Prevention and Reduction of Anxiety in Autistic Preschoolers Through an Autism-Specific Parent-Mediated Intervention: A Pilot Randomised Controlled Trial Evaluating Short and Longer Term Outcomes

\ua9 The Author(s) 2024. Anxiety is a common co-occurring condition for autistic preschoolers. Whilst there has been extensive research evaluating anxiety prevention/reduction interventions for neurotypical preschoolers, such research is limited for autistic children. Fifty-seven parents of autistic 4–5-year olds, with varying levels of anxiety, participated in a randomised controlled trial of an autism-specific, parent-mediated intervention (CLK-CUES) to prevent or reduce anxiety in autistic preschoolers. Baseline, short-term (post-intervention) and longer term (12 months follow-up) assessments included child anxiety (ASC-ASD-P and PAS-R), intolerance of uncertainty, and parent well-being. (Trial registration ACTRN12620001322921). There were no reports of harmful effects of the intervention or trial. Linear mixed models show a significant group x time interaction for ASC-ASD-P Total score and the Uncertainty subscale with medium and large effect sizes. Post-hoc analyses show a significant decline for only the intervention group in anxiety (specifically, anxiety around uncertainty) from pre-post intervention, maintained at one year follow-up. There was no change on the PAS-R or other ASC-ASD-P subscales. CLK-CUES shows promise as a way to prevent and reduce anxiety in young autistic children, specifically anxiety related to uncertainty. Trials with larger samples are warranted. Findings also highlight the importance of using measures designed for autistic children

The analysis of acetaminophen (paracetamol) and seven metabolites in rat, pig and human plasma by U(H)PLC–MS

A U(H)PLC–MS/MS method is described for the analysis of acetaminophen and its sulphate, glucuronide, glutathione, cysteinyl and N-acetylcysteinyl metabolites in plasma using stable isotope-labeled internal standards. P-Aminophenol glucuronide and 3-methoxyacetaminophen were monitored and semi-quantified using external standards. The assay takes 7.5 min/sample, requires only 5 μl of plasma and involves minimal sample preparation. The method was validated for rat plasma and cross validated for human and pig plasma and mouse serum. LOQ in plasma for these analytes were 0.44 μg/ml (APAP-C), 0.58 μg/ml (APAP-SG), 0.84 μg/ml (APAP-NAC), 2.75 μg/ml (APAP-S), 3.00 μg/ml (APAP-G) and 16 μg/ml (APAP). Application of the method is illustrated by the analysis of plasma following oral administration of APAP to male Han Wistar rats

Metabolism and Effects on Endogenous Metabolism of Paracetamol (Acetaminophen) in a Porcine Model of Liver Failure

The metabolic fate, toxicity and effects on endogenous metabolism of paracetamol (acetaminophen, APAP) in 22 female Landrace cross large white pigs were evaluated in a model of acute liver failure (ALF). Anaesthetized pigs were initially dosed at 250 mg/kg via an oroduodenal tube with APAP serum concentrations maintained above 300 mg/L using maintenance doses of 0.5-4g/h until ALF. Studies were undertaken to determine both the metabolic fate of APAP and its effects on the endogenous metabolic phenotype of ALF in using 1H NMR spectroscopy. Increased concentrations of citrate combined with pre-ALF increases in circulating lactate, pyruvate and alanine in plasma suggest mitochondrial dysfunction and a switch in hepatic energy metabolism to glycolysis in response to APAP treatment. A specific liquid chromatography-tandem mass spectrometry assay was used to quantify APAP and metabolites. The major circulating and urinary metabolite of APAP was the phenolic glucuronide (APAP-G), followed by p-aminophenol glucuronide (PAP-G) formed from N-deacetylated APAP. The PAP produced by N-deacetylation was the likely cause of the methaemoglobinemia and kidney toxicity observed in this, and previous, studies in the pig. The phenolic sulfate of APAP, and the glutathione-derived metabolites of the drug were only found as minor components (with the cysteinyl conjugate detected but not the mercapturate). Given its low sulfation, combined with significant capacity for N-deacetylation the pig may represent a poor translational model for toxicology studies for compounds undergoing significant metabolism by sulfation, or which contain amide bonds which when hydrolysed to unmask an aniline lead to toxicity. However, the pig may provide a useful model where extensive amide hydrolysis is seen for drugs or environmental chemicals in humans, but not in e.g., the rat and dog which are the pre-clinical species normally employed for safety assessment

The analysis of acetaminophen (paracetamol) and seven metabolites in rat, pig and human plasma by U(H)PLC–MS

A U(H)PLC–MS/MS method is described for the analysis of acetaminophen and its sulphate, glucuronide, glutathione, cysteinyl and N-acetylcysteinyl metabolites in plasma using stable isotope-labeled internal standards. P-Aminophenol glucuronide and 3-methoxyacetaminophen were monitored and semi-quantified using external standards. The assay takes 7.5 min/sample, requires only 5 μl of plasma and involves minimal sample preparation. The method was validated for rat plasma and cross validated for human and pig plasma and mouse serum. LOQ in plasma for these analytes were 0.44 μg/ml (APAP-C), 0.58 μg/ml (APAP-SG), 0.84 μg/ml (APAP-NAC), 2.75 μg/ml (APAP-S), 3.00 μg/ml (APAP-G) and 16 μg/ml (APAP). Application of the method is illustrated by the analysis of plasma following oral administration of APAP to male Han Wistar rats

MerTK expressing hepatic macrophages promote the resolution of inflammation in acute liver failure

Objective: Acute liver failure (ALF) is characterised by overwhelming hepatocyte death and liver inflammation with massive infiltration of myeloid cells in necrotic areas. The mechanisms underlying resolution of acute hepatic inflammation are largely unknown. Here, we aimed to investigate the impact of Mer tyrosine kinase (MerTK) during ALF and also examine how the microenvironmental mediator, secretory leucocyte protease inhibitor (SLPI), governs this response. Design: Flow cytometry, immunohistochemistry, confocal imaging and gene expression analyses determined the phenotype, functional/transcriptomic profile and tissue topography of MerTK+ monocytes/macrophages in ALF, healthy and disease controls. The temporal evolution of macrophage MerTK expression and its impact on resolution was examined in APAP-induced acute liver injury using wild-type (WT) and Mer-deficient (Mer−/−) mice. SLPI effects on hepatic myeloid cells were determined in vitro and in vivo using APAP-treated WT mice. Results: We demonstrate a significant expansion of resolution-like MerTK+HLA-DRhigh cells in circulatory and tissue compartments of patients with ALF. Compared with WT mice which show an increase of MerTK+MHCIIhigh macrophages during the resolution phase in ALF, APAP-treated Mer−/− mice exhibit persistent liver injury and inflammation, characterised by a decreased proportion of resident Kupffer cells and increased number of neutrophils. Both in vitro and in APAP-treated mice, SLPI reprogrammes myeloid cells towards resolution responses through induction of a MerTK+HLA-DRhigh phenotype which promotes neutrophil apoptosis and their subsequent clearance. Conclusions: We identify a hepatoprotective, MerTK+, macrophage phenotype that evolves during the resolution phase following ALF and represents a novel immunotherapeutic target to promote resolution responses following acute liver injury

Use of 24kHz ultrasound to improve sulfate precipitation from wastewater

AbstractElevated sulfate concentrations in industrial effluent can lead to a number of significant problems, the most serious of which is the corrosion of concrete sewers as a result of hydrogen sulfide induced biogenic sulfuric acid attack; hydrogen sulfide can also create odor nuisance problems. The most common treatment process for sulfate removal from wastewaters is to precipitate it as gypsum using lime addition. Nevertheless, meeting discharge consent limits for sulfate can often present practical challenges due to the solubility of gypsum and so there is a need to investigate technological solutions that might provide for more consistent sulfate removal.This paper reports on the application of ultrasound during the sulfate precipitation process. We show that with as little as 10s sonication at 24kHz, significant increases in the rate of sulfate precipitation are observed. Particle size analysis, pH profiles and SEM micrographs, suggest that the likely mode of action is disaggregation of the calcium hydroxide particles, giving a greater solid–liquid interface, thus resulting in a faster dissolution rate and more readily available calcium ions. A range of experimental variables are studied, including the duration and power of sonication, as well as initial sulfate concentration and the effect of changing the time at which sonication is applied. For both sonicated and non-sonicated samples, precipitation commences almost immediately that the lime is added and so induction time is not an issue in this system

School anxiety, school attendance, and school “refusal”/school distress following an autism-specific parent-mediated intervention for anxiety in preschoolers

Shortreport: School anxiety, school attendance,and school “refusal”/school distressfollowing an autism-specific parent-mediated intervention for anxiety in preschoolersAutistic children have an increased likelihood of experiencing anxiety, even in the preschool years. Whilst there have been a number ofstudies evaluating interventions to prevent or reduce this anxiety in young autistic children, these have only evaluated the impact of such interventions using parent or child self-report data. This study expands upon previous workto evaluate whethera parent-mediated intervention for autistic preschool children’s anxiety has an impact beyond the home by evaluating children’sschool anxiety and school attendance post-transition into school. Teacher-reported data on school anxiety and parent-reported data on school attendance were compared for the intervention (n= 15) and control group (n= 13) from the CLK-CUES randomised controlled trial. Children whose parents received CLK-CUES not only had lower levels of separation anxiety in school but also had significantly lower non-attendance rates than the control group, and this was specifically associated with days missed due to school refusal/school distress. This indicates promise for autism-specific anxiety programs to reduce school anxiety and supportschool attendance in young autistic children