In situ phytoextraction of Mn and NH4+-N from aqueous electrolytic manganese residue solution by Pistia stratiotes: Effects of Fe/Co presence and rhizospheric microbe synergistic involvement.

The electrolytic manganese industry produces a large amount of electrolytic manganese residue (EMR). Soluble Mn, NH4+-N, and other pollutants may be released from the open-air stacked EMR and transported to the environment along with rainfall or surface runoff. Aqueous EMR solution (AES) generally contains various elements required for plant growth, and phytoremediation can be applied to remove these pollutants from AES. Since the contents of Fe and Co vary greatly in AES depending on the ore sources as well as the pre-treatment processes, the presence of bioavailable Fe and Co at different levels may affect plant growth, the rhizosphere microbes, and pollutant removal. The present study investigated the in-situ removal of Mn(II) and NH4+-N from AES solution using free floating aquatic plant Pistia stratiotes, focusing especially on the effects of Fe/Co presence and rhizospheric microbe synergistic involvement on contaminant removal. The results showed that 69.08% of Mn and 94.99% of NH4+-N were removed by P. stratiotes in 24 d. Both the presence of Fe(II) and Co(II) facilitated the Mn(II) immobilization and increased Mn(II) removal by 19-31% due to the enhanced peroxidase activity and the increased Mn accumulating in roots The complete removal of Mn from AES was found in the presence of Fe(II) at 2 mg L-1 or Co(II) at 0.5 mg L-1 and more than 51% accumulated Mn in the roots was stored in the vacuole and cytoplasm. BioMnOx was found on the surface of the roots, revealing that rhizofiltration, rhizospheric plaque/biofilm formation, and Mn biogeochemical cycle exert synergic effects on Mn(II) immobilization. The findings of the present study demonstrate the feasibility of using P. stratiotes in the treatment of aqueous EMR solutions and the presence of an appropriate amount of bio-available Fe and Co can promote the removal of Mn(II) and NH4+-N.

Diesel degradation capability and environmental robustness of strain Pseudomonas aeruginosa WS02.

Petroleum hydrocarbon (PHC) degrading bacteria have been frequently discovered. However, in practical application, a single species of PHC degrading bacterium with weak competitiveness may face environmental pressure and competitive exclusion due to the interspecific competition between petroleum-degrading bacteria as well as indigenous microbiota in soil, leading to a reduced efficacy or even malfunction. In this study, the diesel degradation ability and environmental robustness of an endophytic strain Pseudomonas aeruginosa WS02, were investigated. The results show that the cell membrane surface of WS02 was highly hydrophobic, and the strain secreted glycolipid surfactants. Genetic analysis results revealed that WS02 contained multiple metabolic systems and PHC degradation-related genes, indicating that this strain theoretically possesses the capability of oxidizing both alkanes and aromatic hydrocarbons. Gene annotation also showed many targets which coded for heavy metal resistant and metal transporter proteins. The gene annotation-based inference was confirmed by the experimental results: GC-MS analysis revealed that short chain PHCs (C10-C14) were completely degraded, and the degradation of PHCs ranging from C15-C22 were above 90% after 14 d in diesel-exposed culture; Heavy metal (Mn2+, Pb2+ and Zn2+) exposure was found to affect the growth of WS02 to some extent, but not its ability to degrade diesel, and the degradation efficiency was still maintained at 39-59%. WS02 also showed a environmental robustness along with PHC-degradation performance in the co-culture system with other bacterial strains as well as in the co-cultured system with the indigenous microbiota in soil fluid extracted from a PHC-contaminated site. It can be concluded that the broad-spectrum diesel degradation efficacy and great environmental robustness give P. aeruginosa WS02 great potential for application in the remediation of PHC-contaminated soil.

What treatment outcomes matter most? A Q-study of outcome priority profiles among youth with lived experience of depression.

Interest in youth perspectives on what constitutes an important outcome in the treatment of depression has been growing, but limited attention has been given to heterogeneity in outcome priorities, and minority viewpoints. These are important to consider for person-centred outcome tracking in clinical practice, or when conducting clinical trials targeting specific populations. This study used Q-methodology to identify outcome priority profiles among youth with lived experience of service use for depression. A purposive sample of 28 youth (aged 16-21 years) rank-ordered 35 outcome statements by importance and completed brief semi-structured interviews eliciting their sorting rationales. By-person principal component analysis was used to identify outcome priority profiles based on all Q-sort configurations. Priority profiles were described and interpreted with reference to the qualitative interview data. Four distinct outcome priority profiles were identified: "Relieving distress and experiencing a happier emotional state"; "Learning to cope with cyclical distressing emotional states"; "Understanding and processing distressing emotional states"; and "Reduced interference of ongoing distressing emotional states with daily life". All four profiles prioritised improvements in mood and the ability to feel pleasure but differed in the level of importance assigned to learning coping skills, processing experiences, and the reduced interference of depression with life and identity. As part of a person-centered approach to care delivery, care providers should routinely engage young people in conversation and shared decision-making about the types of change they would like to prioritise and track during treatment, beyond a common core of consensus outcomes.

Determination of Markers of Successful Implementation of Mental Health Apps for Young People: Systematic Review.

BACKGROUND: Smartphone apps have the potential to address some of the current issues facing service provision for young people's mental health by improving the scalability of evidence-based mental health interventions. However, very few apps have been successfully implemented, and consensus on implementation measurement is lacking. OBJECTIVE: This review aims to determine the proportion of evidence-based mental health and well-being apps that have been successfully adopted and sustained in real-world settings. A secondary aim is to establish if key implementation determinants such as coproduction, acceptability, feasibility, appropriateness, and engagement contribute toward successful implementation and longevity. METHODS: Following the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines, an electronic search of 5 databases in 2021 yielded 18,660 results. After full-text screening, 34 articles met the full eligibility criteria, providing data on 29 smartphone apps studied with individuals aged 15 to 25 years. RESULTS: Of 34 studies, only 10 (29%) studies were identified that were evaluating the effectiveness of 8 existing, commercially available mental health apps, and the remaining 24 (71%) studies reported the development and evaluation of 21 newly developed apps, of which 43% (9/21) were available, commercially or otherwise (eg, in mental health services), at the time of enquiry. Most studies addressed some implementation components including adoption, acceptability, appropriateness, feasibility, and engagement. Factors including high cost, funding constraints, and lengthy research processes impeded implementation. CONCLUSIONS: Without addressing common implementation drivers, there is considerable redundancy in the translation of mobile mental health research findings into practice. Studies should embed implementation strategies from the outset of the planned research, build collaborations with partners already working in the field (academic and commercial) to capitalize on existing interventions and platforms, and modify and evaluate them for local contexts or target problems and populations. TRIAL REGISTRATION: PROSPERO CRD42021224365; https://tinyurl.com/4umpn85f.

A Transdiagnostic, Emotion Regulation App (Eda) for Children: Design, Development, and Lessons Learned.

BACKGROUND: Digital interventions, including mobile apps, represent a promising means of providing effective mental health support to children and young people. Despite the increased availability of mental health apps, there is a significant gap for this age group, especially for children (aged 10-12 years). Research investigating the effectiveness and development process of child mental health apps is limited, and the field faces persistent issues in relation to low user uptake and engagement, which is assumed to be a result of limited user involvement in the design process. OBJECTIVE: This study aims to present the development and design process of a new mental health app for children that targets their emotion regulation abilities. We describe the creation of a new interdisciplinary development framework to guide the design process and explain how each activity informed different app features. METHODS: The first 2 stages of the framework used a variety of methods, including weekly classroom observations over a 6-month period (20 in total); public engagement events with the target group (N=21); synthesis of the existing evidence as part of a meta-analysis; a series of co-design and participatory workshops with young users (N=33), clinicians (N=7), researchers (N=12), app developers (N=1), and designers (N=2); and finally, testing of the first high-tech prototype (N=15). RESULTS: For the interdisciplinary framework, we drew on methods derived from the Medical Research Council framework for complex interventions, the patient-clinician framework, and the Druin cooperative inquiry. The classroom observations, public engagement events, and synthesis of the existing evidence informed the first key pillars of the app and wireframes. Subsequently, a series of workshops shaped and reshaped the content and app features, including games, psychoeducational films, and practice modules. On the basis of the prototype testing sessions, we made further adjustments to improve the app. CONCLUSIONS: Although mobile apps could be highly suitable to support children's mental health on a wider scale, there is little guidance on how these interventions could be designed and developed. The involvement of young users across different design activities is very valuable. We hope that our interdisciplinary framework and description of the used methods will be helpful to others who are hoping to develop mental health apps for children and young people.

Applying behaviour change theory to understand the barriers to implementing routine outcome monitoring.

BACKGROUND: Routine outcome monitoring (ROM) is a valuable tool for monitoring client progress and pre-empting deterioration, however, there is considerable variation in how data are collected and recorded and uptake in clinical practice remains low. The aim of this study was to develop a self-report measure of practitioner attitudes to ROM in order to better understand the barriers to successful implementation in Child and Adolescent Mental Health Services (CAMHS). METHODS: An anonymous survey was completed by 184 CAMHS practitioners in the United Kingdom. The survey was designed using the Capability, Opportunity, and Motivation Model of Behaviour (COM-B). Practitioners who reported using ROM frequently in their clinical work (53%) were compared to those who used ROM infrequently (47%) across dimensions of the COM-B survey subscales. RESULTS: Confirmatory factor analysis confirmed the proposed four-factor structure, showing acceptable model fit, with high factor loadings and good reliability for all subscales. Frequent users of ROM exhibited significantly higher psychological capability, physical opportunity, social opportunity, and motivation, compared to infrequent users F (4, 140) = 14.76, p < .0001; Pillai's Trace = .297, partial η2 = .30. Results highlight several barriers to ROM, including the belief that there is not a strong evidence base for ROM, not receiving external training, and not discussing feedback and outcome data in supervision. IMPLICATIONS: In the hope of improving the successful implementation of ROM, this research provides an evidence-based tool for assessing practitioners' attitudes to ROM, which map on to intervention functions and represent targets for future implementation efforts. PRACTITIONER POINTS: The value of routine outcome monitoring (ROM) as a means to measure client progress and to elevate the efficiency and quality of mental health care is well-documented in the research literature, however, uptake in practice remains relatively low. This study applied behaviour change theory to develop a psychometrically sound self-report measure of practitioners' perspectives and practices to understand the barriers to implementation in child and adolescent mental health services in the United Kingdom. The complex and multifaceted nature of the barriers to implementation requires multilevel behaviour change strategies at the client, clinician, and organisational level. Recommendations for practice include the need for integrated, multilevel strategies aimed at improving practitioners' capabilities and motivations, strong organisational leadership and a culture of data gathering and sharing, and implementation interventions, which are tailored to target local barriers.

How to manage endings in unsuccessful therapy: A qualitative comparison of youth and clinician perspectives.

Background: Approximately half of those who access child and adolescent mental health services do not show measurable improvement in symptoms. This study aimed to provide practice recommendations for managing treatment endings, particularly when outcomes have not improved. Method: Semi-structured interviews were carried out with 26 young people with a history of anxiety and/or depression along with 7 roundtable sessions with 52 mental health clinicians. Data were analyzed using Framework Analysis. Results: A common experience for young people when outcomes did not improve was a poor experience of the treatment ending, which often resulted in setbacks in their mental health and feelings of loss and abandonment. Clinicians agreed that ending was hard for young people and reported that they found managing ending hard on a personal and professional level. This was compounded by unrealistically high public expectations about the impact of therapy on outcomes and trying to strike a balance between fostering hope and managing expectations, within a context of inflexible service structures and resource constraint. Implications: Recommendations include establishing expectations from the outset and a shared understanding of what outcomes matter most to the young person. This can be achieved through communicating honestly about likely outcomes, while also providing hope.

Understanding the illness representations of young people with anxiety and depression: A qualitative study.

BACKGROUND: Many young people with anxiety or depression drop out of treatment early, and/or leave treatment without showing measurably improved symptom levels. To enhance treatment engagement and effectiveness, it is critical to better understand how young people's perceptions of the symptoms, causes, consequences, treatability, and course of their anxiety and depression influence engagement. AIM: This study aimed to provide a qualitative account of illness perceptions among youth with anxiety and depression by applying the Common Sense Model of Self-Regulation (CSM), which was developed in physical health contexts. METHODS: Semi-structured interviews were conducted with 26 young people (aged 16-24, 73% female) with a history of anxiety and/or depression. Interviews were analysed using a combination of theory- and data-driven analysis techniques, consisting primarily of deductive thematic analysis. RESULTS: The five themes broadly mapped onto the dimensions of the CSM, suggesting parallels in how mental and physical health problems are perceived. Anxiety and depression were viewed as non-linear, relapsing and remitting, but lifelong conditions, with a fluctuating and complex path to recovery and coping. Youth described pervasive negative impacts on their lives, but also described some positive aspects. IMPLICATIONS: Better understanding of young people's illness beliefs has the potential to open a range of intervention possibilities by prioritizing young people's illness perceptions over the clinician's understanding and the supposed objective condition severity and trajectory. Although this study supported a common structure of illness beliefs, the content of these beliefs was idiosyncratic and specific to anxiety and depression, suggesting the need to develop a valid tool to measure illness perceptions in this group. PRACTITIONER POINTS: Our findings suggest that illness perceptions are complex, highly idiosyncratic, and specific to youth anxiety and depression. Given the complexity of these beliefs and the known association with important treatment- and health-related outcomes, it is important that clinical formulation incorporates young people's illness belief models, including their perceptions of symptoms, cause, timeline to recovery, consequences, and personal and treatment control. To increase help-seeking, treatment engagement and adaptive coping strategies, therapy should work to a shared understanding of illness beliefs. Increasing congruence between the belief models of young people, families, and clinicians may serve to improve treatment benefits and address the unmet mental health needs of young people.

Systematic Review and Meta-analysis: Outcomes of Routine Specialist Mental Health Care for Young People With Depression and/or Anxiety.

OBJECTIVE: Depression and anxiety are the most prevalent mental health problems in youth, yet almost nothing is known about what outcomes are to be expected at the individual level following routine treatment. This paper sets out to address this gap by undertaking a systematic review of outcomes following treatment as usual (TAU) with a particular focus on individual-level outcomes. METHOD: MEDLINE, Embase and PsycInfo were searched for articles published between 1980 and January 2019 that assessed TAU outcomes for youth depression and anxiety accessing specialist mental health care. Meta-analysis considered change at both group-level pre-post effect size (ES) and individual-level recovery, reliable change, and reliable recovery. Temporal analysis considered stability of primary and secondary outcomes over time. Subgroup analysis considered the moderating effect of informant; presenting problem; study design; study year; mean age of youth; use of medication; intervention dosage and type of treatment offered on outcomes. A protocol was preregistered on PROSPERO (CRD42017063914). RESULTS: Initial screening of 6,350 publications resulted in 38 that met the inclusion criteria, and that were subsequently included in meta-analyses. This resulted in a final full pooled sample of 11,739 young people (61% of whom were female, mean age 13.8 years). The pre-post ES (Hedges' g) at first/final outcome (13/26 weeks) was -0.74/-0.87. The individual-level change on measures of self-report was 38% reliable improvement, 44% no reliable change, and 6% reliable deterioration. Outcomes varied according to moderators, informant, problem type and dosage. CONCLUSION: Poor data quantity and quality are limitations, but this is the first study that indicates likely rates of reliable improvement for those accessing TAU. We propose the need for improved reporting of both individual-level metrics and details of TAU to enable greater understanding of likely current outcomes from routine care for youths with depression and anxiety in order to allow the potential for further improvement of impact.

Evolution of Excitation-Contraction Coupling.

In mammalian cardiomyocytes, Ca2+ influx through L-type voltage-gated Ca2+ channels (VGCCs) is amplified by release of Ca2+ via type 2 ryanodine receptors (RyR2) in the sarcoplasmic reticulum (SR): a process termed Ca2+-induced Ca2+-release (CICR). In mammalian skeletal muscles, VGCCs play a distinct role as voltage-sensors, physically interacting with RyR1 channels to initiate Ca2+ release in a mechanism termed depolarisation-induced Ca2+-release (DICR). In the current study, we surveyed the genomes of animals and their close relatives, to explore the evolutionary history of genes encoding three proteins pivotal for ECC: L-type VGCCs; RyRs; and a protein family that anchors intracellular organelles to plasma membranes, namely junctophilins (JPHs). In agreement with earlier studies, we find that non-vertebrate eukaryotes either lack VGCCs, RyRs and JPHs; or contain a single homologue of each protein. Furthermore, the molecular features of these proteins thought to be essential for DICR are only detectable within vertebrates and not in any other taxonomic group. Consistent with earlier physiological and ultrastructural observations, this suggests that CICR is the most basal form of ECC and that DICR is a vertebrate innovation. This development was accompanied by the appearance of multiple homologues of RyRs, VGCCs and junctophilins in vertebrates, thought to have arisen by 'whole genome replication' mechanisms. Subsequent gene duplications and losses have resulted in distinct assemblies of ECC components in different vertebrate clades, with striking examples being the apparent absence of RyR2 from amphibians, and additional duplication events for all three ECC proteins in teleost fish. This is consistent with teleosts possessing the most derived mode of DICR, with their Cav1.1 VGCCs completely lacking in Ca2+ channel activity.

Review: What Outcomes Count? A Review of Outcomes Measured for Adolescent Depression Between 2007 and 2017.

OBJECTIVE: Measurement of treatment outcomes in childhood depression has traditionally focused on assessing symptoms from the clinician's perspective, without exploring other outcome domains or considering young people's perspectives. This systematic review explored the extent to which multidimensional and multi-informant outcome measurements have been used in clinical research for adolescent depression in the past decade and how patterns have evolved over time. METHOD: Embase, Medline, and PsycINFO were searched, and studies that were published from 2007 through 2017 and assessed the effectiveness of treatments or service provision for adolescent depression were included. Of 7,483 studies screened, 95 met the inclusion criteria and were included for data extraction and analysis. RESULTS: Ten outcomes domains were identified, 2 of which were assessed on average using 4 outcome measures. Most studies (94%) measured symptoms, followed by functioning (52%). Other domains such as personal growth, relationships, quality of life, and service satisfaction were each considered by less than 1 in 10 studies. Youth self-report was considered by 54% but tended to focus on secondary outcomes. Multidimensional and multi-informant measurements were more frequent in studies published since 2015. CONCLUSION: Recent clinical research continues to prioritize symptoms measurement based on clinician report and has not yet fully embraced multidimensional and multi-informant approaches. In the context of significant policy shifts toward patient-centered and evidence-based care, measuring what matters most to patients has become a priority, but this is not yet widely reflected in clinical research.

Enzalutamide inhibits testosterone-induced growth of human prostate cancer xenografts in zebrafish and can induce bradycardia.

The zebrafish has become a popular human tumour xenograft model, particularly for solid tumours including prostate cancer (PCa). To date PCa xenotransplantation studies in zebrafish have not been performed in the presence of testosterone, even when employing androgen-dependent cell models, such as the LNCaP cell line. Thus, with the goal of more faithfully modelling the hormonal milieu in which PCa develops in humans, we sought to determine the effects of exogenous testosterone on the growth of LNCaP, or androgen-independent C4-2 cells xenografted into zebrafish embryos. Testosterone significantly increased engrafted LNCaP proliferation compared to control xenografts, which could be inhibited by co-administration of the anti-androgen receptor drug, enzalutamide. By contrast, C4-2 cell growth was not affected by either testosterone or enzalutamide. Enzalutamide also induced bradycardia and death in zebrafish embryos in a dose-dependent manner and strongly synergized with the potassium-channel blocking agent, terfenadine, known to induce long QT syndrome and cardiac arrhythmia. Together, these data not only indicate that testosterone administration should be considered in all PCa xenograft studies in zebrafish but also highlights the unique opportunity of this preclinical platform to simultaneously evaluate efficacy and toxicity of novel therapies and/or protective agents towards developing safer and more effective PCa treatments.

Estrogen receptor alpha (ESR1)-signaling regulates the expression of the taxane-response biomarker PRP4K.

The pre-mRNA splicing factor 4 kinase PRP4K (PRPF4B), is an essential kinase that is a component of the U5 snRNP and functions in spliceosome assembly. We demonstrated that PRP4K is a novel biological marker for taxane response in ovarian cancer patients and reduced levels of PRP4K correlate with intrinsic and acquired taxane resistance in both breast and ovarian cancer. Breast cancer treatments are chosen based on hormone and growth factor receptor status, with HER2 (ERBB2) positive breast cancer patients receiving anti-HER2 agents and taxanes and estrogen receptor alpha (ESR1) positive (ER+) breast cancer patients receiving anti-estrogen therapies such as tamoxifen. Here we demonstrate that PRP4K is expressed in the normal mammary duct epithelial cells of the mouse, and that estrogen induces PRP4K gene and protein expression in ER+ human MCF7 breast cancer cells. Estrogen acts through ESR1 to regulate PRP4K expression, as over-expression of ESR1 in the ER-negative MDA-MB-231 breast cancer cell line increased the expression of this kinase, and knock-down of ESR1 in ER+ T47D breast cancer cells reduced PRP4K levels. Furthermore, treatment with 4-hydroxytamoxifen (4-OHT) resulted in a dose-dependent decrease in PRP4K protein expression in MCF7 cells. Consistent with our previous studies identifying PRP4K as a taxane-response biomarker, reduced PRP4K expression in 4-OHT-treated cells correlated with reduced sensitivity to paclitaxel. Thus, PRP4K is novel estrogen regulated kinase, and its levels can be reduced by 4-OHT in ER+ breast cancer cells altering their response to taxanes.

Connecting the speckles: Splicing kinases and their role in tumorigenesis and treatment response.

Alternative pre-mRNA splicing in higher eukaryotes enhances transcriptome complexity and proteome diversity. Its regulation is mediated by a complex RNA-protein network that is essential for the maintenance of cellular and tissue homeostasis. Disruptions to this regulatory network underlie a host of human diseases and contribute to cancer development and progression. The splicing kinases are an important family of pre-mRNA splicing regulators, , which includes the CDC-like kinases (CLKs), the SRSF protein kinases (SRPKs) and pre-mRNA splicing 4 kinase (PRP4K/PRPF4B). These splicing kinases regulate pre-mRNA splicing via phosphorylation of spliceosomal components and serine-arginine (SR) proteins, affecting both their nuclear localization within nuclear speckle domains as well as their nucleo-cytoplasmic shuttling. Here we summarize the emerging evidence that splicing kinases are dysregulated in cancer and play important roles in both tumorigenesis as well as therapeutic response to radiation and chemotherapy.

Comparison of senescence-associated miRNAs in primary skin and lung fibroblasts.

MicroRNAs are non-coding RNAs with roles in many cellular processes. Tissue-specific miRNA profiles associated with senescence have been described for several cell and tissue types. We aimed to characterise miRNAs involved in core, rather than tissue-specific, senescence pathways by assessment of common miRNA expression differences in two different cell types, with follow-up of predicted targets in human peripheral blood. MicroRNAs were profiled in early and late passage primary lung and skin fibroblasts to identify commonly-deregulated miRNAs. Expression changes of their bioinformatically-predicted mRNA targets were then assessed in both cell types and in human peripheral blood from elderly participants in the InCHIANTI study. 57/178 and 26/492 microRNAs were altered in late passage skin and lung cells respectively. Three miRNAs (miR-92a, miR-15b and miR-125a-3p) were altered in both tissues. 14 mRNA targets of the common miRNAs were expressed in lung and skin fibroblasts, of which two demonstrated up-regulation in late passage skin and lung cells (LYST; p = 0.02 [skin] and 0.02 [lung] INMT; p = 0.03 [skin] and 0.04 [lung]). ZMPSTE24 and LHFPL2 demonstrated altered expression in late passage skin cells only (p = 0.01 and 0.05 respectively). LHFPL2 was also positively correlated with age in peripheral blood (p value = 6.6 × 10(-5)). We find that the majority of senescence-associated miRNAs demonstrate tissue-specific effects. However, miRNAs showing common effects across tissue types may represent those associated with core, rather than tissue-specific senescence processes.

Splicing factor 3B1 hypomethylation is associated with altered SF3B1 transcript expression in older humans.

Ageing in man is associated with changes to the splicing factor pool. A proportion of splicing factors are regulated during ageing by mechanisms involving the Ataxia Telangiectasia Mutated (ATM) gene, but the factors that determine the remaining proportion have yet to be identified. DNA methylation is known to be an important regulatory mechanism of gene expression. We assessed age-associated methylation and expression levels for 27 splicing factor genes, in peripheral blood samples from the InCHIANTI study. Examination of splicing patterns at specific loci was examined in a second cohort, the Exeter 10000 study. 27/502 methylation probes in 17 different genes were associated with age. Most changes were not associated with transcript expression levels or splicing patterns, but hypomethylation of the SF3B1 promoter region was found to mediate 53% of the relationship between age and transcript expression at this locus (p=0.02). DNA methylation does not appear to play a major role in regulation of the splicing factors, but changes in SF3B1 expression may be attributable to promoter hypomethylation at this locus. SF3B1 encodes a critical component of the U2 snRNP; altered expression of this gene may therefore contribute to the loss of regulated mRNA splicing that occurs with age.

Changes in splicing factor expression are associated with advancing age in man.

Human ageing is associated with decreased cellular plasticity and adaptability. Changes in alternative splicing with advancing age have been reported in man, which may arise from age-related alterations in splicing factor expression. We determined whether the mRNA expression of key splicing factors differed with age, by microarray analysis in blood from two human populations and by qRT-PCR in senescent primary fibroblasts and endothelial cells. Potential regulators of splicing factor expression were investigated by siRNA analysis. Approximately one third of splicing factors demonstrated age-related transcript expression changes in two human populations. Ataxia Telangiectasia Mutated (ATM) transcript expression correlated with splicing factor expression in human microarray data. Senescent primary fibroblasts and endothelial cells also demonstrated alterations in splicing factor expression, and changes in alternative splicing. Targeted knockdown of the ATM gene in primary fibroblasts resulted in up-regulation of some age-responsive splicing factor transcripts. We conclude that isoform ratios and splicing factor expression alters with age in vivo and in vitro, and that ATM may have an inhibitory role on the expression of some splicing factors. These findings suggest for the first time that ATM, a core element in the DNA damage response, is a key regulator of the splicing machinery in man.

Towards a gene expression biomarker set for human biological age.

We have previously described a statistical model capable of distinguishing young (age <65 years) from old (age ≥75 years) individuals. Here we studied the performance of a modified model in three populations and determined whether individuals predicted to be biologically younger than their chronological age had biochemical and functional measures consistent with a younger biological age. Those with 'younger' gene expression patterns demonstrated higher muscle strength and serum albumin, and lower interleukin-6 and blood urea concentrations relative to 'biologically older' individuals (odds ratios 2.09, 1.64, 0.74, 0.74; P = 2.4 × 10(-2) , 3.5 × 10(-4) , 1.8 × 10(-2) , 1.5 × 10(-2) , respectively). We conclude that our expression signature of age is robust across three populations and may have utility for estimation of biological age.

Human aging is characterized by focused changes in gene expression and deregulation of alternative splicing.

Aging is a major risk factor for chronic disease in the human population, but there are little human data on gene expression alterations that accompany the process. We examined human peripheral blood leukocyte in-vivo RNA in a large-scale transcriptomic microarray study (subjects aged 30-104 years). We tested associations between probe expression intensity and advancing age (adjusting for confounding factors), initially in a discovery set (n= 58), following-up findings in a replication set (n=240). We confirmed expression of key results by real-time PCR. Of 16,571 expressed probes, only 295 (2%) were robustly associated with age. Just six probes were required for a highly efficient model for distinguishing between young and old (area under the curve in replication set; 95%). The focused nature of age-related gene expression may therefore provide potential biomarkers of aging. Similarly, only 7 of 1065 biological or metabolic pathways were age-associated, in gene set enrichment analysis, notably including the processing of messenger RNAs (mRNAs); [P<0.002, false discovery rate (FDR) q<0.05]. This is supported by our observation of age-associated disruption to the balance of alternatively expressed isoforms for selected genes, suggesting that modification of mRNA processing may be a feature of human aging.

Nanofiber assembly by rotary jet-spinning.

High-voltage electrical fields and low production rate limit electrospinning, the electrical charging of polymer liquids, as a means of nanofiber fabrication. Here, we show a facile method of fabrication of aligned three-dimensional nanofiber structures by utilizing high-speed, rotating polymer solution jets to extrude fibers. Termed rotary jet-spinning, fiber morphology, diameter, and web porosity can be controlled by varying nozzle geometry, rotation speed, and polymer solution properties. We demonstrate the utility of this technique for tissue engineering by building anisotropic arrays of biodegradable polymer fibers and seeding the constructs with neonatal rat ventricular cardiomyocytes. The myocytes used the aligned fibers to orient their contractile cytoskeleton and to self-organize into a beating, multicellular tissue that mimics the laminar, anisotropic architecture of the heart muscle. This technique may prove advantageous for building uniaxially aligned nanofiber structures for polymers which are not amenable to fabrication by electrospinning.