groups) to people with various types of disabilities (e.g., physical, sensory, intellectual, developmental) across the United States. The results led to several recommendations about how smart-home service providers can use technology in a way that promotes client privacy. In addition, we make several suggestions for how remote staff (i.e., individuals monitoring the information gathered by technology) can assist in the process of ensuring client privacy.

Enhancing the training integrity of human service staff using pyramidal behavioral skills training.

This experiment used a pyramidal training model to evaluate the effects of behavioral skills training (BST), delivered in a 1-time group-training format, on the extent to which 25 human service staff implemented BST when training others how to implement behavioral procedures. Results indicated that (a) the training workshop increased BST integrity to mastery levels for the majority of participants with varying levels of education, organizational positions, and training experience, (b) the training effects generalized to teaching an untrained skill, and (c) high levels of BST integrity maintained at follow-up 4 to 6 weeks after training for all 3 participants with whom probes were conducted. Moreover, participants indicated high levels of satisfaction with both the training workshop and BST as a training procedure.

HIV surveillance combining an assay for identification of very recent infection and phylogenetic analyses on dried spots.

BACKGROUND: Transmitted/founder viruses isolated at the early stage of infection are indicators of the variants that are spreading within a population. The French reporting system for new HIV diagnoses is linked to a virological surveillance using dried serum spots. METHODS: We combined an immunoassay for very recent infection (less than 31 days) to a phylogenetic analysis of transmitted/founder viruses and sociodemographic information to analyze the dynamics of the HIV-1 epidemic during a 3-year period. Bayesian coalescent-based methods were used to explore the temporal and spatial dynamics of the identified clusters. RESULTS: Of 17 010 dried serum spots collected, 549 very recent infections were identified for which both env sequences and sociodemographic data were available. Non-B transmitted/founder viruses were found in 196 cases (35.7%), belonging to six subtypes and seven circulating recombinant forms. Forty-three dyads/clusters were identified (range 2-11 cases), including 107 individuals (19.5%), mainly MSM. The largest cluster involved MSM infected by a CRF02_AG variant. Reconstruction of viral migrations across time suggests that Paris was the major hub of dissemination. CONCLUSION: The study shows the feasibility of the surveillance of the HIV epidemic using this methodology. The observation of actively growing spatiotemporal clusters allows identification of specific networks that may be targets for intervention.