Simultaneous binding of Guidance Cues NET1 and RGM blocks extracellular NEO1 signaling.

During cell migration or differentiation, cell surface receptors are simultaneously exposed to different ligands. However, it is often unclear how these extracellular signals are integrated. Neogenin (NEO1) acts as an attractive guidance receptor when the Netrin-1 (NET1) ligand binds, but it mediates repulsion via repulsive guidance molecule (RGM) ligands. Here, we show that signal integration occurs through the formation of a ternary NEO1-NET1-RGM complex, which triggers reciprocal silencing of downstream signaling. Our NEO1-NET1-RGM structures reveal a "trimer-of-trimers" super-assembly, which exists in the cell membrane. Super-assembly formation results in inhibition of RGMA-NEO1-mediated growth cone collapse and RGMA- or NET1-NEO1-mediated neuron migration, by preventing formation of signaling-compatible RGM-NEO1 complexes and NET1-induced NEO1 ectodomain clustering. These results illustrate how simultaneous binding of ligands with opposing functions, to a single receptor, does not lead to competition for binding, but to formation of a super-complex that diminishes their functional outputs.

An HLA-E-targeted TCR bispecific molecule redirects T cell immunity against Mycobacterium tuberculosis.

Peptides presented by HLA-E, a molecule with very limited polymorphism, represent attractive targets for T cell receptor (TCR)-based immunotherapies to circumvent the limitations imposed by the high polymorphism of classical HLA genes in the human population. Here, we describe a TCR-based bispecific molecule that potently and selectively binds HLA-E in complex with a peptide encoded by the inhA gene of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis in humans. We reveal the biophysical and structural bases underpinning the potency and specificity of this molecule and demonstrate its ability to redirect polyclonal T cells to target HLA-E-expressing cells transduced with mycobacterial inhA as well as primary cells infected with virulent Mtb. Additionally, we demonstrate elimination of Mtb-infected cells and reduction of intracellular Mtb growth. Our study suggests an approach to enhance host T cell immunity against Mtb and provides proof of principle for an innovative TCR-based therapeutic strategy overcoming HLA polymorphism and therefore applicable to a broader patient population.

Structural basis of semaphorin-plexin cis interaction.

Semaphorin ligands interact with plexin receptors to contribute to functions in the development of myriad tissues including neurite guidance and synaptic organisation within the nervous system. Cell-attached semaphorins interact in trans with plexins on opposing cells, but also in cis on the same cell. The interplay between trans and cis interactions is crucial for the regulated development of complex neural circuitry, but the underlying molecular mechanisms are uncharacterised. We have discovered a distinct mode of interaction through which the Drosophila semaphorin Sema1b and mouse Sema6A mediate binding in cis to their cognate plexin receptors. Our high-resolution structural, biophysical and in vitro analyses demonstrate that monomeric semaphorins can mediate a distinctive plexin binding mode. These findings suggest the interplay between monomeric vs dimeric states has a hereto unappreciated role in semaphorin biology, providing a mechanism by which Sema6s may balance cis and trans functionalities.

Structure-guided stabilization of pathogen-derived peptide-HLA-E complexes using non-natural amino acids conserves native TCR recognition.

The nonpolymorphic class Ib molecule, HLA-E, primarily presents peptides from HLA class Ia leader peptides, providing an inhibitory signal to NK cells via CD94/NKG2 interactions. Although peptides of pathogenic origin can also be presented by HLA-E to T cells, the molecular basis underpinning their role in antigen surveillance is largely unknown. Here, we solved a co-complex crystal structure of a TCR with an HLA-E presented peptide (pHLA-E) from bacterial (Mycobacterium tuberculosis) origin, and the first TCR-pHLA-E complex with a noncanonically presented peptide from viral (HIV) origin. The structures provided a molecular foundation to develop a novel method to introduce cysteine traps using non-natural amino acid chemistry that stabilized pHLA-E complexes while maintaining native interface contacts between the TCRs and different pHLA-E complexes. These pHLA-E monomers could be used to isolate pHLA-E-specific T cells, with obvious utility for studying pHLA-E restricted T cells, and for the identification of putative therapeutic TCRs.

Adducing Knowledge Capabilities of Instrumental Techniques Through the Exploration of Heterostructures' Modification Methods.

The ongoing evolution of technology has facilitated the global research community to rapidly escalate the constant development of novel advancements in science. At the forefront of such achievements in the field of photocatalysis is the utilisation, and in oftentimes, the adaptation of modern instrumentation to understand photo-physical properties of complex heterostructures. For example, coupling in-situ X-ray Raman scattering spectroscopy for real-time degradation of catalytic materials.

TCRs with Distinct Specificity Profiles Use Different Binding Modes to Engage an Identical Peptide-HLA Complex.

The molecular rules driving TCR cross-reactivity are poorly understood and, consequently, it is unclear the extent to which TCRs targeting the same Ag recognize the same off-target peptides. We determined TCR-peptide-HLA crystal structures and, using a single-chain peptide-HLA phage library, we generated peptide specificity profiles for three newly identified human TCRs specific for the cancer testis Ag NY-ESO-1157-165-HLA-A2. Two TCRs engaged the same central peptide feature, although were more permissive at peripheral peptide positions and, accordingly, possessed partially overlapping peptide specificity profiles. The third TCR engaged a flipped peptide conformation, leading to the recognition of off-target peptides sharing little similarity with the cognate peptide. These data show that TCRs specific for a cognate peptide recognize discrete peptide repertoires and reconciles how an individual's limited TCR repertoire following negative selection in the thymus is able to recognize a vastly larger antigenic pool.

Do interventions principally targeting excessive alcohol use in young people improve depression symptoms?: a systematic review and meta-analysis.

BACKGROUND: Excessive alcohol use is common in young people and is associated with a range of adverse consequences including an increased risk of depression. Alcohol interventions are known to be effective in young people, however it is not known if these interventions can also improve depression. OBJECTIVE: To investigate whether psychosocial interventions principally targeting excessive alcohol use in young people reduce depression symptoms compared to controls. DESIGN: We conducted a systematic review and meta-analysis of controlled intervention trials, that measured depression symptoms at follow-up. We used a generic inverse variance random effect meta-analysis to pool the standardised mean difference in change in depression symptoms from baseline to follow-up between intervention and control arms. We used I2 to measure heterogeneity, the Cochrane tool for randomised trials to assess risk of bias, and Egger's tests to assess small study bias. DATA SOURCES: APA PsycNET, PubMed, Cochrane Central Register of Controlled Trials, Web of Science, Embase (including MEDLINE), and clinicaltrials.gov were searched for relevant studies published from inception to December 2020. Reference lists of studies were also searched, and authors contacted where articles presented insufficient data. STUDY ELIGIBILITY CRITERIA: Intervention studies that primarily targeted existing excessive alcohol use in young people (aged 10 to 24) and assessed depression outcomes at baseline with a minimum of four-week follow-up. RESULTS: Five studies were included in the meta-analysis. Interventions targeting excessive alcohol use were associated with a reduction in depression symptoms from baseline to follow-up when compared to control, standardised mean difference = - 0.26, and 95% confidence interval [- 0.41, - 0.12], p < .001. CONCLUSIONS: This study found evidence that interventions primarily targeting excessive alcohol use can reduce depression symptoms in young people. However, this finding should be taken with caution given concerns about risk of bias in all studies. More research is needed to examine whether these findings generalise beyond populations of undergraduate students primarily living in high income countries. TRIAL REGISTRATION: PROSPERO registration number: CRD42020177260 .

Solution Conformations of Curcumin in DMSO.

NAMFIS (NMR Analysis of Molecular Flexibility In Solution) has been applied to curcumin dissolved in DMSO. Quantitative 1H-1H distance constraints reduce a pool of candidate conformations to a solution collection of four enol conformations-two of these match curcumin crystallized with human transthyretin, and one is closely related to a single-crystal structure of curcumin.

Structural basis of semaphorin-plexin signalling.

Cell-cell signalling of semaphorin ligands through interaction with plexin receptors is important for the homeostasis and morphogenesis of many tissues and is widely studied for its role in neural connectivity, cancer, cell migration and immune responses. SEMA4D and Sema6A exemplify two diverse vertebrate, membrane-spanning semaphorin classes (4 and 6) that are capable of direct signalling through members of the two largest plexin classes, B and A, respectively. In the absence of any structural information on the plexin ectodomain or its interaction with semaphorins the extracellular specificity and mechanism controlling plexin signalling has remained unresolved. Here we present crystal structures of cognate complexes of the semaphorin-binding regions of plexins B1 and A2 with semaphorin ectodomains (human PLXNB1(1-2)-SEMA4D(ecto) and murine PlxnA2(1-4)-Sema6A(ecto)), plus unliganded structures of PlxnA2(1-4) and Sema6A(ecto). These structures, together with biophysical and cellular assays of wild-type and mutant proteins, reveal that semaphorin dimers independently bind two plexin molecules and that signalling is critically dependent on the avidity of the resulting bivalent 2:2 complex (monomeric semaphorin binds plexin but fails to trigger signalling). In combination, our data favour a cell-cell signalling mechanism involving semaphorin-stabilized plexin dimerization, possibly followed by clustering, which is consistent with previous functional data. Furthermore, the shared generic architecture of the complexes, formed through conserved contacts of the amino-terminal seven-bladed ß-propeller (sema) domains of both semaphorin and plexin, suggests that a common mode of interaction triggers all semaphorin-plexin based signalling, while distinct insertions within or between blades of the sema domains determine binding specificity.

A randomised controlled trial comparing opt-in and opt-out home visits for tracing lost participants in a prospective birth cohort study.

BACKGROUND: Attrition is an important problem in cohort studies. Tracing cohort members who have moved or otherwise lost contact with the study is vital. There is some debate about the acceptability and relative effectiveness of opt-in versus opt-out methods of contacting cohort members to re-engage them in this context. We conducted a randomised controlled trial to compare the two approaches in terms of effectiveness (tracing to confirm address and consenting to continue in the study), cost-effectiveness and acceptability. METHODS: Participants in this trial were individuals (young people and mothers) recruited to the Avon Longitudinal Study of Parents and Children (ALSPAC), who had not engaged with the study in the previous 5 years and for whom mail had been returned from their last known address. The sampling frame was restricted to those for whom database searching led to a potential new address being found in the Bristol area. 300 participants were randomly selected and assigned using stratified randomisation to the opt-in or opt-out arm. A tailored letter was sent to the potential new address, either asking participants to opt in to a home visit, or giving them the option to opt out of a home visit. Fieldworkers from Ipsos MORI conducted home visits to confirm address details. RESULTS: The proportion who were traced was higher in the opt-out arm (77/150 = 51 %) than the opt-in arm (6/150 = 4 %), as was the proportion who consented to continue in ALSPAC (46/150 = 31 % v 4/150 = 3 %). The mean cost per participant was £8.14 in the opt-in arm and £71.93 in the opt-out arm. There was no evidence of a difference in acceptability between the opt-in and opt-out approaches. CONCLUSION: Since the opt-in approach yielded very low response rates, and there were no differences in terms of acceptability, we conclude that the opt-out approach is the most effective method of tracing disengaged study members. The gains made in contacting participants must be weighed against the increase in cost using this methodology.

The 'Ireland' one-pot alcohol oxidation coupling reactions: celebrating 30 years of diverse synthesis.

In 1985, Robert Ireland and co-worker were devising synthetic routes to polyether ionophore antibiotics and during this process several highly reactive aldehydes were encountered, which made their isolation and subsequent elaboration difficult. To circumvent this problem, the synthetic route commenced from the alcohol which was oxidized to the aldehyde and, to the crude mixture, was added the nucleophile. This procedure, dubbed the 'Ireland' oxidative process was found to be highly successful and adapted by many research groups the world over. As we celebrate 30 years since this landmark discovery, the following review will take a journey of the most innovative and wonderful applications of this process.

Palladium-catalysed cross-coupling reaction of ultra-stabilised 2-aryl-1,3-dihydro-1H-benzo[d]1,3,2-diazaborole compounds with aryl bromides: A direct protocol for the preparation of unsymmetrical biaryls.

There has been a significant interest in organoboron compounds such as arylboronic acids, arylboronate esters and potassium aryltrifluoroborate salts because they are versatile coupling partners in metal-catalysed cross-coupling reactions. On the other hand, their nitrogen analogues, namely, 1,3,2-benzodiazaborole-type compounds have been studied extensively for their intriguing absorption and fluorescence characteristics. Here we describe the first palladium-catalysed Suzuki-Miyaura cross-coupling reaction of easily accessible and ultra-stabilised 2-aryl-1,3-dihydro-1H-benzo[d]1,3,2-diazaborole derivatives with various aryl bromides. Aryl bromides bearing electron-withdrawing, electron-neutral and electron-donating substituents are reacted under the catalytic system furnishing unsymmetrical biaryl products in isolated yields of up to 96% in only 10 minutes.

Attention-deficit hyperactivity disorder traits are a more important predictor of internalising problems than autistic traits.

Autism Spectrum Disorder (ASD) and Attention-Deficit Hyperactivity Disorder (ADHD) are both linked to internalising problems like anxiety and depression. ASD and ADHD also often co-occur, making their individual statistical contributions to internalising disorders difficult to investigate. To address this issue, we explored the unique associations of self-reported ASD traits and ADHD traits with internalising problems using a large general population sample of adults from the United Kingdom (N = 504, 49% male). Classical regression analyses indicated that both ASD traits and ADHD traits were uniquely associated with internalising problems. Dominance and Bayesian analyses confirmed that ADHD traits were a stronger, more important predictor of internalising problems. However, brief depression and anxiety measures may not provide a comprehensive index of internalising problems. Additionally, we focused on recruiting a sample that was representative of the UK population according to age and sex, but not ethnicity, a variable that may be linked to internalising disorders. Nevertheless, our findings indicate that while ASD and ADHD uniquely predict internalising problems, ADHD traits are a more important statistical predictor than ASD traits. We discuss potential mechanisms underlying this pattern of results and the implications for research and clinical practice concerning neurodevelopmental conditions.

Promiscuous recognition of MR1 drives self-reactive mucosal-associated invariant T cell responses.

Mucosal-associated invariant T (MAIT) cells use canonical semi-invariant T cell receptors (TCR) to recognize microbial riboflavin precursors displayed by the antigen-presenting molecule MR1. The extent of MAIT TCR crossreactivity toward physiological, microbially unrelated antigens remains underexplored. We describe MAIT TCRs endowed with MR1-dependent reactivity to tumor and healthy cells in the absence of microbial metabolites. MAIT cells bearing TCRs crossreactive toward self are rare but commonly found within healthy donors and display T-helper-like functions in vitro. Experiments with MR1-tetramers loaded with distinct ligands revealed significant crossreactivity among MAIT TCRs both ex vivo and upon in vitro expansion. A canonical MAIT TCR was selected on the basis of extremely promiscuous MR1 recognition. Structural and molecular dynamic analyses associated promiscuity to unique TCRß-chain features that were enriched within self-reactive MAIT cells of healthy individuals. Thus, self-reactive recognition of MR1 represents a functionally relevant indication of MAIT TCR crossreactivity, suggesting a potentially broader role of MAIT cells in immune homeostasis and diseases, beyond microbial immunosurveillance.

2-(3-Meth-oxy-phen-yl)-1,3-dihydro-1,3,2-benzodiaza-borole.

The title compound, C(13)H(13)BN(2)O, is one in a series of 1,3,2-benzodiaza-boroles featuring a 2-meth-oxy-phenyl substitution at the 2-position in the nitro-gen-boron heterocyle. The dihedral angle between the mean planes of the benzodiaza-borole and 2-meth-oxy-phenyl ring systems is 21.5 (1)°. There is an inter-molecular hydrogen bond between one of the NH groups and the meth-oxy O atom. This hydrogen bond leads to an infinite hydrogen-bonded chain colinear with the a axis.

Heterodinuclear Ru-Pt Complexes Bridged with 2,3-Bis(pyridyl)pyrazinyl Ligands: Studies on Kinetics, Deoxyribonucleic Acid/Bovine Serum Albumin Binding and Cleavage, In Vitro Cytotoxicity, and In Vivo Toxicity on Zebrafish Embryo Activities.

Di- and poly-homo/heteronuclear complexes have great potential as anticancer drugs. Here, we report their reactivity, deoxyribonucleic acid (DNA)/bovine serum albumin (BSA) binding and cleavage interactions, in vitro cytotoxicity, and in vivo zebrafish embryo toxicity of [(phen)2Ru(µ-L)PtCl2]2+ (phen = 1,10-phenanthroline and L = 2,3-bis(2-pyridyl)pyrazine, bpp, C1 ; 2,3-bis(2-pyridyl)quinoxaline, bpq, C2ial ; 2,3-bis(2-pyridyl)benzo[g]quinoxaline, bbq, C3 ) anticancer prodrugs. The substitution reactivity increases from C1 to C3 owing to an increase in the π-conjugation on the bridging chelate which facilitates π-back bonding. As a result, the electrophilicity index on the C3 complex increases than that on the complex C2 followed by C1 which leads to higher rates of substitution and thus the reactivity order follows C1 < C2 < C3 . The coordination of Ru at one end of each of the complexes enhances water solubility. Moreover, the charge addition of the two metal ions increases their reactivity toward substitution in addition to ensuring electrostatic interactions at target sites such as the DNA/BSA. Spectroscopic (UV-vis absorption and fluorescence quenching) titration and viscosity measurement results of the interactions of C1/2/3 with CT-DNA established the formation of stable, nonconvent C1/2/3 -DNA adducts with DNA most likely via the intercalative binding mode. Furthermore, studies with BSA showed a good binding affinity of these complexes owing to hydrophobic interactions with the coordinated ligands. The interactions of these complexes with DNA/BSA are in line with the reactivity trend, and all these experimental findings were further supported by molecular docking analysis. In vitro MTT cytotoxic activities on human breast cancer cell line MCF-7 revealed that all the complexes have high cytotoxicity activity (IC50 > 9 µM); furthermore, the selectivity index and SI values were higher (>3). Complex C3 showed the highest cytotoxicity with IC50 = 3.1 µM and SI value (5.55) against MCF7 cell lines and these values were comparable to those of the cisplatin (IC50 and SI values are 5.0 µM and 4.02, respectively). In vivo toxicological assessments on zebrafish embryos revealed that all the Ru-Pt complexes (CI/2/3 ) have poor embryo acute toxic effects over 96 h postfertilization, hpf with LC50 > 65.2 µM. The complex C3 has shown the lowest embryo toxicity (LC50 = 148.8 µM), which is comparable to that of commercial cisplatin (LC50 = 181.1 µM). Based on the cytotoxicity results, complexes C2 and C3 could be considered for further development as chemotherapeutic agents against MCF breast cancer cells.

Trainee life under COVID-19: A systemic case report.

The coronavirus disease 2019 (COVID-19) pandemic has had implications for all of us. For those of us studying on clinical psychology doctorates, and similar psychotherapeutic training courses, this pandemic has led to some particular challenges. This article explores the experiences of a group of clinical psychology trainees, who are also completing intermediate systemic qualification, during the COVID-19 pandemic using a systemic case study approach. We consider the challenges we faced in relation to systemic theory and the intervention we would have used if we were clients in a family therapy clinic. This enabled us to reflect on the importance of self-reflexivity and self-care during these challenging times. Practitioner points: Using systemic thinking and ideas can help organisations make sense of how the system has had to adapt to working during a pandemic and can bring to light some of the challenges.During the pandemic and at other times of crisis, opportunities for students and staff to reflect together are likely to be beneficial.When working independently, as necessitated by the pandemic, finding ways to remain connected to our colleagues will be especially important.Providing opportunities for individuals to give feedback regarding their experiences and to influence organisational change will help individuals gain a sense of agency during a time when they are likely otherwise to feel disempowered.

Therapeutic high affinity T cell receptor targeting a KRASG12D cancer neoantigen.

Neoantigens derived from somatic mutations are specific to cancer cells and are ideal targets for cancer immunotherapy. KRAS is the most frequently mutated oncogene and drives the pathogenesis of several cancers. Here we show the identification and development of an affinity-enhanced T cell receptor (TCR) that recognizes a peptide derived from the most common KRAS mutant, KRASG12D, presented in the context of HLA-A*11:01. The affinity of the engineered TCR is increased by over one million-fold yet fully able to distinguish KRASG12D over KRASWT. While crystal structures reveal few discernible differences in TCR interactions with KRASWT versus KRASG12D, thermodynamic analysis and molecular dynamics simulations reveal that TCR specificity is driven by differences in indirect electrostatic interactions. The affinity enhanced TCR, fused to a humanized anti-CD3 scFv, enables selective killing of cancer cells expressing KRASG12D. Our work thus reveals a molecular mechanism that drives TCR selectivity and describes a soluble bispecific molecule with therapeutic potential against cancers harboring a common shared neoantigen.

2-[4-(Methyl-sulfan-yl)phen-yl]naphtho[1,8-de][1,3,2]diaza-borinane.

The title compound, C(17)H(15)BN(2)S, is one member in a series of diaza-borinanes featuring substitution at the 1-, 2- and 3-positions in the nitro-gen-boron heterocycle. The dihedral angle between the mean planes of the naphthalene and phenyl ring systems is 19.86 (6)°. In the crystal structure, two C-H⋯π inter-actions link the mol-ecules into sheets which lie parallel to the bc plane. There is a π-π inter-action between each pair of centrosymmetrically related sheets [centroid-centroid distance = 3.5922 (8) Å].