Influence of pathogenic filaggrin variants on dupilumab treatment in atopic dermatitis

Background: Pathogenic variants in filaggrin (FLG) are associated with an increased risk of atopic dermatitis (AD). Objective: We evaluated the influence of FLG variants on the effectiveness of dupilumab treatment in AD. Methods: This prospective observational study included adult AD patients treated with dupilumab from the BioDay registry. FLG was analyzed with single-molecule molecular inversion probe–targeted sequencing. Novel mutations were confirmed by Sanger sequencing. Eczema Area and Severity Index (EASI), Investigator Global Assessment (IGA), numeric rating scale (NRS) pruritus, Dermatology Quality of Life Index (DLQI), and Patient-Oriented Eczema Measure (POEM) were assessed at baseline and at weeks 16 and 52. The study was registered at ClinicalTrials.gov as NCT03549416. Results: Genetic analysis of the 285 included patients showed biallelic pathogenic variants (FLG−/−) in 41 (14%), monoallelic pathogenic variants (FLG−/+) in 64 (23%), and wild-type alleles (FLG+/+) in 180 patients (63%). Three novel pathogenic variants were found. We observed no clinically relevant differences in EASI, IGA, NRS pruritus, DLQI, or total POEM scores for patients with and without pathogenic FLG variants at all time points. The FLG−/− group showed significantly higher POEM flaking and dryness scores at week 16 (P < .001 and P = .002, respectively) and week 52 (P < .001 and P = .016, respectively) compared to FLG+/+ as well as significant differences compared to FLG−/+, while differences in delta scores were nonsignificant. Conclusion: The effectiveness of dupilumab treatment in AD patients was not influenced by pathogenic FLG variants. However, patients with biallelic pathogenic FLG variants tended to have drier skin before and during dupilumab treatment compared to patients with monoallelic pathogenic variants or wild-type alleles

A reference human induced pluripotent stem cell line for large-scale collaborative studies.

Human induced pluripotent stem cell (iPSC) lines are a powerful tool for studying development and disease, but the considerable phenotypic variation between lines makes it challenging to replicate key findings and integrate data across research groups. To address this issue, we sub-cloned candidate human iPSC lines and deeply characterized their genetic properties using whole genome sequencing, their genomic stability upon CRISPR-Cas9-based gene editing, and their phenotypic properties including differentiation to commonly used cell types. These studies identified KOLF2.1J as an all-around well-performing iPSC line. We then shared KOLF2.1J with groups around the world who tested its performance in head-to-head comparisons with their own preferred iPSC lines across a diverse range of differentiation protocols and functional assays. On the strength of these findings, we have made KOLF2.1J and its gene-edited derivative clones readily accessible to promote the standardization required for large-scale collaborative science in the stem cell field

Duplex Ultrasound May Predict the Best Level of Lower Limb Amputation in Patients with Chronic Limb-Threatening Ischemia: A Retrospective Observational Cohort Study

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A mini-review of the anti-SARS-CoV-2 potency of Amaryllidaceae alkaloids

Abstract: Background Nature has perennially served as an infinite reservoir of diverse chemicals with numerous applications benefiting humankind. In recent years, due to the emerging COVID-19 pandemic, there has been a surge in studies on repurposing natural products as anti-SARS-CoV-2 agents, including plant-derived substances. Among all types of natural products, alkaloids remain one of the most important groups with various known medicinal values. The current investigation focuses on Amaryllidaceae alkaloids (AAs) since AAs have drawn significant scientific attention as anti-SARS-CoV-2 agents over the past few years. Purpose and study design This study serves as a mini-review, summarizing recent advances in studying the anti-SARS-CoV-2 potency of AAs, covering two aspects: structure-activity relationship and mechanism of action (MOA). Methods The study covers the period from 2019 to 2023. The information in this review were retrieved from common databases including Web of Science, ScienceDirect, PubMed and Google scholar. Reported anti-SARS-CoV-2 potency, cytotoxicity and possible biological targets of AAs were summarized and classified into different skeletal subclasses. Then, the structure-activity relationship (SAR) was explored, pinpointing the key pharmacophore-related structural moieties. To study the mechanism of action of anti-SARS-CoV-2 AAs, possible biological targets were discussed. Results In total, fourteen research articles about anti-SARS-CoV-2 was selected. From the SAR point of view, four skeletal subclasses of AAs (lycorine-, galanthamine-, crinine- and homolycorine-types) appear to be promising for further investigation as anti-SARS-CoV-2 agents despite experimental inconsistencies in determining in vitro half maximal inhibitory effective concentration (EC50). Narciclasine, haemanthamine- and montanine-type skeletons were cytotoxic and devoid of anti-SARS-CoV-2 activity. The lycorine-type scaffold was the most structurally diverse in this study and preliminary structure-activity relationships revealed the crucial role of ring C and substituents on rings A, C and D in its anti-SARS-CoV-2 activity. It also appears that two enantiomeric skeletons (haemanthamine- and crinine-types) displayed opposite activity/toxicity profiles regarding anti-SARS-CoV-2 activity. Pharmacophore-related moieties of the haemanthamine/crinine-type skeletons were the substituents on rings B, C and the dioxymethylene moiety. All galanthamine-type alkaloids in this study were devoid of cytotoxicity and it appears that varying substituents on rings C and D could enhance the anti-SARS-CoV-2 potency. Regarding MOAs, initial experimental results suggested Mpro and RdRp as possible viral targets. Dual functionality between anti-inflammatory activity on host cells and anti-SARS-CoV-2 activity on the SARS-CoV-2 virus of isoquinoline alkaloids, including AAs, were suggested as the possible MOAs to alleviate severe complications in COVID-19 patients. This dual functionality was proposed to be related to the p38 MAPK signaling pathway. Conclusion Overall, Amaryllidaceae alkaloids appear to be promising for further investigation as anti-SARS-CoV-2 agents. The skeletal subclasses holding the premise for further investigation are lycorine-, crinine-, galanthamine- and homolycorine-types

Towards a better understanding of the contribution of wastewater treatment plants to the microplastic pollution in receiving waterways

Microplastics (1 mu m-5 mm), are ubiquitous in daily-use products and regularly end up in the wastewater. The main part of the wastewater is treated in wastewater treatment plants (WWTPs), which allow for at least partial removal of microplastics. The present study aimed to understand the contribution of domestic wastewater to microplastic pollution in Flanders (Belgium) via two main discharge routes of microplastics: (1) the effluent, and (2) removed fractions. Furthermore the effect of effluent discharge on the microplastic contamination in the waterway was studied in both surface water and sediment samples of upstream and downstream locations of the discharge from three WWTPs. On average, 12.64 +/- 20.20 microplastic/L entered a WWTP (10 mu m-5 mm). The effluent contained on average 0.41 +/- 0.91 microplastic/L, resulting in an average removal efficiency of 97.46% +/- 2.33%, which is comparable with various (non-)European countries. Removal efficiencies are both polymer- and size-specific, and data suggest that smaller particles are less efficiently removed from the wastewater, which also causes an increased input of smaller particles to the environment. The sludge is the most efficient treatment process to remove microplastics. Despite the high removal efficiencies, still 1.11 x 10(7) +/- 3.07 x 10(7) microplastics end up in the nearby waterway daily. Nonetheless, based on the results gathered in the present study, this does not seem to impact the microplastic concentration in the waterway significantly. In summary, the present study offers a holistic approach in the research on the impact of wastewater on microplastic pollution in the ecosystem, integrating different discharge routes and measuring the impact on environmental microplastic pollution. Environ Toxicol Chem 2022;00:1-13. (c) 2022 SETA

Microplastic pollution in drinking water in Flanders

Recent studies described the occurrence of microplastics in nearly all environmental aquatic matrices. As expected, microplastics have also entered the food chain and can be ingested by humans via food and beverages such as salt, beer, drinking water and fruits. The purpose of this study was the identification of microplastics in large volumes of tap water that were derived from the purification of ground water, surface water or purified wastewater effluents in a densely populated region in western Europe, i.e. Flanders (Belgium). The samples were collected across Flanders at two different places in the supply chain: (1) purified water from drinking water treatment plants and (2) drinking water from conventional household taps. To identify potential microplastics, Fourier transform infrared microscopy was performed which enabled the identification of microplastic particles down to a size of 25 µm. A rescaling of the concentrations, based on a published method, was performed to enable the calculation of the microplastic concentration in the default microplastic range (1µm-5,000µm). An average of 0.02 ± 0.03 microplastics (>25µm) per liter (ranging between 0 and 0.06 microplastics per liter) was found in the samples taken in the water production centers. In three different water treatment plants – Essen, Egenhove and Gavers – no microplastic particles were found. Polypropylene is most commonly found polymer type after purification of the source water. Relating microplastic concentrations to the origin of the water (surface water, ground water and purified waste water), we observed that drinking water produced from groundwater did not contain microplastics in our samples. The drinking water from waste water effluents contains on average more microplastics (0.05 ± 0.02 MP/L) than that from surface water (0.02 ± 0.02 MP/L). However there is no significant difference in microplastic contamination according to the source of the water (p = 0.08). More data should be collected to confirm these results. It is not (yet) possible to assess potential risks of microplastics ingestion for human health as no epidemiological or other relevant studies on the effects of ingested microplastics have been published. More research is necessary to calculate a Derived No Effect Level (DNEL) for risk assessment of microplastic intake for human healt

Distribution of microplastics in freshwater systems in an urbanized region : a case study in Flanders (Belgium)

Microplastics (MPs) are an emerging pollutant of concern in all known aquatic ecosystems. However, studies at a regional scale on MP pollution in freshwater systems and the necessary risk assessments are limited. Therefore, in this study, we examined microplastic concentrations, size distributions, and polymer types in surface waters and sediments in the geographic region Flanders (Belgium), as a case study for a densely populated region and one of the most developed parts of Europe. Samples have been taken on nine different locations, of which five were repeated in a different weather condition. In total 43 aqueous and nine sediment samples have been collected. The quantity and identity of the microplastics in the samples were determined with μFTIR spectroscopy in the range of 25–1000 μm. The MPs' abundances in surface waters and sediments ranged from 0 to 4.8 MP L−1 (average = 0.48 MP L−1) and from 0 to 9558 MP kg−1 dry weight (average = 2774.57 ± 2317.93 MP kg−1 DW), respectively. Polystyrene and polypropylene were the most common polymer compositions found. No correlations were observed between microplastic concentrations in the sediment/the surface water samples and the measured environmental variables rainfall, conductivity, pH, dissolved oxygen content, waterway flow rate and width, and surrounding land use. Risk assessment results for the measured surface water concentrations through the risk quotient (RQ) method and the probabilistic risk assessment framework suggest that most of the sampled sites in Flanders posed negligible risks to freshwater biota, while this was not the case for some of the sediment concentrations. Our results illustrate the need to urgently develop analytical methods that can routinely measure the full size range of MP in environmental samples to adequately assess risks for the environment

Presence of microplastics in drinking water from different freshwater sources in Flanders (Belgium), an urbanized region in Europe

Abstract Microplastics (MPs) are emerging contaminants of concern in aquatic ecosystems. Up to now, only a few studies about MP contamination in drinking water have been published. In this study, we analysed drinking water originating from ground water, surface water and treated sewage water for the presence of MPs, collected in different drinking water treatment plants (DWTP, n = 9) and water taps (TW, n = 9) in the geographic region of Flanders (Belgium). We report measured microplastic concentrations, size distributions, and polymer types using μFTIR spectroscopy in the range of 25–1000 μm. The MPs’ abundances in the DWTP and TW samples were on average 0.02 ± 0.03 MPs L− 1 and 0.01 ± 0.02 MPs L− 1, respectively. We did not find significant differences comparing the obtained MP concentrations according to the origin of the water. Polypropylene (PP) and polyethylene terephthalate (PET) were the most common polymer types detected in the samples. Next, based on several theoretical assumptions, we extrapolated the measured MP concentrations in our samples to cover the full theoretical MP size range (1–5000 μm) to obtain estimates of the actual MP contamination levels. The rescaled particle concentrations (1 μm – 5000 mm) were on average 5.59 MPs L− 1 and 3.76 MPs L− 1 for the DWTP and TW samples, respectively. Based on a standard consumption of two liters of drinking water per day in combination with the measured concentration in this study, Flemish people consume 0.02 MPs per capita per day via drinking water. These findings contribute in our understanding of the microplastic pollution of drinking water, which is of concern due to the potential uptake of MPs in the human body

Dorsal unpaired median neurons of Locusta migratoria express ivermectin- and fipronil-sensitive glutamate-gated chloride channels

Together with type A GABA and strychnine-sensitive glycine receptors, glutamate-gated chloride channels ( GluCl) are members of the Cys-loop family of ionotropic receptors, which mediate fast inhibitory neurotransmission. To date, GluCls are found in invertebrates only and therefore represent potential specific targets for insecticides, such as ivermectin and fipronil. In this study, we identified the functional expression of GluCls in dorsal unpaired median ( DUM) neurons of the metathoracic ganglion of Locusta migratoria using electrophysiological and molecular biological techniques. In whole cell patch-clamped DUM neurons, glutamate-induced changes in both their membrane potentials (current-clamp) and currents (voltage-clamp) were dependent on the chloride equilibrium potential. On continuous application of glutamate, the glutamate-elicited current response became rapidly and completely desensitized. Application of glutamate in the presence of 10 mu M fipronil or 100 mu M picrotoxin reversibly decreased GluCl-mediated currents by 87 and 39%, respectively. Furthermore, 1 mu M ivermectin induced a persistent chloride current, suggesting the expression of ivermectin-sensitive GluCl alpha subunits. A degenerate PCR/RACE strategy was used to clone the full-length L. migratoria LmGlC1 alpha subunit. Finally, RT-PCR experiments demonstrated the presence of LmGluC1 alpha transcripts in locust DUM neurons. Our results provide the first direct evidence of a functional ivermectin-sensitive GluCl channel on the cell surface of DUM neurons of L. migratoria

Characterization of automatic treatment planning approaches in radiotherapy

Background and purpose: Automatic approaches are widely implemented to automate dose optimization in radiotherapy treatment planning. This study systematically investigates how to configure automatic planning in order to create the best possible plans. Materials and methods: Automatic plans were generated using protocol based automatic iterative optimization. Starting from a simple automation protocol which consisted of the constraints for targets and organs at risk (OAR), the performance of the automatic approach was evaluated in terms of target coverage, OAR sparing, conformity, beam complexity, and plan quality. More complex protocols were systematically explored to improve the quality of the automatic plans. The protocols could be improved by adding a dose goal on the outer 2 mm of the PTV, by setting goals on strategically chosen subparts of OARs, by adding goals for conformity, and by limiting the leaf motion. For prostate plans, development of an automated post-optimization procedure was required to achieve precise control over the dose distribution. Automatic and manually optimized plans were compared for 20 head and neck (H&N), 20 prostate, and 20 rectum cancer patients. Results: Based on simple automation protocols, the automatic optimizer was not always able to generate adequate treatment plans. For the improved final configurations for the three sites, the dose was lower in automatic plans compared to the manual plans in 12 out of 13 considered OARs. In blind tests, the automatic plans were preferred in 80% of cases. Conclusions: With adequate, advanced, protocols the automatic planning approach is able to create high-quality treatment plans