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The present study is aimed to identify the effect of gratitude as an adaptive regulating mechanism from suicidal ideation (SI) for veterans with mental illness (study 1) and student veterans with posttraumatic stress disorder (PTSD) symptoms (study 2) in the United States. Descriptive statistics and regression analyses were used to examine sociodemographic characteristics and relationships between gratitude and SI. Our study 1 consisted of 156 veterans with mental illness. The mean age for study 1 was 37.85. Our study 2 consisted of 232 student veterans with PTSD symptoms. The mean age for study 2 was 28.43. Higher gratitude scores in study 1 and study 2 were significantly associated with lower SI scores after adjusting for demographics and depression. This study partially supports the association between gratitude and SI in veterans with mental illness. Based on the results from this study, gratitude interventions may be effective in reducing SI when working with veterans with mental illness.

The cockroach allergen Bla g 1 forms a novel fold consisting of 12 amphipathic alpha-helices enclosing an exceptionally large hydrophobic cavity which was previously demonstrated to bind a variety of lipids. Since lipid-dependent immunoactivity is observed in numerous allergens, understanding the structural basis of this interaction could yield insights into the molecular determinants of allergenicity. Here, we report atomic modelling of Bla g 1 bound to both fatty-acid and phospholipids ligands, with 8 acyl chains suggested to represent full stoichiometric binding. This unusually high occupancy was verified experimentally, though both modelling and circular dichroism indicate that the general alpha-helical structure is maintained regardless of cargo loading. Fatty-acid cargoes significantly enhanced thermostability while inhibiting cleavage by cathepsin S, an endosomal protease essential for antigen processing and presentation; the latter of which was found to correlate to a decreased production of known T-cell epitopes. Both effects were strongly dependent on acyl chain length, with 18–20 carbons providing the maximal increase in melting temperature (~20 °C) while completely abolishing proteolysis. Diacyl chain cargoes provided similar enhancements to thermostability, but yielded reduced levels of proteolytic resistance. This study describes how the biophysical properties of Bla g 1 ligand binding and digestion may relate to antigen processing, with potential downstream implications for immunogenicity.

The mosquito protein AEG12 is up-regulated in response to blood meals and flavivirus infection though its function remained elusive. Here, we determine the three-dimensional structure of AEG12 and describe the binding specificity of acyl-chain ligands within its large central hydrophobic cavity. We show that AEG12 displays hemolytic and cytolytic activity by selectively delivering unsaturated fatty acid cargoes into phosphatidylcholine-rich lipid bilayers. This property of AEG12 also enables it to inhibit replication of enveloped viruses such as Dengue and Zika viruses at low micromolar concentrations. Weaker inhibition was observed against more distantly related coronaviruses and lentivirus, while no inhibition was observed against the nonenveloped virus adeno-associated virus. Together, our results uncover the mechanistic understanding of AEG12 function and provide the necessary implications for its use as a broad-spectrum therapeutic against cellular and viral targets.

Interleukins and neurotrophins levels are altered in the periphery of patients with major depression and suicidal behavior, however it is not clear if similar abnormalities occur in the central nervous system. Our objective was to examine the association of IL-6, IL-1β, BDNF, and GDNF levels between postmortem plasma, cerebrospinal fluid (CSF), and brain tissue in a heterogeneous diagnostic subject groups including normal controls, mood disorders only, mood disorders with AUD/SUD (alcohol abuse disorder, substance abuse disorder), and AUD/SUD without mood disorders. To address these questions we collected postmortem plasma (n = 29), CSF (n = 28), and brain (BA10) (n = 57) samples from individuals with mood disorder, mood disorder with AUD/SUD, AUD/SUD and normal controls. These samples were analyzed using a multiplex based luminex assay with a customized 4-plex cytokine/interleukins- IL-6, IL-1β, BDNF, and GDNF human acute phase based on xMAP technology platform. Protein levels were determined using a Luminex 200 instrument equipped with Xponent-analyzing software. We observed IL-6 (p = 2.1e-07), and GDNF (p = 0.046) were significantly correlated between brain and CSF. In addition, IL-6 (p = 0.031), were significantly correlated between brain and plasma. Overall diagnostic group analysis showed a significant difference with brain GDNF, p = 0.0106. Pairwise comparisons showed that GDNF level is—39.9 ± 12 pg/ml, p = 0.0106, was significantly higher than in the brains derived from mood disorders compared to normal controls, —23.8 ± 5.5 pg/ml, p = 0.034. Brain BDNF was higher in suicide (p = 0.0023), males compared to females (p = 0.017), and psychiatric medication treated vs. non-treated (p = 0.005) individuals. Overall, we demonstrate that blood IL-6, GDNF and BDNF could be informative peripheral biomarkers of brain biology associated with mood disorders, substance disorders, and suicide.

We present the final design of the Giant Magellan Telescope (GMT) Adaptive Secondary Mirrors System (ASMS), which comprises seven 1m class deformable mirrors segments plus seven hexapod positioners. Each deformable mirror is based on the well established contactless technology developed by AdOptica and already successfully deployed in several 8m class telescopes. The challenge for GMT is that the seven deformable mirrors will function as a single mirror. A subscale prototype made of 72 actuators has been produced to secure system final design: test setup and preliminary results are presented.

The Giant Magellan Telescope will be a 25.4-m visible and infrared telescope at Las Campanas Observatory. The optical design consists of 7 8.4-m primary mirror segments that reflect light to 7 secondary mirror segments in a doubly-segmented direct Gregorian configuration. Each mirror pair must be coaligned and co-boresighted. During operations, the alignment of the optical components will deflect due to variations in temperature, gravity-induced structure flexure of the mount, and, on a scale relevant to phasing, vibrations. The doubly-segmented nature and size of the GMT will create a novel set of challenges for initial assembly, integration, and verification and maintaining high-precision alignment of the optical elements during operations. GMT is developing a Telescope Metrology System that uses 3D laser metrology systems to decrease the complexity of alignment and increase observatory efficiency. This paper discusses the 4 subsystems of TMS as well as their operational modes.

Suicidality is one of the leading causes of death among young adults in the United States and represents a significant health problem worldwide. The suicide rate among adolescents in the United States has increased dramatically in the latest years and has been accompanied by considerable changes in youth suicide, especially among young girls. Henceforth, we need a good understanding of the risk factors contributing to suicidal behavior in youth. An explanatory model for suicidal behavior that links clinical and psychological risk factors to the underlying neurobiological, neuropsychological abnormalities related to suicidal behavior might predict to help identify treatment options and have empirical value. Our explanatory model proposes that developmental, biological factors (genetics, proteomics, epigenetics, immunological) and psychological or clinical (childhood adversities) may have causal relevance to the changes associated with suicidal behavior. In this way, our model integrates findings from several perspectives in suicidality and attempts to explain the relationship between various neurobiological, genetic, and clinical observations in suicide research, offering a comprehensive hypothesis to facilitate understanding of this complex outcome. Unraveling the knowledge of the complex interplay of psychological, biological, sociobiological, and clinical risk factors is highly essential, concerning the development of effective prevention strategy plans for suicidal ideation and suicide., Highlights • Suicidality, suicidal attempts in adolescents is currently one of the major concern in public and mental health issues worldwide. • Positive and Negative symptoms of suicide in adolescents. • Neuropsychology, neurobiology- neurochemical, neuroimmunological and sociobiological risk factors of suicidality in adolescents. • Early risk assessments and its impact on treatment strategies for suicide.

OBJECTIVES: Recently fructose has been linked to the development of Nonalcoholic Fatty Liver Disease (NAFLD), but the mechanisms behind the progression remain to be elucidated. Kupffer cells have been identified as regulators of hepatic inflammation and extracellular matrix (ECM) proteins. Activated macrophages are known to express tissue inhibitor of metalloproteinase (TIMP1), which inhibits matrix metalloproteinase 9 (MMP9) activity of reconstructing ECM which leads to fibrosis. In humans and mouse models, TIMP1 expression is positively correlated with the progression of NAFLD. Based on these findings, we hypothesize that fructose regulates TIMP1 gene expression in activated proinflammatory macrophages. METHODS: Using an in vitro model of J774 macrophages and primary Kupffer cells, cells were treated to induce an M1 phenotype in the presence of glucose or fructose. Cells were harvested for RNA and RT-PCR was conducted to measure ECM gene expression. Isolated Kupffer cells were collected from C57BL/6J mice fed a high fat diet (HFD) supplemented with 30% fructose or 30% glucose and analyzed for ECM expression. To examine fructose uptake and intra- and extracellular metabolites, mass spectrophotometry and nuclear magnetic resonance was conducted. RESULTS: Timp1 gene expression was significantly increased in J77.4 cells treated with fructose compared to glucose. Surprisingly, fructose treatment decreased Mmp9 gene expression. Likewise, fructose treatment increased TIMP1 protein expression in isolated Kupffer cells. In vivo, isolated hepatic macrophages from mice fed HF and high fructose diet had elevated Timp1 gene expression compared to mice fed high glucose diet. Extracellular levels of lactate decreased by 1.5-fold in fructose treated J77.4 cells compared to glucose. Metabolites involved in the TCA cycle and mitochondrial function were decreased when treated with fructose compared to glucose in non-activated macrophages. However, fructose treatment increased intracellular methanol and acetate levels in M1 macrophages compared to glucose. CONCLUSIONS: Our data suggest that fructose upregulates Timp1 expression and possibly decreases mitochondrial function while increasing acetate and methanol production, identifying new mechanisms by which fructose drives the progression of NAFLD. FUNDING SOURCES: Kenan Institute North Carolina University.

The daily pollen forecast provides crucial information for allergic patients to avoid exposure to specific pollen. Pollen counts are typically measured with air samplers and analyzed with microscopy by trained experts. In contrast, this study evaluated the effectiveness of identifying the component pollens using the metabolites extracted from an air-sampled pollen mixture. Ambient air-sampled pollen from Munich in 2016 and 2017 was visually identified from reference pollens and extracts were prepared. The extracts were lyophilized, rehydrated in optimal NMR buffers, and filtered to remove large proteins. NMR spectra were analyzed for pollen associated metabolites. Regression and decision-tree based algorithms using the concentration of metabolites calculated from the NMR spectra outperformed algorithms using the NMR spectra themselves as input data for pollen identification. Categorical prediction algorithms trained for low, medium, high, and very high pollen count groups had accuracies of 74% for the tree, 82% for the grass, and 93% for the weed pollen count. Deep learning models using convolutional neural networks performed better than regression models using NMR spectral input, and were the overall best method in terms of relative error and classification accuracy (86% for tree, 89% for grass, and 93% for weed pollen count). This study demonstrates that NMR spectra of air-sampled pollen extracts can be used in an automated fashion to provide taxa and type-specific measures of the daily pollen count.

Background Phosphatidylethanol (PEth) is a metabolite of ethanol (EtOH), and its concentration in whole blood samples is a direct biomarker of alcohol consumption. Because PEth is also present in the brain and incorporated in lipid membranes, it can be used to classify deceased individuals on alcohol consumption status at the time of death. The purpose of this study was to detect PEth homologs in postmortem brains of individuals known to have had alcohol use disorder (AUD) and to determine the relationship between serum alcohol at the time of death and PEth in the cerebellum (CE) and orbital frontal cortex (OFC). Methods Postmortem brain was collected and stored according to standard protocol. Psychiatric symptoms experienced prior to death were obtained by next of kin psychological autopsy to categorize subjects. Thirty male subjects were chosen for analyses: 10 with AUD with positive serum EtOH levels present at time of autopsy (AUD-W), 10 with AUD without positive serum EtOH levels (AUD-WO), and 10 controls. PEth 16:0/18:1 and 16:0/18:2 were quantified in 50 mg of CE and OFC of human postmortem brain using HPLC and mass spectrometric detection (triple quadrupole). Results Results of this study were as follows: (i) PEth 16:0/18:1 and 16:0/18:2 were detected in the CE and OFC of all subjects diagnosed with AUD, (ii) PEth 16:0/18:1 levels were about 10-fold higher than PEth 16:0/18:2 in all subjects and both areas of brain, (iii) AUD-W subjects had higher PEth homolog levels in CE and OFC than controls and AUD-WO subjects, (iv) PEth 16:0/18:1, but not PEth 16:0/18:2, levels in CE and OFC of AUD-W subjects correlated significantly with serum EtOH levels at the time of death. Conclusions Quantification of combined PEth homolog levels in postmortem human brain is a good candidate as a diagnostic factor to classify drinking status, especially for those with AUD at the time of death. For alcohol research studies with postmortem brain, verification of drinking status is essential.

Vinculin is an essential adhesion protein that links membrane-bound integrin and cadherin receptors through their intracellular binding partners to filamentous actin, facilitating mechanotransduction. Here we present an 8.5-Å-resolution cryo-electron microscopy reconstruction and pseudo-atomic model of the vinculin tail (Vt) domain bound to F-actin. Upon actin engagement, the N-terminal “strap” and helix 1 are displaced from the Vt helical bundle to mediate actin bundling. We find that an analogous conformational change also occurs in the H1′ helix of the tail domain of metavinculin (MVt) upon actin binding, a muscle-specific splice isoform that suppresses actin bundling by Vt. These data support a model in which metavinculin tunes the actin bundling activity of vinculin in a tissue-specific manner, providing a mechanistic framework for understanding metavinculin mutations associated with hereditary cardiomyopathies.

Launched in 2009, Keck Observatory’s Telescope Control System Upgrade (TCSU) project set out to improve Keck’s telescope pointing, tracking, and offsetting performance as well as increase maintainability and reliability. The project went online full time on the Keck 2 telescope in October 2017 and on the Keck 1 telescope in March 2018 after a notable delay due to a re-design of the azimuth and elevation encoder mounting systems. This paper discusses the details and challenges of implementing this large and complicated system while never requiring a shutdown of either telescope. The TCSU project replaced all of the major elements of the telescope controls, rotator and secondary mirror controls, and safety system. National Instrument’s reconfigurable I/O technology (i.e. NI RIO), with their embedded field programmable gate arrays (FPGAs), are used as the core of the telescope’s digital velocity control loop, structural filter, and tachometer filter. They were also used to create a monitoring and safety system for the rotator velocity controller as well as reading the newly installed tilt meters used to greatly improve pointing performance. Delta Tau’s family of “Brick” programmable multi-axis controllers, i.e. PMAC or BRICK, are used to control the rotator and secondary mirror. They enable better tuning and faster slew speeds for these subsystems. An Allen Bradley’s ControlLogix® controller and the family of FLEX™ Input/Output (IO) modules were used to create a distributed safety system able to handle a wide variety of signal types. This technology refresh based on commercial off the shelf equipment replaced much of our obsolete and custom equipment. A significant part of the project was the installation of new telescope azimuth and elevation position encoders based on Heidenhain’s 40 micron grading tape scales. Interpolated to a 10 nanometer resolution, the new encoders provide true 4 mas resolution in azimuth and 1 mas resolution in elevation. This is a big improvement to Keck’s position sensing when compared to the old rotary incremental encoders. The installation required a significant amount of mechanical infrastructure to house them. Additionally, two tilt meters were installed to sense the telescope’s varying vertical angle as a function of azimuth, mainly due to the azimuth bearing’s axial runout. The encoders and tilt meters are the primary reason for achieving the greatly improved pointing and tracking performance [1]. Finally, a switching solution using solid state relays and dual network switches was installed to provide seamless and rapid switching between the old and new control systems during commissioning. Although this component is a simple design and does not boast of any new technology, it is one of the key components that enabled the successful testing of the new equipment while keeping the old system operational as a backup for night time observing as well as for baseline performance comparisons. It allowed us to switch a variety of signal types and was very cost effective when compared to available products.

Synaptosomal Associated Protein-25 kilodaltons (SNAP-25) is an integral member of the SNARE complex. This complex is essential for calcium-triggered synaptic vesicular fusion and release of neurotransmitters into the synaptic cleft. In addition to neurotransmission, SNAP-25 is associated with insulin release, the regulation of intracellular calcium, and neuroplasticity. Because of SNAP-25's varied and crucial biological roles, the consequences of changes in this protein can be seen in both the central nervous system and the periphery. In this review, we will look at the published literature from human genetic, postmortem, and animal studies involving SNAP-25. The accumulated data indicate that SNAP-25 may be linked with some symptoms associated with a variety of psychiatric disorders. These disorders include bipolar disorder, schizophrenia, major depressive disorder, attention deficit hyperactivity disorder, autism, alcohol use disorder, and dementia. There are also data suggesting SNAP-25 may be involved with non-psychiatric seizures and metabolic disorders. We believe investigation of SNAP-25 is important for understanding both normal behavior and some aspects of the pathophysiology of behavior seen with psychiatric disorders. The wealth of information from both animal and human studies on SNAP-25 offers an excellent opportunity to use a bi-directional research approach. Hypotheses generated from genetically manipulated mice can be directly tested in human postmortem tissue, and, conversely, human genetic and postmortem findings can improve and validate animal models for psychiatric disorders.

The Giant Magellan Telescope (GMT) is an advanced Extremely Large Telescope and is being designed for broad wavelength coverage and maximum sensitivity in pursuit of several high priority cosmology science objectives. The GMT will use large primary mirror segments to provide well corrected wavefronts and a very fast optical system with a 20-arcminute diameter field of view. It will have a direct Gregorian optical prescription and deformable secondary mirrors to implement adaptive optics. The project timeline leads to initial commissioning with early instruments by 2023. The Systems Engineering group, along with the chief engineer, is developing and flowing down science and operational requirements and is the guardian of the performance budget. An integrated performance model of the telescope is being developed to support requirements derivation, performance assessment and trade studies that drive telescope architecture decisions. This model has four key elements; a structural plant model which exhibits the complex dynamics of the telescope; a controls model that models both controller dynamics and noise; an optics model to accurately demonstrate performance and optical feedback; and loads models to characterize various environmental loads. This paper will introduce the GMT integrated modeling architecture, but will focus on the specific tools and process developed to build the critical structural model. An efficient process was required to support the variety of required analyses, update frequency of underlying finite element models, and various project critical trade studies. The end-to-end process, from subsystem finite element models to a system level plant and Simulink block diagram has been achieved through development of a new MATLAB class with many useful functions. Functions are covered that combine the subsystem models, manipulate those models, and help them run efficiently in discrete and continuous time. Verification and version control are also discussed. Case study of wind is presented with preliminary performance results.

Aims The enzyme catechol-O-methyl transferase (COMT) plays a primary role in the metabolism of catecholamine neurotransmitters and is implicated in the modulation of cognitive and emotional responses. The best-characterized single nucleotide polymorphism (SNP) of the COMT gene consists of a valine (Val)-to-methionine (Met) substitution at codon 108/158. The Met-containing variant confers a marked reduction in COMT catalytic activity. We recently showed that the activity of recombinant COMT is positively regulated by the enzyme Met sulfoxide reductase (MSR), which counters the oxidation of Met residues of proteins. The current study was designed to assess whether brain COMT activity may be correlated to MSR in an allele-dependent fashion.

SNAP-25 is a neurotransmitter vesicular docking protein which has been associated with brain disorders such as attention deficit hyperactivity disorder, bipolar disorder and schizophrenia. In this project, we were interested if clinical factors are associated with differential SNAP-25 expression. We examined the SNAP-25 isoform mRNA and protein levels in postmortem cortex Brodmann's area 9 (BA9) and BA24 (n = 29). Subjects were divided by psychiatric diagnosis, clinical variables including mood state in the last week of life and lifetime impulsiveness. We found affected subjects with a diagnosis of alcohol use disorder (AUD) had a lower level of SNAP-25b BA24 protein compared to those without AUD. Hispanic subjects had lower levels of SNAP-25a, b and BA9 mRNA than Anglo-American subjects. Subjects who smoked had a total pan (total) SNAP-25 BA9/BA24 ratio. Subjects in the group with a low level of anxious-psychotic symptoms had higher SNAP-25a BA24 mRNA compared to normal controls, and both the high and low symptoms groups had higher pan (total) SNAP-25 BA9/BA24 ratios than normal controls. These data expand our understanding of clinical factors associated with SNAP-25. They suggest that SNAP-25 total and isoform levels may be useful biomarkers beyond limited neurological and psychiatric diagnostic categories.

The Thirty Meter Telescope (TMT) needs to maintain precise positioning of the optical elements to deliver unprecedented image quality. Any vibration from observatory sources must therefore be minimized; model-based analysis leads to maximum allowable forces from any individual source typically of order a Newton or less in the most sensitive frequency band. Careful attention throughout the design process is needed to ensure that these challenging requirements are satisfied. We focus here in particular on cogging forces from the azimuth motor drive. Each motor yields periodic tangential and vertical forces of order 100's of Newtons, with higher harmonics of the waveform potentially exciting telescope structural resonances. There are 56 drive motors, and appropriate phasing between them can ideally cancel most of the net torque or net vertical force. However, the moment created between non-collocated forces still produces image motion even if the net force cancels, and further, small errors in forcer positioning result in imperfect cancellation. We provide a general methodology for estimating total cogging forces and then present an example of the expected TMT performance impact from motor cogging placed in context with the larger challenge of demanding vibration requirement.

Circadian abnormalities may be related to mood disorders. Circadian gene expression was measured in postmortem brain tissue from individuals with affective disorders and controls. Relationships between circadian gene expression, clinical characteristics, and alcohol and psychotropic medication use were noted. Further study is warranted to characterize these relationships.

Venlafaxine (VEN) and its metabolite O-desmethylvenlafaxine (ODV) inhibit reuptake of serotonin and norepinephrine. This study examines whether VEN is differentially distributed in postmortem brain and examines relationships between brain and femoral blood concentrations from donors prescribed VEN for treatment of depression. Using high-pressure liquid chromatography-ultraviolet detection, VEN and ODV concentrations were measured in temporal, occipital, and cerebellar cortex of six postmortem brains. The ODV/VEN ratio was calculated as a relative measure of drug metabolism within each region where higher ratios indicated a greater conversion of VEN to ODV. Compared to the other regions examined, the cerebellum showed decreased VEN (p = 0.056), ODV (p = 0.006), and ODV/VEN (p = 0.027) ratios. In parts per million, VEN was higher in temporal and occipital cortex, but not cerebellum, as compared to femoral blood concentration. These observations suggest that VEN and ODV are differentially distributed in the brain, and metabolism of VEN to ODV may vary across brain regions.

Current and planned large optical telescopes use a segmented primary mirror, with the out-of-plane degrees of freedom of each segment actively controlled. The primary mirror of the Thirty Meter Telescope (TMT) considered here is composed of 492 segments, with 1476 actuators and 2772 sensors. In addition to many more actuators and sensors than at existing telescopes, higher bandwidths are desired to partially compensate for wind-turbulence loads on the segments. Control-structure interaction (CSI) limits the achievable bandwidth of the control system. Robustness can be further limited by uncertainty in the interaction matrix that relates sensor response to segment motion. The control system robustness is analyzed here for the TMT design, but the concepts are applicable to any segmented-mirror design. The key insight is to analyze the structural interaction in a Zernike basis; rapid convergence with additional basis functions is obtained because the dynamic coupling is much stronger at low spatial-frequency than at high. This analysis approach is both computational efficient, and provides guidance for structural optimization to minimize CSI.

Studies on ragweed and birch pollen extracts suggested that the adenosine content is an important factor in allergic sensitization. However, exposure levels from other pollens and considerations of geographic and seasonal factors have not been evaluated.This study compared the metabolite profile of pollen species important for allergic disease, specifically measured the adenosine content, and evaluated exposure to pollen-derived adenosine.An NMR metabolomics approach was used to measure metabolite concentrations in twenty-six pollen extracts. Pollen count data was analyzed from five cities to model exposure.A principal component analysis of the various metabolites identified by NMR showed that pollen extracts could be differentiated primarily by sugar content: glucose, fructose, sucrose, and myo-inositol. In extracts of 10 mg of pollen/ml, the adenosine was highest for grasses (45 μM) followed by trees (23 μM) and weeds (19 μM). Pollen count data showed that tree pollen was typically 5-10 times the amount of other pollens. At the daily peaks of tree, grass, and weed season the pollen-derived adenosine exposure per day is likely to only be 1.1, 0.11, and 0.12 μg, respectively. Seasonal models of pollen exposure and respiration suggest that it would be a rare event limited to tree pollen season for concentrations of pollen-derived adenosine to approach physiological levels.Sugar content and other metabolites may be useful in classifying pollens. Unless other factors create localized exposures that are very different from these models, pollen-derived adenosine is unlikely to be a major factor in allergic sensitization.

Deletions in the 16.6 kb mitochondrial genome have been implicated in numerous disorders that often display muscular and/or neurological symptoms due to the high-energy demands of these tissues. We describe a catalogue of 4489 putative mitochondrial DNA (mtDNA) deletions, including their frequency and relative read rate, using a combinatorial approach of mitochondria-targeted PCR, next-generation sequencing, bioinformatics, post-hoc filtering, annotation, and validation steps. Our bioinformatics pipeline uses MapSplice, an RNA-seq splice junction detection algorithm, to detect and quantify mtDNA deletion breakpoints rather than mRNA splices. Analyses of 93 samples from postmortem brain and blood found (i) the 4977 bp ‘common deletion’ was neither the most frequent deletion nor the most abundant; (ii) brain contained significantly more deletions than blood; (iii) many high frequency deletions were previously reported in MitoBreak, suggesting they are present at low levels in metabolically active tissues and are not exclusive to individuals with diagnosed mitochondrial pathologies; (iv) many individual deletions (and cumulative metrics) had significant and positive correlations with age and (v) the highest deletion burdens were observed in major depressive disorder brain, at levels greater than Kearns–Sayre Syndrome muscle. Collectively, these data suggest the Splice-Break pipeline can detect and quantify mtDNA deletions at a high level of resolution.

We computationally designed a de novo protein-protein interaction between wild-type ubiquitin and a redesigned scaffold. Our strategy was to incorporate zinc at the designed interface to promote affinity and orientation specificity. A large set of monomeric scaffold surfaces were computationally engineered with three-residue zinc coordination sites, and the ubiquitin residue H68 was docked to the open coordination site to complete a tetrahedral zinc site. This single coordination bond was intended as a hotspot and polar interaction for ubiquitin binding, and surrounding residues on the scaffold were optimized primarily as hydrophobic residues using a rotamer-based sequence design protocol in Rosetta. From thousands of independent design simulations, four sequences were selected for experimental characterization. The best performing design, called Spelter, binds tightly to zinc (Kd

Nonlinearities in aeroservoelastic systems are a source of limit-cycle oscillations. Aerodynamic control surfaces and actuators may have excessive free play in connecting joints and hinges, which has the potential to cause limit-cycle oscillations associated with aeroservoelastic vibrations, impacting handling and ride quality and leading to structural fatigue. Because of these negative impacts, free-play limits have been established by the Joint Services Guidance Military Specification. These military specifications have also been adopted by the Federal Aviation Administration for commercial aircraft. This stringent requirement can be excessively conservative and is very difficult and costly to meet. Analytical tools using describing functions have been developed to establish free-play limits on aeroservoelastic aircraft systems without adverse consequences. These tools are robust and easy to implement, justifying relaxation or validation of the conservative requirements. It is shown that oscillation fre...

Vinculin functions as a molecular clutch that organizes leading edge F-actin, generates traction, and promotes focal adhesion formation and turnover but not adhesion growth., In migrating cells, integrin-based focal adhesions (FAs) assemble in protruding lamellipodia in association with rapid filamentous actin (F-actin) assembly and retrograde flow. How dynamic F-actin is coupled to FA is not known. We analyzed the role of vinculin in integrating F-actin and FA dynamics by vinculin gene disruption in primary fibroblasts. Vinculin slowed F-actin flow in maturing FA to establish a lamellipodium–lamellum border and generate high extracellular matrix (ECM) traction forces. In addition, vinculin promoted nascent FA formation and turnover in lamellipodia and inhibited the frequency and rate of FA maturation. Characterization of a vinculin point mutant that specifically disrupts F-actin binding showed that vinculin–F-actin interaction is critical for these functions. However, FA growth rate correlated with F-actin flow speed independently of vinculin. Thus, vinculin functions as a molecular clutch, organizing leading edge F-actin, generating ECM traction, and promoting FA formation and turnover, but vinculin is dispensible for FA growth.

Axon outgrowth during development and neurotransmitter release depends on exocytotic mechanisms, although what protein machinery is common to or differentiates these processes remains unclear. Here we show that the neural t-SNARE (target-membrane-associated-soluble N-ethylmaleimide fusion protein attachment protein (SNAP) receptor) SNAP-25 is not required for nerve growth or stimulus-independent neurotransmitter release, but is essential for evoked synaptic transmission at neuromuscular junctions and central synapses. These results demonstrate that the development of neurotransmission requires the recruitment of a specialized SNARE core complex to meet the demands of regulated exocytosis.

Background Minority populations have been under-represented in mental health research studies. The systematic treatment enhancement program for bipolar disorder developed the Community Partners Program (CPP) to address this issue in a large, prospective treatment study of persons with bipolar disorder. Purpose The primary goal of CPP was to develop a community-based infrastructure for studying bipolar disorder that would enhance the ethnic/racial and socioeconomic diversity of participants. Methods Selected academic sites partnered with local clinics (n = 6 partnerships in five cities). This report describes the conceptualization, implementation, and qualitative evaluation of CPP, as well as quantitative analysis of clinical and sociodemographic differences between the samples recruited at academic versus community sites. Results Quantitative analysis of the 155 participants from the six partnerships revealed enrollment of 45% from minority populations (vs. 15% in academic sites). Significant sociodemographic differences were evident not only between academic and community sites, but within minority and non-minority groups across site types. Notably, clinical differences were not evident between participants from academic and community sites. Review of qualitative data suggests that certain factors around implementation of research protocols may enhance community participation. Conclusions Moving research recruitment and participation into community sites was more successful in increasing minority enrollment than efforts to attract such individuals to academic sites. Recommendations for creating and maintaining academic/community partnerships are given. Limitations Several important variables were not considered including mood severity, hospitalization, or treatment differences. Minority participants were grouped by combining African American and Hispanics, which may have obscured subgroup differences. A derivation of standard qualitative methods was used in this study. Clinical Trials 2009; 6: 344—354. http://ctj.sagepub.com

The enzyme catechol-O-methyltransferase (COMT), which catalyses the degradation of dopamine and norepinephrine, is posited to participate in the pathophysiology of bipolar disorder (BD) and schizophrenia. In support of this notion, rich evidence has documented that the severity of various BD and schizophrenia symptoms is moderated by rs4680, a single nucleotide polymorphism of the COMT gene featuring a valine (Val)-to-methionine (Met) substitution that results in lower catalytic activity. Nevertheless, the specific relevance of COMT enzymatic activity in the pathophysiology of BD and schizophrenia dimensions remains elusive.We measured COMT catalytic activity in post-mortem prefrontal cortices, striata and cerebella of schizophrenia and BD patients, as well as non-affected controls. These values were then correlated with rs4680 genotypes and psychopathology scores in the last week of life.No direct correlation between COMT activity and rs4680 genotypes was found; however, the severity of manic symptoms was highly correlated with COMT activity in the striatum, irrespective of the diagnostic group.These results suggest that COMT striatal activity, but not rs4680 genotype, may serve as a biomarker for manic symptoms. Future studies are warranted to confirm these findings and assess the neurobiological links between COMT striatal activity and manic symptoms.

Vinculin is an essential and abundant cytoskeletal protein localized to focal adhesions and cell-cell contacts, where it participates in the linkage of transmembrane receptors to the actin cytoskeleton to control cell survival and migration. Loss of vinculin results in increased cell migration, apoptotic resistance, and the acquisition of tumorigenic properties. Mutations in vinculin and its splice variant, metavinculin, are associated with cardiac disease. Vinculin consists of a head domain (Vh) and a tail domain (Vt) that form autoinhibitory interactions in its inactive state, but release upon activation, exposing phosphorylation, protein and phosphoinositol 4,5-bisphosphate (PIP2) binding sites. The interaction of vinculin with PIP2 is believed critical for its function, however, it is currently unclear how vinculin specifically recognizes PIP2 and regulates vinculin. Vt forms an antiparallel five-helix bundle with amino-terminal (NT) and carboxyl terminal (CT) extensions. While a crystal structure of an oligomerized Vt mutant complexed to a short chain PIP2 has recently been published (Chinthalapudi et al. (JCB, 2014)), the structure is incompatible with membrane insertion. We propose an alternative model using experimental data, molecular docking and dynamics simulations and provide validation of the model through biophysical and biochemical analyses. In our model, the PIP2 head group binds to the Vt basic collar, and promotes release of Vt's strap and CT to facilitate membrane insertion. The role of vinculin/PIP2 interaction in mediating vinculin activation, localization, cell migration, force sensing and transmission has also been characterized using cell microscopy, including super-resolution microscopy approaches, by examining WT and PIP2-deficient full length vinculin in knockout cells. Information derived from these analyses will result in an unprecedented understanding of vinculin function from the molecular to the cellular level and will enable us to build more comprehensive models of vinculin membrane interactions.

Objective: We assessed the spectrum and severity of bipolar symptoms that differentiated bipolar disorder (BD) clinical states, employing the Bipolar Inventory of Symptoms Scale (BISS) which provides a broader item range of traditional depression and mania rating scales. We addressed symptoms differentiating mixed states from depression or mania/hypomania. Method: One hundred and sixteen subjects who met DSM-IV-TR criteria for BD and were currently in a depressed, manic/hypomanic, mixed episode, or recovered state were interviewed using the BISS. Results: A subset of manic items differed between mixed episodes and mania/hypomania or depression. Most anxiety items were more severe in mixed subjects. BISS Depression and Manic subscales differentiated episodes from recovered status. The majority of depression and manic symptoms differentiated mood states in the predicted direction. Mixed episodes had overall greater mood severity than manic/hypomanic episodes or depressed episodes. Conclusion: These results indicate that a small subset of symptoms, several of which are absent in DSM-IV-TR criteria and traditional rating scales for bipolar studies, aid in distinguishing mixed episodes from depressive or manic/hypomanic episodes. The results also support the utility of a comprehensive BD symptom scale in distinguishing primary clinical states of BD.

Bowden CL, Singh V, Weisler R, Thompson P, Chang X, Quinones M, Mintz J. Lamotrigine vs. lamotrigine plus divalproex in randomized, placebo-controlled maintenance treatment for bipolar depression. Objective: To compare the maintenance efficacy of lamotrigine (Lam) to combination therapy of Lam + divalproex ER (Div) in recently depressed patients with bipolar disorder (BD). Method: We randomized 86 BD I or II patients in a major depressive episode to 8 months of double-blind treatment with Lam + placebo or Lam + Div. To be eligible for randomization, patients had to achieve control of both depressive and manic symptoms during an open phase that included both Lam and Div. Results: Time to depressive episode did not differ significantly by Kaplan–Maier survival analysis (χ2 = 1.82, df = 1, P = 0.18). However, several secondary outcomes did show significant differences. The proportion of Lam + placebo patients who had at least one Montgomery–Asberg Depression Rating Scale (MADRS) score ≥15 during the maintenance phase was 67% (30/45) compared with 44% (18/41) for the Lam + Div group (χ2 = 4.51, P = 0.03). Among BD I patients assigned to Lam + placebo, 71.4% (25/35) had at least one visit with MADRS score ≥15 compared with 36.7% (11/30) among Lam + Div patients (χ2 = 7.89, df = 1, P = 0.005). Conclusion: Lam + Div generally provided greater maintenance efficacy than Lam alone for depressive indices in recently depressed BD patients.

Thompson PM, Cruz DA, Olukotun DY, Delgado PL. Serotonin receptor, SERT mRNA and correlations with symptoms in males with alcohol dependence and suicide. Objective: This study tested the hypothesis that abnormalities in components of the serotonin (5HT) system in the prefrontal cortex are associated with suicide in alcohol-dependent subjects. Second, we assessed the relationship of lifetime impulsivity and mood symptoms with prefrontal cortex 5-HT measures. Method: Tissue was obtained from Brodmann’s areas (BA) 9 and 24 in postmortem samples of individuals who were alcohol dependent with suicide (n = 5), alcohol dependent without suicide (n = 9) and normal controls (n = 5). Serotonin receptor (5HT) and serotonin reuptake transporter (SERT) mRNA were measured. Interviews with next of kin estimated lifetime impulsivity and mood symptoms in the last week of life. Results: Serotonin receptor 1A (5HT1A) mRNA in BA 9 was elevated in the alcohol dependence without suicide group compared with controls. In the alcohol dependence with suicide group, anxiety symptoms were associated with decreased BA 24 SERT mRNA and depressive symptoms with BA 9 5HT1A mRNA expression. In the alcohol dependent only group impulsivity is correlated with increased BA 9, and BA 24 serotonin receptor 2A mRNA. Conclusion: Our data suggest region-specific change, rather than global serotonin blunting is involved in alcohol dependence and suicide. It also suggests that symptoms are differentially influenced by prefrontal cortex serotonin receptor mRNA levels.

Flight testing for aeroservoelastic clearance is an expensive and time consuming process. Large degree-of-freedom high-fidelity nonlinear aircraft models using computational fluid dynamics coupled with finite element models can be used for accurately predicting aeroelastic phenomena in all flight regimes including subsonic, supersonic, and transonic. With the incorporation of an active feedback control system, these high-fidelity models can be used to reduce the flight-test time needed for aeroservoelastic clearance. Accurate computational fluid dynamics/finite element models are computationally complex, rendering their runtime ill suited for adequate flight control system design. In this work, a complex, large-degree-of-freedom, transonic, inviscid computational fluid dynamics/finite element model of a fighter aircraft is fitted with a flight control system for aeroelastic oscillation reduction. A linear reduced-order model of the complete aeroelastic aircraft dynamic system is produced directly from the high-order nonlinear computational fluid dynamics/finite element model. This rapid runtime reduced-order model is used for the design of the flight control system, which includes models of the actuators and common nonlinearities in the form of rate limiting and saturation. The oscillation reduction controller is successfully demonstrated via a simulated flight test using the high-fidelity nonlinear computational fluid dynamics/finite element/flight control system model.

Current symptom rating scales and diagnostic categories for bipolar disorder (BD) do not provide dimensional profiles of the types of behavior disturbed in this complex disorder. To overcome these limitations we identified the principal domains of behavioral symptomatology in bipolar individuals, including all mood states, and used a more comprehensive rating scale for BD: the Bipolar Inventory of Signs and Symptoms Scale (BISS). A total of 246 patients with BD (196 with BD type I, and 50 with BD type II) were interviewed using the BISS. Exploratory factor analysis was performed on the BISS results using the maximum likelihood factor extraction method, followed by oblique rotation of the extracted factor loadings. We determined the strength of relationships between factor scores using the Pearson correlation coefficient. The following five factors were extracted: mania, depression, irritability, anxiety and psychosis. Anxiety was significantly correlated with depression and irritability. The mania factor score was only weakly associated with the other four factors. The domains of the BISS capture both the historical categories of depression and mania, plus irritability, psychosis, and an additional principal domain, anxiety. Despite the common occurrence of anxiety in BD, it has not been identified in most prior factor analyses, in part due to limited coverage of anxiety symptoms in the source scales.

Scales used in studies of bipolar disorder have generally been standardized with major depressive or hospitalized manic patients. A clinician rated scale based on a semi-structured interview for persons with bipolar disorder, with comprehensive coverage of bipolar symptomatology, is needed. We report concurrent, divergent and convergent psychometric reliability, discriminant validity and relationship to a measure of overall function for a new psychometric rating instrument. A primarily outpatient sample of 224 subjects was assessed using the Bipolar Inventory of Symptoms Scale (BISS). The BISS total score and depression and mania subscales were compared to the Young Mania Rating Scale (YMRS), the Montgomery Asberg Depression Rating Scale (MADRS) and the Global Assessment of Functioning Scale (GAF). Clinical mood states were also compared using the BISS. The BISS scores demonstrated good concurrent validity, with estimates (Pearson correlations) ranging from 0.74 to 0.94 for YMRS and MADRS and test-retest reliability from 0.95 to 0.98. BISS concurrent validity with the GAF was significant for four clinical states, but not mixed states. The BISS discriminated primary bipolar mood states as well as subjects recovered for eight weeks compared to healthy controls. In conclusion, the BISS is a reliable and valid instrument broadly applicable in clinical research to assess the comprehensive domains of bipolar disorder. Future directions include factor analysis and sensitivity to change from treatment studies.

The linear quadratic Gaussian (LQG) optimal control problem of single-input-single-output (SISO) analogue and digital systems can be solved using Wiener-Hopf techniques of spectral and partial factorization. A new form of the Wiener-Hopf result is derived, followed by a description of how the solution is implemented in Program CC, a user-friendly, command driven, computer-aided-control-system-design (CACSD) package.

Polynomial Diophantine equations can be used to solve pole-placement problems and linear quadratic Gaussian (LQG) optimal control problems, for both analogue and digital single-input-single-output (SISO) systems. A description is presented of the how these algorithms are implemented in Program CC, a user-friendly, command driven, computer-aided-control-system-design (CACSD) package.