Three Pillars of Support for Orphan Drug Programs: Individual Valuations, Societal Valuations, and Anonymous Altruism.

OBJECTIVES: To identify and describe potential societal and individual sources of support for orphan drug programs. METHODS: The generalized risk-adjusted cost-effectiveness (GRACE) method shows that acute illness and disability severity increase individuals' willingness to pay (WTP) for health gains. We develop a social welfare function (SWF) that incorporates individuals' own values, combined with politically- or ethically determined weights. We introduce the concept of horizontal equity-that individuals in similar situations should be treated similarly-into the SWF. Finally, we introduce anonymous altruism into individuals' utility functions-the desire to help others, without knowing their identity. RESULTS: Combined with the empirical link between disease severity and rarity, GRACE demonstrates heightened WTP for health gains, leading rational individuals to support orphan drug programs, our first pillar of support. Adding horizontal equity to the SWF further increases societal support for orphan drug programs. Anonymous altruism, focusing most strongly on those in the most-dire circumstances, leads to altruistic support for those with severe disorders. Because innovators' economic incentives lead them to focus on larger markets, anonymous altruistic individuals to specifically support orphan drug programs. The presence of free-rider problems translates this into public program support. CONCLUSIONS: We identify supporting three pillars for orphan drug programs: (1) individuals' desire for treatments to treat rare disease that are often severe and life-threatening; (2) the concept of horizontal equity in our SWF: (3) anonymous altruism, the desire to people, even when unknown, in dire circumstances.

Sequence and structure analyses of lytic polysaccharide monooxygenases mined from metagenomic DNA of humus samples around white-rot fungi in Cuc Phuong tropical forest, Vietnam.

Background: White-rot fungi and bacteria communities are unique ecosystems with different types of symbiotic interactions occurring during wood decomposition, such as cooperation, mutualism, nutritional competition, and antagonism. The role of chitin-active lytic polysaccharide monooxygenases (LPMOs) in these symbiotic interactions is the subject of this study. Method: In this study, bioinformatics tools were used to analyze the sequence and structure of putative LPMOs mined by hidden Markov model (HMM) profiles from the bacterial metagenomic DNA database of collected humus samples around white-rot fungi in Cuc Phuong primary forest, Vietnam. Two genes encoding putative LPMOs were expressed in E. coli and purified for enzyme activity assay. Result: Thirty-one full-length proteins annotated as putative LPMOs according to HMM profiles were confirmed by amino acid sequence comparison. The comparison results showed that although the amino acid sequences of the proteins were very different, they shared nine conserved amino acids, including two histidine and one phenylalanine that characterize the H1-Hx-Yz motif of the active site of bacterial LPMOs. Structural analysis of these proteins revealed that they are multidomain proteins with different functions. Prediction of the catalytic domain 3-D structure of these putative LPMOs using Alphafold2 showed that their spatial structures were very similar in shape, although their protein sequences were very different. The results of testing the activity of proteins GL0247266 and GL0183513 show that they are chitin-active LPMOs. Prediction of the 3-D structures of these two LPMOs using Alphafold2 showed that GL0247266 had five functional domains, while GL0183513 had four functional domains, two of which that were similar to the GbpA_2 and GbpA_3 domains of protein GbpA of Vibrio cholerae bacteria. The GbpA_2 - GbpA_3 complex was also detected in 11 other proteins. Based on the structural characteristics of functional domains, it is possible to hypothesize the role of chitin-active GbpA-like LPMOs in the relationship between fungal and bacterial communities coexisting on decomposing trees in primary forests.

Glaucoma Detection through a Novel Hyperspectral Imaging Band Selection and Vision Transformer Integration.

Conventional diagnostic methods for glaucoma primarily rely on non-dynamic fundus images and often analyze features such as the optic cup-to-disc ratio and abnormalities in specific retinal locations like the macula and fovea. However, hyperspectral imaging techniques focus on detecting alterations in oxygen saturation within retinal vessels, offering a potentially more comprehensive approach to diagnosis. This study explores the diagnostic potential of hyperspectral imaging for glaucoma by introducing a novel hyperspectral imaging conversion technique. Digital fundus images are transformed into hyperspectral representations, allowing for a detailed analysis of spectral variations. Spectral regions exhibiting differences are identified through spectral analysis, and images are reconstructed from these specific regions. The Vision Transformer (ViT) algorithm is then employed for classification and comparison across selected spectral bands. Fundus images are used to identify differences in lesions, utilizing a dataset of 1291 images. This study evaluates the classification performance of models using various spectral bands, revealing that the 610-780 nm band outperforms others with an accuracy, precision, recall, F1-score, and AUC-ROC all approximately at 0.9007, indicating its superior effectiveness for the task. The RGB model also shows strong performance, while other bands exhibit lower recall and overall metrics. This research highlights the disparities between machine learning algorithms and traditional clinical approaches in fundus image analysis. The findings suggest that hyperspectral imaging, coupled with advanced computational techniques such as the ViT algorithm, could significantly enhance glaucoma diagnosis. This understanding offers insights into the potential transformation of glaucoma diagnostics through the integration of hyperspectral imaging and innovative computational methodologies.

Synergistic Effect of Venetoclax and Bendamustine in Early T-cell Precursor Acute Lymphoblastic Leukemia.

BACKGROUND/AIM: To date, therapeutic options for T-cell acute lymphoblastic leukemia (T-ALL) remain very limited. This study evaluated the efficacy of monotherapies and combination therapies including a selective BCL-2 inhibitor for T-ALL cell lines, namely Jurkat, CCRF-CEM, and Loucy. MATERIALS AND METHODS: Loucy is an early T-precursor ALL (ETP-ALL) cell line characterized by an immature phenotype, whereas Jurkat and CCRF-CEM are late T-cell progenitor ALL (LTP-ALL) cell lines. Monotherapy was conducted with venetoclax, cytarabine, bendamustine, or azacytidine, whereas combination therapy was performed with venetoclax plus cytarabine, venetoclax plus bendamustine, or venetoclax plus azacytidine. Cell viability assay was conducted after 48 h using Trypan blue and the 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS). Statistical analysis for evaluating synergistic interactions between anticancer drugs was performed by using the SynergyFinder Plus and drc R package. RESULTS: Adding venetoclax to cytarabine, bendamustine, or azacitidine achieved an additive effect, with Loewe synergic scores ranging from -10 to 10 in Jurkat and CCRF-CEM. Conversely, the combination of venetoclax and cytarabine displayed an additive effect (Loewe synergic score: 8.45 and 5.82 with MTS and Trypan blue assays, respectively), whereas venetoclax plus bendamustine or azacitidine exhibited a synergistic effect (Loewe synergic score >10 with MTS assay) in Loucy. Remarkably, the Bliss/Loewe score revealed that the combination of venetoclax and bendamustine was the most synergistic, yielding a score of 13.832±0.55. CONCLUSION: The combination of venetoclax and bendamustine demonstrated the greatest synergistic effect in suppressing ETP-ALL cell proliferation. Further studies are warranted to determine the mechanisms for the synergism between venetoclax and bendamustine in high-risk T-ALL.

The presence of broadly neutralizing anti-SARS-CoV-2 RBD antibodies elicited by primary series and booster dose of COVID-19 vaccine.

Antibody-mediated immunity plays a key role in protection against SARS-CoV-2. We characterized B-cell-derived anti-SARS-CoV-2 RBD antibody repertoires from vaccinated and infected individuals and elucidate the mechanism of action of broadly neutralizing antibodies and dissect antibodies at the epitope level. The breadth and clonality of anti-RBD B cell response varies among individuals. The majority of neutralizing antibody clones lose or exhibit reduced activities against Beta, Delta, and Omicron variants. Nevertheless, a portion of anti-RBD antibody clones that develops after a primary series or booster dose of COVID-19 vaccination exhibit broad neutralization against emerging Omicron BA.2, BA.4, BA.5, BQ.1.1, XBB.1.5 and XBB.1.16 variants. These broadly neutralizing antibodies share genetic features including a conserved usage of the IGHV3-53 and 3-9 genes and recognize three clustered epitopes of the RBD, including epitopes that partially overlap the classically defined set identified early in the pandemic. The Fab-RBD crystal and Fab-Spike complex structures corroborate the epitope grouping of antibodies and reveal the detailed binding mode of broadly neutralizing antibodies. Structure-guided mutagenesis improves binding and neutralization potency of antibody with Omicron variants via a single amino-substitution. Together, these results provide an immunological basis for partial protection against severe COVID-19 by the ancestral strain-based vaccine and indicate guidance for next generation monoclonal antibody development and vaccine design.

Synthesis of Nano Sulfur/Chitosan-Copper Complex and Its Nematicidal Effect against Meloidogyne incognita In Vitro and on Coffee Pots.

Sulfur is one of the inorganic elements used by plants to develop and produce phytoalexin to resist certain diseases. This study reported a method for preparing a material for plant disease resistance. Sulfur nanoparticles (SNPs) stabilized in the chitosan-Cu2+ (CS-Cu2+) complex were synthesized by hydrolysis of Na2S2O3 in an acidic medium. The obtained SNPs/CS-Cu2+ complex consisting of 0.32% S, 4% CS, and 0.7% Cu (w/v), contained SNPs with an average size of ~28 nm as measured by transmission electron microscopy images. The X-ray diffraction pattern of the SNPs/CSCu2+ complex showed that SNPs had orthorhombic crystal structures. Interaction between SNPs and the CS-Cu2+ complex was also investigated by ultraviolet-visible. Results in vitro nematicidal effect of materials against Meloidogyne incognita showed that SNPs/CS-Cu2+ complex was more effective in killing second-stage juveniles (J2) nematodes and inhibiting egg hatching than that of CS and CS-Cu2+ complex. The values of LC50 in killing J2 nematodes and EC50 in inhibiting egg hatching of SNPs/CS-Cu2+ complex were 75 and 51 mg/l, respectively. These values were lower than those of CS and the CS-Cu2+ complex. The test results on the nematicidal effect against M. incognita on coffee pots showed that the SNPs/CS-Cu2+ complex was 100% effective at a concentration of 150 mg/l. Therefore, the SNPs/CS-Cu2+ complex could be considered as a biochemical material with potential for agricultural applications to control root-knot nematodes.

Induction of Human Wharton's Jelly of Umbilical Cord Derived Mesenchymal Stem Cells to Be Chondrocytes and Transplantation in Guinea Pig Model with Spontaneous Osteoarthritis.

Osteoarthritis (OA) is a degenerative joint disease commonly found in elderly people and obese patients. Currently, OA treatments are determined based on their condition severity and a medical professional's advice. The aim of this study was to differentiate human Wharton's jelly-derived mesenchymal stem cells (hWJ-MSCs) into chondrocytes for transplantation in OA-suffering guinea pigs. hWJ-MSCs were isolated using the explant culture method, and then, their proliferation, phenotypes, and differentiation ability were evaluated. Subsequently, hWJ-MSCs-derived chondrocytes were induced and characterized based on immunofluorescent staining, qPCR, and immunoblotting techniques. Then, early-OA-suffering guinea pigs were injected with hyaluronic acid (HA) containing either MSCs or 14-day-old hWJ-MSCs-derived chondrocytes. Results showed that hWJ-MSCs-derived chondrocytes expressed specific markers of chondrocytes including Aggrecan, type II collagen, and type X collagen proteins and ß-catenin, Sox9, Runx2, Col2a1, Col10a1, and ACAN gene expression markers. Administration of HA plus hWJ-MSCs-derived chondrocytes (HA-CHON) produced a better recovery rate of degenerative cartilages than HA plus MSCs or only HA. Histological assessments demonstrated no significant difference in Mankin's scores of recovered cartilages between HA-CHON-treated guinea pigs and normal articular cartilage guinea pigs. Transplantation of hWJ-MSCs-derived chondrocytes was more effective than undifferentiated hWJ-MSCs or hyaluronic acid for OA treatment in guinea pigs. This study provides a promising treatment to be used in early OA patients to promote recovery and prevent disease progression to severe osteoarthritis.

Implementation of Telepharmacy to Provide Medication Consultation Service for Patients: A Case Study From Thu Duc City Hospital, Ho Chi Minh City.

Telepharmacy is receiving significant attention as an innovative approach. The objective of this study is to assess the needs and evaluate the impact of telepharmacy applications in drug consultations at Thu Duc City Hospital. We used a cross-sectional research design and conducted a survey with the participation of leaders of the Faculty of Pharmacy, clinical pharmacists, dispensing pharmacists, and patients or their caregivers who receive medication at the Pharmacy of Thu Duc Hospital. We deployed a telepharmacy application for consulting on drug use and surveyed the satisfaction of patients/family members with the telepharmacy model. 60.3% of survey subjects expressed a desire to receive drug use consultations through telepharmacy if the hospital were to offer this service. One hundred percent of the pharmacists at the pharmacy and the hospital's leadership believe that telepharmacy can address patient consultation needs and improve the current physical facilities in the dispensing area. Over 90% of telepharmacy users reported being satisfied or very satisfied with the service. Telepharmacy has garnered the attention of patients, their caregivers, and the medical staff at the Outpatient Pharmacy of Thu Duc Hospital. The majority of users are satisfied with the drug use consultation service provided by telepharmacy. By expanding the good results of Thu Duc Hospital to other hospitals, more patients across Vietnam can benefit from this innovative approach.

Water hyacinths retain river plastics.

Rivers represent one of the main conduits for the delivery of plastics to the sea, while also functioning as reservoirs for plastic retention. In tropical regions, rivers are exposed to both high levels of plastic pollution and invasion of water hyacinths. This aquatic plant forms dense patches at the river surface that drift due to winds and currents. Recent work suggests that water hyacinths play a crucial role in influencing plastic transport, by efficiently trapping the majority of surface plastic within their patches. However, a comprehensive understanding of the interaction between water hyacinths and plastics is still lacking. We hypothesize that the properties relevant to plastic transport change due to their trapping in water hyacinth patches. In particular, the length scale, defined as the characteristic size of the transported material, is a key property in understanding how materials move within rivers. Here, we show that water hyacinth patches trap on average 54%-77% of all observed surface plastics at the measurement site (Saigon river, Vietnam). Both temporally and spatially, we found that plastic and water hyacinth presence co-occur. The formation of plastic-plant aggregates carries significant implications for both clean-up and monitoring purposes, as these aggregates can be detected from space and need to be jointly removed. In addition, the length scale of trapped plastics (∼4.0 m) was found to be forty times larger than that of open water plastics (∼0.1 m). The implications of this increased length scale for plastic transport dynamics are yet to be fully understood, calling for further investigation into travel distances and trajectories. The effects of plastic trapping likely extend to other key properties of plastic-plant aggregates, such as effective buoyancy and mass. Given the prevalence of plant invasion and plastic pollution in rivers worldwide, this research offers valuable insights into the complex environmental challenges faced by numerous rivers.

iGATE analysis improves the interpretability of single-cell immune landscape of influenza infection.

Influenza poses a persistent health burden worldwide. To design equitable vaccines effective across all demographics, it is essential to better understand how host factors such as genetic background and aging affect the single-cell immune landscape of influenza infection. Cytometry by time-of-flight (CyTOF) represents a promising technique in this pursuit, but interpreting its large, high-dimensional data remains difficult. We have developed a new analytical approach, in silico gating annotating training elucidating (iGATE), based on probabilistic support vector machine classification. By rapidly and accurately "gating" tens of millions of cells in silico into user-defined types, iGATE enabled us to track 25 canonical immune cell types in mouse lung over the course of influenza infection. Applying iGATE to study effects of host genetic background, we show that the lower survival of C57BL/6 mice compared with BALB/c was associated with a more rapid accumulation of inflammatory cell types and decreased IL-10 expression. Furthermore, we demonstrate that the most prominent effect of aging is a defective T cell response, reducing survival of aged mice. Finally, iGATE reveals that the 25 canonical immune cell types exhibited differential influenza infection susceptibility and replication permissiveness in vivo, but neither property varied with host genotype or aging. The software is available at https://github.com/UmichWenLab/iGATE.

Enhanced photocatalytic efficiency of porous ZnO coral-like nanoplates for organic dye degradation.

ZnO nanomaterials have been extensively used as photocatalysts for the removal of pollutants in aqueous environments. This study explores the enhanced photocatalytic performance of porous ZnO coral-like nanoplates synthesized via a one-pot wet-chemical method and subsequent annealing treatment. Characterization through scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM), powder X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDX), Raman spectroscopy, photoluminescence (PL) spectroscopy, and Brunauer-Emmett-Teller (BET) measurements confirmed the nanoplates' porous structure, single-crystal structure, 100 nm thickness, and 80 nm pore size. These unique structural characteristics of the ZnO coral-like nanoplates enabled effective photodegradation of the organic dye rhodamine B (RhB) under visible light irradiation. Under simulated sunlight, the ZnO photocatalyst exhibited exceptional performance, achieving a 97.3% removal rate of RhB after 210 minutes of irradiation. The prepared ZnO photocatalyst also showed remarkable photostability and regeneration capability for RhB photodegradation with a decreased efficiency of less than 15% after eight testing cycles. The potential mechanism of the ZnO photocatalyst toward RhB degradation was also studied and is discussed in detail.

Orally ingestible medication utilizing layered double hydroxide nanoparticles strengthened alginate and hyaluronic acid-based hydrogel bead for bowel disease management.

In the treatment of bowel diseases such as ulcerative colitis through oral administration, an effective drug delivery system targeting the colon is crucial for enhancing efficacy and minimizing side effects of therapeutic agents. This study focuses on the development of a novel nanocomposite hydrogel bead comprising a synergistic blend of biological macromolecules, namely sodium alginate (ALG) and hyaluronic acid (HA), reinforced with layered double hydroxide nanoparticles (LDHs) for the oral delivery of dual therapeutics. The synthesized hydrogel bead exhibits significantly enhanced gel strength and controllable release of methylprednisolone (MP) and curcumin (CUR), serving as an anti-inflammatory drug and a mucosal healing agent, compared to native ALG or ALG/HA hydrogel beads without LDHs. The physicochemical properties of the synthesized LDHs and hydrogel beads were characterized using various techniques, including scanning electron microscopy, zeta potential measurement, transmission electron microscopy, X-ray diffraction, and energy-dispersive X-ray spectroscopy. In vitro release studies of MP and CUR under simulated gastrointestinal tract (GIT) conditions demonstrate the superior controlled release property of the nanocomposite hydrogel bead, particularly in minimizing premature drug release in the upper GIT environment while sustaining release of over 82 % of drugs in the colonic environment. Thus, the modularly engineered carrier designed for oral colon targeting holds promise as a potential candidate for the treatment of ulcerative colitis.

The genetic landscape of chromosomal aberrations in 3776 Vietnamese fetuses with clinical anomalies during pregnancy.

Background: Copy number variation sequencing (CNV-seq) is a powerful tool to discover structural genomic variation, but limitations associated with its retrospective study design and inadequate diversity of participants can be impractical for clinical application. Aim: This study aims to use CNV-seq to assess chromosomal aberrations in pregnant Vietnamese women. Materials & methods: A large-scale study was conducted on 3776 pregnant Vietnamese women with abnormal ultrasound findings. Results: Chromosomal aberrations were found in 448 (11.86%) women. Of these, 274 (7.26%) had chromosomal aneuploidies and 174 (4.61%) carried pathogenic/likely pathogenic CNVs. Correlations were established between chromosomal aberrations and various phenotypic markers. Conclusion: This comprehensive clinical study illuminates the pivotal role of CNV-seq in prenatal diagnosis for pregnancies featuring fetal ultrasound anomalies.

[Box: see text].

Microplastic contaminants detection in aquatic environment by hydrophobic cerium oxide nanoparticles.

Microplastics (MPs) poses a significant threat to ecosystems and human health, demanding immediate attention. The reported research work offers an effective and low cost method towards the detection of toxic MPs. In this study, hydrophobic cerium oxide nanoparticles (CeO2 NPs) are synthesized and applied as promising electrode material for the detection of two different types of MPs, i.e. polyethylene (PE) and polypropylene (PP). Through electrochemical analyses, such as cyclic voltammetry (CV) and linear sweep voltammetry (LSV), hydrophobic CeO2 NPs modified glassy carbon electrode (GCE) based sensor demonstrated remarkable sensitivity of ∼0.0343 AmLmg-1cm-2 and detection limit of ∼0.226 mgmL-1, with promising correlation coefficient (R2) towards the detection of PE (∼27-32 µm). Furthermore, hydrophobic CeO2 NPs modified GCE exhibited promising stability and reproducibility towards PE (∼27-32 µm), suggesting the promising potential of hydrophobic CeO2 NPs as electrode materials for an electrochemical microplastics detection.

In vitro and in silico docking and molecular dynamic of antimicrobial activities, alpha-glucosidase, and anti-inflammatory activity of compounds from the aerial parts of Mussaenda saigonensis.

Twelve compounds were isolated from Mussaenda saigonensis aerial parts through phytochemical analysis and the genus Mussaenda is the first place where the compounds 4-6 and 11-12 have been found. Based on the ability to inhibit NO production in RAW264.7 cells, compound 2 has demonstrated the strongest anti-inflammatory activity in vitro with an IC50 of 7.6 µM, as opposed to L-NMMA's IC50 of 41.3 µM. Compound 12 was found to be the most effective inhibitor of alpha-glucosidase enzyme in vitro, with an IC50 value of 42.4 µM (compared to 168 µM for acarbose). Compounds 1-12 were evaluated in vitro for antimicrobial activity using the paper dish method. Compound 11 demonstrated strong antifungal activity against M. gypseum with a MIC value of 50 µM. In silico docking for antimicrobial activity, pose 90 or compound 11 docked well to the 2VF5 enzyme, PDB, which explains why compound 11 had the highest activity in vitro. Entry 2/pose 280 demonstrated excellent anti-inflammatory activity in silico. The stability of the complex between pose 280 and the 4WCU enzyme for anti-inflammatory activity has been assessed using molecular dynamics over a simulation course ranging from 0 to 100 ns. It has been found to be stable from 60 and 100 ns. The Tyr 159 (95%, H-bond via water bridge), Asp 318 (200%, multiple contacts), Met 273 (75%, hydrophobic interaction via water bridge), and Gln 369 (75%, H-bond via water bridge) interacted well within the time range of 0 to 100 ns. It has more hydrophilic or polar pharmacokinetics.

Enhanced photocatalytic performance of ZnO under visible light by co-doping of Ta and C using hydrothermal method.

This study attempted to improve the photocatalytic activity of zinc oxide (ZnO) semiconductors in the visible light region by introducing the co-doping of carbon (C) and tantalum (Ta) to ZnO (ZTC) using a simple hydrothermal method with the respective precursors. The obtained uniform ZTC nanoparticles with an average crystal size of 29.30 nm (according to Scherrer's equation) revealed a redshift with a decrease in bandgap (Eg) from 3.04 eV to 2.88 eV, allowing the obtained photocatalyst to absorb the energy of the visible light for photocatalysis. Furthermore, the Zn 2p and Ta 4f core level spectra confirmed the presence of Zn2+ and Ta5+ in the ZTC sample. In addition, the infrared spectra identified hydrogen-related defects (HRDs), while the O 1s spectra indicated the existence of oxygen vacancies (VO). Electrochemical tests revealed improvement in the electron conductivity and charge separation of the obtained materials. To follow, the photocatalytic performance assessment was conducted by varying the C/Zn2+ ratios (5, 10, and 15 mol%) in ZTC samples, the initial RhB concentration (7, 15, and 30 ppm), and the pH of the RhB solution (3.0-10.0). The photodegradation on ZTC samples showed the most effectiveness for a 7 ppm RhB solution with a C/Zn2+ ratio of 10 mol% in the slightly alkaline medium (pH 9.0). Additionally, ZTC also exhibited commendable durability after being reused several times. The nature of RhB photodegradation was proposed and discussed via a mechanism at the end of this work.

Antibiotic prescribing practices of medical doctors in a resource-limited setting and the influence of individual perceptions and stewardship support: a survey in three tertiary hospitals in Vietnam.

Objectives: To understand antibiotic prescribing and influencing factors to inform antimicrobial stewardship (AMS) interventions to reduce unwanted consequences of antibiotic use in hospitals in Vietnam, a lower-middle-income country in Asia. Methods: We conducted a cross-sectional study of doctors at three tertiary hospitals using non-probability convenience sampling, through a paper-based (Hospitals 1 and 2) or electronic (Hospital 3) survey. Questions included items on perceptions regarding antibiotic resistance and AMS, prescribing practices, knowledge, demographics and training. We used principal components analysis and mixed-effects models to examine practices and identify influencing factors. Results: Among 314 surveyed participants, 61%, 57% and 59% in Hospitals 1, 2 and 3, respectively, felt certain about the appropriateness of their antibiotic prescriptions. In total, 9% reported sometimes prescribing antibiotics when not needed to meet patients' expectations, and 13% reported doing so to avoid perceived complications. Higher prescribing confidence was found among those with positive perceptions about AMS (P < 0.0001), whereas negative perceptions about colleagues' practices reduced this confidence (P < 0.0001). Individual preference for branded antibiotics was associated with more unnecessary prescribing whereas having higher prescribing confidence decreased the habits of prescribing when not needed. Conclusions: This study provides important implications for design of hospital interventions to address influencing factors on antibiotic prescribing in Vietnam and similar resource-limited settings. Specific interventions should target improving knowledge through education and training for doctors, enhancing the support from the AMS team, and promoting guidelines and policies for appropriate antibiotic use in hospital.

Plasma ctDNA liquid biopsy of IDH1, TERTp, and EGFRvIII mutations in glioma.

Background: Circulating tumor DNA has emerging clinical applications in several cancers; however, previous studies have shown low sensitivity in glioma. We investigated if 3 key glioma gene mutations IDH1, TERTp, and EGFRvIII could be reliably detected in plasma by droplet digital polymerase chain reaction (ddPCR) thereby demonstrating the potential of this technique for glioma liquid biopsy. Methods: We analyzed 110 glioma patients from our biobank with a total of 359 plasma samples (median 4 samples per patient). DNA was isolated from plasma and analyzed for IDH1, TERTp, and EGFRvIII mutations using ddPCR. Results: Total cfDNA was significantly associated with tumor grade, tumor volume, and both overall and progression-free survival for all gliomas as well as the grade 4 glioblastoma subgroup, but was not reliably associated with changes in tumor volume/progression during the patients' postoperative time course. IDH1 mutation was detected with 84% overall sensitivity across all plasma samples and 77% in the preoperative samples alone; however, IDH1 mutation plasma levels were not associated with tumor progression or survival. IDH1m plasma levels were not associated with pre- or postsurgery progression or survival. The TERTp C228T mutation was detected in the plasma ctDNA in 88% but the C250T variant in only 49% of samples. The EGFRvIII mutation was detected in plasma in 5 out of 7 patients (71%) with tissue EGFRvIII mutations in tumor tissue. Conclusions: Plasma ctDNA mutations detected with ddPCR provide excellent diagnostic sensitivity for IDH1, TERTp-C228T, and EGFRvIII mutations in glioma patients. Total cfDNA may also assist with prognostic information. Further studies are needed to validate these findings and the clinical role of ctDNA in glioma.

How tolerances, competition and dispersal shape benthic invertebrate colonisation in restored urban streams.

Biotic communities often respond poorly to river restoration activities and the drivers of community recovery after restoration are not fully understood. According to the Asymmetric Response Concept (ARC), dispersal capacity, species tolerances to stressors, and biotic interactions are three key drivers influencing community recovery of restored streams. However, the ARC remains to be tested. Here we used a dataset on benthic invertebrate communities of eleven restored stream sections in a former open sewer system that were sampled yearly over a period of eleven years. We applied four indices that reflect tolerance against chloride and organic pollution, the community's dispersal capacity and strength of competition to the benthic invertebrate taxa lists of each year and site. Subsequently, we used generalised linear mixed models to analyse the change of these indices over time since restoration. Dispersal capacity was high directly after restoration but continuously decreased over time. The initial communities thus consisted of good dispersers and were later joined by more slowly dispersing taxa. The tolerance to organic pollution also decreased over time, reflecting continuous improvement of water quality and an associated increase of sensitive species. On the contrary, chloride tolerances did not change, which could indicate a stable chloride level throughout the sampling period. Lastly, competition within the communities, reflected by interspecific trait niche overlap, increased with time since restoration. We show that recovery follows a specific pattern that is comparable between sites. Benthic communities change from tolerant, fast dispersing generalists to more sensitive, slowly dispersing specialists exposed to stronger competition. Our results lay support to the ARC (increasing role of competition, decreasing role of dispersal) but also underline that certain tolerances may still shape communities a decade after restoration. Disentangling the drivers of macroinvertebrate colonisation can help managers to better understand recovery trajectories and to define more realistic restoration targets.