Actionable Steps for Preventing Firearm Suicides among Adolescents and Young Adults.

The suicide rate among adolescents and young adults is on the rise once again, with firearm suicides accounting for more than two-thirds of suicides among North Carolinians aged 15-24. Reversing this trend will require a multifaceted strategy involving providers, gun owners, and sound policy decisions.

In-depth characterization of Trichoderma reesei cellobiohydrolase TrCel7A produced in Nicotiana benthamiana reveals limitations of cellulase production in plants by host-specific post-translational modifications.

Sustainable production of biofuels from lignocellulose feedstocks depends on cheap enzymes for degradation of such biomass. Plants offer a safe and cost-effective production platform for biopharmaceuticals, vaccines and industrial enzymes boosting biomass conversion to biofuels. Production of intact and functional protein is a prerequisite for large-scale protein production, and extensive host-specific post-translational modifications (PTMs) often affect the catalytic properties and stability of recombinant enzymes. Here we investigated the impact of plant PTMs on enzyme performance and stability of the major cellobiohydrolase TrCel7A from Trichoderma reesei, an industrially relevant enzyme. TrCel7A was produced in Nicotiana benthamiana using a vacuum-based transient expression technology, and this recombinant enzyme (TrCel7Arec ) was compared with the native fungal enzyme (TrCel7Anat ) in terms of PTMs and catalytic activity on commercial and industrial substrates. We show that the N-terminal glutamate of TrCel7Arec was correctly processed by N. benthamiana to a pyroglutamate, critical for protein structure, while the linker region of TrCel7Arec was vulnerable to proteolytic digestion during protein production due to the absence of O-mannosylation in the plant host as compared with the native protein. In general, the purified full-length TrCel7Arec had 25% lower catalytic activity than TrCel7Anat and impaired substrate-binding properties, which can be attributed to larger N-glycans and lack of O-glycans in TrCel7Arec . All in all, our study reveals that the glycosylation machinery of N. benthamiana needs tailoring to optimize the production of efficient cellulases.

Responding to Adolescents at Risk of Suicide: Implications of the Ideation-to-Action Framework.

Though suicide is the second leading cause of death among adolescents, it continues to be difficult to predict. Recent research has led to the development of the ideation-to-action framework, which provides a new conceptualization of suicide risk. The implications of this approach for assessment and intervention in medical settings are discussed.

Mountaintop Removal Coal Mining and Emergent Cases of Psychological Disorder in Kentucky.

Although mountaintop removal (MTR) coal extraction techniques have been employed in Appalachia for decades, relatively little research has examined its potential psychological impact on people living in close proximity to MTR activity. The current study taps the State Emergency Department Database (Healthcare Cost and Utilization Project, Kentucky State Emergency Department Database, 2008) to examine the relative risk for diagnoses of depressive, substance use, and anxiety disorders originating in areas with and without MTR activity. Logistical regression analyses, controlling for ethnicity, rurality, mean income, and gender, indicated that MTR independently predicts greater risk for depressive (OR 1.37) and substance use disorders (OR 1.41), but not anxiety disorders. Overall, these findings have public health policy implications, build on other evidence of increased risk of negative mental health outcomes related to MTR, and lend some support to the validity of solastalgia related to environmental change.

Circulating plasma microRNAs as a screening method for detection of colorectal adenomas.

BACKGROUND: MicroRNAs (miRNAs) are small non-coding RNA molecules. Reduced or increased levels of specific miRNAs are observed in colon and other cancers, supporting their role in carcinogenesis. Detection of colorectal polyps is the cornerstone of the Bowel Cancer Screening Programme in the UK. However, uptake of screening nationally remains under 60%. We aimed to see whether circulating plasma miRNAs can be used to screen for patients with colorectal polyps, adenomas, or both. METHODS: Blood samples were taken from patients from the Bowel Cancer Screening Programme (asymptomatic but faecal occult blood testing [FOBt] positive). Plasma RNA was extracted, target miRNAs (19a, 98, 146b, 186, 191, 222*, 331-5p, 452, 625, 664, 1247) were identified on pooled case miRNA assay cards, and miRNA fraction was quantified by quantitative RT-PCR assay. Results were compared with endoscopy reports and with histology of any polyps identified and removed. Analysis was done with Excel (2011) and SPSS (version 20) software. FINDINGS: 210 patients were included (117 with polyps, 12 with cancer, 81 healthy controls [FOBt positive]). The miRNA panel showed significant differences in expression (on t testing) for patients compared with controls for those with polyps, cancer, or both (miR-19a, p=0·0184; miR-98, p=0·0206; miR-146b, p=0·0029; miR-186, p=0·0006; miR-62,5 p=0·0008), polyps (miR-19a, p=0·0233; miR-98, p=0·0224; miR-146b, p=0·003; miR-186, p=0·0004; miR-625, p=0·001), adenomas (miR-19a, p=0·0339; miR-98, p=0·0266; miR-146b, p=0·0045; miR-186, p=0·0008; miR-625, p=0·0049), multiple adenomas (both sides of colon; miR-146b, p=0·0194; miR-186, p=0·0226; miR-625, p=0·0013), and right-sided adenomas (miR-98, p=0·031; miR-146b, p=0·0076; miR-186, p=0·0041; miR-331-5p, p=0·0142; miR-625, p=0·0049). Receiver operating characteristic analysis showed sensitivity of 60% or more, and specificity of 86% or more for men with polyps, men with adenomas, all patients with haemorrhoids or diverticulosis and polyps, and all patients with haemorrhoids or diverticulosis and adenomas. INTERPRETATION: The target miRNAs that we identified showed significant differences in expression levels for patients with polyps and patients with adenomas from controls. Use of this panel has potential as a screening test. FUNDING: Bowel Disease Research Foundation.

Design and validation of bending test method for characterization of miniature pediatric cortical bone specimens.

Osteogenesis imperfecta is a genetic disorder of bone fragility; however, the effects of this disorder on bone material properties are not well understood. No study has yet measured bone material strength in humans with osteogenesis imperfecta. Small bone specimens are often extracted during routine fracture surgeries in children with osteogenesis imperfecta. These specimens could provide valuable insight into the effects of osteogenesis imperfecta on bone material strength; however, their small size poses a challenge to their mechanical characterization. In this study, a validated miniature three-point bending test is described that enables measurement of the flexural material properties of pediatric cortical osteotomy specimens as small as 5 mm in length. This method was validated extensively using bovine bone, and the effect of span/depth aspect ratio (5 vs 6) on the measured flexural properties was examined. The method provided reasonable results for both Young's modulus and flexural strength in bovine bone. With a span/depth ratio of 6, the median longitudinal modulus and flexural strength results were 16.1 (range: 14.4-19.3)GPa and 251 (range: 219-293)MPa, respectively. Finally, the pilot results from two osteotomy specimens from children with osteogenesis imperfecta are presented. These results provide the first measures of bone material strength in this patient population.

MicroRNA manipulation in colorectal cancer cells: from laboratory to clinical application.

The development of Colorectal Cancer (CRC) follows a sequential progression from adenoma to the carcinoma. Therefore, opportunities exist to interfere with the natural course of disease development and progression. Dysregulation of microRNAs (miRNAs) in cancer cells indirectly results in higher levels of messenger RNA (mRNA) specific to tumour promoter genes or tumour suppressor genes. This narrative review aims to provide a comprehensive review of the literature about the manipulation of oncogenic or tumour suppressor miRNAs in colorectal cancer cells for the purpose of development of anticancer therapies. A literature search identified studies describing manipulation of miRNAs in colorectal cancer cells in vivo and in vitro. Studies were also included to provide an update on the role of miRNAs in CRC development, progression and diagnosis. Strategy based on restoration of silenced miRNAs or inhibition of over expressed miRNAs has opened a new area of research in cancer therapy. In this review article different techniques for miRNA manipulation are reviewed and their utility for colorectal cancer therapy has been discussed in detail. Restoration of normal equilibrium for cancer related miRNAs can result in inhibition of tumour growth, apoptosis, blocking of invasion, angiogenesis and metastasis. Furthermore, drug resistant cancer cells can be turned into drug sensitive cells on alteration of specific miRNAs in cancer cells. MiRNA modulation in cancer cells holds great potential to replace current anticancer therapies. However, further work is needed on tissue specific delivery systems and strategies to avoid side effects.

Establishing community partnerships to support late-life anxiety research: lessons learned from the Calmer Life project.

OBJECTIVES: This article outlines the development of the Calmer Life project, a partnership established between researchers and faith-based and social service organizations to examine the effectiveness of cognitive behavioral therapy (CBT) incorporating religious/spiritual components for older African Americans in low-income communities. METHOD: The program was designed to bypass several barriers to delivery of CBT within the specified community; it allows multimodal delivery (in person or by telephone) that occurs outside traditional mental health settings through faith-based organizations and neighborhood community centers. It includes religion/spirituality as an element, dependent upon the preference of the participant, and is modular, so that people can select the skills they wish to learn. Established relationships within the community were built upon, and initial meetings were held in community settings, allowing feedback from community organizations. RESULTS: This ongoing program is functioning successfully and has strengthened relationships with community partners and facilitated increased availability of education and services in the community. The lessons learned in establishing these partnerships are outlined. CONCLUSIONS: The growth of effectiveness research for late-life anxiety treatments in underserved minority populations requires development of functional partnerships between academic institutions and community stakeholders, along with treatment modifications to effectively address barriers faced by these consumers. The Calmer Life project may serve as a model.

Chromatographic analysis of oxidized cello-oligomers generated by lytic polysaccharide monooxygenases using dual electrolytic eluent generation.

Research on oligosaccharides, including the complicated product mixtures generated by lytic polysaccharide monooxygenases (LPMOs), is growing at a rapid pace. LPMOs are gaining major interest, and the ability to efficiently and accurately separate and quantify their native and oxidized products chromatographically is essential in furthering our understanding of these oxidative enzymes. Here we present a novel set of methods based on dual electrolytic eluent generation, where the conventional sodium acetate/sodium hydroxide (NaOAc/NaOH) eluents in high-performance anion-exchange chromatography (HPAEC) are replaced by electrolytically-generated potassium methane sulfonate/potassium hydroxide (KMSA/KOH). The new methods separate all compounds of interest within 24-45 min and with high sensitivity; limits of detection and quantification were in the range of 0.0001-0.0032 mM and 0.0002-0.0096 mM, respectively. In addition, an average of 3.5 times improvement in analytical CV was obtained. This chromatographic platform overcomes drawbacks associated with manual preparation of eluents and offers simplified operation and rapid method optimization, with increased precision for less abundant LPMO-derived products.

Successful Production and Ligninolytic Activity of a Bacterial Laccase, Lac51, Made in Nicotiana benthamiana via Transient Expression.

Giant panda could have bamboo as their exclusive diet for about 2 million years because of the contribution of numerous enzymes produced by their gut bacteria, for instance laccases. Laccases are blue multi-copper oxidases that catalyze the oxidation of a broad spectrum of phenolic and aromatic compounds with water as the only byproduct. As a "green enzyme," laccases have potential in industrial applications, for example, when dealing with degradation of recalcitrant biopolymers, such as lignin. In the current study, a bacterial laccase, Lac51, originating from Pseudomonas putida and identified in the gut microbiome of the giant panda's gut was transiently expressed in the non-food plant Nicotiana benthamiana and characterized. Our results show that recombinant Lac51 exhibits bacterial laccase properties, with optimal pH and temperature at 7-8 and 40°C, respectively, when using syringaldazine as substrate. Moreover, we demonstrate the functional capability of the plant expressed Lac51 to oxidize lignin using selected lignin monomers that serve as substrates of Lac51. In summary, our study demonstrates the potential of green and non-food plants as a viable enzyme production platform for bacterial laccases. This result enriches our understanding of plant-made enzymes, as, to our knowledge, Lac51 is the first functional recombinant laccase produced in plants.

Electrochemical metallization memories--fundamentals, applications, prospects.

This review focuses on electrochemical metallization memory cells (ECM), highlighting their advantages as the next generation memories. In a brief introduction, the basic switching mechanism of ECM cells is described and the historical development is sketched. In a second part, the full spectra of materials and material combinations used for memory device prototypes and for dedicated studies are presented. In a third part, the specific thermodynamics and kinetics of nanosized electrochemical cells are described. The overlapping of the space charge layers is found to be most relevant for the cell properties at rest. The major factors determining the functionality of the ECM cells are the electrode reaction and the transport kinetics. Depending on electrode and/or electrolyte material electron transfer, electro-crystallization or slow diffusion under strong electric fields can be rate determining. In the fourth part, the major device characteristics of ECM cells are explained. Emphasis is placed on switching speed, forming and SET/RESET voltage, R(ON) to R(OFF) ratio, endurance and retention, and scaling potentials. In the last part, circuit design aspects of ECM arrays are discussed, including the pros and cons of active and passive arrays. In the case of passive arrays, the fundamental sneak path problem is described and as well as a possible solution by two anti-serial (complementary) interconnected resistive switches per cell. Furthermore, the prospects of ECM with regard to further scalability and the ability for multi-bit data storage are addressed.

Older adults' preferences for religion/spirituality in treatment for anxiety and depression.

OBJECTIVES: To examine patient preferences for incorporating religion and/or spirituality into therapy for anxiety or depression and examine the relations between patient preferences and religious and spiritual coping styles, beliefs and behaviors. METHOD: Participants (66 adults, 55 years or older, from earlier studies of cognitive-behavioral therapy for late-life anxiety and/or depression in primary care) completed these measures by telephone or in-person: Geriatric Anxiety Inventory, Client Attitudes Toward Spirituality in Therapy, Patient Interview, Brief Religious Coping, Religious Problem Solving Scale, Santa Clara Strength of Religious Faith, and Brief Multidimensional Measure of Religiousness and Spirituality. Spearman's rank-order correlations and ordinal logistic regression examined religious/spiritual variables as predictors of preferences for inclusion of religion or spirituality into counseling. RESULTS: Most participants (77-83%) preferred including religion and/or spirituality in therapy for anxiety and depression. Participants who thought it was important to include religion or spirituality in therapy reported more positive religious-based coping, greater strength of religious faith, and greater collaborative and less self-directed problem-solving styles than participants who did not think it was important. CONCLUSION: For individuals like most participants in this study (Christians), incorporating spirituality/religion into counseling for anxiety and depression was desirable.

Threat-related processing supports prospective memory retrieval for people with obsessive tendencies.

Obsessive-compulsive disorder can result in a variety of deficits to cognitive performance, including negative consequences for attention and memory performance. The question addressed in the current study concerned whether this disorder influenced performance in an event-based prospective memory task. The results from a subclinical population indicated that, relative to non-anxious controls and mildly depressed controls, people with obsessive-compulsive tendencies (washing compulsions) incur decrements in remembering to respond to cues related to a neutral intention (respond to animals). This deficit was ameliorated by giving the subclinical group an intention about a threat-related category (respond to bodily fluids) and cueing them with concepts that they had previously rated as particularly disturbing to them. Thus, their normal attentional bias for extended processing of threat-related information overcame their natural deficit in event-based prospective memory.

Empirically supported treatments in rural community mental health centers: a preliminary report on current utilization and attitudes toward adoption.

Rural community mental health centers (CMHCs) face numerous problems that might be alleviated by the dissemination of empirically supported treatments (ESTs). The current study lays the groundwork for EST dissemination by examining current treatment practices in rural clinics as well as the attitudes of decision makers toward ESTs and perceived barriers to their adoption. Twenty-five rural and 38 non-rural clinical directors responded to a mailed survey. Rural respondents were as likely as non-rural respondents to report EST use in their clinic for most anxiety disorders, and more likely to report use of an EST for major depressive disorder and obsessive-compulsive disorder. However, ESTs represent a relatively small proportion of the treatments reported for depression and anxiety disorders. Attitudes toward manualized ESTs did not differ between groups. Further, rural and nonrural clinics did not see the barriers to the adoption of ESTs as insurmountable. These preliminary results suggest that rural CMHCs are open to the use of ESTs and should be included in widespread dissemination initiatives.

If we build it, they might come: an empirical investigation of supply and demand in the recruitment of rural psychologists.

Past research has established that practicing doctoral level psychologists tend to cluster in metropolitan areas, leaving the rural population gravely underserved. Discussions of this problem have assumed that psychologists hold negative attitudes toward rural work for many reasons, leading to a supply shortage. The present study attempts to examine the accuracy of this hypothesis by examining attitudes of current doctoral students in clinical psychology. Eight hundred eighty-four students responded to an online survey. Results indicate that attitudes toward rural practice were not overwhelmingly negative as predicted. Additionally, a significant number of respondents endorsed positive attitudes toward rural practice. A follow-up study was conducted to determine if the shortage of rural psychologists might be attributed to a lack of demand from rural employers. A search of three popular job resources revealed that few rural jobs were advertised, indicating weak demand for psychologists in rural areas. Results indicate that efforts to recruit psychologists to rural areas should be rethought, and increasing the visibility of available opportunities for psychologists should be made a priority.

Neuroinspired unsupervised learning and pruning with subquantum CBRAM arrays.

Resistive RAM crossbar arrays offer an attractive solution to minimize off-chip data transfer and parallelize on-chip computations for neural networks. Here, we report a hardware/software co-design approach based on low energy subquantum conductive bridging RAM (CBRAM®) devices and a network pruning technique to reduce network level energy consumption. First, we demonstrate low energy subquantum CBRAM devices exhibiting gradual switching characteristics important for implementing weight updates in hardware during unsupervised learning. Then we develop a network pruning algorithm that can be employed during training, different from previous network pruning approaches applied for inference only. Using a 512 kbit subquantum CBRAM array, we experimentally demonstrate high recognition accuracy on the MNIST dataset for digital implementation of unsupervised learning. Our hardware/software co-design approach can pave the way towards resistive memory based neuro-inspired systems that can autonomously learn and process information in power-limited settings.

Functional characterization of a lytic polysaccharide monooxygenase from the thermophilic fungus Myceliophthora thermophila.

Thermophilic fungi are a promising source of thermostable enzymes able to hydrolytically or oxidatively degrade plant cell wall components. Among these enzymes are lytic polysaccharide monooxygenases (LPMOs), enzymes capable of enhancing biomass hydrolysis through an oxidative mechanism. Myceliophthora thermophila (synonym Sporotrichum thermophile), an Ascomycete fungus, expresses and secretes over a dozen different LPMOs. In this study, we report the overexpression and biochemical study of a previously uncharacterized LPMO (MtLPMO9J) from M. thermophila M77 in Aspergillus nidulans. MtLPMO9J is a single-domain LPMO and has 63% sequence similarity with the catalytic domain of NcLPMO9C from Neurospora crassa. Biochemical characterization of MtLPMO9J revealed that it performs C4-oxidation and is active against cellulose, soluble cello-oligosaccharides and xyloglucan. Moreover, biophysical studies showed that MtLPMO9J is structurally stable at pH above 5 and at temperatures up to 50°C. Importantly, LC-MS analysis of the peptides after tryptic digestion of the recombinantly produced protein revealed not only the correct processing of the signal peptide and methylation of the N-terminal histidine, but also partial autoxidation of the catalytic center. This shows that redox conditions need to be controlled, not only during LPMO reactions but also during protein production, to protect LPMOs from oxidative damage.

Life Values as an Intrinsic Guide for Cardiopulmonary Rehabilitation Program Engagement: A QUALITATIVE ANALYSIS.

PURPOSE: Participation in cardiovascular and pulmonary rehabilitation (CVPR) programs can lead to improved functional abilities and improved quality of life, but attendance and adherence to these programs remain suboptimal. Behavioral therapies have emphasized the importance of life value identification as a guide for goal setting and behavior change for both psychological and physical health conditions. Individuals who choose to engage in behaviors that align with their life values are thought to be intrinsically reinforced. The purpose of the following qualitative study was to interview patients enrolled in CVPR about their own life values and motivating factors related to healthy behavior changes. METHODS: Thirty cardiac or pulmonary patients were recruited from a CVPR program and participated in a semistructured interview about their life values and motivating factors related to program engagement. The data were transcribed and analyzed thematically. RESULTS: Participants identified a wide range of values related to program engagement, and only half of the participants endorsed health as a value. The most frequently endorsed life values included being active, family, and independence. The interviews indicated that, although patients make lifestyle changes in the program to improve their physical health, there are often other values that primarily guide their choice to engage in and maintain lifestyle behaviors. CONCLUSIONS: Life values can serve as a powerful guide for individual behavior change. The present study suggests that the piloting of brief values interventions early in CVPR treatment is warranted and has the potential to improve patient outcomes.

Classification of micro-CT images using 3D characterization of bone canal patterns in human osteogenesis imperfecta.

Few studies have analyzed the microstructural properties of bone in cases of Osteogenenis Imperfecta (OI), or 'brittle bone disease'. Current approaches mainly focus on bone mineral density measurements as an indirect indicator of bone strength and quality. It has been shown that bone strength would depend not only on composition but also structural organization. This study aims to characterize 3D structure of the cortical bone in high-resolution micro CT images. A total of 40 bone fragments from 28 subjects (13 with OI and 15 healthy controls) were imaged using micro tomography using a synchrotron light source (SRµCT). Minkowski functionals - volume, surface, curvature, and Euler characteristics - describing the topological organization of the bone were computed from the images. The features were used in a machine learning task to classify between healthy and OI bone. The best classification performance (mean AUC - 0.96) was achieved with a combined 4-dimensional feature of all Minkowski functionals. Individually, the best feature performance was seen using curvature (mean AUC - 0.85), which characterizes the edges within a binary object. These results show that quantitative analysis of cortical bone microstructure, in a computer-aided diagnostics framework, can be used to distinguish between healthy and OI bone with high accuracy.

The "two-week wait" referral pathway is not associated with improved survival for patients with colorectal cancer.

AIM: To improve survival rates in patients diagnosed with cancer in the UK, a two-week wait (2ww) referral to first appointment target and a 62 day referral to treatment target were introduced in 2004. This study analyses survival rates for patients diagnosed with colorectal cancer (CRC) by mode of referral and referral to treatment time. METHOD: A prospectively maintained database of CRC outcomes at the University Hospitals of Leicester NHS Trust was analysed. Data for patients diagnosed with CRC was analysed for survival. Comparisons were made by mode of referral (2ww, urgent, routine, emergency, national bowel cancer screening programme (NBCSP) and other screening pathways). In addition, this study assessed referral to initial treatment times for patients undergoing cancer resection (<62days group vs. >62days group). Inter-group comparisons were made using the Mann-Whitney-U-test. Kaplan-Meier survival probability estimates were calculated for overall survival and the log-rank test was used to compare the survival distributions in different groups. RESULTS: Overall survival (median time) was significantly lower for patients referred by the '2ww' pathway (3.5 years, 95% CI: 2.7-4.30), in comparison to the 'routine' (5.4 years, 95% CI: 4.5-6.6) pathway (p < 0.001). Patients referred on the '2ww' pathway were 1.34 times more likely to have stage IV disease at presentation in comparison to patients referred by the 'routine' pathway. Comparison of referral to initial treatment times showed there was no significant difference in survival between the <62days group and the >62days group (7.1 vs. 6.54, p = 0.620). CONCLUSION: Patients diagnosed with CRC by the 2ww pathway had shorter survival times than those referred by a routine pathway.