Longer Persistence Research Articles

Escape from the garden: spreading, effects and traits of a new risky invasive ornamental plant (Gaillardia aristata Pursh)

Ornamental plants constitute a major source of invasive species.Gaillardia aristata(great blanketflower) is planted worldwide and its escape has been reported in several European countries without ecological impact assessment on the invasive potential. As there is a markedly spreading population with invasive behaviour in Hungary, we aimed to reveal the distribution, impacts and traits ofG. aristata. We gathered occurrence data outside the gardens in Hungary, based on literature, unpublished observations by experts and our own records. We investigated the impacts of an extended population, where the species invaded sandy old-fields within a 25 km2area. Here, we compared the species richness, diversity, community composition and height of invaded and uninvaded vegetation. Furthermore, we evaluated the traits potentially associated with the invasiveness ofG. aristatain comparison with other herbaceous invasive species in the region. We found thatG. aristataoccurred mostly by casual escapes, but naturalised and invasive populations were also detected in considerable numbers.G. aristatausually appeared close to gardens and ruderal habitats, but also in semi-natural and natural grasslands and tended to spread better in sandy soils. We found lower plant species richness and Shannon diversity in the invaded sites and the invasion ofG. aristatasignificantly influenced the composition of the plant community. The trait analyses revealed that the invasive potential ofG. aristatais backed by a wide germination niche breadth, extremely long flowering period, small shoot-root ratio (large absorption and gripping surface), large seeds (longer persistence) and dispersal by epizoochory of grazing livestock (mostly by sheep), probably helping the species’ survival and spreading in the disturbed, species-poor, sandy, open habitats. These functional traits, as well as the ornamental utilisation, may act together with the aridisation of the climate and the changing land-use practices (e.g. abandoned, disturbed sites) in the success ofG. aristata. We raise awareness of the rapid transition ofG. aristatafrom ornamental plant to casual alien and then to invasive species in certain environmental conditions (i.e. sandy soils, species-poor communities, human disturbances), although it seems to be not a strong ecosystem transformer so far. Nonetheless, banning it from seed mixtures, developing eradication strategy and long-term monitoring of this species would be important to halt its spreading in time.

CAR T-Cell Persistence Correlates with Improved Outcome in Patients with B-Cell Lymphoma.

Chimeric antigen receptor (CAR) T-cell therapy has led to profound and durable tumor responses in a relevant subset of patients with relapsed/refractory (r/r) B-cell lymphomas. Still, some patients show insufficient benefit or relapse after CAR T-cell therapy. We performed a retrospective study to investigate the correlation between CAR T-cell persistence in the peripheral blood (PB) at 6 months, assessed by droplet digital PCR (ddPCR), with CAR T-cell treatment outcome. 92 patients with r/r B-cell lymphomas were treated with CD19-targeting CAR T-cell therapies at our institution between 01/2019-08/2022. Six months post-treatment, 15 (16%) patients had no detectable circulating CAR-T constructs by ddPCR. Patients with CAR T-cell persistence had a significantly higher CAR T-cell peak (5432 vs. 620 copies/ug cfDNA, p = 0.0096), as well as higher incidence of immune effector cell-associated neurotoxicity syndrome (37% vs. 7%, p = 0.0182). After a median follow-up of 8.5 months, 31 (34%) patients relapsed. Lymphoma relapses were less frequent among patients with CAR T-cell persistence (29% vs. 60%, p = 0.0336), and CAR T-cell persistence in the PB at 6 months was associated with longer progression-free survival (PFS) (HR 2.79, 95% CI: 1.09-7.11, p = 0.0319). Moreover, we observed a trend towards improved overall survival (OS) (HR 1.99, 95% CI: 0.68-5.82, p = 0.2092) for these patients. In our cohort of 92 B-cell lymphomas, CAR T-cell persistence at 6 months was associated with lower relapse rates and longer PFS. Moreover, our data confirm that 4-1BB-CAR T-cells have a longer persistence as compared to CD-28-based CAR T-cells.

Nanotechnological approaches for management of soil-borne plant pathogens.

Soil borne pathogens are significant contributor of plant yield loss globally. The constraints in early diagnosis, wide host range, longer persistence in soil makes their management cumbersome and difficult. Therefore, it is crucial to devise innovative and effective management strategy to combat the losses caused by soil borne diseases. The use of chemical pesticides is the mainstay of current plant disease management practices that potentially cause ecological imbalance. Nanotechnology presents a suitable alternative to overcome the challenges associated with diagnosis and management of soil-borne plant pathogens. This review explores the use of nanotechnology for the management of soil-borne diseases using a variety of strategies, such as nanoparticles acting as a protectant, as carriers of actives like pesticides, fertilizers, antimicrobials, and microbes or by promoting plant growth and development. Nanotechnology can also be used for precise and accurate detection of soil-borne pathogens for devising efficient management strategy. The unique physico-chemical properties of nanoparticles allow greater penetration and interaction with biological membrane thereby increasing its efficacy and releasability. However, the nanoscience specifically agricultural nanotechnology is still in its toddler stage and to realize its full potential, extensive field trials, utilization of pest crop host system and toxicological studies are essential to tackle the fundamental queries associated with development of commercial nano-formulations.

Metabolic Disposition and Elimination of Tritum-Labeled Sulfamethoxazole in Pigs, Chickens and Rats.

Sulfamethoxazole (SMZ), as a sulfa antibiotic, is often used in the treatment of various infectious diseases in animal husbandry. At present, SMZ still has many unresolved problems in the material balance, metabolic pathways, and residual target tissues in food animals. Therefore, in order to solve these problems, the metabolism, distribution, and elimination of SMZ is investigated in pigs, chickens, and rats by radioactive tracing methods, and the residue marker and target tissue of SMZ in food animals were determined, providing a reliable basis for food safety. After a single administration of [3H]-SMZ (rats and pigs by intramuscular injection and chickens by oral gavage), the total radioactivity was rapidly excreted, with more than 93% of the dose excreted within 14 days in the three species. Pigs and rats had more than 75% of the administered volume recovered by urine. After 7 days of continuous administration, within the first 6 h, radioactivity was found in almost all tissues. The highest radioactivity and longest persistence in pigs was in the liver, while in chickens it was in the liver and kidneys, most of which was removed within 14 days. A total of six, three and three metabolites were found in chickens, rats and pigs, respectively. N4-acetyl-sulfamethoxazole (S1) was the main metabolite of SMZ in rats, pigs and chickens. The radioactive substance with the longest elimination half-life is sulfamethoxazole (S0), so S0 was suggested to be the marker residue in pigs and chickens.

Persistence, leaching and associated toxicity risks of insecticide pyriproxyfen in soil ecosystem.

Pyriproxyfen is a pyridine-based insecticide used for pest control in fruits and vegetables. It is a potent endocrine disruptor and hormone imitator. Considering its potential hazards to non-target organisms and the associated environment, a lab study was conducted for assessing persistence, mobility in sandy loam soil and associated risk to various non-target organisms and soil enzymes. Pyriproxyfen formulation was applied at 0.05 and 0.10 µg g-1 soil which was equivalent to recommended and double dose of 100 and 200 g a.i. ha-1, respectively. Three methods namely QuEChERS, liquid-solid extraction (LSE) and matrix solid phase dispersion (MSPD) were compared for achieving efficient sample preparation. MSPD was applied for final analysis as it gave better recoveries (94.2 to 104.3%) over other methods with limits of detection and quantification (LOD and LOQ) as 0.0001 and 0.0005 µg g-1, respectively. Dissipation followed first order kinetics with half-lives of 7.6 and 8.2 days in both doses but residues retained over 45 days in soil. Leaching studies conducted at 50 and 100 µg of pyriproxyfen showed extremely poor leaching potential. Retention of over 90% residues in top 5 cm soil surface indicated minimal threat of ground and surface water contamination. Toxicological study demonstrated very different behaviour toward different enzymatic activities. Pyriproxyfen was relatively toxic for alkaline phosphatase and fluorescein diacetate hydrolase enzymes. β-glucosidase activity was triggered whereas arylsulfatase activity remained unaffected. Unacceptable risk to soil invertebrates at double dose application clearly indicated that its longer persistence in soil could be toxic to other non-target organisms and needs further investigations.

The Different Relationships between the ENSO Spring Persistence Barrier and Predictability Barrier

Abstract In this paper, we investigate the relationship between the El Niño–Southern Oscillation (ENSO) spring persistence barrier (PB) and predictability barrier (PD) and apply it to explain the interdecadal modulation of ENSO prediction skill using the anomaly correlation coefficient (ACC). Previous studies showed that a longer persistence (i.e., autocorrelation) tends to produce a higher prediction skill. Using the recharge oscillator model of ENSO, both analytical and numerical solutions suggest that the predictability (i.e., ACC) is related to the persistence of sea surface temperature (SST) and cross correlation between SST and subsurface ocean heat content in the tropical Pacific. In particular, a larger damping rate in SST anomalies will lead to a lower persistence and ACC and a stronger PD. However, a shortened ENSO period, which controls the cross correlation, will lead to a lower persistence but a higher ACC associated with a weaker PD. Finally, we apply our solutions to observations and suggest that a higher ACC associated with a weaker PD after 1960 is caused by the shortened ENSO period.

Treatment persistence and adherence in people with type 2 diabetes switching to iGlarLixi vs free-dose combinations of basal insulin and glucagon-like peptide 1 receptor agonist.

BACKGROUND: Fixed-ratio combinations of basal insulin (BI) and glucagon-like peptide-1 receptor agonists (GLP-1 RAs) have greater simplicity of administration with expected improved adherence/persistence with therapy, but real-world data are lacking. OBJECTIVE: To compare medication persistence, adherence, and health care resource utilization (HRU) and costs for insulin glargine 100 U/mL and the GLP-1 RA lixisenatide (iGlarLixi) with newly initiated free-dose combinations of BI and GLP-1 RAs initiated simultaneously or sequentially. METHODS: This analysis used the US Optum Clinformatics (January 2017 to November 2019) database and included data from adults (aged ≥ 18 years) with type 2 diabetes and a glycated hemoglobin A1c (A1c) of 8% or more. Participants received iGlarLixi or free-dose combinations of BI and GLP-1 RAs prescribed simultaneously or subsequently. Participants were followed for 12 months. Cohorts were propensity score matched on baseline characteristics. The primary outcome was persistence (days on treatment without discontinuation). Secondary outcomes were adherence (proportion of days covered), change in A1c, and all-cause and diabetes-related health care resource utilization and costs. Subgroup analyses were performed for individuals with A1c levels of 9% or more. RESULTS: After propensity score matching, there were 1,357 patients in each group; groups were well balanced. In the free-dose combination group, 65.6% started on BI, then added GLP-1 RAs; 28.5% started on GLP-1 RAs, then added BI; and 5.9% started on GLP-1 RAs and BI on the same day. In the subgroup with a baseline of A1c levels of 9% or more, 952 (iGlarLixi) and 932 (free-dose combination) participants were included. A significantly higher proportion of participants in the overall population who received iGlarLixi vs free-dose combinations were persistent (44.8% vs 36.3% [hazard ratio = 1.22, 95% CI = 1.11-1.35, P < 0.001]; the median [Q1, Q3] number of persistent days was 150 [63, 360] vs 120 [60, 310]) and adherent to therapy (41.3% vs 18.7%, [odds ratio = 3.06, 95% CI = 2.57-3.65; P < 0.001]). Results for persistence in the subpopulation of participants with HbA1c levels of 9% or more were similar. Reductions in A1c from baseline were similar between iGlarLixi and the free-dose combination group (overall population: -1.2% vs -1.3%; P = 0.1913), but the number of participants in the database with follow-up A1c data was low. All-cause and diabetes-related pharmacy visits and total medication and diabetes medication pharmacy claims costs were significantly lower (all P < 0.001) for those receiving iGlarLixi vs free-dose combinations in both populations. CONCLUSIONS: In adults with type 2 diabetes, iGlarLixi was associated with longer persistence by approximately 30 days, improved adherence, and reductions in outpatient and pharmacy visits and in pharmacy costs. DISCLOSURES: This study was funded by Sanofi US. Medical writing support was provided by Barrie Anthony, PhD, CMPP, of Evidence Scientific Solutions and funded by Sanofi US. Dr Edelman has been on an advisory board and speakers' bureau for AstraZeneca, MannKind, and Xeris and on an advisory board for BrightSight and is a board member for Senseonics and Team-Type1. Mr Cassarino is on the speakers' bureau for Sanofi. Dr Kayne has been a consultant and speakers' bureau member for AstraZeneca, Bayer, Dexcom, Eli Lilly & Company, Janssen, MannKind, Novo Nordisk, and Sanofi. Dr Dex and Mr Li are employees of Sanofi. Dr Pasquel has received unrestricted research support from Dexcom, Insulet, and Merck and has been a consultant for Medscape, AI Health, Boehringer Ingelheim, and Dexcom.

Real-world medication persistence among HIV-1 patients initiating integrase inhibitor-based antiretroviral therapy in Japan

IntroductionMedication persistence has rarely been studied for integrase strand transfer inhibitor (INSTI)-based regimens among patients living HIV (PLWH) in Asia. This study investigated medication persistence for newly prescribed INSTI-based regimens in Japan by comparing single-tablet regimens (STRs) versus multiple-tablet regimens (MTRs), based on the Medical Data Vision database. MethodsAdult PLWH with ≥2 claims for antiretroviral therapy (ART) of interest between 1 January 2017 and 30 June 2018 were included if they had a ≥3-month continuous enrolment prior to the index date and a ≥6-month follow-up after the index date. Medication persistence was measured as the duration from initiation to discontinuation of the prescribed INSTI-based regimen. ResultsOverall, 487 patients were included, with 220 in the STR cohort and 267 in the MTR cohort. Persistence was longer in the STR cohort than in the MTR cohort (mean days on the index regimens: 384.2 vs. 317.3, P < 0.001). MTRs were associated with a higher risk of discontinuation than STRs (hazard ratio [HR], 1.72; 95% confidence interval [CI], 1.18–2.52; P = 0.005). Other factors that were associated with discontinuation were backbone (emtricitabine/tenofovir disoproxil fumarate vs. emtricitabine/tenofovir alafenamide: HR, 5.64; 95% CI, 3.68–8.66; P < 0.001), third agent (raltegravir vs. elvitegravir/cobicistat: HR, 2.06; 95% CI, 1.10–3.86; P = 0.024), age (HR, 1.02; 95% CI, 1.01–1.03; P = 0.007), and the number of non-ART index medications (HR, 1.16; 95% CI, 1.12–1.21; P < 0.001). ConclusionsAmong PLWH newly prescribed an INSTI-based regimen in Japan, STRs were associated with longer persistence than MTRs.

Treatment initiation and utilization patterns of pharmacotherapies for early-onset idiopathic restless legs syndrome

Objective/backgroundRestless legs syndrome (RLS) is a complex condition associated with circadian rhythm that disrupts sleep and can cause multisystemic consequences. This study assesses pharmacotherapy treatment initiation, estimates annual treatment prevalence, and assesses treatment patterns for early-onset idiopathic RLS. MethodsWe used the MarketScan Commercial Claims Database from 2012 to 2019 to conduct a new user retrospective cohort study. Annual treatment prevalence was calculated from a cross-sectional sample. Newly diagnosed adults with early-onset (18–44 years) idiopathic RLS who initiated on and off-label gabapentinoids, dopamine agonists, or levodopa/carbidopa were included. Among monotherapy users who had one year of insurance enrollment, treatment patterns (single fill, continuous use of initiated therapy, switching, and add-on therapy) were examined and mean time on the initial treatment (as a measure of persistence) was calculated. ResultsIn total, 6,828 patients were initiated on monotherapy treatment for early-onset idiopathic RLS in which 4,638 met all inclusion criteria. In 2019, annual prevalence of monotherapy treatment of diagnosed patients for ropinirole was 171.3/1,000 patients; 85.0/1,000 patients for pramipexole; and 132.1/1,000 patients for gabapentin. Overall, 22.3% (n = 1,033) of patients maintained their initiated pharmacotherapy for the entire year. Rotigotine had the longest persistence (mean 185.4 [161.4 SD] days) but this user group was the smallest (n = 29). Gabapentin enacarbil, pregabalin, and rotigotine use was low (2.8% total). ConclusionRopinirole, pramipexole, and gabapentin were initiated most often for early-onset idiopathic RLS. FDA-approved agents for RLS, including gabapentin enacarbil and rotigotine, were used less frequently. In general, persistence was low for all RLS study drugs examined.

Joint estimation of selection intensity and mutation rate under balancing selection with applications to HLA.

A composite likelihood method is introduced for jointly estimating the intensity of selection and the rate of mutation, both scaled by the effective population size, when there is balancing selection at a single multi-allelic locus in an isolated population at demographic equilibrium. The performance of the method is tested using simulated data. Average estimated mutation rates and selection intensities are close to the true values but there is considerable variation about the averages. Allowing for both population growth and population subdivision does not result in qualitative differences but the estimated mutation rates and selection intensities do not in general reflect the current effective population size. The method is applied to 3 class I (HLA-A, HLA-B, and HLA-C) and 2 class II loci (HLA-DRB1 and HLA-DQA1) in the 1000 Genomes populations. Allowing for asymmetric balancing selection has only a slight effect on the results from the symmetric model. Mutations that restore symmetry of the selection model are preferentially retained because of the tendency of natural selection to maximize average fitness. However, slight differences in selective effects result in much longer persistence time of some alleles. Trans-species polymorphism, which is characteristic of major-histocompatibility loci in vertebrates, is more likely when there are small differences in allelic fitness than when complete symmetry is assumed. Therefore, variation in allelic fitness expands the range of parameter values consistent with observations of trans-species polymorphism.

Infliximab Treatment Persistence among Japanese Patients with Chronic Inflammatory Diseases: A Retrospective Japanese Claims Data Study.

Infliximab (IFX) has contributed to the treatment of several chronic inflammatory diseases, including Crohn's disease (CD), ulcerative colitis (UC), psoriasis (Pso), and rheumatoid arthritis (RA). However, the loss of response in some patients with long-term IFX therapy has been a major problem. Randomized controlled trials (RCTs) are limited in their short duration and lack of generalizability to the real-world population. We aimed to describe the persistence rates of IFX therapy to estimate its long-term effectiveness in clinical practice. Claims data from the Japan Medical Data Center database from January 2005 to June 2017 were used. The study population was identified based on the International Classification of Diseases, 10th Revision and the Anatomical Therapeutic Chemical Classification System. The 5-year persistence rates of IFX therapy were estimated using the Kaplan-Meier method. Overall, 281, 235, 41, and 222 patients with CD, UC, Pso, and RA, respectively, were selected. The 5-year persistence rates for IFX claims were 62.9, 38.9, 22.1, and 28.1% in patients with CD, UC, Pso, and RA, respectively. Patients with CD and UC administered IFX beyond the median dose had higher persistence rates. In patients with RA, female sex and no prior use of other biologics were associated with longer persistence. In conclusion, IFX persistence rates differed across chronic inflammatory diseases, which did not correspond to the results of the major RCTs. Factors associated with longer IFX persistence were identified in each disease group. Our findings may provide useful information to facilitate the proper use of IFX.

Exploring sandy vegetation sensitivities to water storage in China’s arid and semi-arid regions

Hulunbuir, Otindag, Horqin and Mu Us are the four greater sandy lands in China. They are typical ecologically fragile areas with limited ability to resist interference and are susceptible to changes in the external environment. With climate change in recent decades, these areas have experienced severe land desertification and decreased groundwater levels. The ecological environment is at risk of further deterioration, and continuous vegetation restoration and management should be adapted to the local climate and maintain the water balance with natural precipitation. This paper combines multi-source satellite observations to explore the relationship between vegetation growth and water supply. The results show four main points. (1) Although the vegetation growth in Otindag and Horqin had degraded before 2010, it improved after 2010. The vegetation growth and soil moisture (SM) of the four sandy lands both generally increased from 2003 to 2016. The SM of the Horqin increased significantly (84% anomaly > 1.4 × 10−2 cm3/cm3). Other than the Hulunbuir, the other sandy lands show a downward trend in terrestrial water storage (TWS), especially the Mu Us declined the most (97% anomaly < −40 mm). (2) Solar-induced chlorophyll fluorescence (SIF) has specific advantages in characterizing the sandy vegetation productivity and water composition changes compared to vegetation index (EVI) or gross primary productivity (GPP). (3) The regional pattern of the vegetation-moisture relationship is mainly affected by the precipitation-driven SM and depends on the types of underlying vegetation functions. (4) Both SM and TWS have approximately one-month persistence in the four sandy lands, especially the Mu Us has the longest persistence (0.94 months and 1.1 months). The persistence in the early period is generally longer than that in the late period, implying that the resilience of water components after the mutation year is stronger than before it. This study provides a scientific basis for the management and restoration of sandy lands. It is recommended that different vegetation restoration and protection strategies should be developed for specific sandy land moisture conditions in arid/semi-arid regions of northern China, which includes the improvement of water use efficiency to maintain soil moisture and the reduction of excessive groundwater uses.

Abstract PO-255: Cervical cancer prevention and HPV self-sampling awareness and acceptability among women living with HIV: A qualitative investigation from the patients' and providers' perspectives

Abstract Background: Women living with HIV (WLH) bear a disproportionate risk of invasive cervical cancer due to greater incidence and longer persistence of high-risk HPV infection. Emerging strategies that involve women collecting their own cervicovaginal sample as an alternative to traditional office-based HPV screening may be a promising approach to reach women who face several obstacles to timely care, such as WLH. Little is known, however, about the facilitators and barriers to HPV self-sampling uptake that may be unique to this particularly vulnerable population. Purpose: This study describes their knowledge, experiences, and needs regarding cervical cancer screening, specifically HPV self-sampling, and seeks to reconcile it with the views of their providers' who have been proven to have great influences on their patients' decision-making processes. Methods: In this qualitative study, we recruited 10 providers and 39 WLH from the Washington DC metropolitan region to participate in semi-structured interviews and focus group discussions. Emergent themes were assessed using an inductive process, employing an open coding method. Results: Knowledge of cervical cancer and HPV was generally limited among WLH; and as expressed by the providers, was often fueled by proximity to someone affected by cervical cancer or personal experience. Most WLH were not familiar with HPV self-sampling: but despite some of the providers' skepticism, expressed their willingness to try it. While some providers worried that it would be an added burden, the WLH highlighted convenience, ease of use, and affordability as facilitators to the uptake of HPV self-sampling. Conclusions: The experiences identified herein may be used to (1) inform tailored interventions designed to increase cervical cancer prevention among under-screened WLH and (2) guide patient-centered communication strategies to improve engagement, patient satisfaction, quality of life, and health outcomes among vulnerable populations. Citation Format: Daisy Le, Annie Coriolan Ciceron, Min (Jaime) Jeong Jeon, Jose Bordon, Jeanne Jordan, Anne Monroe. Cervical cancer prevention and HPV self-sampling awareness and acceptability among women living with HIV: A qualitative investigation from the patients' and providers' perspectives [abstract]. In: Proceedings of the AACR Virtual Conference: 14th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2021 Oct 6-8. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr PO-255.

Porous surfaces: stability and recovery of coronaviruses.

The role of indirect contact in the transmission of SARS-CoV-2 is not clear. SARS-CoV-2 persists on dry surfaces for hours to days; published studies have largely focused on hard surfaces with less research being conducted on different porous surfaces, such as textiles. Understanding the potential risks of indirect transmission of COVID-19 is useful for settings where there is close contact with textiles, including healthcare, manufacturing and retail environments. This article aims to review current research on porous surfaces in relation to their potential as fomites of coronaviruses compared to non-porous surfaces. Current methodologies for assessing the stability and recovery of coronaviruses from surfaces are also explored. Coronaviruses are often less stable on porous surfaces than non-porous surfaces, for example, SARS-CoV-2 persists for 0.5 h-5 days on paper and 3-21 days on plastic; however, stability is dependent on the type of surface. In particular, the surface properties of textiles differ widely depending on their construction, leading to variation in the stability of coronaviruses, with longer persistence on more hydrophobic materials such as polyester (1-3 days) compared to highly absorbent cotton (2 h-4 days). These findings should be considered where there is close contact with potentially contaminated textiles.

Excretion and Residual Concentration Correlations of Salbutamol Between Edible Tissues and Living Samples in Pigs and Goats.

Illegal use of salbutamol (SAL), a β-adrenergic leanness-enhancing agent, has posed potential threat to human health in China. The excretion and depletion of SAL in pigs and goats were investigated, and the concentration correlations between edible tissues and living samples were analyzed to find out a suitable living sample for pre-slaughter monitoring of SAL in pigs and goats. After a single oral dosage of 1.2 mg/kg SAL, approximately 70% of the dose was excreted by pigs and goats from their excreta. When pigs and goats were supplied feed containing SAL (20 mg/kg) for 14 consecutive days, high concentrations of SAL were observed in the liver and kidneys, and the longest persistence was observed in hair. Unlike pigs, SAL was presented primarily as conjugated SAL in goats. Excellent concentration correlations of SAL were observed between urine and edible tissues both in pigs and goats, and in addition, good correlations also were found between hair and edible tissues in pigs and between feces and edible tissues in goats. Hence, urine and hair could accurately predict SAL concentrations in edible tissues of pigs, whereas feces and urine were satisfactory for predicting SAL concentrations in edible tissues of goats. These data make it possible for pre-slaughter monitoring of SAL residues in the edible tissues of pigs and goats.

Tri-Specific CD19xCD20xCD22 VHH CAR-T Cells (LCAR-AIO) Eradicate Antigen-Heterogeneous B Cell Tumors, Enhance Expansion, and Prolong Persistence in Preclinical In Vivo Models

Introduction: Anti-CD19 CAR-T therapy has achieved remarkable treatment efficacy in B cell lymphoma. However, targeting CD19 antigen alone can only benefit about half of patients with B cell malignancies. The FDA-approved CD19 CAR-T therapies all use same binder, which is murine FMC63 scFv targeting CD19 and up to 39%-88% of patients have relapsed. Possible mechanisms of relapse include mutations or downregulation of the targeted antigen, CD19, however, the targetable expression of CD20 and CD22 is preserved. In addition, immunogenicity against murine FMC63 scFv could have a negative impact on possible re-dosing regimen. To overcome these limitations, we designed and developed a novel tri-specific VHH CAR-T, targeting three antigens that include CD19, CD20 and CD22, for treating patients who relapsed from prior CAR-T therapies.Methods: We engineered mono-, bi-, or tri-specific VHH CAR constructs targeting CD19, CD20 and/or CD22 respectively in a lentiviral vector. The mono-, bi- or tri-specific CAR-T cells were tested against tumor lines expressing single, dual or triple antigens in an in vitro cytotoxicity assay. In addition, we evaluated the contribution of different CAR backbones, and possible combinations of scFv, VH or VHH to CAR design. We hypothesized that our lead tri-specific VHH CAR-T, LCAR-AIO, would potently inhibit tumors with heterogeneous Ag expression and prevent Ag escape. To validate this, we compared in vitro cytolytic activity and cytokine production of LCAR-AIO CAR-T to anti-CD19 FMC63 CAR-T against CD19 +CD20 +CD22 + Raji.Luc and CD19KOCD20 +CD22 +Raji.Luc cells . In vivo treatment efficacy and CAR-T persistence were also investigated in NCG murine model xenografted with Raji tumor line. 0.3x10 6 CAR +T cells or dose-matched untransduced T cells were given to NCG mice four days post i.v. implantation of Raji.Luc tumor cells. Tumor growth was monitored weekly by bioluminescence imaging until achieved endpoint (55 days), and CAR-T persistence was determined using genomic DNA level.Results: Tri-specific VHH CAR-T cells can mediate dose-dependent cytotoxicity against Raji tumor lines. Compared to mono- or bi-specific VHH or scFv CAR-T, tri-specific VHH CAR-T demonstrated equal or better cytolytic activity. Our lead tri-VHH CAR-T, LCAR-AIO, was able to specifically lyse K652 over-expressing single target such as CD19, CD20 or CD22, at the similar level to mono-specific CD19, CD20 or CD22 VHH CAR-T. Since no blocking effect of recognition against these three antigens was observed, our result suggested that all three VHHs in LCAR-AIO are functional. In comparison to anti-CD19 FMC63 scFv CAR-T, LCAR-AIO exhibited higher lytic activity and IFN-γ production against Raji.Luc tumor lines in vitro. In addition, LCAR-AIO retained its robust lytic activity and IFN-γ production when co-cultured with CD19KO-Raji.Luc cells while anti-CD19 FMC63 scFv CAR-T could not, suggesting LCAR-AIO may prevent tumor escape due to loss of CD19. Furthermore, comparison of LCAR-AIO to mono-scFv CAR-T (anti-CD19 FMC63-BBz, anti-CD20 Leu16-BBz or anti-CD22 m971-BBz) was performed in NCG mice xenografted with Raji cell line, LCAR-AIO exhibited better T cell expansion, longer persistence, and superior efficacy in eliminating tumors.Conclusions: Based on in vitro and in vivo preclinical data, tri-specific CD19xCD20xCD22 VHH CAR-T can be effective targeting tumors lack of CD19 expression, therefore, it has the potential of treating relapsed patients with prior CD19 CAR-T therapy. The feasibility of making tri-specific CAR-T would help to extend this technology to solid cancers where heterogeneity poses a major challenge at current stage. [Display omitted] DisclosuresZhou: Legend Biotech: Current Employment, Current equity holder in publicly-traded company. Han: Legend Biotech: Current Employment, Current equity holder in publicly-traded company. Pan: Legend Biotech: Current Employment, Current equity holder in publicly-traded company. Sang: Legend Biotech: Current Employment, Current equity holder in publicly-traded company. Shi: Legend Biotech: Current Employment. Feng: Legend Biotech: Current Employment. Xiao: Legend Biotech: Current Employment. Zhuang: Legend Biotech: Current Employment, Current equity holder in publicly-traded company. Wang: Legend Biotech: Current Employment, Current equity holder in publicly-traded company. Fan: Legend Biotech: Current Employment, Current equity holder in publicly-traded company.

Risk Factors and Protective Immunity Against Rabies in Unvaccinated Butchers Working at Dog Slaughterhouses in Northern Vietnam

Vietnam is a rabies-endemic country where eating dog meat is customary. However, the risks of rabies transmission to dog slaughtering and processing workers have not been identified. This study aimed to determine the rabies neutralizing antibody (NTA) and risk factors in dog slaughterers to propose appropriate intervention methods for this occupational group. In 2016, a cross-sectional study on NTA against rabies virus and related factors was conducted among 406 professional dog slaughterers in Vietnam. The participants were interviewed using a structured questionnaire, and their sera were tested for rabies NTA by a rapid focus fluorescence inhibition test. Statistical algorithms were used to analyze the data. The results showed that most of the professional dog butchers (344/406 subjects, 84.7%) had no rabies NTA. Interestingly, 7.8% (29/373) had NTA without a rabies vaccination history. Over 5 years of experience as a dog butcher was positively associated with the presence of NTA in unvaccinated individuals (OR = 6.16, P = 0.001). The NTA in vaccinated butchers was present in higher titer and for longer persistence to those of other previously reported professionals, which is possibly as a result of multiple exposures to low levels of rabies virus antigens during dog slaughtering. Our study demonstrated that professional dog butchers in Vietnam are at a high risk of rabies virus infection, apart from those with common bite experiences. In countries where dog meat consumption is customary, rabies control and prevention activities should focus on safety during dog trading and slaughtering.

PI3Kδ/γ inhibition promotes human CART cell epigenetic and metabolic reprogramming to enhance antitumor cytotoxicity

AbstractCurrent limitations in using chimeric antigen receptor T(CART) cells to treat patients with hematological cancers include limited expansion and persistence in vivo that contribute to cancer relapse. Patients with chronic lymphocytic leukemia (CLL) have terminally differentiated T cells with an exhausted phenotype and experience low complete response rates after autologous CART therapy. Because PI3K inhibitor therapy is associated with the development of T-cell–mediated autoimmunity, we studied the effects of inhibiting the PI3Kδ and PI3Kγ isoforms during the manufacture of CART cells prepared from patients with CLL. Dual PI3Kδ/γ inhibition normalized CD4/CD8 ratios and maximized the number of CD8+ T-stem cell memory, naive, and central memory T-cells with dose-dependent decreases in expression of the TIM-3 exhaustion marker. CART cells manufactured with duvelisib (Duv-CART cells) showed significantly increased in vitro cytotoxicity against CD19+ CLL targets caused by increased frequencies of CD8+ CART cells. Duv-CART cells had increased expression of the mitochondrial fusion protein MFN2, with an associated increase in the relative content of mitochondria. Duv-CART cells exhibited increased SIRT1 and TCF1/7 expression, which correlated with epigenetic reprograming of Duv-CART cells toward stem-like properties. After transfer to NOG mice engrafted with a human CLL cell line, Duv-CART cells expressing either a CD28 or 41BB costimulatory domain demonstrated significantly increased in vivo expansion of CD8+ CART cells, faster elimination of CLL, and longer persistence. Duv-CART cells significantly enhanced survival of CLL-bearing mice compared with conventionally manufactured CART cells. In summary, exposure of CART to a PI3Kδ/γ inhibitor during manufacturing enriched the CART product for CD8+ CART cells with stem-like qualities and enhanced efficacy in eliminating CLL in vivo.

Spatial and seasonal variations of PAHs in soil, air, and atmospheric bulk deposition along the plain to mountain transect in Hubei province, central China: Air-soil exchange and long-range atmospheric transport

Polycyclic aromatic hydrocarbons (PAHs) are a long-term environmental problem faced by human society. The sources of involuntary PAHs are complex, moreover, secondary emissions of fixed PAHs in the environment occur due to global change and disturbance of human activities. Samples of three environmental media including soil, air, and atmospheric bulk deposition were collected to observe the spatial distribution and seasonal variation, to discuss the source or sink of PAHs and their association with the air mass transport along the plain (Jianghan Plain, JHP) to mountain transect, and explore the geographic scope of the atmospheric transport influence. The results obtained showed that 16 individual PAHs generally existed in all environmental multimedia being studied, and the PAHs concentration in air, soil and deposition flux of atmospheric bulk was higher in JPH than in “Western Hubei Mountains” (WHMs). Considerably high PAHs concentrations were obtained from the soil, air and atmospheric bulk deposition in winter, summer, and both summer and winter, respectively. The air-soil fugacity fraction of PAHs indicated that the soil of Dajiuhu (DJH) is likely to be a sink. Backward air trajectory simulation confirmed that most of the air mass passes over the JHP before reaching DJH, combined with the (transport and persistence level III) TaPL3 model results JHP are acting as sources. However, seasonal changes lead to a shift in the roles of soil sources and sinks. The TaPL3 model calculated that PAHs are transported through water for a wider range of effects and a longer persistence, even up to 10 years.

Profiling CD8+ Tcell epitopes of COVID-19 convalescents reveals reduced cellular immune responses to SARS-CoV-2 variants.

Cellular immunity is important in determining the disease severity of COVID-19 patients. However, current understanding of SARS-CoV-2 epitopes mediating cellular immunity is limited. Here we apply T-Scan, a recently developed method, to identify CD8+ T cell epitopes from COVID-19 patients of four major HLA-A alleles. Several identified epitopes are conserved across human coronaviruses, which might mediate pre-existing cellular immunity to SARS-CoV-2. In addition, we identify and validate four epitopes that were mutated in the newly circulating variants, including the Delta variant. The mutations significantly reduce T cell responses to the epitope peptides in convalescent and vaccinated samples. We further determine the crystal structure of HLA-A∗02:01/HLA-A∗24:02 in complex with the epitope KIA_S/NYN_S, respectively, which reveals the importance of K417 and L452 of the spike protein for binding to HLA. Our data suggest that evading cellular immunity might contribute to the increased transmissibility and disease severity associated with the new SARS-CoV-2 variants.