Human mesenchymal stromal cells inhibit Mycobacterium avium replication in clinically relevant models of lung infection.

INTRODUCTION: Novel therapeutic strategies are urgently needed for Mycobacterium avium complex pulmonary disease (MAC-PD). Human mesenchymal stromal cells (MSCs) can directly inhibit MAC growth, but their effect on intracellular bacilli is unknown. We investigated the ability of human MSCs to reduce bacterial replication and inflammation in MAC-infected macrophages and in a murine model of MAC-PD. METHODS: Human monocyte-derived macrophages (MDMs) were infected with M. avium Chester strain and treated with human bone marrow-derived MSCs. Intracellular and extracellular colony-forming units (CFUs) were counted at 72 hours. Six-week-old female balb/c mice were infected by nebulisation of M. avium Chester. Mice were treated with 1×106 intravenous human MSCs or saline control at 21 and 28 days post-infection. Lungs, liver and spleen were harvested 42 days post-infection for bacterial counts. Cytokines were quantified by ELISA. RESULTS: MSCs reduced intracellular bacteria in MDMs over 72 hours (median 35% reduction, p=0.027). MSC treatment increased extracellular concentrations of prostaglandin E2 (PGE2) (median 10.1-fold rise, p=0.002) and reduced tumour necrosis factor-α (median 28% reduction, p=0.025). Blocking MSC PGE2 production by cyclo-oxygenase-2 (COX-2) inhibition with celecoxib abrogated the antimicrobial effect, while this was restored by adding exogenous PGE2. MSC-treated mice had lower pulmonary CFUs (median 18% reduction, p=0.012), but no significant change in spleen or liver CFUs compared with controls. CONCLUSION: MSCs can modulate inflammation and reduce intracellular M. avium growth in human macrophages via COX-2/PGE2 signalling and inhibit pulmonary bacterial replication in a murine model of chronic MAC-PD.

Taurolidine and Heparin as Catheter Lock Solution for Central Venous Catheters in Hemodialysis.

BACKGROUND: Chronic kidney disease can lead to end-stage renal disease, and the prevalence is increasing. Many patients starting hemodialysis require central venous catheters (CVCs). Catheter-related bloodstream infections (CRBSIs) are a common complication and lead to significant morbidity and mortality. Interventions to prevent CRBSI include antimicrobial lock therapy but concern for the development of antimicrobial resistance and adverse effects. Nonantimicrobial antiseptics as catheter lock solutions have also been used. Taurolidine and heparin catheter lock solution is first approved by the Food and Drug Administration for the prevention of CRBSI in patients on hemodialysis. Taurolidine has a unique mechanism of action and favorable safety profile. MECHANISM OF ACTION, PHARMACODYNAMICS, AND PHARMACOKINETICS: Taurolidine and heparin catheter lock solution have both antimicrobial and anticoagulant properties. Taurolidine is derivative of the amino acid taurine, and heparin is derived from porcine intestinal mucosa. Taurolidine not only damages microbial cell walls but also prevents the adherence of microorganisms to biological surfaces, preventing biofilm formation. Taurolidine and heparin catheter lock solution is intended to be used intraluminally within the catheter and should be aspirated. Because it is used locally, limited pharmacokinetic and pharmacodynamic data are available. CLINICAL TRIALS: The LOCK-IT-100 trial is a randomized, double-blind, phase 3 study, which included 795 end-stage renal disease patients on hemodialysis with CVC. Taurolidine and heparin was compared with the control heparin alone. The results of the study showed a 71% risk reduction in CRBSI for taurolidine and heparin arm (95% confident interval, 38%-86%, P = 0.0006). Other studies have also shown that taurolidine lock solution leads to decreased CRBSI episodes. Several systematic reviews and meta-analysis consisted of taurolidine in adult, and pediatric populations also showed reduction in the incidence of CRBSIs. THERAPEUTIC ADVANCE: Taurolidine and heparin lock solution represents a novel preventive strategy for those undergoing hemodialysis through a CVC by reducing the risk of CRBSI. This is significant progress because there are no other similar options available for patients for whom catheters are the only options for their life-saving treatment.

Enfacement illusions: Filling a knowledge gap in eating disorder risk assessment, prevention, and intervention?

Body image disturbance (BID) is central to eating disorders (EDs), yet the role of self-face perception has received limited empirical attention despite rising sociocultural pressures emphasizing facial appearance through technologies such as social media. Emerging evidence suggests impairments in self-face recognition accuracy and distorted perceptions of facial appearance among individuals with EDs. Enfacement illusions, involving the experimental induction of perceived ownership over another's face, offer a novel paradigm to comprehensively investigate the perceptual multisensory integration processes underlying self-face perception disturbances in ED populations. Such an approach may hold promise for elucidating core pathological mechanisms contributing to BID and ED psychopathology. We discuss how rigorous investigation of self-face perception through the enfacement illusion paradigm represents an innovative direction of research and/or clinical application that may advance etiological models of EDs and possibly inform interventions targeting the potentially multidimensional nature of body and facial image disturbances characterizing EDs. PUBLIC SIGNIFICANCE STATEMENT: Body image disturbance is central to eating disorders (EDs), yet, the role of face-related disturbances remains critically under-investigated. After summarizing findings on face-related disturbances in EDs we propose how enfacement illusions (i.e., the experimental induction of ownership over another's face) may elucidate self-face perception disturbances in EDs, and their underlying mechanisms. Enfacement illusions may also offer an intervention to potentially address multifaceted face and body image disturbances characterizing EDs.

Unraveling the Interplay: Soil Biogeochemical Factors Shaping the Efficacy of Anaerobic Soil Disinfestation in Suppressing Fusarium Root Rot of Strawberry.

Non-toxic alternatives to chemical soil fumigants for suppressing soilborne pathogens such as Fusarium oxysporum (Fo), one causative agent of strawberry black root rot complex prevalent in the southeastern U.S., are urgently needed. A promising alternative is anaerobic soil disinfestation (ASD), in which soil is amended with labile organic materials, irrigated to field capacity, and tarped to induce anaerobic fermentation for a brief period before planting. Pathogen-suppression mechanisms of ASD include anaerobic conditions and generation of reduced metal cations (Fe2+ and Mn2+) and volatile fatty acids (VFAs; e.g., acetic, n-butyric, isovaleric, and others). However, little is known about how the interaction between VFAs, reduced metals, soil texture, and liming influences suppression of Fo. We investigated Fo suppression by VFAs and reduced metal cations in both aqueous and soil-based incubation trials. Inoculum containing Fo chlamydospores was added to aqueous medium containing either 5 or 10 mmol/liter VFAs and either 0.01% or 0.05% (w/w) reduced metals. In soil-based incubations, chlamydospore-containing inoculum was applied to sandy, sandy loam, and silty clay soil saturated by solutions containing 10 or 20 mmol/liter VFAs with or without 0.05% (w/w) reduced metals. VFAs, particularly in combination with Fe2+ in aqueous solutions and Mn2+ in soils significantly reduced Fo viability. At the same time, liming and higher soil clay content reduced the effectiveness of VFAs and reduced metals for suppressing Fo, highlighting the influence of soil pH and soil texture on ASD effectiveness.

Control of backbone chemistry and chirality boost oligonucleotide splice switching activity.

Although recent regulatory approval of splice-switching oligonucleotides (SSOs) for the treatment of neuromuscular disease such as Duchenne muscular dystrophy has been an advance for the splice-switching field, current SSO chemistries have shown limited clinical benefit due to poor pharmacology. To overcome limitations of existing technologies, we engineered chimeric stereopure oligonucleotides with phosphorothioate (PS) and phosphoryl guanidine-containing (PN) backbones. We demonstrate that these chimeric stereopure oligonucleotides have markedly improved pharmacology and efficacy compared with PS-modified oligonucleotides, preventing premature death and improving median survival from 49 days to at least 280 days in a dystrophic mouse model with an aggressive phenotype. These data demonstrate that chemical optimization alone can profoundly impact oligonucleotide pharmacology and highlight the potential for continued innovation around the oligonucleotide backbone. More specifically, we conclude that chimeric stereopure oligonucleotides are a promising splice-switching modality with potential for the treatment of neuromuscular and other genetic diseases impacting difficult to reach tissues such as the skeletal muscle and heart.

First report of root rot of strawberry caused by Fusarium cugenangense, a member of the F. oxysporum species complex, in Tennessee, USA.

Strawberry (Fragaria × ananassa Duch) in Tennessee is cultivated on plastic mulched beds annually, and production is limited primarily by multiple oomycete and fungal root rot pathogens that result in reduced vigor and black root rot disease symptoms. In early June 2018, plants (cv. Chandler) with reduced shoot vigor and size, and black, necrotic stunted roots were collected from Rhea County, TN. Roots and crowns of 10 plants were cut into 1-3 cm pieces and surface sterilized with 0.6% NaOCl, followed by 70% ethanol for 1 min each, and plated on water agar. White mycelia produced after 3 days were transferred to potato dextrose agar amended with 10 mg/liter rifampicin. After 10 days, fungal colonies were light purple on the surface and dark purple on the colony underside, later developing blue-black pigmentation on the underside. Microconidia on carnation leaf agar were ovoid to ellipsoid, aseptate or septate and 8.0 to 24.2 (13.7) × 3.0 to 4.5 (3.8) µm in size, macroconidia were 3 to 5 septate and falcate to almost straight and 33.7 to 52.8 (44.4) × 4.0 to 5.5 (4.9) µm in size (n=80); both conidia were produced on monophialides. Chlamydospores were globose and subglobose, formed terminally and intercalary on aerial, submerged, and surface mycelium, singly or in pairs and were abundantly produced in sucrose broth and on synthetic nutrient-poor agar (SNA) (diam. 7.6 µm). Morphology was consistent with Fusarium oxysporum (Leslie and Summerell, 2006) and F. cugenangense, a member of the F. oxysporum species complex, as described by Maryani et al. (2019). Fungal mycelia were used for PCR (Phire Plant Direct PCR Master Mix, Thermo Scientific, CA) and the translational elongation factor 1-α (EF1α) region was amplified with primers EF-1/EF-2 (O'Donnell et al., 1998), internal transcribed spacer (ITS) regions amplified with primers ITS1/ITS2 (White et al. 1990), and the RNA polymerase second largest subunit region (RPB2) with primer pairs 5f2/7cr and 7cf/11ar (O'Donnell et al., 2022). PCR products of isolate SC5 were sequenced, and sequences compared to all sequences in the FUSARIOID-ID database using polyphasic identification (Crous et al., 2021) with EF1α (GenBank Accession No. ON703236) and RPB2 (OR472390) sequences. The highest similarity (100%) was with isolates of F. cugenangense, including ex-type isolate InaCC F984 (99.94% similarity) (Maryani et al., 2019). F. cugenangense is closely related to F. callistephi and F. elaeidis, but both species lack chlamydospores, and F. elaeidis has polyphialides (Lombard et al, 2019). To satisfy Koch's postulates, healthy rooted strawberry plants produced in soilless media were transplanted into 4 plastic pots (1.2-liter) containing 5% (w/v) fungal inoculum (grown on barley grain) and mixed into the top 5-cm of peat-based soilless medium. Pots were incubated at 25°C and 50% RH in a growth chamber. Four pots without inoculum served as controls. The trial was repeated. Within 8 weeks, all inoculated plants had low vigor, with necrotic and stunted roots. Root sections of control and inoculated plants were plated, and the pathogen was re-isolated from diseased roots of all inoculated plants only and confirmed as F. cugenangense based on morphology and sequence analysis. To our knowledge, this is the first report of F. cugenangense, or any member of the F. oxysporum species complex, causing root rot of strawberry in Tennessee and could be an important component of the production-limiting black root rot disease complex of strawberry.

Real-time control of urban drainage systems using neuro-evolution.

With climate change and urbanization, existing urban drainage systems are being stressed beyond their design capacity in many parts of the world. Real-time control (RTC) can improve the performance of these systems and reduce the need for system upgrades. However, developing optimal control policies for RTC is a challenging research area due to computational demands, high uncertainties and system dynamics. This study presents a new RTC method using neuro-evolution for controlling combined sewer overflow (CSO) in urban drainage systems. Neuro-evolution is an approach to neural network research by evolutionary algorithms. Neuro-evolution realizes RTC by training the control policy in advance, thus avoiding the online optimization process in the application period. The simulation results of the benchmark Astlingen network indicate that the trained control policy outperforms the equal filling degree strategy in terms of CSO volume reduction and robustness in the face of tank level uncertainty. The performance analysis of the typical CSO events shows that the control policy mainly makes positive contributions during 'small' CSO events rather than 'large' ones. In particular, the effectiveness of the control policy in 'small' CSO events is more prominent in the initial phase of the events compared with the final phase. This work stands to support a foundation for future studies in the control of urban water systems based on neuro-evolution.

The effect of green infrastructure on resilience performance in combined sewer systems under climate change.

Climate change is currently reshaping precipitation patterns, intensifying extremes, and altering runoff dynamics. Particularly susceptible to these impacts are combined sewer systems (CSS), which convey both stormwater and wastewater and can lead to combined sewer overflow (CSO) discharges during heavy rainfall. Green infrastructure (GI) can help mitigate these discharges and enhance system resilience under historical conditions; however, the quantification of its effect on resilience in a future climate remains unknown in the literature. This study employs a modified Global Resilience Analysis (GRA) framework for continuous simulation to quantify the impact of climate change on CSS resilience, particularly CSOs. The study assesses the efficacy of GI interventions (green roofs, permeable pavements, and bioretention cells) under diverse future rainfall scenarios based on EURO-CORDEX regional climate models (2085-2099) and three Representative Concentration Pathways (2.6, 4.5, 8.5 W/m2). The findings underscore a general decline in resilience indices across the future rainfall scenarios considered. Notably, the total yearly CSO discharge volume increases by a range of 145 % to 256 % in response to different rainfall scenarios. While GI proves effective in increasing resilience, it falls short of offsetting the impacts of climate change. Among the GI options assessed, green roofs routed to pervious areas exhibit the highest adaptive capacity, ranging from 9 % to 22 % at a system level, followed by permeable pavements with an adaptation capacity between 7 and 13 %. By linking the effects of future rainfall scenarios on CSO performance, this study contributes to understanding GI's potential as a strategic tool for enhancing urban resilience.

Can Blue-Green Infrastructure enhance resilience in urban drainage systems during failure conditions?

The need to enhance the resilience of urban drainage systems (UDSs) in view of emerging global climate change and urbanisation threats is well recognised. Blue-Green Infrastructure (BGI) provides a suitable strategy for building the resilience of existing UDSs. However, there are limited quantitative studies that provide evidence of their effectiveness for increased uptake in cities. In this research, coupled one-dimensional-two-dimensional (1D-2D) modelling is applied to assess the effectiveness of BGI that include rainwater harvesting systems, infiltration trenches, bioretention cells, and detention ponds using two case study UDSs located in Kampala that experience catastrophic pluvial flooding caused by extreme rainfall. The resulting flooding impacts are quantified considering 'failed' and 'non-failed' UDS initial states, using total flood volume and average flood duration as system performance indicators. The study results suggest that spatially distributed rainwater harvesting systems singularly lead to a reduction in total flood volume and average flood duration of 16-45% and 18-24% in the case study UDSs, respectively. Furthermore, the study results suggest that BGIs are more effective during moderate rainfall (T < 10 years). Based on the study findings, city scale implementation of multifunctional rainwater harvesting systems is recommended as a suitable strategy for enhancing UDSs' resilience.

Body Schema as Assessed by Upper Limb Left/Right Judgment Tasks Is Altered in Stroke: Implications for Motor Imagery Training.

BACKGROUND AND PURPOSE: Individuals with stroke often experience significant impairment of the upper limb. Rehabilitation interventions targeting the upper limb are typically associated with only small to moderate gains. The knowledge that body schema can be altered in other upper limb conditions has contributed to the development of tailored rehabilitation approaches. This study investigated whether individuals with stroke experienced alterations in body schema of the upper limb. If so, this knowledge may have implications for rehabilitation approaches such as motor imagery. METHODS: An observational study performed online consisting of left/right judgment tasks assessed by response time and accuracy of: (i) left/right direction recognition; (ii) left/right shoulder laterality recognition; (iii) left/right hand laterality recognition; (iv) mental rotation of nonembodied objects. Comparisons were made between individuals with and without stroke. Secondary comparisons were made in the stroke population according to side of stroke and side of pain if experienced. RESULTS: A total of 895 individuals (445 with stroke) participated. Individuals with stroke took longer for all tasks compared to those without stroke, and were less accurate in correctly identifying the laterality of shoulder (P < 0.001) and hand (P < 0.001) images, and the orientation of nonembodied objects (P < 0.001). Moreover, the differences observed in the hand and shoulder tasks were greater than what was observed for the control tasks of directional recognition and nonembodied mental rotation. No significant differences were found between left/right judgments of individuals with stroke according to stroke-affected side or side of pain. DISCUSSION AND CONCLUSIONS: Left/right judgments of upper limb are frequently impaired after stroke, providing evidence of alterations in body schema. The knowledge that body schemas are altered in individuals with longstanding stroke may assist in the development of optimal, well-accepted motor imagery programs for the upper limb.Video Abstract available for more insights from the authors (see the Video, Supplemental Digital Content 1, available at: http://links.lww.com/JNPT/A394).

First Report of Strawberry Black Root Rot Caused by Globisporangium sylvaticum in Tennessee, USA.

Globisporangium sylvaticum (syn. Pythium sylvaticum), is an oomycete that causes root rot and damping off of field crops, ornamentals, and vegetables. Several species in Pythiaceae are associated with black root rot of strawberry [(Fragaria × ananassa) Duchesne] (Millner 2006). Mature, stunted 'Chandler' strawberry plants, with reduced shoot vigor and black necrotic roots, were collected from Rhea County (June 2018) and Cumberland County, TN (May 2019). Aboveground symptoms occurred in low incidence (<5% of plants) in the fields. Plant roots were rinsed with tap water, cut into 1 to 3 cm pieces, and surface-disinfested (70% ethanol, 1 min) followed by a sterile water rinse. Root segments were crushed, placed on 20% V8 juice agar, and incubated in the dark at 21°C for 3 days. White fluffy mycelia grew from a majority of roots and coenocytic hyphae with globose hyphal swellings, delimited from hyphae by septa, were observed with microscopy. Hyphae were initially branched, curled, hyaline, and aseptate; however, septations were observed in older cultures. Globose structures (terminal and intercalary) were identified as sporangia [11 to 32 (avg. 22.1) µm diameter] when zoospores were observed (Parikh et al. 2022). Oospores [9 to 21 (avg. 16) µm diameter] were globose, smooth, aplerotic, and thick-walled. Oogonia, with or without one or more inflated antheridia, were observed when isolates were paired in culture, characteristics consistent with descriptions of Campbell and Hendrix (1967), Pratt and Green (1971), van der Plaats-Niterink (1981), and Uzuhashi et al. (2010). Genomic DNA was extracted (Extract-N-Amp™; Sigma-Aldrich, MO) for PCR amplification of internal transcribed spacer (ITS) regions of rDNA with primers ITS1/ITS4 (White et al. 1990); ITS and large subunit rRNA regions with primers UN-up18S42/UN-lo28S22 (Robideau et al. 2011); and cytochrome c oxidase subunit I (COI) mitochondrial DNA with primers OomCoxI-Levup/OomCoxI-Levlo (Robideau et al. 2011). Primers ITS1/ITS4 were used to amplify isolate TN (GenBank Accession MW386310, which had 100% homology with reference isolate MK326528). Primers UN-up18S42/UN-lo28S22 amplified isolates SAP18 and OO1 (Accessions MZ881935 and MZ881936, which had 99.8% homology with HQ665236), and COI primers amplified isolate SAP18 (Accession OK020192, which had 100% homology with GU071816 and KT692835). To satisfy Koch's postulates, inoculum of G. sylvaticum grown on autoclaved wheat seeds was added (5% w/v) to planting mix (1 peat:1 sand, v/v). Young, rooted strawberry plants were planted in 1.2-L pots with infested (n = 6) and control (no pathogen, n = 6) mixes, which was saturated with deionized water. Pots were covered with clear plastic for 48 h to maintain high humidity. Plants were grown in a greenhouse (24°C avg.) for 8 weeks. The disease assay was repeated. All plants in infested mix died, with black, necrotic roots. Plants in the control mix were healthy and well-established. The pathogen was reisolated from roots of all inoculated plants and confirmed to be G. sylvaticum based on morphology and molecular analyses. Root disease of strawberry caused by G. sylvaticum has been reported in the USA (Campbell and Hendrix 1967; Nemec and Sanders 1970; Pratt and Green 1971). This is the first report of G. sylvaticum causing root rot of strawberry in Tennessee. With the loss of methyl bromide, sustainable disease control strategies are needed to provide effective management options for strawberry black root rot.

Global resilience analysis of combined sewer systems under continuous hydrologic simulation.

Managing and reducing combined sewer overflow (CSO) discharges is crucial for enhancing the resilience of combined sewer systems (CSS). However, the absence of a standardised resilience analysis approach poses challenges in developing effective discharge reduction strategies. To address this, our study presents a top-down method that expands the existing Global Resilience Analysis to quantify resilience performance in CSS. This approach establishes a link between threats (e.g., rainfall) and impacts (e.g., CSOs) through continuous and long-term simulation, accommodating various rainfall patterns, including extreme events. We assess CSO discharge impacts from a resilience perspective by introducing eight new metrics. We conducted a case study in Fehraltorf, Switzerland, analysing the performance of three green infrastructure (GI) types (bioretention cells, green roofs, and permeable pavements) over 38 years. The results demonstrated that GI enhanced all resilience indices, with variations observed in individual CSO performance metrics and their system locations. Notably, in Fehraltorf, green roofs emerged as the most effective GI type for improving resilience, while the downstream outfall displayed the highest resilience enhancement. Overall, our proposed method enables a shift from event-based to continuous simulation analysis, providing a standardised approach for resilience assessment. This approach informs the development of strategies for CSO discharge reduction and the enhancement of CSS resilience.

Enhanced CNS transduction from AAV.PHP.eB infusion into the cisterna magna of older adult rats compared to AAV9.

The development of high efficiency, central nervous system (CNS) targeting AAV-based gene therapies is necessary to address challenges in both pre-clinical and clinical investigations. The engineered capsids, AAV.PHP.B and AAV.PHP.eB, show vastly improved blood-brain barrier penetration compared to their parent serotype, AAV9, but with variable effect depending on animal system, strain, and delivery route. As most characterizations of AAV.PHP variants have been performed in mice, it is currently unknown whether AAV.PHP variants improve CNS targeting when delivered intrathecally in rats. We evaluated the comparative transduction efficiencies of equititer doses (6 × 1011vg) of AAV.PHP.eB-CAG-GFP and AAV9-CAG-GFP when delivered into the cisterna magna of 6-9-month old rats. Using both quantitative and qualitative assessments, we observed consistently superior biodistribution of GFP+ cells and fibers in animals treated with AAV.PHP.eB compared to those treated with AAV9. Enhanced GFP signal was uniformly observed throughout rostrocaudal brain regions in AAV.PHP.eB-treated animals with matching GFP protein expression detected in the forebrain, midbrain, and cerebellum. Collectively, these data illustrate the benefit of intracisternal infusions of AAV.PHP.eB as an optimal system to distribute CNS gene therapies in preclinical investigations of rats, and may have important translational implications for the clinical CNS targeting.

HLA-DR polymorphism in SARS-CoV-2 infection and susceptibility to symptomatic COVID-19.

SARS-CoV-2 infection results in different outcomes ranging from asymptomatic infection to mild or severe disease and death. Reasons for this diversity of outcome include differences in challenge dose, age, gender, comorbidity and host genomic variation. Human leukocyte antigen (HLA) polymorphisms may influence immune response and disease outcome. We investigated the association of HLAII alleles with case definition symptomatic COVID-19, virus-specific antibody and T-cell immunity. A total of 1364 UK healthcare workers (HCWs) were recruited during the first UK SARS-CoV-2 wave and analysed longitudinally, encompassing regular PCR screening for infection, symptom reporting, imputation of HLAII genotype and analysis for antibody and T-cell responses to nucleoprotein (N) and spike (S). Of 272 (20%) HCW who seroconverted, the presence of HLA-DRB1*13:02 was associated with a 6·7-fold increased risk of case definition symptomatic COVID-19. In terms of immune responsiveness, HLA-DRB1*15:02 was associated with lower nucleocapsid T-cell responses. There was no association between DRB1 alleles and anti-spike antibody titres after two COVID vaccine doses. However, HLA DRB1*15:01 was associated with increased spike T-cell responses following both first and second dose vaccination. Trial registration: NCT04318314 and ISRCTN15677965.

Optimizing aminoglycoside selection for KPC-producing Klebsiella pneumoniae with the aminoglycoside-modifying enzyme (AME) gene aac(6')-Ib.

OBJECTIVES: KPC-producing Klebsiella pneumoniae (KPC-Kp) isolates commonly co-harbour the aminoglycoside-modifying enzyme (AME) gene aac(6')-Ib, which encodes an AME that can confer resistance to some of the commercially available aminoglycosides. We sought to determine the influence of AAC(6')-Ib in KPC-Kp on the pharmacodynamic activity of aminoglycosides. METHODS: Six KPC-Kp clinical isolates, three with and three without aac(6')-Ib, were analysed. Using these isolates, the bacterial killing of amikacin, gentamicin and tobramycin was assessed in static time-kill experiments. The pharmacodynamic activity of the aminoglycosides was then assessed in a dynamic one-compartment infection model over 72 h using simulated human pharmacokinetics of once-daily dosing with amikacin (15 mg/kg), gentamicin (5 mg/kg) and tobramycin (5 mg/kg). RESULTS: At clinically relevant aminoglycoside concentrations in time-kill experiments and the dynamic one-compartment model, gentamicin was more active than amikacin or tobramycin against the isolates harbouring aac(6')-Ib. Amikacin, gentamicin and tobramycin all showed progressively reduced bacterial killing with exposure to repeated doses against most isolates in the dynamic one-compartment model. MIC values were generally not a good predictor of gentamicin pharmacodynamic activity against KPC-Kp, but were more reliable for amikacin and tobramycin. CONCLUSIONS: Gentamicin may be preferred over amikacin or tobramycin for treatment of KPC-Kp infections. However, gentamicin MICs are not a consistent predictor of its pharmacodynamic activity and unexpected treatment failures are possible.

Anaerobic Soil Disinfestation Efficacy Against Fusarium oxysporum Is Affected by Soil Temperature, Amendment Type, Rate, and C:N Ratio.

A meta-analysis of anaerobic soil disinfestation (ASD) efficacy against Fusarium oxysporum and F. oxysporum f. sp. lycopersici was conducted emphasizing effects of environment and organic amendment characteristics and pot and field studies conducted on ASD amendment C:N ratio and soil temperature effects on F. oxysporum f. sp. lycopersici inoculum survival. In a pot study, two organic amendments, dry molasses-based or wheat bran-based, applied at 4 mg of C/g of soil, with 40:1, 30:1, 20:1, and 10:1 C:N ratios, were evaluated against F. oxysporum f. sp. lycopersici at 15 to 25°C. This study was followed by a pot study with temperature regimes of 15 to 25°C and 25 to 35°C and two C:N ratios (20:1 and 40:1), and a field study at 40:1, 30:1, 20:1, and 10:1 C:N ratios, a 30:1 C:N ratio at a lower C rate (2 mg of C/g of soil), and an anaerobic control. Soil temperature >25°C and more labile amendments increased ASD suppression of F. oxysporum/F. oxysporum f. sp. lycopersici in the meta-analysis. In pot studies, F. oxysporum f. sp. lycopersici survival was reduced for molasses-based mixtures at 20:1 and 30:1 C:N ratios compared with wheat bran-based mixtures but not compared with the anaerobic control. At 25 to 35°C, all ASD treatments suppressed F. oxysporum f. sp. lycopersici relative to controls. In the field, all ASD treatments reduced F. oxysporum f. sp. lycopersici survival compared with the anaerobic control, and 4 mg of C/g of soil amendment rates induced higher anaerobic conditions and higher F. oxysporum f. sp. lycopersici mortality compared with the 2 mg of C/g of soil rate. Although amendment C:N ratios from 10 to 40:1 were similarly suppressive of F. oxysporum, lower temperatures reduced ASD effectiveness against F. oxysporum/F. oxysporum f. sp. lycopersici and further work is warranted to enhance suppression at soil temperatures <25°C.

The EPIPHA-KNEE trial: Explaining Pain to target unhelpful pain beliefs to Increase PHysical Activity in KNEE osteoarthritis - a protocol for a multicentre, randomised controlled trial with clinical- and cost-effectiveness analysis.

BACKGROUND: Despite well-established benefits of physical activity for knee osteoarthritis (OA), nine of ten people with knee OA are inactive. People with knee OA who are inactive often believe that physical activity is dangerous, fearing that it will further damage their joint(s). Such unhelpful beliefs can negatively influence physical activity levels. We aim to evaluate the clinical- and cost-effectiveness of integrating physiotherapist-delivered pain science education (PSE), an evidence-based conceptual change intervention targeting unhelpful pain beliefs by increasing pain knowledge, with an individualised walking, strengthening, and general education program. METHODS: Two-arm, parallel-design, multicentre randomised controlled trial involving 198 people aged ≥50 years with painful knee OA who do not meet physical activity guideline recommendations or walk regularly for exercise. Both groups receive an individualised physiotherapist-led walking, strengthening, and OA/activity education program via 4x weekly in-person treatment sessions, followed by 4 weeks of at-home activities (weekly check-in via telehealth), with follow-up sessions at 3 months (telehealth) and 5 and 9 months (in-person). The EPIPHA-KNEE group also receives contemporary PSE about OA/pain and activity, embedded into all aspects of the intervention. Outcomes are assessed at baseline, 12 weeks, 6 and 12 months. Primary outcomes are physical activity level (step count; wrist-based accelerometry) and self-reported knee symptoms (WOMAC Total score) at 12 months. Secondary outcomes are quality of life, pain intensity, global rating of change, self-efficacy, pain catastrophising, depression, anxiety, stress, fear of movement, knee awareness, OA/activity conceptualisation, and self-regulated learning ability. Additional measures include adherence, adverse events, blinding success, COVID-19 impact on activity, intention to exercise, treatment expectancy/perceived credibility, implicit movement/environmental bias, implicit motor imagery, two-point discrimination, and pain sensitivity to activity. Cost-utility analysis of the EPIPHA-KNEE intervention will be undertaken, in addition to evaluation of cost-effectiveness in the context of primary trial outcomes. DISCUSSION: We will determine whether the integration of PSE into an individualised OA education, walking, and strengthening program is more effective than receiving the individualised program alone. Findings will inform the development and implementation of future delivery of PSE as part of best practice for people with knee OA. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry: ACTRN12620001041943 (13/10/2020).

Endophytic Beauveria bassiana increases galling of 'Rutgers' tomato roots with Meloidogyne incognita.

Beauveria bassiana is endophytic in many plant species and has been shown to protect host plants against insect pests and plant pathogens. However, less is known about its activity against plant-parasitic nematodes. In vitro and plant assays were conducted to determine the effect of B. bassiana 11-98 (Bb) on Meloidogyne incognita (root-knot nematode; RKN). Beauveria bassiana was confirmed as an endophyte in 'Rutgers' tomato and colonization patterns of Bb in 'Rutgers' (highly susceptible to RKN) were compared with those in 'Mountain Spring' (less susceptible to RKN). In greenhouse tests with 'Rutgers' at 30 and 60 days after treatment (DAT) with RKN and Bb, there were few differences in plant growth variables among treatments in repeated trials. However, RKN root galling and egg count/root system were enhanced in plants treated with Bb at 60 DAT. In an in vitro assay with egg masses from greenhouse tests, the percentages of hatched eggs, and mobile and immobile nematodes did not differ significantly for RKN and RKN+Bb treatments. The presence of viable Bb from roots was confirmed by collecting egg suspensions from root galls and plating them on selective medium. Colonies of Bb were verified on agar medium, but no parasitism of RKN eggs was observed. Research is needed to investigate factors responsible for increased galling by RKN in the presence of endophytic Bb in 'Rutgers' tomato.

Optimizing intracellular antibodies (intrabodies/nanobodies) to treat neurodegenerative disorders.

Intrabodies (both single-chain Fv and single-domain VH, VHH, and VL nanobodies) offer unique solutions to some of the challenges of delivery and target engagement posed by immunotherapeutics for the brain and other areas of the nervous system. The specificity, which includes the recognition of post-translational modifications, and capacity for engineering that characterize these antibody fragments can be especially well-focused when the genes encoding only the binding sites of the antibody are expressed intracellularly. Multifunctional constructs use fusions with peptides that can re-target antigen-antibody complexes to enhance both pharmacodynamic activity and intracellular solubility simultaneously. Fusions with proteolytic targeting signals, such as the PEST degron, greatly enhance potency in some cases. Stem cell transplants can be protected from exogenous misfolded proteins by stable transfection with intrabodies. Tandem expression to target two or more misfolding proteins in one treatment may be especially valuable for proteostatic disruptions due to genetic, aging, or toxic triggers. Advances in bioinformatics, screening protocols, and especially gene therapy are showing great promise for intrabody/ nanobody treatments of a full range of neurological disorders, including Alzheimer's disease and related tau dementias, Parkinson's disease and Lewy body diseases, Huntington's disease, amyotrophic lateral sclerosis, and prion diseases, among others.

Regulatory Implications of Integrated Real-Time Control Technology under Environmental Uncertainty.

Integrated real-time control (RTC) of urban wastewater systems, which can automatically adjust system operation to environmental changes, has been found in previous studies to be a cost-effective strategy to strike a balance between good surface water quality and low greenhouse gas emissions. However, its regulatory implications have not been examined. To investigate the effective regulation of wastewater systems with this technology, two permitting approaches are developed and assessed in this work: upstream-based permitting (i.e., environmental outcomes as a function of upstream conditions) and means-based permitting (i.e., prescription of an optimal RTC strategy). An analytical framework is proposed for permit development and assessment using a diverse set of high performing integrated RTC strategies and environmental scenarios (rainfall, river flow rate, and water quality). Results from a case study show that by applying means-based permitting, the best achievable, locally suitable environmental outcomes (subject to 10% deviation) are obtained in over 80% of testing scenarios (or all testing scenarios if 19% of performance deviation is allowed) regardless of the uncertain upstream conditions. Upstream-based permitting is less effective as it is difficult to set reasonable performance targets for a highly complex and stochastic environment.