Proteomic Ligandability Maps of Spirocycle Acrylamide Stereoprobes Identify Covalent ERCC3 Degraders.

Covalent chemistry coupled with activity-based protein profiling (ABPP) offers a versatile way to discover ligands for proteins in native biological systems. Here, we describe a set of stereo- and regiochemically defined spirocycle acrylamides and the analysis of these electrophilic "stereoprobes" in human cancer cells by cysteine-directed ABPP. Despite showing attenuated reactivity compared to structurally related azetidine acrylamide stereoprobes, the spirocycle acrylamides preferentially liganded specific cysteines on diverse protein classes. One compound termed ZL-12A promoted the degradation of the TFIIH helicase ERCC3. Interestingly, ZL-12A reacts with the same cysteine (C342) in ERCC3 as the natural product triptolide, which did not lead to ERCC3 degradation but instead causes collateral loss of RNA polymerases. ZL-12A and triptolide cross-antagonized one another's protein degradation profiles. Finally, we provide evidence that the antihypertension drug spironolactone─previously found to promote ERCC3 degradation through an enigmatic mechanism─also reacts with ERCC3_C342. Our findings thus describe monofunctional degraders of ERCC3 and highlight how covalent ligands targeting the same cysteine can produce strikingly different functional outcomes.

Identification of subfunctionalized aggregate-remodeling J-domain proteins in Arabidopsis thaliana.

J-domain proteins (JDPs) are critical components of the cellular protein quality control machinery, playing crucial roles in preventing the formation and, solubilization of cytotoxic protein aggregates. Bacteria, yeast, and plants additionally have large, multimeric heat shock protein 100 (Hsp100)-class disaggregases that resolubilize protein aggregates. JDPs interact with aggregated proteins and specify the aggregate-remodeling activities of Hsp70s and Hsp100s. However, the aggregate-remodeling properties of plant JDPs are not well understood. Here we identify eight orthologs of Sis1 (an evolutionarily conserved Class II JDP of budding yeast) in Arabidopsis thaliana with distinct aggregate-remodeling functionalities. Six of these JDPs associate with heat-induced protein aggregates in vivo and co-localize with Hsp101 at heat-induced protein aggregate centers. Consistent with a role in solubilizing cytotoxic protein aggregates, an atDjB3 mutant had defects in both solubilizing heat-induced aggregates and acquired thermotolerance as compared with wild-type seedlings. Next, we used yeast prions as protein aggregate models to show that the six JDPs have distinct aggregate-remodeling properties. Results presented in this study, as well as findings from phylogenetic analysis, demonstrate that plants harbor multiple, evolutionarily conserved JDPs with capacity to process a variety of protein aggregate conformers induced by heat and other stressors.

Hsp40/JDP Requirements for the Propagation of Synthetic Yeast Prions.

Yeast prions are protein-based transmissible elements, most of which are amyloids. The chaperone protein network in yeast is inexorably linked to the spreading of prions during cell division by fragmentation of amyloid prion aggregates. Specifically, the core "prion fragmentation machinery" includes the proteins Hsp104, Hsp70 and the Hsp40/J-domain protein (JDP) Sis1. Numerous novel amyloid-forming proteins have been created and examined in the yeast system and occasionally these amyloids are also capable of continuous Hsp104-dependent propagation in cell populations, forming synthetic prions. However, additional chaperone requirements, if any, have not been determined. Here, we report the first instances of a JDP-Hsp70 system requirement for the propagation of synthetic prions. We utilized constructs from a system of engineered prions with prion-forming domains (PrDs) consisting of a polyQ stretch interrupted by a single heterologous amino acid interspersed every fifth residue. These "polyQX" PrDs are fused to the MC domains of Sup35, creating chimeric proteins of which a subset forms synthetic prions in yeast. For four of these prions, we show that SIS1 repression causes prion loss in a manner consistent with Sis1's known role in prion fragmentation. PolyQX prions were sensitive to Sis1 expression levels to differing degrees, congruent with the variability observed among native prions. Our results expand the scope known Sis1 functionality, demonstrating that Sis1 acts on amyloids broadly, rather than through specific protein-protein interactions with individual yeast prion-forming proteins.

'It's a man's world': a gender-equitable scoping review of gender, transportation, and work.

The deeply embedded inequalities in gender which mark most contemporary societies have led to a world shaped by male perspectives. This world fails to accommodate adequately the needs and experiences of women: no more evident than in the transport sector, where a 'default male' perspective dominates the planning and policies that shape our roads, railways, airlines, and shipping. This paper argues that the ways in which masculinity infuses transport systems mean they are integral to debates on gender and work. They impact both the way women experience travel and their access to places of work. A multi-transport domain scoping study has been conducted to review the literature for key gender factors that influence the use of road, rail, aviation, and maritime transport modes. A multi-disciplinary approach is proposed which incorporates perspectives and methods from the social sciences that can help to foster Gender-Equitable Human Factors (GE-HF).Practitioner summary: This paper seeks to identify the gender issues related to transport and work. A scoping review provides key factors that detail how women are disadvantaged by current transport systems. It presents gaps in knowledge that future research needs to fill. Women must be included in key decisions within the transport sector.

A scoping review of youth and young adults' roles in natural disaster mitigation and response: considerations for youth wellbeing during a global ecological crisis.

BACKGROUND: Youth and young adults, ages 15-29, have been identified as assets in mitigation and management for natural disasters around the world. This study reviews the literature on disaster mitigation, response, and recovery following natural disasters with a focus on the engagement of youth and young adults. METHODS: The Arksey and O'Malley (2005) scoping review model was used to broadly summarize existing literature. A comprehensive electronic search was conducted using several databases. Keywords and index terms were searched using four concepts: (a) age (e.g., youth, young adult), (b) participation type (e.g., engagement, participation), (c) natural disaster type (e.g., hurricane, tsunami), and (d) disaster response type (e.g., planning, mitigation). RESULTS: The search yielded 214 articles published between 1990 and 2021. Of these articles, 19 met inclusion criteria. Identified literature focused on youth and young adults' mitigation and recovery efforts responding to natural disasters in several different countries. Themes of young people's participation or youth-led efforts focused on community mobilization and resilience (n = 5), youth empowerment and policy advocacy (n = 2), communication technology (n = 2), education-community partnerships (n = 9), and postdisaster rescue efforts (n = 1). CONCLUSIONS: With a changing climate and increased weather-related natural disasters, as well as in the context of a global health pandemic, community-based groups would benefit from learning from international efforts to engage young people in disaster mitigation and recovery efforts especially as it relates to promoting mental health and well-being.

The effect of intramuscular injection technique on injection associated pain; a systematic review and meta-analysis.

AIM: To review the effect of different intramuscular injection (IMI) techniques on injection associated pain, in adults. METHODS: The review protocol was registered on PROSPERO (CRD42019136097). MEDLINE, EMBASE, British Nursing Index and CINAHL were searched up to June 2020. Included studies were appraised and a meta-analysis, where appropriate, was conducted with a random effects model and test for heterogeneity. Standardised mean difference (SMD) with a 95% confidence interval in reported injection pain (intervention cf. control) was reported. RESULTS: 29 studies were included in the systematic review and 20 studies in the meta-analysis. 13 IMI techniques were identified. 10 studies applied local pressure to the injection site. Of these, applying manual pressure (4 studies, SMD = -0.85[-1.36,-0.33]) and Helfer (rhythmic) tapping (3 studies, SMD = -2.95[-5.51,-0.39]) to the injection site reduced injection pain, whereas the use of a plastic device to apply local pressure to the skin (ShotBlocker) did not significantly reduce pain (2 studies, SMD = -0.51[-1.58,0.56]). Acupressure techniques which mostly involved applying sustained pressure followed by intermittent pressure (tapping) to acupressure points local to the injection site reduced pain (4 studies: SMD = -1.62[-2.80,-0.44]), as did injections to the ventrogluteal site compared to the dorsogluteal site (2 studies, SMD = -0.43[-0.81,-0.06]). There was insufficient evidence on the benefits of the 'Z track technique' (2 studies, SMD = -0.20[-0.41,0.01]) and the cold needle technique (2 studies, SMD = -0.73[-1.83,0.37]) on injection pain. The effect of changing the needle after drawing up the injectate on injection pain was conflicting and warming the injectate did not reduce pain. Limitations included considerable heterogeneity, poor reporting of randomisation, and possible bias in outcome measures from unblinding of assessors or participants. CONCLUSIONS: Manual pressure or rhythmic tapping over the injection site and applying local pressure around the injection site reduced IMI pain. However, there was very high unexplained heterogeneity between studies and risk of significant bias within small studies.

nSeP: immune and metabolic biomarkers for early detection of neonatal sepsis-protocol for a prospective multicohort study.

INTRODUCTION: Diagnosing neonatal sepsis is heavily dependent on clinical phenotyping as culture-positive body fluid has poor sensitivity, and existing blood biomarkers have poor specificity.A combination of machine learning, statistical and deep pathway biology analyses led to the identification of a tripartite panel of biologically connected immune and metabolic markers that showed greater than 99% accuracy for detecting bacterial infection with 100% sensitivity. The cohort study described here is designed as a large-scale clinical validation of this previous work. METHODS AND ANALYSIS: This multicentre observational study will prospectively recruit a total of 1445 newborn infants (all gestations)-1084 with suspected early-or late-onset sepsis, and 361 controls-over 4 years. A small volume of whole blood will be collected from infants with suspected sepsis at the time of presentation. This sample will be used for integrated transcriptomic, lipidomic and targeted proteomics profiling. In addition, a subset of samples will be subjected to cellular phenotype and proteomic analyses. A second sample from the same patient will be collected at 24 hours, with an opportunistic sampling for stool culture. For control infants, only one set of blood and stool sample will be collected to coincide with clinical blood sampling. Along with detailed clinical information, blood and stool samples will be analysed and the information will be used to identify and validate the efficacy of immune-metabolic networks in the diagnosis of bacterial neonatal sepsis and to identify new host biomarkers for viral sepsis. ETHICS AND DISSEMINATION: The study has received research ethics committee approval from the Wales Research Ethics Committee 2 (reference 19/WA/0008) and operational approval from Health and Care Research Wales. Submission of study results for publication will involve making available all anonymised primary and processed data on public repository sites. TRIAL REGISTRATION NUMBER: NCT03777670.

Adverse effects of a single dose of gentamicin in adults: a systematic review.

AIMS: To systematically review the frequency and type of adverse events associated with a single dose of intravenous or intramuscular gentamicin in adults, for any indication, in studies where a comparator was available. METHODS: A review protocol was developed and registered (PROSPERO: CRD42013003229). Studies were eligible for review if they: recruited participants aged ≥16 years; used gentamicin intramuscularly or intravenously as a single one-off dose; compared gentamicin to another medication or placebo; and monitored adverse events. MEDLINE, EMBASE, Cochrane Library, trial registries, conference proceedings and other relevant databases were searched up to November 2016. Risk of bias was assessed on all included studies. RESULTS: In total, 15 522 records were identified. After removal of duplicates, screening of title/abstracts for relevance and independent selection of full texts by two reviewers, 36 studies were included. Across all the included studies, 24 107 participants received a single one-off dose of gentamicin (doses ranged from 1 mg kg-1 to 480 mg per dose). Acute kidney injury was described in 2520 participants receiving gentamicin. The large majority of cases were reversible. There were no cases of ototoxicity reported in patients receiving gentamicin. A meta-analysis was not performed due to study heterogeneity. CONCLUSIONS: A significant number of patients saw a transient rise in creatinine after a single dose of gentamicin at doses up to 480 mg. Persistent renal impairment and other adverse events were relatively rare.

From idiopathic diabetes insipidus to neurodegenerative Langerhans cell histiocytosis--an unusual presentation and progression of disease.

Diabetes insipidus (DI) is rare in childhood and has a wide-ranging aetiology including the involvement of uncontrolled proliferation of dendritic cells in the hypothalamic-pituitary axis, characteristic of Langerhans cell histiocytosis (LCH). DI may manifest as a sequela of multisystem LCH disease involving skin, bone, liver, spleen and lymph nodes. In very rare cases patients diagnosed with LCH exhibit neurodegenerative changes, such as severe ataxia, tremor, dysarthria and intellectual impairment. We report a 2 1/2-year-old boy who presented initially with apparent idiopathic DI, developed anterior pituitary hormone deficiency and progressive neurological deterioration secondary to neurodegenerative LCH.

Validation of bioelectrical impedance analysis in adolescents across different ethnic groups.

Adolescent obesity is difficult to assess in multi-ethnic populations using BMI, due to variability in the BMI-fatness relationship. We aimed to describe body composition (BC), and to validate leg-leg bioelectrical impedance analysis (BIA), in adolescents from different ethnic groups using deuterium (D(2)O) as the reference method. Measurements were made of weight, height, total body water (TBW), and BIA (TANITA TBF-300) in 110 white, 170 Asian, and 102 black adolescents aged 11-15 years. TBW was converted to lean mass (LM) using assumed hydration of lean tissue. General linear models were used to compare BC by D(2)O between the ethnic groups. BC values from D(2)O were compared with TANITA values, and used to generate ethnic-specific prediction equations in the whole sample, and also in equation-generation (group 1) and cross-validation (group 2) subsamples. Ethnic variability in BMI did not reflect variability in adiposity. Asians had less LM than white and black adolescents, and less fat mass (FM) than white girls. TANITA in-built equations did not predict BC accurately across ethnic groups, with significant bias in white and Asian males, and Asian and black females. The new equation generated from the entire sample removes ethnic-specific mean biases. The group 1 equation showed no significant bias in any ethnic group when tested in group 2. We found significant variability in BC between ethnic groups that was not reflected by BMI. Manufacturers' equations are unsuitable for predicting BC in multi-ethnic populations, and our new equations are recommended.