Attenuated psychosis symptoms are related to working alliance between therapist and service user.

AIM: Many trials have demonstrated the efficacy of specific therapy modalities for individuals with attenuated psychosis symptoms (APS). Less is known regarding mechanisms behind positive outcomes, including the role of nonspecific therapeutic factors. This study explored working alliance (WA) in a clinic serving individuals with APS to see how WA changed across the course of treatment and its relation to APS. METHODS: Session level APS and WA data was available for 12 individuals of diverse racial and gender identity, (M = 48 sessions each). Multilevel models with random intercepts tested change in WA and APS over time, and cross-sectional and prospective relations. RESULTS: WA increased and APS decreased over time. Cross sectionally, WA and APS were inversely related. Prospective relations were non-significant. CONCLUSION: When symptoms increase, therapists for individuals with APS should be attentive to potential disruptions in WA, though strong WA may be a cross-sectional protective factor.

A Method for Variant Agnostic Detection of SARS-CoV-2, Rapid Monitoring of Circulating Variants, and Early Detection of Emergent Variants Such as Omicron.

The rapid emergence of SARS-CoV-2 variants raised public health questions concerning the capability of diagnostic tests to detect new strains, the efficacy of vaccines, and how to map the geographical distribution of variants to understand transmission patterns and loads on healthcare resources. Next-generation sequencing (NGS) is the primary method for detecting and tracing new variants, but it is expensive, and it can take weeks before sequence data are available in public repositories. This article describes a customizable reverse transcription PCR (RT-PCR)-based genotyping approach which is significantly less expensive, accelerates reporting, and can be implemented in any lab that performs RT-PCR. Specific single-nucleotide polymorphisms (SNPs) and indels were identified which had high positive-percent agreement (PPA) and negative-percent agreement (NPA) compared to NGS for the major genotypes that circulated through September 11, 2021. Using a 48-marker panel, testing on 1,031 retrospective SARS-CoV-2 positive samples yielded a PPA and NPA ranging from 96.3 to 100% and 99.2 to 100%, respectively, for the top 10 most prevalent World Health Organization (WHO) lineages during that time. The effect of reducing the quantity of panel markers was explored, and a 16-marker panel was determined to be nearly as effective as the 48-marker panel at lineage assignment. Responding to the emergence of Omicron, a genotyping panel was developed which distinguishes Delta and Omicron using four highly specific SNPs. The results demonstrate the utility of the condensed panel to rapidly track the growing prevalence of Omicron across the US in December 2021 and January 2022.

The identification of vertical velocity profiles using an inertial sensor to investigate pre-impact detection of falls.

This study investigates distinguishing falls from normal Activities of Daily Living (ADL) by thresholding of the vertical velocity of the trunk. Also presented is the design and evaluation of a wearable inertial sensor, capable of accurately measuring these vertical velocity profiles, thus providing an alternative to optical motion capture systems. Five young healthy subjects performed a number of simulated falls and normal ADL and their trunk vertical velocities were measured by both the optical motion capture system and the inertial sensor. Through vertical velocity thresholding (VVT) of the trunk, obtained from the optical motion capture system, at -1.3 m/s, falls can be distinguished from normal ADL, with 100% accuracy and with an average of 323 ms prior to trunk impact and 140 ms prior to knee impact, in this subject group. The vertical velocity profiles obtained using the inertial sensor, were then compared to those obtained using the optical motion capture system. The signals from the inertial sensor were combined to produce vertical velocity profiles using rotational mathematics and integration. Results show high mean correlation (0.941: Coefficient of Multiple Correlations) and low mean percentage error (6.74%) between the signals generated from the inertial sensor to those from the optical motion capture system. The proposed system enables vertical velocity profiles to be measured from elderly subjects in a home environment where as this has previously been impractical.

A decision-support model for evaluating changes in biopharmaceutical manufacturing processes.

A simulation is described that evaluates the impacts of altering bio-manufacturing processes. Modifications designed to improve production levels, times and costs were assessed, including increasing feed volumes/titres, replacing initial downstream stages with packed or expanded bed affinity steps and removing ion exchange steps. Options were evaluated for manufactured product mass, COG, batch times and development costs and timescales. Metrics were combined using multi-attribute-decision-making techniques generating a single assessment metric for each option. The utility of this approach was illustrated by application to an FDA-approved process manufacturing rattlesnake anti-venom (Protherics U.K.). Currently, ovine serum containing anti-venom IgG is purified by precipitation/centrifugation, prior to antibody proteolysis by papain. An ion exchanger removes F(C), before affinity chromatography yields the final anti-venom. An expanded bed affinity column operating with an 80% higher IgG titre, 66% higher feed volume and without the ion exchanger delivered the best multi-attribute-decision-making value, potentially providing the most desirable alternative.

Pinpointing chiral structures with front-back polarized neutron reflectometry.

A new development in spin-polarized neutron reflectometry enables us to more fully characterize the nucleation and growth of buried domain walls in layered magnetic materials. We applied this technique to a thin-film exchange-spring magnet. After first measuring the reflectivity with the neutrons striking the front, we measure with the neutrons striking the back. Simultaneous fits are sensitive to the presence of spiral spin structures. The technique reveals previously unresolved features of field-dependent domain walls in exchange-spring systems and has sufficient generality to apply to a variety of magnetic systems.

Neuronal signaling through alternative splicing: some exons CaRRE.

Alternative splicing represents a mechanism by which a single gene can be used to create proteins with different functions. Neurons use alternative splicing to produce channels with different sequences and biophysical or regulatory properties. O'Donovan and Darnell discuss a mechanism by which neurons can alter channel splicing in response to neuronal activity through a signal generated by calcium and calcium/calmodulin-dependent kinase activity.

Blockade of NGF-induced neurite outgrowth by a dominant-negative inhibitor of the egr family of transcription regulatory factors.

Although it is well established that members of the Egr family of transcription regulatory factors are induced in many neuronal plasticity paradigms, it is still unclear what role, if any, they play in this process. Because NGF stimulation of pheochromocytoma 12 cells elicits a robust induction of Egr family members, we have investigated their role in mediating long-term effects elicited by NGF in these cells by using the Egr zinc finger DNA-binding domain as a selective antagonist of Egr family-mediated transcription. We report that expression of this Egr inhibitor construct suppresses neurite outgrowth elicited by NGF but not by dibutyryl cAMP. To check that this Egr inhibitor construct does not act by blocking the MEK/ERK pathway, which is known to mediate NGF-induced neurite outgrowth, we confirmed that the Egr inhibitor construct does not block NGF activation of Elk1-mediated transcription, a response that is dependent on this pathway. Conversely, inhibition of MEK does not impair Egr family-mediated transcription. Thus, we conclude (1) that induction of Egr family members and activation of the MEK/ERK pathway by NGF are mediated by separate signaling pathways and (2) that both are required to trigger neurite outgrowth induced by NGF.

Functional comparison of Egr3 transcription factor isoforms: identification of an activation domain in the N-terminal segment absent from Egr3beta, a major isoform expressed in brain.

Recent studies indicate that the Egr family of transcription regulatory factors plays a key role in nervous system development and plasticity. In prior studies, we demonstrated that multiple isoforms of the Egr3 transcription regulatory factor are expressed in brain and appear to be generated by use of alternative translation start sites. To compare the functional activity of these isoforms, we have examined their ability to stimulate transcription of a luciferase reporter construct driven by the Egr response element. Analysis of a series of N-terminal truncation constructs indicates that Egr3 contains two distinct activation domains: one located in the segment upstream of Met(106), the start site of the major truncated isoform Egr3beta, and the other located C-terminal to all of the alternative translation start sites used to generate Egr3 isoforms detected in brain. We confirmed this inference by demonstrating that each of these segments is able to drive transcription when fused to the GAL4 DNA binding domain. Taken together, these studies indicate that the internal translation start sites present in Egr3 are used to generate Egr3 isoforms lacking the activation domain located N-terminal to Met(106).

Legislating for the new predictive genetics.

Many new genetic markers have become available for use in the diagnosis, prognosis or risk prediction of common multifactorial disease such as venous thrombo-embolism, coronary artery disease, dementias and some cancers. Regulation or legislation of their application in the fields of the family, employment, life assurance, confidentiality and property law is required. This is made difficult because of the rapid pace of genetic discoveries and their derived technologies, the diversity of opinions on the legitimate application of these new techniques, and the pluralistic and evolving social norms of society regarding the use of the new genetic methods. This paper examines some of the problems that can arise when regulation is attempted in each of the above fields. A variety of solutions such as referenda, moratoria, ethical codes of professional bodies or the drafting of scientifically accurate and appropriate legislation depending on particular circumstances are discussed as a means of achieving a flexible and responsive approach to the challenges posed by the use of the new genetic markers.

Protection from oxidative stress-induced apoptosis in cortical neuronal cultures by iron chelators is associated with enhanced DNA binding of hypoxia-inducible factor-1 and ATF-1/CREB and increased expression of glycolytic enzymes, p21(waf1/cip1), and erythropoietin.

Iron chelators are pluripotent neuronal antiapoptotic agents that have been shown to enhance metabolic recovery in cerebral ischemia models. The precise mechanism(s) by which these agents exert their effects remains unclear. Recent studies have demonstrated that iron chelators activate a hypoxia signal transduction pathway in non-neuronal cells that culminates in the stabilization of the transcriptional activator hypoxia-inducible factor-1 (HIF-1) and increased expression of gene products that mediate hypoxic adaptation. We examined the hypothesis that iron chelators prevent oxidative stress-induced death in cortical neuronal cultures by inducing expression of HIF-1 and its target genes. We report that the structurally distinct iron chelators deferoxamine mesylate and mimosine prevent apoptosis induced by glutathione depletion and oxidative stress in embryonic cortical neuronal cultures. The protective effects of iron chelators are correlated with their ability to enhance DNA binding of HIF-1 and activating transcription factor 1(ATF-1)/cAMP response element-binding protein (CREB) to the hypoxia response element in cortical cultures and the H19-7 hippocampal neuronal cell line. We show that mRNA, protein, and/or activity levels for genes whose expression is known to be regulated by HIF-1, including glycolytic enzymes, p21(waf1/cip1), and erythropoietin, are increased in cortical neuronal cultures in response to iron chelator treatment. Finally, we demonstrate that cobalt chloride, which also activates HIF-1 and ATF-1/CREB in cortical cultures, also prevents oxidative stress-induced death in these cells. Altogether, these results suggest that iron chelators exert their neuroprotective effects, in part, by activating a signal transduction pathway leading to increased expression of genes known to compensate for hypoxic or oxidative stress.

Major Egr3 isoforms are generated via alternate translation start sites and differ in their abilities to activate transcription.

In previous studies, we detected a major, unidentified Egr response element (ERE) binding complex in brain extracts. We now report that this complex contains a truncated isoform of Egr3 generated by use of an alternate translation start site at methionine 106. Furthermore, the ERE binding complex previously thought to contain full-length Egr3 includes several isoforms generated by initiation at other internal methionines. Full-length and truncated (missing residues 1 to 105) Egr3 isoforms differ in the ability to stimulate transcription directed by a tandem repeat of two EREs but not by a single ERE. Taken together, our results indicate that alternative translation start sites are used to generate Egr3 isoforms with distinct transcriptional properties.

The EGR family of transcription-regulatory factors: progress at the interface of molecular and systems neuroscience.

The EGR family of transcription regulatory factors, which is implicated in orchestrating the changes in gene expression that underlie neuronal plasticity, has attracted the attention of both molecular and systems neuroscientists. In this article, the advances made in both these fields of research are reviewed. Recent systems-based studies underscore the remarkable sensitivity and specificity of the induction of the expression of genes encoding EGR-family members in naturally occurring plasticity paradigms. However, they also challenge conventional views of the role of this family in plasticity. Recent molecular studies have identified the gonadotropin subunit, luteinizing hormone beta, as an EGR1-regulated gene in vivo and uncovered an essential role for EGR3 in muscle-spindle development. In addition, the discovery of novel proteins that are capable of suppressing EGR-mediated transcription cast doubt over the prevalent assumption that changes in EGR mRNA or protein levels provide an accurate measure of EGR-driven transcriptional activity.

Sequential expression of Egr-1 and Egr-3 in hippocampal granule cells following electroconvulsive stimulation.

Previous studies examining the regulation of immediate early gene mRNAs by neuronal stimulation have revealed that two members of the Egr family of transcription factors, Egr-1 and Egr-3, display parallel response patterns. As these transcription factors compete for the same consensus sequence, we investigated how their expression and DNA binding activities are coordinated. Following electroconvulsive stimulation, which induces rapid increases in both Egr-1 and Egr-3 mRNA levels in dentate granule cells, we found that these proteins are induced sequentially. Egr-1 protein levels peak at 0.5-1 h and decay to basal levels by 4 h. In contrast, Egr-3 protein levels respond more slowly; little change is apparent at 1 h, and peak levels are not reached until 4 h following stimulation. Gel shift assays demonstrated that Egr-1 and Egr-3 DNA binding activities follow the same pattern. These findings indicate that Egr-1 and Egr-3 act in concert to mediate early and late phases, respectively, of the transcriptional response regulated by their cognate response element.

Removal of an N-terminal peptide from mitochondrial aspartate aminotransferase abolishes its interactions with mitochondria in vitro.

Treatment of mitochondrial aspartate aminotransferase from rat liver with trypsin leads to specific cleavage of the bonds between residues 26 and 27, and residues 31 and 32. The proteolysed enzyme has only a small residual catalytic activity, but retains a conformation similar to that of the native form as judged by accessibility and reactivity of cysteine residues. Proteolysis abolishes the ability of the enzyme either to bind to mitochondria or to be imported into the organelles. This suggests that the N-terminal segment of the native enzyme is essential for both of these functions, at least in the model system used to study the import process.